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Sample records for rice mutants induced

  1. Early ripening mutants induced by colchicine in rice

    Institute of Scientific and Technical Information of China (English)

    CAIGuohai; YANWanchao; CAOXin

    1993-01-01

    In 1981-1983, the frequency and range of the mutants induced by colchicine were investigated in M2 and M3 of two indica-rice cultivars.Seedlings of M2 and M3 were treated with 0.05% colchicine solution at 4-5 leaf stage.

  2. Diversity and Stability Study on Rice Mutants Induced in Space Environment

    Institute of Scientific and Technical Information of China (English)

    Wei-Hong Lu; Xin-Zhu Wang; Qi Zheng; Shuang-Hong Guan; Ping Xin; Ye-Qing Sun

    2008-01-01

    To further study the characteristics of changes on the molecular level of rice mutants induced in space environment, we analyzed proteins in leaves and seeds of four rice mutants (two high-tillering and two low-tillering) in the 8th and 9th generations after a 15-day spaceflight, and compared with their ground controls by two-dimentional polyacrylamide gel electrophoresis and reverse phase liquid chromatography (RPLC). In addition, the albumin, globulin, prolamine, glutelin, and amylose of the mutant seeds were analyzed by RPLC and ultra-violet spectrometry. The results showed that the low-abundance proteins of leaves in the peak tillering stage are more likely to be induced compared with their corresponding controls. The albumin, globulin, and prolamine of the mutant seeds revealed changes when compared with their controls, and the characteristics of changes in different mutants were stably inherited in the 8th and 9th generations, suggesting that they can be used as biomarkers to identity the mutants induced by spaceflight. Moreover, two proteins (SSP9111 and SSP6302) were found to be expressed with high intensity (two-fold change) in different mutants, which were both correlated with photosystem according to mass spectrometry and database searching.

  3. Analysis of SSLP and Soluble Protein Contents in Leaves of Mutants Induced by High Pressure in Rice (Oryza sativa)

    Institute of Scientific and Technical Information of China (English)

    HE Xiu-ying; XU Shi-ping; LIAO Yao-ping; MAO Xing-xue; WENG Ke-nan; CHEN Zhao-ming; CHEN Yue-han; XIAO Wan-sheng

    2004-01-01

    Rice variety Yuexiangzhan and its mutants induced by high pressure were studied using microsatellite markers and soluble protein content analyses. Eleven of the 88 microsatellite primer pairs showed evident polymorphisms repeatedly, and the polymorphic frequencies were 3.4-11.3% between the mutants and Yuexiangzhan. The polymorphic markers were randomly located on chromosomes. The more similar the plant types of the mutants like their original variety, the less polymorphic loci were detected. In addition, there was variation in the soluble protein contents among the leaves of mutants,and the contents were significantly lower than those of the original variety.

  4. Changes in Chlorophyll Fluorescence of Rice Mutants Induced by High Hydrostatic Pressure

    Institute of Scientific and Technical Information of China (English)

    BAI Cheng-ke; LI Gui-shuang; PENG Chang-lian; DUAN Jun

    2003-01-01

    Three mutants of rice (Oryza sativa L. ), Mutant 1, Mutant 2and Mutant 3, which were selected by high hydrostatic pressure (75 MPa), and their parent Yuexiangzhan were used to study the changes in chlorophyll fluorescence during different growth stages. In all the three mutants, the function of PSⅡ was improved, Fy/Fm ratio of mutants increased compared to their parent at tillering and heading stage, and φPS Ⅱ also improved except for Mutant 2 at heading stage. Similar to their parent, the mutants exhibited slight photoinhibition at noon and almost complete recovery to initial levels of 6:00 after 18:00 at heading stage. At milking stage, the photoinhibition in the mutants was obvious, and recovered rapidly compared to the parent. Yields of individual plant and grain/straw ratio were also higher in three mutants than the parent. Results indicated that characteristics of chlorophyll fluorescence in leaves of mutants and their photoinhibition in the field had changed. It is suggested that high hydrostatic pressure induction could be applied as a new effective approach in high-yield rice breeding in the future.

  5. Alterations in DNA methylation and genome structure in two rice mutant lines induced by high pressure

    Institute of Scientific and Technical Information of China (English)

    SHEN; Sile; WANG; Zhenwei; SHAN; Xiaohui; WANG; Hua; LI; Ling; LIN; Xuyun; LONG; Likun; WENG; Kenan; LIU; Bao; ZOU; Guangtian

    2006-01-01

    By using high-pressure treatment, two mutant lines were obtained from a genetically stable japonica rice cultivar Bijing38. Genomic DNA of the mutant lines, together with the original line (Bijing38), was either undigested or digested by Hpa II/Msp I, and then subjected to molecular analysis using two markers, ISSR and RAPD. Results indicated that changes in the PCR amplification profiles of both markers are apparent in the two mutant lines compared with the original rice cultivar, suggesting that there had been both sequence changes and DNA methylation modifications in the mutant lines. Southern blot analysis using diverse sequences, including two cellular genes (S2 and S3), a set of retrotransposons (Osr7, Osr36, Tos19 and more), and a MITE transposon family (mPing and Pong), confirmed the results, and indicated that changes in DNA methylation pattern, genomic structure, and possible activation of some transposons indeed occurred in the mutant lines. Moreover, these changes are stably maintained through selfed generations and in different organs. Thus, our results indicate that it is possible to obtain stable mutants in rice by high pressure treatments, and the molecular basis of the mutants may include both genetic and epigenetic changes. Therefore, high hydrostatic pressure seems a promising approach for plant mutagenesis.

  6. Analysis of SSLP and Soluble Protein Contents in Leaves of Mutants Induced by High Pressure in Rice (Oryza sativa)

    Institute of Scientific and Technical Information of China (English)

    HEXiu-ying; XuShi-ping; LIAoYao-ping; MAOXing-xue; WENGKe-nan; CHENZhao-ming; CHENYue-han; XIAOWan-sheng

    2004-01-01

    Rice variety Yuexiangzhan and its mutants induced by high pressure were studied using microsatellite markers and soluble protein content analyses. Eleven of the 88 microsatellite primer pairs showed evident polymorphisms repeatedly, and the polymorphic frequencies wcrc 3.4-11.3% between the mutants and Yuexiangzhan. The polymorphic markers were randomly located on chromosomes. The more similar the plant types of the mutants like their original variety., the less polymorphic loci were detected. In addition, there was variation in the soluble protein contents among the leaves of mutants, and the contents were significantly lower than those of the original variety.

  7. Identification of a Long Rice Spikelet Mutant

    Institute of Scientific and Technical Information of China (English)

    WU Xian-jun; WANG Bin; HAN Zan-ping; XIE Zhao-hui; MOU Chun-hong; WANG Xu-dong

    2004-01-01

    A spontaneously occurring rice (Oryza sativa L. ) mutant, characterized by homeotic conversion in glumes and stamens, was found in the progeny of a cross. The mutant showed long glumes and glumaceous lodicules and morphological transformation of stamens into pistils. Mutant florets consisted of 1 to 3 completely developed pistils, some pistilloid stamens with filaments, but tipped by bulged tissue and 0 to 3 stigmas. It seens that the mutant phenotype of the homeotic conversions in glumes and stamens is similar to that of the B loss-of-function mutants in Arabidopsis and Antirrhinum. The mutant is controlled by a single recessive gene as a segregation ratio of 3:1 (wild type to mutant plants) was observed in the F2 generation.

  8. T-DNA插入产生的水稻小粒突变体的遗传分析%Genetic Analysis of a Small Grain Mutant Induced by T-DNA Insertion in Rice

    Institute of Scientific and Technical Information of China (English)

    张向前; 朱海涛; 邹金松; 曾瑞珍

    2008-01-01

    [Objective] The aim of this study is to understand the genetic characteristics of a grain shape mutant and its possible role in genetic improvement of grain yield in rice. [Method] On the basis of the collection of T-DNA tag lines, the progeny of homozygous plants carrying T-DNA insertion were screened for mutants with mutated phenotypes. The genetic analysis of the mutant and test for the linkage between the mutated phenotype and the T-DNA insertion were carried out to determine its genetic characteristics. [Result] In the present study, a grain shape mutant induced by T-DNA insertion in rice was identified, which showed small grain. Genetic analysis of the mutant showed that the two types of phenotype, normal and small grain in the segregating populations derived resistance showed that all the mutants were resistant while the normal results indicated that the mutant phenotype cosegregated with Bar gene. The small grain mutant caused by T-DNA insertion was confirmed by PCR amplification aiming at T-DNA. [Conclusion] The grain shape mutant is useful for isolation of the tagged gene and genetic improvement in rice.

  9. Expression of OsSPY and 14-3-3 genes involved in plant height variations of ion-beam-induced KDML 105 rice mutants

    Energy Technology Data Exchange (ETDEWEB)

    Phanchaisri, Boonrak [Science and Technology Research Institute, Chiang Mai University, Chiang Mai 50200 (Thailand); Molecular Biology Laboratory, Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai 50200 (Thailand); Samsang, Nuananong [Molecular Biology Laboratory, Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai 50200 (Thailand); Yu, Liang Deng; Singkarat, Somsorn [Plasma and Beam Physics Research Facility, Department of Physics and Materials Science, Faculty of Science, Chiang Mai University, Chiang Mai 50200 (Thailand); Thailand Center of Excellence in Physics, Commission on Higher Education, 328 Si Ayutthaya Road, Bangkok 10400 (Thailand); Anuntalabhochai, Somboon, E-mail: soanu.1@gmail.com [Molecular Biology Laboratory, Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai 50200 (Thailand)

    2012-06-01

    The culm length of two semidwarf rice mutants (PKOS1, HyKOS1) obtained from low-energy N-ion beam bombardments of dehusked Thai jasmine rice (Oryza sativa L. cv. KDML 105) seeds showed 25.7% and 21.5% height reductions and one spindly rice mutant (TKOS4) showed 21.4% increase in comparison with that of the KDML 105 control. A cDNA-RAPD analysis identified differential gene expression in internode tissues of the rice mutants. Two genes identified from the cDNA-RAPD were OsSPY and 14-3-3, possibly associated with stem height variations of the semidwarf and spindly mutants, respectively. The OsSPY gene encoded the SPY protein which is considered to be a negative regulator of gibberellin (GA). On the other hand, the 14-3-3 encoded a signaling protein which can bind and prevent the RSG (repression of shoot growth) protein function as a transcriptional repressor of the kaurene oxidase (KO) gene in the GA biosynthetic pathway. Expression analysis of OsSPY, 14-3-3, RSG, KO, and SLR1 was confirmed in rice internode tissues during the reproductive stage of the plants by semi-quantitative RT-PCR technique. The expression analysis showed a clear increase of the levels of OsSPY transcripts in PKOS1 and HyKOS1 tissue samples compared to that of the KDML 105 and TKOS4 samples at the age of 50-60 days which were at the ages of internode elongation. The 14-3-3 expression had the highest increase in the TKOS4 samples compared to those in KDML 105, PKOS1 and HyKOS1 samples. The expression analysis of RSG and KO showed an increase in TKOS4 samples compared to that of the KDML 105 and that of the two semidwarf mutants. These results indicate that changes of OsSPY and 14-3-3 expression could affect internode elongation and cause the phenotypic changes of semidwarf and spindly rice mutants, respectively.

  10. Status and Perspectives on the Researches of Rice Glutelin Mutants

    Institute of Scientific and Technical Information of China (English)

    ZHENG Tian-qing; SHEN Wen-biao; ZHU Su-song; ZHAI Hu-qu; WAN Jian-min

    2003-01-01

    Rice (Oryza sativa L. ) is one of the model plants for genomics research. As the raising offunctional rice breeding for special usage, glutelin mutants play a more and more important role in the func-tional rice breeding as well as eukaryotic gene expression and regulation research materials. For example, therice cultivar special for the patients suffering from kidney disease and diabetes could be developed from the riceglutelin mutants. In this paper, current researches on characterization, mutation mechanism and breeding us-age of various rice glutelin mutants, especially the low glutelin content cultivars, were all discussed with per-spectives on the trends of the glutelin mutant researches in the era of post-genomics.

  11. A wilty mutant of rice has impaired hydraulic conductance.

    Science.gov (United States)

    Koizumi, Koji; Ookawa, Taiichiro; Satoh, Hikaru; Hirasawa, Tadashi

    2007-08-01

    The rice CM2088 mutant is the wilty phenotype and wilts markedly under well-watered sunny conditions. The leaf water potential and epidermal (mainly stomatal) conductance of CM2088 plants decreased significantly under conditions that induced intense transpiration, as compared with those of wild-type plants, revealing that the wilty phenotype was not the result of abnormal stomatal behavior but was due to an increase in resistance to water transport. The resistance to water transport was dramatically elevated in the node and the sheath and blade of a leaf of the mutant, but not in the root or stem. The diameter of xylem vessels in the large vascular bundles of the leaf sheath and the internode tended to be small, and the numbers of vessel elements with narrowed or scalariform perforation plates in the leaf blade and sheath were greater in the mutant than in the wild type. Most xylem vessels were occluded, with air bubbles in the leaf sheath of the mutant during the midday hours under intense transpiration conditions, while no bubbles were observed in plants that were barely transpiring, revealing that the significant increase in resistance to water transport was a result of the cavitation. The additive effects of cavitation in xylem vessels and the decreased diameter and deformed plates of vessel elements might be responsible for the wilty phenotype of CM2088.

  12. Rice mutant resources for gene discovery

    NARCIS (Netherlands)

    Hirochika, H.; Guiderdoni, E.; An, G.; Hsing, Y.I.; Eun, M.Y.; Han, C.D.; Upadhyaya, N.; Ramachandran, S.; Zhang, Q.F.; Pereira, A.B.; Sundaresan, V.; Leung, H.

    2004-01-01

    With the completion of genomic sequencing of rice, rice has been firmly established as a model organism for both basic and applied research. The next challenge is to uncover the functions of genes predicted by sequence analysis. Considering the amount of effort and the diversity of disciplines requi

  13. Preliminary study on a gravity-insensitive rice mutant

    Institute of Scientific and Technical Information of China (English)

    金静; 朱诚; 张红心; 孙宗修

    2004-01-01

    A gravity-insensitive mutant was isolated from rice (Oryza sativa L. Cv. Zhonghua 11) transformed by Agrobacterium tumefaciens. The mutant's shoot growth (prostrate growth) was insensitive to gravity; whereas root growth displayed a normal positive gravitropism.Histological observation of root caps and leaf sheaths indicated that there was no significant difference in the number and size of amyloplasts in cells of the mutant and cells of the wild type

  14. Preliminary study on a gravity-insensitive rice mutant

    Institute of Scientific and Technical Information of China (English)

    金静; 朱诚; 张红心; 孙宗修

    2004-01-01

    A gravity-insensitive mutant was isolated from rice (Oryza sativa L. cv. Zhonghua 11) transformed by Agrobacterium tumefaciens. The mutant's shoot growth (prostrate growth) was insensitive to gravity; whereas root growth displayed a normal positive gravitropism. Histological observation of root caps and leaf sheaths indicated that there was no significant difference in the number and size of amyloplasts in cells of the mutant and cells of the wild type.

  15. The use of induced mutation combined with crossing in high quality rice breeding

    Energy Technology Data Exchange (ETDEWEB)

    Do Huu At; Bui Huy Thuy; Nguyen Van Bich; Tran Duy Quy [Agricultural Genetics Institute, Division of Genetics and Hybrid Rice Technology, Hanoi (Viet Nam); Nguyen Minh Cong [Hanoi No. 1 Teacher Training Univ., Department of Genetics (Viet Nam)

    2001-03-01

    The high quality rice varieties: Tam thom mutant rice Var., DT17 rice Var, DT21 glutinous rice Var were formed by induced mutation combined with crossing. Tam thom mutant rice Var. lost photosensitivity, could be planted 2 crops/year. DT17 rice Var with high yielding capacity, suitable for growth on lowland in summer crop, is replacing step-by-step Moctuyen rice Var. in North Vietnam. DT21 glutinous rice Var. could be planted 2 crops/year and had short growth duration, average yield was 4.0-4.5 tons/ha. These three ones had good quality, soft and scent cooked rice, suitable for customers and export requirements. Tam thom mutant rice Var. DT17 rice Var., DT21 and glutinous rice Var. were adopted for regional production by Ministry of Agriculture and Rural Development and allowed to be in trial production. (author)

  16. A Rapid DNA Mini-prep Method for Large-Scale Rice Mutant Screening

    Institute of Scientific and Technical Information of China (English)

    QIU Fu-lin; WANG He-he; CHEN Jie; ZHUANG Jie-yun; Hei LEUNG; CHENG Shi-hua; Wu Jian-li

    2006-01-01

    A high throughput rice DNA mini-preparation method was developed. The method is suitable for large-scale mutant bank screening as well as large mapping populations with characteristics of maintaining relatively high level of DNA purity and concentration. The extracted DNA was tested and suitable for regular PCR amplification (SSR) and for Targeting Induced Local Lesion in Genome (TILLING) analysis.

  17. Morphology and inheritance of a new rice leaf death mutant

    Institute of Scientific and Technical Information of China (English)

    CHENGZhakuan; GUMingkang

    1998-01-01

    A new mutant showing successive leaf death was selected in a mutmion populalion from an indica rice variety Zhongxian 3037 trealed with 94Co-γ rays. The leaves of this rnutanl began lodie on the emergence of tlle third leaf tip, and all leaves died sooner alter flowering. Consequently,

  18. A dominant semi dwarf mutant in rice

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    @@ In the winter of 1997, a semi dwarf mutant was found in the F6 population of M9056/ R8018 xuan in Hainan Province. In the spring of 1998, the seeds were sown in Hefei, Anhui Province and the plant height of the population was measured at maturity.

  19. Phenotypic Characterization of a Female Sterile Mutant in Rice

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    A female sterile mutant, derived from a spontaneous mutation, wasfirst discovered in rice (Oryza sativa L. ssp.indica) restorer line 202R. With normal flowering, the mutant exhibits an extremely Iow seed-setting rate. When the mutant is crossed as a pollen donor, the seeds set normally; whereas when it is used as a pollen receiver,no seeds are obtained even with mixed pollen grains of different varieties sprinkled over the stigmas. The floret of the mutant, consisting of six stamens and one pistil, looks the same as that of the wild type in the malefemale organs, except that less than 10% of the mutant florets have three stigmas on the ovary. Although the mutant has a low seed-setting rate, Its pollen fertility is approximately 87.1%, which is equal to that of the wild type. In addition, more than 90% of the mature embryo sacs of the mutant have complete inner structures. At every stage after pollination, the sperm, embryo, and endosperm are not found in the mutant embryo sac,whereas the disintegration of the egg cell that does not accomplish fertilization is visible. Through observations with a fluorescence microscope, we have found that the pollen grains germinate normally, whereas the pollen tube abnormally elongates in the style-transmitting tissue. The mutant pollen tubes display various defects in the style, such as slower elongation, conversed elongation, distorted elongation, swollen tips, or branched tips. As a result, the growth of the pollen tubes ceases in the style, and, therefore, the pollen tubes cannot reach the embryo sac and the process of double fertilization is blocked. Based on these observations,we conclude that this mutant, designated as fs-202R, is a novel type of female sterile mutation in rice, which causes the arrest of the elongation of the pollen tube.

  20. Morphological Structure and Genetic Mapping of New Leaf-Color Mutant Gene in Rice (Oryza sativa)

    Institute of Scientific and Technical Information of China (English)

    LI Yu-hong; WANG Bao-he; DAI Zheng-yuan; LI Ai-hong; LIU Guang-qing; ZUO Shi-min; ZHANG Hong-xi; PAN Xue-biao

    2012-01-01

    Leaf-color mutations are a widely-observed class of mutations,playing an important role in the study of chlorophyll biosynthesis and plant chloroplast structure,function,genetics and development.A naturally-occurring leaf-color rice mutant,Baihuaidao 7,was analyzed.Mutant plants typically exhibited a green-white-green leaf-color progression,but this phenotype was only expressed in the presence of a stress signal induced by mechanical scarification such as transplantation.Prior to the appearance of white leaves,mutant plant growth,leaf color,chlorophyll content,and chloroplast ultrastructure appeared to be identical to those of the wild type.After the changeover to white leaf color,an examination of the mutated leaves revealed a decrease in total chlorophyll,chlorophyll a,chlorophyll b,and carotenoid content,a reduction in the number of chloroplast grana lamella and grana,and a gradual degradation of the thylakoid lamellas.At maturity,the mutant plant was etiolated and dwarfed compared with wild-type plants.Genetic analysis indicated that the leaf mutant character is controlled by a recessive nuclear gene.Genetic mapping of the mutant gene was performed using an F2 population derived from a Baihuaidao 7 ×Jiangxi 1587 cross.The mutant gene was mapped to rice chromosome 11,positioned between InDel markers L59.2-7 and L64.8-11,which are separated by approximately 740.5 kb.The mutant gene is believed to be a new leaf-color mutant gene in rice,and is tentatively designated as gwgl.

  1. Differential gene expression in salt-tolerant rice mutant and its parental variety

    Institute of Scientific and Technical Information of China (English)

    张劲松; 周骏马; 张驰; 陈受宜

    1996-01-01

    The differential expressions of three genes rbcL, salT and rab!6 in response to ABA, NaCl, PEG and heat shock were investigated in seedlings of a salt-tolerant rice mutant 20 (mutant 20) and its parental variety Oryza sativa var. japonica 77-170(170). By Northern blot analysis it was found that ABA induced the expression of all three genes of rbcL, salT and rab16 in shoots and roots of both 170 and mutant 20 with the exceptions of rab16 in shoots of mutant 20 and rbcL in roots of 170. Lower concentrations of NaCl induced rbcL expression in shoots of mutant 20 but not 170. Higher concentrations of NaCl decreased rbcL expression but induced expressions of salT and rab16 in shoots of both 170 and mutant 20. PEG(15%) and 37℃ heat shock showed almost no effects on the expression of the three genes in mutant 20. However, they caused a decrease in rbcL expression and slight induction of the rab16 gene in 170, with salT expression unaffected. These results indicated that mutant 20 was relatively less responsiv

  2. Metabolite profiling of two low phytic acid (lpa) rice mutants.

    Science.gov (United States)

    Frank, Thomas; Meuleye, Bertrand Seumo; Miller, Andreas; Shu, Qing-Yao; Engel, Karl-Heinz

    2007-12-26

    Two low phytic acid (lpa) rice mutant lines, Os-lpa-XS110-1 and Os-lpa-XS110-2, were grown together with their parent wild-type variety Xiushui 110 in four field trials. HPLC analysis of inositol phosphates in the seeds produced demonstrated that compared to the wild-type, the reduction in phytic acid content in Os-lpa-XS110-1 (-46%) was more pronounced than that in Os-lpa-XS110-2 (-23%). Lower inositol phosphates (InsP 3, InsP 4, InsP 5) were not detected in the mutants. The lpa mutants and the wild-type rice were subjected to comparative metabolite profiling by capillary gas chromatography. On average, 34% (Os-lpa-XS110-1) and 42% (Os-lpa-XS110-2) of the detected peaks were statistically significantly different between wild-type and mutants. However, only a few of these differences could be consistently observed for all field trials. Identification and quantification of the consistently different metabolites revealed that contents of myo-inositol and raffinose were increased in Os-lpa-XS110-1 but decreased in Os-lpa-XS110-2 compared to the wild-type. In addition, Os-lpa-XS110-1 exhibited increased levels of galactose and galactinol. Consideration of these metabolic changes in light of the routes involved in the biosynthesis of phytic acid indicated a disturbance in the early biosynthetic pathway of phytic acid in Os-lpa-XS110-2 (similar to the lpa-1 type mutation in maize) and a mutation event affecting phosphorylation of myo-inositol in Os-lpa-XS110-1 (similar to the lpa-3-type mutation).

  3. Identification of a Gravitropism-Deficient Mutant in Rice

    Directory of Open Access Journals (Sweden)

    He Yan

    2017-03-01

    Full Text Available A gravitropism-deficient mutant M96 was isolated from a mutant bank, generated by ethyl methane sulfonate (EMS mutagenesis of indica rice accession ZJ100. The mutant was characterized as prostrate growth at the beginning of germination, and the prostrate growth phenotype ran through the whole life duration. Tiller angle and tiller number of M96 increased significantly in comparison with the wild type. Tissue section observation analysis indicated that asymmetric stem growth around the second node occurred in M96. Genetic analysis and gene mapping showed that M96 was controlled by a single recessive nuclear gene, tentatively termed as gravitropism-deficient M96 (gdM96, which was mapped to a region of 506 kb flanked by markers RM5960 and InDel8 on the long arm of chromosome 11. Sequencing analysis of the open reading frames in this region revealed a nucleotide substitution from G to T in the third exon of LOC_Os11g29840. Additionally, real-time fluorescence quantitative PCR analysis showed that the expression level of LOC_Os11g29840 in the stems was much higher than in the roots and leaves in M96. Furthermore, the expression level was more than four times in M96 stem than in the wild type stem. Our results suggested that the mutant gene was likely a new allele to the reported gene LAZY1. Isolation of this new allele would facilitate the further characterization of LAZY1.

  4. Photosynthetic Characteristics of Flag Leaves in Rice White Stripe Mutant 6001 During Senescence Process

    Directory of Open Access Journals (Sweden)

    Xiao-hui ZHEN

    2014-11-01

    Full Text Available Physiological, biochemical and electron microscopy analyses were used to investigate the photosynthetic performance of flag leaves in rice white stripe mutant 6001 during the senescence process. Results showed that the chlorophyll content at the heading and milk-ripe stages in rice mutant 6001 were about 34.78% and 3.00% less than those in wild type 6028, respectively. However, the chlorophyll content at the fully-ripe stage in rice mutant 6001 was higher than that in wild type 6028. At the heading stage, the net photosynthetic rate (Pn in rice mutant 6001 was lower than that in wild type 6028. Rice mutant 6001 also exhibited a significantly slower decrease rate of Pn than wild type 6028 during the senescence progress, especially at the later stage. Furthermore, Ca2+-ATPase, Mg2+-ATPase and photophosphorylation activities exhibited the similar trends as the Pn. During the senescence process, the 68 kDa polypeptide concentrations in the thylakoid membrane proteins exhibited a significant change, which was one of the critical factors that contributed to the observed change in photosynthesis. We also observed that the chloroplasts of rice mutant 6001 exhibited higher integrity than those of wild type 6028, and the chloroplast membrane of rice mutant 6001 disintegrated more slow during the senescence process. In general, rice mutant 6001 had a relatively slower senescence rate than wild type 6028, and exhibited anti-senescence properties.

  5. Agronomic traits and RAPD analysis of two mutants derived from rice somatic cell culturing

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Genetic variation, including agronomic trait variation, often occurs in somatic cell culturing. In this study, we compared the main agronomic traits of two rice mutants, M3 and M14, which were derived from Shenxiangjing 5 somatic cell culturing. Significant differences were found between the two mutants and the wild rice Shenxiangjing 5 (Table 1). Results were as follows:

  6. Photosynthetic Characteristics of Flag Leaves in Rice White Stripe Mutant 6001 During Senescence Process

    Institute of Scientific and Technical Information of China (English)

    ZHEN Xiao-hui; XU Jin-gang; SHEN Wei-jun; ZHANG Xiao-juan; ZHANG Qi-jun; LU Chuan-gen; CHEN Guo-xiang; GAO Zhi-ping

    2014-01-01

    Physiological, biochemical and electron microscopy analyses were used to investigate the photosynthetic performance of flag leaves in rice white stripe mutant 6001 during the senescence process. Results showed that the chlorophyll content at the heading and milk-ripe stages in rice mutant 6001 were about 34.78% and 3.00% less than those in wild type 6028, respectively. However, the chlorophyll content at the fully-ripe stage in rice mutant 6001 was higher than that in wild type 6028. At the heading stage, the net photosynthetic rate (Pn) in rice mutant 6001 was lower than that in wild type 6028. Rice mutant 6001 also exhibited a significantly slower decrease rate ofPn than wild type 6028 during the senescence progress, especially at the later stage. Furthermore, Ca2+-ATPase, Mg2+-ATPase and photophosphorylation activities exhibited the similar trends as thePn. During the senescence process, the 68 kDa polypeptide concentrations in the thylakoid membrane proteins exhibited a significant change, which was one of the critical factors that contributed to the observed change in photosynthesis. We also observed that the chloroplasts of rice mutant 6001 exhibited higher integrity than those of wild type 6028, and the chloroplast membrane of rice mutant 6001 disintegrated more slow during the senescence process. In general, rice mutant 6001 had a relatively slower senescence rate than wild type 6028, and exhibited anti-senescence properties.

  7. Characterization and Fine Mapping of the ibf Mutant in Rice

    Institute of Scientific and Technical Information of China (English)

    Jiajun Cui; Shengci Fan; Tian Shao; Zejun Huang; Dali Zheng; Ding Tang; Ming Li; Qian Qian; Zhukuan Cheng

    2007-01-01

    The pigment is an important character in plant development. In the present study, we characterized and fine mapped one inhibitor for brown furrows gene (ibf) in rice (Oryza sativa L.). in the Ibf mutant, brown pigments specifically accumulate in the furrows of hulls as seeds mature and reach a maximum level in dry seeds. Genetic analysis showed that the mutant phenotype is controlled by one recessive nuclear gene, which was finally mapped in a 90-kb reglon on the long arm of chromosome 9. Polymerase chain reaction and Southern blotting analysis revealed that there was a 26 kb deletion in the 90-kb region in the mutant. Since all the open reading frames outside the gap in the delimited reglon had no detectable difference in DNA sequence with the wild-type, we postulated that the Ibf locus should be located in the gap. Through gene annotation and reverse transcription-polymerase chain reaction (RT-PCR) analysis, we selected OsKF1 encoding a kelch repeat-containing F-box family protein as the candidate gene of ibf.

  8. Genetic Analysis and Gene Mapping of Light Brown Spotted Leaf Mutant in Rice

    Institute of Scientific and Technical Information of China (English)

    FENG Bao-hua; YANG Yang; SHI Yong-feng; LIN Lu; CHEN Jie; WEI Yan-lin; Hei LEUNG

    2013-01-01

    A light brown spotted-leaf mutant of rice was isolated from an ethane methyl sulfonate (EMS)induced IR64 mutant bank.The mutant,designated as Ibsl1 (light brown spotted-leaf 1),displayed light brown spot in the whole growth period from the first leaf to the flag leaf under natural summer field conditions.Agronomic traits including plant height,growth duration,number of filled grains per panicle,seed-setting rate and 1000-grain weight of the mutant were significantly affected.Genetic analysis showed that the mutation was controlled by a single recessive gene,tentatively named Ibsl1(t),which was mapped to the short arm of chromosome 6.By developing simple sequence repeat (SSR) markers,the gene was finally delimited to an interval of 130 kb between markers RM586 and RM588.The Ibsl1(t) gene is likely a novel rice spotted-leaf gene since no other similar genes have been identified near the chromosomal region.The genetic data and recombination populations provided will facilitate further fine-mapping and cloning of the gene.

  9. Uptake,Subcellular Distribution,and Chemical Forms of Cadmium in Wild-Type and Mutant Rice

    Institute of Scientific and Technical Information of China (English)

    HE Jun-Yu; ZHU Cheng; REN Yan-Fang; YAN Yu-Ping; CHENG Chang; JIANG De-An; SUN Zong-Xiu

    2008-01-01

    Wild-type (Zhonghua 11) and mutant rice (Oryza sativa L.) plants were used to investigate the effect of cadmium (Cd) application on biomass production,to characterize the influx of Cd from roots to shoots,and to determine the form,content,and subcellular distribution of Cd in the roots,leaf sheaths,and leaves of the rice plants.Seedlings were cultivated in a nutrient solution and were treated with 0.5 mmol L-1 of Cd2+ for 14 d.The sensitivity of rice plants to Cd toxicity was tested by studying the changes in biomass production and by observing the onset of toxicity symptoms in the plants.Both the wild-type and mutant rice plants developed symptoms of Cd stress.In addition,Cd application significantly (P ≤ 0.01) decreased dry matter production of roots,leaf sheaths,and leaves of both types,especially the mutant.The Cd content in roots of the mutant was significantly (P≤0.05) higher than that of the wild-type rice.However,there was no significant difference in the Cd content of roots,leaf sheaths,and leaves between the wild-type and mutant rice.Most of the Cd was bound to the cell wall of the roots,leaf sheaths,and leaves,and the mutant had greater Cd content in cell organelles than the wild type.The uneven subcellular distribution could be responsible for the Cd sensitivity of the mutant rice.Furthermore,different chemical forms of Cd were found to occur in the roots,leaf sheaths,and leaves of both types of rice plants.Ethanol-,water-,and NaCl-extractable Cd had greater toxicity than the other forms of Cd and induced stunted growth and chlorosls in the plants.The high Cd content of the toxic forms of Cd in the cell organelles could seriously damage the cells and the metabolic processes in mutant rice plants.

  10. Characterization and mapping of a novel light-dependent lesion mimic mutant lmm6 in rice (Oryza sativa L.)

    Institute of Scientific and Technical Information of China (English)

    XIAO Gui-qing; ZHANG Hai-wen; LU Xiang-yang; HUANG Rong-feng

    2015-01-01

    A novel rice lesion mimic mutant (LMM) was isolated from an ethane methyl sulfonate (EMS)-induced 02428 mutant bank. The mutant, tentatively designated as lmm6, develops necrotic lesions in the whole growth period along with changes in several important agronomic traits. We found that the initiation of the lesions was induced by light and cel death occurred in lmm6 accompanied with accumulation of reactive oxygen species (ROS). The lower chlorophyl content, soluble protein content and superoxide dismutase (SOD) activity, the higher malondialdehyde (MDA) content were detected in lmm6 than in the wild type (WT). Moreover, the observation by transmission electronic microscope (TEM) demonstrated that some organel es were damaged and the stroma lamel a of chloroplast was irregular and loose in mesophyl cel of lmm6. In addition, lmm6 was more resistant than WT to rice blast fungus Magnaporthe grisea infection, which was consistent with increased expression of four genes involved in the defense-related reaction. Genetic analysis showed that mutant trait of lmm6 is inherited as a monogenic recessive nuclear gene located on the long arm of chromosome 6. Using simple sequence repeat (SSR) markers, the target gene was ifnal y delimited to an interval of 80.8 kb between markers MM2359 and MM2370, containing 7 annotated genes. Taken together, our results provide the information to identify a new gene involved in rice lesion mimic, which wil be helpful in clarifying the mechanism of cel death and disease resistance in rice.

  11. Changes of carbohydrate and protein metabolism in seedling leaves of a temperature-induced greenable albino mutant line W25 of rice

    Institute of Scientific and Technical Information of China (English)

    WU; SHUQingyao; XIAYingwu

    1998-01-01

    W25 is a low-temperature sensitive albino mutant line, Temperature not only controls the albino phenotype expression of W2o, but also determines whether it could survive, When the temperature is lower than 25℃. the leaves of W25 shows complete albino, but they exhibits normal green when temperature is higher than 30℃

  12. Isolation and characterization of a low phytic acid rice mutant reveals a mutation in the rice orthologue of maize mik.

    Science.gov (United States)

    Using a forward genetics approach, we isolated two independent low phytic acid (lpa) rice mutants, N15-186 and N15-375. Both mutants are caused by single gene, recessive non-lethal mutations which result in approximately 75% (N15-186) and 43% (N15-375) reductions in seed phytic acid (inositol hexaki...

  13. PHENOTYPIC ANALYSIS OF OsTPKb LOSS OF FUNCTION MUTANT RICE LINES

    Directory of Open Access Journals (Sweden)

    Isayenkov S. V.

    2015-08-01

    Full Text Available The results of screen and analysis of two OsTPKb rice mutant lines were described. The phenotypes and growth rate level of homozygous mutant plants of both rice lines were estimated. The electron microscopy of aleurone layer from forming seeds was performed. The OsTPKb mutant plants demonstrate lower growth rate in comparison with wild type plants. The loss of function OsTPKb mutations invariably led to (semisterile rice plants. The functional disruption of OsTPKb channel has negative impact on plant growth and development. It might completely change the cell morphology of aleurone layer.

  14. Isolation and characterization of a spotted leaf 32 mutant with early leaf senescence and enhanced defense response in rice

    Science.gov (United States)

    Sun, Liting; Wang, Yihua; Liu, Ling-long; Wang, Chunming; Gan, Ting; Zhang, Zhengyao; Wang, Yunlong; Wang, Di; Niu, Mei; Long, Wuhua; Li, Xiaohui; Zheng, Ming; Jiang, Ling; Wan, Jianmin

    2017-01-01

    Leaf senescence is a complex biological process and defense responses play vital role for rice development, their molecular mechanisms, however, remain elusive in rice. We herein reported a rice mutant spotted leaf 32 (spl32) derived from a rice cultivar 9311 by radiation. The spl32 plants displayed early leaf senescence, identified by disintegration of chloroplasts as cellular evidence, dramatically decreased contents of chlorophyll, up-regulation of superoxide dismutase enzyme activity and malondialdehyde, as physiological characteristic, and both up-regulation of senescence-induced STAY GREEN gene and senescence-associated transcription factors, and down-regulation of photosynthesis-associated genes, as molecular indicators. Positional cloning revealed that SPL32 encodes a ferredoxin-dependent glutamate synthase (Fd-GOGAT). Compared to wild type, enzyme activity of GOGAT was significantly decreased, and free amino acid contents, particularly for glutamate and glutamine, were altered in spl32 leaves. Moreover, the mutant was subjected to uncontrolled oxidative stress due to over-produced reactive oxygen species and damaged scavenging pathways, in accordance with decreased photorespiration rate. Besides, the mutant showed higher resistance to Xanthomonas oryzae pv. Oryzae than its wild type, coupled with up-regulation of four pathogenesis-related marker genes. Taken together, our results highlight Fd-GOGAT is associated with the regulation of leaf senescence and defense responses in rice. PMID:28139777

  15. Induced resistance in rice against insects.

    Science.gov (United States)

    Karban, R; Chen, Y

    2007-08-01

    Vaccinations are the mainstay of western preventive medicine, and they have been used to protect some crops against disease and insect pests. We consider rice as a model for protection using induced resistance since it is one of the most important staple crops and there have been significant new developments in: cross-resistance among rice insects, chemical pathways involved in induced resistance, sequencing the rice genome and expression of genes conferring resistance against rice insect pests. Insect attack has been found to cause lesions that kill planthopper eggs and early stages of gall midges. Damaged plants released volatiles that made them less likely to be chosen by planthoppers and more attractive to parasitoids. Chemical elicitors have been developed for dicotyledonous plants and these can induce resistance in rice, although rice does not fit models developed to explain signalling in dicots. For example, salicylic acid did not increase in rice after infection by pathogens and did not appear to be the mobile signal for induced resistance against pathogens although it was involved in induced responses to phloem-feeding insects. Jasmonic acid acted as a signal in some induced responses to pathogens as well as chewing insects. Many of the genes associated with induced resistance in rice have recently been mapped, and techniques are being developed to incorporate them into the genome of cultivated varieties. Attempts to control insect pests of rice will affect interactions with pathogens, predators and parasites, and other organisms in this agroecosystem.

  16. Molecular mapping of split rice spikelet mutant srs-1 and analysis of its homeotic function in rice

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    srs-1, a new floral organ identity gene in rice, was mapped using RAPD and RFLP markers. Firstly, the cross was made between "ZhaiYeQing 8" (ZYQ8, indica) and split rice spikelet (SRS, japonica) mutant. The ratio of wild-type individuals and mutant plants in F2 population is 3:1, which indicates that the mutant characteristics are controlled by single recessive gene, srs-1. Consequently, BSA method was adopted and 520 random 10-mer primers were used to screen polymorphic bands between two bulks. Six primers could amplify polymorphic bands, of which S465 generates the most stable RAPD patterns. Then, S465 that cosegregates in F2 population has been converted into an RFLP marker successfully. Furthermore, srs-1 gene was mapped on chromosome 3 using DH mapping population. The effect of srs-1 gene results in the mutant of split rice spikelet. The mutant has longer and softer palea/lemma than those of wild-type, and two small palea/lemma-like organs between palea and lemma. In addition, there is a flower with three stamens and carpel in the axil of lemma. Thus, there are nine stamens and two carpels in the spikelet of mutant. srs-1 gene may belong to homeotic gene of class A according to the mutant characteristics and ABC model.

  17. Isolation and characterization of rice lesion mimic mutants from a T-DNA tagged population

    Institute of Scientific and Technical Information of China (English)

    LI Shutian; PEI Zhongyou; LUO Lijuan; TIAN Yingchuan; HE Chaozu

    2005-01-01

    A rice ( Oryza sativa L. ssp. japonica cv. Nipponbare) T-DNA tagged population consisting of about 7000 individual lines was generated and screened for rice lesion mimic mutants in the T1 generation. Ten lines were found to develop spontaneous lesions in the absence of pathogen infection and displayed distinct lesion phenotypes. These mutants were tentatively designated as lm1 -lm10 (for lesion mimic), respectively. Lesion formation of lm mutants was developmentally regulated, and all the mutants showed stunted growth and reduced fertility. Genetic analysis demonstrated that all the mutations were recessive, and five partially fertile mutants (lm4-lm8) were derived from different loci. Mimic lesions occurring on the leaves of lm mutants resulted from cell death as revealed by trypan blue staining. Six of them ( lm3 -lm8 ) exhibited enhanced resistance to five bacterial blight isolates, indicating their wide-spectrum resistance to this pathogen. These results imply that some lesion mimic mutations of rice might be involved in disease resistance signaling pathways,and that isolation of these mutated genes may be useful for elucidating molecular mechanisms of plant disease resistance. Among the mutants, only one mutant, lm6, was preliminarily shown to cosegregate with the inserted T-DNA in its T1 generation, making it feasible to isolate the gene responsible for the phenotype of this mutant.

  18. Molecular mapping of split rice spikelet mutant srs-1 and analysis of its homeotic function in rice

    Institute of Scientific and Technical Information of China (English)

    白素兰; 刘永胜; 孙敬三; 朱立煌

    2000-01-01

    srs-1, a new floral organ identity gene in rice, was mapped using RAPD and RFLP markers. Firstly, the cross was made between "ZhaiYeQing 8" (ZYQ8, indica) and split rice spikelet (SRS, japonica) mutant. The ratio of wild-type individuals and mutant plants in F2 population is 3:1, which indicates that the mutant characteristics are controlled by single recessive gene, srs-1. Consequently, BSA method was adopted and 520 random 10-mer primers were used to screen polymorphic bands between two bulks. Six primers could amplify polymorphic bands, of which S465 generates the most stable RAPD patterns. Then, S465 that cosegregates in F2 population has been converted into an RFLP marker successfully. Furthermore, srs-1 gene was mapped on chromosome 3 using DH mapping population. The effect of srs-1 gene results in the mutant of split rice spikelet. The mutant has longer and softer palea/lemma than those of wild-type, and two small palea/lemma-like organs between palea and lemma. In addition, there is a flower wit

  19. Characterization and mapping of a spotted leaf mutant in rice (Oryza sativa

    Directory of Open Access Journals (Sweden)

    Xue Xu

    2014-06-01

    Full Text Available Spotted leaf mutant belongs to a class of mutants that can produce necrotic lesions spontaneously in plants without any attack by pathogens. These mutants have no beneficial effect on plant productivity but provide a unique opportunity to study programmed cell death in plant defense responses. A novel rice spotted leaf mutant (spl30 was isolated through low-energy heavy ion irradiation. Lesion expression was sensitive to light and humidity. The spl30 mutant caused a decrease in chlorophyll and soluble protein content, with marked accumulation of reactive oxygen species (ROS around the lesions. In addition, the spl30 mutant significantly enhanced resistance to rice bacterial blight (X. oryzae pv. oryzae from China (C1-C7. The use of SSR markers showed that the spl30 gene was located between markers XSN2 and XSN4. The genetic distance between the spl30 gene and XSN2 and between spl30 and XSN4 was 1.7 cM and 0.2 cM, respectively. The spl30 gene is a new gene involved in lesion production and may be related to programmed cell death in rice. The ability of this mutant to confer broad resistance to bacterial blight provides a model for studying the interaction between plants and pathogenic bacteria.

  20. Characterization and Genetic Analysis of a Novel Mutant mst of Rice Defective in Flower Development

    Institute of Scientific and Technical Information of China (English)

    LI Yun; XU Pei-zhou; ZHANG Hong-yu; FU Shao-hong; YANG Jin; ZHANG Ru-quan; WU Xian-jun

    2009-01-01

    A spontaneous mutant with multiple stigmas (mst) was found in an indica rice line 466. The mst mutant exhibits normal at the vegetative development stage and produces normal inflorescence structures. The difference between the mutant and the wild type was observed when the stamen primordium began to develop. In the mst florets, palea and lemma opened, lodicules were homeotically transformed into palea/lemma-like structures, and stamens were homeotically transformed into carpel-like structures. It looked like multiple stigmas being full of the whole floret. The phenotypic changes of mst were very similar to that of B-like mutant spw1. Compared with other mutants with pistillate morphologies, the severe mst florets showed that the inner three floral organs were completely changed into palea/lemma-like structures. Moreover, the mutant was female sterile. Occasionally, with the changing environment, one or two stamens were fertile. Genetic analysis indicated that the mutant traits were controlled by a single recessive gene.

  1. Transcript profiling of crown rootless1 mutant stem base reveals new elements associated with crown root development in rice

    Directory of Open Access Journals (Sweden)

    Van Anh Le Thi

    2011-08-01

    Full Text Available Abstract Background In rice, the major part of the post-embryonic root system is made of stem-derived roots named crown roots (CR. Among the few characterized rice mutants affected in root development, crown rootless1 mutant is unable to initiate crown root primordia. CROWN ROOTLESS1 (CRL1 is induced by auxin and encodes an AS2/LOB-domain transcription factor that acts upstream of the gene regulatory network controlling CR development. Results To identify genes involved in CR development, we compared global gene expression profile in stem bases of crl1 mutant and wild-type (WT plants. Our analysis revealed that 250 and 236 genes are down- and up-regulated respectively in the crl1 mutant. Auxin induces CRL1 expression and consequently it is expected that auxin also alters the expression of genes that are early regulated by CRL1. To identify genes under the early control of CRL1, we monitored the expression kinetics of a selected subset of genes, mainly chosen among those exhibiting differential expression, in crl1 and WT following exogenous auxin treatment. This analysis revealed that most of these genes, mainly related to hormone, water and nutrient, development and homeostasis, were likely not regulated directly by CRL1. We hypothesized that the differential expression for these genes observed in the crl1 mutant is likely a consequence of the absence of CR formation. Otherwise, three CRL1-dependent auxin-responsive genes: FSM (FLATENNED SHOOT MERISTEM/FAS1 (FASCIATA1, GTE4 (GENERAL TRANSCRIPTION FACTOR GROUP E4 and MAP (MICROTUBULE-ASSOCIATED PROTEIN were identified. FSM/FAS1 and GTE4 are known in rice and Arabidopsis to be involved in the maintenance of root meristem through chromatin remodelling and cell cycle regulation respectively. Conclusion Our data showed that the differential regulation of most genes in crl1 versus WT may be an indirect consequence of CRL1 inactivation resulting from the absence of CR in the crl1 mutant. Nevertheless

  2. Agrobacterium mediated construction of rice mutant population with transposon

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Transposon tagging is a powerful tool for gene study, especially for genes among different plants, such as Arabidopsis, tobacco, and tomato, etc. This effective strategy for gene isolation has been applied to rice and progress has been made in recent years.

  3. Cadmium Accumulation and Its Toxicity in Brittle Culm 1 (bc1, a Fragile Rice Mutant

    Directory of Open Access Journals (Sweden)

    Guo-sheng SHAO

    2007-09-01

    Full Text Available Cadmium (Cd accumulation and toxicity in rice plants were characterized and identified by using brittle culm 1 (bc1, a fragile rice mutant and its wild type (Shuangkezao, an indica rice as materials by hydroponics. The low Cd level didn't obviously affect the growth parameters in both rice genotypes, but under high Cd levels (1.0 and 5.0 μmol/L, the growth of both rice plants were substantially inhibited. Moreover, bc1 tended to suffer more seriously from Cd toxicity than Shuangkezao. Cd accumulation in both rice plants increased with the increase of Cd levels. There was a significant difference in Cd accumulation between the two rice genotypes with constantly higher Cd concentration in bc1, which also accumulated more Cd at 0, 0.1, and 1.0 μmol/L Cd levels. The same case was found in the two rice plants grown on Cd-contaminated soil. This suggested that cell wall might play an important role in Cd accumulation in rice plants by the physiological mechanisms. The malondialdehyde (MDA content, superoxide dismutase (SOD and peroxidase (POD activities in rice plants were affected differently under Cd treatments, and which implied that POD might play the main role in detoxifying active oxygen free radical. A significant difference in antioxidative system between the two rice genotypes was found with constantly higher MDA content, SOD and POD activities in bc1. In summary, bc1 accumulated more Cd and appeared to be more sensitive to Cd stress compared with its wild type.

  4. Genetic Analysis and Molecular Mapping of a Novel Chlorophyll-Deficit Mutant Gene in Rice

    Institute of Scientific and Technical Information of China (English)

    HUANG Xiao-qun; WANG Ping-rong; ZHAO Hai-xin; DENG Xiao-jian

    2008-01-01

    A rice etiolation mutant 824ys featured with chlorophyll deficiency was identified from a normal green rice variety 824B.It showed whole green-yellow plant from the seedling stage,reduced number of tillers and longer growth duration.The contents of chlorophyll,chlorophyll a,chlorophyll b and net photosynthetic rate in leaves of the mutant obviously decreased,as well as the number of spikelets per panicle,seed setting rate and 1000-grain weight compared with its wild-type parent.Genetic analyses on F1 and F2 generetions of 824ys crossed with three normal green varieties showed that the chlorophyll-deficit mutant character was controlled by a pair of recessive nuclear gene.Genetic mapping of the mutant gene was conducted by using microsatellite markers and F2 mapping population of 495R/824ys,and the mutant gene of 824ys was mapped on the shon arm of rice chromosome 3.The genetic distances from the target gene to the markers RM218,RM282 and RM6959 were 25.6 cM,5.2 cM and 21.8 cM,respectively.It was considered to be a now chlorophyll-deficit mutant gene and tentatively named as chl11(t).

  5. Effect of Low Content Chlorophyll on Distribution Properties of Absorbed Light Energy in Leaves of Mutant Rice

    Institute of Scientific and Technical Information of China (English)

    XU Xiao-ming; ZHANG Rong-xian; TANG Yun-lai

    2004-01-01

    This paper reported the diurnal variations of photochemical efficiency of PSII, thermal dissipation rate and other physiology process in thelow content chlorophyll mutant rice and its wild type under field conditions, and analyzed the difference of absorbed light distribution between the two rice varieties in a day. The results showed that the mutant had poor absorbed light because of its little light absorption coefficient, but higher electron transportg rate could partly reduce the disadvantageous effect of deficient absorbed light in mutant. Compared with wild-type rice, the mutant had less excess excitation energy and the fraction of absorbed light allocated to photochemical process was more.

  6. Specific-Locus Amplified Fragment Sequencing Reveals Spontaneous Single-Nucleotide Mutations in Rice OsMsh6 Mutants

    Directory of Open Access Journals (Sweden)

    Hairui Cui

    2017-01-01

    Full Text Available Genomic stability depends in part on an efficient DNA lesion recognition and correction by the DNA mismatch repair (MMR system. We investigated mutations arising spontaneously in rice OsMsh6 mutants by specific-locus amplified fragment sequencing. Totally 994 single-nucleotide mutations were identified in three mutants and on average the mutation density is about 1/136.72 Kb per mutant line. These mutations were relatively randomly distributed in genome and might be accumulated in generation-dependent manner. All possible base transitions and base transversions could be seen and the ratio of transitions to transversions was about 3.12. We also observed the nearest-neighbor bias around the mutated base. Our data suggests that OsMsh6 (LOC_Os09g24220 is important in ensuring genome stability by recognizing mismatches that arise spontaneously and provides useful information for investigating the function of the OsMsh6 gene in DNA repair and exploiting MMR mutants in rice induced mutation breeding.

  7. Genetic Analysis and Mapping of TWH Gene in Rice Twisted Hull Mutant

    Institute of Scientific and Technical Information of China (English)

    LI Jin-bo; XIA Ming-yuan; WAN Bing-liang; DU Xue-shu; ZHA Zhong-ping; YU Da-zhao; QI Hua-xiong

    2009-01-01

    A mutant with twisted hulls was found in a breeding population of rice (Oryza sativa L.). The mutant shows less grain weight and inferior grain quality in addition to twisted hulls. Genetic analysis indicated that the phenotype of mutant was controlled by a single recessive gene (temporarily designated as TWH). To map the TWH gene, an F2 population was generated by crossing the twh mutant to R725, an indica rice variety with normal hulls. For bulked segregant analysis, the bulk of mutant plants was prepared by mixing equal amount of plant tissue from 10 twisted-hull plants and the bulk of normal plants was obtained by pooling equal amount tissue of 10 normal-hull plants. Two hundred and seven pairs of simple sequence repeat (SSR) primers, which are distributed on 12 rice chromosomes, were used for polymorphism analysis of the parents and the two bulks. The TWH locus was initially mapped close to the SSR marker RM526 on chromosome 2. Therefore, further mapping was performed using 50 pairs of SSR primers around the marker RM526. The TWH was delimited between the SSR markers RM14128 and RM208 on the long arm of chromosome 2 at the genetic distances of 1.4 cM and 2.7 cM, respectively. These results provide the foundation for further fine mapping, cloning and functional analysis of the TWH gene.

  8. Identification of altered metabolic pathways of γ-irradiated rice mutant via network-based transcriptome analysis.

    Science.gov (United States)

    Hwang, Sun-Goo; Kim, Dong Sub; Hwang, Jung Eun; Park, Hyeon Mi; Jang, Cheol Seong

    2015-12-01

    In order to develop rice mutants for crop improvement, we applied γ-irradiation mutagenesis and selected a rice seed color mutant (MT) in the M14 targeting-induced local lesions in genome lines. This mutant exhibited differences in germination rate, plant height, and root length in seedlings compared to the wild-type plants. We found 1645 different expressed probes of MT by microarray hybridization. To identify the modified metabolic pathways, we conducted integrated genomic analysis such as weighted correlation network analysis with a module detection method of differentially expressed genes (DEGs) in MT on the basis of large-scale microarray transcriptional profiling. These modules are largely divided into three subnetworks and mainly exhibit overrepresented gene ontology functions such as oxidation-related function, ion-binding, and kinase activity (phosphorylation), and the expressional coherences of module genes mainly exhibited in vegetative and maturation stages. Through a metabolic pathway analysis, we detected the significant DEGs involved in the major carbohydrate metabolism (starch degradation), protein degradation (aspartate protease), and signaling in sugars and nutrients. Furthermore, the accumulation of amino acids (asparagine and glutamic acid), sucrose, and starch in MT were affected by gamma rays. Our results provide an effective approach for identification of metabolic pathways associated with useful agronomic traits in mutation breeding.

  9. Mapping and characterization of a tiller-spreading mutant lazy-2 in rice

    Institute of Scientific and Technical Information of China (English)

    LI Peijin; ZENG Dali; LIU Xinfang; XU Dan; GU Dai; LI Jiayang; QIAN Qian

    2003-01-01

    Tiller angle of rice is an important agronomic trait that contributes to breed new varieties with ideal architecture. In this study, we report mapping and characterization of a rice mutant defective in tiller angle. At the seedling stage, the newly developed tillers of the mutant plants grow with a large angle that leads to a "lazy" phenotype at the mature stage. Genetic analysis indicates that this tiller-spreading phenotype is controlled by one recessive gene that is allelic to a reported mutant la. Therefore, the mutant was named la-2 and la renamed la-1. To map and clone LA, we constructed a large mapping population. Genetic linkage analysis showed that the LA gene is located between 2 SSR markers RM202 and RM229. By using the 6 newly-developed molecular markers, the LA gene was placed within a 0.4 cM interval on chromosome 11, allowing us to clone LA and study the mechanism that controls rice tiller angle at the molecular level.

  10. Analysis of genes specifically expressed under salt stress in salt-tolerant mutant of rice by using DDRT-PCR technique

    Institute of Scientific and Technical Information of China (English)

    张弛; 陈受宜

    1996-01-01

    Differential display reverse transcription-PCR (DDRT-PCR) technique was used to identify those genes that are expressed differentially between wild type rice variety 77-170 (Oryza Sativa vas Japonica) and its salt-tolerant mutant (M-20) under salt stress. Totally 13 salt-inducible cDNA fragments of 200-600 bp were identified and doned, and were designated as SIGR1 - SIGR13 (salt-induced gene in rice). Northern blot analysis showed that expression of SIGR6 and SIGR8 was salt-inducible in both wild type and mutant, and expression of SIGR12 in M-20 was much higher than that in 77-170 under salt stress. It was also shown that expression of SIGR3, SIGR4, SIGR7, SIGRIO and SIGR13 was salt-inducible, and the genes were highly homologous with Rab1d which was an ABA-inducible gene of rice. The great potential application of DDRT-PCR technique in plant molecular biology research may promote the investigation of expression of salt-induced protein in rice.

  11. Blast-Resistance Inheritance of Space-Induced Rice Lines and Their Genomic Polymorphism by Microsatellite Markers

    Institute of Scientific and Technical Information of China (English)

    XIAO Wu-ming; YANG Qi-yun; CHEN Zhi-qiang; WANG Hui; GUO Tao; LIU Yong-zhu; ZHU Xiao-yuan

    2009-01-01

    To understand the resistance inheritance basis of space-induced rice lines to blast,and to probe mutants'genomic DNA polymorphism compared with ground control by microsatellite markers,three space-induced lines were crossed with a highly susceptible variety LTH,and their F1 and F2 populations were inoculated by two representative blast isolates with broad pathogenicity to analyze their resistance inheritance basis.Meanwhile three mutant lines and the ground control were analyzed by 225 rice SSR(simple sequence repeat)primer pairs selected throughout the 12 chromosomes of whole rice genome,to scan the mutagenesis in genome of the mutant lines.The results indicated the blast-resistant genes harbored in these mutant lines were dominant.It was demonstrated that the resistance of mutant H1 to isolate GD0193 and GD3286 was controlled by a single gene,respectively;while mutants H2 and H3 were controlled by two pairs of major genes against isolate GD3286 and H2 showed complicated genetic mechanism to isolate GD0193.H3's resistance to isolate GD0193 was verified to be controlled by a single gene.According to the results of SSR analysis,three mutant lines showed different mutant rates as compared with the ground control,and the mutant rates also varied.Resistance genes can be induced from rice by space mutation,and different genomic variations were detected in blast-resistant lines.

  12. Identification of Heading Date Six (Hd6 Gene Derived from Rice Mutant Varieties

    Directory of Open Access Journals (Sweden)

    Aryanti Aryanti

    2017-04-01

    Full Text Available Genes which were associated with flowering time to indicate the early maturity is known as heading date (Hd. Heading date six (Hd6 gene was identified from rice mutant varieties were Atomita 2, Atomita 3, Atomita 4, Bestari, Cilosari, Diah Suci, Sidenuk, Kahayan, Mayang, Meraoke, Mira-1, Pandan Putri, Superwin, Suluttan Unsrat 1, Suluttan Unsrat 2, Winongo, Woyla, Yuwono, while the rice var. Nipponbare was used as a positive control. All of rice mutant varieties derived from mutation induction by the dose of 0.2 kGy. The aim of this experiment was to find out the data base of mutant varieties which could be used as parent material with earlier maturity trait genetically. To obtain the DNA of plants, young leaves of each variety were extracted by liquid nitrogen, and then lysis and extracted by Kit Plant Genomic DNA. The amplification of DNA with 7 primers of Hd6 conducted of 40 cycles by PCR and were continues to separated by 1 % agarose. The results were shown that the rice Mira-1 and Bestari varieties obtained from mutation of Cisantana highly different from one to another on 7 primers of Hd6 used. Mayang variety from mutation of cross breeding between Cilosari and IR64, Pandan putri from Pandan wangi and Woyla from mutation of cross breeding from Atomita 2 and IR64 were highly different with those of their parents. Identification of Hd6 gene on Sidenuk variety was shown the same bands pattern with Nipponbare as control positive toward all primers used, this variety would be better for earlier maturity parent material compared to others. The information could be useful for breeding programs aiming to develop early maturing widely adaptive and high yielding rice cultivars.

  13. Genetic Analysis and Gene Mapping of Short Root Mutant Rice ksr1

    Institute of Scientific and Technical Information of China (English)

    NING Yong-qiang; DING Wo-na; ZHU Shi-hua; Yu Hang-wei; Yu Hang; Lu Kai-xing

    2011-01-01

    A short root mutant ksr1 with the Kasalath background was isolated from an EMS-mutagenized population in rice.The root length of 6-day-old ksr1 seedlings was only about 20% of the wild type.Genetic analysis indicated that the short root phenotype of ksr1 was controlled by a recessive mutation in a single nuclear-encoded gene.To map the ksr1 mutation,an F2 population was generated by crossing the ksr1 mutant with Nipponbare.The KSR1 locus was linked to the SSR marker RM1223 on rice chromosome 4.Eight new SSR markers and two InDet markers were developed around this marker.KSR1 gene was further mapped to a 155 kb region,flanked by the InDel marker 4-24725K and the SSR marker RM17182.

  14. Phenotypic analysis and molecular characterization of an allelic mutant of the D61 gene in rice

    Institute of Scientific and Technical Information of China (English)

    Yanan; Gao; Guangquan; Wang; Shoujiang; Yuan; Yanling; Qin; Jinfeng; Zhao; Yanpei; Zhang; Wenhui; Zhang; Xueyong; Li

    2014-01-01

    Brassinosteroids(BRs) are a class of plant-specific steroidal hormones that play important roles in multiple biological processes. In this paper, a classic rice mutant gsor300084,showing erect leaves and semi-dwarf stature, was characterized. Morphological analysis in darkness showed that the mesocotyl of the gsor300084 mutant did not elongate when grown in darkness. Coleoptile elongation and root growth were less affected by the exogenous application of brassinolide(BL), the most active form of BR, in gsor300084 than in the wild-type rice variety Matsumae. Lamina joint bending analysis also showed that gsor300084 was less sensitive to exogenous BL than Matsumae. These results suggested that the gsor300084 mutant is defective in BR sensitivity. Map-based cloning indicated that gsor300084 is a novel allelic mutant of the DWARF61(D61) gene, which encodes the putative BR receptor OsBRI1. A single-base mutation appears in the LRR domain of OsBRI1, changing the 444 th amino acid from tryptophan(W) to arginine(R). Subcellular localization analysis suggested that both the wild-type and mutant OsBRI1 protein are localized at the cytoplasmic membrane. Structure modeling revealed that the W444 R substitution may affect the perception of BRs by the LRR domain.

  15. Phenotypic analysis and molecular characterization of an allelic mutant of the D61 gene in rice

    Directory of Open Access Journals (Sweden)

    Yanan Gao

    2014-08-01

    Full Text Available Brassinosteroids (BRs are a class of plant-specific steroidal hormones that play important roles in multiple biological processes. In this paper, a classic rice mutant gsor300084, showing erect leaves and semi-dwarf stature, was characterized. Morphological analysis in darkness showed that the mesocotyl of the gsor300084 mutant did not elongate when grown in darkness. Coleoptile elongation and root growth were less affected by the exogenous application of brassinolide (BL, the most active form of BR, in gsor300084 than in the wild-type rice variety Matsumae. Lamina joint bending analysis also showed that gsor300084 was less sensitive to exogenous BL than Matsumae. These results suggested that the gsor300084 mutant is defective in BR sensitivity. Map-based cloning indicated that gsor300084 is a novel allelic mutant of the DWARF61 (D61 gene, which encodes the putative BR receptor OsBRI1. A single-base mutation appears in the LRR domain of OsBRI1, changing the 444th amino acid from tryptophan (W to arginine (R. Subcellular localization analysis suggested that both the wild-type and mutant OsBRI1 protein are localized at the cytoplasmic membrane. Structure modeling revealed that the W444R substitution may affect the perception of BRs by the LRR domain.

  16. Thermostability of photosynthesis in two new chlorophyll b-less rice mutants

    Institute of Scientific and Technical Information of China (English)

    LIN; Zhifang; PENG; Changlian; Xu; Xinlan; LIN; Guizhu; ZH

    2005-01-01

    Leaves of the two new chlorophyll b-less rice mutants VG28-1, VG30-5 and the wild type rice cv. Zhonghua 11 were subjected to temperatures 28, 36, 40, 44 and 48℃ in the dark for 30 min or gradually elevated temperature from 30℃ to 80℃ at 0.5℃/min. The thermostability of photosynthetic apparatus was estimated by the changes in chlorophyll fluorescence parameters, photosynthetic rate and pigment content, chloroplast ultrastructure and tissue location of H2O2 accumulation. There were different patterns of Fo-temperature curves between the Chl b-less mutants and the wild type plant, and the temperature of Fo rising threshold was shifted 3℃ lower in the Chl b-less mutants (48℃) than in the wild type (51℃). At temperature up to about 45℃, chloroplasts were swollen and thylakoid grana became misty accompanied with the complete loss of photosynthetic oxygen evolution in the two Chl b-less mutants, but chloroplast ultrastructure in the wild type showed no obvious alteration. After 55℃ exposure, the disordered thylakoid and significant H2O2 accumulation in leaves were found in the two Chl b-less mutants, whereas in the wild type plant, less H2O2 was accumulated and the swollen thylakoid still maintained a certain extent of stacking. A large extent of the changes in qP, NPQ and Fv/Fm was consistent with the Pn decreasing rate in the Chl b-less mutants during high temperature treatment as compared with the wild type. The results indicated that the Chl b-less mutants showed a tendency for higher thermosensitivity, and loss of Chl b in LHC II could lead to less thermostability of PSII structure and function. Heat damage to photosynthetic apparatus might be partially attributed to the internal oxidative stress produced at severely high temperature.

  17. Genetic Analysis and Molecular Mapping of Light-Sensitive Red-Root Mutant in Rice

    Institute of Scientific and Technical Information of China (English)

    ZHANG Jun-zhi; LIU Xiao; LI Chao; XIAO Ke; DONG Yan-jun

    2009-01-01

    The light-sensitive red-root mutant, designated as HG1, was newly observed from an indica rice variety, Nankinkodo, when seedlings were grown with roots exposed to natural light. The root color of the mutant began to turn slight-red when the roots were exposed to the light at the intensity of 29 μmol/(m2·s), then turned dark-red at the light intensity of 180 μmol/(m2·s), suggesting that the root color of the mutant was evidently sensitive to light. Furthermore, genetic analysis showed that the character of light-sensitive red-root of the HG1 mutant was controlled by a single dominant gene, tentatively designated as Lsr. With simple sequence repeat markers, Lsr gene was located between the markers RM252 and RM303 on chromosome 4 with the genetic distances of 9.8 cM and 6.4 cM, respectively. These results could be useful for fine mapping and cloning of Lsr gene in rice.

  18. Identification of variability for agronomically important traits in rice mutant families

    Directory of Open Access Journals (Sweden)

    Viviane Kopp da Luz

    2016-03-01

    Full Text Available ABSTRACT The increase of yield potential in new rice (Oryza sativa L. varieties has been a major challenge for genetic improvement. The generation of mutants, followed by their characterization, constitutes a great possibility to isolate and select genes and genotypes that present agronomic traits of interest. This study aimed to evaluate the effect of chemical mutagen ethyl methane sulphonate (EMS on agronomic traits in 340 M3 families of rice derived from BRS Querência cultivar. Seeds from each family and the original genotype BRS Querência were sown in the experimental field, and the characters main panicle length, main panicle weight, main panicle grain weight, flag leaf width and plant height were evaluated. Data were subjected to analysis of variance (p ≤ 0.05, and a comparison of means was carried out by Dunnett's test at 5% significance. The results show that there is genetic variability among the mutant families, suggesting that the mutagen EMS at 1.5% is effective for generating mutants for all assessed traits. Among the characters, plant height was the most affected by the mutagen, which provided an increase in the character. For the main panicle length character, seven families showed means above the control; for main panicle weight and grain weight, four and six mutant families were observed, respectively, with a superior performance in comparison to the control.

  19. Characterization and mapping of a white panicle mutant gene in rice

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    A spontaneous white panicle mutant was found from the F6 progenies of an indica/japonica cross. The mutant exhibits white stripes on its basal leaves while the panicles, rachis and pedicel are milky white colored at flowering stage. Genetic analysis in an F2 population from the cross of Zhi7/white panicle mutant indicates that the white panicle phenotype is controlled by a single recessive nuclear gene, tentatively termed as wp(t). Using microsatellite markers, the wp(t) gene was anchored between the markers of SSR101 and SSR63.9 with a map distance of 2.3 and 0.8 cM, respectively, and co-segregated with the marker of SSR17 on rice chromosome 1.

  20. A rice mutant displaying a heterochronically elongated internode carries a 100 kb deletion

    Institute of Scientific and Technical Information of China (English)

    Mika Hayashi-Tsugane; Masahiko Maekawa; Qian Qian; Hirokazu Kobayashi; Shigeru Iida; Kazuo Tsugane

    2011-01-01

    We have isolated a recessive rice mutant,designated as indeterminate growth(ing),which displays creeping and apparent heterochronic phenotypes in the vegetative period with lanky and winding culms.Rough mapping and subsequent molecular characterization revealed that the ing mutant carries a large deletion,which corresponds to a 103 kb region in the Nipponbare genome,containing nine annotated genes on chromosome 3.Of these annotated genes,the SLRI gene encoding a DELLA protein is the only one that is well characterized in its function,and its null mutation,which is caused by a single base deletion in the middle of the intronless SLR1 gene,confers a slender phenotype that bears close resemblance to the ing mutant phenotype.The primary cause of the ing mutant phenotype is the deletion of the SLR1 gene,and the ing mutant appears to be the first characterized mutant having the entire SLRI sequence deleted.Our results also suggest that the deleted region of 103 kb does not contain an indispensable gene,whose dysfunction must result in a lethal phenotype.

  1. Rice Brittleness Mutants: A Way to Open the 'Black Box' of Monocot Cell Wall Biosynthesis

    Institute of Scientific and Technical Information of China (English)

    Baocai Zhang; Yihua Zhou

    2011-01-01

    Rice is a model organism for studying the mechanism of cell wall biosynthesis and remolding in Gramineae.Mechanical strength is an important agronomy trait of rice(Oryza sativa L.)plants that affects crop lodging and grain yield.As a prominent physical property of cell walls,mechanical strength reflects upon the structure of different wall polymers and how they interact.Studies on the mechanisms that regulate the mechanical strength therefore consequently results in uncovering the genes functioning in cell wall biosynthesis and remodeling.Our group focuses on the study of isolation of brittle culm(bc)mutants and characterization of their corresponding genes.To date,several bc mutants have been reported.The identified genes have covered several pathways of cell wall biosynthesis,revealing many secrets of monocot cell wall biosynthesis.Here,we review the progress achieved in this research field and also highlight the perspectives in expectancy.All of those lend new insights into mechanisms of cell wall formation and are helpful for harnessing the waste rice straws for biofuel production.

  2. Barnyard grass-induced rice allelopathy and momilactone B.

    Science.gov (United States)

    Kato-Noguchi, Hisashi

    2011-07-01

    Here, we investigated chemical-mediated interaction between crop and weeds. Allelopathic activity of rice seedlings exhibited 5.3-6.3-fold increases when rice and barnyard grass seedlings were grown together, where there may be the competitive interference between rice and barnyard grass for nutrients. Barnyard grass is one of the most noxious weeds in rice cultivation. The momilactone B concentration in rice seedlings incubated with barnyard grass seedlings was 6.9-fold greater than that in rice seedlings incubated independently. Low nutrient growth conditions also increased allelopathic activity and momilactone B concentrations in rice seedlings. However, the increases in the low nutrient-induced allelopathic activity and momilactone B concentration were much lower than those in barnyard grass-induced allelopathic activity and momilactone B concentration. Root exudates of barnyard grass seedlings increased allelopathic activity and momilactone B concentration in rice seedlings at concentrations greater than 30 mg/L of the root exudates, and increasing the exudate concentration increased the activity and momilactone B concentration. Therefore, barnyard grass-induced allelopathic activity of rice seedlings may be caused not only by nutrient competition between two species, but also by components in barnyard grass root exudates. As momilactone B shows strong allelopathic activities, barnyard grass-induced allelopathic activity of rice may be due to the increased concentration of momilactone B in rice seedlings. The present research suggests that rice may respond to the presence of neighboring barnyard grass by sensing the components in barnyard grass root exudates and increasing allelopathic activity by production of elevated concentration of momilactone B. Thus, rice allelopathy may be one of the inducible defense mechanisms by chemical-mediated plant interaction between rice and barnyard grass, and the induced-allelopathy may provide a competitive advantage for

  3. Identification and Genetic Mapping of a Lesion Mimic Mutant in Rice

    Institute of Scientific and Technical Information of China (English)

    MA Jian-yang; CHEN Sun-lu; ZHANG Jian-hui; DONG Yan-jun; TENG Sheng

    2012-01-01

    A lesion mimic stripe mutant,designated as Ims1 (lesion mimic stripe 1),was obtained from the M2 progeny of a 60Co Y-radiation treated japonica rice variety Jiahua 1.The Ims1 mutant displayed propagation type lesions across the whole growth and developmental stages.Physiology and histochemistry analysis showed that the mutant exhibited a phenotype of white stripe when grown under high temperature (30 ℃),and the lesion mimic caused by programmed cell death under low temperature (20 ℃).The genetic analysis indicated that this lesion-mimic phenotype is controlled by a single locus recessive nuclear gene.Furthermore,by using simple sequence repeat markers and an F2 segregating population derived from two crosses of Ims1 × 93-11 and Ims1 × Pei'ai 64S,the Ims1 gene was mapped between markers Indel1 and MM0112-4 with a physical distance of 400 kb on chromosome 6 in rice.

  4. Organ fusion and defective shoot development in oni3 mutants of rice

    Science.gov (United States)

    Akiba, Takafumi; Hibara, Ken-Ichiro; Kimura, Fumiko; Tsuda, Katsutoshi; Shibata, Kiko; Ishibashi, Mayu; Moriya, Chihiro; Nakagawa, Kiyotaka; Kurata, Nori; Itoh, Jun-Ichi; Ito, Yukihiro

    2014-01-01

    Maintenance of organ separation is one of the essential phenomena for normal plant development. We have identified and analyzed ONION3 (ONI3), which is required for avoiding organ fusions in rice. Loss-of-function mutations of ONI3, which were identified as mutants with ectopic expression of KNOX genes in leaves and morphologically resembling KNOX overexpressors, showed abnormal organ fusions in developing shoots. The mutant seedlings showed fusions between neighboring organs and also within an organ; they stopped growing soon after germination and subsequently died. ONI3 was shown to encode an enzyme that is most similar to Arabidopsis HOTHEAD and is involved in biosynthesis of long-chain fatty acids. Expression analyses showed that ONI3 was specifically expressed in the outermost cell layer in the shoot apex throughout life cycle, and the oni3 mutants had an aberrant outermost cell layer. Our results together with previous studies suggest that long-chain fatty acids are required for avoiding organ fusions and promoting normal shoot development in rice. PMID:24192297

  5. Morphology and mapping analysis of rice (Oryza sativa L.) clustered spikelets (Cl) mutant

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    The rice clustered spikelets (Cl) mutant exhibits a phenotype that most of branch apical have 2-3 spikelets clustered together. SEM (scanning electron microscope) observation suggested that the Cl gene controlled branch apical development, and influenced the terminal spikelets elongation. The spikelet number was reduced in mutant, indicating that Cl may also have an effect on spikelet number. To map Cl locus, two F2 mapping populations derived from the crosses between the Cl and ZhongHua11, and Cl and ZheFu802 were constructed, respectively. The Cl locus was roughly mapped between two CAPS markers, CK0214 and SS0324. A further fine mapping analysis showed that the Cl locus was mapped between makers R0674E and Cl2560, with genetic distances of 0.2 and 2.1 cM, respectively. Then we found a PAC contig spanning Cl locus, the region was delimited to 196 kb. This result was useful for cloning of the Cl gene. Allelism test demonstrated that Cl was allelic to Cl2, another rice clustered spikelets mutant.

  6. Genetic analysis and preliminary gene mapping of a novel seeding yellowleaf mutant in rice

    Directory of Open Access Journals (Sweden)

    GE Shaoxing

    2013-02-01

    Full Text Available A novel seeding yellow-leaf mutant, syl11(seedling yellow leaf 11,was found in rice mutant pool from japonica variety,Nipponbare,treated by 60Coγ-radiation.In comparison with the wild-type parent (Nipponbare,the mutant displayed the phenotype of yellow on the 2nd and 3rd leaf and began to turn green from its top before they completely expanded besides the significantly decreased contents of photosynthetic pigments,but all leaves become normal green after the 4-leaf-old stage.The genetic analysis indicated that the mutant trait was controlled by a single recessive nuclear gene.In addition,with the help of SSR,new developed InDel markers and F2 segregating population derived from the cross between the syl11 mutant and Peiai64S,the syl11 was mapped between RM26652(on the long armand ID11974(closing to the centromereon chromosome 11,with a genetic distance of 0.5cM and 0.7cM,respectively.

  7. Genetic analysis and preliminary gene mapping of a novel seeding yellow leaf mutant in rice

    Directory of Open Access Journals (Sweden)

    GE Shaoxing

    2013-02-01

    Full Text Available A novel seeding yellow-leaf mutant, syl11(seedling yellow leaf 11,was found in rice mutant pool from japonica variety,Nipponbare,treated by 60Coγ-radiation.In comparison with the wild-type parent (Nipponbare,the mutant displayed the phenotype of yellow on the 2nd and 3rd leaf and began to turn green from its top before they completely expanded besides the significantly decreased contents of photosynthetic pigments,but all leaves become normal green after the 4-leaf-old stage.The genetic analysis indicated that the mutant trait was controlled by a single recessive nuclear gene.In addition,with the help of SSR,new developed InDel markers and F2 segregating population derived from the cross between the syl11 mutant and Peiai64S,the syl11 was mapped between RM26652(on the long armand ID11974(closing to the centromereon chromosome 11,with a genetic distance of 0.5cM and 0.7cM,respectively.

  8. Genetic analysis and gene mapping of a narrow leaf mutant in rice (Oryza sativa L.)

    Institute of Scientific and Technical Information of China (English)

    WANG DeKai; LIU HeQin; LI KeLei; LI SuJuan; TAO YueZhi

    2009-01-01

    A narrow leaf mutant was obtained after T-DNA transformation conducted on a rice variety Zhonghua 11. Several abnormal morphological characteristics, including semi-dwarf, delayed flowering time, narrow and inward rolling leaves, and lower seed-setting, were observed. The rate of net photosynthesis (un-der saturate light) of flag leaves in the mutant was significantly lower than that of the wild type. More-over, the leaf transpiration rate and stomatal conductance in the mutant flag leaf were lower than those of the wild type at the grain filling stage. It was found that the mutant phenotype was not caused by the T-DNA insertion. Genetic analysis showed that the mutant was controlled by a single recessive gene,designated as nal3(t). A genetic linkage map was constructed using a large F2 mapping population de-rived from a cross between nal3(t) and an indica variety Longtefu B with 6 polymorphic markers on chromosome 12 identified from 366 SSR markers by the BAS method. Gene nal3(t) was mapped be-tween the markers RM7018 and RM3331. Fine mapping of nal3(t) locus was conducted with 22 newly developed STS markers based on the sequence diversity around the region harboring nal3(t) between Nipponbare and 93-11, and nal3(f) was finally mapped to a 136-kb region between the STS markers NS10 and RH12-8.

  9. MutMap+: genetic mapping and mutant identification without crossing in rice.

    Directory of Open Access Journals (Sweden)

    Rym Fekih

    Full Text Available Advances in genome sequencing technologies have enabled researchers and breeders to rapidly associate phenotypic variation to genome sequence differences. We recently took advantage of next-generation sequencing technology to develop MutMap, a method that allows rapid identification of causal nucleotide changes of rice mutants by whole genome resequencing of pooled DNA of mutant F2 progeny derived from crosses made between candidate mutants and the parental line. Here we describe MutMap+, a versatile extension of MutMap, that identifies causal mutations by comparing SNP frequencies of bulked DNA of mutant and wild-type progeny of M3 generation derived from selfing of an M2 heterozygous individual. Notably, MutMap+ does not necessitate artificial crossing between mutants and the wild-type parental line. This method is therefore suitable for identifying mutations that cause early development lethality, sterility, or generally hamper crossing. Furthermore, MutMap+ is potentially useful for gene isolation in crops that are recalcitrant to artificial crosses.

  10. MutMap+: Genetic Mapping and Mutant Identification without Crossing in Rice

    Science.gov (United States)

    Abe, Akira; Natsume, Satoshi; Yaegashi, Hiroki; Sharma, Shailendra; Sharma, Shiveta; Kanzaki, Hiroyuki; Matsumura, Hideo; Saitoh, Hiromasa; Mitsuoka, Chikako; Utsushi, Hiroe; Uemura, Aiko; Kanzaki, Eiko; Kosugi, Shunichi; Yoshida, Kentaro; Cano, Liliana; Kamoun, Sophien; Terauchi, Ryohei

    2013-01-01

    Advances in genome sequencing technologies have enabled researchers and breeders to rapidly associate phenotypic variation to genome sequence differences. We recently took advantage of next-generation sequencing technology to develop MutMap, a method that allows rapid identification of causal nucleotide changes of rice mutants by whole genome resequencing of pooled DNA of mutant F2 progeny derived from crosses made between candidate mutants and the parental line. Here we describe MutMap+, a versatile extension of MutMap, that identifies causal mutations by comparing SNP frequencies of bulked DNA of mutant and wild-type progeny of M3 generation derived from selfing of an M2 heterozygous individual. Notably, MutMap+ does not necessitate artificial crossing between mutants and the wild-type parental line. This method is therefore suitable for identifying mutations that cause early development lethality, sterility, or generally hamper crossing. Furthermore, MutMap+ is potentially useful for gene isolation in crops that are recalcitrant to artificial crosses. PMID:23874658

  11. Characterization and mapping of a novel light-dependent lesion mimic mutant Imm6 in rice (Oryza sativa L.)

    Institute of Scientific and Technical Information of China (English)

    XIAO Gui-qing[1,2; ZHANG Hal-wen[3; LU Xiang-yang[1,2; HUANG Rong-feng[3

    2015-01-01

    A novel rice lesion mimic mutant (LMM) was isolated from an ethane methyl sulfonate (EMS)-induced 02428 mutant bank. The mutant, tentatively designated as Imm6, develops necrotic lesions in the whole growth period along with changes in several important agronomic traits. We found that the initiation of the lesions was induced by light and cell death occurred in Imm6 accompanied with accumulation of reactive oxygen species (ROS). The lower chlorophyll content, soluble protein content and superoxide dismutase (SOD) activity, the higher malondialdehyde (MDA) content were detected in Imm6 than in the wild type (WT). Moreover, the observation by transmission electronic microscope (TEM) demonstrated that some organelles were damaged and the stroma lamella of chloroplast was irregular and loose in mesophyll cell of Imm6. In addition, Imm6 was more resistant than WT to rice blast fungus Magnaporthe grisea infection, which was consistent with increased expression of four genes involved in the defense-related reaction. Genetic analysis showed that mutant trait of Imm6 is inherited as a monogenic recessive nuclear gene located on the long arm of chromosome 6. Using simple sequence repeat (SSR) markers, the target gene was finally delimited to an interval of 80.8 kb between markers MM2359 and MM2370, containing 7 annotated genes. Taken together, our results provide the information to identify a new gene involved in rice lesion mimic, which will be helpful in clarifying the mechanism of cell death and disease resistance in rice.

  12. Isolation and characterisation of a dwarf rice mutant exhibiting defective gibberellins biosynthesis.

    Science.gov (United States)

    Ji, S H; Gururani, M A; Lee, J W; Ahn, B-O; Chun, S-C

    2014-03-01

    We have isolated a severe dwarf mutant derived from a Ds (Dissociation) insertion mutant rice (Oryza sativa var. japonica c.v. Dongjin). This severe dwarf phenotype, has short and dark green leaves, reduced shoot growth early in the seedling stage, and later severe dwarfism with failure to initiate flowering. When treated with bioactive GA3 , mutants are restored to the normal wild-type phenotype. Reverse transcription PCR analyses of 22 candidate genes related to the gibberellin (GA) biosynthesis pathway revealed that among 22 candidate genes tested, a dwarf mutant transcript was not expressed only in one OsKS2 gene. Genetic analysis revealed that the severe dwarf phenotype was controlled by recessive mutation of a single nuclear gene. The putative OsKS2 gene was a chromosome 4-located ent-kaurene synthase (KS), encoding the enzyme that catalyses an early step of the GA biosynthesis pathway. Sequence analysis revealed that osks2 carried a 1-bp deletion in the ORF region of OsKS2, which led to a loss-of-function mutation. The expression pattern of OsKS2 in wild-type cv Dongjin, showed that it is expressed in all organs, most prominently in the stem and floral organs. Morphological characteristics of the dwarf mutant showed dramatic modifications in internal structure and external morphology. We propose that dwarfism in this mutant is caused by a point mutation in OsKS2, which plays a significant role in growth and development of higher plants. Further investigation on OsKS2 and other OsKS-like proteins is underway and may yield better understanding of the putative role of OsKS in severe dwarf mutants.

  13. Antimicrobial Activity of UV-Induced Phenylamides from Rice Leaves

    Directory of Open Access Journals (Sweden)

    Hye Lin Park

    2014-11-01

    Full Text Available Rice produces a wide array of phytoalexins in response to pathogen attacks and UV-irradiation. Except for the flavonoid sakuranetin, most phytoalexins identified in rice are diterpenoid compounds. Analysis of phenolic-enriched fractions from UV-treated rice leaves showed that several phenolic compounds in addition to sakuranetin accumulated remarkably in rice leaves. We isolated two compounds from UV-treated rice leaves using silica gel column chromatography and preparative HPLC. The isolated phenolic compounds were identified as phenylamide compounds: N-trans-cinnamoyltryptamine and N-p-coumaroylserotonin. Expression analysis of biosynthetic genes demonstrated that genes for arylamine biosynthesis were upregulated by UV irradiation. This result suggested that phenylamide biosynthetic pathways are activated in rice leaves by UV treatment. To unravel the role of UV-induced phenylamides as phytoalexins, we examined their antimicrobial activity against rice fungal and bacterial pathogens. N-trans-Cinnamoyltryptamine inhibited the growth of rice brown spot fungus (Bipolaris oryzae. In addition to the known antifungal activity to the blast fungus, sakuranetin had antimicrobial activity toward B. oryzae and Rhizoctonia solani (rice sheath blight fungus. UV-induced phenylamides and sakuranetin also had antimicrobial activity against rice bacterial pathogens for grain rot (Burkholderia glumae, blight (Xanthomonas oryzae pv. oryzae and leaf streak (X. oryzae pv. oryzicola diseases. These findings suggested that the UV-induced phenylamides in rice are phytoalexins against a diverse array of pathogens.

  14. Identiifcation and Genetic Analysis of a Novel Rice Spotted-Leaf Mutant with Broad-Spectrum Resistance to Xanthomonas oryzae pv. oryzae

    Institute of Scientific and Technical Information of China (English)

    SHEN Hai-chao; SHI Yong-feng; FENG Bao-hua; WANG Hui-mei; XU Xia; HUANG Qi-na; LÜ Xiang-guang; WU Jian-li

    2014-01-01

    A spotted-leaf mutant of rice HM143 was isolated from an EMS-induced IR64 mutant bank. Brown lesions randomly distributed on leaf blades were observed about 3 wk after sowing. The symptom lasted for the whole plant growth duration. Histochemical analysis indicated that cell death occurred in and around the site of necrotic lesions accompanied with accumulation of hydrogen hyperoxide. Agronomic traits were largely similar to the wild type IR64 except seed setting rate and 1 000-grain weight which were signiifcantly decreased in the mutant. Disease resistance of the mutant to multiple races of Xanthomonas oryzae pv. oryzae was signiifcantly enhanced. Genetic analysis showed that the mutation was controlled by a single recessive gene, tentatively termed splHM143. In addition, using molecular markers and 1 023 mutant type individuals from an F2 segregating population derived from the cross HM143/R9308, the spotted-leaf gene was ifnally delimited to an interval of 149 kb between markers XX25 and ID40 on the long arm of chromosome 4. splHM143 is likely a novel rice spotted-leaf gene since no other similar genes have been identiifed near the chromosomal region.

  15. Characterization of SSIIIa-deficient mutants of rice: the function of SSIIIa and pleiotropic effects by SSIIIa deficiency in the rice endosperm.

    Science.gov (United States)

    Fujita, Naoko; Yoshida, Mayumi; Kondo, Tomonori; Saito, Kaori; Utsumi, Yoshinori; Tokunaga, Takashi; Nishi, Aiko; Satoh, Hikaru; Park, Jin-Hee; Jane, Jay-Lin; Miyao, Akio; Hirochika, Hirohiko; Nakamura, Yasunori

    2007-08-01

    Starch synthase IIIa (SSIIIa)-deficient rice (Oryza sativa) mutants were generated using retrotransposon insertion and chemical mutagenesis. The lowest migrating SS activity bands on glycogen-containing native polyacrylamide gel, which were identified to be those for SSIIIa, were completely absent in these mutants, indicating that they are SSIIIa null mutants. The amylopectin B(2) to B(4) chains with degree of polymerization (DP) >/= 30 and the M(r) of amylopectin in the mutant were reduced to about 60% and 70% of the wild-type values, respectively, suggesting that SSIIIa plays an important part in the elongation of amylopectin B(2) to B(4) chains. Chains with DP 6 to 9 and DP 16 to 19 decreased while chains with DP 10 to 15 and DP 20 to 25 increased in the mutants amylopectin. These changes in the SSIIIa mutants are almost opposite images of those of SSI-deficient rice mutant and were caused by 1.3- to 1.7-fold increase of the amount of SSI in the mutants endosperm. Furthermore, the amylose content and the extralong chains (DP >/= 500) of amylopectin were increased by 1.3- and 12-fold, respectively. These changes in the composition in the mutants starch were caused by 1.4- to 1.7-fold increase in amounts of granules-bound starch synthase (GBSSI). The starch granules of the mutants were smaller with round shape, and were less crystalline. Thus, deficiency in SSIIIa, the second major SS isozyme in developing rice endosperm affected the structure of amylopectin, amylase content, and physicochemical properties of starch granules in two ways: directly by the SSIIIa deficiency itself and indirectly by the enhancement of both SSI and GBSSI gene transcripts.

  16. Comparative metabolomic analysis of wild type and mads3 mutant rice anthers

    Institute of Scientific and Technical Information of China (English)

    Guorun Qu; Sheng Quan; Palash Mondol; Jie Xu; Dabing Zhang; Jianxin Shi

    2014-01-01

    Rice (Oryza sativa L.) MADS3 transcription factor regulates the homeostasis of reactive oxygen species (ROS) during late anther development, and one MADS3 mutant, mads3-4, has defective anther wal s, aborted microspores and complete male sterility. Here, we report the untargeted metabolomic analysis of both wild type and mads3-4 mature anthers. Mutation of MADS3 led to an unbalanced redox status and caused oxidative stress that damages lipid, protein, and DNA. To cope with oxidative stress in mads3-4 anthers, soluble sugars were mobilized and carbohydrate metabolism was shifted to amino acid and nucleic acid metabolism to provide substrates for the biosynthesis of antioxidant proteins and the repair of DNA. Mutation of MADS3 also affected other aspects of rice anther development such as secondary metabolites associated with cuticle, cellwal , and auxin metabolism. Many of the discovered metabolic changes in mads3-4 anthers were corroborated with changes of expression levels of correspond-ing metabolic pathway genes. Altogether, this comparative metabolomic analysis indicated that MADS3 gene affects rice anther development far beyond the ROS homeostasis regulation.

  17. A proteomic study of rice cultivar TNG67 and its high aroma mutant SA0420.

    Science.gov (United States)

    Lin, Da-Gin; Chou, Szu-Yi; Wang, Arthur Z; Wang, Yi-Wen; Kuo, Shu-Ming; Lai, Chien-Chen; Chen, Liang-Jwu; Wang, Chang-Sheng

    2014-01-01

    Fragrance is a very important economic trait for rice cultivars. To identify the aroma genes in rice, we performed a proteomics analysis of aroma-related proteins between Tainung 67 (TNG67) and its high aroma mutant SA0420. Seventeen of the differentially identified proteins were close related with the aroma phenotype of SA0420. Among them, 9 were found in leaves and 8 were found in grains. One protein (L3) was identified as the chloroplastic glyceraldehyde-3-phosphate dehydrogenase B (OsGAPDHB) which was less abundant in SA0420 than TNG67. Sequence analysis demonstrated that this protein in SA0420 carries a P425S mutation in the C-terminal extension domain, which might hinder the formation of holoenzyme, thereby changing the profile of aroma compounds. The protein profile of OsGAPDHB showed only a weak correlation to its transcription profile. This result indicated that the reduction of OsGAPDHB in SA0420 is regulated by post-translational processes and can only be analyzed by proteomics approach. Transgenic lines suppressing OsGAPDHB through RNAi harbored more fragrance than TNG67 but less than SA0420. With betaine-aldehyde dehydrogenase as the only fragrance gene identified in rice to date, OsGAPDHB may serve as the second protein known to contribute to the aroma phenotype.

  18. Water-deficit tolerant classification in mutant lines of indica rice

    Directory of Open Access Journals (Sweden)

    Suriyan Cha-um

    2012-04-01

    Full Text Available Water shortage is a major abiotic stress for crop production worldwide, limiting the productivity of crop species, especially in dry-land agricultural areas. This investigation aimed to classify the water-deficit tolerance in mutant rice (Oryza sativa L. spp. indica genotypes during the reproductive stage. Proline content in the flag leaf of mutant lines increased when plants were subjected to water deficit. Relative water content (RWC in the flag leaf of different mutant lines dropped in relation to water deficit stress. A decrease RWC was positively related to chlorophyll a degradation. Chlorophyll a , chlorophyll b , total chlorophyll , total carotenoids , maximum quantum yield of PSII , stomatal conductance , transpiration rate and water use efficiency in mutant lines grown under water deficit conditions declined in comparison to the well-watered, leading to a reduction in net-photosynthetic rate. In addition, when exposed to water deficit, panicle traits, including panicle length and fertile grains were dropped. The biochemical and physiological data were subjected to classify the water deficit tolerance. NSG19 (positive control and DD14 were identified as water deficit tolerant, and AA11, AA12, AA16, BB13, BB16, CC12, CC15, EE12, FF15, FF17, G11 and IR20 (negative control as water deficit sensitive, using Ward's method.

  19. Improvement of traditional local rice varieties through induced mutations using nuclear techniques

    Energy Technology Data Exchange (ETDEWEB)

    Pham Van Ro; Do Huu At [Cuu Long Delta Rice Research Institute (Viet Nam)

    2001-03-01

    'Improvement of local rice varieties for high yield, resistance to disease and insect pests (brown plant hopper and rice blast) and export quality through induced mutations for the Mekong Delta' started in 1993. After six years, it showed effecting on the field in the MD as well as at the south of Vietnam. TNDB-100 manifest very wide adaptation and yield stable variety. THDB is suitable for deepwater rice region, coastal area, where rice cultivation effected by acid sulphate and salinity conditions. Both varieties are good example for the method. Thank to good Co-operation from extension center from provinces, hundred classes of extension were organized to recommend to the farmers. And thank to the strongly supporting from IAEA so that nearly 400,000 ha of TNDB-100 occupied at the south of Vietnam as well as nearly 15,000 ha of THDB grown in the coastal as well as rainfed lowland rice areas at the South of Vietnam. To continue the rice improvement by this technique, seeds of six traditional local varieties were exposed under different dose of gamma rays to create new mutants. At present day hundred improved breeding lines were selected, a dozen of uniform lines were isolated and entranced the yield trail as well as regional testing program. From these improved varieties would be selected to contribute to the rice cultivation at the south of Vietnam in the next years. (author)

  20. Characterization and Fine Mapping of a Novel Rice Narrow Leaf Mutant nal9

    Institute of Scientific and Technical Information of China (English)

    Wei Li; Wenzhen Liu; Chao Wu; Guocheng Hu; Li Xing; Wenjing Qian; Huamin Si; Zongxiu Sun; Xingchun Wang; Yaping Fu

    2013-01-01

    A narrow leaf mutant was isolated from transgenic rice (Oryza sativa L.) lines carrying a T-DNA insertion. The mutant is characterized by narrow leaves during its whole growth period, and was named nal9 (narrow leaf 9). The mutant also has other phenotypes, such as light green leaves at the seedling stage, reduced plant height, a small panicle and increased tillering. Genetic analysis revealed that the mutation is controlled by a single recessive gene. A hygromycin resistance assay showed that the mutation was not caused by T-DNA insertion, so a map-based cloning strategy was employed to isolate the nal9 gene. The mutant individuals from the F2 generations of a cross between the nal9 mutant and Longtepu were used for mapping. With 24 F2 mutants, the nal9 gene was preliminarily mapped near the marker RM156 on the chromosome 3. New INDEL markers were then designed based on the sequence differences between japonica and indica at the region near RM156. The nal9 gene was finally located in a 69.3 kb region between the markers V239B and V239G within BAC OJ1212_C05 by chromosome walking. Sequence and expression analysis showed that an ATP-dependent Clp protease proteolytic subunit gene (ClpP) was most likely to be the nal9 gene. Furthermore, the nal9 mutation was rescued by transformation of the ClpP cDNA driven by the 35S promoter. Accordingly, the ClpP gene was identified as the NAL9 gene. Our results provide a basis for functional studies of NAL9 in future work.

  1. Research on the ultrafast fluorescence property of thylakoid membranes of the wild-type and mutant rice

    Institute of Scientific and Technical Information of China (English)

    任兆玉; 许晓明; 王水才; 辛越勇; 贺俊芳; 侯洵

    2003-01-01

    A high yielding rice variety mutant (Oryza sativa L., Zhenhui 249) with low chlorophyll b (Chl b) has been discovered in natural fields. It has a quality character controlled by a pair of recessive genes (nuclear gene). The partial loss of Chl b in content affects the efficiency of light harvest in a light harvest complex (LHC), thus producing the difference of the exciting energy transfer and the efficiency of photochemistry conversion between the mutant and wild-type rice in photosynthetic unit. The efficiency of utilizing light energy is higher in the mutant than that in the wildtype rice relatively. For further discussion of the above-mentioned difference and learning about the mechanism of the increase in the photochemical efficiency of the mutant, the pico-second resolution fluorescence spectrum measurement with delay-frame-scanning single photon counting technique is adopted. Thylakoid membranes of the mutant and the wild-type rice are excited by an Ar+ laser with a pulse width of 120ps, repetition rate of 4MHz and wavelength of 514nm. Compared with the time and spectrum property of exciting fluorescence, conclusions of those ultrafast dynamic experiments are: 1) The speeds of the exciting energy transferred in photo-system I are faster than that in photo-system II in both samples. 2) The speeds of the exciting energy transfer of mutant sample are faster than those of the wildtype. This might be one of the major reasons why the efficiency of photosynthesis is higher in mutant than that in the wild-type rice.

  2. Mapping of a rice thermosensitive genic male sterility gene from a TGMS mutant line

    Energy Technology Data Exchange (ETDEWEB)

    Vu Duc Quang; Nguyen Van Dong; Pham Ngoc Luong; Tran Duy Quy [Argicultural Genetics Institute, Hanoi (Viet Nam); Nguyen, Henry T. [Texas Tech Univ., Department of Plant and Soil Science, Lubbock TX (United States)

    2001-03-01

    At the Agricultural Genetics Institute (AGI), Hanoi, Vietnam, a number of thermo-sensitive genic male sterility (TGMS) homozygous rice lines have been developed by means of experimental mutagenesis followed by anther culture techniques. One of them (TGMS-1 indica mutant line) was used in this research. The critical temperature (at the period from pollen mother cell formation to the beginning of meiotic division) for TGMS-1 sterility was 24-25degC, below which the plants were fertile and above which the plants became sterile. Segregation analysis showed that the TGMS trait of the TGMS-1 mutant line was controlled by a single recessive gene. An F{sub 2} mapping population from a cross between TGMS-1 mutant line and CH1 (a fertile indica line) was developed for tagging and mapping the TGMS gene. From survey of 200 AFLP primer combinations in a bulked segregant analysis, 4 AFLP markers (E2/M5-200, E3/M16-400, E5/M12-600 and E5/M12-200) linked to TGMS-1 gene were identified and cloned. All except E2/M5-200 were found to be low-copy number sequences. The marker E5/M12-600 showed polymorphism in RFLP analysis and was closely linked to the TGMS gene at a distance of 3.3cM. This marker was subsequently mapped on chromosome 2 using doubled-haploid mapping populations derived from the crosses IR64xAzucena and CT9993xIR62666. Linkage of microsatellite marker RM27 with the TGMS gene further confirmed its location on chromosome 2. The closest marker, E5/M12-600, was sequenced so that a PCR marker can be developed for the use in marker-assisted breeding. The application of TGMS genes to the commercial two-line hybrid rice breeding system was discussed. (author)

  3. Transferring Translucent Endosperm Mutant Gene Wx-mq and Rice Stripe Disease Resistance Gene Stv-bi by Marker-Assisted Selection in Rice (Oryza sativa)

    Institute of Scientific and Technical Information of China (English)

    YAO Shu; CHEN Tao; ZHANG Ya-dong; ZHU Zhen; ZHAO Ling; ZHAO Qing-yong; ZHOU Li-hui; WANG Cai-lin

    2011-01-01

    A high-yielding japonica rice variety,Wuyunjing 7,bred in Jiangsu Province,China as a female parent was crossed with a Japanese rice variety Kantou 194,which carries a rice stripe disease resistance gene Stv-b1 and a translucent endosperm mutant gene Wx-mq.From F2 generations,a sequence characterized amplified region (SCAR) marker tightly linked with Stv-b1 and a cleaved amplified polymorphic sequence (CAPS) marker for Wx-mq were used for marker-assisted selection.Finally,a new japonica rice line,Ning 9108,with excellent agronomic traits was obtained by multi-generational selection on stripe disease resistance and endosperm appearance.The utilization of the markers from genes related to rice quality and disease resistance was helpful not only for establishing a marker-assisted selection system of high-quality and disease resistance for rice but also for providing important intermediate materials and rapid selection method for good quality,disease resistance and high yield in rice breeding.

  4. Growth deficiency of a Xanthomonas oryzae pv. oryzae fur mutant in rice leaves is rescued by ascorbic acid supplementation.

    Science.gov (United States)

    Subramoni, Sujatha; Sonti, Ramesh V

    2005-07-01

    Xanthomonas oryzae pv. oryzae causes bacterial leaf blight, a serious disease of rice. A mutation was isolated in the ferric uptake regulator (fur) gene of X. oryzae pv. oryzae and it was shown to result in the production of siderophores in a constitutive manner. The fur mutant is hypersensitive to the metallo-antibiotic streptonigrin, a phenotype that is indicative of intracellular free-iron overload, and also exhibits a slow growth phenotype on rich medium. The fur mutant is virulence deficient, hypersensitive to hydrogen peroxide, and exhibits reduced catalase activity. Exogenous supplementation with ascorbic acid (an antioxidant) rescues the growth deficiency of the fur mutant in rice leaves. The virulence deficiency of the X. oryzae pv. oryzae fur mutant is proposed to be due, at least in part, to an impaired ability to cope with the oxidative stress conditions that are encountered during infection.

  5. Fine Mapping and Cloning of Leafy Head Mutant Gene pla1-5 in Rice

    Directory of Open Access Journals (Sweden)

    Gong-neng FENG

    2013-09-01

    Full Text Available We identified a leafy head mutant pla1-5 (plastochron 1-5 from the progeny of japonica rice cultivar Taipei 309 treated with 60Co-γ ray irradiation. The pla1-5 mutant has a dwarf phenotype and small leaves. Compared with its wild type, pla1-5 has more leaves and fewer tillers, and it fails to produce normal panicles at the maturity stage. Genetic analysis showed that the pla1-5 phenotype is controlled by a single recessive nuclear gene. Using the map-based cloning strategy, we narrowed down the location of the target gene to a 58-kb region between simple sequence repeat markers CHR1027 and CHR1030 on the long arm of chromosome 10. The target gene cosegregated with molecular markers CHR1028 and CHR1029. There were five predicted genes in the mapped region. The results from sequencing analysis revealed that there was one base deletion in the first exon of LOC_Os10g26340 encoding cytochrome P450 CYP78A11 in the pla1-5 mutant, which might result in a downstream frame shift and premature termination. These results suggest that the P450 CYP78A11 gene is the candidate gene of PLA1-5.

  6. Resistance to Bacterial Leaf Blight in a Somaclonal Rice Mutant HX-3 at Cellular Level

    Institute of Scientific and Technical Information of China (English)

    GAO Dong-ying; ZHOU Yi-hong; HUANG Xue-qing; SUN Li-hua; LIU Ai-min

    2004-01-01

    The interaction between rice host and its pathogen Xanthomonas oryzae pv. oryzae (Xoo) at cellular level was studied by using a resistant somaclonal mutant HX-3 and its susceptable donor Minghui 63. After inoculation with Xoo strain Zhe 173 (Chinese pathotype Ⅳ), the activity of superoxide dismutase (SOD) and peroxidase (POD) in the callus of Minghui 63 was increased dramatically, and the active oxygen(O2 - .) was produced at a higher rate; Meanwhile, the callus grew slowly with the reduction of protein content. Compared to the activity of SOD and POD, the production rate of O2-. and the fresh weight in HX-3 callus varied little after the inoculation. It could be proposed that there were great differences between the resistance of HX-3 and Mighui 63 at cellular level. There was no difference detected concerning resistance to bacterial leaf blight in HX-3 between the plant and the callus.

  7. Characterization and mapping of a new male sterility mutant of anther advanced dehiscence (t) in rice

    Institute of Scientific and Technical Information of China (English)

    Yi Zhang; Yunfeng Li; Jian Zhang; Fucheng Shen; Yuanxin Huang; Zhiwei Wu

    2008-01-01

    Anther dehiscence is very important for pollen maturation and release.The mutants of anther dehiscence in rice (Oryza sativa L.) arefew,and related research remains poor.A male sterility mutant of anther dehiscence in advance,add(t),has been found in Minghui 63 and its sterility is not sensitive to thermo-photo.To learn the character of sterilization and the function of the add(t) gene,the morphological and cytological studies on the anther and pollen,the ability of the pistil being fertilized,inheritance of the mutant,and mapping of add(t)gene have been conducted.The anther size is normal but the color is white in the mutant against the natural yellow in the wild-type.The pollen is malformed,unstained,and small in the KI-I2 solution.The anther dehiscence is in advance at the bicellular pollen stage.A crossing test indicated that the grain setting ratio of the add(t) is significantly lower than that of the CMS line 2085A.The ability of the pistil being fertilized is most probably decreased by the add(t) gene.The male sterility is controlled by a single recessive gene of add(t).This gene is mapped between the markers of R02004 (InDel) and RM300 (SSR) on chromosome 2,and the genetic distance from the add(t) gene to these markers is 0.78 cM and 4.66 cM,respectively.

  8. Function and characterization of starch synthase I using mutants in rice.

    Science.gov (United States)

    Fujita, Naoko; Yoshida, Mayumi; Asakura, Noriko; Ohdan, Takashi; Miyao, Akio; Hirochika, Hirohiko; Nakamura, Yasunori

    2006-03-01

    Four starch synthase I (SSI)-deficient rice (Oryza sativa) mutant lines were generated using retrotransposon Tos17 insertion. The mutants exhibited different levels of SSI activities and produced significantly lower amounts of SSI protein ranging from 0% to 20% of the wild type. The mutant endosperm amylopectin showed a decrease in chains with degree of polymerization (DP) 8 to 12 and an increase in chains with DP 6 to 7 and DP 16 to 19. The degree of change in amylopectin chain-length distribution was positively correlated with the extent of decrease in SSI activity in the mutants. The structural changes in the amylopectin increased the gelatinization temperature of endosperm starch. Chain-length analysis of amylopectin in the SSI band excised from native-polyacrylamide gel electrophoresis/SS activity staining gel showed that SSI preferentially synthesized DP 7 to 11 chains by elongating DP 4 to 7 short chains of glycogen or amylopectin. These results show that SSI distinctly generates DP 8 to 12 chains from short DP 6 to 7 chains emerging from the branch point in the A or B(1) chain of amylopectin. SSI seemingly functions from the very early through the late stage of endosperm development. Yet, the complete absence of SSI, despite being a major SS isozyme in the developing endosperm, had no effect on the size and shape of seeds and starch granules and the crystallinity of endosperm starch, suggesting that other SS enzymes are probably capable of partly compensating SSI function. In summary, this study strongly suggested that amylopectin chains are synthesized by the coordinated actions of SSI, SSIIa, and SSIIIa isoforms.

  9. Characterization and Genetic Analysis of a Novel Rice Spotted-leaf Mutant HM47 with Broad-spectrum Resistance to Xanthomonas oryzae pv.oryzae(F)

    Institute of Scientific and Technical Information of China (English)

    Bao-Hua Feng; Yang Yang; Yong-Feng Shi; Hai-Chao Shen; Hui-Mei Wang; Qi-Na Huang; Xia Xu

    2013-01-01

    A stable inherited rice spotted-leaf mutant HM47 derived from an EMS-induced IR64 mutant bank was identified.The mutant expressed hypersensitive response (HR)-like symptoms throughout its whole life from the first leaf to the flag leaf,without pathogen invasion.Initiation of the lesions was induced by light under natural summer field conditions.Expression of pathogenesis-related genes including PAL,PO-C1,POX22.3 and PBZ1 was enhanced significantly in association with cell death and accumulation of H2O2 at and around the site of lesions in the mutant in contrast to that in the wild-type (WT).Disease reaction to Xanthomonas oryzae pv.oryzae from the Philippines and China showed that HM47 is a broad-spectrum disease-resistant mutant with enhanced resistance to multiple races of bacterial blight pathogens tested.An F2 progeny test showed that bacterial blight resistance to race HB-17 was cosegregated with the expression of lesions.Genetic analysis indicated that the spotted-leaf trait was controlled by a single recessive gene,tentatively named splHM47,flanked by two insertion/deletion markers in a region of approximately 74 kb on the long arm of chromosome 4.Ten open reading frames are predicted,and all of them are expressed proteins.Isolation and validation of the putative genes are currently underway.

  10. Hygromycin B-induced cell death is partly mediated by reactive oxygen species in rice (Oryza sativa L.).

    Science.gov (United States)

    Oung, Hui-Min; Lin, Ke-Chun; Wu, Tsung-Meng; Chandrika, Nulu Naga Prafulla; Hong, Chwan-Yang

    2015-12-01

    The aminoglycoside antibiotic hygromycin B (Hyg) inhibits prokaryotic, chloroplast and mitochondrial protein synthesis. Because of the toxic effect of Hyg on plant cells, the HPT gene, encoding hygromycin phosphotransferase, has become one of the most widely used selectable markers in plant transformation. Yet the mechanism behind Hyg-induced cell lethality in plants is not clearly understood. In this study, we aimed to decipher this mechanism. With Hyg treatment, rice calli exhibited cell death, and rice seedlings showed severe growth defects, leaf chlorosis and leaf shrinkage. Rice seedlings also exhibited severe lipid peroxidation and protein carbonylation, for oxidative stress damage at the cellular level. The production of reactive oxygen species such as O2(·-), H2O2 and OH(·) was greatly induced in rice seedlings under Hyg stress, and pre-treatment with ascorbate increased resistance to Hyg-induced toxicity indicating the existence of oxidative stress. Overexpression of mitochondrial Alternative oxidase1a gene without HPT selection marker in rice enhanced tolerance to Hyg and attenuated the degradation of protein content, whereas the rice plastidial glutathione reductase 3 mutant showed increased sensitivity to Hyg. These results demonstrate that Hyg-induced cell lethality in rice is not only due to the inhibition of protein synthesis but also mediated by oxidative stress.

  11. Anatomical and Chemical Characteristics of a Rolling Leaf Mutant of Rice and Its Ecophysiological Properties

    Institute of Scientific and Technical Information of China (English)

    BAI Lei; DUAN Zhuang-qin; WANG Jun-min; AN Li-zhe; ZHAO Zhi-guang; CHEN Kun-ming

    2008-01-01

    The anatomical and chemical characteristics of a rolling leaf mutant (r/m) of rice (Oryza sativa L.) and its ecophysiological properties in photosynthesis and apoplastic transport were investigated. Compared with the wild type (WT),the areas of whole vascular bundles and xylem as well as the ratios of xylem area/whole vascular bundles area and xylem area/phloem area were higher in r/m, whereas the area and the width of foliar bulliform cell were lower. The Fourier transform infrared (FTIR) microspectroscopy spectra of foliar cell walls differed greatly between r/m and WT. The r/m exhibited lower protein and polysaccharide contents of foliar cell walls. An obvious reduction of pectin content was also found in r/m by biochemical measurements. Moreover, the rate of photosynthesis was depressed while the conductance of stoma and the intercellular CO2 concentration were enhanced in rim. The PTS fluorescence, which represents the ability of apoplastic transport, was 11% higher in r/m than in WT. These results suggest that the changes in anatomical and chemical characteristics of foliar vascular bundles, such as the reduction of proteins, pectins, and other polysaccharides of foliar cell walls, participate in the leaf rolling mutation, and consequently lead to the reduced photosynthetic dynamics and apoplastic transport ability in the mutant.

  12. Structures of starches from rice mutants deficient in the starch synthase isozyme SSI or SSIIIa.

    Science.gov (United States)

    Hanashiro, Isao; Higuchi, Toshiyuki; Aihara, Satomi; Nakamura, Yasunori; Fujita, Naoko

    2011-05-09

    Amylose and amylopectin of rice mutants deficient in a starch synthase (SS) isozyme in the endosperm, either SSI (ΔSSI) or SSIIIa (ΔSSIIIa), were structurally altered from those of their parent (cv. Nipponbare, Np). The amylose content was higher in the mutants (Np, 15.5%; ΔSSI, 18.2%; ΔSSIIIa, 23.6%), and the molar ratio of branched amylose and its side chains was increased. The chain-length distribution of the β-amylase limit dextrins of amylopectin showed regularity, which appeared consistent with the generally accepted cluster structure, and the degrees of polymerization found at the intersections were taken as the boundaries of the B-chain fractions. The mole % of the B(1)-B(3) fractions was changed slightly in ΔSSI, which is consistent with the proposed role of SSI in elongating the external part of clusters. In ΔSSIIIa, a significant increase in the B(1) fraction and a decrease in the B(2) and B(3) fractions were observed. The internal chain length of the B(2) and B(3) fractions appeared to be slightly altered, suggesting that the deficiency in SS affected the actions of branching enzyme(s).

  13. The α-subunit of the rice heterotrimeric G protein, RGA1, regulates drought tolerance during the vegetative phase in the dwarf rice mutant d1.

    Science.gov (United States)

    Ferrero-Serrano, Ángel; Assmann, Sarah M

    2016-05-01

    Essential in the Green Revolution was the development of high-yielding dwarf varieties of rice (Oryza sativa L.), but their selection was not based on responses to water limitation. We studied physiological responses to progressive drought of the dwarf rice mutant, d1, in which the RGA1 gene, which encodes the GTP-binding α-subunit of the heterotrimeric G protein, is non-functional. Wild-type (WT) plants cease net carbon fixation 11 days after water is withheld, while d1 plants maintain net photosynthesis for an additional week. During drought, d1 plants exhibit greater stomatal conductance than the WT, but both genotypes exhibit the same transpirational water loss per unit leaf area. This is explained by a smaller driving force for water loss in d1 owing to its lower leaf temperatures, consistent with its more erect architecture. As drought becomes more severe, WT plants show an accelerated decline in photosynthesis, which may be exacerbated by the higher leaf temperatures in the WT. We thus show how a rice mutant with dwarf and erect leaves has a decreased susceptibility to water stress. Accordingly, it may be useful to incorporate RGA1 mutation in breeding or biotechnological strategies for development of drought-resistant rice.

  14. Physiological basis and transcriptional profiling of three salt-tolerant mutant lines of rice

    Directory of Open Access Journals (Sweden)

    Concha Domingo

    2016-09-01

    Full Text Available Salinity is a complex trait that affects growth and productivity in many crops, including rice. Mutation induction, a useful tool to generate salt tolerant plants, enables the analysis of plants with similar genetic background, facilitating the understanding of the salt tolerance mechanisms. In this work, we generated three salt tolerant mutant lines by irradiation of a salt-sensitive cultivar plants and screened M2 plants at seedling stage in the presence of high salinity. These three lines, SaT20, SaS62 and SaT58, showed different responses to salinity, but exhibited similar phenotype to wild type plants, except SaT20 that displayed shorter height when grown in the absence of salt. Under salt conditions, all three mutants and the parental line showed similar reduction in yield, although relevant differences in other physiological parameters, such as Na+ accumulation in healthy leaves of SaT20, were registered. Microarray analyses of gene expression profiles in roots revealed the occurrence of common and specific responses in the mutants. The three mutants showed up-regulation of responsive genes, the activation of oxido-reduction process and the inhibition of ion transport. The participation of jasmonate in the plant response to salt was evident by down-regulation of a gene coding for a jasmonate O-methyltransferase. Genes dealing with lipid transport and metabolism were, in general, up-regulated except in SaS62, that also exhibited down-regulation of genes involved in ion transport and Ca2+ signal transduction. The two most tolerant varieties, SaS62 and SaT20, displayed lower levels of transcripts involved in K+ uptake. The physiological study and the description of the expression analysis evidenced that the three lines showed different responses to salt: SaT20 showed a high Na+ content in leaves, SaS62 presented an inhibition of lipid metabolism and ion transport and SaT58 differs in both features in the response to salinity. The analysis of

  15. Physiological Basis and Transcriptional Profiling of Three Salt-Tolerant Mutant Lines of Rice

    Science.gov (United States)

    Domingo, Concha; Lalanne, Eric; Catalá, María M.; Pla, Eva; Reig-Valiente, Juan L.; Talón, Manuel

    2016-01-01

    Salinity is a complex trait that affects growth and productivity in many crops, including rice. Mutation induction, a useful tool to generate salt tolerant plants, enables the analysis of plants with similar genetic background, facilitating the understanding of the salt tolerance mechanisms. In this work, we generated three salt tolerant mutant lines by irradiation of a salt-sensitive cultivar plants and screened M2 plants at seedling stage in the presence of high salinity. These three lines, SaT20, SaS62, and SaT58, showed different responses to salinity, but exhibited similar phenotype to wild type plants, except SaT20 that displayed shorter height when grown in the absence of salt. Under salt conditions, all three mutants and the parental line showed similar reduction in yield, although relevant differences in other physiological parameters, such as Na+ accumulation in healthy leaves of SaT20, were registered. Microarray analyses of gene expression profiles in roots revealed the occurrence of common and specific responses in the mutants. The three mutants showed up-regulation of responsive genes, the activation of oxido-reduction process and the inhibition of ion transport. The participation of jasmonate in the plant response to salt was evident by down-regulation of a gene coding for a jasmonate O-methyltransferase. Genes dealing with lipid transport and metabolism were, in general, up-regulated except in SaS62, that also exhibited down-regulation of genes involved in ion transport and Ca2+ signal transduction. The two most tolerant varieties, SaS62 and SaT20, displayed lower levels of transcripts involved in K+ uptake. The physiological study and the description of the expression analysis evidenced that the three lines showed different responses to salt: SaT20 showed a high Na+ content in leaves, SaS62 presented an inhibition of lipid metabolism and ion transport and SaT58 differs in both features in the response to salinity. The analysis of these salt

  16. Plastidic phosphoglucomutase and ADP-glucose pyrophosphorylase mutants impair starch synthesis in rice pollen grains and cause male sterility.

    Science.gov (United States)

    Lee, Sang-Kyu; Eom, Joon-Seob; Hwang, Seon-Kap; Shin, Dongjin; An, Gynheung; Okita, Thomas W; Jeon, Jong-Seong

    2016-10-01

    To elucidate the starch synthesis pathway and the role of this reserve in rice pollen, we characterized mutations in the plastidic phosphoglucomutase, OspPGM, and the plastidic large subunit of ADP-glucose (ADP-Glc) pyrophosphorylase, OsAGPL4 Both genes were up-regulated in maturing pollen, a stage when starch begins to accumulate. Progeny analysis of self-pollinated heterozygous lines carrying the OspPGM mutant alleles, osppgm-1 and osppgm-2, or the OsAGPL4 mutant allele, osagpl4-1, as well as reciprocal crosses between the wild type (WT) and heterozygotes revealed that loss of OspPGM or OsAGPL4 caused male sterility, with the former condition rescued by the introduction of the WT OspPGM gene. While iodine staining and transmission electron microscopy analyses of pollen grains from homozygous osppgm-1 lines produced by anther culture confirmed the starch null phenotype, pollen from homozygous osagpl4 mutant lines, osagpl4-2 and osagpl4-3, generated by the CRISPR/Cas system, accumulated small amounts of starch which were sufficient to produce viable seed. Such osagpl4 mutant pollen, however, was unable to compete against WT pollen successfully, validating the important role of this reserve in fertilization. Our results demonstrate that starch is mainly polymerized from ADP-Glc synthesized from plastidic hexose phosphates in rice pollen and that starch is an essential requirement for successful fertilization in rice.

  17. Food protein induced enterocolitis syndrome caused by rice beverage.

    Science.gov (United States)

    Caminiti, Lucia; Salzano, Giuseppina; Crisafulli, Giuseppe; Porcaro, Federica; Pajno, Giovanni Battista

    2013-05-14

    Food protein-induced enterocolitis syndrome (FPIES) is an uncommon and potentially severe non IgE-mediated gastrointestinal food allergy. It is usually caused by cow's milk or soy proteins, but may also be triggered by ingestion of solid foods. The diagnosis is made on the basis of clinical history and symptoms. Management of acute phase requires fluid resuscitation and intravenous steroids administration, but avoidance of offending foods is the only effective therapeutic option.Infant with FPIES presented to our emergency department with vomiting, watery stools, hypothension and metabolic acidosis after ingestion of rice beverage. Intravenous fluids and steroids were administered with good clinical response. Subsequently, a double blind placebo control food challenge (DBPCFC) was performed using rice beverage and hydrolyzed formula (eHF) as placebo. The "rice based formula" induced emesis, diarrhoea and lethargy. Laboratory investigations reveal an increase of absolute count of neutrophils and the presence of faecal eosinophils. The patient was treated with both intravenous hydration and steroids. According to Powell criteria, oral food challenge was considered positive and diagnosis of FPIES induced by rice beverage was made. Patient was discharged at home with the indication to avoid rice and any rice beverage as well as to reintroduce hydrolyzed formula. A case of FPIES induced by rice beverage has never been reported. The present case clearly shows that also beverage containing rice proteins can be responsible of FPIES. For this reason, the use of rice beverage as cow's milk substitute for the treatment of non IgE-mediated food allergy should be avoided.

  18. New Rice Mutants(NR676 and NR827)Low in Nitrate Reduction

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Mutants deficient in nitrate reductase(NR) were classfied into two groups,nia types are deficient in apoprotein and cnx types are lack of Mo- Co. This paper reports the characteristics of low NR mutants,which are not deficient in NR activity but 40%~60% of the NR activity of the wild type,Nipponbare. Mutants NR676 and NR827 were selected as seedings showing poor growth with nitrate as sole mitrogen source from M2 population. They exhibited chlorate resistance. Genetic analysis in the F1 and F2 indicated that chlorate resistance in the mutants was transmitted by a single recessive nuclear gene and that NR676 and NR827 were induced by a mutation at a single locus. In the F2 population,all of the seedings showing yellow green leaves were resistant to chlorate and low in NR activity. Photosynthetic rate and mRNA expression of NR676 and NR827 were lower than wild type. These results suggested that low NR activity and chlorate resistance of NR676 and NR827 were caused by a defect in photosynthetic process.

  19. Increased leaf photosynthesis caused by elevated stomatal conductance in a rice mutant deficient in SLAC1, a guard cell anion channel protein

    OpenAIRE

    2012-01-01

    In rice (Oryza sativa L.), leaf photosynthesis is known to be highly correlated with stomatal conductance; however, it remains unclear whether stomatal conductance dominantly limits the photosynthetic rate. SLAC1 is a stomatal anion channel protein controlling stomatal closure in response to environmental [CO2]. In order to examine stomatal limitations to photosynthesis, a SLAC1-deficient mutant of rice was isolated and characterized. A TILLING screen of N-methyl-N-nitrosourea-derived mutant ...

  20. Genetic analysis and fine-mapping of a dwarfing with withered leaf-tip mutant in rice

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    A dwarf mutant of rice(Oryza.sativa L.)by mutagenesis of ethylene methylsulfonate(EMS)treatment from Nipponbare was identified.The mutant exhibited phenotypes of dwarfism and withered leaf tip(dwll).Based on the internode length of dwl1,this mutant be longs to the dm type of dwarfing.Analysis of elongation of the second sheath and α-amylase activity in endosperm showed that the phenotype caused by dwll was insensitive to gibberellin acid treatment.Using a large F2 population derived from a cross between the dwll and an indica rice variety,TN1,the DWLl gene was mapped to the terminal region of the long arm of chromosome 3.Fine-mapping de-limited it into a 46 kb physical distance between two STS markers,HL921 and HL944,where 6 open reading frames were predicted.Cloning of DWL1 will contribute to dissecting molecular mechanism that regulates plant height in rice,which will be beneficial to molecular assisted selection of this important trait.

  1. Characterization of the endosperm starch and the pleiotropic effects of biosynthetic enzymes on their properties in novel mutant rice lines with high resistant starch and amylose content.

    Science.gov (United States)

    Itoh, Yuuki; Crofts, Naoko; Abe, Misato; Hosaka, Yuko; Fujita, Naoko

    2017-05-01

    Resistant starch (RS) is beneficial to human health. In order to reduce the current prevalence of diabetes and obesity, several transgenic and mutant crops containing high RS content are being developed. RS content of steamed rice with starch-branching enzyme (BE)IIb-deficient mutant endosperms is considerably high. To understand the mechanisms of RS synthesis and to increase RS content, we developed novel mutant rice lines by introducing the gene encoding starch synthase (SS)IIa and/or granule-bound starch synthase (GBSS)I from an indica rice cultivar into a japonica rice-based BEIIb-deficient mutant line, be2b. Introduction of SSIIa from an indica rice cultivar produced higher levels of amylopectin chains with degree of polymerization (DP) 11-18 than those in be2b; the extent of the change was slight due to the shortage of donor chains for SSIIa (DP 6-12) owing to BEIIb deficiency. The introduction of GBSSI from an indica rice cultivar significantly increased amylose content (by approximately 10%) in the endosperm starch. RS content of the new mutant lines was the same as or slightly higher than that of the be2b parent line. The relationship linking starch structure, RS content, and starch biosynthetic enzymes in the new mutant lines has also been discussed. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Identification of rice ethylene-response mutants and characterization of MHZ7/OsEIN2 in distinct ethylene response and yield trait regulation.

    Science.gov (United States)

    Ma, Biao; He, Si-Jie; Duan, Kai-Xuan; Yin, Cui-Cui; Chen, Hui; Yang, Chao; Xiong, Qing; Song, Qing-Xin; Lu, Xiang; Chen, Hao-Wei; Zhang, Wan-Ke; Lu, Tie-Gang; Chen, Shou-Yi; Zhang, Jin-Song

    2013-11-01

    Ethylene plays essential roles in adaptive growth of rice plants in water-saturating environment; however, ethylene signaling pathway in rice is largely unclear. In this study, we report identification and characterization of ethylene-response mutants based on the specific ethylene-response phenotypes of etiolated rice seedlings, including ethylene-inhibited root growth and ethylene-promoted coleoptile elongation, which is different from the ethylene triple-response phenotype in Arabidopsis. We establish an efficient system for screening and a set of rice mutants have been identified. Genetic analysis reveals that these mutants form eight complementation groups. All the mutants show insensitivity or reduced sensitivity to ethylene in root growth but exhibit differential responses in coleoptile growth. One mutant group mhz7 has insensitivity to ethylene in both root and coleoptile growth. We identified the corresponding gene by a map-based cloning method. MHZ7 encodes a membrane protein homologous to EIN2, a central component of ethylene signaling in Arabidopsis. Upon ethylene treatment, etiolated MHZ7-overexpressing seedlings exhibit enhanced coleoptile elongation, increased mesocotyl growth and extremely twisted short roots, featuring enhanced ethylene-response phenotypes in rice. Grain length was promoted in MHZ7-transgenic plants and 1000-grain weight was reduced in mhz7 mutants. Leaf senescent process was also affected by MHZ7 expression. Manipulation of ethylene signaling may improve adaptive growth and yield-related traits in rice.

  3. Construction of chimeric inducible promoters by elicitors of rice fungal blast pathogen and their expression in transgenic rice

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    The promoter fragments of wheat GstA1 and potato Gst1 have been amplified by PCR, cloned and fused respectively to the minimal promoter sequence of rice actin gene (Act1)) and its 5′ untranslated leader sequence together with GUS. The constructs with 2 chimeric promoters (WGA and PGA) have been transferred into rice in order to analyze their inducibility patterns in transgenic rice plants. The results show that: WGA and PGA are both inducible by elicitors of Pyricularia oryzae in transgenic rice cells; the intronⅠ of rice Act1 gene is important for the heterogenic expression of monocot and dicot promoter elements in rice; and the Act1 minimal promoter and its 5′untranslated leader sequence produced low level background expression in rice.

  4. Biomass digestibility is predominantly affected by three factors of wall polymer features distinctive in wheat accessions and rice mutants

    Science.gov (United States)

    2013-01-01

    Background Wheat and rice are important food crops with enormous biomass residues for biofuels. However, lignocellulosic recalcitrance becomes a crucial factor on biomass process. Plant cell walls greatly determine biomass recalcitrance, thus it is essential to identify their key factors on lignocellulose saccharification. Despite it has been reported about cell wall factors on biomass digestions, little is known in wheat and rice. In this study, we analyzed nine typical pairs of wheat and rice samples that exhibited distinct cell wall compositions, and identified three major factors of wall polymer features that affected biomass digestibility. Results Based on cell wall compositions, ten wheat accessions and three rice mutants were classified into three distinct groups each with three typical pairs. In terms of group I that displayed single wall polymer alternations in wheat, we found that three wall polymer levels (cellulose, hemicelluloses and lignin) each had a negative effect on biomass digestibility at similar rates under pretreatments of NaOH and H2SO4 with three concentrations. However, analysis of six pairs of wheat and rice samples in groups II and III that each exhibited a similar cell wall composition, indicated that three wall polymer levels were not the major factors on biomass saccharification. Furthermore, in-depth detection of the wall polymer features distinctive in rice mutants, demonstrated that biomass digestibility was remarkably affected either negatively by cellulose crystallinity (CrI) of raw biomass materials, or positively by both Ara substitution degree of non-KOH-extractable hemicelluloses (reverse Xyl/Ara) and p-coumaryl alcohol relative proportion of KOH-extractable lignin (H/G). Correlation analysis indicated that Ara substitution degree and H/G ratio negatively affected cellulose crystallinity for high biomass enzymatic digestion. It was also suggested to determine whether Ara and H monomer have an interlinking with cellulose chains

  5. Fermentation and quality of yellow pigments from golden brown rice solid culture by a selected Monascus mutant.

    Science.gov (United States)

    Yongsmith, Busaba; Thongpradis, Panida; Klinsupa, Worawan; Chantrapornchai, Withida; Haruthaithanasan, Vichai

    2013-10-01

    A single peak (λmax 370) yellow pigment-producing mutant derived from Monascus sp. TISTR 3179 was used for the pigment production in solid rice culture. Various factors affecting yellow tones were investigated. Hom-mali rice variety was the best amongst five Thai local varieties used for fungus culture. It was also better than corn, mungbean, soybean, potato, sweet potato, or cassava tubers. The moisture content and temperature were the key environmental factors affecting the color tones of creamy, tangerine, and golden brown rice solid cultures. The golden brown rice culture gave the highest yellow pigment concentration. Under an optimum room temperature of 28-32 °C, an initial moisture content of 42 %, and 7-day-old inoculum size of 2 % (v/w) the maximum yield at 2,224.63 A370U/gdw of yellow pigment was produced. A mellow yellow powder at 550 A370U/gdw could be obtained using spray-drying techniques. The powder had a moisture content of 5.15 %, a water activity value of 0.398, a hue angle of 73.70 ° (yellowish orange), high lightness (L) of 74.63, color saturation (C) of 28.97, a neutral pH of 7.42, 0.12 % acidity and solubility of 0.211 g/10 ml. It was noteworthy that the Chinese fresh noodle with spray-dried yellow powder showed no discoloration during 8-day storage.

  6. Analysis of Salicylic Acid Induced Proteins in Rice

    Institute of Scientific and Technical Information of China (English)

    1999-01-01

    An analysis using SDS-PAGE of acidic and basic protein fractions extracted from rice seedling treated with salicylic acid (SA) yielded several new proteins, some of which are similar in relative molecular mass to PR-1a,c, PR-2, 2e and PR-3d, 3e of tobacco.Direct assays for peroxidases and β-1,3-glucanases demonstrated that the activities of the two enzymes in the rice seedlings increased rapidly with time after SA treatment, reaching a maximum 6 days after treatment.Disease resistance tests showed that SA treated rice seedlings stunted the development of blight lesions and displayed higher resistance to rice blight pathogen (Xanthomonas oryzea pv.oryzea).The data suggest that the treatment with SA, even for plants with high endogenous SA levels such as rice, may induce the appearance of new proteins and the formation of disease resistance.The results contribute to the analysis of the SA role in rice systemic acquired resistance.

  7. Phosphine-induced physiological and biochemical responses in rice seedlings.

    Science.gov (United States)

    Mi, Lina; Niu, Xiaojun; Lu, Meiqing; Ma, Jinling; Wu, Jiandong; Zhou, Xingqiu

    2014-04-01

    Paddy fields have been demonstrated to be one of the major resources of atmospheric phosphine and may have both positive and negative effects on rice plants. To elucidate the physiological and biochemical responses of rice plants to phosphine, rice seedlings (30 d old) were selected as a model plant and were treated with different concentrations of phosphine (0, 1.4, 4.2, and 7.0 mg m(-3)). Antioxidant enzymes, including superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), and lipid peroxidation measured via malondialdehyde (MDA) were determined as indicators of the physiological and biochemical responses of the rice seedlings to phosphine exposure. Increasing concentrations of phosphine treatment enhanced the activity of SOD, POD, and CAT. In addition, the MDA content increased with increasing concentrations of phosphine. These results suggested that antioxidant enzymes played important roles in protecting rice seedlings from ROS damage. Moreover, rice seedlings were able to cope with the oxidative stress induced by low concentrations of phosphine via an increase in antioxidant enzymatic activities. However, oxidative stress may not fully be prevented when the plants were exposed to higher concentrations of phosphine.

  8. Elicitor induced activation of the methylerythritol phosphate pathway toward phytoalexins biosynthesis in rice.

    Science.gov (United States)

    Okada, Atsushi; Shimizu, Takafumi; Okada, Kazunori; Kuzuyama, Tomohisa; Koga, Jinichiro; Shibuya, Naoto; Nojiri, Hideaki; Yamane, Hisakazu

    2007-09-01

    Diterpenoid phytoalexins such as momilactones and phytocassanes are produced via geranylgeranyl diphosphate in suspension-cultured rice cells after treatment with a chitin elicitor. We have previously shown that the production of diterpene hydrocarbons leading to phytoalexins and the expression of related biosynthetic genes are activated in suspension-cultured rice cells upon elicitor treatment. To better understand the elicitor-induced activation of phytoalexin biosynthesis, we conducted microarray analysis using suspension-cultured rice cells collected at various times after treatment with chitin elicitor. Hierarchical cluster analysis revealed two types of early-induced expression (EIE-1, EIE-2) nodes and a late-induced expression (LIE) node that includes genes involved in phytoalexins biosynthesis. The LIE node contains genes that may be responsible for the methylerythritol phosphate (MEP) pathway, a plastidic biosynthetic pathway for isopentenyl diphosphate, an early precursor of phytoalexins. The elicitor-induced expression of these putative MEP pathway genes was confirmed by quantitative reverse-transcription PCR. 1-Deoxy-D: -xylulose 5-phosphate synthase (DXS), 1-deoxy-D: -xylulose 5-phosphate reductoisomerase (DXR), and 4-(cytidine 5'-diphospho)-2-C-methyl-D: -erythritol synthase (CMS), which catalyze the first three committed steps in the MEP pathway, were further shown to have enzymatic activities that complement the growth of E. coli mutants disrupted in the corresponding genes. Application of ketoclomazone and fosmidomycin, inhibitors of DXS and DXR, respectively, repressed the accumulation of diterpene-type phytoalexins in suspension cells treated with chitin elicitor. These results suggest that activation of the MEP pathway is required to supply sufficient terpenoid precursors for the production of phytoalexins in infected rice plants.

  9. How does the NPM1 mutant induce leukemia?

    Directory of Open Access Journals (Sweden)

    Paolo Sportoletti

    2011-06-01

    Full Text Available NPM1 is the most frequently mutated gene in AML and the role of the NPM1 mutant in acute myeloid leukemia along with its leukemogenic potential are still under investigation. NPM1 genetic alterations can contribute to leukemogenesis through the direct oncogenic effect of the mutant protein and the concomitant loss of one functional allele. Npm1 loss determines tumor development in the mouse while in human NPM1 maps in a chromosomal region frequently loss in myelodysplastic syndrome (MDS. The NPM1 mutant cytoplasmic delocalization in leukemic blasts alters multiple cellular pathways through either loss or gain of function effects on different protein partners. Here we discuss the most relevant studies on the role of the NPM1 molecule in hematological malignancies and both in vitro and in vivo studies that are trying to elucidate the way by which the NPM1 mutation induces leukemia.

  10. Genetic analysis and gene fine mapping for a rice novel mutant (rl9(t)) with rolling leaf character

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Leaf shape is an important parameter for ideotype breeding in rice, and the rolling of leaf is also beneficial to efficient ripening of grains. This encourages the explorations of new genes that regulate leaf shape. In this study, genetic analysis and gene mapping were carried out for a novel rolling leaf mutant identified from japonica variety Zhonghua 11. The SSR marker analysis showed that the mutant was controlled by a single recessive gene (rl9(t)) located on chromosome 9. Fine mapping of the Rl9(t) locus was conducted with 30 new STS markers developed around Rl9(t) anchored region based on the sequence diversity between Nipponbare and 93-11. The fine mapping necessitated the contruction of a PAC contig encompassing the Rl9(t) locus, which was delimited to a 42 kb region. This could therefore enhance the cloning of the target gene in further studies.

  11. Cloning and Expression of Gene Responsible for High-Tillering Dwarf Phenotype in Indica Rice Mutant gsor23

    Directory of Open Access Journals (Sweden)

    Shou-jiang YUAN

    2013-09-01

    Full Text Available High-tillering dwarf mutant gsor23 was generated from an indica rice variety Indica9 radiatied by γ-ray. Genetic analysis showed that this phenotype was controlled by one single recessive gene, which was mapped within a physical distance of 386 kb between two insertion-deletion (InDel markers C1-WT2 and C1-WT4 on the long arm of chromosome 1. There is a known gene D10 within this region, the mutation of which causes high-tillering in rice. Sequence analysis of the D10 allele in gsor23 revealed that the base cytosine (C at the 404th position in the coding region was deleted, which would cause frameshift mutation after the 134th amino acids. The mutation site and indica background of gsor23 were different from the previously reported japonica mutants d10-1 and d10-2. Therefore, gsor23 is a novel allelic mutant of D10 which encodes the carotenoid-cleaving dioxygenase 8 (CCD8, a key enzyme involved in the biosynthesis of the new plant hormone strigolactones (SLs. After treatment with GR24, a synthetic analogue of SLs, the high-tillering phenotype of gsor23 was restored to normal. Real-time RT-PCR analysis showed that D10 expression was high in roots, but low in leaves. Compared with the wild type Indica9, the expression of the SL biosynthesis gene D10 was upregulated, while genes likely involved in the SL signal transduction pathway such as D3 and D14 were down-regulated in the gsor23 mutant.

  12. Isolation and characterization of Escherichia coli mutants lacking inducible cyanase.

    Science.gov (United States)

    Guilloton, M; Karst, F

    1987-03-01

    To determine the physiological role of cyanate aminohydrolase (cyanase, EC 3.5.5.3) in bacteria, mutants of Escherichia coli K12 devoid of this inducible activity were isolated and their properties investigated. Five independent mutations were localized next to lac; three of them lay between lacY and codA. Thus cyanase activity could depend on the integrity of one gene or set of clustered genes; we propose for this locus the symbol cnt. Growth of the mutant stains was more sensitive to cyanate than growth of wild-type strains. This difference was noticeable in synthetic medium in the presence of low concentrations of cyanate (less than or equal to 1 mM). Higher concentrations inhibited growth of both wild-type and mutant strains. Urea in aqueous solutions dissociates slowly into ammonium cyanate. Accordingly wild-type strains were able to grow on a synthetic medium containing 0.5 M-urea whereas mutants lacking cyanase were not. We conclude that cyanase could play a role in destroying exogenous cyanate originating from the dissociation of carbamoyl compounds such as urea; alternatively cyanate might constitute a convenient nitrogen source for bacteria able to synthesize cyanase in an inducible way.

  13. Genetic analysis and gene mapping of a new rolled-leaf mutant in rice (Oryza sativa L.)

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    To understand the development of rice leaf blades,we identified a new rolled-leaf mutant,w32,from indica cultivar IR64 through EMS mutagenesis. The mutant showed a stable rolled-leaf phenotype throughout the life cycle. Two F2 populations were developed by crossing w32 to cultivar IR24 and PA64. Genetic analysis showed that the rolled-leaf phenotype was controlled by a single recessive gene. To determine the location of the gene,bulked segregant analysis was carried out using mutant and wild-type DNA pools and 1846 mutant-type F2 individuals derived from the cross w32/PA64 were genotyped to locate the gene on the short arm of chromosome 7. The rolled-leaf gene,tentatively named rl11(t),is likely a new gene as no other rolled-leaf genes have been identified near the region. By developing new SSR and InDel markers,the gene was delimited to a 52 kb region near the end of the short chromosome arm. Further fine mapping and cloning of the gene are currently underway.

  14. Altered disease development in the eui mutants and Eui overexpressors indicates that gibberellins negatively regulate rice basal disease resistance.

    Science.gov (United States)

    Yang, Dong-Lei; Li, Qun; Deng, Yi-Wen; Lou, Yong-Gen; Wang, Mu-Yang; Zhou, Guo-Xing; Zhang, Ying-Ying; He, Zu-Hua

    2008-05-01

    Gibberellins (GAs) form a group of important plant tetracyclic diterpenoid hormones that are involved in many aspects of plant growth and development. Emerging evidence implicates that GAs also play roles in stress responses. However, the role of GAs in biotic stress is largely unknown. Here, we report that knockout or overexpression of the Elongated uppermost internode (Eui) gene encoding a GA deactivating enzyme compromises or increases, respectively, disease resistance to bacterial blight (Xanthomonas oryzae pv. oyrzae) and rice blast (Magnaporthe oryzae). Exogenous application of GA(3) and the inhibitor of GA synthesis (uniconazol) could increase disease susceptibility and resistance, respectively, to bacterial blight. Similarly, uniconazol restored disease resistance of the eui mutant and GA(3) decreased disease resistance of the Eui overexpressors to bacterial blight. Therefore, the change of resistance attributes to GA levels. In consistency with this, the GA metabolism genes OsGA20ox2 and OsGA2ox1 were down-regulated during pathogen challenge. We also found that PR1a induction was enhanced but the SA level was decreased in the Eui overexpressor, while the JA level was reduced in the eui mutant. Together, our current study indicates that GAs play a negative role in rice basal disease resistance, with EUI as a positive modulator through regulating the level of bioactive GAs.

  15. Altered Disease Development in the eui Mutants and Eui Overexpressors Indicates that Gibberellins Negatively Regulate Rice Basal Disease Resistance

    Institute of Scientific and Technical Information of China (English)

    Dong-Lei Yang; Qun Li; Yi-Wen Deng; Yong-Gen Lou; Mu-Yang Wang; Guo-Xing Zhou; Ying-Ying Zhang; Zu-Hua He

    2008-01-01

    Gibberellins (GAs) form a group of important plant tetracyclic diterpenoid hormones that are involved in many aspects of plant growth and development. Emerging evidence implicates that GAs also play roles in stress responses. However, the role of GAs in biotic stress is largely unknown. Here, we report that knockout or overexpression of the Elongated uppermost internode (Eui) gene encoding a GA deactivating enzyme compromises or increases, respectively, disease resistance to bacterial blight (Xanthomonas oryzae pv. oyrzae) and rice blast (Magnaporthe oryzae). Exogenous application of GA and the inhibitor of GA synthesis (uniconazol) could increase disease susceptibility and resistance, respectively, to bacterial blight. Similarly, uniconazol restored disease resistance of the eui mutant and GA3 decreased disease resistance of the Eui overexpressors to bacterial blight. Therefore, the change of resistance attributes to GA levels. In consistency with this, the GA metabolism genes OsGA2Oox2 and OsGA2oxl were down-regulated during pathogen challenge. We also found that PR1a induction was enhanced but the SA level was decreased in the Eui overexpressor, while the JA level was reduced in the eui mutant. Together, our current study indicates that GAs play a negative role in rice basal disease resistance, with EUI as a positive modulator through regulating the level of bioactive GAs.

  16. Radiation induced useful mutants of Japanese mint (Mentha arvesis L. )

    Energy Technology Data Exchange (ETDEWEB)

    Kak, S.N.; Kaul, B.L.

    1980-01-01

    With a view to inducing mutations, dormant suckers of Mentha arvensis var. piperascens were exposed to X- and gamma rays. Individual clones raised from these treatments were screened for morphological and biochemical traits. Clones with apparent mutations in the first vegetative generation were propagated through second generation and those breeding true for the mutated character were isolated. In this way several mutants were isolated, some of which were found to be novel with respect to the quality of oil, besides those economically useful. The study shows that induced mutations could be very useful in the breeding of vegetatively propagated plants.

  17. Biomolecular analyses of starch and starch granule proteins in the high-amylose rice mutant Goami 2.

    Science.gov (United States)

    Butardo, Vito M; Daygon, Venea Dara; Colgrave, Michelle L; Campbell, Peter M; Resurreccion, Adoracion; Cuevas, Rosa Paula; Jobling, Stephen A; Tetlow, Ian; Rahman, Sadequr; Morell, Matthew; Fitzgerald, Melissa

    2012-11-21

    Elevated proportions of amylose in cereals are commonly associated with either the loss of starch branching or starch synthase activity. Goami 2 is a high-amylose mutant of the temperate japonica rice variety Ilpumbyeo. Genotyping revealed that Goami 2 and Ilpumbyeo carry the same alleles for starch synthase IIa and granule-bound starch synthase I genes. Analyses of granule-bound proteins revealed that SSI and SSIIa accumulate inside the mature starch granules of Goami 2, which is similar to the amylose extender mutant IR36ae. However, unlike the amylose extender mutants, SBEIIb was still detectable inside the starch granules of Goami 2. Detection of SBEIIb after protein fractionation revealed that most of the SBEIIb in Goami 2 accumulates inside the starch granules, whereas most of it accumulates at the granule surface in Ilpumbyeo. Exhaustive mass spectrometric characterisations of granule-bound proteins failed to detect any peptide sequence mutation or major post-translational modifications in Goami 2. Moreover, the signal peptide was found to be cleaved normally from the precursor protein, and there is no apparent N-linked glycosylation. Finally, no difference was found in the SBEIIb structural gene sequence of Goami 2 compared with Ilpumbyeo. In contrast, a G-to-A mutation was detected in the SBEIIb gene of IR36ae located at the splice site between exon and intron 11, which could potentially introduce a premature stop codon and produce a truncated form of SBEIIb. It is suggested that the mutation responsible for producing high amylose in Goami 2 is not due to a defect in SBEIIb gene as was observed in IR36ae, even though it produces a phenotype analogous to the amylose extender mutation. Understanding the molecular genetic basis of this mutation will be important in identifying novel targets for increasing amylose and resistant starch contents in rice and other cereals.

  18. Role of ethylene signaling in the production of rice volatiles induced by the rice brown planthopper Nilaparvata lugens

    Institute of Scientific and Technical Information of China (English)

    LU Yujie; WANG Xia; LOU Yonggen; CHENG Jiaan

    2006-01-01

    Ethylene signaling pathway plays an important role in induced plant direct defense against herbivores and pathogens; however, up to now, only few researches have focused on its role in induced plant indirect defense, i.e. the release of herbivore-induced volatiles, and the results are variable.Using a model system consisting of rice plants, the rice brown planthopper Nilaparvata lugens and its egg parasitoid Anagrus nilaparvatae, we examined the role of ethylene signaling in the production of rice volatiles induced by N. lugens by measuring both the timing of herbivore-induced ethylene levels and the relationships between ethylene, rice volatiles and attraction of the parasitoid. N. lugens infestation significantly enhanced the release of ethylene during 2-24 h after infestation. Plants treated with ethephon, a compound that breaks down to release ethylene at cytoplasmic pH, released volatiles profiles similar to those released by N.lugens-infested plants,and both of them showed an equal attraction of the parasitoid. Moreover, pretreatment with 1-MCP, an inhibitor of ethylene perception, reduced the release of most of rice volatiles whose amount was enhanced by N.lugens infestation and decreased the attractiveness to the parasitoid. These results demonstrate that ethylene signaling is required for the production of rice volatiles induced by N.lugens.

  19. GPNMB ameliorates mutant TDP-43-induced motor neuron cell death.

    Science.gov (United States)

    Nagahara, Yuki; Shimazawa, Masamitsu; Ohuchi, Kazuki; Ito, Junko; Takahashi, Hitoshi; Tsuruma, Kazuhiro; Kakita, Akiyoshi; Hara, Hideaki

    2017-08-01

    Glycoprotein nonmetastatic melanoma protein B (GPNMB) aggregates are observed in the spinal cord of amyotrophic lateral sclerosis (ALS) patients, but the detailed localization is still unclear. Mutations of transactive response DNA binding protein 43kDa (TDP-43) are associated with neurodegenerative diseases including ALS. In this study, we evaluated the localization of GPNMB aggregates in the spinal cord of ALS patients and the effect of GPNMB against mutant TDP-43 induced motor neuron cell death. GPNMB aggregates were not localized in the glial fibrillary acidic protein (GFAP)-positive astrocyte and ionized calcium binding adaptor molecule-1 (Iba1)-positive microglia. GPNMB aggregates were localized in the microtubule-associated protein 2 (MAP-2)-positive neuron and neurofilament H non-phosphorylated (SMI-32)-positive neuron, and these were co-localized with TDP-43 aggregates in the spinal cord of ALS patients. Mock or TDP-43 (WT, M337V, and A315T) plasmids were transfected into mouse motor neuron cells (NSC34). The expression level of GPNMB was increased by transfection of mutant TDP-43 plasmids. Recombinant GPNMB ameliorated motor neuron cell death induced by transfection of mutant TDP-43 plasmids and serum-free stress. Furthermore, the expression of phosphorylated ERK1/2 and phosphorylated Akt were decreased by this stress, and these expressions were increased by recombinant GPNMB. These results indicate that GPNMB has protective effects against mutant TDP-43 stress via activating the ERK1/2 and Akt pathways, and GPNMB may be a therapeutic target for TDP-43 proteinopathy in familial and sporadic ALS. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  20. Brewers’ Rice: A By-Product from Rice Processing Provides Natural Hepatorenal Protection in Azoxymethane-Induced Oxidative Stress in Rats

    Directory of Open Access Journals (Sweden)

    Bee Ling Tan

    2015-01-01

    Full Text Available Brewers’ rice, which is known locally as temukut, is a mixture of broken rice, rice bran, and rice germ. Our present study was designed to identify the effect of brewers’ rice on the attenuation of liver and kidney damage induced by azoxymethane (AOM. Alanine transaminase (ALT, alkaline phosphatase (ALP, aspartate transaminase (AST, creatinine, and urea were evaluated to understand potential hepatoprotective effects and the ability of brewers’ rice to attenuate kidney pathology induced by AOM treatment. Liver and kidney tissues were evaluated by hematoxylin and eosin (H&E staining. Overall analyses revealed that brewers’ rice improved the levels of serum markers in a manner associated with better histopathological outcomes, which indicated that brewers’ rice could enhance recovery from hepatocyte and kidney damage. Taken together, these results suggest that brewers’ rice could be used in future applications to combat liver and kidney disease.

  1. Nitric oxide ameliorates zinc oxide nanoparticles-induced phytotoxicity in rice seedlings.

    Science.gov (United States)

    Chen, Juan; Liu, Xiang; Wang, Chao; Yin, Shan-Shan; Li, Xiu-Ling; Hu, Wen-Jun; Simon, Martin; Shen, Zhi-Jun; Xiao, Qiang; Chu, Cheng-Cai; Peng, Xin-Xiang; Zheng, Hai-Lei

    2015-10-30

    Nitric oxide (NO) has been found to function in enhancing plant tolerance to various environmental stresses. However, role of NO in relieving zinc oxide nanoparticles (ZnO NPs)-induced phytotoxicity remains unknown. Here, sodium nitroprusside (SNP, a NO donor) was used to investigate the possible roles and the regulatory mechanisms of NO in counteracting ZnO NPs toxicity in rice seedlings. Our results showed that 10 μM SNP significantly inhibited the appearance of ZnO NP toxicity symptoms. SNP addition significantly reduced Zn accumulation, reactive oxygen species production and lipid peroxidation caused by ZnO NPs. The protective role of SNP in reducing ZnO NPs-induced oxidative damage is closely related to NO-mediated antioxidant system. A decrease in superoxide dismutase activity, as well as an increase in reduced glutathione content and peroxidase, catalase and ascorbate peroxidase activity was observed under SNP and ZnO NPs combined treatments, compared to ZnO NPs treatment alone. The relative transcript abundance of corresponding antioxidant genes exhibited a similar change. The role of NO in enhancing ZnO NPs tolerance was further confirmed by genetic analysis using a NO excess mutant (noe1) and an OsNOA1-silenced plant (noa1) of rice. Together, this study provides the first evidence indicating that NO functions in ameliorating ZnO NPs-induced phytotoxicity.

  2. Changes in Thermostability of Photosystem Ⅱ and Leaf Lipid Composition of Rice Mutant with Deficiency of Light-harvesting Chlorophyll Protein Complexes

    Institute of Scientific and Technical Information of China (English)

    Yunlai Tang; Mei Chen; Yinong Xu; Tingyun Kuang

    2007-01-01

    We studied the difference in thermostability of photosystem Ⅱ (PSⅡ) and leaf lipid composition between a T-DNA insertion mutant rice (Oryza sativa L.) VG28 and its wild type Zhonghua11. Native green gel and SDS-PAGE electrophoreses revealed that the mutant VG28 lacked all light-harvesting chlorophyll a/b protein complexes. Both the mutant and wild type were sensitive to high temperatures, and the maximal efficiency of PSⅡ photochemistry (Fv/Fm) and oxygen-evolving activity of PSⅡ in leaves significantly decreased with increasing temperature. However, the PSⅡ activity of the mutant was markedly more sensitive to high temperatures than that of the wild type. Lipid composition analysis showed that the mutant had less phosphatidylglycerol and sulfoquinovosyl diacylglycerol compared with the wild type. Fatty acid analysis revealed that the mutant had an obvious decrease in the content of unsaturation of membrane lipids on the thermostability of PSll are discussed.

  3. Recovery and Biological Properties of Nitrate Non-utilizing Mutants of Rice Blast, Magnaporthe grisea

    Institute of Scientific and Technical Information of China (English)

    ZHANG Chuan-qing; ZHOU Ming-guo

    2004-01-01

    Eleven nitrate non-utilizing (nit) mutants were recovered from six isolates of Magnaporthe grisea cultured on MM media amended with 60 g/L potassium chlorate, with a frequency of 1.42 %. Some biological properties, such as growth rate, growth biomass, cultural characters, conidial production, sexual reproduction ability, and pathogenicity were compared between nit mutants and their parent isolates. Results showed that all the nit mutants were resistant to chlorate. Some important biological properties such as the growth rate on YPSA, conidial production ability on TPSA, pathogenicity, had no significant differences between nit mutants and their parent isolates. Mating type didn't change, but perithecia production ability of fertile isolates changed significantly as compared with that of their parent isolates. Therefore, the nit can be used as a genetic marker to study the genetics such as pathogenicity, fungicide resistance in Magnaporthe grisea.

  4. Increased leaf photosynthesis caused by elevated stomatal conductance in a rice mutant deficient in SLAC1, a guard cell anion channel protein.

    Science.gov (United States)

    Kusumi, Kensuke; Hirotsuka, Shoko; Kumamaru, Toshiharu; Iba, Koh

    2012-09-01

    In rice (Oryza sativa L.), leaf photosynthesis is known to be highly correlated with stomatal conductance; however, it remains unclear whether stomatal conductance dominantly limits the photosynthetic rate. SLAC1 is a stomatal anion channel protein controlling stomatal closure in response to environmental [CO(2)]. In order to examine stomatal limitations to photosynthesis, a SLAC1-deficient mutant of rice was isolated and characterized. A TILLING screen of N-methyl-N-nitrosourea-derived mutant lines was conducted for the rice SLAC1 orthologue gene Os04g0674700, and four mutant lines containing mutations within the open reading frame were obtained. A second screen using an infrared thermography camera revealed that one of the mutants, named slac1, had a constitutive low leaf temperature phenotype. Measurement of leaf gas exchange showed that slac1 plants grown in the greenhouse had significantly higher stomatal conductance (g (s)), rates of photosynthesis (A), and ratios of internal [CO(2)] to ambient [CO(2)] (C (i)/C (a)) compared with wild-type plants, whereas there was no significant difference in the response of photosynthesis to internal [CO(2)] (A/C (i) curves). These observations demonstrate that in well-watered conditions, stomatal conductance is a major determinant of photosynthetic rate in rice.

  5. Characterization of a novel high-tillering dwarf 3 mutant in rice

    Institute of Scientific and Technical Information of China (English)

    Bosen Zhang; Feng Tian; Lubin Tan; Daoxin Xie; Chuanqing Sun

    2011-01-01

    Tiller number and culm length are important components of plant architecture and determinate grain production in rice.A line SIL046,derived from an introgression lines population developed by an accession of common wild rice (Oryza rufipogon Griff.) and a high-yielding indica cultivar Guichao 2 (Oryza sativa L.).exhibits a higher tiller number and shorter culm length phenotype than the recipient parent Guichao 2 (GC2).Genetic analysis showed that the high-tillering dwarf phenotype was controlled by a novel single recessive gene,referred to as the high-tillering dwarf3 (htd3),which located within the genetic distance of 13.4 cM between SSR makers RM7003 and RM277 on chromosome 12.By means of fine-mapping strategy,we mapped HTD3 gene within the genetic distance of 2.5 cM and the physical distance of 3100 kb in the centromere of chromosome 12.Further identification of HTD3 gene would provide a new opportunity to uncover the molecular mechanism of the development of culm and tiller,two important components of yields in rice.

  6. 水稻叶色突变体基因定位研究进展%Research Progress on Gene Mapping for Rice Leaf Coloration Mutant

    Institute of Scientific and Technical Information of China (English)

    冯章丽

    2014-01-01

    Rice leaf color mutant is a kind of mutant which is easier to observe and obtain .To fine map the gene controlling the leaf color mutant is benefit to elucidate the regulation mechanism of leaf color gene and genetic improvement of leaf photosynthesis and improve yield of rice .Accordingly ,recent research progress of rice leaf color mutations were reviewed including the types and sources of the leaf coloration mutant ,the genetic mecha-nism of gene mapping and cloning ,molecular mechanism of leaf color mutant and its application in rice was in-troduced ,it was the bases for researching and utilizing the rice leaf color mutation .%水稻叶色突变体是一类较易观察和获得的突变体,研究水稻叶色突变体基因进行定位有助于深入阐明叶色基因的调控机理,对水稻叶片光合遗传改良和产量提高具有重要的指导意义。因此,对于近年来有关水稻叶色突变体的分类来源、遗传机理以及基因定位等研究进展进行综述,并介绍了叶色突变体的分子机理及其在水稻中的应用,旨在为水稻叶色突变体的研究和利用奠定基础。

  7. Loss‐of‐function mutation of rice SLAC7 decreases chloroplast stability and induces a photoprotection mechanism in rice

    OpenAIRE

    Fan, Xiaolei; Wu, Jiemin; Chen, Taiyu; Tie, Weiwei; Chen, Hao; ZHOU, Fei; Lin, Yongjun

    2015-01-01

    Abstract Plants absorb sunlight to power the photochemical reactions of photosynthesis, which can potentially damage the photosynthetic machinery. However, the mechanism that protects chloroplasts from the damage remains unclear. In this work, we demonstrated that rice (Oryza sativa L.) SLAC7 is a generally expressed membrane protein. Loss‐of‐function of SLAC7 caused continuous damage to the chloroplasts of mutant leaves under normal light conditions. Ion leakage indicators related to leaf da...

  8. Role of phenazines and cyclic lipopeptides produced by pseudomonas sp. CMR12a in induced systemic resistance on rice and bean.

    Science.gov (United States)

    Ma, Zongwang; Hua, Gia Khuong Hoang; Ongena, Marc; Höfte, Monica

    2016-08-25

    Pseudomonas sp. CMR12a produces two different classes of cyclic lipopeptides (CLPs) (orfamides and sessilins), which all play a role in direct antagonism against soilborne pathogens. Here we show that Pseudomonas sp. CMR12a is also able to induce systemic resistance to Magnaporthe oryzae on rice and to the web blight pathogen Rhizoctonia solani AG2-2 on bean. Plant assays with biosynthesis mutants of Pseudomonas sp. CMR12a impaired in the production of phenazines and/or CLPs and purified metabolites revealed that distinct bacterial determinants are responsible for inducing systemic resistance in these two pathosystems. In rice, mutants impaired in phenazine production completely lost their ability to induce systemic resistance, while a soil drench with pure phenazine-1-carboxamide (PCN) at a concentration of 0.1 or 1 μM was active in inducing resistance against M. oryzae. In bean, mutants that only produced phenazines, sessilins or orfamides were still able to induce systemic resistance against Rhizoctonia web blight, but a balanced production of these metabolites was needed. This study not only shows that Pseudomonas sp. CMR12a can protect rice to blast disease and bean to web blight disease, but also displays that the determinants involved in induced systemic resistance are plant, pathogen and concentration dependent.

  9. Genetic analysis and gene mapping of a rice few-tillering mutant in early backcross populations (Oryza sativa L.)

    Institute of Scientific and Technical Information of China (English)

    唐家斌; 曾万勇; 王文明; 马炳田; 刘勇; 李浩杰; 夏红爱; 李平; 朱立煌

    2001-01-01

    A rice mutant, G069, characteristic of few tiller numbers, was found in anther culture progeny from the F1 hybrid between an indica-japonica cross, Gui630×02428. The mutant has another two major features: delayed tillering development and yellowing apex and margin on the mature leaves. As a donor parent, G069 was further backcrossed with the recurrent parent, 02428, for two turns to develop a BC2F2 population. Genetic analysis in the BC2F2 population showed that the traits of few-tillering and yellowing apex and margin on the mature leaves were controlled by one recessive gene. A pool of equally mixed genomic DNA, from few-tillering individual plants in BC2F2, was constructed to screen polymorphism with simple sequence repeat (SSR) markers in comparison with the 02428 genome. One SSR marker and three restriction fragment length polymorphism (RFLP) markers were found possibly linked with the recessive gene. By using these markers, the gene of few-tillering was mapped on chromosome 2 between RFLP marker C424 and S13984 with a genetic distance of 2.4 cM and 0.6 cM, respectively. The gene is designated ft1.

  10. Ac/Ds-induced chromosomal rearrangements in rice genomes.

    Science.gov (United States)

    Xuan, Yuan Hu; Zhang, Jianbo; Peterson, Thomas; Han, Chang-Deok

    2012-03-01

    A closely-linked pair of Ac/Ds elements induces chromosomal rearrangements in Arabidopsis and maize. This report summarizes the Ac/Ds systems that generate an exceptionally high frequency of chromosomal rearrangements in rice genomes. From a line containing a single Ds element inserted at the OsRLG5 locus, plants containing a closely-linked pair of inversely-oriented Ds elements were obtained at 1% frequency among the population regenerated from tissue culture. Subsequent regeneration of the lines containing cis-paired Ds elements via tissue culture led to a high frequency (35.6%) of plants containing chromosomal rearrangements at the OsRLG5 locus. Thirty-four rearrangement events were characterized, revealing diverse chromosomal aberrations including deletions, inversions and duplications. Many rearrangements could be explained by sister chromatid transposition (SCT) and homologous recombination (HR), events previously demonstrated in Arabidopsis and maize. In addition, novel events were detected and presumably generated via a new alternative transposition mechanism. This mechanism, termed single chromatid transposition (SLCT), resulted in juxtaposed inversions and deletions on the same chromosome. This study demonstrated that the Ac/Ds system coupled with tissue culture-mediated plant regeneration could induce higher frequencies and a greater diversity of chromosomal rearrangements than previously reported. Understanding transposon-induced chromosomal rearrangements can provide new insights into the relationship between transposable elements and genome evolution, as well as a means to perform chromosomal engineering for crop improvement. Rice is a staple cereal crop worldwide. Complete genome sequencing and rich genetic resources are great advantages for the study of the genomic complexity induced by transposable elements.(1) (-) (2) The combination of tissue culture with genetic lines carrying a pair of closely located Ac/Ds elements greatly increases the

  11. Utilization of induced mutation techniques in rice improvement in the Philippines

    Energy Technology Data Exchange (ETDEWEB)

    Asencion, A.B.; Santos, I.S.; Barrida, A.C.; Medina, F.I.S. III [Philippine Nuclear Research Institute, Atomic Research Center (Philippines)

    2001-03-01

    Rice is one of the most important food crops in the Philippines, as such, efforts have been made consistently to improve the varieties released to the farmers for planting. Both conventional and induced mutation techniques were utilized to solve some of the problems. Varieties with improved qualities, resistance to pests and diseases, reduced height, early maturity and non-photoperiod sensitive were developed using either physical or chemical mutagen. Other methods were also tried to enhance variability like combination of gamma irradiation and biotechnology. Irradiation of F1 seeds was also initiated including the use of mutants in crosses to transfer their improved mutated characters. Promising selections were already tested and six lines from the F1 irradiation out-yielded both C4-63G and IR 1561-288-3, their parents. (author)

  12. Overexpression of Arabidopsis and rice stress genes' inducible transcription factor confers drought and salinity tolerance to rice.

    Science.gov (United States)

    Datta, Karabi; Baisakh, Niranjan; Ganguly, Moumita; Krishnan, Sellapan; Yamaguchi Shinozaki, Kazuko; Datta, Swapan K

    2012-06-01

    Rice yield is greatly affected by environmental stresses such as drought and salinity. In response to the challenge of producing rice plants tolerant to these stresses, we introduced cDNA encoding the transcription factors DREB1A and DREB1B under the control of the stress inducible rd29 promoter. Two different indica rice cultivars were used, BR29, an improved commercially cultivated variety from Bangladesh and IR68899B, an IRRI bred maintainer line for hybrid rice. Agrobacterium mediated transformation of BR29 was done independently with DREB1A isolated from rice and Arabidopsis and DREB1B isolated from rice, whereas biolistic transformation was done with rice- DREB1B in the case of IR68899B. Initial genetic integration was confirmed by PCR and Southern blot analysis. Salinity tolerance was assayed in very young seedlings. Drought stress tests were found to be more reliable when they were carried out at the pre-flowering booting stage. RNA gel blot analysis as well as quantitative PCR analysis was performed to estimate the transcription level under stressed and unstressed conditions. Agronomic performance studies were done with stressed and unstressed plants to compare the yield losses due to dehydration and salt loading stresses. Noticeably enhanced tolerance to dehydration was observed in the plants transformed with DREB1A isolated from Arabidopsis while DREB1B was found to be more effective for salt tolerance.

  13. Plant Hormone Resistance and Agronomic Characteristics of the MT10 Mutant in Rice

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    The MT10 mutant plants had resistances to auxin. Under light and dark culture,the roots of MT10 seedlings had shown less lateral roots and short lateral roots. In soil ,MT10 seedlings had shown not only no changed agronomic characteristics but also no significant difference with WT.

  14. Inducement and identification of an endosperm mutant in maize

    African Journals Online (AJOL)

    ajl yemi

    2011-11-30

    Nov 30, 2011 ... “super sweet” phenotype were derived from the mutated offspring. ... characteristics and distinguished molecular mechanism to the previous mutants of gene sh2, these three mutant lines are ...... Physical association of starch biosynthetic ... reduced seedling mutant in oilseed rape, Brassica napus, for.

  15. Cell wall degrading enzyme induced rice innate immune responses are suppressed by the type 3 secretion system effectors XopN, XopQ, XopX and XopZ of Xanthomonas oryzae pv. oryzae.

    Science.gov (United States)

    Sinha, Dipanwita; Gupta, Mahesh Kumar; Patel, Hitendra Kumar; Ranjan, Ashish; Sonti, Ramesh V

    2013-01-01

    Innate immune responses are induced in plants and animals through perception of Damage Associated Molecular Patterns. These immune responses are suppressed by pathogens during infection. A number of studies have focussed on identifying functions of plant pathogenic bacteria that are involved in suppression of Pathogen Associated Molecular Pattern induced immune responses. In comparison, there is very little information on functions used by plant pathogens to suppress Damage Associated Molecular Pattern induced immune responses. Xanthomonasoryzae pv. oryzae, a gram negative bacterial pathogen of rice, secretes hydrolytic enzymes such as LipA (Lipase/Esterase) that damage rice cell walls and induce innate immune responses. Here, we show that Agrobacterium mediated transient transfer of the gene for XopN, a X. oryzae pv. oryzae type 3 secretion (T3S) system effector, results in suppression of rice innate immune responses induced by LipA. A xopN (-) mutant of X. oryzae pv. oryzae retains the ability to suppress these innate immune responses indicating the presence of other functionally redundant proteins. In transient transfer assays, we have assessed the ability of 15 other X. oryzae pv. oryzae T3S secreted effectors to suppress rice innate immune responses. Amongst these proteins, XopQ, XopX and XopZ are suppressors of LipA induced innate immune responses. A mutation in any one of the xopN, xopQ, xopX or xopZ genes causes partial virulence deficiency while a xopN (-) xopX (-) double mutant exhibits a greater virulence deficiency. A xopN (-) xopQ (-) xopX (-) xopZ (-) quadruple mutant of X. oryzae pv. oryzae induces callose deposition, an innate immune response, similar to a X. oryzae pv. oryzae T3S(-) mutant in rice leaves. Overall, these results indicate that multiple T3S secreted proteins of X. oryzae pv. oryzae can suppress cell wall damage induced rice innate immune responses.

  16. Identification of a phosphinothricin-resistant mutant of rice glutamine synthetase using DNA shuffling.

    Science.gov (United States)

    Tian, Yong-Sheng; Xu, Jing; Zhao, Wei; Xing, Xiao-Juan; Fu, Xiao-Yan; Peng, Ri-He; Yao, Quan-Hong

    2015-10-23

    To date, only bar/pat gene derived from Streptomyces has been used to generate the commercial PPT-resistant crops currently available in the market. The limited source of bar/pat gene is probably what has caused the decrease in PPT-tolerance, which has become the main concern of those involved in field management programs. Although glutamine synthetase (GS) is the target enzyme of PPT, little study has been reported about engineering PPT-resistant plants with GS gene. Then, the plant-optimized GS gene from Oryza sativa (OsGS1S) was chemically synthesized in the present study by PTDS to identify a GS gene for developing PPT-tolerant plants. However, OsGS1S cannot be directly used for developing PPT-tolerant plants because of its poor PPT-resistance. Thus, we performed DNA shuffling on OsGS1S, and one highly PPT-resistant mutant with mutations in four amino acids (A63E, V193A, T293A and R295K) was isolated after three rounds of DNA shuffling and screening. Among the four amino acids substitutions, only R295K was identified as essential in altering PPT resistance. The R295K mutation has also never been previously reported as an important residue for PPT resistance. Furthermore, the mutant gene has been transformed into Saccharomyces cerevisiae and Arabidopsis to confirm its potential in developing PPT-resistant crops.

  17. LC2 and OsVIL2 Promote Rice Flowering by Photoperoid-Induced Epigenetic Silencing of OsLF

    Institute of Scientific and Technical Information of China (English)

    Jun Wang; Jiang Hu; Qian Qian; Hong-Wei Xue

    2013-01-01

    Proper flowering time is essential for plant reproduction.Winter annual Arabidopsis thaliana needs vernalization before flowering,during which AtVlLs (VlN3 and VRN5,components of PRC2 complex) mediate the H3K27 trimethylation at the FLC locus (a floral repressor) to repress the FLC expression and hence to induce flowering.However,how VlLs (VlL,VERNALIZATION INSENSITIVE 3-LIKE) function in rice is unknown.Here we demonstrated that rice LC2 (OsVlL3) and OsVll.2 (two OsVlLs,possible components of PRC2 complex) promote rice flowering.Our results showed that expressions of LC2 and OsVlL2 are induced by SD (short-day) conditions and both Ic2 mutant and OsVlL2-RNAi lines display delayed heading date,consistent with the reduced expression levels of Hdl and Hd3a.Interestingly,LC2 binds to the promoter region of a floral repressor OsLF and represses the OsLF expression via H3K27 tri-methylation modification.In addition,OsLF directly regulates the Hdl expression through binding to Hdl promoter.These results first demonstrated that the putative PRC2 in rice is involved in photoperiod flowering regulation,which is different from that of Arabidopsis,and revealed that LC2 binds the promoter region of target gene,presenting a possible mechanism of the recruitment process of PRC2 complex to its target genes.The studies provide informative clues on the epigenetic control of rice flowering.

  18. Morphological and starch structural characteristics of the Japonica rice mutant variety Seolgaeng for dry-milled flour

    Science.gov (United States)

    Producing fine, good quality rice flour is more difficult than wheat flour because the rice grain is harder. In this study, we analyzed the relationship between the morphology and starch of kernels from genetically different rice varieties that can be used to make dry-milled flour. The non-glutinous...

  19. Consumption of Mercury-contaminated Rice Induces Oxidative Stress and Free Radical Aggravation in Rats

    Institute of Scientific and Technical Information of China (English)

    XIU-LING JI; GUI-WEN JIN; JIN-PING CHENG; WEN-HUA WANG; JING LU; LI-YA QU

    2007-01-01

    Objective To study the oxidative stress induced by consumption of mercury-contaminated rice in rats, and to assess the possible public health risk of mercury contamination in Wanshan mining area. Methods Sprague Dawley rats were fed the mercury-contaminated rice produced from Wanshan area for 90 days. The antioxidant status and the free radicals in rat serum were evaluated. Results High mercury accumulation in organs of rats fed the mercury-contaminated rice confirmed the server pollution of mercury in Wanshan mining area. The intensity of electron spin resonance (ESR) signal increased by 87.38% in rats fed the rice from Wanshan compared with that in the control rats fed the rice from Shanghai, suggesting that chronic dietary consumption of rice from mercury mining area could induce an aggravation of free radicals. Feeding the mercury-contaminated rice was associated with significant decreases in the antioxidant enzymatic activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) and concentration of serum nitric oxide (NO), but it had no effect on serum nitric oxide synthase (NOS) activity. Feeding the mercury-contaminated rice raised the level of serum malonyldialdehyde (MDA), indicating the occurrence of oxidative stress. Conclusion The long-term dietary consumption of mercury-contaminated rice induces the aggravation of free radicals and exerts oxidative stress.

  20. Generation of transgenic rice lines with reduced contents of multiple potential allergens using a null mutant in combination with an RNA silencing method.

    Science.gov (United States)

    Wakasa, Yuhya; Hirano, Kana; Urisu, Atsuo; Matsuda, Tsukasa; Takaiwa, Fumio

    2011-12-01

    Rice seed proteins are known to be a causative antigen in some patients with food allergy, especially cereal allergy, with clinical symptoms such as eczema and dermatitis. The α-amylase/trypsin inhibitors (14-16 kDa), α-globulin (26 kDa) and β-glyoxalase I (33 kDa) are regarded as major potential allergens of rice (Oryza sativa L.) seed based on specific recognition by serum IgE from allergy patients. In order to suppress the production of these major allergens in rice grains, a mutant in the 'Koshihikari' background lacking the 26 kDa allergen (GbN-1) was used as a host for RNA silencing. A binary vector harboring two RNA interference (RNAi) gene cassettes for suppression of 14-16 kDa and 33 kDa allergens driven by the 13 kDa and 10 kDa prolamin endosperm-specific promoters, respectively, was introduced into the GbN-1 genome by Agrobacterium-mediated transformation. In the most promising transgenic line, the content of the three potential allergens was remarkably reduced to a very faint level without a change in seed phenotype. IgE binding of 15 patients' sera to the transgenic rice seed mostly deficient in the three major allergens was on average only about 10% that of the control wild-type rice, suggesting that these three accounted for the great majority of rice seed causative allergens recognized by patients' IgE and that the sequential allergen deletion/reduction strategy works in the development of hypo-allergenic rice lines.

  1. 2个水稻矮杆突变体的遗传分析%Genetic Analysis of Two Dwarf Mutants of Rice

    Institute of Scientific and Technical Information of China (English)

    薛建峰; 汤永荣; 查仁明; 罗洪发

    2011-01-01

    The aim was to study the genetic development of dwarf mutants of rice. Two dwarf mutants mini-plant (mp) and dwarf multi-tiller, reciprocal crossed with 'Minghui 63' and 'Minghui 86'. By the character investigation and chi-square test, in the F1 generation, two dwarf mutants had recessive heredity. In the F2 segregating population, the number of rice dwarf mutant and wild-type plants, met Mendel's single-genetic laws. Therefore, it confirmed that the two dwarf mutants of rice were controlled by a single autosomal recessive gene, respectively.%为了研究水稻矮杆突变体的遗传规律,通过将2个矮杆突变体mini-plant(mp)和矮蘖丛生突变体分别与‘明恢63’、‘明恢86’等进行正反交杂交组合,经过田间的性状调查与卡方测定,发现F1代中2个矮杆突变体均为隐性遗传,在F2分离群体中,2个矮杆突变体的遗传规律符合孟德尔的常染色体单基因隐性遗传.因此,可以推断2个水稻矮杆突变体都是由常染色体上的隐性单基因控制.

  2. The phenome analysis of mutant alleles in Leucine-Rich Repeat Receptor-Like Kinase genes in rice reveals new potential targets for stress tolerant cereals.

    Science.gov (United States)

    Dievart, Anne; Perin, Christophe; Hirsch, Judith; Bettembourg, Mathilde; Lanau, Nadège; Artus, Florence; Bureau, Charlotte; Noel, Nicolas; Droc, Gaétan; Peyramard, Matthieu; Pereira, Serge; Courtois, Brigitte; Morel, Jean-Benoit; Guiderdoni, Emmanuel

    2016-01-01

    Plants are constantly exposed to a variety of biotic and abiotic stresses that reduce their fitness and performance. At the molecular level, the perception of extracellular stimuli and the subsequent activation of defense responses require a complex interplay of signaling cascades, in which protein phosphorylation plays a central role. Several studies have shown that some members of the Leucine-Rich Repeat Receptor-Like Kinase (LRR-RLK) family are involved in stress and developmental pathways. We report here a systematic analysis of the role of the members of this gene family by mutant phenotyping in the monocotyledon model plant rice, Oryza sativa. We have then targeted 176 of the ∼320 LRR-RLK genes (55.7%) and genotyped 288 mutant lines. Position of the insertion was confirmed in 128 lines corresponding to 100 LRR-RLK genes (31.6% of the entire family). All mutant lines harboring homozygous insertions have been screened for phenotypes under normal conditions and under various abiotic stresses. Mutant plants have been observed at several stages of growth, from seedlings in Petri dishes to flowering and grain filling under greenhouse conditions. Our results show that 37 of the LRR-RLK rice genes are potential targets for improvement especially in the generation of abiotic stress tolerant cereals.

  3. Genetic analysis and gene mapping of leafy head (lhd), a mutant blocking the differentiation of rachis branches in rice (Oryza sativa L.)

    Institute of Scientific and Technical Information of China (English)

    DUAN Yuanlin; MAO Damei; LI Weiming; XUE Yongbiao; WU Weiren; LIU Huaqing; ZHANG Danfeng; ZHOU Yuanchang; PAN Runsheng; LIN Lihui; CHEN Zhiwei; GUAN Huazhong

    2003-01-01

    A rice mutant called leafy head (lhd), in which the differentiation of rachis branches is blocked, was identified in a doubled haploid (DH) population derived through F1 anther culture from a cross between rice (Oryza sativa L.) indica cultivar Gui-630 and japonica cultivar Taiwanjing. The mutant is shorter in plant height, possessing smaller and clumpy leaves, and always stays at the vegetative growth stage. Genetic analysis suggests that lhd is controlled by a single recessive gene, which is temporarily named lhd(t). The phenotype of the mutant suggests that LHD(t) is a key gene controlling the differentiation of rachis branches. In order to map the gene, two F2 populations were constructed by crossing the lhd heterozygote with varieties Minghui-77 (indica) and Jinghua-8 (japonica). In the F2 of lhd heterozygote ( Jinghua-8, some mutant plants appeared as the "medium type", suggesting that the lhd phenotype could be influenced by genetic backgrounds. With the published SSR markers of RM series and additional SSR markers developed by ourselves and using the methods of bulked segregant analysis (BSA) and mutant analysis (with 498 mutant plants in total), LHD(t) gene was mapped onto the distal region of the long arm of chromosome 10. Markers SSR1, RM269, RM258, RM304 and RM171 were located on one side with distances of 6.4, 16.6, 18.4, 22.2 and 26.3 cM to LHD(t); whereas markers SSR4 and SSR5 were on the other side with distances of 0.6 and 2.2 cM to LHD(t). The results will facilitate the positional cloning and functional study of the LHD(t) gene.

  4. Selection of mutants of capsicum annuum induced by gamma ray

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Y. I.; Lee, Y. B. [Korea Atomic Energy Research Institute, Taejeon (Korea, Republic of); Lee, E. K. [Chungnam National Univ., Taejeon (Korea, Republic of)

    1998-06-01

    For induction and selection of mutations of Capsicum annuum L., dry seeds of pure lines No.1 and No.2 were irradiated with gamma ray of 150Gy, 200Gy and 250Gy. Various mutants were selected such as showing early maturity, short plant height, long fruit and chlorophyll mutations. Mutation frequency of No.1 line was 3.4% in the dose of 150Gy, while the frequency of No.2 line was 2.7% in the dose of 250Gy. For selection of resistant mutant to amino acid analog, the optimum concentration of 5-methyltryptophan (5-MT) and S-(2-aminoethyl)-L-cysteine were 25 ppm and 30 ppm, respectively. Four resistant mutant lines to 5-MT were selected among 400 mutant lines.

  5. Hepatoprotective effects of rice-derived peptides against acetaminophen-induced damage in mice.

    Science.gov (United States)

    Kawakami, Kayoko; Moritani, Chie; Uraji, Misugi; Fujita, Akiko; Kawakami, Koji; Hatanaka, Tadashi; Suzaki, Etsuko; Tsuboi, Seiji

    2017-03-01

    Glutathione, the most abundant intracellular antioxidant, protects cells against reactive oxygen species induced oxidative stress and regulates intracellular redox status. We found that rice peptides increased intracellular glutathione levels in human hepatoblastoma HepG2 cells. Acetaminophen is a commonly used analgesic. However, an overdose of acetaminophen causes severe hepatotoxicity via depletion of hepatic glutathione. Here, we investigated the protective effects of rice peptides on acetaminophen-induced hepatotoxicity in mice. ICR mice were orally administered rice peptides (0, 100 or 500 mg/kg) for seven days, followed by the induction of hepatotoxicity via intraperitoneal injection of acetaminophen (700 mg/kg). Pretreatment with rice peptides significantly prevented increases in serum alanine aminotransferase, aspartate aminotransferase, and lactate dehydrogenase levels and protected against hepatic glutathione depletion. The expression of γ-glutamylcysteine synthetase, a key regulatory enzyme in the synthesis of glutathione, was decreased by treatment with acetaminophen, albeit rice peptides treatment recovered its expression compared to that achieved treatment with acetaminophen. In addition, histopathological evaluation of the livers also revealed that rice peptides prevented acetaminophen-induced centrilobular necrosis. These results suggest that rice peptides increased intracellular glutathione levels and could protect against acetaminophen-induced hepatotoxicity in mice.

  6. Production of superoxide from photosystem II-light harvesting complex II supercomplex in STN8 kinase knock-out rice mutants under photoinhibitory illumination.

    Science.gov (United States)

    Poudyal, Roshan Sharma; Nath, Krishna; Zulfugarov, Ismayil S; Lee, Choon-Hwan

    2016-09-01

    When phosphorylation of Photosystem (PS) II core proteins is blocked in STN8 knock-out mutants of rice (Oryza sativa) under photoinhibitory illumination, the mobilization of PSII supercomplex is prevented. We have previously proposed that more superoxide (O2(-)) is produced from PSII in the mutant (Nath et al., 2013, Plant J. 76, 675-686). Here, we clarify the type and site for the generation of reactive oxygen species (ROS). Using both histochemical and fluorescence probes, we observed that, compared with wild-type (WT) leaves, levels of ROS, including O2(-) and hydrogen peroxide (H2O2), were increased when leaves from mutant plants were illuminated with excess light. However, singlet oxygen production was not enhanced under such conditions. When superoxide dismutase was inhibited, O2(-) production was increased, indicating that it is the initial event prior to H2O2 production. In thylakoids isolated from WT leaves, kinase was active in the presence of ATP, and spectrophotometric analysis of nitrobluetetrazolium absorbance for O2(-) confirmed that PSII-driven superoxide production was greater in the mutant thylakoids than in the WT. This contrast in levels of PSII-driven superoxide production between the mutants and the WT plants was confirmed by conducting protein oxidation assays of PSII particles from osstn8 leaves under strong illumination. Those assays also demonstrated that PSII-LHCII supercomplex proteins were oxidized more in the mutant, thereby implying that PSII particles incur greater damage even though D1 degradation during PSII-supercomplex mobilization is partially blocked in the mutant. These results suggest that O2(-) is the major form of ROS produced in the mutant, and that the damaged PSII in the supercomplex is the primary source of O2(-).

  7. Structural characterization of a mixed-linkage glucan deficient mutant reveals alteration in cellulose microfibril orientation in rice coleoptile mesophyll cell walls

    Directory of Open Access Journals (Sweden)

    Andreia Michelle Smith-Moritz

    2015-08-01

    Full Text Available The CELLULOSE SYNTHASE-LIKE F6 (CslF6 gene was previously shown to mediate the biosynthesis of mixed-linkage glucan (MLG, a cell wall polysaccharide that is hypothesized to be a tightly associated with cellulose and also have a role in cell expansion in the primary cell wall of young seedlings in grass species. We have recently shown that loss-of-function cslf6 rice mutants do not accumulate MLG in most vegetative tissues. Despite the absence of a structurally important polymer, MLG, these mutants are unexpectedly viable and only show a moderate growth compromise compared to wild type. Therefore these mutants are ideal biological systems to test the current grass cell wall model. In order to gain a better understanding of the role of MLG in the primary wall, we performed in-depth compositional and structural analyses of the cell walls of three day-old rice seedlings using various biochemical and novel microspectroscopic approaches. We found that cellulose content as well as matrix polysaccharide composition was not significantly altered in the MLG deficient mutant. However, we observed a significant change in cellulose microfibril bundle organization in mesophyll cell walls of the cslf6 mutant. Using synchrotron source Fourier Transform Mid-Infrared Spectromicroscopy for high-resolution imaging, we determined that the bonds associated with cellulose and arabinoxylan, another major component of the primary cell was of grasses, were in a lower energy configuration compared to wild type, suggesting a slightly weaker primary wall in MLG deficient mesophyll cells. Taken together, these results suggest that MLG may influence cellulose deposition in mesophyll cell walls without significantly affecting anisotropic growth thus challenging MLG importance in cell wall expansion.

  8. Diclofop-methyl affects microbial rhizosphere community and induces systemic acquired resistance in rice.

    Science.gov (United States)

    Chen, Si; Li, Xingxing; Lavoie, Michel; Jin, Yujian; Xu, Jiahui; Fu, Zhengwei; Qian, Haifeng

    2017-01-01

    Diclofop-methyl (DM), a widely used herbicide in food crops, may partly contaminate the soil surface of natural ecosystems in agricultural area and exert toxic effects at low dose to nontarget plants. Even though rhizosphere microorganisms strongly interact with root cells, little is known regarding their potential modulating effect on herbicide toxicity in plants. Here we exposed rice seedlings (Xiushui 63) to 100μg/L DM for 2 to 8days and studied the effects of DM on rice rhizosphere microorganisms, rice systemic acquired resistance (SAR) and rice-microorganisms interactions. The results of metagenomic 16S rDNA Illumina tags show that DM increases bacterial biomass and affects their community structure in the rice rhizosphere. After DM treatment, the relative abundance of the bacterium genera Massilia and Anderseniella increased the most relative to the control. In parallel, malate and oxalate exudation by rice roots increased, potentially acting as a carbon source for several rhizosphere bacteria. Transcriptomic analyses suggest that DM induced SAR in rice seedlings through the salicylic acid (but not the jasmonic acid) signal pathway. This response to DM stress conferred resistance to infection by a pathogenic bacterium, but was not influenced by the presence of bacteria in the rhizosphere since SAR transcripts did not change significantly in xenic and axenic plant roots exposed to DM. The present study provides new insights on the response of rice and its associated microorganisms to DM stress. Copyright © 2016. Published by Elsevier B.V.

  9. Alpha-picolinic acid,a fungal toxin and mammal apoptosis-inducing agent,elicits hypersensitive-like response and enhances disease resistance in rice

    Institute of Scientific and Technical Information of China (English)

    Hai Kuo ZHANG; Xin ZHANG; Bi Zeng MAO; Qun LI; Zu Hua HE

    2004-01-01

    Alpha-picolinic acid (PA),a metabolite of tryptophan and an inducer of apoptosis in the animal cell,has been reported to be a toxin produced by some of plant fungal pathogens and used in screening for disease resistant mutants. Here,we report that PA is an efficient apoptosis agent triggering cell death of hypersensitive-like response in planta. Confirmed by Fluorescence Activated Cell Sorter (FACS),rice suspension cells and leaves exhibited programmed cell death induced by PA. The PA-induced cell death was associated with the accumulation of reactive oxygen species that could be blocked by diphenylene iodonium chloride,indicating that the generation of reactive oxygen species was NADPHoxidase dependent. We also demonstrated the induction of rice defense-related genes and subsequent resistant enhancement by PA against the rice blast fungus Magnaporthe grisea. Hence,it was concluded that the PA-stimulated defense response likely involves the onset of the hypersensitive response in rice,which also provides a simple eliciting tool for studying apoptosis in the plant cell.

  10. Decreased photosynthesis in the erect panicle 3 (ep3) mutant of rice is associated with reduced stomatal conductance and attenuated guard cell development

    OpenAIRE

    2015-01-01

    The ERECT PANICLE 3 gene of rice encodes a peptide that exhibits more than 50% sequence identity with the Arabidopsis F-box protein HAWAIIAN SKIRT (HWS). Ectopic expression of the Os02g15950 coding sequence, driven by the HWS (At3g61950) promoter, rescued the hws-1 flower phenotype in Arabidopsis confirming that EP3 is a functional orthologue of HWS. In addition to displaying an erect inflorescence phenotype, loss-of-function mutants of Os02g15950 exhibited a decrease in leaf photosynthetic c...

  11. Genetic analysis and gene mapping of a rice few-tillering mutant in early backcross populations (Oryza sativa L.)

    Institute of Scientific and Technical Information of China (English)

    TANG; Jiabin; (

    2001-01-01

    [1]Xu, Y. B., Shen, Z. T., Diallel analysis of tiller number at different growth stages in rice (Oryza sativa L.), Theoretical and Applied Genetics, 1991, 83(2): 243-249.[2]Wu, W. R., Li, W. M., Tang, D. Z. et al., Time-related mapping of quantitative trait loci underlying tiller number in rice, Genetics, 1999, (151): 297-303.[3]Wu, P., Zhang, G ., Huang, N., Identification of QTLs controlling quantitative characters in rice using RFLP markers, Euphytica, 1996, 89(3): 349-354.[4]Khush, G . S., Breaking the yield frontier of rice, GeoJournal, 1995, 35(3): 329-332.[5]Chen, X., Themnykh, S., Xu, Y. et al., Development of microsatellite framework providing genome-wide coverage in rice (Oryzae sativa L.), Theoretical and Applied Genetics, 1997, 95(4): 553-567.[6]Panaud, O., Chen, X., McCouch, S. R. et al., Development of microsatellite markers and characterization of simple sequence length polymorphisms (SSLPs) in rice (Oryzae sativa L.), Molecular and General Genetics, 1996, (252): 597-607.[7]McCouth, S. R., Kochert, G., Yu, Z. H. et al., Molecular mapping of rice chromosomes, Theoretical and Applied Genetics, 1988, 76: 815-829.[8]Harushima, Y., Yano, M., Shomura, A. et al., A high-density rice genetic linkage map with 2275 markers using a single F2 population, Genetics, 1998, 148: 479-494.[9]Lander, E. S. et al., Mapmaker: An interactive computer for construction primary genetics linkage maps of experimental and natural populations, Genomics, 1987, (1): 174-182.[10]Kinoshita, T., Report of committee on gene symbolization, nomenclature and linkage groups, Rice Genetic Newsletter, 1995, (12): 9-153.

  12. Relationship between gene responses and symptoms induced by Rice grassy stunt virus

    Directory of Open Access Journals (Sweden)

    Kouji eSatoh

    2013-10-01

    Full Text Available Rice grassy stunt virus (RGSV is a serious threat to rice production in Southeast Asia. RGSV is a member of the genus Tenuivirus, and it induces leaf yellowing, stunting, and excess tillering on rice plants. Here we examined gene responses of rice to RGSV infection to gain insight into the gene responses which might be associated with the disease symptoms. The results indicated that 1 many genes related to cell wall synthesis and chlorophyll synthesis were predominantly suppressed by RGSV infection; 2 RGSV infection induced genes associated with tillering process; 3 RGSV activated genes involved in inactivation of gibberellic acid and indole-3-acetic acid ; and 4 the genes for strigolactone signaling were suppressed by RGSV. These results suggest that these gene responses to RGSV infection account for the excess tillering specific to RGSV infection as well as other symptoms by RGSV, such as stunting and leaf chlorosis.

  13. Calreticulin mutants in mice induce an MPL-dependent thrombocytosis with frequent progression to myelofibrosis.

    Science.gov (United States)

    Marty, Caroline; Pecquet, Christian; Nivarthi, Harini; El-Khoury, Mira; Chachoua, Ilyas; Tulliez, Micheline; Villeval, Jean-Luc; Raslova, Hana; Kralovics, Robert; Constantinescu, Stefan N; Plo, Isabelle; Vainchenker, William

    2016-03-10

    Frameshift mutations in the calreticulin (CALR) gene are seen in about 30% of essential thrombocythemia and myelofibrosis patients. To address the contribution of the CALR mutants to the pathogenesis of myeloproliferative neoplasms, we engrafted lethally irradiated recipient mice with bone marrow cells transduced with retroviruses expressing these mutants. In contrast to wild-type CALR, CALRdel52 (type I) and, to a lesser extent, CALRins5 (type II) induced thrombocytosis due to a megakaryocyte (MK) hyperplasia. Disease was transplantable into secondary recipients. After 6 months, CALRdel52-, in contrast to rare CALRins5-, transduced mice developed a myelofibrosis associated with a splenomegaly and a marked osteosclerosis. Monitoring of virus-transduced populations indicated that CALRdel52 leads to expansion at earlier stages of hematopoiesis than CALRins5. However, both mutants still specifically amplified the MK lineage and platelet production. Moreover, a mutant deleted of the entire exon 9 (CALRdelex9) did not induce a disease, suggesting that the oncogenic property of CALR mutants was related to the new C-terminus peptide. To understand how the CALR mutants target the MK lineage, we used a cell-line model and demonstrated that the CALR mutants, but not CALRdelex9, specifically activate the thrombopoietin (TPO) receptor (MPL) to induce constitutive activation of Janus kinase 2 and signal transducer and activator of transcription 5/3/1. We confirmed in c-mpl- and tpo-deficient mice that expression of Mpl, but not of Tpo, was essential for the CALR mutants to induce thrombocytosis in vivo, although Tpo contributes to disease penetrance. Thus, CALR mutants are sufficient to induce thrombocytosis through MPL activation. © 2016 by The American Society of Hematology.

  14. Mutant connexin 50 (S276F) inhibits channel and hemichannel functions inducing cataract

    Indian Academy of Sciences (India)

    Yuanyuan Liu; Chen Qiao; Tanwei Wei; Fang Zheng; Shuren Guo; Qiang Chen; Ming Yan; Xin Zhou

    2015-06-01

    This study was designed to detect the expression, detergent resistance, subcellular localization, and channel and hemichannel functions of mutant Cx50 to understand the forming mechanism for inducing congenital cataract by a novel mutation p.S276F in connexin 50 (Cx50) reported previously by us. HeLa and human lens epithelial (HLE) cells were transfected with wild-type Cx50 and mutant Cx50 (S276F). We examined the functional characteristics of mutant Cx50 (S276F) in comparison with those of wild-type Cx50 using immunoblot, confocal fluorescence microscopy, dye transfer analysis and dye uptake assay. The mutant and wild-type Cx50 were expressed in equal levels and could efficiently localize to the plasma membrane without transportation and assembly problems. Scrape loading dye transfer was significantly evident in cells transfected with wild-type Cx50 compared to those in cells transfected with mutant Cx50 and cotransfected with wild-type and mutant Cx50. The dye uptake was found to be significantly lower in cells transfected with mutant Cx50 than in cells transfected with wild-type Cx50 and cells cotransfected with wild-type and mutant Cx50. The transfected HeLa and HLE cell lines showed similar performance in all the experiments. These results indicated that the mutant Cx50 (S276F) might inhibit the function of gap junction channel in a dominant negative manner, but inhibit the hemichannel function in a recessive negative manner.

  15. Down-regulation of OsSAG12-1 results in enhanced senescence and pathogen-induced cell death in transgenic rice plants

    Indian Academy of Sciences (India)

    Subaran Singh; Mrunmay Kumar Giri; Praveen Kumar Singh; Adnan Siddiqui; Ashis Kumar Nandi

    2013-09-01

    Senescence is a highly regulated process accompanied by changes in gene expression. While the mRNA levels of most genes decline, the mRNA levels of specific genes (senescence associated genes, SAGs) increase during senescence. Arabidopsis SAG12 (AtSAG12) gene codes for papain-like cysteine protease. The promoter of AtSAG12 is SA-responsive and reported to be useful to delay senescence by expressing cytokinin biosynthesis gene isopentenyltransferase specifically during senescence in several plants including Arabidopsis, lettuce and rice. The physiological role of AtSAG12 is not known; the homozygous atsag12 mutant neither fails to develop senescence-associated vacuoles nor shows any morphological phenotype. Through BLAST search using AtSAG12 amino acid sequences as query, we identified a few putative homologues from rice genome (OsSAGs; Oryza sativa SAGs). OsSAG12-1 is the closest homologue of AtSAG12 with 64% similar amino acid composition. Expression of OsSAG12-1 is induced during senescence and pathogen-induced cell death. To evaluate the possible role of OsSAG12-1 we generated RNAi transgenic lines in Japonica rice cultivar TP309. The transgenic lines developed early senescence at varying levels and showed enhanced cell death when inoculated with bacterial pathogen Xanthomonas oryzae pv.oryzae. Our results suggest that OsSAG12-1 is a negative regulator of cell death in rice.

  16. Rice improvement through radiation-induced mutation for cultivation in South Vietnam

    Energy Technology Data Exchange (ETDEWEB)

    Do Khac Thinh; Hung Phi Oanh; Nguyen Thi Cuc; Nguyen Ngoc Quynh [Institute of Agricultural Science of South Vietnam, Ho Chi Minh (Viet Nam)

    2001-03-01

    For past years, rice varieties cultivated in South Vietnam originated from domestic hybridisation or from IRRI. Rice mutation breeding has been initiated for recent years. To meet the requirement of rice production diversification in different agro-ecological areas and rice genetic resources, from 1993 Institute of Agricultural Science of South Vietnam has carried out rice improvement by induced mutation of radiation. The mutagen was gamma rays of {sup 60}Co. The goal is to create inherited variations, which cannot be obtained from other breeding methods, specially important characters of rice varieties (high tolerance to acid sulfate soil, lodging resistance combined with early maturity), which were difficult to gain by hybridisation. With {sup 60}Co gamma rays, doses of 10-20 krad, dose rate of 280 krad/h, dry and germinated seeds of introduced and local rice varieties (IR 64, IR 9729, IR 50404, IR 59606, Jasmine 85, Nang Huong, Tam Xoan) were irradiated. The irradiated seeds were immediately sown within 24 and 94 hrs for wet seeds and dry seeds after treatment, respectively. Population of 10,000-15,000 plants were established and evaluated by IRRI evaluation standard from M2-M7 generations. 365 lines, varieties were selected with better behaviours than original varieties as lodging resistance, earliness, potential yield, leaf characters, tolerant ability to adverse conditions etc. Some good varieties (VND95-19, VND95-20) have been approved as leading national varieties and released for large-scale production in South Vietnam. (author)

  17. Reverse genetic screening reveals poor correlation between morpholino-induced and mutant phenotypes in zebrafish.

    Science.gov (United States)

    Kok, Fatma O; Shin, Masahiro; Ni, Chih-Wen; Gupta, Ankit; Grosse, Ann S; van Impel, Andreas; Kirchmaier, Bettina C; Peterson-Maduro, Josi; Kourkoulis, George; Male, Ira; DeSantis, Dana F; Sheppard-Tindell, Sarah; Ebarasi, Lwaki; Betsholtz, Christer; Schulte-Merker, Stefan; Wolfe, Scot A; Lawson, Nathan D

    2015-01-12

    The widespread availability of programmable site-specific nucleases now enables targeted gene disruption in the zebrafish. In this study, we applied site-specific nucleases to generate zebrafish lines bearing individual mutations in more than 20 genes. We found that mutations in only a small proportion of genes caused defects in embryogenesis. Moreover, mutants for ten different genes failed to recapitulate published Morpholino-induced phenotypes (morphants). The absence of phenotypes in mutant embryos was not likely due to maternal effects or failure to eliminate gene function. Consistently, a comparison of published morphant defects with the Sanger Zebrafish Mutation Project revealed that approximately 80% of morphant phenotypes were not observed in mutant embryos, similar to our mutant collection. Based on these results, we suggest that mutant phenotypes become the standard metric to define gene function in zebrafish, after which Morpholinos that recapitulate respective phenotypes could be reliably applied for ancillary analyses.

  18. Treatment of GABA from Fermented Rice Germ Ameliorates Caffeine-Induced Sleep Disturbance in Mice.

    Science.gov (United States)

    Mabunga, Darine Froy N; Gonzales, Edson Luck T; Kim, Hee Jin; Choung, Se Young

    2015-05-01

    γ-Aminobutyric acid (GABA), a major inhibitory neurotransmitter in the mammalian central nervous system, is involved in sleep physiology. Caffeine is widely used psychoactive substance known to induce wakefulness and insomnia to its consumers. This study was performed to examine whether GABA extracts from fermented rice germ ameliorates caffeine-induced sleep disturbance in mice, without affecting spontaneous locomotor activity and motor coordination. Indeed, caffeine (10 mg/kg, i.p.) delayed sleep onset and reduced sleep duration of mice. Conversely, rice germ ferment extracts-GABA treatment (10, 30, or 100 mg/kg, p.o.), especially at 100 mg/kg, normalized the sleep disturbance induced by caffeine. In locomotor tests, rice germ ferment extracts-GABA slightly but not significantly reduced the caffeine-induced increase in locomotor activity without affecting motor coordination. Additionally, rice germ ferment extracts-GABA per se did not affect the spontaneous locomotor activity and motor coordination of mice. In conclusion, rice germ ferment extracts-GABA supplementation can counter the sleep disturbance induced by caffeine, without affecting the general locomotor activities of mice.

  19. Molecular insights into how a deficiency of amylose affects carbon allocation – carbohydrate and oil analyses and gene expression profiling in the seeds of a rice waxy mutant

    Directory of Open Access Journals (Sweden)

    Zhang Ming-Zhou

    2012-12-01

    Full Text Available Abstract Background Understanding carbon partitioning in cereal seeds is of critical importance to develop cereal crops with enhanced starch yields for food security and for producing specified end-products high in amylose, β-glucan, or fructan, such as functional foods or oils for biofuel applications. Waxy mutants of cereals have a high content of amylopectin and have been well characterized. However, the allocation of carbon to other components, such as β-glucan and oils, and the regulation of the altered carbon distribution to amylopectin in a waxy mutant are poorly understood. In this study, we used a rice mutant, GM077, with a low content of amylose to gain molecular insight into how a deficiency of amylose affects carbon allocation to other end products and to amylopectin. We used carbohydrate analysis, subtractive cDNA libraries, and qPCR to identify candidate genes potentially responsible for the changes in carbon allocation in GM077 seeds. Results Carbohydrate analysis indicated that the content of amylose in GM077 seeds was significantly reduced, while that of amylopectin significantly rose as compared to the wild type BP034. The content of glucose, sucrose, total starch, cell-wall polysaccharides and oil were only slightly affected in the mutant as compared to the wild type. Suppression subtractive hybridization (SSH experiments generated 116 unigenes in the mutant on the wild-type background. Among the 116 unigenes, three, AGP, ISA1 and SUSIBA2-like, were found to be directly involved in amylopectin synthesis, indicating their possible roles in redirecting carbon flux from amylose to amylopectin. A bioinformatics analysis of the putative SUSIBA2-like binding elements in the promoter regions of the upregulated genes indicated that the SUSIBA2-like transcription factor may be instrumental in promoting the carbon reallocation from amylose to amylopectin. Conclusion Analyses of carbohydrate and oil fractions and gene expression

  20. Physiological and biochemical mechanisms of seed priming-induced chilling tolerance in rice cultivars

    Directory of Open Access Journals (Sweden)

    Saddam eHussain

    2016-02-01

    Full Text Available Rice belongs to tropical and subtropical environments and is extremely sensitive to chilling stress particularly during emergence and early stages of seedling development. Seed priming can be a good approach to enhance rice germination and stand establishment under chilling stress. The present study examined the role of different seed priming techniques viz., hydropriming, osmopriming, redox priming, chemical priming, and hormonal priming, in enhancing the chilling tolerance in rice. The most effective reagents and their pre-optimized concentrations based on preliminary experiments were used in this study. Two different rice cultivars were sown under chilling stress (18˚C and normal temperatures (28˚C in separate growth chambers. A non-primed control treatment was also maintained for comparison. Chilling stress caused erratic and delayed germination, poor seedling growth, reduced starch metabolism and lower respiration rate, while higher lipid peroxidation and hydrogen peroxide accumulation in rice seedlings of both cultivars. Nevertheless, all the seed priming treatments effectively alleviated the negative effects of chilling stress. In addition, seed priming treatments triggered the activities of superoxide dismutase, peroxidase, and catalase, and enhanced the accumulations of glutathione and free proline in rice seedlings, which suggests that these measures help prevent the rice seedlings from chilling induced oxidative stress. Chemical priming with selenium and hormonal priming with salicylic acid remained more effective treatments for both rice cultivars under chilling stress than all other priming treatments. The better performance and greater tolerance of primed rice seedlings was associated with enhanced starch metabolism, high respiration rate, lower lipid peroxidation, and strong antioxidative defense system under chilling stress.

  1. Photosynthesis Characters of a Rice Mutant, Shuanglimi%水稻突变体Shuanglimi的光合特性

    Institute of Scientific and Technical Information of China (English)

    赵帅鹏; 黄群策; 张书艮; 张时萍

    2013-01-01

    以水稻突变体Shuanglimi为试验材料,对其幼穗处于减数分裂期时剑叶的净光合速率(Pn)、蒸腾速率(Tr)、叶片气孔导度(Gs)、胞间CO2浓度(Ci)、水分利用率(WUE)和羧化效率(CE)、叶绿素含量及剑叶中维管束数目进行了测定和观察.结果表明,Shuanglimi的Pn、WUE和CE较CK分别降低了(71.18±2.08)%,(72.00±2.51)%和(79.33±3.27)%(P<0.01),而其Tr和Ci与CK之间无显著差异.Shuanglimi总叶绿素含量较CK降低,尤其是叶绿素b含量大幅降低.同时,Shuanglimi剑叶中小维管束的数目较CK降低了(18.86±1.97)%(P<0.01),而大维管束中含有3个和4个导管的特异变化数目较CK明显增加.由此可见,Shuanglimi的光合特性差异是其子粒和生殖特性差异形成的原因之一.%A new rice mutant,Shuanglimi,was selected as the experiment material.The photosynthesis of flag leaves including net photosynthesis rate (Pn),transpiration rate (Tr),stomatal conductance (Gs),intercellular CO2 concentration(Ci),water use efficiency (WUE) and carboxylation efficiency (CE) were examined,and the concentration of chlorophyll and the number of vascular bundle were also measured when the spikelet of Shuanglimi was on the meiosis stage.The results showed that Pn,WUE and CE of Shuanglimi decreased by (71.18±2.08)% (P<0.01),(72.00±2.51)% and (79.33±±3.27)% (P<0.01)compared with that of CK respectively,and T,and Ci had no significant difference.The chlorophyll content of Shuanglimi was less that that of CK,particularly,the concentration of chlorophyll b declined substantially.Meanwhile,the number of small vascular bundles in the flag leaves of Shuanglimi reduced by (18.86±1.97)% (P<0.01) compared with that of CK,and the number of big vascular bundles which included 3 or 4 vessels significantly increased.Therefore,it is the one of reasons that the differences of grain and reproduction of Shuanglimi are caused by the difference of photosynthesis characteristic.

  2. Anti-diabetic effects of rice hull smoke extract in alloxan-induced diabetic mice

    Science.gov (United States)

    We investigated the protective effect of a liquid rice hull smoke extract (RHSE) against diabetes in alloxan-induced diabetic mice. Anti-diabetic effects of RHSE were evaluated in both the rat insulinoma-1 cell line (INS-1) and diabetic ICR mice induced by inraperitoneal (ip) injection of alloxan. ...

  3. Protective effects of black rice bran against chemically-induced inflammation of mouse skin

    Science.gov (United States)

    We investigated the inhibitory effects of black rice (cv. LK1-3-6-12-1-1) bran against 12-O-tetradecanolylphorbol-13-acetate (TPA)-induced skin edema and 2,4-dinitroflurobenzene (DNFB)-induced allergic contact dermatitis (ACD) in inflammatory mouse models. We also determined the effects of the bran...

  4. Herpes simplex virus type 1 ribonucleotide reductase null mutants induce lesions in guinea pigs.

    Science.gov (United States)

    Turk, S R; Kik, N A; Birch, G M; Chiego, D J; Shipman, C

    1989-12-01

    Two herpes simplex virus type 1 ribonucleotide reductase null mutants, hrR3 and ICP6 delta, produced cutaneous lesions in guinea pigs as severe as those of wild-type strains. The lesions induced by hrR3 resulted from in vivo replication of the mutant virus, suggesting that this virus-encoded enzyme is nonessential for virus replication in guinea pigs.

  5. Beyond induced mutants: using worms to study natural variation in genetic pathways

    NARCIS (Netherlands)

    Kammenga, J.E.; Philips, P.C.; Bono, de M.; Doroszuk, A.

    2008-01-01

    Induced mutants in the nematode Caenorhabditis elegans are used to study genetic pathways of processes ranging from aging to behavior. The effects of such mutations are usually analyzed in a single wildtype background: N2. However, studies in other species demonstrate that the phenotype(s) of induce

  6. Structural changes of DNA in heavy ion-induced mutants on Arabidopsis

    Energy Technology Data Exchange (ETDEWEB)

    Tano, S.; Shikazono, N.; Tanaka, A.; Yokota, Y.; Watanabe, H. [Japan Atomic Research Research Inst., Watanuki, Takasaki (Japan). Advanced Science Research Center

    1997-09-01

    In order to investigate the frequency of structural changes induced by high LET radiation in plants, a comparison was made between DNA fragments amplified by the polymerase chain reaction (PCR) from C ion- and electron-induced Arabidopsis mutants at GL and TT loci. (orig./MG)

  7. Deficiency of antinociception and excessive grooming induced by acute immobilization stress in Per1 mutant mice.

    Science.gov (United States)

    Zhang, Jing; Wu, Zhouqiao; Zhou, Linglin; Li, Huili; Teng, Huajing; Dai, Wei; Wang, Yongqing; Sun, Zhong Sheng

    2011-01-14

    Acute stressors induce changes in numerous behavioral parameters through activation of the hypothalamic-pituitary-adrenal (HPA) axis. Several important hormones in paraventricular nucleus of the hypothalamus (PVN) play the roles in these stress-induced reactions. Corticotropin-releasing hormone (CRH), arginine-vasopressin (AVP) and corticosterone are considered as molecular markers for stress-induced grooming behavior. Oxytocin in PVN is an essential modulator for stress-induced antinociception. The clock gene, Per1, has been identified as an effecter response to the acute stresses, but its function in neuroendocrine stress systems remains unclear. In the present study we observed the alterations in grooming and nociceptive behaviors induced by acute immobilization stress in Per1 mutant mice and other genotypes (wild types and Per2 mutant). The results displayed that stress elicited a more robust effect on grooming behavior in Per1 mutant mice than in other genotypes. Subsequently, the obvious stress-induced antinociception was observed in the wild-type and Per2 mutant mice, however, in Per1 mutant, this antinociceptive effects were partially-reversed (mechanical sensitivity), or over-reversed to hyperalgesia (thermal sensitivity). The real-time qPCR results showed that in PVN, there were stress-induced up-regulations of Crh, Avp and c-fos in all of genotypes; moreover, the expression change of Crh in Per1 mutant mice was much larger than in others. Another hormonal gene, Oxt, was up-regulated induced by stress in wild-type and Per2 mutant but not in Per1 mutant. In addition, the stress significantly elevated the serum corticosterone levels without genotype-dependent differences, and accordingly the glucocorticoid receptor gene, Nr3c1, expressed with a similar pattern in PVN of all strains. Taken together, the present study indicated that in acute stress treated Per1 mutant mice, there are abnormal hormonal responses in PVN, correlating with the aberrant

  8. 水稻淡褐斑叶突变体lbsll的遗传分析与基因定位%Genetic Analysis and Gene Mapping of a Light Brown Spotted Leaf Mutant in Rice

    Institute of Scientific and Technical Information of China (English)

    奉保华; 杨杨; 施勇烽; 林璐; 陈洁; 黄奇娜; 魏彦林; Hei LEUNG; 吴建利

    2012-01-01

    A light brown spotted-leaf mutant of rice was isolated from an EMS-induced IR64 mutant bank. The mutant, designated as lbs.ll (light brown spotted-leaf 1), displayed light brown spot in the whole growth period from the first leaf to the flag leaf under natural summer field conditions. Agronomic traits including plant height, growth duration, seed-setting rate and 1000-grain weight of the mutant were significantly affected. Genetic analysis showed that the mutation was controlled by a single recessive gene, tentatively termed lbsll (t), which was mapped to the short arm of chromosome 6. With developed SSR markers, the gene was finally delimited to an interval of 130 kb between markers RM586 and RM588. The Ibsll (t) gene is likely a novel rice spotted-leaf gene since no other similar genes have been i-dentified near the chromosomal region. The genetic data and recombination populations provided will facilitate further fine-mapping and cloning of the gene.%通过EMS诱变籼稻品种IR64获得一个稳定遗传的淡褐色斑点叶突变体lbsll(light broun spotted leaf1).在自然条件下,突变体播种后10~14 d,叶片上出现淡褐色斑点,随后逐渐扩散至全叶,第1叶至剑叶上均有淡褐色斑,为全生育期性状.斑点性状的表达对株高、生育期、结实率和千粒重等农艺性状具有显著的影响.遗传分析结果表明,该淡褐色斑点叶性状受一个隐性核基因控制.将突变体lbsll与正常叶色水稻Morobereken杂交构建F2定化群体,利用SSR标记,最终将该淡褐叶基因lbsll(t)定位在第6染色体短臂上一个约130 kb的区段上.定位的结果和发展的群体为该基因的进一步精细定位和克隆奠定了基础.

  9. Temperature-sensitive mutants of Actinobacillus pleuropneumoniae induce protection in mice.

    Science.gov (United States)

    Byrd, W; Hooke, A M

    1997-01-01

    Temperature-sensitive mutants of Actinobacillus pleuropneumoniae 4074, serotype 1, were isolated after treatment with nitrosoguanidine and enrichment with penicillin and D-cycloserine. Of the four temperature-sensitive mutants evaluated in mice, one (A-1) had a tight phenotype (i.e., it ceased replication immediately after transfer to the nonpermissive temperature [37 degrees C]) and three (1-2, 4-1, and 12-1) were coasters that continued replication for up to three generations after transfer to 37 degrees C. The reversion frequencies ranged from 10(-6) to 10(-9), and cutoff temperatures ranged from 33 to 35 degrees C. No major changes were detected in the biochemical profiles; agglutination reactions; electrophoretic profiles of the lipopolysaccharides, outer membrane proteins, and hemolysin proteins; hemolytic titers; or CAMP factor reactions of the mutants and the wild-type bacteria. Groups of 3- to 5-week-old, female ICR mice were immunized intranasally with three doses of 3.5 x 10(6) CFU of the mutants over 3 weeks and subsequently challenged intranasally with 5 50% lethal doses of the parental wild-type. Protection was induced by both the tight and the coaster mutants, with the 4-1 and 12-1 coasters eliciting greater protection (67 and 82%, respectively) than that induced by the A-1 tight mutant (57%). Intranasal immunization with both phenotypes induced serum antibody responses against the surface antigens and the hemolysin protein. PMID:9169752

  10. Involvement of Jasmonate- signaling pathway in the herbivore-induced rice plant defense

    Institute of Scientific and Technical Information of China (English)

    XU Tao; ZHOU Qiang; CHEN Wei; ZHANG Guren; HE Guofeng; GU Dexiang; ZHANG Wenqing

    2003-01-01

    The expression patterns of eight defense- related genes in the herbivore-infested and jasmonate- treated (jasmonic acid, JA and its derivative MeJA) rice leaves were analyzed using RT-PCR. The results showed that Spodoptera litura Fabricius (Lepidoptera: Noctuidae) herbivory induced the expression of lipoxygenase (LOX) and allene oxide synthase (AOS) genes that are involved in the jasmonate-signaling pathway. Moreover, S. Litura damage resulted in the expression of farnesyl pyrophosphate synthase (FPS), Bowman-birk proteinase inhibitor (BBPI), phenylalanine ammonia-lyase (PAL) and other rice defense- related genes that were also induced by aqueous JA treatment or gaseous MeJA treatment. These indicated that in rice leaves, the JA-related signaling pathway was involved in the S. Litura-induced chemical defense. Mechanical damage and brown planthopper (BPH), Nilaparvata lugens (Stal) (Homoptera: Delphacidae) damage induced the expression of LOX gene, but both treatments did not induce the expression of AOS gene. However, BPH damage induced the expression of acidic pathogen-related protein 1 (PR-1a), Chitinase (PR-3), and PAL genes, which is involved in the salicylate- signaling pathway. It was suggested that salicylate-related signaling pathway or other pathways, rather than jasmonate-signaling pathway was involved in the BPH-induced rice plant defense.

  11. Morphogenesis, Anatomical Observation and Primary Genetic Analysis of a Multi-glume Floral Organ Mutant in Rice

    Institute of Scientific and Technical Information of China (English)

    PAN Cun-hong; LI Ai-hong; Wu Ru; ZHANG Ya-fang; TANG Wen; WU Chang-yin; ZHANG Qi-fa; PAN Xue-biao

    2006-01-01

    A multi-glume (mg) mutant was obtained by screening the T-DNA inserted mutant pool. Anatomical observation revealed that the florets of the mutant showed elongated leafy paleas/lemmas and palea/lemma-like structures, just like multi-glumes. Among the 215 observed florets of the mutant, 14.27% were failed to produce pistil and stamens, 23.72% showed extra floret generated on the same rachilla, while 62.01% consisted of one to nine stamens and one to three pistils in a single floret. On the other hand, in some cases the transparent bulged vesile-like tissue could be observed at the basis of filament. The mutant showed glumaceous lodicules,which prevented the florets from opening in natural conditions, while the absolute male and female sterility was an obvious character of the current mutant. Observation on the process of floral organ morphogenesis by a scanning electron microscopy (SEM) indicated that no phenotype difference in floret primordia was found between the wild-type and the mutant. Meanwhile, for the mutant, the beginning of stamen and pistil primordial differentiation was later than the wild type and the palea/lemma-like structure continued to differentiate after the formation of normal palea and lemma. Furthermore, in the mutant the asymmetrical division of floral primordial caused variation in the number of stamens and pistils. Therefore, the genetic analyses indicated that the mutation phenotype was a recessive trait controlled by a single gene and co-segregated with the T-DNA. Based on the phenotypic characteristics, it could be deduced that the mutant was the result of homeotic conversion from the function of the class E genes in ABCD model.

  12. Polypeptone induces dramatic cell lysis in ura4 deletion mutants of fission yeast.

    Directory of Open Access Journals (Sweden)

    Yuzy Matsuo

    Full Text Available Polypeptone is widely excluded from Schizosaccharomyces pombe growth medium. However, the reasons why polypeptone should be avoided have not been documented. Polypeptone dramatically induced cell lysis in the ura4 deletion mutant when cells approached the stationary growth phase, and this phenotype was suppressed by supplementation of uracil. To determine the specificity of this cell lysis phenotype, we created deletion mutants of other genes involved in de novo biosynthesis of uridine monophosphate (ura1, ura2, ura3, and ura5. Cell lysis was not observed in these gene deletion mutants. In addition, concomitant disruption of ura1, ura2, ura3, or ura5 in the ura4 deletion mutant suppressed cell lysis, indicating that cell lysis induced by polypeptone is specific to the ura4 deletion mutant. Furthermore, cell lysis was also suppressed when the gene involved in coenzyme Q biosynthesis was deleted. This is likely because Ura3 requires coenzyme Q for its activity. The ura4 deletion mutant was sensitive to zymolyase, which mainly degrades (1,3-beta-D glucan, when grown in the presence of polypeptone, and cell lysis was suppressed by the osmotic stabiliser, sorbitol. Finally, the induction of cell lysis in the ura4 deletion mutant was due to the accumulation of orotidine-5-monophosphate. Cell wall integrity was dramatically impaired in the ura4 deletion mutant when grown in the presence of polypeptone. Because ura4 is widely used as a selection marker in S. pombe, caution needs to be taken when evaluating phenotypes of ura4 mutants.

  13. silencing COI1 in rice increases susceptibility to chewing insects and impairs inducible defense.

    Directory of Open Access Journals (Sweden)

    Mao Ye

    Full Text Available The jasmonic acid (JA pathway plays a key role in plant defense responses against herbivorous insects. CORONATINE INSENSITIVE1 (COI1 is an F-box protein essential for all jasmonate responses. However, the precise defense function of COI1 in monocotyledonous plants, especially in rice (Oryza sativa L. is largely unknown. We silenced OsCOI1 in rice plants via RNA interference (RNAi to determine the role of OsCOI1 in rice defense against rice leaf folder (LF Cnaphalocrocis medinalis, a chewing insect, and brown planthopper (BPH Nilaparvata lugens, a phloem-feeding insect. In wild-type rice plants (WT, the transcripts of OsCOI1 were strongly and continuously up-regulated by LF infestation and methyl jasmonate (MeJA treatment, but not by BPH infestation. The abundance of trypsin protease inhibitor (TrypPI, and the enzymatic activities of polyphenol oxidase (PPO and peroxidase (POD were enhanced in response to both LF and BPH infestation, but the activity of lipoxygenase (LOX was only induced by LF. The RNAi lines with repressed expression of OsCOI1 showed reduced resistance against LF, but no change against BPH. Silencing OsCOI1 did not alter LF-induced LOX activity and JA content, but it led to a reduction in the TrypPI content, POD and PPO activity by 62.3%, 48.5% and 27.2%, respectively. In addition, MeJA-induced TrypPI and POD activity were reduced by 57.2% and 48.2% in OsCOI1 RNAi plants. These results suggest that OsCOI1 is an indispensable signaling component, controlling JA-regulated defense against chewing insect (LF in rice plants, and COI1 is also required for induction of TrypPI, POD and PPO in rice defense response to LF infestation.

  14. Activities of defense related enzymes induced by benzothiadiazole in rice to blast fungus

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    @@ Pretreatment of rice seedlings by foliar spraying with benzothiadiazole (BTH) could induce systematic acquired resistance (SAR) against blast (Magnaporthe grisea) and bacterial leaf blight (Xanthomonas oryzae pv. oryzae) diseases. To elucidate the physiological and biochemical mechanisms of the SAR induced by BTH, we analyzed the changes in activities of phenylalanine ammonia lyase (PAL), cinnamylalcohol dehydrogenase (CAD), peroxidase(POD), lipoxygenase(LOX),β 1,3 glucanase,and chitinase in rice seedlings of susceptible variety pretreated with BTH and challenged by M. grisea.

  15. PHOTOPERIOD-INDUCED POLLEN ABORTION IN PGMS RICE

    African Journals Online (AJOL)

    user

    2013-11-20

    Nov 20, 2013 ... nitrogenous fertilizer hence, higher yield (Hu, 1993). Later came the high yielding .... Rice lines ZAU11S106 and control ZAU11F121 were sown on May. 14th in nine rows each ... release the pollen cells. Anther-husks were ...

  16. Antibodies with higher bactericidal activity induced by a Neisseria gonorrhoeae Rmp deletion mutant strain.

    Directory of Open Access Journals (Sweden)

    Guocai Li

    Full Text Available Neisseria gonorrhoeae (N. gonorrhoeae outer membrane protein reduction modifiable protein (Rmp has strong immunogenicity. However, anti-Rmp antibodies block rather than preserve the antibacterial effects of protective antibodies, which hampers the development of vaccines for gonococcal infections. We herein constructed an Rmp deletion mutant strain of N. gonorrhoeae by gene homologous recombination. The 261-460 nucleotide residues of Rmp gene amplified from N. gonorrhoeae WHO-A strain were replaced with a kanamycin-resistant Kan gene amplified from pET-28a. The resultant hybridized DNA was transformed into N. gonorrhoeae WHO-A strain. PCR was used to screen the colonies in which wild-type Rmp gene was replaced with a mutant gene fragment. Western blotting revealed that the Rmp deletion mutant strain did not express Rmp protein. Rmp deletion did not alter the morphological and Gram staining properties of the mutant strain that grew slightly more slowly than the wild-type one. Rmp gene mutated stably throughout 25 generations of passage. Antibody-mediated complement-dependent cytotoxicity assay indicated that the antibodies induced by the mutant strain had evidently higher bactericidal activities than those induced by the wild-type strain. Further modification of the Rmp deletion mutant strain is still required in the development of novel live attenuated vaccines for gonorrhea by Opa genes deletion or screening of phenotypic variant strains that do not express Opa proteins.

  17. Antibodies with higher bactericidal activity induced by a Neisseria gonorrhoeae Rmp deletion mutant strain.

    Science.gov (United States)

    Li, Guocai; Xie, Rushan; Zhu, Xiaoping; Mao, Yanli; Liu, Shuangxi; Jiao, Hongmei; Yan, Hua; Xiong, Kun; Ji, Mingchun

    2014-01-01

    Neisseria gonorrhoeae (N. gonorrhoeae) outer membrane protein reduction modifiable protein (Rmp) has strong immunogenicity. However, anti-Rmp antibodies block rather than preserve the antibacterial effects of protective antibodies, which hampers the development of vaccines for gonococcal infections. We herein constructed an Rmp deletion mutant strain of N. gonorrhoeae by gene homologous recombination. The 261-460 nucleotide residues of Rmp gene amplified from N. gonorrhoeae WHO-A strain were replaced with a kanamycin-resistant Kan gene amplified from pET-28a. The resultant hybridized DNA was transformed into N. gonorrhoeae WHO-A strain. PCR was used to screen the colonies in which wild-type Rmp gene was replaced with a mutant gene fragment. Western blotting revealed that the Rmp deletion mutant strain did not express Rmp protein. Rmp deletion did not alter the morphological and Gram staining properties of the mutant strain that grew slightly more slowly than the wild-type one. Rmp gene mutated stably throughout 25 generations of passage. Antibody-mediated complement-dependent cytotoxicity assay indicated that the antibodies induced by the mutant strain had evidently higher bactericidal activities than those induced by the wild-type strain. Further modification of the Rmp deletion mutant strain is still required in the development of novel live attenuated vaccines for gonorrhea by Opa genes deletion or screening of phenotypic variant strains that do not express Opa proteins.

  18. A Mutant-p53/Smad complex opposes p63 to empower TGFbeta-induced metastasis.

    Science.gov (United States)

    Adorno, Maddalena; Cordenonsi, Michelangelo; Montagner, Marco; Dupont, Sirio; Wong, Christine; Hann, Byron; Solari, Aldo; Bobisse, Sara; Rondina, Maria Beatrice; Guzzardo, Vincenza; Parenti, Anna R; Rosato, Antonio; Bicciato, Silvio; Balmain, Allan; Piccolo, Stefano

    2009-04-03

    TGFbeta ligands act as tumor suppressors in early stage tumors but are paradoxically diverted into potent prometastatic factors in advanced cancers. The molecular nature of this switch remains enigmatic. Here, we show that TGFbeta-dependent cell migration, invasion and metastasis are empowered by mutant-p53 and opposed by p63. Mechanistically, TGFbeta acts in concert with oncogenic Ras and mutant-p53 to induce the assembly of a mutant-p53/p63 protein complex in which Smads serve as essential platforms. Within this ternary complex, p63 functions are antagonized. Downstream of p63, we identified two candidate metastasis suppressor genes associated with metastasis risk in a large cohort of breast cancer patients. Thus, two common oncogenic lesions, mutant-p53 and Ras, selected in early neoplasms to promote growth and survival, also prefigure a cellular set-up with particular metastasis proclivity by TGFbeta-dependent inhibition of p63 function.

  19. Silicon-induced reversibility of cadmium toxicity in rice

    Science.gov (United States)

    Farooq, Muhammad Ansar; Detterbeck, Amelie; Clemens, Stephan; Dietz, Karl-Josef

    2016-01-01

    Silicon (Si) modulates tolerance to abiotic stresses, but little is known about the reversibility of stress effects by supplementing previously stressed plants with Si. This is surprising since recovery experiments might allow mechanisms of Si-mediated amelioration to be addressed. Rice was exposed to 10 µM CdCl2 for 4 d in hydroponics, followed by 0.6mM Si(OH)4 supplementation for 4 d. Si reversed the effects of Cd, as reflected in plant growth, photosynthesis, elemental composition, and some biochemical parameters. Cd-dependent deregulation of nutrient homeostasis was partially reversed by Si supply. Photosynthetic recovery within 48h following Si supply, coupled with strong stimulation of the ascorbate–glutathione system, indicates efficient activation of defense. The response was further verified by transcript analyses with emphasis on genes encoding members of the stress-associated protein (SAP) family. The transcriptional response to Cd was mostly reversed following Si supply. Reprogramming of the Cd response was obvious for Phytochelatin synthase 1, SAP1 , SAP14, and the transcription factor genes AP2/Erf020, Hsf31, and NAC6 whose transcript levels were strongly activated in roots of Cd-stressed rice, but down-regulated in the presence of Si. These findings, together with changes in biochemical parameters, highlight the significance of Si in growth recovery of Cd-stressed rice and indicate a decisive role for readjusting cell redox homeostasis. PMID:27122572

  20. A conditionally lethal mutant of Salmonella Typhimurium induces a protective response in mice.

    Science.gov (United States)

    Hidalgo, Alejandro A; Villagra, Nicolás A; Jerez, Sebastián A; Fuentes, Juan A; Mora, Guido C

    2016-02-01

    Here we present the design of a conditionally lethal mutant of Salmonella enterica serovar Typhimurium (S. Typhimurium) which growth depends on tetracycline (Tet). Four mutants of S. Typhimurium, with Tet-conditional growth, were created by inserting the tetRA cassette. Three of the mutants presented a conditional-lethal phenotype in vitro. One mutant in the yabB gene remained conditional inside cells and did not persisted after 24 h in cell cultures. The capacity of S. Typhimurium yabB::tetRA to invade deep organs was investigated in intraperitoneally (IP) infected mice fed with or without chlortetracycline (CTet), a Tet analog with lower antibiotic activity. The yabB::tetRA mutant was undetectable in liver or spleen of animals under normal diet, while in mice under diet including CTet, yabB::tetRA invaded at a level comparable to the WT in mice under normal diet. Moreover, yabB::tetRA produced a strong humoral-immunoresponse after one IP immunization with 10(6) bacteria, measured as serum reactivity against S. Typhimurium whole cell extract. By contrast, oral immunization with 10(6) bacteria was weaker and variable on inducing antibodies. Consistently, IP infected mice were fully protected in a challenge with 10(4) oral S. Typhimurium, while protection was partial in orally immunized mice. Our data indicate that S. Typhimurium yabB::tetRA is a conditionally attenuated strain capable of inducing a protective response in mice in non-permissive conditions.

  1. Gravity-Induced Modifications to Development in Hypocotyls of Arabidopsis Tubulin Mutants1[W][OA

    Science.gov (United States)

    Matsumoto, Shouhei; Kumasaki, Saori; Soga, Kouichi; Wakabayashi, Kazuyuki; Hashimoto, Takashi; Hoson, Takayuki

    2010-01-01

    We investigated the roles of cortical microtubules in gravity-induced modifications to the development of stem organs by analyzing morphology and orientation of cortical microtubule arrays in hypocotyls of Arabidopsis (Arabidopsis thaliana) tubulin mutants, tua3(D205N), tua4(S178Δ), and tua6(A281T), cultivated under 1g and hypergravity (300g) conditions. Hypocotyls of tubulin mutants were shorter and thicker than the wild type even at 1g, and hypergravity further suppressed elongation and stimulated expansion. The degree of such changes was clearly smaller in tubulin mutants, in particular in tua6. Hypocotyls of tubulin mutants also showed either left-handed or right-handed helical growth at 1g, and the degree of twisting phenotype was intensified under hypergravity conditions, especially in tua6. Hypergravity induced reorientation of cortical microtubules from transverse to longitudinal directions in epidermal cells of wild-type hypocotyls. In tubulin mutants, especially in tua6, the percentage of cells with longitudinal microtubules was high even at 1g, and it was further increased by hypergravity. The twisting phenotype was most obvious at cells 10 to 12 from the top, where reorientation of cortical microtubules from transverse to longitudinal directions occurred. Moreover, the left-handed helical growth mutants (tua3 and tua4) had right-handed microtubule arrays, whereas the right-handed mutant (tua6) had left-handed arrays. There was a close correlation between the alignment angle of epidermal cell files and the alignment of cortical microtubules. Gadolinium ions, blockers of mechanosensitive ion channels (mechanoreceptors), suppressed the twisting phenotype in tubulin mutants under both 1g and 300g conditions. Microtubule arrays in tubulin mutants were oriented more transversely by gadolinium treatment, irrespective of gravity conditions. These results support the hypothesis that cortical microtubules play an essential role in maintenance of normal growth

  2. Gravity-induced modifications to development in hypocotyls of Arabidopsis tubulin mutants.

    Science.gov (United States)

    Matsumoto, Shouhei; Kumasaki, Saori; Soga, Kouichi; Wakabayashi, Kazuyuki; Hashimoto, Takashi; Hoson, Takayuki

    2010-02-01

    We investigated the roles of cortical microtubules in gravity-induced modifications to the development of stem organs by analyzing morphology and orientation of cortical microtubule arrays in hypocotyls of Arabidopsis (Arabidopsis thaliana) tubulin mutants, tua3(D205N), tua4(S178Delta), and tua6(A281T), cultivated under 1g and hypergravity (300g) conditions. Hypocotyls of tubulin mutants were shorter and thicker than the wild type even at 1g, and hypergravity further suppressed elongation and stimulated expansion. The degree of such changes was clearly smaller in tubulin mutants, in particular in tua6. Hypocotyls of tubulin mutants also showed either left-handed or right-handed helical growth at 1g, and the degree of twisting phenotype was intensified under hypergravity conditions, especially in tua6. Hypergravity induced reorientation of cortical microtubules from transverse to longitudinal directions in epidermal cells of wild-type hypocotyls. In tubulin mutants, especially in tua6, the percentage of cells with longitudinal microtubules was high even at 1g, and it was further increased by hypergravity. The twisting phenotype was most obvious at cells 10 to 12 from the top, where reorientation of cortical microtubules from transverse to longitudinal directions occurred. Moreover, the left-handed helical growth mutants (tua3 and tua4) had right-handed microtubule arrays, whereas the right-handed mutant (tua6) had left-handed arrays. There was a close correlation between the alignment angle of epidermal cell files and the alignment of cortical microtubules. Gadolinium ions, blockers of mechanosensitive ion channels (mechanoreceptors), suppressed the twisting phenotype in tubulin mutants under both 1g and 300 g conditions. Microtubule arrays in tubulin mutants were oriented more transversely by gadolinium treatment, irrespective of gravity conditions. These results support the hypothesis that cortical microtubules play an essential role in maintenance of normal

  3. Analysis of Chloroplast Ultrastructure, Photosystem Ⅱ Light Harvesting Complexes and Chlorophyll Synthesis in a Chlorophyll-Less Rice Mutant W2555

    Institute of Scientific and Technical Information of China (English)

    XU Pei-zhou; LI Yun; YUAN Shu; ZHANG Hong-yu; WANG Xu-dong; LIN Hong-hui; WU Xian-jun

    2006-01-01

    A comparative study on chloroplast ultrastructure and light harvesting complex of photosystem Ⅱ (LHC Ⅱ) was conducted between a new rice mutant (W2555) and its wild type (WT). The chloroplasts of W2555 had less thylakoids and grana stacks compared with the wild type. There was no significant change in the composition of LHC Ⅱ polypeptide in W2555, while a decline had been noted in LHC Ⅱ content. Northern blot analysis with a specific cab gene probe showed no appreciable difference in the LHC Ⅱ mRNA level between the W2555 and its wild type. The precursors of chlorophyll synthesis, δ-aminolevulinic acid (ALA)and porphobilinogen (PBG) were over accumulated in W2555, but the other precursors were all decreased. These results indicated that the decreased level of LHC Ⅱ in the mutant W2555 was attributed to the change of cab gene transcription, but a blockage in chlorophyll biosynthesis due to the formation of uroporphyrinogen Ⅲ (Urogen Ⅲ).

  4. Decreased photosynthesis in the erect panicle 3 (ep3) mutant of rice is associated with reduced stomatal conductance and attenuated guard cell development.

    Science.gov (United States)

    Yu, Hongyang; Murchie, Erik H; González-Carranza, Zinnia H; Pyke, Kevin A; Roberts, Jeremy A

    2015-03-01

    The ERECT PANICLE 3 gene of rice encodes a peptide that exhibits more than 50% sequence identity with the Arabidopsis F-box protein HAWAIIAN SKIRT (HWS). Ectopic expression of the Os02g15950 coding sequence, driven by the HWS (At3g61950) promoter, rescued the hws-1 flower phenotype in Arabidopsis confirming that EP3 is a functional orthologue of HWS. In addition to displaying an erect inflorescence phenotype, loss-of-function mutants of Os02g15950 exhibited a decrease in leaf photosynthetic capacity and stomatal conductance. Analysis of a range of physiological and anatomical features related to leaf photosynthesis revealed no alteration in Rubisco content and no notable changes in mesophyll size or arrangement. However, both ep3 mutant plants and transgenic lines that have a T-DNA insertion within the Os02g15950 (EP3) gene exhibit smaller stomatal guard cells compared with their wild-type controls. This anatomical characteristic may account for the observed decrease in leaf photosynthesis and provides evidence that EP3 plays a role in regulating stomatal guard cell development.

  5. An S-domain receptor-like kinase, OsSIK2, confers abiotic stress tolerance and delays dark-induced leaf senescence in rice.

    Science.gov (United States)

    Chen, Li-Juan; Wuriyanghan, Hada; Zhang, Yu-Qin; Duan, Kai-Xuan; Chen, Hao-Wei; Li, Qing-Tian; Lu, Xiang; He, Si-Jie; Ma, Biao; Zhang, Wan-Ke; Lin, Qing; Chen, Shou-Yi; Zhang, Jin-Song

    2013-12-01

    Receptor-like kinases play important roles in plant development and defense responses; however, their functions in other processes remain unclear. Here, we report that OsSIK2, an S-domain receptor-like kinase from rice (Oryza sativa), is involved in abiotic stress and the senescence process. OsSIK2 is a plasma membrane-localized protein with kinase activity in the presence of Mn(2+). OsSIK2 is expressed mainly in rice leaf and sheath and can be induced by NaCl, drought, cold, dark, and abscisic acid treatment. Transgenic plants overexpressing OsSIK2 and mutant sik2 exhibit enhanced and reduced tolerance to salt and drought stress, respectively, compared with the controls. Interestingly, a truncated version of OsSIK2 without most of the extracellular region confers higher salt tolerance than the full-length OsSIK2, likely through the activation of different sets of downstream genes. Moreover, seedlings of OsSIK2-overexpressing transgenic plants exhibit early leaf development and a delayed dark-induced senescence phenotype, while mutant sik2 shows the opposite phenotype. The downstream PR-related genes specifically up-regulated by full-length OsSIK2 or the DREB-like genes solely enhanced by truncated OsSIK2 are all induced by salt, drought, and dark treatments. These results indicate that OsSIK2 may integrate stress signals into a developmental program for better adaptive growth under unfavorable conditions. Manipulation of OsSIK2 should facilitate the improvement of production in rice and other crops.

  6. Progranulin is neurotrophic in vivo and protects against a mutant TDP-43 induced axonopathy.

    Directory of Open Access Journals (Sweden)

    Angela S Laird

    Full Text Available Mislocalization, aberrant processing and aggregation of TAR DNA-binding protein 43 (TDP-43 is found in the neurons affected by two related diseases, amyotrophic lateral sclerosis (ALS and frontotemporal lobe dementia (FTLD. These TDP-43 abnormalities are seen when TDP-43 is mutated, such as in familial ALS, but also in FTLD, caused by null mutations in the progranulin gene. They are also found in many patients with sporadic ALS and FTLD, conditions in which only wild type TDP-43 is present. The common pathological hallmarks and symptomatic cross over between the two diseases suggest that TDP-43 and progranulin may be mechanistically linked. In this study we aimed to address this link by establishing whether overexpression of mutant TDP-43 or knock-down of progranulin in zebrafish embryos results in motor neuron phenotypes and whether human progranulin is neuroprotective against such phenotypes. Mutant TDP-43 (A315T mutation induced a motor axonopathy characterized by short axonal outgrowth and aberrant branching, similar, but more severe, than that induced by mutant SOD1. Knockdown of the two zebrafish progranulin genes, grna and grnb, produced a substantial decrease in axonal length, with knockdown of grna alone producing a greater decrease in axonal length than grnb. Progranulin overexpression rescued the axonopathy induced by progranulin knockdown. Interestingly, progranulin also rescued the mutant TDP-43 induced axonopathy, whilst it failed to affect the mutant SOD1-induced phenotype. TDP-43 was found to be nuclear in all conditions described. The findings described here demonstrate that progranulin is neuroprotective in vivo and may have therapeutic potential for at least some forms of motor neuron degeneration.

  7. Cutin monomer induces expression of the rice OsLTP5 lipid transfer protein gene.

    Science.gov (United States)

    Kim, Tae Hyun; Park, Jong Ho; Kim, Moon Chul; Cho, Sung Ho

    2008-01-01

    Treatment with the cutin monomer 16-hydroxypalmitic acid (HPA), a major component of cutin, elicited the synthesis of hydrogen peroxide (H2O2) in rice leaves and induced the expression of the lipid transfer protein gene OsLTP5. Treatment with HPA also induced expression of OsLTP1, OsLTP2, and the pathogen-related PR-10 genes to a lesser extent. The OsLTP5 transcript was expressed prominently in stems and flowers, but was barely detectable in leaves. Expression of OsLTP5 was induced in shoots in response to ABA and salicylic acid. It is proposed that HPA is perceived by rice as a signal, inducing defense reactions.

  8. Induced protein polymorphisms and nutritional quality of gamma irradiation mutants of sorghum

    Energy Technology Data Exchange (ETDEWEB)

    Mehlo, Luke, E-mail: LMehlo@csir.co.za [CSIR Biosciences, Meiring Naude Road, P.O. Box 395, Pretoria 0001 (South Africa); Mbambo, Zodwa [CSIR Biosciences, Meiring Naude Road, P.O. Box 395, Pretoria 0001 (South Africa); Microbiology Discipline, School of Life Sciences, University of KwaZulu-Natal (Westville Campus), Private Bag X54001, Durban 4000 (South Africa); Bado, Souleymane [Plant Breeding and Genetics Laboratory – Joint FAO/IAEA Agriculture and Biotechnology Laboratory, International Atomic Energy Agency Laboratories, A-2444 Seibersdorf (Austria); Lin, Johnson [Microbiology Discipline, School of Life Sciences, University of KwaZulu-Natal (Westville Campus), Private Bag X54001, Durban 4000 (South Africa); Moagi, Sydwell M.; Buthelezi, Sindisiwe; Stoychev, Stoyan; Chikwamba, Rachel [CSIR Biosciences, Meiring Naude Road, P.O. Box 395, Pretoria 0001 (South Africa)

    2013-09-15

    Highlights: • We analyse kafirin protein polymorphisms induced by gamma irradiation in sorghum. • One mutant with suppressed kafirins in the endosperm accumulated them in the germ. • Kafirin polymorphisms were associated with high levels of free amino acids. • Nutritional value of sorghum can be improved significantly by induced mutations. - Abstract: Physical and biochemical analysis of protein polymorphisms in seed storage proteins of a mutant population of sorghum revealed a mutant with redirected accumulation of kafirin proteins in the germ. The change in storage proteins was accompanied by an unusually high level accumulation of free lysine and other essential amino acids in the endosperm. This mutant further displayed a significant suppression in the synthesis and accumulation of the 27 kDa γ-, 24 kDa α-A1 and the 22 kDa α-A2 kafirins in the endosperm. The suppression of kafirins was counteracted by an upsurge in the synthesis and accumulation of albumins, globulins and other proteins. The data collectively suggest that sorghum has huge genetic potential for nutritional biofortification and that induced mutations can be used as an effective tool in achieving premium nutrition in staple cereals.

  9. 两个水稻叶色突变体的光合特性研究%Characteristics of Photosynthesis in Two Leaf Color Mutants of Rice

    Institute of Scientific and Technical Information of China (English)

    吕典华; 宗学凤; 王三根; 凌英华; 桑贤春; 何光华

    2009-01-01

    Two leaf color mutants, ygl5 and pygl1, were identified from the progeny of Jinhui 10 treated with EMS, and Jinhui 10 was an excellent restorer line bred in the Rice Research Institute of Southwest University. This paper reported their photosynthetic parameters and ultra-structure of chloroplast. Compared with the original parent, both mutants had a decline content of chlorophylls significantly, especially in total chlorophylls for ygl5 and in chlorophyll b for the pygl1. Photosynthetic and chlorophyll fluorescence parameters were determined by L1-6400, the results showed that the net photosynthetic rate (P_n), transpiration rate (T_r) and stomatal conductance (G_s) were higher in the mutants, with higher light compensation point (LCP) and lower light saturation point (LSP) as well as lower CO_2 compensation point (CCP) and CO_2 saturation point (CSP). Compared with Jinhui 10, the mutants displayed less chloroplast grana and higher optional maximal photochemical efficiency of PS II (F_v/F_m)> actual photochemical efficiency of PSII (Φ_(PSII)), and photochemical quenching (q_P), suggesting that the mutants have higher light energy capture and conversion efficiencies as well as the electron transport efficiency.%以恢复系缙恢10号为对照,对它的两个叶色突变体ygl5和pygl1在孕穗期的光合、荧光特性和叶绿体超微结构进行了研究.结果表明,两个突变体叶绿体内基粒数量明显少于对照,叶绿素含量也大幅下降,其中ygl5表现为叶绿索总体缺乏,pygl1表现为叶绿素b严重缺乏;ygl5和pygl1的P_n、光饱和点、光补偿点、暗呼吸速率、表观量子效率和羧化效率都显著高于对照,而CO_2饱和点、CO_2补偿点和光呼吸速率则低于对照;其F_v/F_m、Φ_(PSII)和q_P均显著高于对照,表明突变体ygl5和pygl1具有较高的光能捕获效率和转换效率.

  10. Physiological and biochemical characteristics of laboratory induced mutants of Botrytis cinerea with resistance to fluazinam.

    Science.gov (United States)

    Shao, Wenyong; Zhang, Yu; Ren, Weichao; Chen, Changjun

    2015-01-01

    Botrytis cinerea is a necrotrophic and filamentous fungus with a high risk of developing resistance to fungicides. The pyridinamine fungicide fluazinam has been reported to have excellent activity against B. cinerea and better effect on controlling gray mold. In this study, the physiological and biochemical characteristics of laboratory-induced mutants of B. cinerea with resistance to fluazinam has been investigated. Compared to the wild-type strains, the fluazinam-resistant mutants had a significant decrease in respiratory rate, glycerol, oxalate, and ATP contents, and an increase in ATPase activity and sensitivity to osmotic pressure, but did not differ in cell membrane permeability. Sequencing indicated that two parental strains and four resistant mutants were identical in the nucleotide sequence of F-ATPase gene. These results will enrich our understanding of the resistance mechanism of B. cinerea to fluazinam.

  11. Screening of gamma radiation-induced pathogen resistance rice lines against Xanthomonas oryzae pv. oryzae

    Energy Technology Data Exchange (ETDEWEB)

    Lim, Chan Ju; Lee, Ha Yeon; Kim, Woong Bom; Ahmad, Raza; Moon, Jae Sun; Kwon, Suk Yoon [Korea Research Institute of Beoscience and Biotechnology, Daejeon (Korea, Republic of); Kim, Dong Sub [Korea Atomic Energy Research Institute, Jeongeup (Korea, Republic of)

    2010-09-15

    Bacterial blight is one of the most serious diseases of rice (Oryza sativa L.), and it has been known that Xanthomonas oryzae pv. oryzae (Xoo) causes this disease symptom. To develop resistance rice cultivars against Xoo, 3,000 lines of M{sub 3}, which were irradiated with gamma ray, were tested by 'scissor-dip method' primarily, and 191 putative resistant lines were selected. In M{sub 4} generation, these lines were screened again with various ways such as measuring of symptom of bacterial blight in leaf, number of tiller, fresh weight, and phenotypic segregation ratio in next generation. Finally, six resistance lines were selected. RT-PCR analysis revealed that these lines displayed high level of R-genes such as Xa21, Pi36, and Pi-ta. These results indicate that mutations by gamma ray cause disruptions of regulatory signal transduction systems of these R-genes. Furthermore, these selected mutants could be useful for the development of rice cultivar resistant to Xoo.

  12. Identification of Francisella novicida mutants that fail to induce prostaglandin E2 synthesis by infected macrophages.

    Directory of Open Access Journals (Sweden)

    Matthew Dale Woolard

    2013-02-01

    Full Text Available Francisella tularensis is the causative agent of tularemia. We have previously shown that infection with F. tularensis Live Vaccine Strain (LVS induces macrophages to synthesize prostaglandin E2 (PGE2. Synthesis of PGE2 by F. tularensis infected macrophages results in decreased T cell proliferation in vitro and increased bacterial survival in vivo. Although we understand some of the biological consequences of F. tularensis induced PGE2 synthesis by macrophages, we do not understand the cellular pathways (neither host nor bacterial that result in up-regulation of the PGE2 biosynthetic pathway in F. tularensis infected macrophages. We took a genetic approach to begin to understand the molecular mechanisms of bacterial induction of PGE2 synthesis from infected macrophages. To identify F. tularensis genes necessary for the induction of PGE2 in primary macrophages, we infected cells with individual mutants from the closely related strain Francisella tularensis subspecies novicida U112 (U112 two allele mutant library. Twenty genes were identified that when disrupted resulted in U112 mutant strains unable to induce the synthesis of PGE2 by infected macrophages. Fourteen of the genes identified are located within the Francisella pathogenicity island (FPI. Genes in the FPI are required for F. tularensis to escape from the phagosome and replicate in the cytosol, which might account for the failure of U112 with transposon insertions within the FPI to induce PGE2. This implies that U112 mutant strains that do not grow intracellularly would also not induce PGE2. We found that U112 clpB::Tn grows within macrophages yet fails to induce PGE2, while U112 pdpA::Tn does not grow yet does induce PGE2. We also found that U112 iglC::Tn neither grows nor induces PGE2. These findings indicate that there is dissociation between intracellular growth and the ability of F. tularensis to induce PGE2 synthesis. These mutants provide a critical entrée into the pathways used

  13. Identification and Gene Mapping of a Spotted-leaf Mutant hm197 in Rice%水稻斑点叶突变体hm197的鉴定及其基因定位

    Institute of Scientific and Technical Information of China (English)

    李小红; 吴建利; 施勇烽; 张晓波; 奉保华; 宋莉欣; 王惠梅; 徐霞; 黄奇娜; 郭丹

    2015-01-01

    A stable inherited rice spotted-leaf mutant hm197 was isolated from a diepoxybutane-induced IR64 mutant bank.Under natural conditions,brown lesions were first observed on the tips of the mutant leaves 10 weeks after sowing,and spread gradually downward to cover the whole leaf blades.Genetic analysis indicated that the mutant trait was controlled by a novel single recessive nuclear gene,tentatively termed as splhm97 ,located in the long arm of chromosome 4.Agronomic traits including the plant height,seed-setting rate and 1000-grain weight were significantly decreased in hm197 .The initiation of brown lesions in hm197 was induced by natural sun light.In addition,the photosynthetic pigment contents and net photosynthetic rate in the mutant were significantly lower than those in the wild type IR64.Histochemical analysis showed that H2 O2 and O2∸ accumulation were presented in and around the lesions in hm197 .Furthermore,senescence-related parameters including the activities of SOD,APX,total soluble protein and MDA contents were also significantly altered in the mutant while resistance to bacterial blight pathogens was largely enhanced in the mutant in contrast to the wild type IR64.%通过双环氧丁烷(diepoxybutane)诱变籼稻品种 IR64获得遗传稳定的水稻褐色斑点叶突变体hm197.在自然条件下,该突变体褐色斑点自播种后10周开始于叶尖出现,而后慢慢扩散至全叶.遗传分析表明,该褐色斑点性状受一对隐性核基因控制,暂名splhm197,并将其定位在第4染色体长臂上140 kb的区段内.与野生型IR64相比,突变体株高、结实率和千粒重等农艺性状均显著下降.遮光处理表明,hm197褐色斑点的形成受自然光照的诱导.此外,hm197光合色素含量和光合效率也比野生型显著降低.组织化学分析表明,突变体中有过氧化氢和大量超氧阴离子 O2∸的沉积.与 IR64相比, hm197叶片中清除氧自由基酶系统中 SOD和 APX活性极显著上升,其余

  14. Stability Test For Sorghum Mutant Lines Derived From Induced Mutations with Gamma-Ray Irradiation

    Directory of Open Access Journals (Sweden)

    S. Human

    2011-12-01

    Full Text Available Sorghum breeding program had been conducted at the Center for the Application of Isotopes and Radiation Technology, BATAN. Plant genetic variability was increased through induced mutations using gamma-ray irradiation. Through selection process in successive generations, some promising mutant lines had been identified to have good agronomic characteristics with high grain yield. These breeding lines were tested in multi location trials and information of the genotypic stability was obtained to meet the requirements for officially varietal release by the Ministry of Agriculture. A total of 11 sorghum lines and varieties consisting of 8 mutant lines derived from induced mutations (B-100, B-95, B-92, B-83, B-76, B-75, B-69 and Zh-30 and 3 control varieties (Durra, UPCA-S1 and Mandau were included in the experiment. All materials were grown in 10 agro-ecologically different locations namely Gunungkidul, Bantul, Citayam, Garut, Lampung, Bogor, Anyer, Karawaci, Cianjur and Subang. In each location, the local adaptability test was conducted by randomized block design with 3 replications. Data of grain yield was used for evaluating genotypic stability using AMMI approach. Results revealed that sorghum mutation breeding had generated 3 mutant lines (B-100, B-76 and Zh-30 exhibiting grain yield significantly higher than the control varieties. These mutant lines were genetically stable in all locations so that they would be recommended for official release as new sorghum varieties to the Ministry of Agriculture

  15. Proton induced X-ray emission and electron microscopy analysis of induced mutants of sorghum

    CSIR Research Space (South Africa)

    Mbambo, Z

    2014-01-01

    Full Text Available of elements in preferential accumulation tissues and entire changes in cellular localization. Transmission and scanning electron microscopy of the mutants resolved changes in size, shape, ultra-structure and packed cell volumes of protein- and starch bodies...

  16. Association of specific pectin methylesterases with Al-induced root elongation inhibition in rice.

    Science.gov (United States)

    Yang, Xiao Ying; Zeng, Zhang Hui; Yan, Jing Ying; Fan, Wei; Bian, Hong Wu; Zhu, Mu Yuan; Yang, Jian Li; Zheng, Shao Jian

    2013-08-01

    The negative charges of cell wall pectin molecules attributed by pectin methylesterase (PME, EC 3.1.1.11) contribute to Al binding capacity. We examined the expression profiles of 35 members of the PME gene family in the root apex of an Al-sensitive rice 'Zhefu802' under Al stress. While root elongation was inhibited by 40% after 3-h exposure to 25 µM Al, cell wall PME activity and the abundance of eight PME genes transcripts were increased. The same Al treatment which had almost no effect on root elongation of an Al-resistant rice ssp. japonica 'Nipponbare' did not change the expression patterns of these eight PME genes. However, when Al concentration was increased to 50 µM, by which the root elongation of 'Nipponbare' was inhibited by 40% too, the expression of these PME genes were also upregulated except two genes with no signal. These suggest a possible correlation between the upregulated genes and Al-induced inhibition of root elongation in rice. Furthermore, these eight PME genes behaved differently when subjected to CdCl2 and LaCl3 treatments, implying the specificity of different PME genes in response to different metal toxicities. The transgenic rice overexpressing one of these eight PME genes OsPME14 showed higher PME activity and Al content in root tip cell wall, and became more sensitive to Al stress, verifying the involvement of the specific PME gene in Al toxicity. Therefore, our results provided the molecular evidence to connect the expression of specific PME genes with the Al-induced inhibition of root elongation in rice.

  17. Ligand-induced conformational changes in wild-type and mutant yeast pyruvate kinase.

    Science.gov (United States)

    Collins, R A; Kelly, S M; Price, N C; Fothergill-Gilmore, L A; Muirhead, H

    1996-12-01

    A mutant form of pyruvate kinase in which serine 384 has been mutated to proline has been engineered in the yeast Saccharomyces cerevisiae. Residue 384 is located in a helix in a subunit interface of the tetrameric enzyme, and the mutation was anticipated to alter the conformation of the helix and hence destabilize the interface. Previous results indicate that the mutant favours the T quaternary conformation over the R conformation, and this is confirmed by the results presented here. Addition of phosphoenol-pyruvate (PEP), ADP and fructose-1, 6-bisphosphate (Fru-1.6-P2) singly to the wild-type and mutant enzymes results in a significant quenching of tryptophan fluorescence (12-44%), and for Fru-1,6-P2, a red shift of 15 nm in the emission maximum. Fluorescence titration experiments showed that PEP, ADP and Fru-1,6-P2 induce conformations which have similar ligand-binding properties in the wild-type and mutant enzymes. However, the Fru-1,6-P2 induced conformation is demonstrably different from those induced by either ADP or PEP. The enzymes differ in their susceptibility to trypsin digestion and N-ethylmaleimide inhibition. The thermal stability of the enzyme is unaltered by the mutation. Far-UV CD spectra show that both enzymes adopt a similar overall secondary structure in solution. Taken together, the results suggest that the Ser384-Pro mutation causes the enzyme to adopt a different tertiary and/or quaternary structure from the wild-type enzyme and affects the type and extent of the conformational changes induced in the enzyme upon ligand binding. A simplified minimal reaction mechanism is proposed in which the R and T states differ in both affinity and kcat. Thus, in terms of the models of cooperativity and allosteric interaction, pyruvate kinase is both a K and a V system.

  18. ABA Inducible Rice Protein Phosphatase 2C Confers ABA Insensitivity and Abiotic Stress Tolerance in Arabidopsis

    Science.gov (United States)

    Singh, Amarjeet; Jha, Saroj K.; Bagri, Jayram; Pandey, Girdhar K.

    2015-01-01

    Arabidopsis PP2C belonging to group A have been extensively worked out and known to negatively regulate ABA signaling. However, rice (Oryza sativa) orthologs of Arabidopsis group A PP2C are scarcely characterized functionally. We have identified a group A PP2C from rice (OsPP108), which is highly inducible under ABA, salt and drought stresses and localized predominantly in the nucleus. Genetic analysis revealed that Arabidopsis plants overexpressing OsPP108 are highly insensitive to ABA and tolerant to high salt and mannitol stresses during seed germination, root growth and overall seedling growth. At adult stage, OsPP108 overexpression leads to high tolerance to salt, mannitol and drought stresses with far better physiological parameters such as water loss, fresh weight, chlorophyll content and photosynthetic potential (Fv/Fm) in transgenic Arabidopsis plants. Expression profile of various stress marker genes in OsPP108 overexpressing plants revealed interplay of ABA dependent and independent pathway for abiotic stress tolerance. Overall, this study has identified a potential rice group A PP2C, which regulates ABA signaling negatively and abiotic stress signaling positively. Transgenic rice plants overexpressing this gene might provide an answer to the problem of low crop yield and productivity during adverse environmental conditions. PMID:25886365

  19. Characterization and Gene Fine Mapping of a Rice Dominant Spotted-leaf Mutant%一个水稻显性斑点叶突变体的鉴定和基因精细定位

    Institute of Scientific and Technical Information of China (English)

    郭丹; 施勇烽; 王惠梅; 张晓波; 宋莉欣; 徐霞; 贺彦; 郭梁; 吴建利

    2016-01-01

    A stable inherited rice spotted-leaf mutantHM113 was isolated from an EMS-induced IR64 mutant bank. Under natural conditions, brown lesions were observed on the leaves in three weeks after sowing and spread to the sheaths at the initial heading stage. Agronomic traits including the plant height, panicle length, number of panicles, number of filled grain/panicle, seed-setting rate and 1000-grain weight were decreased significantly inHM113. In addition, the photosynthetic pigment contents, net photo-synthetic rate and soluble protein content in the mutant were significantly lower than those in the wild type IR64, while the MDA content was similar to that in the wild-type. Activities of CAT and SOD were significantly lower and activity of POD was signifi-cantly higher in the mutant than in IR64. Histochemical analysis showed that cell death and ROS accumulation were occurred in and around the lesions inHM113. Furthermore, disease resistance to bacterial blight pathogens was significantly enhanced in the mutant in contrast to that in the wild type IR64. Expression of defense-related genes includingAOS2,PAL4,PR10,andPR1b was apparently up-regulated in the mutant. Genetic analysis indicated that the mutant trait was controlled by a novel single dominant nuclear gene, tentatively termedas SplHM113, which was detected to be located in a region around 308 kb flanked by RM21605 and RM418 on the long arm of chromosome 7. The data and populations obtained in the present study would facilitate the isolation and functional analysis ofSplHM113.%通过EMS (ethane methyl sulfonate)诱变籼稻品种IR64获得一个稳定遗传的显性斑点叶突变体HM113。在大田环境下,突变体褐色斑点在播种后3周的叶片上产生,始穗期扩散至叶鞘。与野生型IR64相比,突变体HM113的株高、结实率和千粒重等农艺性状显著下降,光合色素含量、净光合速率和可溶性蛋白含量显著降低。同时突

  20. Mutant DISC1 affects methamphetamine-induced sensitization and conditioned place preference: a comorbidity model.

    Science.gov (United States)

    Pogorelov, Vladimir M; Nomura, Jun; Kim, Jongho; Kannan, Geetha; Ayhan, Yavuz; Yang, Chunxia; Taniguchi, Yu; Abazyan, Bagrat; Valentine, Heather; Krasnova, Irina N; Kamiya, Atsushi; Cadet, Jean Lud; Wong, Dean F; Pletnikov, Mikhail V

    2012-03-01

    Genetic factors involved in neuroplasticity have been implicated in major psychiatric illnesses such as schizophrenia, depression, and substance abuse. Given its extended interactome, variants in the Disrupted-In-Schizophrenia-1 (DISC1) gene could contribute to drug addiction and psychiatric diseases. Thus, we evaluated how dominant-negative mutant DISC1 influenced the neurobehavioral and molecular effects of methamphetamine (METH). Control and mutant DISC1 mice were studied before or after treatment with non-toxic escalating dose (ED) of METH. In naïve mice, we assessed METH-induced conditioned place preference (CPP), dopamine (DA) D2 receptor density and the basal and METH-induced activity of DISC1 partners, AKT and GSK-3β in the ventral striatum. In ED-treated mice, 4 weeks after METH treatment, we evaluated fear conditioning, depression-like responses in forced swim test, and the basal and METH-induced activity of AKT and GSK-3β in the ventral striatum. We found impairment in METH-induced CPP, decreased DA D2 receptor density and altered METH-induced phosphorylation of AKT and GSK-3β in naïve DISC1 female mice. The ED regimen was not neurotoxic as evidenced by unaltered brain regional monoamine tissue content. Mutant DISC1 significantly delayed METH ED-produced sensitization and affected drug-induced phosphorylation of AKT and GSK-3β in female mice. Our results suggest that perturbations in DISC1 functions in the ventral striatum may impact the molecular mechanisms of reward and sensitization, contributing to comorbidity between drug abuse and major mental diseases.

  1. Salt Stress-induced Programmed Cell Death in Rice Root Tip Cells

    Institute of Scientific and Technical Information of China (English)

    Jian-You Li; Ai-Liang Jiang; Wei Zhang

    2007-01-01

    Salt stressed rice root tips were used to investigate the changes of reactive oxygen species (ROS) and antioxidant enzymes at the early stages of programmed cell death (PCD). The results indicated that 500 mmol/L NaCl treatment could lead to specific features of PCD in root tips, such as DNA ladder, nuclear condense and deformation, and transferase mediated dUTP nick end labeling positive reaction, which were initiated at 4 h of treatment and progressed thereafter. Cytochrome c release from mitochondria into cytoplasm was also observed, which occurred at 2 h and was earlier than the above nuclear events. In the very early phase of PCD, an immediate burst in hydrogen peroxide and superoxide anion production rate was accompanied by two-phase changes of superoxide dismutases and ascorbate peroxidase. A short period of increase in the activity was followed by prolonged impairment. Thus,we conclude that salt can induce PCD in rice root tip cells, and propose that in the early phase of rice root tip cell PCD, salt stress-induced oxidative burst increased the antioxidant enzyme activity, which, In turn, scavenged the ROS and abrogated PCD. Also, when the stress is prolonged, the antioxidant system is damaged and accumulated ROS induces the PCD process, which leads to cytochrome c release and nuclear change.

  2. Fe deficiency induced changes in rice (Oryza sativa L.) thylakoids.

    Science.gov (United States)

    Wang, Yuwen; Xu, Chao; Li, Kang; Cai, Xiaojie; Wu, Min; Chen, Guoxiang

    2017-01-01

    Iron deficiency is an important abiotic stress that limits productivity of crops all over the world. We selected a hybrid rice (Oryza sativa L.), LYPJ, which is super high-yield and widely cultured in China, to investigate changes in the components and structure of thylakoid membranes and photosynthetic performance in response to iron deficiency. Our results demonstrated that photosystem I (PSI) is the primary target for iron deficiency, while the changes in photosystem II (PSII) are important for rebuilding a balance in disrupted energy utilization and dissipation caused by differential degradation of photosynthetic components. The result of immunoblot analysis suggested that the core subunit PsaA declined drastically, while PsbA remained relatively stable. Furthermore, several organizational changes of the photosynthetic apparatus were found by BN-PAGE, including a marked decrease in the PSI core complexes, the Cytb 6 /f complex, and the trimeric form of the LHCII antenna, consistent with the observed unstacking grana. The fluorescence induction analysis indicated a descending PSII activity with energy dissipation enhanced markedly. In addition, we proposed that the crippled CO2 assimilation could be compensated by the enhanced of phosphoenolpyruvate carboxylase (PEPC), which is suggested by the decreased ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) and photosynthetic efficiency.

  3. Promoter of soybean early nodulin gene enod2B is induced by rhizobial Nod factors in transgenic rice

    Institute of Scientific and Technical Information of China (English)

    WANG Yanzhang; YU Guanqiao; SHEN Shanjiong(San Chiun Shen); ZHU Jiabi

    2004-01-01

    Nod factors, which are signaling molecules produced by Rhizobia, are the principal determinants of host specificity in Rhizobium-legume symbiosis. Nod factors can elicit a number of characteristic developmental responses in the roots of legumes, such as depolarization of the membrane potential in epidermal cells, specific expression of early nodulin genes and changes in the flux of calcium in root hairs, deformation of root hairs, cell division in the root cortex and formation of the nodule primordium. Whether the rice plant can respond to signaling molecules (I.e. Nod factors) is an important question, as it could establish the potential for symbiotic nitrogen fixation in rice. The promoter of the soybean (Glycine max) early nodulin gene Gmenod2B fused to the β-glucuronidase (GUS) reporter gene was used as a molecular marker to explore whether Nod factors can be recognized by rice cells as signaling molecules. Transgenic rice plants harboring the chimeric gene Gmenod2BP-GUS were obtained via an Agrobacterium tumefaciens-mediated system. NodNGR factors produced by a broad-host-range Rhizobium strain NGR234(pA28) were used as probes to investigate the activity of the Gmenod2B promoter in rice. Our results showed that the early nodulin gene Gmenod2B promoter was induced by NodNGR factors in transgenic rice, and that it was specifically expressed in rice plant roots. Moreover, GUS gene expression driven by the Gmenod2B promoter in transgenic rice was regulated by nitrogen status. These findings indicated that rice possessed the ability to respond to Nod factor signals, and that this signal transduction system resulted in activation of the Gmenod2B promoter. Thus, we predict that the Nod-factor inducible nodulin expression system, which is similar to Rhizobium-legume symbiosis, may also exist in rice.

  4. Metabolomic analysis of pathways related to rice grain chalkiness by a notched-belly mutant with high occurrence of white-belly grains.

    Science.gov (United States)

    Lin, Zhaomiao; Zhang, Xincheng; Wang, Zunxin; Jiang, Yutong; Liu, Zhenghui; Alexander, Danny; Li, Ganghua; Wang, Shaohua; Ding, Yanfeng

    2017-02-07

    Grain chalkiness is a highly undesirable trait deleterious to rice appearance and milling quality. The physiological and molecular foundation of chalkiness formation is still partially understood, because of the complex interactions between multiple genes and growing environments. We report the untargeted metabolomic analysis of grains from a notched-belly mutant (DY1102) with high percentage of white-belly, which predominantly occurs in the bottom part proximal to the embryo. Metabolites in developing grains were profiled on the composite platforms of UPLC/MS/MS and GC/MS. Sampling times were 5, 10, 15, and 20 days after anthesis, the critical time points for chalkiness formation. A total of 214 metabolites were identified, covering most of the central metabolic pathways and partial secondary pathways including amino acids, carbohydrates, lipids, cofactors, peptides, nucleotides, phytohormones, and secondary metabolites. A comparison of the bottom chalky part and the upper translucent part of developing grains of DY1102 resulted in 180 metabolites related to chalkiness formation. Generally, in comparison to the translucent upper part, the chalky endosperm had lower levels of metabolites regarding carbon and nitrogen metabolism for synthesis of storage starch and protein, which was accompanied by perturbation of pathways participating in scavenging of reactive oxygen species, osmorugulation, cell wall synthesis, and mineral ion homeostasis. Based on these results, metabolic mechanism of chalkiness formation is discussed, with the role of embryo highlighted.

  5. Induced defense responses in rice plants against small brown planthopper infestation

    Institute of Scientific and Technical Information of China (English)

    Canxing; Duan; Jiaojiao; Yu; Jianyu; Bai; Zhendong; Zhu; Xiaoming; Wang

    2014-01-01

    The small brown planthopper(SBPH), Laodelphax striatellus Fallén(Homoptera: Delphacidae), is a serious pest of rice(Oryza sativa L.) in China. To understand the mechanisms of rice resistance to SBPH, defense response genes and related defense enzymes were examined in resistant and susceptible rice varieties in response to SBPH infestation. The salicylic acid(SA) synthesis-related genes phenylalanine ammonia-lyase(PAL), NPR1, EDS1 and PAD4 were induced rapidly and to a much higher level in the resistant variety Kasalath than in the susceptible cultivar Wuyujing 3 in response to SBPH infestation. The expression level of PAL in the Kasalath rice at 12 h post-infestation(hpi) increased 7.52-fold compared with the un-infested control, and the expression level in Kasalath was 49.63, 87.18, 57.36 and 75.06 times greater than that in Wuyujing 3 at 24, 36, 48 and 72 hpi, respectively. However, the transcriptional levels of the jasmonic acid(JA) synthesis-related genes LOX and AOS2 in resistant Kasalath were significantly lower than in susceptible Wuyujing 3 at 24, 36, 48 and 72 hpi. The activities of the defense enzymes PAL, peroxidase(POD), and polyphenol oxidase(PPO) increased remarkably in Kasalath in response to SBPH infestation, and were closely correlated with the PAL gene transcript level. Our results indicated that the SA signaling pathway was activated in the resistant Kasalath rice variety in response to SBPH infestation and that the gene PAL played a considerable role in the resistance to SBPH.

  6. Induced defense responses in rice plants against small brown planthopper infestation

    Directory of Open Access Journals (Sweden)

    Canxing Duan

    2014-02-01

    Full Text Available The small brown planthopper (SBPH, Laodelphax striatellus Fallén (Homoptera: Delphacidae, is a serious pest of rice (Oryza sativa L. in China. To understand the mechanisms of rice resistance to SBPH, defense response genes and related defense enzymes were examined in resistant and susceptible rice varieties in response to SBPH infestation. The salicylic acid (SA synthesis-related genes phenylalanine ammonia-lyase (PAL, NPR1, EDS1 and PAD4 were induced rapidly and to a much higher level in the resistant variety Kasalath than in the susceptible cultivar Wuyujing 3 in response to SBPH infestation. The expression level of PAL in the Kasalath rice at 12 h post-infestation (hpi increased 7.52-fold compared with the un-infested control, and the expression level in Kasalath was 49.63, 87.18, 57.36 and 75.06 times greater than that in Wuyujing 3 at 24, 36, 48 and 72 hpi, respectively. However, the transcriptional levels of the jasmonic acid (JA synthesis-related genes LOX and AOS2 in resistant Kasalath were significantly lower than in susceptible Wuyujing 3 at 24, 36, 48 and 72 hpi. The activities of the defense enzymes PAL, peroxidase (POD, and polyphenol oxidase (PPO increased remarkably in Kasalath in response to SBPH infestation, and were closely correlated with the PAL gene transcript level. Our results indicated that the SA signaling pathway was activated in the resistant Kasalath rice variety in response to SBPH infestation and that the gene PAL played a considerable role in the resistance to SBPH.

  7. Assessment of a systematic expression profiling approach in ENU-induced mouse mutant lines.

    Science.gov (United States)

    Seltmann, Matthias; Horsch, Marion; Drobyshev, Alexei; Chen, Yali; de Angelis, Martin Hrabé; Beckers, Johannes

    2005-01-01

    Comparative genomewide expression profiling is a powerful tool in the effort to annotate the mouse genome with biological function. The systematic analysis of RNA expression data of mouse lines from the Munich ENU mutagenesis screen might support the understanding of the molecular biology of such mutants and provide new insights into mammalian gene function. In a direct comparison of DNA microarray experiments of individual versus pooled RNA samples of organs from ENU-induced mouse mutants, we provide evidence that individual RNA samples may outperform pools in some aspects. Genes with high biological variability in their expression levels (noisy genes) are identified as false positives in pooled samples. Evidence suggests that highly stringent housing conditions and standardized procedures for the isolation of organs significantly reduce biological variability in gene expression profiling experiments. Data on wild-type individuals demonstrate the positive effect of controlling variables such as social status, food intake before organ sampling, and stress with regard to reproducibility of gene expression patterns. Analyses of several organs from various ENU-induced mutant lines in general show low numbers of differentially expressed genes. We demonstrate the feasibility to detect transcriptionally affected organs employing RNA expression profiling as a tool for molecular phenotyping.

  8. Twist1 suppresses senescence programs and thereby accelerates and maintains mutant Kras-induced lung tumorigenesis.

    Directory of Open Access Journals (Sweden)

    Phuoc T Tran

    Full Text Available KRAS mutant lung cancers are generally refractory to chemotherapy as well targeted agents. To date, the identification of drugs to therapeutically inhibit K-RAS have been unsuccessful, suggesting that other approaches are required. We demonstrate in both a novel transgenic mutant Kras lung cancer mouse model and in human lung tumors that the inhibition of Twist1 restores a senescence program inducing the loss of a neoplastic phenotype. The Twist1 gene encodes for a transcription factor that is essential during embryogenesis. Twist1 has been suggested to play an important role during tumor progression. However, there is no in vivo evidence that Twist1 plays a role in autochthonous tumorigenesis. Through two novel transgenic mouse models, we show that Twist1 cooperates with Kras(G12D to markedly accelerate lung tumorigenesis by abrogating cellular senescence programs and promoting the progression from benign adenomas to adenocarcinomas. Moreover, the suppression of Twist1 to physiological levels is sufficient to cause Kras mutant lung tumors to undergo senescence and lose their neoplastic features. Finally, we analyzed more than 500 human tumors to demonstrate that TWIST1 is frequently overexpressed in primary human lung tumors. The suppression of TWIST1 in human lung cancer cells also induced cellular senescence. Hence, TWIST1 is a critical regulator of cellular senescence programs, and the suppression of TWIST1 in human tumors may be an effective example of pro-senescence therapy.

  9. Profilin 1 mutants form aggregates that induce accumulation of prion-like TDP-43.

    Science.gov (United States)

    Tanaka, Yoshinori; Hasegawa, Masato

    2016-07-03

    Mutations in the profilin 1 (PFN1) gene have been identified as a cause of familial amyotrophic lateral sclerosis (ALS), and neuropathological studies indicate that TDP-43 is accumulated in brains of patients with PFN1 mutation. Here, we investigated the role of PFN1 mutations in the formation of prion-like abnormal TDP-43. Expression of PFN1 with pathogenic mutations resulted in the formation of cytoplasmic aggregates positive for p62 and ubiquitin, and these aggregates sequestered endogenous TDP-43. TDP-43 accumulation was facilitated in the presence of proteasome or lysosome inhibitor. Co-expression of mutant PFN1 and TDP-43 increased the levels of detergent-insoluble and phosphorylated TDP-43, and this increase required the C-terminal region of TDP-43. Moreover, detergent-insoluble fractions prepared from cells expressing ALS-linked mutant PFN1 induced seed-dependent accumulation of TDP-43. These findings indicate that expression of PFN1 mutants induces accumulation of TDP-43, and promotes conversion of normal TDP-43 into an abnormal form. These results provide new insight into the mechanisms of TDP-43 proteinopathies and other diseases associated with amyloid-like protein deposition.

  10. Constitutive expression of pathogen-inducible OsWRKY31 enhances disease resistance and affects root growth and auxin response in transgenic rice plants

    Institute of Scientific and Technical Information of China (English)

    Juan Zhang; Youliang Peng; Zejian Guo

    2008-01-01

    WRKY transcription factors have many regulatory roles in response to biotic and abiotic stresses. In this study, we isolated a rice WRKY gene (OsWRKY31) that is induced by the rice blast fungus Magnaporthe grisea and auxin. This gene encodes a polypeptide of 211 amino-acid residues and belongs to a subgroup of the rice WRKY gene family that probably originated after the divergence of monocot and dicot plants. OsWRKY31 was found to be localized to the nucleus of onion epidermis cells to transiently express OsWRKY31-eGFP fusion protein. Analysis of 0sWRKY31 and its mutants fused with a Cal4 DNA-binding domain indicated that OsWRKY31 has transactivation activity in yeast. Overexpression of the OsWRKY31 gene was found to enhance resistance against infection with M. grisea, and the transgenic lines exhibited reduced lateral root formation and elongation compared with wild-type and RNAi plants. The lines with overexpression showed constitutive expression of many defense-related genes, such as PBZ1 and OsSci2, as well as early auxin-response genes, such as OsIAA4 and OsCrll genes. Furthermore, the plants with overexpression were less sensitive to exogenously supplied IBA, NAA and 2,4-D at high concentrations, suggesting that overexpression of the OsWRKY31 gene might alter the auxin response or transport. These results also suggest that OsWRKY31 might be a common component in the signal transduction pathways of the auxin response and the defense response in rice.

  11. Mineral phosphate solubilization by wild type and radiation induced mutants of pantoea dispersa and pantoea terrae

    Energy Technology Data Exchange (ETDEWEB)

    Murugesan, Senthilkumar; Lee, Young Keun; Kim, Jung Hun [Korea Atomic Energy Research Institte, Jeongeup (Korea, Republic of)

    2009-03-15

    Three mineral phosphate solubilizing (MPS) bacteria where isolated from rhizosphere soil samples of common bean and weed plants. 16S rDNA analysis indicated that the isolate P2 and P3 are closely related to Pantoea dispersa while isolate P4 is closely related to Pantoea terrae. Islates P2 and P3 recorded 381.60 {mu}g ml{sup -1} of tricalcium phosphate (TCP) solubilization respectively on 3 days incubation. Isolate P4 recorded the TCP solubilization of 215.85 {mu}g ml{sup -1} and the pH was dropped to 4.44 on 24 h incubation. Further incubation of P4 sharply decreased the available phosphorous to 28.94 {mu}g ml{sup -1} and pH level was raised to 6.32. Gamma radiation induced mutagenesis was carried out at LD{sub 99} dose of the wild type strains. The total of 14 mutant clones with enhanced MPS activity and 4 clones with decreased activity were selected based on solubilization index (SI) and phosphate solubilization assay. Mutant P2-M1 recorded the highest P-solubilizing potential among any other wild or mutnat clones by releasing 504.21 {mu}g ml{sup -1} of phosphorous i.e. 35% higher than its wild type by the end of day 5. A comparative evaluation of TCP solubilization by wild type isolates of Pantoea and their mutants, led to select three MPS mutant clones such as P2-M1, P3-M2 and P3-M4 with a potential to release >471.67 {mu}g ml{sup {sub {sup 1}}} of phosphorous from TCP. These over expressing mutant clones are considered as suitable candidates for biofertilization.

  12. Proinsulin misfolding and diabetes: mutant INS gene-induced diabetes of youth.

    Science.gov (United States)

    Liu, Ming; Hodish, Israel; Haataja, Leena; Lara-Lemus, Roberto; Rajpal, Gautam; Wright, Jordan; Arvan, Peter

    2010-11-01

    Type 1B diabetes (typically with early onset and without islet autoantibodies) has been described in patients bearing small coding sequence mutations in the INS gene. Not all mutations in the INS gene cause the autosomal dominant Mutant INS-gene Induced Diabetes of Youth (MIDY) syndrome, but most missense mutations affecting proinsulin folding produce MIDY. MIDY patients are heterozygotes, with the expressed mutant proinsulins exerting dominant-negative (toxic gain of function) behavior in pancreatic beta cells. Here we focus primarily on proinsulin folding in the endoplasmic reticulum, providing insight into perturbations of this folding pathway in MIDY. Accumulated evidence indicates that, in the molecular pathogenesis of the disease, misfolded proinsulin exerts dominant effects that initially inhibit insulin production, progressing to beta cell demise with diabetes.

  13. klf2ash317 Mutant Zebrafish Do Not Recapitulate Morpholino-Induced Vascular and Haematopoietic Phenotypes.

    Directory of Open Access Journals (Sweden)

    Peter Novodvorsky

    Full Text Available The zinc-finger transcription factor Krϋppel-like factor 2 (KLF2 transduces blood flow into molecular signals responsible for a wide range of responses within the vasculature. KLF2 maintains a healthy, quiescent endothelial phenotype. Previous studies report a range of phenotypes following morpholino antisense oligonucleotide-induced klf2a knockdown in zebrafish. Targeted genome editing is an increasingly applied method for functional assessment of candidate genes. We therefore generated a stable klf2a mutant zebrafish and characterised its cardiovascular and haematopoietic development.Using Transcription Activator-Like Effector Nucleases (TALEN we generated a klf2a mutant (klf2ash317 with a 14bp deletion leading to a premature stop codon in exon 2. Western blotting confirmed loss of wild type Klf2a protein and the presence of a truncated protein in klf2ash317 mutants. Homozygous klf2ash317 mutants exhibit no defects in vascular patterning, survive to adulthood and are fertile, without displaying previously described morphant phenotypes such as high-output cardiac failure, reduced haematopoetic stem cell (HSC development or impaired formation of the 5th accessory aortic arch. Homozygous klf2ash317 mutation did not reduce angiogenesis in zebrafish with homozygous mutations in von Hippel Lindau (vhl, a form of angiogenesis that is dependent on blood flow. We examined expression of three klf family members in wildtype and klf2ash317 zebrafish. We detected vascular expression of klf2b (but not klf4a or biklf/klf4b/klf17 in wildtypes but found no differences in expression that might account for the lack of phenotype in klf2ash317 mutants. klf2b morpholino knockdown did not affect heart rate or impair formation of the 5th accessory aortic arch in either wildtypes or klf2ash317 mutants.The klf2ash317 mutation produces a truncated Klf2a protein but, unlike morpholino induced klf2a knockdown, does not affect cardiovascular development.

  14. Heat-inducible Cre-lox system for marker excision in transgenic rice

    Indian Academy of Sciences (India)

    Abhilasha Khattri; Soumen Nandy; Vibha Srivastava

    2011-03-01

    The present study assessed the efficacy of a heat-inducible cre gene for conditional removal of the marker gene from a rice genome via Cre-lox recombination. A cre gene controlled by the soybean heat-shock promoter was introduced into the rice genome along with the recombination target (lox) construct. Cre-mediated recombination was expected to remove the marker gene and activate the promoter-less GUS gene. Six transgenic lines displayed well-regulated heat-inducible Cre activity in the callus. However, only one line that contained a single copy of the cre gene maintained this property in the regenerated plants and their progeny. Marker-free progeny were obtained from the plant that was heat-treated at the seedling stage, indicating the inheritance of the recombination `footprint’. The presence of the `footprint’ was verified by polymerase chain reaction and Southern analysis. Therefore, the cre gene controlled by the soybean heat-shock promoter is an effective tool for conditional removal of the marker gene in rice.

  15. Identification of a Herbicide Safener AD-67 Inducible cDNA in Rice

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    A herbicide safener AD-67 inducible cDNA was identified in an indica rice variety 9311 by mRNA differential display. The transcript was increased 6 h after sprayed with the safener solution, and 4 days later, the expression still could be detected. The fragment was recycled from the poly-gel and sequenced, and homologous analysis revealed the cDNA was 100% identical to some ESTs and cDNAs in rice database, and the amino acid sequence was 60-84% homologous to those of the Yippee genes in several eukaryotes. The fragment was extended to the whole long cDNA, and thus a primer pair was designed. RT-PCR analysis for the designed primer supported the induction result.

  16. Simulated space radiation-induced mutants in the mouse kidney display widespread genomic change.

    Science.gov (United States)

    Turker, Mitchell S; Grygoryev, Dmytro; Lasarev, Michael; Ohlrich, Anna; Rwatambuga, Furaha A; Johnson, Sorrel; Dan, Cristian; Eckelmann, Bradley; Hryciw, Gwen; Mao, Jian-Hua; Snijders, Antoine M; Gauny, Stacey; Kronenberg, Amy

    2017-01-01

    Exposure to a small number of high-energy heavy charged particles (HZE ions), as found in the deep space environment, could significantly affect astronaut health following prolonged periods of space travel if these ions induce mutations and related cancers. In this study, we used an in vivo mutagenesis assay to define the mutagenic effects of accelerated 56Fe ions (1 GeV/amu, 151 keV/μm) in the mouse kidney epithelium exposed to doses ranging from 0.25 to 2.0 Gy. These doses represent fluences ranging from 1 to 8 particle traversals per cell nucleus. The Aprt locus, located on chromosome 8, was used to select induced and spontaneous mutants. To fully define the mutagenic effects, we used multiple endpoints including mutant frequencies, mutation spectrum for chromosome 8, translocations involving chromosome 8, and mutations affecting non-selected chromosomes. The results demonstrate mutagenic effects that often affect multiple chromosomes for all Fe ion doses tested. For comparison with the most abundant sparsely ionizing particle found in space, we also examined the mutagenic effects of high-energy protons (1 GeV, 0.24 keV/μm) at 0.5 and 1.0 Gy. Similar doses of protons were not as mutagenic as Fe ions for many assays, though genomic effects were detected in Aprt mutants at these doses. Considered as a whole, the data demonstrate that Fe ions are highly mutagenic at the low doses and fluences of relevance to human spaceflight, and that cells with considerable genomic mutations are readily induced by these exposures and persist in the kidney epithelium. The level of genomic change produced by low fluence exposure to heavy ions is reminiscent of the extensive rearrangements seen in tumor genomes suggesting a potential initiation step in radiation carcinogenesis.

  17. Lanthanum Prevents Salt Stress-induced Programmed Cell Death in Rice Root Tip Cells by Controlling Early Induction Events

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    In a previous study, a salt stress-induced programmed cell death (PCD) model was established in rice root tip cells. Here,by using Wuyunjing 8th rice seedlings, the effects of lanthanum on salt stress-induced PCD early events were studied. The peroxidase (APX). Imidazole (20 mmol/L), the inhibitor of nicotinamide adenine dinucleotide phosphate-oxidase (NADPH oxidase), could alleviate the occurrence of PCD obviously, and such alleviation could be enhanced by the addition of La3+,indicating the involvement of NADPH oxidase in the salt stress-induced PCD process. Taken together, lanthanum could prevent salt stress-induced PCD occurrence in the rice root tip cells by blocking the calcium influx under stress, which was followed by inhibiting calcium-dependent NADPH oxidase activity to prevent O2·-production and, enhancing the cytosolic antioxidative enzyme activities to scavenge the reactive oxygen species.

  18. Excess copper induced oxidative stress and response of antioxidants in rice.

    Science.gov (United States)

    Thounaojam, Thorny Chanu; Panda, Piyalee; Panda, P; Mazumdar, Purabi; Mazumdar, P; Kumar, Devanand; Sharma, Gauri Dutta; Sharma, G D; Sahoo, Lingaraj; Sahoo, L; Panda, Sanjib Kumar; Panda, S K

    2012-04-01

    To investigate the effects of copper (Cu), rice plant (Oryza sativa. L. var. MSE-9) was treated with different Cu concentrations (0, 10, 50 and 100 μM) for 5 days in hydroponic condition. Gradual decrease in shoot and root growth was observed with the increase of Cu concentration and duration of treatment where maximum inhibition was recorded in root growth. Cu was readily absorbed by the plant though the maximum accumulation was found in root than shoot. Hydrogen peroxide (H(2)O(2)) production and lipid peroxidation were found increased with the elevated Cu concentration indicating excess Cu induced oxidative stress. Antioxidant enzymes superoxide dismutase (SOD), guaiacol peroxidase (GPX) and ascorbate peroxidase (APX) and glutathione reductase (GR) were effectively generated at the elevated concentrations of Cu though catalase (CAT) did not show significant variation with respect to control. Ascorbate (ASH), glutathione (GSH) and proline contents were also increased in all the Cu treated plants compared with the control. SOD isoenzyme was greatly affected by higher concentration of Cu and it was consistent with the changes of the activity assayed in solution. The present study confirmed that excess Cu inhibits growth, induced oxidative stress by inducing ROS formation while the stimulated antioxidative system appears adaptive response of rice plant against Cu induced oxidative stress. Moreover proline accumulation in Cu stress plant seems to provide additional defense against the oxidative stress.

  19. Rice From Mercury Contaminated Areas in Guizhou Province Induces c-jun Expression in Rat Brain

    Institute of Scientific and Technical Information of China (English)

    JIN-PING CHENG; WEN-HUA WANG; LI-YA QU; JIN-PING JIA; MIN ZHENG; XIU-LING JI; TAO YUAN

    2005-01-01

    Objective Mercury (Hg), as one of the priority pollutants and also a hot topic of frontier environmental research in many countries, has been paid higher attention in the world since the middle of the last century. Guizhou Province (at N24°30′-29°13′, E103°1′-109°30′, 1 100 m above the sea level, with subtropical humid climate) in southwest China is an important mercury production center. It has been found that the mercury content in most media of aquatics, soil, atmosphere and in biomass of corns, plants and animals, is higher than the national standard.The present study aims to explore the influence of mercury pollution on the health of local citizens. Methods The effect of rice from two mercury polluted experimental plots of Guizhou Province on the expression of c-jun mRNA in rat brain and c-jun protein in cortex, hippocampus and ependyma was observed using reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemical methods. Results The results showed that the mercury polluted rice induced expression of c-jun mRNA and its protein significantly. Selenium can reduce Hg uptake, an antagonism between selenium and mercury on the expression of c-jun mRNA and c-jun protein. Conclusion c-jun participates in the toxicity process of brain injury by mercury polluted rice, the expression of c- jun mRNA in brain, and c-jun protein in rat cortex and hippocampus can predict neurotoxicity of mercury polluted rice. People should be advised to be cautious in eating any kind of Hg-polluted foods. To reveal the relationship between c-jun induction and apoptosis, further examinations are required.

  20. Large genetic screens for gynogenesis and androgenesis haploid inducers in Arabidopsis thaliana failed to identify mutants

    Directory of Open Access Journals (Sweden)

    Virginie ePortemer

    2015-03-01

    Full Text Available Gynogenesis is a process in which the embryo genome originates exclusively from female origin, following embryogenesis stimulation by a male gamete. In contrast, androgenesis is the development of embryos that contain only the male nuclear genetic background. Both phenomena are of great interest in plant breeding as haploidisation is an efficient tool to reduce the length of breeding schemes to create varieties. Although few inducer lines have been described, the genetic control of these phenomena is poorly understood. We developed genetic screens to identify mutations that would induce gynogenesis or androgenesis in Arabidopsis thaliana. The ability of mutant pollen to induce either gynogenesis or androgenesis was tested by crossing mutagenized plants as males. Seedlings from these crosses were screened with recessive phenotypic markers, one genetically controlled by the female genome and another by the male genome. Positive and negative controls confirmed the unambiguous detection of both gynogenesis and androgenesis events. This strategy was applied to 1,666 EMS-mutagenised lines and 47 distant Arabidopsis strains. While an internal control suggested that the mutagenesis reached saturation, no gynogenesis or androgenesis inducer was found. However, spontaneous gynogenesis was observed at a frequency of 1/10,800. Altogether, these results suggest that no simple EMS-induced mutation in the male genome is able to induce gynogenesis or androgenesis in Arabidopsis.

  1. Abscisic acid-deficient sit tomato mutant responses to cadmium-induced stress.

    Science.gov (United States)

    Pompeu, Georgia B; Vilhena, Milca B; Gratão, Priscila L; Carvalho, Rogério F; Rossi, Mônica L; Martinelli, Adriana P; Azevedo, Ricardo A

    2017-03-01

    There is a very effective cross-talk between signals triggered by reactive oxygen species and hormonal responses in plants, activating proteins/enzymes likely to be involved in stress tolerance. Abscisic acid (ABA) is known as a stress hormone that takes part in the integration of signals. This work aimed to characterize the biochemical response and ultrastructural changes induced by cadmium (Cd) in the Micro-Tom (MT) sitiens ABA-deficient mutant (sit) and its wild-type (MT) counterpart. MT and sit plants were grown over a 96-h period in the presence of Cd (0, 10, and 100 μM CdCl2). The overall results indicated increases in lipid peroxidation, hydrogen peroxide content and in the activities of the key antioxidant enzymes such as catalase, glutathione reductase, and ascorbate peroxidase in both genotypes. On the other hand, no alteration was observed in chlorophyll content, while the activity of another antioxidant enzyme, superoxide dismutase, remained constant or even decreased in the presence of Cd. Roots and shoots of the sit mutant and MT were analyzed by light and transmission electron microscopy in order to characterize the structural changes caused by the exposure to this metal. Cd caused a decrease in intercellular spaces in shoots and a decrease in cell size in roots of both genotypes. In leaves, Cd affected organelle shape and internal organization of the thylakoid membranes, whereas noticeable increase in the number of mitochondria and vacuoles in MT and sit roots were observed. These results add new information that should help unravel the relative importance of ABA in regulating the cell responses to stressful conditions induced by Cd apart from providing the first characterization of this mutant to oxidative stress.

  2. Lignocellulolytic mutants of Pleurotus ostreatus induced by gamma-ray radiation and their genetic similarities

    Science.gov (United States)

    Lee, Y.-K.; Chang, H.-H.; Kim, J.-S.; Kim, J. K.; Lee, K.-S.

    2000-02-01

    To induce the lignocellulolytic mutants of Pleurotus ostreatus, the mycelia were irradiated by gamma-ray radiation to doses of 1-2 kGy. Five strains were isolated by the criteria of clamp connection, fruiting body formation, growth rate and activities of extracellular enzymes. All isolated strains were able to form the fruiting bodies and grew similarly to the control. The extracellular enzymes activities in liquid media of isolated strains were up to 10 times higher than the control. Genetic similarities of the isolated strains ranged from 64.4% to 93.3% of the control. From these results, it seems that the genetic diversity of P. ostreatus could be changed and useful strains be induced by gamma-ray radiation to recycle or reuse biowastes.

  3. Lignocellulolytic mutants of Pleurotus ostreatus induced by gamma-ray radiation and their genetic similarities

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Y-K. E-mail: yklee@nanum.kaeri.re.kr; Chang, H-H.; Kim, J-S.; Kim, J.K.; Lee, K-S

    2000-02-01

    To induce the lignocellulolytic mutants of Pleurotus ostreatus, the mycelia were irradiated by gamma-ray radiation to doses of 1-2 kGy. Five strains were isolated by the criteria of clamp connection, fruiting body formation, growth rate and activities of extracellular enzymes. All isolated strains were able to form the fruiting bodies and grew similarly to the control. The extracellular enzymes activities in liquid media of isolated strains were up to 10 times higher than the control. Genetic similarities of the isolated strains ranged from 64.4% to 93.3% of the control. From these results, it seems that the genetic diversity of P. ostreatus could be changed and useful strains be induced by gamma-ray radiation to recycle or reuse biowastes. (author)

  4. Anacardic acid induces apoptosis-like cell death in the rice blast fungus Magnaporthe oryzae.

    Science.gov (United States)

    Muzaffar, Suhail; Bose, Chinchu; Banerji, Ashok; Nair, Bipin G; Chattoo, Bharat B

    2016-01-01

    Anacardic acid (6-pentadecylsalicylic acid), extracted from cashew nut shell liquid, is a natural phenolic lipid well known for its strong antibacterial, antioxidant, and anticancer activities. Its effect has been well studied in bacterial and mammalian systems but remains largely unexplored in fungi. The present study identifies antifungal, cytotoxic, and antioxidant activities of anacardic acid in the rice blast fungus Magnaporthe oryzae. It was found that anacardic acid causes inhibition of conidial germination and mycelial growth in this ascomycetous fungus. Phosphatidylserine externalization, chromatin condensation, DNA degradation, and loss of mitochondrial membrane potential suggest that growth inhibition of fungus is mainly caused by apoptosis-like cell death. Broad-spectrum caspase inhibitor Z-VAD-FMK treatment indicated that anacardic acid induces caspase-independent apoptosis in M. oryzae. Expression of a predicted ortholog of apoptosis-inducing factor (AIF) was upregulated during the process of apoptosis, suggesting the possibility of mitochondria dependent apoptosis via activation of apoptosis-inducing factor. Anacardic acid treatment leads to decrease in reactive oxygen species rather than increase in reactive oxygen species (ROS) accumulation normally observed during apoptosis, confirming the antioxidant properties of anacardic acid as suggested by earlier reports. Our study also shows that anacardic acid renders the fungus highly sensitive to DNA damaging agents like ethyl methanesulfonate (EMS). Treatment of rice leaves with anacardic acid prevents M. oryzae from infecting the plant without affecting the leaf, suggesting that anacardic acid can be an effective antifungal agent.

  5. Studies on Biological Characterization of Rice Brittle Culm Mutants and Their Biomass Degradation Efficiency%特异水稻脆茎突变体生物学特性及生物质降解效率的研究

    Institute of Scientific and Technical Information of China (English)

    冯永清; 邹维华; 李丰成; 张晶; 张会; 谢国生; 涂媛苑; 路铁刚; 彭良才

    2013-01-01

      水稻脆茎突变体是一种重要的种质资源。为研究该突变体在能源作物培育中的利用潜力,比较分析了10个突变体及其野生型品种日本晴(NPB)在物理特性、细胞壁组成、农艺性状、生物质降解效率等方面的差异。结果表明:水稻脆茎突变体生长发育正常,但茎秆抗张力和株高低于 NPB;茎秆细胞壁纤维素含量降低,半纤维素含量增高,木质素含量增加不显著。细胞壁合成关键基因转录水平的变化与细胞壁组成一致。此外,水稻脆茎突变体的单株穗重与 NPB 差异不显著,有效穗数增加,生物质产量增加,抗倒伏性显著增强,秸秆易粉碎且在酸、碱预处理及纤维素复合酶酶解条件下的产糖量及纤维素降解效率显著高于 NPB。这些特异水稻脆茎突变体的表型优势揭示出其在水稻秸秆利用中的应用潜力,它们可以用于高产、生物质高效降解水稻品种的培育。%  Brittle culm mutants are considered as valuable germplasm resources for energy crop breeding in rice. This study analyzed biological characters and biomass digestibilities of 10 distinct rice brittle culm mutants. Compared with the wild type japonical variety NPB ( Nipponbare), all mutants displayed normal growth and development, and shorter plant height and lower tension stress. In terms of cell wall composition in the mature culm tissues, the mutants showed lower cellulose and higher hemicelluloses levels than that of the NPB and no significant increase is identified in lignin level, which were consistent with the RNA transcript levels of their major wall polymer synthases. Due to cell wall alternation, the rice mutants appeared to have a stronger resistance to lodging than NPB did. However, all mutants and NPB were determined with unchanged single plant spike weight and higher number of effective panicles, suggesting that the mutants were not much affected with their seed

  6. Asexual reproduction induces a rapid and permanent loss of sexual reproduction capacity in the rice fungal pathogen Magnaporthe oryzae: results of in vitro experimental evolution assays

    Directory of Open Access Journals (Sweden)

    Saleh Dounia

    2012-03-01

    Full Text Available Abstract Background Sexual reproduction is common in eukaryotic microorganisms, with few species reproducing exclusively asexually. However, in some organisms, such as fungi, asexual reproduction alternates with episodic sexual reproduction events. Fungi are thus appropriate organisms for studies of the reasons for the selection of sexuality or clonality and of the mechanisms underlying this selection. Magnaporthe oryzae, an Ascomycete causing blast disease on rice, reproduces mostly asexually in natura. Sexual reproduction is possible in vitro and requires (i two strains of opposite mating types including (ii at least one female-fertile strain (i.e. a strain able to produce perithecia, the female organs in which meiosis occurs. Female-fertile strains are found only in limited areas of Asia, in which evidence for contemporary recombination has recently been obtained. We induced the forced evolution of four Chinese female-fertile strains in vitro by the weekly transfer of asexual spores (conidia between Petri dishes. We aimed to determine whether female fertility was rapidly lost in the absence of sexual reproduction and whether this loss was controlled genetically or epigenetically. Results All the strains became female-sterile after 10 to 19 rounds of selection under asexual conditions. As no single-spore isolation was carried out, the observed decrease in the production of perithecia reflected the emergence and the invasion of female-sterile mutants. The female-sterile phenotype segregated in the offspring of crosses between female-sterile evolved strains and female-fertile wild-type strains. This segregation was maintained in the second generation in backcrosses. Female-sterile evolved strains were subjected to several stresses, but none induced the restoration of female fertility. This loss of fertility was therefore probably due to genetic rather than epigenetic mechanisms. In competition experiments, female-sterile mutants produced similar

  7. Bacillus oryzicola sp. nov., an Endophytic Bacterium Isolated from the Roots of Rice with Antimicrobial, Plant Growth Promoting, and Systemic Resistance Inducing Activities in Rice.

    Science.gov (United States)

    Chung, Eu Jin; Hossain, Mohammad Tofajjal; Khan, Ajmal; Kim, Kyung Hyun; Jeon, Che Ok; Chung, Young Ryun

    2015-06-01

    Biological control of major rice diseases has been attempted in several rice-growing countries in Asia during the last few decades and its application using antagonistic bacteria has proved to be somewhat successful for controlling various fungal diseases in field trials. Two novel endophytic Bacillus species, designated strains YC7007 and YC7010(T), with anti-microbial, plant growth-promoting, and systemic resistance-inducing activities were isolated from the roots of rice in paddy fields at Jinju, Korea, and their multifunctional activities were analyzed. Strain YC7007 inhibited mycelial growth of major rice fungal pathogens strongly in vitro. Bacterial blight and panicle blight caused by Xanthomonas oryzae pv. oryzae (KACC 10208) and Burkholderia glumae (KACC 44022), respectively, were also suppressed effectively by drenching a bacterial suspension (10(7) cfu/ml) of strain YC7007 on the rhizosphere of rice. Additionally, strain YC7007 promoted the growth of rice seedlings with higher germination rates and more tillers than the untreated control. The taxonomic position of the strains was also investigated. Phylogenetic analyses based on 16S rRNA gene sequences indicated that both strains belong to the genus Bacillus, with high similarity to the closely related strains, Bacillus siamensis KACC 15859(T) (99.67%), Bacillus methylotrophicus KACC 13105(T) (99.65%), Bacillus amyloliquefaciens subsp. plantarum KACC 17177(T) (99.60%), and Bacillus tequilensis KACC 15944(T) (99.45%). The DNA-DNA relatedness value between strain YC7010(T) and the most closely related strain, B. siamensis KACC 15859(T) was 50.4±3.5%, but it was 91.5±11.0% between two strains YC7007 and YC7010(T), indicating the same species. The major fatty acids of two strains were anteiso-C15:0 and iso C15:0. Both strains contained MK-7 as a major respiratory quinone system. The G+C contents of the genomic DNA of two strains were 50.5 mol% and 51.2 mol%, respectively. Based on these polyphasic studies

  8. Bacillus oryzicola sp. nov., an Endophytic Bacterium Isolated from the Roots of Rice with Antimicrobial, Plant Growth Promoting, and Systemic Resistance Inducing Activities in Rice

    Directory of Open Access Journals (Sweden)

    Eu Jin Chung

    2015-06-01

    Full Text Available Biological control of major rice diseases has been attempted in several rice-growing countries in Asia during the last few decades and its application using antagonistic bacteria has proved to be somewhat successful for controlling various fungal diseases in field trials. Two novel endophytic Bacillus species, designated strains YC7007 and YC7010T, with anti-microbial, plant growth-promoting, and systemic resistance-inducing activities were isolated from the roots of rice in paddy fields at Jinju, Korea, and their multifunctional activities were analyzed. Strain YC7007 inhibited mycelial growth of major rice fungal pathogens strongly in vitro. Bacterial blight and panicle blight caused by Xanthomonas oryzae pv. oryzae (KACC 10208 and Burkholderia glumae (KACC 44022, respectively, were also suppressed effectively by drenching a bacterial suspension (10⁷ cfu/ml of strain YC7007 on the rhizosphere of rice. Additionally, strain YC7007 promoted the growth of rice seedlings with higher germination rates and more tillers than the untreated control. The taxonomic position of the strains was also investigated. Phylogenetic analyses based on 16S rRNA gene sequences indicated that both strains belong to the genus Bacillus, with high similarity to the closely related strains, Bacillus siamensis KACC 15859T (99.67%, Bacillus methylotrophicus KACC 13105T (99.65%, Bacillus amyloliquefaciens subsp. plantarum KACC 17177T (99.60%, and Bacillus tequilensis KACC 15944T (99.45%. The DNA-DNA relatedness value between strain YC7010T and the most closely related strain, B. siamensis KACC 15859T was 50.4±3.5%, but it was 91.5±11.0% between two strains YC7007 and YC7010T, indicating the same species. The major fatty acids of two strains were anteiso-C15:0 and iso C15:0. Both strains contained MK-7 as a major respiratory quinone system. The G+C contents of the genomic DNA of two strains were 50.5 mol% and 51.2 mol%, respectively. Based on these polyphasic studies, the

  9. Mutability and importance of a hypermutable cell subpopulation that produces stress-induced mutants in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Caleb Gonzalez

    2008-10-01

    Full Text Available In bacterial, yeast, and human cells, stress-induced mutation mechanisms are induced in growth-limiting environments and produce non-adaptive and adaptive mutations. These mechanisms may accelerate evolution specifically when cells are maladapted to their environments, i.e., when they are are stressed. One mechanism of stress-induced mutagenesis in Escherichia coli occurs by error-prone DNA double-strand break (DSB repair. This mechanism was linked previously to a differentiated subpopulation of cells with a transiently elevated mutation rate, a hypermutable cell subpopulation (HMS. The HMS could be important, producing essentially all stress-induced mutants. Alternatively, the HMS was proposed to produce only a minority of stress-induced mutants, i.e., it was proposed to be peripheral. We characterize three aspects of the HMS. First, using improved mutation-detection methods, we estimate the number of mutations per genome of HMS-derived cells and find that it is compatible with fitness after the HMS state. This implies that these mutants are not necessarily an evolutionary dead end, and could contribute to adaptive evolution. Second, we show that stress-induced Lac(+ mutants, with and without evidence of descent from the HMS, have similar Lac(+ mutation sequences. This provides evidence that HMS-descended and most stress-induced mutants form via a common mechanism. Third, mutation-stimulating DSBs introduced via I-SceI endonuclease in vivo do not promote Lac(+ mutation independently of the HMS. This and the previous finding support the hypothesis that the HMS underlies most stress-induced mutants, not just a minority of them, i.e., it is important. We consider a model in which HMS differentiation is controlled by stress responses. Differentiation of an HMS potentially limits the risks of mutagenesis in cell clones.

  10. Genomic polymorphism and protein changes of soybean mutant induced by space environment

    Science.gov (United States)

    He, J.; Gao, Y.; Sun, Y.

    Soybean 194 4126 of excellent agricultural qualities such as high yield and rounder and wider leaf was selected in six generation after abroad recoverable satellite 15 days in 1996 from Soybean 72163 featured with long-leaf white-blossom grey-hair and infinitude-poding To explore the mechanisms of plant mutation induced by space environment we have experimented at genome and proteome level on Soybean 194 4126 and its control Soybean 72163 Amplified Fragment Length Polymorphism AFLP was used to identify mutated sits and the result shows that 36 polymorphic bands varying between 100 and 900 bp in 2022 DNA bands varying between 100 and 1500 bp have been amplified out of 64 pairs of primer combinations between mutant Soybean 194 4126 and the control plant So the mutation degree of DNA is 3 56 The protein two-dimensional electrophoresis 2-DE and peptide mass fingerprint PMF assays were used to investigate the difference of proteins in fruits and leaves between Soybean 194 4126 and its control Results indicate that 62 protein dots specially appear in Soybean 72163 and 39 dots specially in the mutant Soybean 194 4126 by image analysis software PDQuest in the 2-DE maps of soybean seeds Using PMF assay and protein data-base searching to investigate two distinct protein dots we found that the protein specially expressed in the seed of mutant Soybean 194 4126 may be Dehydrin and the other protein specially expressed in the seed of the control Soybean 72163 may be maturation-associated protein MAT1 Because Dehydrin and MAT1 are

  11. Increase of Fungal Pathogenicity and Role of Plant Glutamine in Nitrogen-Induced Susceptibility (NIS) To Rice Blast

    Science.gov (United States)

    Huang, Huichuan; Nguyen Thi Thu, Thuy; He, Xiahong; Gravot, Antoine; Bernillon, Stéphane; Ballini, Elsa; Morel, Jean-Benoit

    2017-01-01

    Highlight  Modifications in glutamine synthetase OsGS1-2 expression and fungal pathogenicity underlie nitrogen-induced susceptibility to rice blast. Understanding why nitrogen fertilization increase the impact of many plant diseases is of major importance. The interaction between Magnaporthe oryzae and rice was used as a model for analyzing the molecular mechanisms underlying Nitrogen-Induced Susceptibility (NIS). We show that our experimental system in which nitrogen supply strongly affects rice blast susceptibility only slightly affects plant growth. In order to get insights into the mechanisms of NIS, we conducted a dual RNA-seq experiment on rice infected tissues under two nitrogen fertilization regimes. On the one hand, we show that enhanced susceptibility was visible despite an over-induction of defense gene expression by infection under high nitrogen regime. On the other hand, the fungus expressed to high levels effectors and pathogenicity-related genes in plants under high nitrogen regime. We propose that in plants supplied with elevated nitrogen fertilization, the observed enhanced induction of plant defense is over-passed by an increase in the expression of the fungal pathogenicity program, thus leading to enhanced susceptibility. Moreover, some rice genes implicated in nitrogen recycling were highly induced during NIS. We further demonstrate that the OsGS1-2 glutamine synthetase gene enhances plant resistance to M. oryzae and abolishes NIS and pinpoint glutamine as a potential key nutrient during NIS. PMID:28293247

  12. Pseudomonas fluorescens WCS374r-induced systemic resistance in rice against Magnaporthe oryzae is based on pseudobactin-mediated priming for a salicylic acid-repressible multifaceted defense response.

    Science.gov (United States)

    De Vleesschauwer, David; Djavaheri, Mohammad; Bakker, Peter A H M; Höfte, Monica

    2008-12-01

    Selected strains of nonpathogenic rhizobacteria can reduce disease in foliar tissues through the induction of a defense state known as induced systemic resistance (ISR). Compared with the large body of information on ISR in dicotyledonous plants, little is known about the mechanisms underlying rhizobacteria-induced resistance in cereal crops. Here, we demonstrate the ability of Pseudomonas fluorescens WCS374r to trigger ISR in rice (Oryza sativa) against the leaf blast pathogen Magnaporthe oryzae. Using salicylic acid (SA)-nonaccumulating NahG rice, an ethylene-insensitive OsEIN2 antisense line, and the jasmonate-deficient mutant hebiba, we show that this WCS374r-induced resistance is regulated by an SA-independent but jasmonic acid/ethylene-modulated signal transduction pathway. Bacterial mutant analysis uncovered a pseudobactin-type siderophore as the crucial determinant responsible for ISR elicitation. Root application of WCS374r-derived pseudobactin (Psb374) primed naive leaves for accelerated expression of a pronounced multifaceted defense response, consisting of rapid recruitment of phenolic compounds at sites of pathogen entry, concerted expression of a diverse set of structural defenses, and a timely hyperinduction of hydrogen peroxide formation putatively driving cell wall fortification. Exogenous SA application alleviated this Psb374-modulated defense priming, while Psb374 pretreatment antagonized infection-induced transcription of SA-responsive PR genes, suggesting that the Psb374- and SA-modulated signaling pathways are mutually antagonistic. Interestingly, in sharp contrast to WCS374r-mediated ISR, chemical induction of blast resistance by the SA analog benzothiadiazole was independent of jasmonic acid/ethylene signaling and involved the potentiation of SA-responsive gene expression. Together, these results offer novel insights into the signaling circuitry governing induced resistance against M. oryzae and suggest that rice is endowed with multiple

  13. Photodynamic vaccination of hamsters with inducible suicidal mutants of Leishmania amazonensis elicits immunity against visceral leishmaniasis

    Science.gov (United States)

    Kumari, Shraddha; Samant, Mukesh; Khare, Prashant; Misra, Pragya; Dutta, Sujoy; Kolli, Bala Krishna; Sharma, Sharad; Chang, Kwang Poo; Dube, Anuradha

    2016-01-01

    Leishmania, naturally residing in the phagolysosomes of macrophages, is a suitable carrier for vaccine delivery. Genetic complementation of these trypanosomatid protozoa to partially rectify their defective heme-biosynthesis renders them inducible with δ-aminolevulinate to develop porphyria for selective photolysis, leaving infected host-cells unscathed. Delivery of released “vaccines” to antigen-presenting cells is thus expected to enhance immune response, while their self-destruction presents added advantages of safety. Such suicidal-L. amazonensis was found to confer immunoprophylaxis and immunotherapy on hamsters against L. donovani. Neither heat-killed nor live parasites without suicidal induction were effective. Photodynamic vaccination of hamsters with the suicidal-mutants reduced the parasite loads by 99% and suppressed the development of disease. These suppressions were accompanied by an increase in Leishmania-specific delayed-type hypersensitivity and lymphoproliferation as well as in the levels of splenic iNOS, IFN-γ and IL-12 expressions and of Leishmania-specific IgG2 in the serum. Moreover, a single intravenous administration of T-cells from vaccinated hamsters was shown to confer on naïve animals an effective cellular immunity against L. donovani challenges. The absence of lesion development at vaccination sites and parasites in the draining lymphnodes, spleen and liver further indicates that the suicidal mutants provide a safe platform for vaccine delivery against experimental visceral leishmaniasis. PMID:19053149

  14. Anomalies of autogamy in Paramecium tetraurelia temperature sensitive mutant induced by heat treatment.

    Science.gov (United States)

    Prajer, M; Iftode, F

    2001-01-01

    The run of autogamy in temperature sensitive mutant ts401, carrying also the mutation of trichocysts nd3a, at permissive (26 degrees C) and non-permissive temperature (35 degrees C) was studied and compared with this process in wild d4-2 stock of Paramecium tetraurelia. The effect of heat treatment on programmed nuclear and cortical events was investigated using cytological silver impregnation Fernandez-Galiano method and immunofluorescence technique with the application of two anti-alpha-tubulin antibodies. The appearance of cells with some large macronuclear fragments and an excess number of micronuclei suggests that nuclear differentiation is inhibited at restrictive temperature and that macronuclear regeneration takes place in this thermosensitive mutant. In such cells impaired oral reorganization was observed. After slight refeeding at the late autogamy, cells stopped at first postautogamous division were induced by heat treatment. The most striking feature of these abnormal divisions was the lack of a basal body duplication wave, which suggests that production of gamma-tubulin, structural protein being the essential component of the microtubule organizing centres, is disturbed at high temperature.

  15. Jasmonic acid carboxyl methyltransferase regulates development and herbivory-induced defense response in rice

    Institute of Scientific and Technical Information of China (English)

    Jinfeng Qi; Yonggen Lou; Jiancai Li; Xiu Han; Ran Li; Jianqiang Wu; Haixin Yu; Lingfei Hu; Yutao Xiao; Jing Lu

    2016-01-01

    Jasmonic acid (JA) and related metabolites play a key role in plant defense and growth. JA carboxyl methyltransferase (JMT) may be involved in plant defense and development by methylating JA to methyl jasmonate (MeJA) and thus influencing the concentrations of JA and related metabolites. However, no JMT gene has been well characterized in monocotyledon defense and development at the molecular level. After we cloned a rice JMT gene, OsJMT1, whose encoding protein was localized in the cytosol, we found that the recombinant OsJMT1 protein catalyzed JA to MeJA. OsJMT1 is up-regulated in response to infestation with the brown planthopper (BPH; Nilaparvata lugens). Plants in which OsJMT1 had been overexpressed (oe-JMT plants) showed reduced height and yield. These oe-JMT plants also exhibited increased MeJA levels but reduced levels of herbivore-induced JA and jasmonoyl-isoleucine (JA-Ile). The oe-JMT plants were more attractive to BPH female adults but showed increased resistance to BPH nymphs, probably owing to the different responses of BPH female adults and nymphs to the changes in levels of H2O2 and MeJA in oe-JMT plants. These results indicate that OsJMT1, by altering levels of JA and related metabolites, plays a role in regulating plant development and herbivore-induced defense responses in rice.

  16. Inhibition of photosynthesis and energy dissipation induced by water and high light stresses in rice.

    Science.gov (United States)

    Zhou, Yanhong; Lam, Hon Ming; Zhang, Jianhua

    2007-01-01

    Photoprotection mechanisms of rice plants were studied when its seedlings were subjected to the combined stress of water and high light. The imposition of water stress, induced by PEG 6000 which was applied to roots, resulted in substantial inhibition of stomatal conductance and net photosynthesis under all irradiance treatments. Under high light stress, the rapid decline of photosynthesis with the development of water stress was accompanied by decreases in the maximum velocity of RuBP carboxylation by Rubisco (V(cmax)), the capacity for ribulose-1,5-bisphosphate regeneration (J(max)), Rubisco and stromal FBPase activities, and the quantum efficiency of photosystem II, in the absence of any stomatal limitation of CO(2) supply. Water stress significantly reduced the energy flux via linear electron transport (J(PSII)), but increased light-dependent and DeltapH- and xanthophyll-mediated thermal dissipation (J(NPQ)). It is concluded that the drought-induced inhibition of photosynthesis under different irradiances in the rice was due to both diffusive and metabolic limitations. Metabolic limitation of photosynthesis may be related to the adverse effects of some metabolic processes and the oxidative damage to the chloroplast. Meanwhile, an enhanced thermal dissipation is an important process to minimize the adverse effects of drought and high irradiance when CO(2) assimilation is suppressed.

  17. Transposon Ac/Ds-induced chromosomal rearrangements at the rice OsRLG5 locus.

    Science.gov (United States)

    Xuan, Yuan Hu; Piao, Hai Long; Je, Byoung Il; Park, Soon Ju; Park, Su Hyun; Huang, Jin; Zhang, Jian Bo; Peterson, Thomas; Han, Chang-deok

    2011-12-01

    Previous studies have shown that pairs of closely-linked Ac/Ds transposable elements can induce various chromosomal rearrangements in plant genomes. To study chromosomal rearrangements in rice, we isolated a line (OsRLG5-161) that contains two inversely-oriented Ds insertions in OsRLG5 (Oryza sativa Receptor like kinase Gene 5). Among approximately 300 plants regenerated from OsRLG5-161 heterozygous seeds, 107 contained rearrangements including deletions, duplications and inversions of various sizes. Most rearrangements were induced by previously identified alternative transposition mechanism. Furthermore, we also detected a new class of rearrangements that contain juxtaposed inversions and deletions on the same chromosome. We propose that these novel alleles were generated by a previously unreported type of alternative transposition reactions involving the 5' and 3' termini of two inversely-oriented Ds elements located on the same chromatid. Finally, 11% of rearrangements contained inversions resulting from homologous recombination between the two inverted Ds elements in OsRLG5-161. The high frequency inheritance and great variety of rearrangements obtained suggests that the rice regeneration system results in a burst of transposition activity and a relaxation of the controls which normally limit the transposition competence of individual Ds termini. Together, these results demonstrate a greatly enlarged potential of the Ac/Ds system for plant chromosome engineering. © The Author(s) 2011. Published by Oxford University Press.

  18. Aerobic rice genotypes displayed greater adaptation to water-limited cultivation and tolerance to polyethyleneglycol-6000 induced stress.

    Science.gov (United States)

    Sandhu, Nitika; Jain, Sunita; Battan, K R; Jain, R K

    2012-01-01

    Water scarcity and drought have seriously threatened traditional rice cultivation practices in several parts of the world including India. In the present investigation, experiments were conducted to see if the water-efficient aerobic rice genotypes developed at UAS, Bangalore (MAS25, MAS26 and MAS109) and IRRI, Philippines (MASARB25 and MASARB868), are endowed with drought tolerance or not. A set of these aerobic and five lowland high-yielding (HKR47 and PAU201, Taraori Basmati, Pusa1121 and Pusa1460) indica rice genotypes were evaluated for: (i) yield and yield components under submerged and aerobic conditions in field, (ii) root morphology and biomass under aerobic conditions in pots in the nethouse, (iii) PEG-6000 (0, -1, -2 and -3 bar) induced drought stress at vegetative stage using a hydroponic culture system and (iv) polymorphism for three SSR markers associated with drought resistance traits. Under submerged conditions, the yield of aerobic rice genotypes declined by 13.4-20.1 % whereas under aerobic conditions the yield of lowland indica/Basmati rice varieties declined by 23-27 %. Under water-limited conditions in pots, aerobic rice genotypes had 54-73.8 % greater root length and 18-60 % higher fresh root biomass compared to lowland indica rice varieties. Notably, root length of MASARB25 was 35 % shorter than MAS25 whereas fresh and dry root biomass of MASARB25 was 10 % and 64 % greater than MAS25. The lowland indica were more sensitive to PEG-stress with a score of 5.9-7.6 for Basmati and 6.1-6.7 for non-aromatic indica rice varieties, than the aerobic rice genotypes (score 2.7-3.3). A set of three microsatellite DNA markers (RM212, RM302 and RM3825) located on chromosome 1 which has been shown to be associated with drought resistance was investigated in the present study. Two of these markers (RM212 and RM302) amplified a specific allele in all the aerobic rice genotypes which were absent in lowland indica rice genotypes.

  19. Progression of Hepatic Adenoma to Carcinoma in Ogg1 Mutant Mice Induced by Phenobarbital

    Directory of Open Access Journals (Sweden)

    Anna Kakehashi

    2017-01-01

    Full Text Available The carcinogenic potential of phenobarbital (PB was assessed in a mouse line carrying a mutant Mmh allele of the Mmh/Ogg1 gene encoding the enzyme oxoguanine DNA glycosylase (Ogg1 responsible for the repair of 8-hydroxy-2′-deoxyguanosine (8-OHdG. Mmh homozygous mutant (Ogg1−/− and wild-type (Ogg1+/+ male and female, 10-week-old, mice were treated with 500 ppm PB in diet for 78 weeks. Hepatocellular carcinomas (HCCs were found in PB-treated Ogg1−/− mice, while Ogg1+/+ animals developed only hepatocellular adenomas (HCAs at the same rate. This was coordinated with PB-induced significant elevation of 8-OHdG formation in DNA and cell proliferation in adjacent liver of Ogg1−/− mice. Proteome analysis predicted activation of transcriptional factor Nrf2 in the livers and HCAs of PB-administered Ogg1+/+ mice; however, its activation was insufficient or absent in the livers and HCCs of Ogg1−/− mice, respectively. Significant elevation of phase I and II metabolizing enzymes was demonstrated in both Ogg1−/− and Ogg1+/+ animals. Treatment of Ogg1−/− mice with PB resulted in significant elevation of cell proliferation in the liver. These results indicate that PB induced progression from HCA to HCC in Ogg1−/− mice, due to persistent accumulation of DNA oxidative base modifications and suppression of Nrf2-mediated oxidative stress response, resulting in significant elevation of cell proliferation.

  20. Mutant Brucella abortus membrane fusogenic protein induces protection against challenge infection in mice.

    Science.gov (United States)

    de Souza Filho, Job Alves; de Paulo Martins, Vicente; Campos, Priscila Carneiro; Alves-Silva, Juliana; Santos, Nathalia V; de Oliveira, Fernanda Souza; Menezes, Gustavo B; Azevedo, Vasco; Cravero, Silvio Lorenzo; Oliveira, Sergio Costa

    2015-04-01

    Brucella species can cause brucellosis, a zoonotic disease that causes serious livestock economic losses and represents a public health threat. The mechanism of virulence of Brucella spp. is not yet fully understood. Therefore, it is crucial to identify new molecules that serve as virulence factors to better understand this host-pathogen interplay. Here, we evaluated the role of the Brucella membrane fusogenic protein (Mfp) and outer membrane protein 19 (Omp19) in bacterial pathogenesis. In this study, we showed that B. abortus Δmfp::kan and Δomp19::kan deletion mutant strains have reduced persistence in vivo in C57BL/6 and interferon regulatory factor 1 (IRF-1) knockout (KO) mice. Additionally, 24 h after macrophage infection with a Δmfp::kan or Δomp19::kan strain expressing green fluorescent protein (GFP) approximately 80% or 65% of Brucella-containing vacuoles (BCVs) retained the late endosomal/lysosomal marker LAMP-1, respectively, whereas around 60% of BCVs containing wild-type S2308 were found in LAMP-1-negative compartments. B. abortus Δomp19::kan was attenuated in vivo but had a residual virulence in C57BL/6 and IRF-1 KO mice, whereas the Δmfp::kan strain had a lower virulence in these same mouse models. Furthermore, Δmfp::kan and Δomp19::kan strains were used as live vaccines. Challenge experiments revealed that in C57BL/6 and IRF-1 KO mice, the Δmfp::kan strain induced greater protection than the vaccine RB51 and protection similar that of vaccine S19. However, a Δomp19::kan strain induced protection similar to that of RB51. Thus, these results demonstrate that Brucella Mfp and Omp19 are critical for full bacterial virulence and that the Δmfp::kan mutant may serve as a potential vaccine candidate in future studies.

  1. MutMap-Gap: whole-genome resequencing of mutant F2 progeny bulk combined with de novo assembly of gap regions identifies the rice blast resistance gene Pii.

    Science.gov (United States)

    Takagi, Hiroki; Uemura, Aiko; Yaegashi, Hiroki; Tamiru, Muluneh; Abe, Akira; Mitsuoka, Chikako; Utsushi, Hiroe; Natsume, Satoshi; Kanzaki, Hiroyuki; Matsumura, Hideo; Saitoh, Hiromasa; Yoshida, Kentaro; Cano, Liliana M; Kamoun, Sophien; Terauchi, Ryohei

    2013-10-01

    Next-generation sequencing allows the identification of mutations responsible for mutant phenotypes by whole-genome resequencing and alignment to a reference genome. However, when the resequenced cultivar/line displays significant structural variation from the reference genome, mutations in the genome regions missing from the reference (gaps) cannot be identified by simple alignment. Here we report on a method called 'MutMap-Gap', which involves delineating a candidate region harboring a mutation of interest using the recently reported MutMap method, followed by de novo assembly, alignment, and identification of the mutation within genome gaps. We applied MutMap-Gap to isolate the blast resistant gene Pii from the rice cv Hitomebore using mutant lines that have lost Pii function. MutMap-Gap should prove useful for cloning genes that exhibit significant structural variations such as disease resistance genes of the nucleotide-binding site-leucine rich repeat (NBS-LRR) class.

  2. Gamma ray irradiation to roots of tea-plants and induced mutant system

    Energy Technology Data Exchange (ETDEWEB)

    Takeda, Yoshiyuki; Nekaku, Koji; Wada, Mitsumasa (National Research Inst. of Vegetables, Ornamental Plants and Tea, Ano, Mie (Japan))

    1990-11-01

    In order to utilize the useful mutation which is induced by irradiation for the breeding of tea-plants, the gamma-ray irradiation to the roots of tea-plants was carried out. The samples were the roots of tea-plants of four varieties dug up in February, 1984, and were adjusted to about 20 cm, then, put in the cold storage at 5degC for 9 months till the time of irradiation in November, 1984. However, a part of them was taken out in August, and planted in a field for 76 days to germinate, thereafter, used as the samples. The gamma-ray from a Co-60 source was irradiated in the radiation breeding laboratory of Agriculture Bioresources Research Institute at the total dose of 1, 2 and 3 kR and the dose rate of 500 R/h. The irradiated roots were planted as they are or in the state of being cut, and the rate of germination, the number of buds and the induced mutation were examined. Clear difference was not observed in the rate of germination and the number of buds between the irradiated samples and those without irradiation. The long roots were superior to the short roots regarding these items. The types of the induced mutation were mostly thin leaves, and also yellowing, mottling, fascination and so on occurred. The mutant system lacking trichomes on the back of new leaves is considered to be strong against tea anthracnose, and is valuable. (K.I.).

  3. [Repression of the enzyme inducible syntheses in Escherichia coli K12 mutant with a deleted ptsH gene].

    Science.gov (United States)

    Gershanovich, V N; Il'ina, T S; Rusina, O Iu; Iurovitskaia, N V; Bol'shakova, T N

    1977-01-01

    The genome of lambda phage with thermosensitive repressor was integrated into the pts region of the E. coli chromosome. Such a lysogenic culture behaves as a pts mutant at 30 degrees. Heating of cells of this strain leads to the induction of lambda prophage and formation of deletions in the pts region. A mutant with a deletion covering ptsH gene was isolated after prophage induction. The deletion nature of pts mutation was confirmed in genetic and biochemical experiments. It was shown that the deletion is small and does not involve ptsI and lig genes. The isolated deltaptsH mutant possesses all characteristics of pts mutants: pleiotropic impairment of transport and utilization of a number of carbohydrates, repression of the enzyme inducible synthesis and resistance to catabolite repression with glucose. These data (together with earlier ones) allow us to conclude that the phosphorylated form of HPr is involved (in direct of indirect manner/ in activation of DNA transcription.

  4. Differential effectiveness of Serratia plymuthica IC1270-induced systemic resistance against hemibiotrophic and necrotrophic leaf pathogens in rice

    Directory of Open Access Journals (Sweden)

    Höfte Monica M

    2009-01-01

    Full Text Available Abstract Background Induced resistance is a state of enhanced defensive capacity developed by a plant reacting to specific biotic or chemical stimuli. Over the years, several forms of induced resistance have been characterized, including systemic acquired resistance, which is induced upon localized infection by an avirulent necrotizing pathogen, and induced systemic resistance (ISR, which is elicited by selected strains of nonpathogenic rhizobacteria. However, contrary to the relative wealth of information on inducible defense responses in dicotyledoneous plants, our understanding of the molecular mechanisms underlying induced resistance phenomena in cereal crops is still in its infancy. Using a combined cytomolecular and pharmacological approach, we analyzed the host defense mechanisms associated with the establishment of ISR in rice by the rhizobacterium Serratia plymuthica IC1270. Results In a standardized soil-based assay, root treatment with IC1270 rendered foliar tissues more resistant to the hemibiotrophic pathogen Magnaporthe oryzae, causal agent of the devastating rice blast disease. Analysis of the cytological and biochemical alterations associated with restriction of fungal growth in IC1270-induced plants revealed that IC1270 primes rice for enhanced attacker-induced accumulation of reactive oxygen species (ROS and autofluorescent phenolic compounds in and near epidermal cells displaying dense cytoplasmic granulation. Similar, yet more abundant, phenotypes of hypersensitively dying cells in the vicinity of fungal hyphae were evident in a gene-for-gene interaction with an avirulent M. oryzae strain, suggesting that IC1270-inducible ISR and R protein conditioned effector-triggered immunity (ETI target similar defense mechanisms. Yet, this IC1270-inducible ISR response seems to act as a double-edged sword within the rice defense network as induced plants displayed an increased vulnerability to the necrotrophic pathogens Rhizoctonia

  5. Delayed degradation of chlorophylls and photosynthetic proteins in Arabidopsis autophagy mutants during stress-induced leaf yellowing.

    Science.gov (United States)

    Sakuraba, Yasuhito; Lee, Sang-Hwa; Kim, Ye-Sol; Park, Ohkmae K; Hörtensteiner, Stefan; Paek, Nam-Chon

    2014-07-01

    Plant autophagy, one of the essential proteolysis systems, balances proteome and nutrient levels in cells of the whole plant. Autophagy has been studied by analysing Arabidopsis thaliana autophagy-defective atg mutants, but the relationship between autophagy and chlorophyll (Chl) breakdown during stress-induced leaf yellowing remains unclear. During natural senescence or under abiotic-stress conditions, extensive cell death and early yellowing occurs in the leaves of atg mutants. A new finding is revealed that atg5 and atg7 mutants exhibit a functional stay-green phenotype under mild abiotic-stress conditions, but leaf yellowing proceeds normally in wild-type leaves under these conditions. Under mild salt stress, atg5 leaves retained high levels of Chls and all photosystem proteins and maintained a normal chloroplast structure. Furthermore, a double mutant of atg5 and non-functional stay-green nonyellowing1-1 (atg5 nye1-1) showed a much stronger stay-green phenotype than either single mutant. Taking these results together, it is proposed that autophagy functions in the non-selective catabolism of Chls and photosynthetic proteins during stress-induced leaf yellowing, in addition to the selective degradation of Chl-apoprotein complexes in the chloroplasts through the senescence-induced STAY-GREEN1/NYE1 and Chl catabolic enzymes.

  6. Cloning and functional validation of early inducible Magnaporthe oryzae responsive CYP76M7 promoter from rice

    Directory of Open Access Journals (Sweden)

    Joshitha eVijayan

    2015-05-01

    Full Text Available Cloning and functional characterization of plant pathogen inducible promoters is of great significance for their use in the effective management of plant diseases. The rice gene CYP76M7 was up regulated at 24, 48 and 72 hours post inoculation (hpi with two isolates of Magnaporthe oryzae Mo-ei-11 and Mo-ni-25. In this study, the promoter of CYP76M7 gene was cloned from rice cultivar HR-12, characterized and functionally validated. The Transcription Start Site of CYP76M7 was mapped at 45 bases upstream of the initiation codon. To functionally validate the promoter, 5′ deletion analysis of the promoter sequences was performed and the deletion fragments fused with the GUS reporter gene were used for generating stable transgenic Arabidopsis plants as well as for transient expression in rice. The spatial and temporal expression pattern of GUS in transgenic Arabidopsis plants and also in transiently expressed rice leaves revealed that the promoter of CYP76M7 gene was induced by M. oryzae. The induction of CYP76M7 promoter was observed at 24 hpi with M. oryzae. We report that, sequences spanning -222 bp to -520 bp, with the cluster of three W-boxes, two ASF1 motifs and a single GT-1 element may contribute to the M. oryzae inducible nature of CYP76M7 promoter. The promoter characterized in this study would be an ideal candidate for the overexpression of defence genes in rice for developing durable blast resistance rice lines.

  7. Rice hull smoke extract protects mice against a salmonella lipopolysaccharide-induced endotoxemia

    Science.gov (United States)

    Rice hulls accounting for 20% of the rice crop are a byproduct of post-harvest rice processing. Endotoxemia (sepsis, septic shock) is an inflammatory, virulent often fatal disease that results mainly from infection with Salmonella and other Gram-negative bacteria. The present study investigated the...

  8. Hypoxia-inducible genes encoding small EF-hand proteins in rice and tomato.

    Science.gov (United States)

    Otsuka, Chie; Minami, Ikuko; Oda, Kenji

    2010-01-01

    Rice has evolved metabolic and morphological adaptations to low-oxygen stress to grow in submerged paddy fields. To characterize the molecular components that mediate the response to hypoxia in rice, we identified low-oxygen stress early response genes by microarray analysis. Among the highly responsive genes, five genes, OsHREF1 to OsHREF5, shared strong homology. They encoded small proteins harboring two EF-hands, typical Ca(2+)-binding motifs. Homologous genes were found in many land plants, including SlHREF in tomato, which is also strongly induced by hypoxia. SlHREF induction was detected in both roots and shoots of tomato plants under hypoxia. With the exception of OsHREF5, OsHREF expression was unaffected by drought, salinity, cold, or osmotic stress. Fluorescent signals of green fluorescent protein-fused OsHREFs were detected in the cytosol and nucleus. Ruthenium red, an inhibitor of intracellular Ca(2+) release, repressed induction of OsHREF1-4 under hypoxia. The HREFs may be related to the Ca(2+) response to hypoxia.

  9. Rice bran oil prevents neuroleptic-induced extrapyramidal symptoms in rats: Possible antioxidant mechanisms

    Directory of Open Access Journals (Sweden)

    Noreen Samad

    2015-09-01

    Full Text Available Tardive dyskinesia (TD is one of the serious side effects of long-term antipsychotic treatment. Chronic treatment with neuroleptic leads to the development of abnormal oral movements called vacuous chewing movements (VCMs. The oxidative stress hypothesis of TD is one of the possible pathophysiologic models for TD. Preclinical and clinical studies of this hypothesis indicate that neurotoxic free radical production is likely to be a consequence of antipsychotic medication and is related to occurrence of TD. Oxidative stress is implicated in the pathophysiology of TD. Rats chronically treated with haloperidol orally at a dose of 0.2 mg/kg/day for a period of 5 weeks developed VCMs, which increased in a time-dependent manner as the treatment continued for 5 weeks. Motor coordination impairment started after the 1st week and was maximally impaired after 3 weeks and gradually returned to the 1st week value. Motor activity in an open field or home cage (activity box not altered. Administration of rice bran oil (antioxidant by oral tubes at a dose of 0.4 mL/day prevented the induction of haloperidol-elicited VCMs as well impairment of motor coordination. The results are discussed in the context of a protective role of antioxidant of rice bran oil in the prevention of haloperidol-induced extrapyramidal symptoms.

  10. Mutant human myocilin induces strain specific differences in ocular hypertension and optic nerve damage in mice.

    Science.gov (United States)

    McDowell, Colleen M; Luan, Tomi; Zhang, Zhang; Putliwala, Tasneem; Wordinger, Robert J; Millar, J Cameron; John, Simon W M; Pang, Iok-Hou; Clark, Abbot F

    2012-07-01

    Elevated intraocular pressure (IOP) is a causative risk factor for the development and progression of glaucoma. Glaucomatous mutations in myocilin (MYOC) damage the trabecular meshwork and elevate IOP in humans and in mice. Animal models of glaucoma are important to discover and better understand molecular pathogenic pathways and to test new glaucoma therapeutics. Although a number of different animal models of glaucoma have been developed and characterized, there are no true models of human primary open angle glaucoma (POAG). The overall goal of this work is to develop the first inducible mouse model of POAG using a human POAG relevant transgene (i.e. mutant MYOC) expression in mouse eyes to elevate IOP and cause pressure-induced damage to the optic nerve. Four mouse strains (A/J, BALB/cJ, C57BL/6J, and C3H/HeJ) were used in this study. Ad5.MYOC.Y437H (5 × 10(7) pfu) was injected intravitreally into one eye, with the uninjected contralateral eye serving as the control eye. Conscious IOP measurements were taken using a TonoLab rebound tonometer. Optic nerve damage was determined by scoring PPD stained optic nerve cross sections. Retinal ganglion cell and superior colliculus damage was assessed by Nissl stain cell counts. Intravitreal administration of viral vector Ad5.MYOC.Y437H caused a prolonged, reproducible, and statistically significant IOP elevation in BALB/cJ, A/J, and C57BL/6J mice. IOPs increased to approximately 25 mm Hg for 8 weeks (p death or superior colliculus damage at the 8-week time point in any of the strains tested. These results demonstrate strain dependent responses to Ad5.MYOC.Y437H-induced ocular hypertension and pressure-induced optic nerve damage. Copyright © 2012 Elsevier Ltd. All rights reserved.

  11. Anoxia-induced elevation of cytosolic Ca2+ concentration depends on different Ca2+ sources in rice and wheat protoplasts.

    Science.gov (United States)

    Yemelyanov, Vladislav V; Shishova, Maria F; Chirkova, Tamara V; Lindberg, Sylvia M

    2011-08-01

    The anoxia-dependent elevation of cytosolic Ca(2+) concentration, [Ca(2+)](cyt), was investigated in plants differing in tolerance to hypoxia. The [Ca(2+)](cyt) was measured by fluorescence microscopy in single protoplasts loaded with the calcium-fluoroprobe Fura 2-AM. Imposition of anoxia led to a fast (within 3 min) significant elevation of [Ca(2+)](cyt) in rice leaf protoplasts. A tenfold drop in the external Ca(2+) concentration (to 0.1 mM) resulted in considerable decrease of the [Ca(2+)](cyt) shift. Rice root protoplasts reacted upon anoxia with higher amplitude. Addition of plasma membrane (verapamil, La(3+) and EGTA) and intracellular membrane Ca(2+)-channel antagonists (Li(+), ruthenium red and cyclosporine A) reduced the anoxic Ca(2+)-accumulation in rice. Wheat protoplasts responded to anoxia by smaller changes of [Ca(2+)](cyt). In wheat leaf protoplasts, the amplitude of the Ca(2+)-shift little depended on the external level of Ca(2+). Wheat root protoplasts were characterized by a small shift of [Ca(2+)](cyt) under anoxia. Plasmalemma Ca(2+)-channel blockers had little effect on the elevation of cytosolic Ca(2+) in wheat protoplasts. Intact rice seedlings absorbed Ca(2+) from the external medium under anoxic treatment. On the contrary, wheat seedlings were characterized by leakage of Ca(2+). Verapamil abolished the Ca(2+) influx in rice roots and Ca(2+) efflux from wheat roots. Anoxia-induced [Ca(2+)](cyt) elevation was high particularly in rice, a hypoxia-tolerant species. In conclusion, both external and internal Ca(2+) stores are important for anoxic [Ca(2+)](cyt) elevation in rice, whereas the hypoxia-intolerant wheat does not require external sources for [Ca(2+)](cyt) rise. Leaf and root protoplasts similarly responded to anoxia, independent of their organ origin.

  12. A rice-based soluble form of a murine TNF-specific llama variable domain of heavy-chain antibody suppresses collagen-induced arthritis in mice.

    Science.gov (United States)

    Abe, Michiyo; Yuki, Yoshikazu; Kurokawa, Shiho; Mejima, Mio; Kuroda, Masaharu; Park, Eun Jeong; Scheller, Jürgen; Nakanishi, Ushio; Kiyono, Hiroshi

    2014-04-10

    Tumor necrosis factor alpha (TNF) plays a pivotal role in chronic inflammatory diseases such as rheumatoid arthritis and Crohn's disease. Although anti-TNF antibody therapy is now commonly used to treat patients suffering from these inflammatory conditions, the cost of treatment continues to be a concern. Here, we developed a rice transgenic system for the production of a llama variable domain of a heavy-chain antibody fragment (VHH) specific for mouse TNF in rice seeds (MucoRice-mTNF-VHH). MucoRice-mTNF-VHH was produced at high levels in the rice seeds when we used our most recent transgene-overexpression system with RNA interference technology that suppresses the production of major rice endogenous storage proteins while enhancing the expression of the transgene-derived protein. Production levels of mTNF-VHH in rice seeds reached an average of 1.45% (w/w). Further, approximately 91% of mTNF-VHH was released easily when the powder form of MucoRice-mTNF-VHH was mixed with PBS. mTNF-VHH purified by means of single-step gel filtration from rice PBS extract showed high neutralizing activity in an in vitro mTNF cytotoxicity assay using WEHI164 cells. In addition, purified mTNF-VHH suppressed progression of collagen-induced arthritis in mice. These results show that this rice-expression system is useful for the production of neutralizing VHH antibody specific for mTNF.

  13. Selection of Arabidopsis mutants overexpressing genes driven by the promoter of an auxin-inducible glutathione S-transferase gene.

    Science.gov (United States)

    van der Kop, D A; Schuyer, M; Pinas, J E; van der Zaal, B J; Hooykaas, P J

    1999-03-01

    Transgenic arabidopsis plants were isolated that contained a T-DNA construct in which the promoter of an auxin-inducible glutathione S-transferase (GST) gene from tobacco was fused to the kanamycin resistance (nptII) as well as to the beta-glucuronidase (gusA) reporter gene. Subsequently, seeds were treated with EMS to obtain mutants in which both reporter gene fusions were up-regulated. Northern analysis showed that the mRNA level of a related, endogenous auxin-inducible GST gene of Arabidopsis was increased in some of these mutants as well. Two of the gup (GST up-regulated) mutants were characterized in more detail and roughly mapped. Both had epinastic cotyledons and leaves, a phenotype that turned out to be linked to the gup mutation.

  14. Phenotypes induced by NM causing α-skeletal muscle actin mutants in fibroblasts, Sol 8 myoblasts and myotubes

    Directory of Open Access Journals (Sweden)

    Vandamme Drieke

    2009-03-01

    Full Text Available Abstract Background Nemaline myopathy is a neuromuscular disorder characterized by the presence of nemaline bodies in patient muscles. 20% of the cases are associated with α-skeletal muscle actin mutations. We previously showed that actin mutations can cause four different biochemical phenotypes and that expression of NM associated actin mutants in fibroblasts, myoblasts and myotubes induces a range of cellular defects. Findings We conducted the same biochemical experiments for twelve new actin mutants associated with nemaline myopathy. We observed folding and polymerization defects. Immunostainings of these and eight other mutants in transfected cells revealed typical cellular defects such as nemaline rods or aggregates, decreased incorporation in F-actin structures, membrane blebbing, the formation of thickened actin fibres and cell membrane blebbing in myotubes. Conclusion Our results confirm that NM associated α-actin mutations induce a range of defects at the biochemical level as well as in cultured fibroblasts and muscle cells.

  15. Proteomic analysis of beryllium-induced genotoxicity in an Escherichia coli mutant model system.

    Science.gov (United States)

    Taylor-McCabe, Kirsten J; Wang, Zaolin; Sauer, Nancy N; Marrone, Babetta L

    2006-03-01

    Beryllium is the second lightest metal, has a high melting point and high strength-to-weight ratio, and is chemically stable. These unique chemical characteristics make beryllium metal an ideal choice as a component material for a wide variety of applications in aerospace, defense, nuclear weapons, and industry. However, inhalation of beryllium dust or fumes induces significant health effects, including chronic beryllium disease and lung cancer. In this study, the mutagenicity of beryllium sulfate (BeSO(4)) and the comutagenicity of beryllium with a known mutagen 1-methyl-3-nitro-1-nitrosoguanidine (MNNG) were evaluated using a forward mutant detection system developed in Escherichia coli. In this system, BeSO(4) was shown to be weakly mutagenic alone and significantly enhanced the mutagenicity of MNNG up to 3.5-fold over MNNG alone. Based on these results a proteomic study was conducted to identify the proteins regulated by BeSO(4). Using the techniques of 2-DE and oMALDI-TOF MS, we successfully identified 32 proteins being differentially regulated by beryllium and/or MNNG in the E. coli test system. This is the first study to describe the proteins regulated by beryllium in vitro, and the results suggest several potential pathways for the focus of further research into the mechanisms underlying beryllium-induced genotoxicity.

  16. Applying genotyping (TILLING and phenotyping analyses to elucidate gene function in a chemically induced sorghum mutant population

    Directory of Open Access Journals (Sweden)

    Franks Cleve

    2008-10-01

    Full Text Available Abstract Background Sorghum [Sorghum bicolor (L. Moench] is ranked as the fifth most important grain crop and serves as a major food staple and fodder resource for much of the world, especially in arid and semi-arid regions. The recent surge in sorghum research is driven by its tolerance to drought/heat stresses and its strong potential as a bioenergy feedstock. Completion of the sorghum genome sequence has opened new avenues for sorghum functional genomics. However, the availability of genetic resources, specifically mutant lines, is limited. Chemical mutagenesis of sorghum germplasm, followed by screening for mutants altered in important agronomic traits, represents a rapid and effective means of addressing this limitation. Induced mutations in novel genes of interest can be efficiently assessed using the technique known as Targeting Induced Local Lesion IN Genomes (TILLING. Results A sorghum mutant population consisting of 1,600 lines was generated from the inbred line BTx623 by treatment with the chemical agent ethyl methanesulfonate (EMS. Numerous phenotypes with altered morphological and agronomic traits were observed from M2 and M3 lines in the field. A subset of 768 mutant lines was analyzed by TILLING using four target genes. A total of five mutations were identified resulting in a calculated mutation density of 1/526 kb. Two of the mutations identified by TILLING and verified by sequencing were detected in the gene encoding caffeic acid O-methyltransferase (COMT in two independent mutant lines. The two mutant lines segregated for the expected brown midrib (bmr phenotype, a trait associated with altered lignin content and increased digestibility. Conclusion TILLING as a reverse genetic approach has been successfully applied to sorghum. The diversity of the mutant phenotypes observed in the field, and the density of induced mutations calculated from TILLING indicate that this mutant population represents a useful resource for members of

  17. Morphological characterization and molecular mapping of an irradiation-induced Speckled mutant in the silkworm, Bombyx mori.

    Science.gov (United States)

    Tan, D; Tong, X-L; Hu, H; Wu, S-Y; Li, C-L; Xiong, G; Xiang, Z-H; Dai, F-Y; Lu, C

    2016-04-01

    Speckled (Spc), an X-ray-induced lethal mutant of Bombyx mori, exhibits a mosaic dark-brown-spotted larval epidermis in both sexes and egg-laying problems only in females. Here, we report the morphological characterization and molecular mapping of the Spc mutant. Morphological investigations revealed that the epidermal ultrastructure of the small, dark-brown spots was more dense than that of the white regions in both Spc/+ mutants and wild type, and that the lethality of the Spc/Spc mutants occurred during early embryogenesis. Furthermore, the ovarioles and ovipositor were disconnected in approximately 85.5% of Spc/+ females, a further 2.5% had a connection between the ovarioles and ovipositor that was too narrow to lay eggs. The remaining females showed a normal connection similar to that of the wild type. We successfully narrowed down the location of the Spc mutation to a region on chromosome 4 that was ∼1041 kb long. Gene-prediction analysis identified 25 candidate genes in this region. Chromosome structure analysis indicated that a ∼305 kb deletion was included in the mapping region. Temporal and spatial reverse transcription PCR (RT-PCR) analysis showed that several genes in the mapped region are associated with the Spc mutant. Although the genes responsible for the Spc mutation were not definitively identified, our results further the current understanding of the complex mechanism underlying the multiple morphological defects in Spc mutants.

  18. Inducible expression of a fusion gene encoding two proteinase inhibitors leads to insect and pathogen resistance in transgenic rice.

    Science.gov (United States)

    Quilis, Jordi; López-García, Belén; Meynard, Donaldo; Guiderdoni, Emmanuel; San Segundo, Blanca

    2014-04-01

    Plant proteinase inhibitors (PIs) are considered as candidates for increased insect resistance in transgenic plants. Insect adaptation to PI ingestion might, however, compromise the benefits received by transgenic expression of PIs. In this study, the maize proteinase inhibitor (MPI), an inhibitor of insect serine proteinases, and the potato carboxypeptidase inhibitor (PCI) were fused into a single open reading frame and introduced into rice plants. The two PIs were linked using either the processing site of the Bacillus thuringiensis Cry1B precursor protein or the 2A sequence from the foot-and-mouth disease virus (FMDV). Expression of each fusion gene was driven by the wound- and pathogen-inducible mpi promoter. The mpi-pci fusion gene was stably inherited for at least three generations with no penalty on plant phenotype. An important reduction in larval weight of Chilo suppressalis fed on mpi-pci rice, compared with larvae fed on wild-type plants, was observed. Expression of the mpi-pci fusion gene confers resistance to C. suppressalis (striped stem borer), one of the most important insect pest of rice. The mpi-pci expression systems described may represent a suitable strategy for insect pest control, better than strategies based on the use of single PI genes, by preventing insect adaptive responses. The rice plants expressing the mpi-pci fusion gene also showed enhanced resistance to infection by the fungus Magnaporthe oryzae, the causal agent of the rice blast disease. Our results illustrate the usefulness of the inducible expression of the mpi-pci fusion gene for dual resistance against insects and pathogens in rice plants.

  19. Evaluation on Inducible Effect of pOp6 Promoter in Transgenic Rice

    Institute of Scientific and Technical Information of China (English)

    Rouyi CHEN; Changxiang ZHENG; Jiang CHENG; Minna PAN; Mariena KETUDAT-CAIRNS

    2014-01-01

    In order to analyze the Os1bglu4 phenotype, the inducible promoter of the transgenic rice which knock-down the Os1bglu4 expression was assessed. The re-sult showed that 30 μM dexamethasone(DEX) had the stronger induction effect than 10 μM DEX by β-Glucuronidase (GUS) staining. qRT-PCR further verified the Os1bglu4 gene deletion. The effect of DEX and its solvent absolute ethanol on seed development was measured, and no significant effect was observed. The con-clusion is that final concentration of DEX at 30 μM is suitable for pOp6 promoter in-duction.

  20. Purification and Analysis of Abscisic Acid-Specifically-Inducible Proteins from Rice Callus

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Two ABA-specifically-inducible proteins from rice callus were isolated and purified by precipitation with 65-100 % saturated(NH4)2SO4, followed by the DEAE-sepharose, TSK-gel, and two-dimension electrophoresis. Iso-electric points (pl) of the proteins with the same molecular mass (24.5 kD) were 6.1 and 6.9, respectively. The Western blot analysis indicated that the proteins expressed in different tissues were obviously different. The A1 (pl 6.1) protein was only detected in calli treated with ABA and seed embryos (SE).However, the A2 (pl 6.9) protein was found not only in the calli treated with ABA and SE, but also in the white dry callus. Thus it suggested that the two proteins might play some important roles in the processes of seed embryo (or somatic embryo) formation.

  1. In vitro simulation studies of silica deposition induced by lignin from rice

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    To reveal the possible mechanism of silica deposition in higher plants, lignin was isolated from rice straw following a modified method to conduct a simulation experiment in vitro. UV and infrared absorption spectra showed that the substance had the unique characteristics of pure lignin. The presence of silicon in the precipitation was revealed by TEM (transmission electron microscopy) with EDXA (energy dispersive X-ray analysis) device. It was found that in the borax solution where lignin precipitation occurred silica-lignin co-precipitation was produced but not in the DMSO solution where lignin was broken into its composition compounds and did not precipitate. This means that it is macromolecular lignin itself but not its compounds that could induce silica deposition in higher plants.

  2. 水稻窄卷叶突变体 nrl(t)的遗传分析与基因定位%Genetic Analysis and Gene Mapping of a Narrow and Rolled Mutant nrl(t) in Rice

    Institute of Scientific and Technical Information of China (English)

    陈涛; 王才林; 刘燕清; 张亚东; 朱镇; 赵庆勇; 周丽慧; 姚姝; 于新; 赵凌

    2014-01-01

    为发掘、定位和克隆水稻窄卷叶相关突变基因,揭示叶片发育机理。对武运粳7号条纹叶枯病抗性改良品系中发现的窄卷叶突变体进行了表型观察、遗传分析及基因定位。结果表明,与野生型相比,突变体在叶片形态发生改变的同时,其株高、分蘖性、穗长、枝梗数、每穗实粒数、穗粒数、结实率、千粒质量等农艺性状也存在显著差异。遗传分析表明,该突变性状受一对隐性基因控制。以500株辐恢838/nrl(t)的F2隐性单株为定位群体,利用SSR标记将NRL(t)基因定位于水稻第11染色体短臂末端RM11-01、RM11-11之间,物理距离约为160 kb的范围内。通过水稻基因组注释数据库分析发现,在该区域存在26个预测的基因,测序分析表明已报道的homeobox3A基因可能是潜在的候选基因,这为进一步研究该基因奠定了基础。%Leaf morphology is one of key components for rice plant architecture , so excavation , mapping and cloning related mutant genes are very important for revealing mechanism of leaf development and breeding new rice varieties with ideotype and super-high yield.In this study,the narrow and rolled mutant ntl (t) was found from the im-proved line of rice stripe disease for Wuyunjing No .7 and the mutant phenotype genetic analysis and gene mapping was researched .Compared with wild type , the mutant showed multipe abnormal phenotypes , such as shorter leaf length and width,rolled leaf,dwarfism,more tillers,reduced panicle length,branch number,filled grain number per spike ,spikelets per spike , seed-setting rate and 1000-grain weight .Genetic analysis indicated that the mutant was controlled by a single recessive gene .Using 500 recessive mutant individuals in the F 2 population derived from the cross between Fuhui 838 and nrl(t) ,the gene NRL(t) was located to a confined region of 160 kb flanked by two mic-rosatellite markers RM11-01 and RM11-11 on the

  3. Arsenite treatment induces oxidative stress, upregulates antioxidant system, and causes phytochelatin synthesis in rice seedlings.

    Science.gov (United States)

    Mishra, Shruti; Jha, A B; Dubey, R S

    2011-07-01

    The effects of arsenite treatment on generation of reactive oxygen species, induction of oxidative stress, response of antioxidative system, and synthesis of phytochelatins were investigated in two indica rice (Oryza sativa L.) cvs. Malviya-36 and Pant-12 grown in sand cultures for a period of 5-20 days. Arsenite (As(2)O(3); 25 and 50 μM) treatment resulted in increased formation of superoxide anion (O (2) (.-) ), elevated levels of H(2)O(2) and thiobarbituric acid reactive substances, showing enhanced lipid peroxidation. An enhanced level of ascorbate (AA) and glutathione (GSH) was observed irrespective of the variation in the level of dehydroascorbate (DHA) and oxidized glutathione (GSSG) which in turn influenced redox ratios AA/DHA and GSH/GSSG. With progressive arsenite treatment, synthesis of total acid soluble thiols and phytochelatins (PC) increased in the seedlings. Among antioxidative enzymes, the activities of superoxide dismutase (EC 1.15.1.1), catalase (EC 1.11.1.6), total ascorbate peroxidase (APX, EC 1.11.1.11), chloroplastic ascorbate peroxidase, guaiacol peroxidase (EC 1.11.1.7), monodehydroascorbate reductase (EC 1.6.5.4), and glutathione reductase (EC 1.6.4.2) increased in arsenite treated seedlings, while dehyroascorbate reductase (EC 1.8.5.1) activity declined initially during 5-10 days and increased thereafter. Results suggest that arsenite treatment causes oxidative stress in rice seedlings, increases the levels of many enzymatic and non-enzymatic antioxidants, and induces synthesis of thiols and PCs, which may serve as important components in mitigating arsenite-induced oxidative damage.

  4. Comprehensive behavioral analysis of ENU-induced Disc1-Q31L and -L100P mutant mice

    Directory of Open Access Journals (Sweden)

    Shoji Hirotaka

    2012-02-01

    Full Text Available Abstract Background Disrupted-in-Schizophrenia 1 (DISC1 is considered to be a candidate susceptibility gene for psychiatric disorders, including schizophrenia, bipolar disorder, and major depression. A recent study reported that N-ethyl-N-nitrosourea (ENU-induced mutations in exon 2 of the mouse Disc1 gene, which resulted in the amino acid exchange of Q31L and L100P, caused an increase in depression-like behavior in 31 L mutant mice and schizophrenia-like behavior in 100P mutant mice; thus, these are potential animal models of psychiatric disorders. However, remaining heterozygous mutations that possibly occur in flanking genes other than Disc1 itself might induce behavioral abnormalities in the mutant mice. Here, to confirm the effects of Disc1-Q31L and Disc1-L100P mutations on behavioral phenotypes and to investigate the behaviors of the mutant mice in more detail, the mutant lines were backcrossed to C57BL/6JJcl through an additional two generations and the behaviors were analyzed using a comprehensive behavioral test battery. Results Contrary to expectations, 31 L mutant mice showed no significant behavioral differences when compared with wild-type control mice in any of the behavioral tests, including the Porsolt forced swim and tail suspension tests, commonly used tests for depression-like behavior. Also, 100P mutant mice exhibited no differences in almost all of the behavioral tests, including the prepulse inhibition test for measuring sensorimotor gating, which is known to be impaired in schizophrenia patients; however, 100P mutant mice showed higher locomotor activity compared with wild-type control mice in the light/dark transition test. Conclusions Although these results are partially consistent with the previous study in that there was hyperactivity in 100P mutant mice, the vast majority of the results are inconsistent with those of the previous study; this discrepancy may be explained by differences in the genetic background of the

  5. Comprehensive behavioral analysis of ENU-induced Disc1-Q31L and -L100P mutant mice.

    Science.gov (United States)

    Shoji, Hirotaka; Toyama, Keiko; Takamiya, Yoshihiro; Wakana, Shigeharu; Gondo, Yoichi; Miyakawa, Tsuyoshi

    2012-02-20

    Disrupted-in-Schizophrenia 1 (DISC1) is considered to be a candidate susceptibility gene for psychiatric disorders, including schizophrenia, bipolar disorder, and major depression. A recent study reported that N-ethyl-N-nitrosourea (ENU)-induced mutations in exon 2 of the mouse Disc1 gene, which resulted in the amino acid exchange of Q31L and L100P, caused an increase in depression-like behavior in 31 L mutant mice and schizophrenia-like behavior in 100P mutant mice; thus, these are potential animal models of psychiatric disorders. However, remaining heterozygous mutations that possibly occur in flanking genes other than Disc1 itself might induce behavioral abnormalities in the mutant mice. Here, to confirm the effects of Disc1-Q31L and Disc1-L100P mutations on behavioral phenotypes and to investigate the behaviors of the mutant mice in more detail, the mutant lines were backcrossed to C57BL/6JJcl through an additional two generations and the behaviors were analyzed using a comprehensive behavioral test battery. Contrary to expectations, 31 L mutant mice showed no significant behavioral differences when compared with wild-type control mice in any of the behavioral tests, including the Porsolt forced swim and tail suspension tests, commonly used tests for depression-like behavior. Also, 100P mutant mice exhibited no differences in almost all of the behavioral tests, including the prepulse inhibition test for measuring sensorimotor gating, which is known to be impaired in schizophrenia patients; however, 100P mutant mice showed higher locomotor activity compared with wild-type control mice in the light/dark transition test. Although these results are partially consistent with the previous study in that there was hyperactivity in 100P mutant mice, the vast majority of the results are inconsistent with those of the previous study; this discrepancy may be explained by differences in the genetic background of the mice, the laboratory environment, experimental protocols

  6. High Photosynthetic Efficiency of Leaf Colour Mutant of Rice (Oryza sativa L.)%水稻叶色突变体的高光合特性

    Institute of Scientific and Technical Information of China (English)

    欧立军

    2011-01-01

    标810S是温敏核不育系810S中的淡黄绿叶自然突变株,具有较高的光合速率.本文对突变体的光合色素、光合速率、荧光参数和光合关键酶活性等研究发现,标810S光合色素约为810S的一半,强光条件下标810S光合速率(Pn)比对照高,且没有明显的“午休”现象.标810S气孔导度(Gs)显著增加,叶绿素荧光动力学参数显示,标810S光量子转化效率高;光合酶1,5一二磷酸核酮糖羧化酶(RuBPCase)活性是对照的69.80%,磷酸烯醇式丙酮酸羧化酶(PEPCase)和NADP-苹果酸酶(NADH-ME)却比810S分别上升79.50%和69.06%.研究结果认为,光合色素含量下降是标810S叶片呈淡黄绿叶色的根本原因.突变体通过减少热耗散和提高光合电子传递速率来提高光能利用效率,为暗反应提供足够的同化力,而较高的PEPC活性和较大的气孔导度使其能更有效地固定C02,光反应和暗反应的协同使突变体具有较高的光合速率.%Biao 810S is a yellow-green leaf mutant of the thermosensitive genie male sterile (TGMS) rice which has higher photosynthetic rate. The photosynthetic characteristics of Biao 810S were studied with the wild type TGMS line 810S as a control to clarify the physiological basis of high photosynthetic efficiency and provide a theoretical basis for further utilization. The photosynthetic pigment, photosynthetic rate, fluorescence parameters and activity of photosynthetic key enzyme were measured. The content of photosynthetic pigment in Biao 810S was approximately half of that in 810S. However, the net photosynthetic rate of Biao 810S was higher than that of 810S under high intensity light and Biao 810S had no obvious "Midday depression" phenomenon. The stomatal conductance of Biao 810S was highly increased and the light quantum transformation efficiency was higher than that of 810S. The activity of ribulose-l,5-bisphosphate carboxylase (RuBPCase) in Biao 810S was 69.80% of that in 810S, but the

  7. Bacillomycin L and surfactin contribute synergistically to the phenotypic features of Bacillus subtilis 916 and the biocontrol of rice sheath blight induced by Rhizoctonia solani.

    Science.gov (United States)

    Luo, Chuping; Zhou, Huafei; Zou, Jincheng; Wang, Xiaoyu; Zhang, Rongsheng; Xiang, Yaping; Chen, Zhiyi

    2015-02-01

    The antagonistic activity of lipopeptides in Bacillus subtilis 916 has been well documented, yet relatively little is known about their mechanism in biofilm formation and environmental colonization. This study sought to examine the interaction of B. subtilis 916 on Rhizoctonia solani-infected rice sheath to elucidate the mechanism of colonization on plant leaves. Results showed that the mutants Δbac, Δsrf, and Δsrf + bac of B. subtilis 916, deficient in bacillomycin L and surfactin production, respectively, not only altered colony morphology but also changed swarming motility, reduced antagonistic activity, and decreased biofilm formation. In particular, biofilm formation in mutant Δbac, not Δsrf or Δsrf + bac, were restored with addition of surfactin and bacillomycin L at 10 and 50 μg/mL, respectively. Moreover, surfactin and bacillomycin L were able to restore or enhance swarming motility in the corresponding mutants at 10 μg/mL, respectively. With the aid of green fluorescent protein tagging, it was demonstrated that B. subtilis 916 formed a robust biofilm on the rice sheath blight lesion and colonized well on R. solani-infected rice sheath, while its corresponding mutants performed poorly. These observations also correlated with the rice cultivar pot experiments, in which B. subtilis 916 exhibited greater biocontrol than its mutants. Our results suggest that surfactin and bacillomycin L contribute differently but synergistically to the biocontrol of rice sheath blight in B. subtilis 916 through its antifungal activity, biofilm formation, and colonization.

  8. bfb, a novel ENU-induced blebs mutant resulting from a missense mutation in Fras1.

    Directory of Open Access Journals (Sweden)

    Kerry A Miller

    Full Text Available Fras1 is an extracellular matrix associated protein with essential roles in adhesion of epithelia and mesenchyme during early embryonic development. The adhesive function of Fras1 is achieved through interaction with a group of related proteins, Frem 1-3, and a cytoplasmic adaptor protein Grip1. Mutation of each of these proteins results in characteristic epithelial blistering and have therefore become known as "blebs" proteins. Human Fraser syndrome presents with a similar phenotype and the blebs mice have been instrumental in identification of the genetic basis of Fraser syndrome. We have identified a new ENU-induced blebs allele resulting from a novel missense mutation in Fras1. The resulting mouse strain, blood filled blisters (bfb, presents with a classic blebs phenotype but does not exhibit embryonic lethality typical of other blebs mutants and in addition, we report novel palate and sternal defects. Analysis of the bfb phenotype confirms the presence of epithelial-mesenchymal adhesion defects but also supports the emerging role of blebs proteins in regulating signalling during organogenesis. The bfb strain provides new opportunities to investigate the role of Fras1 in development.

  9. Transgenerational epimutations induced by multi-generation drought imposition mediate rice plant’s adaptation to drought condition

    Science.gov (United States)

    Zheng, Xiaoguo; Chen, Liang; Xia, Hui; Wei, Haibin; Lou, Qiaojun; Li, Mingshou; Li, Tiemei; Luo, Lijun

    2017-01-01

    Epigenetic mechanisms are crucial mediators of appropriate plant reactions to adverse environments, but their involvement in long-term adaptation is less clear. Here, we established two rice epimutation accumulation lines by applying drought conditions to 11 successive generations of two rice varieties. We took advantage of recent technical advances to examine the role of DNA methylation variations on rice adaptation to drought stress. We found that multi-generational drought improved the drought adaptability of offspring in upland fields. At single-base resolution, we discovered non-random appearance of drought-induced epimutations. Moreover, we found that a high proportion of drought-induced epimutations maintained their altered DNA methylation status in advanced generations. In addition, genes related to transgenerational epimutations directly participated in stress-responsive pathways. Analysis based on a cluster of drought-responsive genes revealed that their DNA methylation patterns were affected by multi-generational drought. These results suggested that epigenetic mechanisms play important roles in rice adaptations to upland growth conditions. Epigenetic variations have morphological, physiological and ecological consequences and are heritable across generations, suggesting that epigenetics can be considered an important regulatory mechanism in plant long-term adaptation and evolution under adverse environments. PMID:28051176

  10. Twist1 suppresses senescence programs and thereby accelerates and maintains mutant Kras-induced lung tumorigenesis

    DEFF Research Database (Denmark)

    Tran, Phuoc T; Shroff, Emelyn H; Burns, Timothy F

    2012-01-01

    KRAS mutant lung cancers are generally refractory to chemotherapy as well targeted agents. To date, the identification of drugs to therapeutically inhibit K-RAS have been unsuccessful, suggesting that other approaches are required. We demonstrate in both a novel transgenic mutant Kras lung cancer...

  11. Identification of rice (Oryza sativa L.) signal factors capable of inducing Agrobacterium vir gene expression

    Institute of Scientific and Technical Information of China (English)

    许东晖; 李宝健; 刘煜; 黄志纾; 古练权

    1996-01-01

    Two kinds of signal factors capable of inducing Agrobaorerium vir gene expression were purified and identified from leaf extracts of panicle-differentiating to flowering stage of rice (Oryza saliva L. cv. IR 72) detected by Agrobacterium vir(?) lacZ. fusion genes. The induction was similar to that observed with 5 μm actosyringone (AS). Based on the comprehensive analysis of the data by UV, IR, NMR, MS, HMQC and HMBC, the structures of these two signal factors are identified as 5, 7, 4’-trihydroxy-3’, 5’-dimethoxy-flavone (named tricin) and 5, 4’ -dihydroxy-3’, 5’ -dimethoxy-7- (β-D-glucosyloxy) -flavone, respectively. These results demonstrate that monocotyledonous plants do contain highly efficient vir gene inducing factors of Agrobacterium, and the reason why monocotyledonous plants are difficult to transform by Ayrobacterium is not due to absence of vir gene inducing factors, but due to the signal factors only produced in specific stage and tissue of monocotyledonous plants

  12. Assessing benzene-induced toxicity on wild type Euglena gracilis Z and its mutant strain SMZ.

    Science.gov (United States)

    Peng, Cheng; Arthur, Dionne M; Sichani, Homa Teimouri; Xia, Qing; Ng, Jack C

    2013-11-01

    Benzene is a representative member of volatile organic compounds and has been widely used as an industrial solvent. Groundwater contamination of benzene may pose risks to human health and ecosystems. Detection of benzene in the groundwater using chemical analysis is expensive and time consuming. In addition, biological responses to environmental exposures are uninformative using such analysis. Therefore, the aim of this study was to employ a microorganism, Euglena gracilis (E. gracilis) as a putative model to monitor the contamination of benzene in groundwater. To this end, we examined the wild type of E. gracilis Z and its mutant form, SMZ in their growth rate, morphology, chlorophyll content, formation of reactive oxygen species (ROS) and DNA damage in response to benzene exposure. The results showed that benzene inhibited cell growth in a dose response manner up to 48 h of exposure. SMZ showed a greater sensitivity compared to Z in response to benzene exposure. The difference was more evident at lower concentrations of benzene (0.005-5 μM) where growth inhibition occurred in SMZ but not in Z cells. We found that benzene induced morphological changes, formation of lipofuscin, and decreased chlorophyll content in Z strain in a dose response manner. No significant differences were found between the two strains in ROS formation and DNA damage by benzene at concentrations affecting cell growth. Based on these results, we conclude that E. gracilis cells were sensitive to benzene-induced toxicities for certain endpoints such as cell growth rate, morphological change, depletion of chlorophyll. Therefore, it is a potentially suitable model for monitoring the contamination of benzene and its effects in the groundwater.

  13. Huperzine A provides robust and sustained protection against induced seizures in Scn1a mutant mice

    Directory of Open Access Journals (Sweden)

    Jennifer C. Wong

    2016-10-01

    Full Text Available De novo loss-of-function mutations in the voltage-gated sodium channel (VGSC SCN1A (encoding Nav1.1 are the main cause of Dravet syndrome (DS, a catastrophic early-life encephalopathy associated with prolonged and recurrent early-life febrile seizures (FSs, refractory afebrile epilepsy, cognitive and behavioral deficits, and a 15-20% mortality rate. SCN1A mutations also lead to genetic epilepsy with febrile seizures plus (GEFS+, which is an inherited disorder characterized by early-life FSs and the development of a range of adult epilepsy subtypes. Current antiepileptic drugs often fail to protect against the severe seizures and behavioral and cognitive deficits found in patients with SCN1A mutations. To address the need for more efficacious treatments for SCN1A-derived epilepsies, we evaluated the therapeutic potential of Huperzine A, a naturally occurring reversible acetylcholinesterase inhibitor. In CF1 mice, Hup A (0.56 or 1 mg/kg was found to confer protection against 6 Hz-, pentylenetetrazole (PTZ-, and maximal electroshock (MES-induced seizures. Robust protection against 6 Hz-, MES-, and hyperthermia-induced seizures was also achieved following Hup A administration in mouse models of DS (Scn1a+/- and GEFS+ (Scn1aRH/+. Furthermore, Hup A-mediated seizure protection was sustained during 3 weeks of daily injections in Scn1aRH/+ mutants. Finally, we determined that the muscarinic and GABAA receptors play a role in Hup A-mediated seizure protection. These findings indicate that Hup A might provide a novel therapeutic strategy for increasing seizure resistance in DS and GEFS+, and more broadly, in other forms of refractory epilepsy.

  14. Abeta42 mutants with different aggregation profiles induce distinct pathologies in Drosophila.

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    Koichi Iijima

    Full Text Available Aggregation of the amyloid-beta-42 (Abeta42 peptide in the brain parenchyma is a pathological hallmark of Alzheimer's disease (AD, and the prevention of Abeta aggregation has been proposed as a therapeutic intervention in AD. However, recent reports indicate that Abeta can form several different prefibrillar and fibrillar aggregates and that each aggregate may confer different pathogenic effects, suggesting that manipulation of Abeta42 aggregation may not only quantitatively but also qualitatively modify brain pathology. Here, we compare the pathogenicity of human Abeta42 mutants with differing tendencies to aggregate. We examined the aggregation-prone, EOFAD-related Arctic mutation (Abeta42Arc and an artificial mutation (Abeta42art that is known to suppress aggregation and toxicity of Abeta42 in vitro. In the Drosophila brain, Abeta42Arc formed more oligomers and deposits than did wild type Abeta42, while Abeta42art formed fewer oligomers and deposits. The severity of locomotor dysfunction and premature death positively correlated with the aggregation tendencies of Abeta peptides. Surprisingly, however, Abeta42art caused earlier onset of memory defects than Abeta42. More remarkably, each Abeta induced qualitatively different pathologies. Abeta42Arc caused greater neuron loss than did Abeta42, while Abeta42art flies showed the strongest neurite degeneration. This pattern of degeneration coincides with the distribution of Thioflavin S-stained Abeta aggregates: Abeta42Arc formed large deposits in the cell body, Abeta42art accumulated preferentially in the neurites, while Abeta42 accumulated in both locations. Our results demonstrate that manipulation of the aggregation propensity of Abeta42 does not simply change the level of toxicity, but can also result in qualitative shifts in the pathology induced in vivo.

  15. Salt stress induced variation in DNA methylation pattern and its influence on gene expression in contrasting rice genotypes.

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    Ratna Karan

    Full Text Available BACKGROUND: Salinity is a major environmental factor limiting productivity of crop plants including rice in which wide range of natural variability exists. Although recent evidences implicate epigenetic mechanisms for modulating the gene expression in plants under environmental stresses, epigenetic changes and their functional consequences under salinity stress in rice are underexplored. DNA methylation is one of the epigenetic mechanisms regulating gene expression in plant's responses to environmental stresses. Better understanding of epigenetic regulation of plant growth and response to environmental stresses may create novel heritable variation for crop improvement. METHODOLOGY/PRINCIPAL FINDINGS: Methylation sensitive amplification polymorphism (MSAP technique was used to assess the effect of salt stress on extent and patterns of DNA methylation in four genotypes of rice differing in the degree of salinity tolerance. Overall, the amount of DNA methylation was more in shoot compared to root and the contribution of fully methylated loci was always more than hemi-methylated loci. Sequencing of ten randomly selected MSAP fragments indicated gene-body specific DNA methylation of retrotransposons, stress responsive genes, and chromatin modification genes, distributed on different rice chromosomes. Bisulphite sequencing and quantitative RT-PCR analysis of selected MSAP loci showed that cytosine methylation changes under salinity as well as gene expression varied with genotypes and tissue types irrespective of the level of salinity tolerance of rice genotypes. CONCLUSIONS/SIGNIFICANCE: The gene body methylation may have an important role in regulating gene expression in organ and genotype specific manner under salinity stress. Association between salt tolerance and methylation changes observed in some cases suggested that many methylation changes are not "directed". The natural genetic variation for salt tolerance observed in rice germplasm may be

  16. Immunogenicity and Cross-Protective Efficacy Induced by Outer Membrane Proteins from Salmonella Typhimurium Mutants with Truncated LPS in Mice

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    Qiong Liu

    2016-03-01

    Full Text Available Lipopolysaccharide (LPS is a major virulence factor present in the outer membrane of Salmonella enterica serovar Typhimurium (S. Typhimurium. Outer membrane proteins (OMPs from Salmonella show high immunogenicity and provide protection against Salmonella infection, and truncated LPS alters the outer membrane composition of the cell wall. In our previous study, we demonstrated that Salmonella mutants carrying truncated LPS failed to induce strong immune responses and cross-reaction to other enteric bacteria, due to their high attenuation and low colonization in the host. Therefore, we plan to investigate whether outer membrane proteins from Salmonella mutants with truncated LPS resulting from a series of nonpolar mutations, including ∆waaC12, ∆waaF15, ∆waaG42, ∆rfaH49, ∆waaI43, ∆waaJ44, ∆waaL46, ∆wbaP45 and ∆wzy-48, affect immunogenicity and provide protection against diverse Salmonella challenge. In this study, the immunogenicity and cross-protection efficiency of purified OMPs from all mutants were investigated to explore a potential OMP vaccine to protect against homologous or heterologous serotype Salmonella challenge. The results demonstrated that OMPs from three Salmonella mutants (∆waaC12, ∆waaJ44 and ∆waaL46 induced higher immune responses and provided good protection against homologous S. Typhimurium. The OMPs from these three mutants were also selected to determine the cross-protective efficacy against homologous and heterologous serotype Salmonella. Our results indicated that the mutant ∆waaC12 can elicit higher cross-reactivity and can provide good protection against S. Choleraesuis and S. Enteritidis infection and that the cross-reactivity may be ascribed to an antigen of approximately 18.4–30 kDa.

  17. Immunogenicity and Cross-Protective Efficacy Induced by Outer Membrane Proteins from Salmonella Typhimurium Mutants with Truncated LPS in Mice.

    Science.gov (United States)

    Liu, Qiong; Liu, Qing; Zhao, Xinxin; Liu, Tian; Yi, Jie; Liang, Kang; Kong, Qingke

    2016-03-22

    Lipopolysaccharide (LPS) is a major virulence factor present in the outer membrane of Salmonella enterica serovar Typhimurium (S. Typhimurium). Outer membrane proteins (OMPs) from Salmonella show high immunogenicity and provide protection against Salmonella infection, and truncated LPS alters the outer membrane composition of the cell wall. In our previous study, we demonstrated that Salmonella mutants carrying truncated LPS failed to induce strong immune responses and cross-reaction to other enteric bacteria, due to their high attenuation and low colonization in the host. Therefore, we plan to investigate whether outer membrane proteins from Salmonella mutants with truncated LPS resulting from a series of nonpolar mutations, including ∆waaC12, ∆waaF15, ∆waaG42, ∆rfaH49, ∆waaI43, ∆waaJ44, ∆waaL46, ∆wbaP45 and ∆wzy-48, affect immunogenicity and provide protection against diverse Salmonella challenge. In this study, the immunogenicity and cross-protection efficiency of purified OMPs from all mutants were investigated to explore a potential OMP vaccine to protect against homologous or heterologous serotype Salmonella challenge. The results demonstrated that OMPs from three Salmonella mutants (∆waaC12, ∆waaJ44 and ∆waaL46) induced higher immune responses and provided good protection against homologous S. Typhimurium. The OMPs from these three mutants were also selected to determine the cross-protective efficacy against homologous and heterologous serotype Salmonella. Our results indicated that the mutant ∆waaC12 can elicit higher cross-reactivity and can provide good protection against S. Choleraesuis and S. Enteritidis infection and that the cross-reactivity may be ascribed to an antigen of approximately 18.4-30 kDa.

  18. SCS118 Marques – New rice cultivar obtained through induced mutation

    Directory of Open Access Journals (Sweden)

    Moacir Antonio Schiocchet

    2014-03-01

    Full Text Available The new rice cultivar, SCS118 Marques, was obtained through gamma irradiation of SCSBRS Tio Taka cultivar. SCS118 Marques presents modern architecture, lodging resistance, late maturity cycle, moderate resistance to blast, high yield potential, long grains and very high cooking quality. Industrial tests performed with SCS118 Marques showed that grains are suitable for parboiling and white rice, and it is recommended to all rice-producing regions of Santa Catarina

  19. Molecular dissection of the roles of phytochrome in photoperiodic flowering in rice.

    Science.gov (United States)

    Osugi, Asami; Itoh, Hironori; Ikeda-Kawakatsu, Kyoko; Takano, Makoto; Izawa, Takeshi

    2011-11-01

    Phytochromes mediate the photoperiodic control of flowering in rice (Oryza sativa), a short-day plant. Recent molecular genetics studies have revealed a genetic network that enables the critical daylength response of florigen gene expression. Analyses using a rice phytochrome chromophore-deficient mutant, photoperiod sensitivity5, have so far revealed that within this network, phytochromes are required for expression of Grain number, plant height and heading date7 (Ghd7), a floral repressor gene in rice. There are three phytochrome genes in rice, but the roles of each phytochrome family member in daylength response have not previously been defined. Here, we revealed multiple action points for each phytochrome in the critical daylength response of florigen expression by using single and double phytochrome mutant lines of rice. Our results show that either phyA alone or a genetic combination of phyB and phyC can induce Ghd7 mRNA, whereas phyB alone causes some reduction in levels of Ghd7 mRNA. Moreover, phyB and phyA can affect Ghd7 activity and Early heading date1 (a floral inducer) activity in the network, respectively. Therefore, each phytochrome gene of rice has distinct roles, and all of the phytochrome actions coordinately control the critical daylength response of florigen expression in rice.

  20. Glycoprotein processing in mutants of HSV-1 that induce cell fusion

    Energy Technology Data Exchange (ETDEWEB)

    Person, S.; Kousoulas, K.G.; Knowles, R.W.; Read, G.S.; Holland, T.C.; Keller, P.M.; Warner, S.C.

    1982-01-01

    The synthesis of viral-specified glycoproteins, and their appearance on cell surfaces, were compared for cells infected either with syncytial mutants of HSV-1 or with the parental strains from which the mutants were derived. The mutants MP and tsB5, representatives of two different viral genes that affect fusion, were employed in these studies. Cells infected with either mutant gave rise to reduce surface labeling of major viral-specified glycoproteins throughout infection relative to the parental strains. Putative glycoprotein precursors were synthesized in similar amounts throughout infection in mutant and wild-type infected cells. However, during a chase period after a pulse of labeling, fully glycosylated species of glycoproteins accumulated more slowly in mutant than in parent infected cells. The effect was clearly visible early in MP-infected cells and by 9 hr after infection for tsB5-infected cells, and increased thereafter. Apparently the decrease in appearance of glycoproteins on the cell surface of mutant-infected cells is due to a decrease in their rate of processing. 26 references, 7 figures.

  1. Mutation breedings in ornamental plants. Technique used for radiation induced mutant in begonia, chrysanthemum, aberia and winter daphne

    Energy Technology Data Exchange (ETDEWEB)

    Matsubara, Hisao

    1984-03-01

    Several methods of obtaining somatic mutant plants by el-ray irradiation on pieces of tissues as in vitro adventitious bud technique or small cutting methods with repeated pruning are described. The irradiation to the adventitious buds in the small pieces of organ cultured in vitro and to the small cuttings are employed. Culture beds of agar or of Japanese Kanuma soil were used in vitro culture. In these experiments, Japanese Kanuma soil bed in in vitro culture worked well for root development and transplant of the induced mutants. Combination with in vitro culture and repeated pruning technique were used for isolation and fixation of solid somatic mutant from small sectorial mutation induced by irradiation. This method was successful for begonia, chrysanthemum, aberia and winter daphne. These data indicates that most of the induced mutant plants were non-chimeric, while a few others were chimeric. Among the new varieties, ''Gin-Sei'', ''Ryoku-Ha'', ''Big-Cross'', ''Kaede-Iron'', ''Mei-Fu-Hana-Tsukubane-Utsugi'' and ''Daphne-el-3'' are non-chimeric, and ''Mini-Mini-Iron'' and ''Orange-Iron'' are chimeric. Moreover, these new varieties have remarkably differed in size and in color pattern from original variety. From the experimental results of somatic mutation, it is indicated that plant tissue culture have enormous potential in radiation breeding and in rapid propagation of the somatic mutant. (author).

  2. DCP-LA neutralizes mutant amyloid beta peptide-induced impairment of long-term potentiation and spatial learning.

    Science.gov (United States)

    Nagata, Tetsu; Tomiyama, Takami; Tominaga, Takemi; Mori, Hiroshi; Yaguchi, Takahiro; Nishizaki, Tomoyuki

    2010-01-01

    Long-term potentiation (LTP) was monitored from the CA1 region of the intact rat hippocampus by delivering high frequency stimulation (HFS) to the Schaffer collateral commissural pathway. Intraventricular injection with mutant amyloid beta(1-42) peptide lacking glutamate-22 (Abeta(1-42)E22Delta), favoring oligomerization, 10 min prior to HFS, inhibited expression of LTP, with the potency more than wild-type amyloid beta(1-42) peptide. Intraperitoneal injection with the linoleic acid derivative 8-[2-(2-pentyl-cyclopropylmethyl)-cyclopropyl]-octanoic acid (DCP-LA) 70 min prior to HFS neutralized mutant Abeta(1-42)E22Delta peptide-induced LTP inhibition. In the water maze test, continuous intraventricular injection with mutant Abeta(1-42)E22Delta peptide for 14 days prolonged the acquisition latency as compared with that for control, with the potency similar to wild-type Abeta(1-42) peptide, and intraperitoneal injection with DCP-LA shortened the prolonged latency to control levels. The results of the present study indicate that DCP-LA neutralizes mutant Abeta(1-42)E22Delta peptide-induced impairment of LTP and spatial learning.

  3. Gamma-rays and EMS induced pentaphyllous mutant in black gram (Vigna mungo).

    Science.gov (United States)

    Singh, R K; Raghuvanshi, S S; Prakash, D

    1988-01-01

    Pentaphyllous mutants in black gram were isolated in M2 generation of a segregating family, irradiated at 20 kR. The genetic nature of mutants was tested by hybridizing with controls, and chi-square tests applied to the F2 population, proved it to be a monogenic recessive. The pentaphyllous mutant had a greater number of pods and leaves per plant and larger and more root nodules. It also showed improved nutritional value with increased seed protein percentage and no increase in TIA (trypsin inhibitor activity).

  4. Mildew-resistant mutants induced in North American two- and six-rowed malting barley cultivars

    DEFF Research Database (Denmark)

    Molina-Cano, J.L.; Simiand, J.P.; Sopena, A.;

    2003-01-01

    and were shown to have two new alleles at the mlo locus; these were designated, respectively, mlo31 and mlo32. Mutant URS2, showing partial resistance, was inherited as a dominant gene, but was not an allele at the Mla locus. The mean yield of each mutant was higher than that of its parental line......, but both URS1 and URS2 showed lower malt extract than Ursula. This lower extract might be due to the smaller grain size of the mutants that could, in turn, result from necrotic lesions in the leaves, as implied by the effects on grain yield....

  5. Toll-like receptor 4 mutant and null mice retain morphine-induced tolerance, hyperalgesia, and physical dependence.

    Directory of Open Access Journals (Sweden)

    Theresa Alexandra Mattioli

    Full Text Available The innate immune system modulates opioid-induced effects within the central nervous system and one target that has received considerable attention is the toll-like receptor 4 (TLR4. Here, we examined the contribution of TLR4 in the development of morphine tolerance, hyperalgesia, and physical dependence in two inbred mouse strains: C3H/HeJ mice which have a dominant negative point mutation in the Tlr4 gene rendering the receptor non-functional, and B10ScNJ mice which are TLR4 null mutants. We found that neither acute antinociceptive response to a single dose of morphine, nor the development of analgesic tolerance to repeated morphine treatment, was affected by TLR4 genotype. Likewise, opioid induced hyperalgesia and opioid physical dependence (assessed by naloxone precipitated withdrawal were not altered in TLR4 mutant or null mice. We also examined the behavioural consequence of two stereoisomers of naloxone: (- naloxone, an opioid receptor antagonist, and (+ naloxone, a purported antagonist of TLR4. Both stereoisomers of naloxone suppressed opioid induced hyperalgesia in wild-type control, TLR4 mutant, and TLR4 null mice. Collectively, our data suggest that TLR4 is not required for opioid-induced analgesic tolerance, hyperalgesia, or physical dependence.

  6. Laxative effects of fermented rice extract in rats with loperamide-induced constipation.

    Science.gov (United States)

    Choi, Jae-Suk; Kim, Joo Wan; Cho, Hyung-Rae; Kim, Ki-Young; Lee, Jong-Kwang; Sohn, Jae Hak; Ku, Sae-Kwang

    2014-12-01

    Constipation is a common problem in males and females. The aim of the present study was to evaluate the laxative effects of fermented rice extract (FRe) on rats with loperamide-induced constipation. FRe (100, 200 and 300 mg/kg) was administered orally once per day for six days following 1 h loperamide treatment. The laxative effects of FRe were compared with those of sodium picosulfate (S. picosulfate). Following the induction of constipation in the rats, a marked decrease was observed in the fecal pellet number and water content discharged over 24 h, the surface mucus thickness in the colonic lumen, intestinal charcoal transit ratio, thickness of the colonic mucosa and the number of mucus-producing cells, while an increase was observed in the number of fecal pellets remaining in the colonic lumen and their mean diameter, as compared with the normal vehicle control rats. These conditions were significantly alleviated following the administration of the three doses of FRe when compared with the loperamide control group. However, the alleviating effects were lower than those of S. picosulfate, with the exception of the intestinal charcoal transit ratio. Similar effects on the intestinal charcoal transit ratio were detected for the three doses of FRe when compared with the S. picosulfate-treated rats. In conclusion, the results indicated that FRe exhibits a laxative effect without causing diarrhea, as compared with sodium picosulfate; thus, FRe may be effective as a complementary medicine in patients suffering from lifestyle-induced constipation.

  7. Generation and Analysis of Transposon Ac/Ds-Induced Chromosomal Rearrangements in Rice Plants.

    Science.gov (United States)

    Xuan, Yuan Hu; Peterson, Thomas; Han, Chang-Deok

    2016-01-01

    Closely-located transposable elements (TEs) have been known to induce chromosomal breakage and rearrangements via alternative transposition. To study genome rearrangements in rice, an Ac/Ds system has been employed. This system comprises an immobile Ac element expressed under the control of CaMV 35S promoter, and a modified Ds element. A starter line carried Ac and a single copy of Ds at the OsRLG5 (Oryza sativa receptor-like gene 5). To enhance the transpositional activity, seed-derived calli were cultured and regenerated into plants. Among 270 lines regenerated from the starter, one line was selected that contained a pair of inversely-oriented Ds elements at the OsRLG5 (Oryza sativa receptor-like gene 5). The selected line was again subjected to tissue culture to obtain a regenerant population. Among 300 regenerated plants, 107 (36 %) contained chromosomal rearrangements including deletions, duplications, and inversions of various sizes. From 34 plants, transposition mechanisms leading to such genomic rearrangements were analyzed. The rearrangements were induced by sister chromatid transposition (SCT), homologous recombination (HR), and single chromatid transposition (SLCT). Among them, 22 events (65 %) were found to be transmitted to the next generation. These results demonstrate a great potential of tissue culture regeneration and the Ac/Ds system in understanding alternative transposition mechanisms and in developing chromosome engineering in plants.

  8. Gallic acid induces apoptosis in EGFR-mutant non-small cell lung cancers by accelerating EGFR turnover.

    Science.gov (United States)

    Nam, Boas; Rho, Jin Kyung; Shin, Dong-Myung; Son, Jaekyoung

    2016-10-01

    Gallic acid is a common botanic phenolic compound, which is present in plants and foods worldwide. Gallic acid is implicated in various biological processes such as cell growth and apoptosis. Indeed, gallic acid has been shown to induce apoptosis in many cancer types. However, the molecular mechanisms of gallic acid-induced apoptosis in cancer, particularly lung cancer, are still unclear. Here, we report that gallic acid induces apoptosis in EGFR-mutant non-small cell lung cancer (NSCLC) cells, but not in EGFR-WT NSCLC cells. Treatment with gallic acid resulted in a significant reduction in proliferation and induction of apoptosis, only in EGFR-mutant NSCLC cells. Interestingly, treatment with gallic acid led to a robust decrease in EGFR levels, which is critical for NSCLC survival. Treatment with gallic acid had no significant effect on transcription, but induced EGFR turnover. Indeed, treatment with a proteasome inhibitor dramatically reversed gallic acid-induced EGFR downregulation. Moreover, treatment with gallic acid induced EGFR turnover leading to apoptosis in EGFR-TKI (tyrosine kinase inhibitor)-resistant cell lines, which are dependent on EGFR signaling for survival. Thus, these studies suggest that gallic acid can induce apoptosis in EGFR-dependent lung cancers that are dependent on EGFR for growth and survival via acceleration of EGFR turnover.

  9. Molecular characterization of Tn5-induced symbiotic (Fix-) mutants of Rhizobium meliloti.

    OpenAIRE

    Zimmerman, J L; Szeto, W W; Ausubel, F M

    1983-01-01

    To investigate the expression of specific symbiotic genes during the development of nitrogen-fixing root nodules, we conducted a systematic analysis of nodule-specific proteins and RNAs produced after the inoculation of alfalfa roots with a series of Rhizobium meliloti mutants generated by site-directed transposon Tn5 mutagenesis. The mutagenized region of the Rhizobium genome covered approximately 10 kilobases and included the region encoding the nitrogenase polypeptides. All mutant strains ...

  10. Analysis of Spontaneous,gamma ray-and ethylnitrosourea-induced hprt mutants in HL-60 cells With multiplex PCR

    Institute of Scientific and Technical Information of China (English)

    Sheng-Xue Liu; Jia Cao; Hui An; Hua-Min Shun; Lu-Jun Yang; Yong Liu

    2003-01-01

    AIM: To explore the molecular spectra and mechanism of human hypoxanthine guanine phosphoribosyl transferase (hprt) gene mutation induced by ethyluitrosourea (ENU) and 60Co γ-rays.METHODS: Independent human promyelocytic leukemia cells (HL-60) mutants at the hprt locus were isolated from untreated, ethyluitrosourea (ENU) and 60Co γ-ray-exposed cells, respectively, and verified by two-way screening. The genetic changes underlying the mutation were determined by multiplex polymerase chain reaction (PCR) amplification and electrophoresis technique.RESULTS: With dosage increased, survival rate of plated cell reduced (in the group with dosage of ENU with 100-200μg/ml, P<0.01; in the group with dosage of 60Co γ-ray with 2-4 Gy, P<0.05) and mutational frequency increased (in the group of ENU 12.5-200.0 μg/ml, P<0.05; in the group of 60Co γ-ray with 1-4 Gy, P<0.05) significantly. In the 13spontaneous mutants analyzed, 92.3 % of mutant clones did not show any change in number or size of exon, a single exon was lost in 7.7 %, and no evidence indicated total gene deletion occurred in nine hprt exons. However, deletions were found in 79.7 % of ENU-induced mutations (62.5-89.4 %,P<0.01) and in 61.7 % of gamma-ray-induced mutations (28.6-76.5 %, P<0.01). There were deletion mutations in all 9 exons of hprt gene and the most of induced mutations were chain deletion with multiplex exons (97.9 % in gammaray-induced mutants, 88.1% in ENU-induced mutants).CONCLUSION: The spectra of spontaneous mutations differs completely from that induced by EUN or 60Co γ-ray.Although both ENU and γ-ray can cause destruction of genetic structure, mechanism of mutagenesis between them may be different.

  11. Host-Induced Gene Silencing of Rice Blast Fungus Magnaporthe oryzae Pathogenicity Genes Mediated by the Brome Mosaic Virus.

    Science.gov (United States)

    Zhu, Lin; Zhu, Jian; Liu, Zhixue; Wang, Zhengyi; Zhou, Cheng; Wang, Hong

    2017-09-26

    Magnaportheoryzae is a devastating plant pathogen, which has a detrimental impact on rice production worldwide. Despite its agronomical importance, some newly-emerging pathotypes often overcome race-specific disease resistance rapidly. It is thus desirable to develop a novel strategy for the long-lasting resistance of rice plants to ever-changing fungal pathogens. Brome mosaic virus (BMV)-induced RNA interference (RNAi) has emerged as a useful tool to study host-resistance genes for rice blast protection. Planta-generated silencing of targeted genes inside biotrophic pathogens can be achieved by expression of M.oryzae-derived gene fragments in the BMV-mediated gene silencing system, a technique termed host-induced gene silencing (HIGS). In this study, the effectiveness of BMV-mediated HIGS in M.oryzae was examined by targeting three predicted pathogenicity genes, MoABC1,MoMAC1 and MoPMK1. Systemic generation of fungal gene-specific small interfering RNA (siRNA) molecules induced by inoculation of BMV viral vectors inhibited disease development and reduced the transcription of targeted fungal genes after subsequent M.oryzae inoculation. Combined introduction of fungal gene sequences in sense and antisense orientation mediated by the BMV silencing vectors significantly enhanced the efficiency of this host-generated trans-specific RNAi, implying that these fungal genes played crucial roles in pathogenicity. Collectively, our results indicated that BMV-HIGS system was a great strategy for protecting host plants against the invasion of pathogenic fungi.

  12. Development of functional analysis system for nerve growth factor of gene using {gamma}-ray induced fish mutants

    Energy Technology Data Exchange (ETDEWEB)

    Araki, Kazuo; Nagoya, Hiroyuki; Okamoto, Hiroyuki [National Research Inst. of Agriculture, Mie (Japan)

    2000-02-01

    Development of an effective method to produce {gamma}-ray induced mutants was attempted using medaka, a killifish of which development has been analyzed in detail. An experimental system to make functional analysis for the gene of nerve growth factor was constructed using {gamma} ray -induced mutants and the mechanism to induce its morphogenesis and the nerve growth common to invertebrates was investigated. Some mutants with abnormalities in the chorda dorsalis formation were constructed by exposure to {gamma} ray. Thus, the expression of HNF3{beta} gene which is likely to be involved in the formation of chorda dorsalis was investigated and it was demonstrated that this gene is not expressed in the end region of chorda dorsalis and its succeeding mesoderm. Moreover, the genes expressed at the end region of wild type killifish was isolated by differential hybridization for DNA library that was constructed based on cDNA for the mRNA produced in the range. Twenty genes homologous to the previously reported genes and 51 not homologous ones were isolated. These genes would be utilized for the investigation of their expression. (M.N.)

  13. Scriptaid overcomes hypoxia-induced cisplatin resistance in both wild-type and mutant p53 lung cancer cells.

    Science.gov (United States)

    Pradhan, Shrikant; Mahajan, Divyank; Kaur, Prabhjot; Pandey, Namita; Sharma, Chandresh; Srivastava, Tapasya

    2016-11-01

    Non-small cell lung cancer (NSCLC), comprising 85% of lung cancer cases, has been associated with resistance to chemo/radiotherapy. The hypoxic tumor micro-environment, where insufficient vasculature results in poor drug penetrance and sub-optimal chemotherapy in the tumor interiors contributes heavily to this resistance. Additionally, epigenetic changes in tumorigenic cells also change their response to different forms of therapy. In our study, we have investigated the effectiveness of a combination of cisplatin with scriptaid [a pan-Histone Deacetylase inhibitor (HDACi)] in a model that mimics the tumor microenvironment of hypoxia and sub-lethal chemotherapy. Scriptaid synergistically increases the efficacy of cisplatin in normoxia as well as hypoxia, accompanied with reduced metastasis and enhanced DNA damage. Addition of scriptaid also overcomes the cisplatin resistance exhibited in lung cancer cells with stabilized hypoxia inducible factor 1 (HIF1)-α (mutant) and mutant p53. Molecular studies showed that the combination treatment increased apoptotic cell death in both normoxia and hypoxia with a dual role of p38MAPK. Together, our results suggest that the combination of low dose cisplatin and scriptaid is cytotoxic to NSCLC lines, can overcome hypoxia induced resistance and mutant p53- induced instability often associated with this cancer, and has the potential to be an effective therapeutic modality.

  14. Purification and characterization of a novel hypersensitive response-inducing elicitor from Magnaporthe oryzae that triggers defense response in rice.

    Directory of Open Access Journals (Sweden)

    Mingjia Chen

    Full Text Available BACKGROUND: Magnaporthe oryzae, the rice blast fungus, might secrete certain proteins related to plant-fungal pathogen interactions. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we report the purification, characterization, and gene cloning of a novel hypersensitive response-inducing protein elicitor (MoHrip1 secreted by M. oryzae. The protein fraction was purified and identified by de novo sequencing, and the sequence matched the genomic sequence of a putative protein from M. oryzae strain 70-15 (GenBank accession No. XP_366602.1. The elicitor-encoding gene mohrip1 was isolated; it consisted of a 429 bp cDNA, which encodes a polypeptide of 142 amino acids with a molecular weight of 14.322 kDa and a pI of 4.53. The deduced protein, MoHrip1, was expressed in E. coli. And the expression protein collected from bacterium also forms necrotic lesions in tobacco. MoHrip1 could induce the early events of the defense response, including hydrogen peroxide production, callose deposition, and alkalization of the extracellular medium, in tobacco. Moreover, MoHrip1-treated rice seedlings possessed significantly enhanced systemic resistance to M. oryzae compared to the control seedlings. The real-time PCR results indicated that the expression of some pathogenesis-related genes and genes involved in signal transduction could also be induced by MoHrip1. CONCLUSION/SIGNIFICANCE: The results demonstrate that MoHrip1 triggers defense responses in rice and could be used for controlling rice blast disease.

  15. Traits Related to Chilling-Induced Photoinhibition in Leaves of indica and japonica Rice (Oryza sativa)

    Institute of Scientific and Technical Information of China (English)

    ZHU Su-qin; JI Ben-hua; JIAO De-mao

    2004-01-01

    Physiological indices related to PS Ⅱ photochemical efficiency (Fv/Fm) and membrane lipid peroxidation were measured in leaves of indica rice cv Shanyou 63 and japonica rice 9516 at different temperatures and light intensities for four days. No obvious changes in Fv/Fm and MDA were observed in both indica and japonica rice at moderate temperature and medium PFD,implying neither photoinhibition nor photooxidation happened in these cases. In indica rice either at medium temperature with higher PFD or at lower temperature with medium PFD Fv/Fm dropped obviously with no changes in MDA contents, and photoinhibition appeared while photooxidation did not occur. However, D1 protein, Fv/Fm, (A+Z)/(A+Z+V), and SOD activities dropped, and O2 -. production and MDA content increased accordingly, as well as both photoinhibition and photooxidation appeared in two rice varieties at lower temperature and higher PFD. Experiment with inhibitors at lower temperature and higher PFD showed that as compared with japonica rice the decrements appeared in D1 protein contents, SOD activities, and (A+Z)/(A+Z+V) ratios, the xanthophyll cycle and non-photochemical quench (qN) were inhibited in a more degree, as well as increments of MDA content were greater, thus exhibiting more distinct photoinhibition and photooxidation in indica rice. It is suggested that Fv/Fm and membrane lipid peroxidation product-MDA were the key indices to predict and diagnose photooxidation.

  16. A phosphomimetic mutant TDP-43 (S409/410E) induces Drosha instability and cytotoxicity in Neuro 2A cells.

    Science.gov (United States)

    Kim, Ki Yoon; Lee, Hee-Woo; Shim, Yu-Mi; Mook-Jung, Inhee; Jeon, Gye Sun; Sung, Jung-Joon

    2015-08-14

    Two DNA/RNA binding proteins, TDP-43 and FUS/TLSU, are involved in RNA processing, and their aberrant mutations induce inherited amyotrophic lateral sclerosis and frontotemporal lobar degeneration with ubiquitinated inclusions. Wild type TDP-43 and FUS (wtTDP-43 and wtFUS) are mainly localized in the nucleus and biochemically interact with the microRNA processing enzyme Drosha. In this study, we investigated Drosha stability in Neuro 2A cells by gain and loss of function studies of wtTDP-43 and wtFUS and cycloheximide mediated protein degradation assay. We also generated three different phosphomimetic mutants of TDP-43 (S379E, S403/404E and S409/410E) by using a site-directed mutagenesis method and examined Drosha stability to elucidate a correlation between the phosphorylated TDP-43 mutants and Drosha stability. Overexpression of wtTDP-43 and/or wtFUS increased Drosha stability in Neuro 2A cells and double knockdown of wtTDP-43 and wtFUS reduced its stability. However, knockdown of wtTDP-43 or wtFUS did not affect Drosha stability in Neuro 2A cells. Interestingly, a phosphomimetic mutant TDP-43 (S409/410E) significantly reduced Drosha stability via prevention of protein-protein interactions between wtFUS and Drosha, and induced cytotoxicity in Neuro 2A cells. Our findings suggest that TDP-43 and FUS controls Drosha stability in Neuro 2A cells and that a phosphomimetic mutant TDP-43 (S409/410E) which is associated with Drosha instability can induce neuronal toxicity.

  17. Valproate-induced teratogenesis in Japanese rice fish (Oryzias latipes) embryogenesis.

    Science.gov (United States)

    Wu, Mengmeng; Khan, Ikhlas A; Dasmahapatra, Asok K

    2012-04-01

    Fertilized eggs of Japanese rice fish (medaka) at three developmental stages (Iwamatsu stages 4-30) were exposed to waterborne valproic acid (VPA) (0-80 mM) in hatching solution for 48 h. The amount of valproate to cause 50% mortality (IC(50)) is found to be developmental stage-specific. The embryos were more sensitive to valproate at early stages of development (Iwamatsu stages 4-10) than in the embryos in late stages (Iwamatsu stages 17-30). Valproate exposed embryos have microcephaly and disrupted cardiovasculature with delayed vessel circulation, thrombus formation, and slow heart rate. The hatching efficiency is also reduced by valproate exposure due to developmental delay. The mRNA analysis of nine genes belong to oxidative stress (catalase, gsr, gst), neurogenesis (iro3, wnt1, shh, otx2, nlgn3b) and cell cycle regulation (ccna2) have been done. It was observed that the genes belong to oxidative stress remained unaltered after valproate exposure. However, some of the genes belong to neurogenesis (wnt1,shh, otx2 and nlgn3b) and cell cycle (ccna2) showed developmental stage-specific alteration after valproate exposure. This study indicates that valproate is able to induce some of the phenotypic features which are analogous to human fetal valproate syndrome (FVS). Modulation of genes expressed in neural tissues indicates that this fish can be used to analyze the mechanisms of many neurobehavioral disorders like Autism spectrum disorder (ASD) in human.

  18. Germinated brown rice ameliorates obesity in high-fat diet induced obese rats.

    Science.gov (United States)

    Lim, See Meng; Goh, Yong Meng; Mohtarrudin, Norhafizah; Loh, Su Peng

    2016-05-23

    Germinated brown rice (GBR) is a novel functional food that is high in fiber and bioactive compounds with health-promoting properties. This study aims to evaluate anti-obesity effects of GBR in obese rats fed high-fat diet (HFD). Male Sprague-Dawley rats were fed HFD for 8 weeks to induce obesity. The rats were then administrated with GBR where the source of dietary carbohydrate of HFD was replaced by either 25 % GBR, 50 % GBR or 100 % GBR for another 8 weeks. Changes in anthropometry, dietary status, biochemical parameters and histopathology of liver and adipose tissue were measured. Rats fed with HFD were showed elevation in body weight gain and in white adipose tissue mass compared with rats consumed commercial diet. The GBR administration in 50 % GBR and 100 % GBR were significantly decreased body weight gains and food intakes as well as improved lipid profiles in obese rats. In addition, the administration of GBR  had reduced adiposity by showing declination in white adipose tissue mass, adipocytes size and leptin level concomitantly with a higher ratio of fat excretion into feces. Micro- and macrovesicular steatosis were evidently attenuated in obese rats fed GBR. These findings demonstrated that GBR exhibited anti-obesity effects through suppression of body weight gain and food intake, improvement of lipid profiles and reduction of leptin level and white adipose tissue mass in obese rats fed HFD.

  19. Red Mold Rice Mitigates Oral Carcinogenesis in 7,12-Dimethyl-1,2-Benz[a]anthracene-Induced Oral Carcinogenesis in Hamster

    Directory of Open Access Journals (Sweden)

    Ruei-Lan Tsai

    2011-01-01

    Full Text Available The prevalence of oral tumor has exponentially increased in recent years; however, the effective therapies or prevention strategies are not sufficient. Red mold rice is a traditional Chinese food, and several reports have demonstrated that red mold rice had an anti-tumor effect. However, the possible anti-tumor mechanisms of the red mold rice are unclear. In this study, we examined the anti-tumor effect of red mold rice on 7,12-dimethyl-1,2-benz[a]anthracene (DMBA-induced oral tumor in hamster. The ethanol extract of red mold rice (RMRE treatment significantly decreases the levels of DMBA-induced reactive oxygen species, nitro oxide and prostaglandin E2 than those of the lovastatin-treated group (P < .001. Moreover, RMRE decreases the formation of oral tumor induced by DMBA. Monacolin K, monascin, ankaflavin or other red mold rice metabolites had been reported to decrease inflammation and oxidative stress and exerted anti-tumor effects. Therefore, we evaluated the anti-inflammation and anti-oxidative stress effects of monacolin K, monascin, ankaflavin and citrinin in lipopolysaccharide-treated RAW264.7 cells. We found that RMRE reduced the LPS-induced nitrite levels in RAW264.7 cells better than monacolin K, monascin, ankaflavin or citrinin (P < .05.

  20. Enhanced lipid productivity and photosynthesis efficiency in a Desmodesmus sp. mutant induced by heavy carbon ions.

    Science.gov (United States)

    Hu, Guangrong; Fan, Yong; Zhang, Lei; Yuan, Cheng; Wang, Jufang; Li, Wenjian; Hu, Qiang; Li, Fuli

    2013-01-01

    The unicellular green microalga Desmodesmus sp. S1 can produce more than 50% total lipid of cell dry weight under high light and nitrogen-limitation conditions. After irradiation by heavy (12)C(6+) ion beam of 10, 30, 60, 90 or 120 Gy, followed by screening of resulting mutants on 24-well microplates, more than 500 mutants were obtained. One of those, named D90G-19, exhibited lipid productivity of 0.298 g L(-1)⋅d(-1), 20.6% higher than wild type, likely owing to an improved maximum quantum efficiency (Fv/Fm) of photosynthesis under stress. This work demonstrated that heavy-ion irradiation combined with high-throughput screening is an effective means for trait improvement. The resulting mutant D90G-19 may be used for enhanced lipid production.

  1. Enhanced lipid productivity and photosynthesis efficiency in a Desmodesmus sp. mutant induced by heavy carbon ions.

    Directory of Open Access Journals (Sweden)

    Guangrong Hu

    Full Text Available The unicellular green microalga Desmodesmus sp. S1 can produce more than 50% total lipid of cell dry weight under high light and nitrogen-limitation conditions. After irradiation by heavy (12C(6+ ion beam of 10, 30, 60, 90 or 120 Gy, followed by screening of resulting mutants on 24-well microplates, more than 500 mutants were obtained. One of those, named D90G-19, exhibited lipid productivity of 0.298 g L(-1⋅d(-1, 20.6% higher than wild type, likely owing to an improved maximum quantum efficiency (Fv/Fm of photosynthesis under stress. This work demonstrated that heavy-ion irradiation combined with high-throughput screening is an effective means for trait improvement. The resulting mutant D90G-19 may be used for enhanced lipid production.

  2. Che-1 gene silencing induces osteosarcoma cell apoptosis by inhibiting mutant p53 expression

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Ming; Wang, Dan, E-mail: danwangwdd@163.com; Li, Ning

    2016-04-22

    The transcriptional cofactor Che-1 is an RNA polymerase II (Pol II) which is involved in tumorigenesis, such as breast cancer and multiple myeloma. Che-1 can also regulate mutant p53 expression, which plays roles in many types of cancer. In this study, we aimed to investigate the effects and specific mechanism of Che-1 in the regulation of osteosarcoma (OS) cell growth. We found that Che-1 is highly expressed in several kinds of OS cells compared with osteoblast hFOB1.19 cells. MTT and flow cytometry assays showed that Che-1 depletion by siRNA markedly suppressed MG-63 and U2OS cell proliferation and promoted apoptosis. The chromatin immunoprecipitation (ChIP) assay verified the presence of Che-1 on the p53 promoter in MG-63 and U2OS cells carrying mutant p53. Further studies showed that Che-1 depletion inhibited mutant p53 expression. Notably, our study showed that the loss of Che-1 inhibits proliferation and promotes apoptosis in MG-63 cells by decreasing the level of mutant p53. Therefore, these findings open the possibility that silencing of Che-1 will have therapeutic benefit in OS. - Highlights: • Che-1 is highly expressed in several kinds of OS cells. • Che-1 depletion suppressed MG-63 and U2OS cell growth. • Che-1 is existed in the p53 promoter in MG-63 and U2OS cells. • Che-1 depletion inhibited mutant p53 expression. • Che-1 depletion inhibits cell growth by decreasing the level of mutant p53.

  3. Enhanced Lipid Productivity and Photosynthesis Efficiency in a Desmodesmus sp. Mutant Induced by Heavy Carbon Ions

    OpenAIRE

    Guangrong Hu; Yong Fan; Lei Zhang; Cheng Yuan; Jufang Wang; Wenjian Li; Qiang Hu; Fuli Li

    2013-01-01

    The unicellular green microalga Desmodesmus sp. S1 can produce more than 50% total lipid of cell dry weight under high light and nitrogen-limitation conditions. After irradiation by heavy (12)C(6+) ion beam of 10, 30, 60, 90 or 120 Gy, followed by screening of resulting mutants on 24-well microplates, more than 500 mutants were obtained. One of those, named D90G-19, exhibited lipid productivity of 0.298 g L(-1)⋅d(-1), 20.6% higher than wild type, likely owing to an improved maximum quantum ef...

  4. Comparative proteomics of a tor inducible Aspergillus fumigatus mutant reveals involvement of the Tor kinase in iron regulation.

    Science.gov (United States)

    Baldin, Clara; Valiante, Vito; Krüger, Thomas; Schafferer, Lukas; Haas, Hubertus; Kniemeyer, Olaf; Brakhage, Axel A

    2015-07-01

    The Tor (target of rapamycin) kinase is one of the major regulatory nodes in eukaryotes. Here, we analyzed the Tor kinase in Aspergillus fumigatus, which is the most important airborne fungal pathogen of humans. Because deletion of the single tor gene was apparently lethal, we generated a conditional lethal tor mutant by replacing the endogenous tor gene by the inducible xylp-tor gene cassette. By both 2DE and gel-free LC-MS/MS, we found that Tor controls a variety of proteins involved in nutrient sensing, stress response, cell cycle progression, protein biosynthesis and degradation, but also processes in mitochondria, such as respiration and ornithine metabolism, which is required for siderophore formation. qRT-PCR analyses indicated that mRNA levels of ornithine biosynthesis genes were increased under iron limitation. When tor was repressed, iron regulation was lost. In a deletion mutant of the iron regulator HapX also carrying the xylp-tor cassette, the regulation upon iron deprivation was similar to that of the single tor inducible mutant strain. In line, hapX expression was significantly reduced when tor was repressed. Thus, Tor acts either upstream of HapX or independently of HapX as a repressor of the ornithine biosynthesis genes and thereby regulates the production of siderophores.

  5. Suppressive effect of astaxanthin isolated from the Xanthophyllomyces dendrorhous mutant on ethanol-induced gastric mucosal injury in rats.

    Science.gov (United States)

    Kim, Jeong-Hwan; Choi, Seok-Keun; Choi, Sang-Yun; Kim, Han-Kyeom; Chang, Hyo-Ihl

    2005-07-01

    Ethanol has been found to induce ulcerative gastric lesion in humans. The present study investigated the in vivo protective effect of astaxanthin isolated from the Xanthophyllomyces dendrorhous mutant against ethanol-induced gastric mucosal injury in rats. The rats were treated with 80% ethanol for 3 d after pretreatment with two doses of astaxanthin (5 and 25 mg/kg of body weight respectively) for 3 d, while the control rats received only 80% ethanol for 3 d. The oral administration of astaxanthin (5 and 25 mg/kg of body weight) showed significant protection against ethanol-induced gastric lesion and inhibited elevation of the lipid peroxide level in gastric mucosa. In addition, pretreatment with astaxanthin resulted in a significant increase in the activities of radical scavenging enzymes such as superoxide dismutase, catalase, and glutathione peroxidase. A histologic examination clearly indicated that the acute gastric mucosal lesion induced by ethanol nearly disappeared after pretreatment with astaxanthin.

  6. Changes in Unsaturated Levels of Fatty Acids in Thylakoid PSⅡ Membrane Lipids During Chilling-induced Resistance in Rice

    Institute of Scientific and Technical Information of China (English)

    Su-Qin Zhu; Chun-Mei Yu; Xin-Yan Liu; Ben-Hua Ji; De-Mao Jiao

    2007-01-01

    Temperature is one of the abiotic factors limiting growth and productivity of plants. In the present work, the effect of low non-freezing temperature, as an inducer of "chilling resistance", was studied in three cultivars of rice (Oryza sativa L.), japonica cv. 9516 (j-9516), the two parental lines of superhigh-yield hybrid rice between subspecies,Peiai/E32 (ji-PE), and the traditional indica hybrid rice Shanyou 63 (i-SY63). Leaves of chill-treated rice showed chilling-induced resistance, as an increase of their low-temperature tolerance was measured using chlorophyll fluorescence measurements, revealing a change in photosystem Ⅱ (PSⅡ) efficiency. After 5 d of exposure to 11℃ under low light (100 μ mol·m-2·s-1), levels of unsaturated fatty acids in PSⅡ thylakoid membrane lipids decreased during the initial 1-2 d, then increased slowly and reached 99.2%, 95.3% and 90.1% of the initial value (0 d) in j-9516,ji-PE and i-SY63, respectively, on the fifth day. However, under medium light (600 μmol·m-2·s-1), all cultivars experienced similar substantial photoinhibition, which approached steady state levels after a decline in levels of unsaturated fatty acids in PSII thylakoid membrane lipids to about 57.1%, 53.8% and 44.5% of the initial values (0 d) in j-9516,ji-PE and i-SY63 on the fifth day. Under either chilling-induced resistance (the former) or low temperature photoinhibition (the latter) conditions, the changes of other physiological parameters such as D1 protein contents,electron transport activities of PSII (ETA), Fv/Fm, xanthophyl cycle activities expressed by DES (deepoxide state)were consistent with that of levels of unsaturated fatty acids in PSⅡ thylakoid membrane lipids. So there were negative correlations between saturated levels of fatty acids (16:1(3t), 16:0, 18:0), especially the 16:1(3t) fatty acid on thylakoid membrane and other physiological parameters, such as D1 protein contents, ETA and (A+Z)/(A+V+Z). A specific role of

  7. Heavy-ion radiation induces both activation of multiple endogenous transposable elements and alterations in DNA methylation in rice

    Science.gov (United States)

    Zhang, Meng; Sun, Yeqing; Li, Xishan; Xiaolin, Cui; Li, Xiang

    2012-07-01

    Space radiation represents a complex environmental condition in which several interacting factors such as electron, neutron, proton, heavy-ion are involved, which may provoke stress responses and jeopardize genome integrity. Given the inherent property of epigenetic modifications to respond to intrinsic aswell as external perturbations, it is conceivable that epigenetic markers like DNA methylation and transposition may undergo alterations in response to space radiation. Cytosine DNA methylation plays important roles in maintaining genome stability and controlling gene expression. A predominant means for Transposable elements (TEs) to cause genetic instability is via their transpositional activation. To find the detailed molecular characterization of the nature of genomic changes induced by space radiation, the seeds of rice were exposed to 0.02, 0.2, 1, 2 and 20 Gy dose of ^{12}C heavy-ion radiation, respectively. We found that extensive alteration in both DNA methylation and gene expression occurred in rice plants after different dose of heavy-ion radiation. Here we shown that heavy-ion radiation has induced transposition of mPing and Tos17 in rice, which belong to distinct classes including the miniature inverted terminal repeat TEs (MITEs) and long-terminal repeat (LTR) retrotransposons, respectively. mPing and Tos17 mobility were found to correlate with cytosine methylation alteration detected by MSAP and genetic variation detected by AFLP. The result showed that at least in some cases transposition of TEs was associated with cytosine demethylation within the elements. Our results implicate that the heavy-ion radiation represents a potent mutagenic agent that can cause genomic instabilities by eliciting transposition of endogenous TEs in rice. Keywords: Heavy-ion radiation, DNA methylation, Transposable elements, mPing, Tos17

  8. Gamma ray-induced mutants as a tool for the production and characterisation of monoclonal antibodies against HLA-alloantigens

    Energy Technology Data Exchange (ETDEWEB)

    Spring, B.; Pawelec, G.; Ziegler, A.

    1986-01-01

    To simplify the screening procedure for murine monoclonal antibodies specific for polymorphic HLA determinants, spleen cells from a mouse immunized with the human cell line BJAB-B95.8.6 were fused with NS1 mouse myeloma cells, and hybridoma supernatants were screened for their reactivity on BJAB-B95.8.6 and two gamma ray-induced HLA-loss mutants of this line. The use of these HLA-loss mutants allowed the rapid identification of two new allospecific MOABs designated TU160 and TU161. Serological as well as biochemical studies revealed TU160 to be specific for HLA=A2, and TU161 for HLA-B13 molecules, respectively. Both MOABs were determined to be antibodies of the IgG class and were able to precipitate their antigens from lysates of radioactively labeled cells.

  9. Evaluation of Genetic Variation Induced by Gamma Ray Irradiation in Morphological Traits in M2 Mutant Lines of Canola

    Directory of Open Access Journals (Sweden)

    S. M. AbtahiForushani

    2014-09-01

    Full Text Available Mutation induction is considered as an effective way to enrich plant genetic variation, particularly for traits with a very low level of genetic variation. The objectives of this study were to evaluate the effect of 1200 GY gamma ray irradiation on morphophenological traits, yield and yield components and to identify useful mutants in two spring rapeseed "RGS003" and" Sarigol" cultivars. The results revealed that there are highly significant difference sa mong mutant lines. Based on the results of phenotypic and genetic correlations, 1000 seed weight had the highest relationship with see dyieldper plant. Stepwise regression analysis also showed that 1000 seed weight was one of the most important traits contributing in seed yield.In viewpoint of the exploitation of the induced variation at morphological traits, M2 lines 9 from RGS003 and 16, 25 and 26 from Sarigol could be suggested as the most desirable lines for canola breeding.

  10. Functional Inactivation of Putative Photosynthetic Electron Acceptor Ferredoxin C2 (FdC2) Induces Delayed Heading Date and Decreased Photosynthetic Rate in Rice.

    Science.gov (United States)

    Zhao, Juan; Qiu, Zhennan; Ruan, Banpu; Kang, Shujing; He, Lei; Zhang, Sen; Dong, Guojun; Hu, Jiang; Zeng, Dali; Zhang, Guangheng; Gao, Zhenyu; Ren, Deyong; Hu, Xingming; Chen, Guang; Guo, Longbiao; Qian, Qian; Zhu, Li

    2015-01-01

    Ferredoxin (Fd) protein as unique electron acceptor, involved in a variety of fundamental metabolic and signaling processes, which is indispensable for plant growth. The molecular mechanisms of Fd such as regulation of electron partitioning, impact of photosynthetic rate and involvement in the carbon fixing remain elusive in rice. Here we reported a heading date delay and yellowish leaf 1 (hdy1) mutant derived from Japonica rice cultivar "Nipponbare" subjected to EMS treatment. In the paddy field, the hdy1 mutant appeared at a significantly late heading date and had yellow-green leaves during the whole growth stage. Further investigation indicated that the abnormal phenotype of hdy1 was connected with depressed pigment content and photosynthetic rate. Genetic analysis results showed that the hdy1 mutant phenotype was caused by a single recessive nuclear gene mutation. Map-based cloning revealed that OsHDY1 is located on chromosome 3 and encodes an ortholog of the AtFdC2 gene. Complementation and overexpression, transgenic plants exhibited the mutant phenotype including head date, leaf color and the transcription levels of the FdC2 were completely rescued by transformation with OsHDY1. Real-time PCR revealed that the expression product of OsHDY1 was detected in almost all of the organs except root, whereas highest expression levels were observed in seeding new leaves. The lower expression levels of HDY1 and content of iron were detected in hdy1 than WT's. The FdC2::GFP was detected in the chloroplasts of rice. Real-time PCR results showed that the expression of many photosynthetic electron transfer related genes in hdy1 were higher than WT. Our results suggest that OsFdC2 plays an important role in photosynthetic rate and development of heading date by regulating electron transfer and chlorophyll content in rice.

  11. Functional Inactivation of Putative Photosynthetic Electron Acceptor Ferredoxin C2 (FdC2 Induces Delayed Heading Date and Decreased Photosynthetic Rate in Rice.

    Directory of Open Access Journals (Sweden)

    Juan Zhao

    Full Text Available Ferredoxin (Fd protein as unique electron acceptor, involved in a variety of fundamental metabolic and signaling processes, which is indispensable for plant growth. The molecular mechanisms of Fd such as regulation of electron partitioning, impact of photosynthetic rate and involvement in the carbon fixing remain elusive in rice. Here we reported a heading date delay and yellowish leaf 1 (hdy1 mutant derived from Japonica rice cultivar "Nipponbare" subjected to EMS treatment. In the paddy field, the hdy1 mutant appeared at a significantly late heading date and had yellow-green leaves during the whole growth stage. Further investigation indicated that the abnormal phenotype of hdy1 was connected with depressed pigment content and photosynthetic rate. Genetic analysis results showed that the hdy1 mutant phenotype was caused by a single recessive nuclear gene mutation. Map-based cloning revealed that OsHDY1 is located on chromosome 3 and encodes an ortholog of the AtFdC2 gene. Complementation and overexpression, transgenic plants exhibited the mutant phenotype including head date, leaf color and the transcription levels of the FdC2 were completely rescued by transformation with OsHDY1. Real-time PCR revealed that the expression product of OsHDY1 was detected in almost all of the organs except root, whereas highest expression levels were observed in seeding new leaves. The lower expression levels of HDY1 and content of iron were detected in hdy1 than WT's. The FdC2::GFP was detected in the chloroplasts of rice. Real-time PCR results showed that the expression of many photosynthetic electron transfer related genes in hdy1 were higher than WT. Our results suggest that OsFdC2 plays an important role in photosynthetic rate and development of heading date by regulating electron transfer and chlorophyll content in rice.

  12. Induced drought tolerance through wild and mutant bacterial strain Pseudomonas simiae in mung bean (Vigna radiata L.).

    Science.gov (United States)

    Kumari, Sarita; Vaishnav, Anukool; Jain, Shekhar; Varma, Ajit; Choudhary, Devendra Kumar

    2016-01-01

    The present study focused on the overproducing mutant of a plant growth promoting rhizobacterium (PGPR) Pseudomonas simiae strain AU (MTCC-12057) for significant drought tolerance in mung bean plants. Five mutants namely AU-M1, AU-M2, AU-M3, AU-M4 and AU-M5 were made after treatment of wild type strain with N-methyl-N-nitro-N-nitrosoguanidine. Mutant strain AU-M4 was recorded for enhanced ACC deaminase (ACC-D) activity, indole acetic acid (IAA) production and inorganic phosphate (Pi) solubilization compared to wild strain and other four mutant strains under drought condition. AU-M4 showed higher phosphate solubilization index (8.17) together with higher ACC-D activity (98 nmol/mg/h) and IAA concentration (69.35 µg/ml) compared with the wild type P. simiae strain AU ACC-D activity (79 nmol/mg/h) and IAA concentration (38.98 µg/ml) respectively. In this report, we investigated the effect of both wild and mutant type bacterial strain on mung bean plants under drought stress. Results showed that mutant AU-M4 and wild type strain AU inoculated plants exhibited superior tolerance against drought stress, as shown by their enhanced plant biomass (fresh weight), higher water content, higher proline accumulation and lower osmotic stress injury. Mutant AU-M4 and wild strain AU inoculated plants reduced the ethylene level by 59 and 45% respectively, compared to the control under stress condition. Furthermore, bacterial inoculated plants showed enhanced induced systemic drought tolerance by reducing stomata size and net photosynthesis resulting higher water content in mung bean plants that may help in survival of plants during drought condition. To mitigate the effects of drought stress, use of PGPR will be needed to ensure sufficient production of food from crop plants. Taking current leads available, concerted future research is needed in this area, particularly on field evaluation with application of potential microorganisms.

  13. RNAI INDUCED WING MODIFICATION IN LEON MUTANT DROSOPHILA: A DEVELOPMENTAL ANALYSIS

    Directory of Open Access Journals (Sweden)

    Sandeep Satapathy

    2013-01-01

    Full Text Available The precision of growth of an animal is meticulously regulated by extrinsic and intrinsic factors, with focus on maintenance of organismal homeostasis. The clue to change in physiology or metabolism of an organism, at times can be derived from the changes in phenotypes. In the Drosophila melanogaster model system, GAL 4-overexpressed RNAi driver males (Mini-White Marker, targeted against specific genes, when crossed with Leon mutant (19-2/TM6B females, yield progeny of different wing types. Different RNAi lines expressing the phenotypes in a gradient of sodden, mid to normal; explains the varying severity of the wing phenotypes. The comparison of flies co-expressed RNAi and Leon mutant with wild type or Leon mutant females; show changes in wing phenotype; in terms of wing venation, Anterior Cortical Vein (ACV position, Posterior Cortical Vein (PCV position, bristles on the wing margins and the inter-segmental distance. There is a distinct evidence of both rescue and deterioration phenotype observed at various levels, with the varying levels of RNAi expression in sodden, mid and normal type. A correlational study of these modified wing phenotypes to the physiological and metabolic functionalities; reveals the expression of most of these genes targeted by RNAi, mainly in the brain, heart, thoracic-abdominal ganglion, salivary gland, ovary and testis. Therefore, it can be hypothesized that the Leon mutant can be correlated with the RNAi.

  14. Induced protein polymorphisms and nutritional quality of gamma irradiation mutants of sorghum

    CSIR Research Space (South Africa)

    Mehlo, L

    2013-09-01

    Full Text Available by an unusually high level accumulation of free lysine and other essential amino acids in the endosperm. This mutant further displayed a significant suppression in the synthesis and accumulation of the 27 kDa -, 24 kDa a-A1 and the 22 kDa a-A2 kafirins...

  15. Identification of radiation induced mutants of cassava (Manihot esculenta Crantz using morphological and physicochemical descriptors1

    Directory of Open Access Journals (Sweden)

    Tofino Adriana

    2011-08-01

    Full Text Available For mutants of starch and other desirable traits of interest in cassava, we evaluated the physical-chemical morphology of the root and paste in inbred populations of six M2 families, derived from irradiated seeds. 1,097 plants were established, 829 of Gamma treatment and 268 of neutrons. In the morphological description, most chimeras recorded in M1 were identified except hermaphrodite flowers and bark of different colors on the same plant, new abnormalities in M2 were found, such as root without suberification, no anthesis, early flowering, meristems and changes in the supply-demand relationship. However, mortality, reduced vigor, yield and anthesis loss have also been identified as effects of inbreeding. Simple techniques such as optical microscopy and iodine staining were effective for the identification of mutants of the granule and the results were confirmed with electron microscopy analysis and pulp properties, except in mutant granule hole, in which no abnormalities of the granule were correlated with significant variations in the paste. However, Pearson correlations associated the hot paste viscosity with consistency and reorganization. Variability was found in the granule characteristics and properties of the paste between years, suggesting significant influence of genotype-environment interaction. Similarly, there were differences in paste properties associated with the type of radiation. In addition, promising mutants were identified with post-harvest physiological deterioration tolerance associated mainly to gamma.

  16. Passage of Wolbachia pipientis through mutant drosophila melanogaster induces phenotypic and genomic changes.

    Science.gov (United States)

    Newton, Irene L G; Sheehan, Kathy B

    2015-02-01

    Wolbachia pipientis is a nearly ubiquitous, maternally transmitted bacterium that infects the germ line of insect hosts. Estimates are that Wolbachia infects 40 to 60% of insect species on the planet, making it one of the most prevalent infections on Earth. However, we know surprisingly little about the molecular mechanisms used by Wolbachia to infect its hosts. We passaged Wolbachia through normally restrictive Drosophila melanogaster hosts, bottlenecking Wolbachia through stochastic segregation while simultaneously selecting for mutants that could recolonize these previously restrictive hosts. Here, we show that Wolbachia alters its behavior when passaged through heterozygous mutant flies. After only three generations, Wolbachia was able to colonize the previously restrictive hosts at control titers. Additionally, the Wolbachia organisms passaged through heterozygous mutant D. melanogaster alter their pattern of tissue-specific Wsp protein production, suggesting a behavioral response to the host genotype. Using whole-genome resequencing, we identified the mutations accumulated by these lineages of Wolbachia and confirmed the existence and persistence of the mutations through clone library Sanger sequencing. Our results suggest that Wolbachia can quickly adapt to new host contexts, with genomic mutants arising after only two generations.

  17. 5-Fluorouracil preferentially sensitizes mutant KRAS non-small cell lung carcinoma cells to TRAIL-induced apoptosis.

    Science.gov (United States)

    Wang, Haizhen; Yang, Tao; Wu, Xiangwei

    2015-11-01

    Mutations in the KRAS gene are very common in non-small cell lung cancer (NSCLC), but effective therapies targeting KRAS have yet to be developed. Interest in tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), a potent inducer of cell death, has increased following the observation that TRAIL can selectively kill a wide variety of human cancer cells without killing normal cells both in vitro and in xenograft models. However, results from clinical trials of TRAIL-based therapy are disappointingly modest at best and many have demonstrated a lack of therapeutic benefit. Current research has focused on selecting a subpopulation of cancer patients who may benefit from TRAIL-based therapy and identifying best drugs to work with TRAIL. In the current study, we found that NSCLC cells with a KRAS mutation were highly sensitive to treatment with TRAIL and 5-fluorouracil (5FU). Compared with other chemotherapeutic agents, 5FU displayed the highest synergy with TRAIL in inducing apoptosis in mutant KRAS NSCLC cells. We also found that, on a mechanistic level, 5FU preferentially repressed survivin expression and induced expression of TRAIL death receptor 5 to sensitize NSCLC cells to TRAIL. The combination of low-dose 5FU and TRAIL strongly inhibited xenograft tumor growth in mice. Our results suggest that the combination of TRAIL and 5FU may be beneficial for patients with mutant KRAS NSCLC.

  18. Functional analysis of the rice rubisco activase promoter in transgenic Arabidopsis

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Zhipan; Lu, Qingtao; Wen, Xiaogang [Key Laboratory of Photobiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093 (China); Chen, Fan [Key Laboratory of Molecular and Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100080 (China); Lu, Congming, E-mail: lucm@ibcas.ac.cn [Key Laboratory of Photobiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093 (China)

    2012-02-17

    Highlights: Black-Right-Pointing-Pointer Rice rubisco activase promoter was analyzed in transgenic Arabidopsis system. Black-Right-Pointing-Pointer Region conferring tissue specific and light inducible expression of Rca was identified. Black-Right-Pointing-Pointer -58 to +43 bp region mediates tissue-specific expression of rice Rca. Black-Right-Pointing-Pointer Light inducible expression of rice Rca is mediated by -297 to -58 bp region. Black-Right-Pointing-Pointer Rice nuclear proteins bind specifically with the light inducible region. -- Abstract: To gain a better understanding of the regulatory mechanism of the rice rubisco activase (Rca) gene, variants of the Rca gene promoter (one full-length and four deletion mutants) fused to the coding region of the bacterial reporter gene {beta}-glucuronidase (GUS) were introduced into Arabidopsis via Agrobacterium-mediated transformation. Our results show that a 340 bp fragment spanning from -297 to +43 bp relative to the transcription initiation site is enough to promote tissue-specific and light-inducible expression of the rice Rca gene as done by the full-length promoter (-1428 to +43 bp). Further deletion analysis indicated that the region conferring tissue-specificity of Rca expression is localized within a 105 bp fragment from -58 to +43 bp, while light-inducible expression of Rca is mediated by the region from -297 to -58 bp. Gel shift assays and competition experiments demonstrated that rice nuclear proteins bind specifically with the fragment conferring light responsiveness at more than one binding site. This implies that multiple cis-elements may be involved in light-induced expression of the rice Rca gene. These works provide a useful reference for understanding transcriptional regulation mechanism of the rice Rca gene, and lay a strong foundation for further detection of related cis-elements and trans-factors.

  19. Biosynthesis and emission of insect-induced methyl salicylate and methyl benzoate from rice

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, Nan [University of Tennessee, Knoxville (UTK); Guan, Ju [University of Tennessee, Knoxville (UTK); Ferrer, Jean-Luc [Universite Joseph Fourier, France; Engle, Nancy L [ORNL; Chern, Mawsheng [University of California, Davis; Ronald, Pamela [University of California, Davis; Tschaplinski, Timothy J [ORNL; Chen, Feng [University of Tennessee, Knoxville (UTK)

    2010-01-01

    Two benzenoid esters, methyl salicylate (MeSA) and methyl benzoate (MeBA), were detected from insect-damaged rice plants. By correlating metabolite production with gene expression analysis, five candidate genes encoding putative carboxyl methyltransferases were identified. Enzymatic assays with Escherichia coli-expressed recombinant proteins demonstrated that only one of the five candidates, OsBSMT1, has salicylic acid (SA) methyltransferase (SAMT) and benzoic acid (BA) methyltransferase (BAMT) activities for producing MeSA and MeBA, respectively. Whereas OsBSMT1 is phylogenetically relatively distant from dicot SAMTs, the three-dimensional structure of OsBSMT1, which was determined using homology-based structural modeling, is highly similar to those of characterized SAMTs. Analyses of OsBSMT1 expression in wild-type rice plants under various stress conditions indicate that the jasmonic acid (JA) signaling pathway plays a critical role in regulating the production and emission of MeSA in rice. Further analysis using transgenic rice plants overexpressing NH1, a key component of the SA signaling pathway in rice, suggests that the SA signaling pathway also plays an important role in governing OsBSMT1 expression and emission of its products, probably through a crosstalk with the JA signaling pathway. The role of the volatile products of OsBSMT1, MeSA and MeBA, in rice defense against insect herbivory is discussed.

  20. Rice Hypersensitive Induced Reaction Protein 1 (OsHIR1 associates with plasma membrane and triggers hypersensitive cell death

    Directory of Open Access Journals (Sweden)

    Sun Sai-Ming

    2010-12-01

    Full Text Available Abstract Background In plants, HIR (Hypersensitive Induced Reaction proteins, members of the PID (Proliferation, Ion and Death superfamily, have been shown to play a part in the development of spontaneous hypersensitive response lesions in leaves, in reaction to pathogen attacks. The levels of HIR proteins were shown to correlate with localized host cell deaths and defense responses in maize and barley. However, not much was known about the HIR proteins in rice. Since rice is an important cereal crop consumed by more than 50% of the populations in Asia and Africa, it is crucial to understand the mechanisms of disease responses in this plant. We previously identified the rice HIR1 (OsHIR1 as an interacting partner of the OsLRR1 (rice Leucine-Rich Repeat protein 1. Here we show that OsHIR1 triggers hypersensitive cell death and its localization to the plasma membrane is enhanced by OsLRR1. Result Through electron microscopy studies using wild type rice plants, OsHIR1 was found to mainly localize to the plasma membrane, with a minor portion localized to the tonoplast. Moreover, the plasma membrane localization of OsHIR1 was enhanced in transgenic rice plants overexpressing its interacting protein partner, OsLRR1. Co-localization of OsHIR1 and OsLRR1 to the plasma membrane was confirmed by double-labeling electron microscopy. Pathogen inoculation studies using transgenic Arabidopsis thaliana expressing either OsHIR1 or OsLRR1 showed that both transgenic lines exhibited increased resistance toward the bacterial pathogen Pseudomonas syringae pv. tomato DC3000. However, OsHIR1 transgenic plants produced more extensive spontaneous hypersensitive response lesions and contained lower titers of the invading pathogen, when compared to OsLRR1 transgenic plants. Conclusion The OsHIR1 protein is mainly localized to the plasma membrane, and its subcellular localization in that compartment is enhanced by OsLRR1. The expression of OsHIR1 may sensitize the plant

  1. 水稻体细胞无性系雄性不育突变%Male Sterile Mutants from Rice Somaclones

    Institute of Scientific and Technical Information of China (English)

    范树国; 梁承邺; 刘鸿先

    2000-01-01

    对5个类型的水稻不育系进行了幼穗培养,在红源A、包源A和W6154s中,共获得了10例雄性不育变异株,水稻花粉败育可分为无花粉、典败、圆败和染败四种类型。发现了不育花粉败育类型之间可以相互转换现象。对雄性不育变异株用一批现有CMS不育系的保持系和恢复系进行测交和回交,有的变异株其恢保关系发生了变化,由包台型变成了野败型;有的变异株找不到恢复系;而有的变异株则找不到保持系,表现为核不育的性质。%Young panicles from fiva rice varieties,Zhenshan 97A(WA type cytoplasmic malesterile(CMS)line,indica),Hongyuan A(HL type CMS line,indica),Baoyuan A(BT type CMS line,indica),Nongken 58s(photoperiod-sensitive genic male-sterile rice(PGMR),japonica)and W6154s(thermo-sensitive genic male-sterile rice(TGMR),indica),were cultured on the other was coded S14-1, TAT) and Baoyuan A (one was coded 28-5, SpAT; the other wascoded 28-19, TAT), kept sterile continuously for five generations when they were backcrossedwith the ten parents stated above.The other five male-sterile variants, which derived from Hongyuan A (coded 24-5),Baoyuan A(coded 28-35) and W6154s(coded WP-l, WP-2 and WS) turned into fertility with alot of fertile pollens and very high self seed set rate (more than 80%) when they were crossed with the ten parents stated above.different media. After obtained regenerating plants,examined pollen fertility,madetestcrosses of male-sterile variants,did genetic analyses of male-sterile variants,discussedrestoring and maintaining relationship of male sterile variants,we found 10 cases of male-sterilevariants in three rice variedes,Hongyuan A,Baoyuan A and W6154s.Abortive pollen of ricecould be classified into four types:pollen free type(PFT),typical abortion type (TAT),pherical abortion type(SpAT)and stained abortion type(StAT).The phenomenon that type of polln abortion

  2. Analysis on the DNA Fingerprinting of Aspergillus Oryzae Mutant Induced by High Hydrostatic Pressure

    Institute of Scientific and Technical Information of China (English)

    WANG Hua; ZHANG Jian; YANG Fan; WANG Kai; SHEN Si-Le; LIU Bing-Bing; ZOU Bo; ZOU Guang-Tian

    2011-01-01

    The mutant strains of aspergillus oryzae (HP300a) are screened under 300 Mpa for 20min. Compared with the control strains, the screened mutant strains have unique properties such as genetic stability, rapid growth, lots of spores, and high protease activity. Random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) are used to analyze the DNA fingerprinting of HP300a and the control strains. There are 67.9% and 51.3% polymorphic bands obtained by these two markers, respectively, indicating significant genetic variations between HP300a and the control strains. In addition, comparison of HP300a and the control strains, the genetic distances of random sequence and simple sequence repeat of DNA are 0.51 and 0.34, respectively.

  3. 水稻雄性不育突变体012S-3的遗传分析和基因定位%Genetic Analysis and Gene Mapping of a Male Sterile Mutant 012S-3 in Rice

    Institute of Scientific and Technical Information of China (English)

    欧阳杰; 王楚桃; 朱子超; 何永歆; 蒋刚; 黄乾龙; 管玉圣; 熊英; 李贤勇

    2015-01-01

    Male sterility is an important resource for utilization of Heterosis in rice. Research on male sterility has important theoretical significance and practical value. In this study, we investigated the phenotypic characteristics and pollen fertility of the rice natural male-sterile mutant 012S-3. The genetic populations were constructed for preliminary positioning of the target gene, and genome sequencing technology were used for fine positioning of the, target gene. The results showed that 012S-3 was a typical non-pollen type mutant of male sterility and its sterility was controlled by a pairs of recessive nuclear genes. Preliminary mapping analysis indicated the objective gene and SSR marker RM6081 linkage, and their genetic distance was about 34.4 cM. By further fine mapping analysis, three candidate genes were found: LOC_Os07g35880, LOC_Os07g35920 and LOC_Os07g35940. In which, LOC_Os07g35880 and LOC_Os07g35940 encoding two putative β-amylases, belong to the new pollen lethal genes in rice. The successful positioning of the sterile gene laid a foundation for its further isolation, cloning and its application in rice molecular design breeding.%雄性不育是水稻杂种优势利用的重要资源,对雄性不育现象的研究具有重要理论意义和实践价值。本研究以自然突变的水稻雄性不育突变体012S-3为试验材料,对其表型特征和花粉育性等进行调查,并构建遗传群体,利用分子标记对目的基因进行初步定位,然后应用基因组重测序技术对其进行精细定位。结果表明,012S-3是一个典型的无花粉普通型雄性不育材料,其不育性状受1对隐性核基因控制。初步定位分析目的基因与SSR标记RM6081存在连锁关系,其遗传距离约为34.4 cM;进一步的精细定位分析,找到3个候选基因:LOC_Os07g35880、LOC_Os07g35920和LOC_Os07g35940,其中LOC_Os07g35880和LOC_Os07g35940编码β-淀粉酶,属于水稻中新发现的花粉致死基因

  4. Comparative mapping of QTLs for AI tolerance in rice and identification of positional Al-induced genes

    Institute of Scientific and Technical Information of China (English)

    毛传澡; 杨玲; 郑炳松; 吴运荣; 刘非燕; 易可可; 吴平

    2004-01-01

    Aluminum (A1) toxicity is the major factor limiting crop productivity in acid soils. In this study, a recombinant inbreed line (RIL) population derived from a cross between an A1 sensitive lowland indica rice variety IR1552 and an A1 tolerant upland japonica rice variety Azucena, was used for mapping quantitative trait loci (QTLs) for A1 tolerance. Three QTLs for relative root length (RRL) were detected on chromosome 1,9, 12, respectively, and 1 QTL for root length under A1 stress is identical on chromosome 1 after one week and two weeks stress. Comparison of QTLs on chromosome 1 from different studies indicated an identical interval between C86 and RZ801 with gene(s) for A1 tolerance. This interval provides an important start point for isolating genes responsible for A1 tolerance and understanding the genetic nature of Al tolerance in rice. Four A1 induced ESTs located in this interval were screened by reverse Northern analysis and confirmed by Northern analysis. They would be candidate genes for the QTL.

  5. Comparative mapping of QTLs for Al tolerance in rice and identification of positional Al-induced genes

    Institute of Scientific and Technical Information of China (English)

    毛传澡; 杨玲; 郑炳松; 吴运荣; 刘非燕; 易可可; 吴平

    2004-01-01

    Aluminum (Al) toxicity is the major factor limiting crop productivity in acid soils. In this study, a recombinant inbreed line (RIL) population derived from a cross between an A1 sensitive lowland indica rice variety IR1552 and an Al tolerant upland japonica rice variety Azucena, was used for mapping quantitative trait loci (QTLs) for A1 tolerance. Three QTLs for relative root length (RRL) were detected on chromosome 1, 9, 12, respectively, and I QTL for root length under Al stress is identical on chromosome I after one week and two weeks stress. Comparison of QTLs on chromosome 1 from different studies indicated an identical interval between C86 and RZ801 with gene(s) for Al tolerance. This interval provides an important start point for isolating genes responsible for A1 tolerance and understanding the genetic nature of Al tolerance in rice. Four Al induced ESTs located in this interval were screened by reverse Northern analysis and confirmed by Northern analysis. They would be candidate genes for the QTL.

  6. Investigating the roles of MicroRNAs in biotic stress response induced by Rhizoctonia solani in rice

    Energy Technology Data Exchange (ETDEWEB)

    Syuhada, O. Nurfarahana; Kalaivani, N. [School of Environmental and Natural Resource Sciences, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor (Malaysia)

    2014-09-03

    Sheath blight disease, caused by Rhizoctonia solani 1802/KB was screened on two rice varieties, Oryza sativaindica cultivar MR219 and Oryza sativa indica cultivar UKMRC9. The disease symptom was severe in MR219 compared to UKMRC9. Total RNA from R. solani 1802/KB, infected rice leaves of MR219 and infected rice leaves of UKMRC9 were extracted using TRIzol reagent, purified and sent for small RNA sequencing. Three miRNA libraries were generated and analyzed. The libraries generated 65 805, 78 512 and 81 325 known miRNAs respectively. The structure of miRNA of these samples was predicted. The up-regulated and down-regulated of miRNAs target gene prediction and its target functions were discovered and were mainly related to the growth and development of metabolism, protein transport, transcriptional regulation, stress response, and hormone signaling and electron transfer. Sheath blight-induced differential expression of known miRNAs tends to targetMYB transcription factor, F-box proteins, NBS-LRR, leucine-rich repeat receptor protein kinases and zinc finger proteins. Detecting new miRNAs and measuring the expression profiles of known miRNAs is an important tasks required for a better understanding of various biological conditions. Therefore, further analysis using Gene Ontology Slim will be conducted to deduce some biological information from the datasets obtained.

  7. Inducible expression pattern of rice Bowman-Birk inhibitor gene Os WIP1-2 and its protease inhibitory activity

    Institute of Scientific and Technical Information of China (English)

    CHEN Jun; LIU Jing; GUO Lei; QU Lijia; CHEN Zhangliang; GU Hongya

    2004-01-01

    The WIP1-2 gene was cloned from rice. It belongs to the Bowman-Birk inhibitor gene family. Northern blot showed that expression of this gene was induced by wounding and jasmonic acid (JA). It indicates that the OsWIP1 gene plays an important role in the rice defense system. The OsWIP1-2 was cloned into pET28a and expressed in E. Coli. Its expressed product was purified in the form of fusion protein and tested for the inhibitory activities against trypsin and chymotrypsin. It was found that the fusion protein could inhibit chymotrypsin, but not trypsin. It was also found that the His tag at its C-terminal affected its inhibitory activity significantly. The fusion protein with a natural C-terminal had the inhibitory activity, while no inhibitory activity was detected in the fusion protein with a (His)6-tag at its C-terminal. This implies that extra amino acid residues at the C-terminal of OsWIP1-2 may interfere with its correct folding. The inhibitory assay indicated that the members of rice Bowman-Birk inhibitor gene family probably differentiated both in their structure and function.

  8. Soaking Induced Changes in Chemical Composition, Glycemic Index and Starch Characteristics of Basmati Rice

    Directory of Open Access Journals (Sweden)

    S.J. Kale

    2015-09-01

    Full Text Available An attempt was made to determine the qualitative changes in basmati rice (Pusa Basmati 1121, PB1121 during soaking at 40 °C to 80 °C. Soaking temperature had significant effect (α = 0.01 on chemical composition, glycemic index and starch characteristics of rice. Starch content, apparent amylose content, crude protein content and crude fat content in un-soaked rice were found to be 73.24%, 27.26%, 8.79% and 2.56%, respectively, but differences in these traits were observed after soaking. Amylose to amylopectin ratio (Am/Ap decreased from 0.59 to 0.52 (soaked at 80 °C. Crude fibre and crude ash contents increased after soaking. The mineral composition (K, P, S, Ca, Mg, Mn, Fe, Cu and Zn in soaked rice was found to be 16.46% higher than un-soaked rice at the same degree of polishing. Glycemic index of un-soaked rice was found to be 58.41, but decreased to 54.31 after soaking at 80 °C. Pasting properties, scanning electron microscope images, and X-ray diffractograms suggested partial gelatinization of starch in the temperature range of 60 °C to 80 °C. Based on qualitative changes in rice (apparent amylose content, Am/Ap ratio and crystallinity rate, it was concluded that intermediate soaking temperatures (60 °C to 70 °C would be useful for soaking of PB1121.

  9. Mutant GDF5 enhances ameloblast differentiation via accelerated BMP2-induced Smad1/5/8 phosphorylation

    Science.gov (United States)

    Liu, Jia; Saito, Kan; Maruya, Yuriko; Nakamura, Takashi; Yamada, Aya; Fukumoto, Emiko; Ishikawa, Momoko; Iwamoto, Tsutomu; Miyazaki, Kanako; Yoshizaki, Keigo; Ge, Lihong; Fukumoto, Satoshi

    2016-01-01

    Bone morphogenetic proteins (BMPs) regulate hard tissue formation, including bone and tooth. Growth differentiation factor 5 (GDF5), a known BMP, is expressed in cartilage and regulates chondrogenesis, and mutations have been shown to cause osteoarthritis. Notably, GDF5 is also expressed in periodontal ligament tissue; however, its role during tooth development is unclear. Here, we used cell culture and in vivo analyses to determine the role of GDF5 during tooth development. GDF5 and its associated BMP receptors are expressed at the protein and mRNA levels during postnatal tooth development, particularly at a stage associated with enamel formation. Furthermore, whereas BMP2 was observed to induce evidently the differentiation of enamel-forming ameloblasts, excess GDF5 induce mildly this differentiation. A mouse model harbouring a mutation in GDF5 (W408R) showed enhanced enamel formation in both the incisors and molars, but not in the tooth roots. Overexpression of the W408R GDF5 mutant protein was shown to induce BMP2-mediated mRNA expression of enamel matrix proteins and downstream phosphorylation of Smad1/5/8. These results suggest that mutant GDF5 enhances ameloblast differentiation via accelerated BMP2-signalling. PMID:27030100

  10. Seizure susceptibility of neuropeptide-Y null mutant mice in amygdala kindling and chemical-induced seizure models.

    Science.gov (United States)

    Shannon, Harlan E; Yang, Lijuan

    2004-01-01

    Neuropeptide Y (NPY) administered exogenously is anticonvulsant, and, NPY null mutant mice are more susceptible to kainate-induced seizures. In order to better understand the potential role of NPY in epileptogenesis, the present studies investigated the development of amygdala kindling, post-kindling seizure thresholds, and anticonvulsant effects of carbamazepine and levetiracetam in 129S6/SvEv NPY(+/+) and NPY(-/-) mice. In addition, susceptibility to pilocarpine- and kainate-induced seizures was compared in NPY(+/+) and (-/-) mice. The rate of amygdala kindling development did not differ in the NPY(-/-) and NPY(+/+) mice either when kindling stimuli were presented once daily for at least 20 days, or, 12 times daily for 2 days. However, during kindling development, the NPY(-/-) mice had higher seizure severity scores and longer afterdischarge durations than the NPY(+/+) mice. Post-kindling, the NPY(-/-) mice had markedly lower afterdischarge thresholds and longer afterdischarge durations than NPY (+/+) mice. Carbamazepine and levetiracetam increased the seizure thresholds of both NPY (-/-) and (+/+) mice. In addition, NPY (-/-) mice had lower thresholds for both kainate- and pilocarpine-induced seizures. The present results in amygdala kindling and chemical seizure models suggest that NPY may play a more prominent role in determining seizure thresholds and severity of seizures than in events leading to epileptogenesis. In addition, a lack of NPY does not appear to confer drug-resistance in that carbamazepine and levetiracetam were anticonvulsant in both wild type (WT) and NPY null mutant mice.

  11. ALS mutant FUS proteins are recruited into stress granules in induced pluripotent stem cell-derived motoneurons

    Directory of Open Access Journals (Sweden)

    Jessica Lenzi

    2015-07-01

    Full Text Available Patient-derived induced pluripotent stem cells (iPSCs provide an opportunity to study human diseases mainly in those cases for which no suitable model systems are available. Here, we have taken advantage of in vitro iPSCs derived from patients affected by amyotrophic lateral sclerosis (ALS and carrying mutations in the RNA-binding protein FUS to study the cellular behavior of the mutant proteins in the appropriate genetic background. Moreover, the ability to differentiate iPSCs into spinal cord neural cells provides an in vitro model mimicking the physiological conditions. iPSCs were derived from FUSR514S and FUSR521C patient fibroblasts, whereas in the case of the severe FUSP525L mutation, in which fibroblasts were not available, a heterozygous and a homozygous iPSC line were raised by TALEN-directed mutagenesis. We show that aberrant localization and recruitment of FUS into stress granules (SGs is a prerogative of the FUS mutant proteins and occurs only upon induction of stress in both undifferentiated iPSCs and spinal cord neural cells. Moreover, we show that the incorporation into SGs is proportional to the amount of cytoplasmic FUS, strongly correlating with the cytoplasmic delocalization phenotype of the different mutants. Therefore, the available iPSCs represent a very powerful system for understanding the correlation between FUS mutations, the molecular mechanisms of SG formation and ALS ethiopathogenesis.

  12. ALS mutant FUS proteins are recruited into stress granules in induced pluripotent stem cell-derived motoneurons

    Science.gov (United States)

    Lenzi, Jessica; De Santis, Riccardo; de Turris, Valeria; Morlando, Mariangela; Laneve, Pietro; Calvo, Andrea; Caliendo, Virginia; Chiò, Adriano; Rosa, Alessandro; Bozzoni, Irene

    2015-01-01

    ABSTRACT Patient-derived induced pluripotent stem cells (iPSCs) provide an opportunity to study human diseases mainly in those cases for which no suitable model systems are available. Here, we have taken advantage of in vitro iPSCs derived from patients affected by amyotrophic lateral sclerosis (ALS) and carrying mutations in the RNA-binding protein FUS to study the cellular behavior of the mutant proteins in the appropriate genetic background. Moreover, the ability to differentiate iPSCs into spinal cord neural cells provides an in vitro model mimicking the physiological conditions. iPSCs were derived from FUSR514S and FUSR521C patient fibroblasts, whereas in the case of the severe FUSP525L mutation, in which fibroblasts were not available, a heterozygous and a homozygous iPSC line were raised by TALEN-directed mutagenesis. We show that aberrant localization and recruitment of FUS into stress granules (SGs) is a prerogative of the FUS mutant proteins and occurs only upon induction of stress in both undifferentiated iPSCs and spinal cord neural cells. Moreover, we show that the incorporation into SGs is proportional to the amount of cytoplasmic FUS, strongly correlating with the cytoplasmic delocalization phenotype of the different mutants. Therefore, the available iPSCs represent a very powerful system for understanding the correlation between FUS mutations, the molecular mechanisms of SG formation and ALS ethiopathogenesis. PMID:26035390

  13. Four kinds of ENU-induced white spot mice and chromosome locations of the mutant genes

    Institute of Scientific and Technical Information of China (English)

    WU Baojin; MAO Huihua; SHAO Yixiang; XUE Zhengfeng; LI Houda

    2003-01-01

    Phenotype-driven is the name for an approach used to study gene functions through mutagenesis, location and cloning of the mutant gene. In this study, 150 male C57BL/6J(B6) mice were treated with ENU and reproduced a total offspring of 3860. Of these descendants, 210 exhibited mutation phenotypes by screening, and more than 10 of them are hereditable. Four kinds of mutant mice, named Wbct, W-1Bao, W-2Bao, and W-3Bao, showed dominant hereditary white spot mutation with partial albinism on their belly, distal limbs and tail terminal. To map these mutant genes, 39 microsatellites, equally distributed on the mouse genome and with difference between B6 and DBA/2J (D2), were selected to scan the genome after discrimination of the white spots in the F2 mice [(B6×D2)×D2]. It is found that, the log odds score (LODS) between W-1Bao and D5Mit168 is 0.56, and the LODS of W-1Bao and D5Mit352 is 4.47. With the gradual application of microsatellites D5Mit290, D5Mit312, D5Mit308 and D5Mit356 that are close to the mutant gene, and the number of F2 mice going up to 537, the mutant W-1Bao is located between D5Mit356 and D5Mit308 on chromosome 5, about 42.19 cM from the centromere. In the same way, W-2Bao and W-3Bao are mapped nearby W-1Bao, and Wbct is located on chromosome 1, about 41.6 cM from the centromere. After searching for the mouse genome database (MGD) and performing a one-by-one study of all genes located on chromosome subregion, it is believed that the kit gene is an excellent candidate for the white spot mutations of W-1Bao, W-2Bao and W-3Bao.

  14. CRISPR/Cas9-Induced Double-Strand Break Repair in Arabidopsis Nonhomologous End-Joining Mutants

    Directory of Open Access Journals (Sweden)

    Hexi Shen

    2017-01-01

    Full Text Available Double-strand breaks (DSBs are one of the most harmful DNA lesions. Cells utilize two main pathways for DSB repair: homologous recombination (HR and nonhomologous end-joining (NHEJ. NHEJ can be subdivided into the KU-dependent classical NHEJ (c-NHEJ and the more error-prone KU-independent backup-NHEJ (b-NHEJ pathways, involving the poly (ADP-ribose polymerases (PARPs. However, in the absence of these factors, cells still seem able to adequately maintain genome integrity, suggesting the presence of other b-NHEJ repair factors or pathways independent from KU and PARPs. The outcome of DSB repair by NHEJ pathways can be investigated by using artificial sequence-specific nucleases such as CRISPR/Cas9 to induce DSBs at a target of interest. Here, we used CRISPR/Cas9 for DSB induction at the Arabidopsis cruciferin 3 (CRU3 and protoporphyrinogen oxidase (PPO genes. DSB repair outcomes via NHEJ were analyzed using footprint analysis in wild-type plants and plants deficient in key factors of c-NHEJ (ku80, b-NHEJ (parp1 parp2, or both (ku80 parp1 parp2. We found that larger deletions of >20 bp predominated after DSB repair in ku80 and ku80 parp1 parp2 mutants, corroborating with a role of KU in preventing DSB end resection. Deletion lengths did not significantly differ between ku80 and ku80 parp1 parp2 mutants, suggesting that a KU- and PARP-independent b-NHEJ mechanism becomes active in these mutants. Furthermore, microhomologies and templated insertions were observed at the repair junctions in the wild type and all mutants. Since these characteristics are hallmarks of polymerase θ-mediated DSB repair, we suggest a possible role for this recently discovered polymerase in DSB repair in plants.

  15. CRISPR/Cas9-Induced Double-Strand Break Repair in Arabidopsis Nonhomologous End-Joining Mutants.

    Science.gov (United States)

    Shen, Hexi; Strunks, Gary D; Klemann, Bart J P M; Hooykaas, Paul J J; de Pater, Sylvia

    2017-01-05

    Double-strand breaks (DSBs) are one of the most harmful DNA lesions. Cells utilize two main pathways for DSB repair: homologous recombination (HR) and nonhomologous end-joining (NHEJ). NHEJ can be subdivided into the KU-dependent classical NHEJ (c-NHEJ) and the more error-prone KU-independent backup-NHEJ (b-NHEJ) pathways, involving the poly (ADP-ribose) polymerases (PARPs). However, in the absence of these factors, cells still seem able to adequately maintain genome integrity, suggesting the presence of other b-NHEJ repair factors or pathways independent from KU and PARPs. The outcome of DSB repair by NHEJ pathways can be investigated by using artificial sequence-specific nucleases such as CRISPR/Cas9 to induce DSBs at a target of interest. Here, we used CRISPR/Cas9 for DSB induction at the Arabidopsis cruciferin 3 (CRU3) and protoporphyrinogen oxidase (PPO) genes. DSB repair outcomes via NHEJ were analyzed using footprint analysis in wild-type plants and plants deficient in key factors of c-NHEJ (ku80), b-NHEJ (parp1 parp2), or both (ku80 parp1 parp2). We found that larger deletions of >20 bp predominated after DSB repair in ku80 and ku80 parp1 parp2 mutants, corroborating with a role of KU in preventing DSB end resection. Deletion lengths did not significantly differ between ku80 and ku80 parp1 parp2 mutants, suggesting that a KU- and PARP-independent b-NHEJ mechanism becomes active in these mutants. Furthermore, microhomologies and templated insertions were observed at the repair junctions in the wild type and all mutants. Since these characteristics are hallmarks of polymerase θ-mediated DSB repair, we suggest a possible role for this recently discovered polymerase in DSB repair in plants. Copyright © 2017 Shen et al.

  16. Genetic Analysis and Fine Mapping of a Premature Leaf Senescence Mutant in Rice (Orzya sativa L.)%水稻早衰叶突变体 PLS2的遗传分析与基因定位

    Institute of Scientific and Technical Information of China (English)

    张涛; 郑家奎; 孙玉莹; 郑建敏; 程治军; 蒋开锋; 杨莉; 曹应江; 游书梅; 万建民

    2014-01-01

    叶片早衰引起叶绿素和其他大分子被降解,叶片光合能力降低。这个过程常伴随着活性氧(ROS)的积累,以及细胞中抗氧化酶(SOD、CAT 和 APX)活性的降低,衰老相关基因(SAG)表达量上调,最终导致整个植株过早成熟,产量降低。因此,研究水稻早衰遗传机制和基因功能对于水稻的遗传改良具有重要的作用和意义。PLS2是通过航天育种工程经空间辐射诱变得来的突变体,在孕穗期表现早衰。与野生型相比, PLS2的光合能力降低,株高变矮,节间和穗长缩短,分蘖数和有效分蘖数减少,穗粒数和结实率明显下降,千粒重降低,穗发育不良,灌浆不充分;叶片的CAT 活性显著降低、H2O2积累、死亡细胞增加,叶绿体结构变差,叶绿体中淀粉和嗜锇颗粒增多。黑暗处理加速突变体叶片衰老,叶绿体超微结构球状化。利用 PLS2/蜀恢527和 PLS2/02428的隐性定位群体,将 pls2定位在第3染色体标记 RM14704(8674283 bp)与 SL-I-5(8758394 bp)之间,物理距离84.11 kb,区间内包括14个基因,测序发现在 LOC_Os03g15840第9个外显子第41位的 C 被替换为 T,导致精氨酸(R)替换为半胱氨酸(C), LOC_Os3g15840编码水稻中的一个糖基转移酶(glycosyltransferases, GTs),可能是 pls2的候选基因。为下一步调控基因的克隆和功能研究奠定了基础。%Leaf senescence induces degradation of chlorophyll and other macromolecules, reducing leaf photosynthetic capacity. This process is accompanied by the accumulation of reactive oxygen species (ROS), the decreasing of cell antioxidant enzyme (SOD, CAT, and APX) activity, and the increasing of aging related gene (SAG) expression, leading in early maturity and yield reduction. Therefore, studies on the genetic mechanism and gene function of premature senescence in rice, has the important ef-fect and significance in genetic improvement of rice. PLS2 from space radiation mutation breeding project showed leaf

  17. Genetic Analysis and Gene Mapping of White Stripe Leaf Mutant st11 in Rice%水稻白条纹叶突变体 st11的遗传分析与基因定位

    Institute of Scientific and Technical Information of China (English)

    成钦淑; 李仕贵; 陈学伟; 叶邦全#; 袁灿; 李伟滔; 尹俊杰; 王静; 贺闽; 汪吉春; 王玉平

    2015-01-01

    白条纹叶突变体 st11是从粳稻品种 Kitaake 组培过程中获得的。该突变体在分蘖前叶色表现为正常,从分蘖期开始新生叶表现为白条纹直至成熟期。与野生型相比,该突变体的分蘖、株高、结实率和千粒重等农艺性状没有发生明显变化。遗传分析表明该突变体白条纹叶性状受一对隐性核基因控制。利用该突变体分别与水稻02428、Jodan 杂交构建了两个F2群体用于基因定位。通过集群分离分析(bulked segregant analysis)发现该基因位于第1染色体端粒附近,并与分子标记RM151和 RM10080连锁。进一步利用更多分子标记分析 F2群体,我们将该基因定位于 I10和 I26两个标记之间大约270kb的区间内。%A white stripe leaf mutant st1 1 was obtained from tissue-cultured rice Kitaake (Oryza sativa spp j aponica ). This mutant was characterized by white stripe leaves from tillering stage to mature stage.Compared with the wild type Kitaake,the mutant showed no obvious changes in agricultural traits,such as tiller number,plant height,seed-setting rate,and grain weight.Genetic analyses showed that the white stripe leaf phenotype in the mutant st1 1 was controlled by a single recessive gene.Two F2 segregating populations derived from the crosses,st1 1 ×Jodan and st1 1 ×02428, respectively,were used for mapping the gene st1 1 .Bulked segregant analysis suggested that the gene st1 1 was located close to the markers,RM1 5 1 and RM10080,on chromosome 1 .Using more molecular markers,we finally delimited the gene st1 1 to a region of about 270 kb between the Indel markers,I10 and I26,on the telomere of the short arm of chromosome 1 .

  18. Jasmonoyl-l-isoleucine is required for the production of a flavonoid phytoalexin but not diterpenoid phytoalexins in ultraviolet-irradiated rice leaves.

    Science.gov (United States)

    Miyamoto, Koji; Enda, Isami; Okada, Toshiki; Sato, Yumiko; Watanabe, Kohei; Sakazawa, Tomoko; Yumoto, Emi; Shibata, Kyomi; Asahina, Masashi; Iino, Moritoshi; Yokota, Takao; Okada, Kazunori; Yamane, Hisakazu

    2016-10-01

    Rice produces low-molecular-weight antimicrobial compounds known as phytoalexins, in response to not only pathogen attack but also abiotic stresses including ultraviolet (UV) irradiation. Rice phytoalexins are composed of diterpenoids and a flavonoid. Recent studies have indicated that endogenous jasmonyl-l-isoleucine (JA-Ile) is not necessarily required for the production of diterpenoid phytoalexins in blast-infected or CuCl2-treated rice leaves. However, JA-Ile is required for the accumulation of the flavonoid phytoalexin, sakuranetin. Here, we investigated the roles of JA-Ile in UV-induced phytoalexin production. We showed that UV-irradiation induces the biosynthesis of JA-Ile and its precursor jasmonic acid. We also showed that rice jasmonate biosynthesis mutants produced diterpenoid phytoalexins but not sakuranetin in response to UV, indicating that JA-Ile is required for the production of sakuranetin but not diterpenoid phytoalexins in UV-irradiated rice leaves.

  19. The Phenotype of a Dwarf Mutant in Indica Rice Tm049 and Its Responses to Gibberellin%籼稻矮杆突变体Tm049的表型分析及其对外源赤霉素的响应

    Institute of Scientific and Technical Information of China (English)

    黄秀芝; 丁海东; 梁建生

    2012-01-01

    Abstract: A rice dwarf mutant Tm049 from Indica rice 9311 was obtained in the progeny after 60Co-rays inducement, and the phenotype, internode cell structure and physiological characteristics of Tm049 were investigated. The results showed that Tm049 exhibited dwarf, little dark green and short and wide leaves, fewer tillers, spike erect, small and round grains, and belonging to the dn-type dwarf. The volume and number of cellsin 1st and 2nd internodes were changed. The sensitivity of the second leaf sheaths Tm049 to exogenous gibberellin (GA3) was reduced. The endogenous GA3 levels of Tm049 was slightly higher than that of wild--type 9311. It could be speculated that the mutant may be related to the deletion of the GA signaling pathway. These results will lay a foundation for positional cloning the dwarf gene, functional analysis and molecular marker- assisted selection of new varieties.%利用60Coγ射线辐照籼稻93-11,获得一个性状稳定的矮化突变体材料Tm049,对其表型、节间细胞结构及生理特性方面进行了分析。研究结果表明,Tm049植株矮化、叶色略深且短而宽、分蘖少、穗型直立状、籽粒小而圆、属于dn型矮杆;第一节间和第二节间细胞体积和数目发生变化;第二叶鞘对外源赤霉素(GA3)敏感性降低;内源GA3水平略高于野生型.推测该突变体可能与GA的信号途径缺失有关.研究结果为今后该基因的定位克隆、功能分析及分子标记辅助培育理想株型的新品种奠定了研究基础.

  20. Identification and Studies of a Rice Mutant hya-1 with Semi-dwarf and Aneuploid Traits%一份水稻半矮秆非整倍突变体hya-1的鉴定与研究

    Institute of Scientific and Technical Information of China (English)

    郭涛; 刘永柱; 张建国; 陈志强; 王慧

    2012-01-01

    A semi-dwarfing rice mutant hya-1 , genetically unstable in plant height was obtained through space muta-genesis of Texianzhan 13. Compared to the wild type, hya-1 mutant was featured by semi-dwarfness, fewer tillers, slim culms, reduced grain number per panicle, low seed setting rate and low germination rate. Segregation of plant height in selfed progenies of the mutant was observed. Normal plants of selfed progenies, however, showed genetic stability. Plant height of F, hybrid and F2 progenies generated from the cross between hya-1 mutant and Texianzhan 13 did not follow Mendelian inheritance. Furthermore, selfed-progenies of the mutant had abnormal chromosome numbers (e.g. 2n = 22, 23, 25 and 36 instead of 2n- 24). In addition, there were some pollen mother cells of hya-1 with 11 bivalents at the diakinesis stage of meiosis. Real-time quantitative PCR analysis of normal and semi-dwarf plants from selfed progenies of the mutant along with Texianzhan 13 showed equal copy number of chromosome in normal and wild type plants while the copy number of chromosome 8 and 11 differed in hya-1 and semi-dwarf plants from the selfed progenies. It suggested that hya-1 mutant is a new aneuploid in which abnormal phenotype and genetic instability are caused by abnormal chromosome number and/or varying copy number of certain chromosomes.%通过空间诱变得到半矮秆突变体hya-1,该突变体半矮秆性状遗传稳定性丧失.与其野生型特籼占13相比,hya-1株高显著降低、分蘖少、茎秆细弱、每穗粒数减少、结实率降低、发芽率降低.单株收获半矮株hya-1自交种子,对半矮生性状进行连续选择,发现该性状无法纯合稳定,自交群体株高长期分离(半矮秆和野生型);分离群体中的野生型个体自交,其株高性状稳定.利用hya-1与特籼占13配制杂交组合,其F1及F2株高表型不符合孟德尔分离规律.hya-1自交收获种子根尖细胞染色体数目出现了 2n=22、23、24、25、36的变

  1. pH Induced Conformational Transitions in the Transforming Growth Factor β-Induced Protein (TGFβIp) Associated Corneal Dystrophy Mutants.

    Science.gov (United States)

    Murugan, Elavazhagan; Venkatraman, Anandalakshmi; Lei, Zhou; Mouvet, Victoria; Rui Yi Lim, Rayne; Muruganantham, Nandhakumar; Goh, Eunice; Swee Lim Peh, Gary; Beuerman, Roger W; Chaurasia, Shyam S; Rajamani, Lakshminarayanan; Mehta, Jodhbir S

    2016-03-31

    Most stromal corneal dystrophies are associated with aggregation and deposition of the mutated transforming growth factor-β induced protein (TGFβIp). The 4(th)_FAS1 domain of TGFβIp harbors ~80% of the mutations that forms amyloidogenic and non-amyloidogenic aggregates. To understand the mechanism of aggregation and the differences between the amyloidogenic and non-amyloidogenic phenotypes, we expressed the 4(th)_FAS1 domains of TGFβIp carrying the mutations R555W (non-amyloidogenic) and H572R (amyloidogenic) along with the wild-type (WT). R555W was more susceptible to acidic pH compared to H572R and displayed varying chemical stabilities with decreasing pH. Thermal denaturation studies at acidic pH showed that while WT did not undergo any conformational transition, the mutants exhibited a clear pH-dependent irreversible conversion from αβ conformation to β-sheet oligomers. The β-oligomers of both mutants were stable at physiological temperature and pH. Electron microscopy and dynamic light scattering studies showed that β-oligomers of H572R were larger compared to R555W. The β-oligomers of both mutants were cytotoxic to primary human corneal stromal fibroblast (pHCSF) cells. The β-oligomers of both mutants exhibit variations in their morphologies, sizes, thermal and chemical stabilities, aggregation patterns and cytotoxicities.

  2. Induced Pib Expression and Resistance to Magnaporthe grisea are Compromised by Cytosine Demethylation at Critical Promoter Regions in Rice.

    Science.gov (United States)

    Li, Yuan; Xia, Qiong; Kou, Hongping; Wang, Dan; Lin, Xiuyun; Wu, Ying; Xu, Chunming; Xing, Shaochen; Liu, Bao

    2011-10-01

    Pib is a well-characterized rice blast-resistance gene belonging to the nucleotide binding site (NBS) and leucine-rich repeat (LRR) superfamily. Expression of Pib was low under non-challenged conditions, but strongly induced by the blast-causing fungal pathogen Magnaporthe grisea, thereby conferring resistance to the pathogen. It is generally established that cytosine methylation of the promoter-region often plays a repressive role in modulating expression of the gene in question. We report here that two critical regions of the Pib promoter were heavily CG cytosine-methylated in both cultivars studied. Surprisingly, induced expression of Pib by M. grisea infection did not entail its promoter demethylation, and partial demethylation by 5-azacytidine-treatment actually reduced Pib expression relative to wild-type plants. Accordingly, the blast disease-resistance was compromised in the 5'-azaC-treated plants relative to wild-type. In contrast, the disease susceptibility was not affected by the 5'-azaC treatment in another two rice cultivars that did not contain the Pib gene, ruling out effects of other R genes and non-specific genotoxic effects by the drug-treatment as a cause for the compromised Pib-conditioned blast-resistance. Taken together, our results suggest that promoter DNA methylation plays a novel enhancing role in conditioning high-level of induced expression of the Pib gene in times of M. grisea infection, and its conferred resistance to the pathogen.

  3. Comparative transcriptome analysis of rice seedlings induced by different doses of heavy ion radiation

    Science.gov (United States)

    Zhao, Qian; Sun, Yeqing; Wang, Wei

    2016-07-01

    Highly ionizing radiation (HZE) in space is considered as a main factor causing biological effects on plant seeds. To investigate the different effects on genome-wide gene expression of low-dose and high-dose ion radiation, we carried out ground-base carbon particle HZE experiments with different cumulative doses (0Gy, 0.2Gy, 2Gy) to rice seeds and then performed comparative transcriptome analysis of the rice seedlings. We identified a total of 2551 and 1464 differentially expressed genes (DEGs) in low-dose and high-dose radiation groups, respectively. Gene ontology analyses indicated that low-dose and high-dose ion radiation both led to multiple physiological and biochemical activities changes in rice. By Gene Ontology analyses, the results showed that only one process-oxidation reduction process was enriched in the biological process category after high-dose ion radiation, while more processes such as response to biotic stimulus, heme binding, tetrapyrrole binding, oxidoreductase activity, catalytic activity and oxidoreductase activity were significantly enriched after low-dose ion radiation. The results indicated that the rice plants only focused on the process of oxidation reduction to response to high-dose ion radiation, whereas it was a coordination of multiple biological processes to response to low-dose ion radiation. To elucidate the transcriptional regulation of radiation stress-responsive genes, we identified several DEGs-encoding TFs. AP2/EREBP, bHLH, C2H2, MYB and WRKY TF families were altered significantly in response to ion radiation. Mapman analysis speculated that the biological effects on rice seedlings caused by the radiation stress might share similar mechanisms with the biotic stress. Our findings highlight important alterations in the expression of radiation response genes, metabolic pathways, and TF-encoding genes in rice seedlings exposed to low-dose and high-dose ion radiation.

  4. Isocitrate dehydrogenase 1 mutant R132H sensitizes glioma cells to BCNU-induced oxidative stress and cell death.

    Science.gov (United States)

    Mohrenz, Isabelle Vanessa; Antonietti, Patrick; Pusch, Stefan; Capper, David; Balss, Jörg; Voigt, Sophia; Weissert, Susanne; Mukrowsky, Alicia; Frank, Jan; Senft, Christian; Seifert, Volker; von Deimling, Andreas; Kögel, Donat

    2013-11-01

    Isocitrate dehydrogenase 1 (IDH1) decarboxylates isocitrate to α-ketoglutarate (α-KG) leading to generation of NADPH, which is required to regenerate reduced glutathione (GSH), the major cellular ROS scavenger. Mutation of R132 of IDH1 abrogates generation of α-KG and leads to conversion of α-KG to 2-hydroxyglutarate. We hypothesized that glioma cells expressing mutant IDH1 have a diminished antioxidative capacity and therefore may encounter an ensuing loss of cytoprotection under conditions of oxidative stress. Our study was performed with LN229 cells stably overexpressing IDH1 R132H and wild type IDH1 or with a lentiviral IDH1 knockdown. Quantification of GSH under basal conditions and following treatment with the glutathione reductase inhibitor BCNU revealed significantly lower GSH levels in IDH1 R132H expressing cells and IDH1 KD cells compared to their respective controls. FACS analysis of cell death and ROS production also demonstrated an increased sensitivity of IDH1-R132H-expressing cells and IDH1 KD cells to BCNU, but not to temozolomide. The sensitivity of IDH1-R132H-expressing cells and IDH1 KD cells to ROS induction and cell death was further enhanced with the transaminase inhibitor aminooxyacetic acid and under glutamine free conditions, indicating that these cells were more addicted to glutaminolysis. Increased sensitivity to BCNU-induced ROS production and cell death was confirmed in HEK293 cells inducibly expressing the IDH1 mutants R132H, R132C and R132L. Based on these findings we propose that in addition to its established pro-tumorigenic effects, mutant IDH1 may also limit the resistance of gliomas to specific death stimuli, therefore opening new perspectives for therapy.

  5. Long-day photoperiod induced unhealthy development of chloroplasts in the photoperiod-sensitive genie male-sterile rice

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    By measurement of photochemical activities of chloroplasts and observation on supramolecular archi tecture of thylakoids in chloroplasts, it was found that compared with the effects of short-day photoperiod, long-day pho toperiod could induce normal development of chloroplasts in seedlings of NK58S (photoperiod-sensitive genie male-sterile rice) and NK58 (original line) which do not enter the photoperiod sensitive phase and in seedlings of NK58 just enter the photoperiod-sensitive phase. However, it could induce unhealthy development of chloroplasts in seedlings of NK58S which also just enter the photoperiod sensitive phase. This special effect of long-day photoperiod on the development of chloroplasts in NK58S is probably one of main reasons why long-day photoperiod induces rale-sterility in NK58S and normal fertility in NK58.

  6. Zinc-dependent protection of tobacco and rice cells from Aluminum-induced superoxide-mediated cytotoxicity

    Directory of Open Access Journals (Sweden)

    Cun eLin

    2015-12-01

    Full Text Available Al3+ toxicity in growing plants is considered as one of the major factors limiting the production of crops on acidic soils worldwide. In the last 15 years, it has been proposed that Al3+ toxicity are mediated with distortion of the cellular signaling mechanisms such as calcium signaling pathways, and production of cytotoxic reactive oxygen species (ROS causing oxidative damages. On the other hand, zinc is normally present in plants at high concentrations and its deficiency is one of the most widespread micronutrient deficiencies in plants. Earlier studies suggested that lack of zinc often results in ROS-mediated oxidative damage to plant cells. Previously, inhibitory action of Zn2+ against lanthanide-induced superoxide generation in tobacco cells have been reported, suggesting that Zn2+ interferes with the cation-induced ROS production via stimulation of NADPH oxidase. In the present study, the effect of Zn2+ on Al3+-induced superoxide generation in the cell suspension cultures of tobacco (Nicotiana tabacum L., cell-line, BY-2 and rice (Oryza sativa L., cv. Nipponbare, was examined. The Zn2+-dependent inhibition of the Al3+-induced oxidative burst was observed in both model cells selected from the monocots and dicots (rice and tobacco, suggesting that this phenomenon (Al3+/Zn2+ interaction can be preserved in higher plants. Subsequently induced cell death in tobacco cells was analyzed by lethal cell staining with Evans blue. Obtained results indicated that presence of Zn2+ at physiological concentrations can protect the cells by preventing the Al3+-induced superoxide generation and cell death. Furthermore, the regulation of the Ca2+ signaling, i.e. change in the cytosolic Ca2+ ion concentration, and the cross-talks among the elements which participate in the pathway were further explored.

  7. Elucidation of complex nature of PEG induced drought-stress response in rice root using comparative proteomics approach

    Directory of Open Access Journals (Sweden)

    Lalit Agrawal

    2016-09-01

    Full Text Available Along with many adaptive strategies, dynamic changes in protein abundance seem to be the common strategy to cope up with abiotic stresses which can be best explored through proteomics. Understanding of drought response is the key to decipher regulatory mechanism of better adaptation. Rice (Oryza sativa L. proteome represents a phenomenal source of proteins that govern traits of agronomic importance, such as drought tolerance. In this study, a comparison of root cytoplasmic proteome was done for a drought tolerant rice (Heena cultivar in PEG induced drought conditions. A total of 510 protein spots were observed by PDQuest analysis and 125 differentially regulated spots were subjected for MALDI-TOF MS-MS analysis out of which 102 protein spots identified which further led to identification of 78 proteins with a significant score. These 78 differentially expressed proteins appeared to be involved in different biological pathways. The largest percentage of identified proteins was involved in bioenergy and metabolism (29% and mainly consists of malate dehydrogenase, succinyl-CoA, putative acetyl-CoA synthetase and pyruvate dehydrogenase etc. This was followed by proteins related to cell defense and rescue (22% such as monodehydroascorbate reductase and stress-induced protein sti1, then by protein biogenesis and storage class (21% e.g. putative thiamine biosynthesis protein, putative beta-alanine synthase and cysteine synthase. Further, cell signaling (9% proteins like actin and prolyl endopeptidase and proteins with miscellaneous function (19% like Sgt1 and some hypothetical protein were also represented a large contribution towards drought regulatory mechanism in rice. We propose that protein biogenesis, cell defense and superior homeostasis may render better drought-adaptation. These findings might expedite the functional determination of the drought-responsive proteins and their prioritisation as potential molecular targets for perfect adaptation.

  8. Elucidation of Complex Nature of PEG Induced Drought-Stress Response in Rice Root Using Comparative Proteomics Approach

    Science.gov (United States)

    Agrawal, Lalit; Gupta, Swati; Mishra, Shashank K.; Pandey, Garima; Kumar, Susheel; Chauhan, Puneet S.; Chakrabarty, Debasis; Nautiyal, Chandra S.

    2016-01-01

    Along with many adaptive strategies, dynamic changes in protein abundance seem to be the common strategy to cope up with abiotic stresses which can be best explored through proteomics. Understanding of drought response is the key to decipher regulatory mechanism of better adaptation. Rice (Oryza sativa L.) proteome represents a phenomenal source of proteins that govern traits of agronomic importance, such as drought tolerance. In this study, a comparison of root cytoplasmic proteome was done for a drought tolerant rice (Heena) cultivar in PEG induced drought conditions. A total of 510 protein spots were observed by PDQuest analysis and 125 differentially regulated spots were subjected for MALDI-TOF MS-MS analysis out of which 102 protein spots identified which further led to identification of 78 proteins with a significant score. These 78 differentially expressed proteins appeared to be involved in different biological pathways. The largest percentage of identified proteins was involved in bioenergy and metabolism (29%) and mainly consists of malate dehydrogenase, succinyl-CoA, putative acetyl-CoA synthetase, and pyruvate dehydrogenase etc. This was followed by proteins related to cell defense and rescue (22%) such as monodehydroascorbate reductase and stress-induced protein sti1, then by protein biogenesis and storage class (21%) e.g. putative thiamine biosynthesis protein, putative beta-alanine synthase, and cysteine synthase. Further, cell signaling (9%) proteins like actin and prolyl endopeptidase, and proteins with miscellaneous function (19%) like Sgt1 and some hypothetical proteins were also represented a large contribution toward drought regulatory mechanism in rice. We propose that protein biogenesis, cell defense, and superior homeostasis may render better drought-adaptation. These findings might expedite the functional determination of the drought-responsive proteins and their prioritization as potential molecular targets for perfect adaptation.

  9. Selenium ameliorates arsenic induced oxidative stress through modulation of antioxidant enzymes and thiols in rice (Oryza sativa L.).

    Science.gov (United States)

    Kumar, Amit; Singh, Rana Pratap; Singh, Pradyumna Kumar; Awasthi, Surabhi; Chakrabarty, Debasis; Trivedi, Prabodh Kumar; Tripathi, Rudra Deo

    2014-09-01

    Arsenic (As) contamination of rice is a major problem for South-East Asia. In the present study, the effect of selenium (Se) on rice (Oryza sativa L.) plants exposed to As was studied in hydroponic culture. Arsenic accumulation, plant growth, thiolic ligands and antioxidative enzyme activities were assayed after single (As and Se) and simultaneous supplementations (As + Se). The results indicated that the presence of Se (25 µM) decreased As accumulation by threefold in roots and twofold in shoots as compared to single As (25 µM) exposed plants. Arsenic induced oxidative stress in roots and shoots was significantly ameliorated by Se supplementation. The observed positive response was found associated with the increased activities of ascorbate peroxidase (APX; EC 1.11.1.11), catalase (CAT; EC 1.11.1.6) and glutathione peroxidase (GPx; EC 1.11.1.9) and induced levels of non-protein thiols (NPTs), glutathione (GSH) and phytochelatins (PCs) in As + Se exposed plants as compared to single As treatment. Selenium supplementation modulated the thiol metabolism enzymes viz., γ-glutamylcysteine synthetase (γ-ECS; EC 6.3.2.2), glutathione-S-transferase (GST; EC 2.5.1.18) and phytochelatin synthase (PCS; EC 2.3.2.15). Gene expression analysis of several metalloid responsive genes (LOX, SOD and MATE) showed upregulation during As stress, however, significant downregulation during As + Se exposure as compared to single As treatment. Gene expressions of enzymes of antioxidant and GSH and PC biosynthetic systems, such as APX, CAT, GPx, γ-ECS and PCS were found to be significantly positively correlated with their enzyme activities. The findings suggested that Se supplementation could be an effective strategy to reduce As accumulation and toxicity in rice plants.

  10. Role of various enterotoxins in Aeromonas hydrophila-induced gastroenteritis: generation of enterotoxin gene-deficient mutants and evaluation of their enterotoxic activity.

    Science.gov (United States)

    Sha, Jian; Kozlova, E V; Chopra, A K

    2002-04-01

    Three enterotoxins from the Aeromonas hydrophila diarrheal isolate SSU have been molecularly characterized in our laboratory. One of these enterotoxins is cytotoxic in nature, whereas the other two are cytotonic enterotoxins, one of them heat labile and the other heat stable. Earlier, by developing an isogenic mutant, we demonstrated the role of a cytotoxic enterotoxin in causing systemic infection in mice. In the present study, we evaluated the role of these three enterotoxins in evoking diarrhea in a murine model by developing various combinations of enterotoxin gene-deficient mutants by marker-exchange mutagenesis. A total of six isogenic mutants were prepared in a cytotoxic enterotoxin gene (act)-positive or -negative background strain of A. hydrophila. We developed two single knockouts with truncation in either the heat-labile (alt) or the heat-stable (ast) cytotonic enterotoxin gene; three double knockouts with truncations of genes encoding (i) alt and ast, (ii) act and alt, and (iii) act and ast genes; and a triple-knockout mutant with truncation in all three genes, act, alt, and ast. The identity of these isogenic mutants developed by double-crossover homologous recombination was confirmed by Southern blot analysis. Northern and Western blot analyses revealed that the expression of different enterotoxin genes in the mutants was correspondingly abrogated. We tested the biological activity of these mutants in a diet-restricted and antibiotic-treated mouse model with a ligated ileal loop assay. Our data indicated that all of these mutants had significantly reduced capacity to evoke fluid secretion compared to that of wild-type A. hydrophila; the triple-knockout mutant failed to induce any detectable level of fluid secretion. The biological activity of selected A. hydrophila mutants was restored after complementation. Taken together, we have established a role for three enterotoxins in A. hydrophila-induced gastroenteritis in a mouse model with the greatest

  11. Padrão de integração de pAN7-1 em mutantes de Magnaporthe grisea com patogenicidade alterada em arroz pAN7-1 integration pattern in mutants of Magnaporthe grisea impaired in pathogenicity to rice

    Directory of Open Access Journals (Sweden)

    Carlos Eduardo Marchi

    2010-03-01

    Full Text Available Ensaios foram conduzidos para verificar a presença, o número de cópias e de sítios de integração de pAN7-1 no genoma de mutantes de M. grisea I-22 com patogenicidade alterada em arroz. Foram analisados T41, T93, T251 (gerados por mutagênese REMI e T108 (oriundo de mutagênese convencional, os quais exibiram diferentes fenótipos mutantes. O DNA total desses mutantes foi submetido à reação em cadeia de polimerase (PCR e às análises de hibridização com o vetor (Southern blot. A presença de pAN7-1 no genoma de todos os mutantes foi confirmada por PCR. Segundo as análises de Southern blot, T41 exibiu duas integrações do vetor, ambas na forma de cópia única. No genoma de T93 também foram detectados dois sítios de inserção de pAN7-1, um dos quais envolvendo múltiplas cópias do vetor. Os resultados indicaram a presença de apenas uma cópia do vetor em um único sítio nos genomas de T108 e T251. O padrão de integração em T251 foi o único a sugerir a ocorrência de evento REMI. As diferenças quanto ao tamanho dos fragmentos com homologia a pAN7-1 refletiram a possível aleatoriedade dos eventos de integração no genoma de M. grisea. Os resultados evidenciaram o potencial de REMI para a mutagênese insercional de M. grisea, quando conduzida com pAN7-1 e HindIIIExperiments were conducted to investigate the presence, number of copies and integration site of pAN7-1 in the genome of mutants of Magnaporthe grisea impaired in pathogenicity to rice. The mutants T41, T93, T251 (obtained by REMI and T108 (obtained by conventional mutagenesis exhibiting different alterations in pathogenicity were analyzed. Polymerase chain reaction (PCR and Southern blot analyses were conducted with total DNA of mutants. The presence of pAN7-1 in genome of mutants was confirmed by PCR. Based in Southern blot analyses, two single-copy integration of vector were detected in T41. Two insertion sites of pAN7-1 were detected in the genome of T93

  12. RNAseq analysis reveals pathways and candidate genes associated with salinity tolerance in a spaceflight-induced wheat mutant.

    Science.gov (United States)

    Xiong, Hongchun; Guo, Huijun; Xie, Yongdun; Zhao, Linshu; Gu, Jiayu; Zhao, Shirong; Li, Junhui; Liu, Luxiang

    2017-06-02

    Salinity stress has become an increasing threat to food security worldwide and elucidation of the mechanism for salinity tolerance is of great significance. Induced mutation, especially spaceflight mutagenesis, is one important method for crop breeding. In this study, we show that a spaceflight-induced wheat mutant, named salinity tolerance 1 (st1), is a salinity-tolerant line. We report the characteristics of transcriptomic sequence variation induced by spaceflight, and show that mutations in genes associated with sodium ion transport may directly contribute to salinity tolerance in st1. Furthermore, GO and KEGG enrichment analysis of differentially expressed genes (DEGs) between salinity-treated st1 and wild type suggested that the homeostasis of oxidation-reduction process is important for salt tolerance in st1. Through KEGG pathway analysis, "Butanoate metabolism" was identified as a new pathway for salinity responses. Additionally, key genes for salinity tolerance, such as genes encoding arginine decarboxylase, polyamine oxidase, hormones-related, were not only salt-induced in st1 but also showed higher expression in salt-treated st1 compared with salt-treated WT, indicating that these genes may play important roles in salinity tolerance in st1. This study presents valuable genetic resources for studies on transcriptome variation caused by induced mutation and the identification of salt tolerance genes in crops.

  13. Cloning and characterization of rice RH3 gene induced by brown planthopper

    Institute of Scientific and Technical Information of China (English)

    WANG Xiaolan; WENG Qingmei; YOU Aiqing; ZHU Lili; HE Guangcun

    2003-01-01

    Experiments have showed that the histone H3 gene is correlated with development, cell speciality and stress response. The RH3 full-length cDNA was isolated from the cDNA library of rice infested by brown planthopper (BPH) with EST (Accession no. BU572343) screened from rice SSH library as probe. This gene encodes histone H3 protein including 136 amino acids, with one amino acid different from a kind of disease resistance-related protein in rice (AF467728). At the position 126, the aspartic acid is replaced by lysine. The time course results showed that the expression of the RH3 began to increase at 8 h after BPH-feeding, and got to its peak at 96 h. Regulations of the gene expression in treatments with stress/defense signal molecules were analyzed by Northern blot. Water deficit and Pyricularia grisea increased the expression of RH3 while ABA down-regulated the gene. The enhanced accumulation of RH3 transcripts in the vascular bundle and short cell of stem after BPH feeding was revealed by RNA in situ hybridization. It is the first time to report that RH3 is correlated with the response of rice to BPH.

  14. Inhibiting efficacy of metabolites of Streptomyces lavendulohygtroscopicus and its ultraviolet induced strain on two rice diseases

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    @@ More than 70 Streptomyces lavendulohygtroscopicus strains were isolated from the soil samples collected under different ecological conditions. Fermentation centrifugal supernatant (FCS) of the strians were used as biological control agents to bioassay rice disease pathogens of sheath blight (R. solani) and bakanae (G. fujikuroi).

  15. FT-like proteins induce transposon silencing in the shoot apex during floral induction in rice.

    Science.gov (United States)

    Tamaki, Shojiro; Tsuji, Hiroyuki; Matsumoto, Ayana; Fujita, Akiko; Shimatani, Zenpei; Terada, Rie; Sakamoto, Tomoaki; Kurata, Tetsuya; Shimamoto, Ko

    2015-02-24

    Floral induction is a crucial developmental step in higher plants. Florigen, a mobile floral activator that is synthesized in the leaf and transported to the shoot apex, was recently identified as a protein encoded by FLOWERING LOCUS T (FT) and its orthologs; the rice florigen is Heading date 3a (Hd3a) protein. The 14-3-3 proteins mediate the interaction of Hd3a with the transcription factor OsFD1 to form a ternary structure called the florigen activation complex on the promoter of OsMADS15, a rice APETALA1 ortholog. However, crucial information, including the spatiotemporal overlap among FT-like proteins and the components of florigen activation complex and downstream genes, remains unclear. Here, we confirm that Hd3a coexists, in the same regions of the rice shoot apex, with the other components of the florigen activation complex and its transcriptional targets. Unexpectedly, however, RNA-sequencing analysis of shoot apex from wild-type and RNA-interference plants depleted of florigen activity revealed that 4,379 transposable elements (TEs; 58% of all classifiable rice TEs) were expressed collectively in the vegetative and reproductive shoot apex. Furthermore, in the reproductive shoot apex, 214 TEs were silenced by florigen. Our results suggest a link between floral induction and regulation of TEs.

  16. Expression of mutant TDP-43 induces neuronal dysfunction in transgenic mice

    Directory of Open Access Journals (Sweden)

    Dickson Dennis W

    2011-10-01

    Full Text Available Abstract Background Abnormal distribution, modification and aggregation of transactivation response DNA-binding protein 43 (TDP-43 are the hallmarks of multiple neurodegenerative diseases, especially frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD-U and amyotrophic lateral sclerosis (ALS. Researchers have identified 44 mutations in the TARDBP gene that encode TDP-43 as causative for cases of sporadic and familial ALS http://www.molgen.ua.ac.be/FTDMutations/. Certain mutant forms of TDP-43, such as M337V, are associated with increased low molecular weight (LMW fragments compared to wild-type (WT TDP-43 and cause neuronal apoptosis and developmental delay in chick embryos. Such findings support a direct link between altered TDP-43 function and neurodegeneration. Results To explore the pathogenic properties of the M337V mutation, we generated and characterized two mouse lines expressing human TDP-43 (hTDP-43M337V carrying this mutation. hTDP-43M337V was expressed primarily in the nuclei of neurons in the brain and spinal cord, and intranuclear and cytoplasmic phosphorylated TDP-43 aggregates were frequently detected. The levels of TDP-43 LMW products of ~25 kDa and ~35 kDa species were also increased in the transgenic mice. Moreover, overexpression of hTDP-43M337V dramatically down regulated the levels of mouse TDP-43 (mTDP-43 protein and RNA, indicating TDP-43 levels are tightly controlled in mammalian systems. TDP-43M337V mice displayed reactive gliosis, widespread ubiquitination, chromatolysis, gait abnormalities, and early lethality. Abnormal cytoplasmic mitochondrial aggregates and abnormal phosphorylated tau were also detected in the mice. Conclusion Our novel TDP-43M337V mouse model indicates that overexpression of hTDP-43M337V alone is toxic in vivo. Because overexpression of hTDP-43 in wild-type TDP-43 and TDP-43M337V mouse models produces similar phenotypes, the mechanisms causing pathogenesis in the mutant

  17. Identification of a high frequency transposon induced by tissue culture, nDaiZ, a member of the hAT family in rice.

    Science.gov (United States)

    Huang, Jian; Zhang, Kewei; Shen, Yi; Huang, Zejun; Li, Ming; Tang, Ding; Gu, Minghong; Cheng, Zhukuan

    2009-03-01

    Recent completion of rice genome sequencing has revealed that more than 40% of its genome consists of repetitive sequences, and most of them are related to inactive transposable elements. In the present study, a transposable element, nDaiZ0, which is induced by tissue culture with high frequency, was identified by sequence analysis of an allelic line of the golden hull and internode 2 (gh2) mutant, which was integrated into the forth exon of GH2. The 528-bp nDaiZ0 has 14-bp terminal inverted repeats (TIRs), and generates an 8-bp duplication of its target sites (TSD) during its mobilization. nDaiZs are non-autonomous transposons and have no coding capacity. Bioinformatics analysis and southern blot hybridization showed that at least 16 copies of nDaiZ elements exist in the japonica cultivar Nipponbare genome and 11 copies in the indica cultivar 93-11 genome. During tissue culture, only one copy, nDaiZ9, located on chromosome 5 in the genome of Nipponbare can be activated with its transposable frequency reaching 30%. However, nDaiZ9 was not present in the 93-11 genome. The larger elements, DaiZs, were further identified by database searching using nDaiZ0 as a query because they share similar TIRs and subterminal sequences. DaiZ can also generate an 8-bp TSD. DaiZ elements contain a conserved region with a high similarity to the hAT dimerization motif, suggesting that the nDaiZ-DaiZ transposon system probably belongs to the hAT superfamily of class II transposons. Phylogenetic analysis indicated that it is a new type of plant hAT-like transposon. Although nDaiZ is activated by tissue culture, the high transposable frequency indicates that it could become a useful gene tagging system for rice functional genomic studies. In addition, the mechanism of the high transposable ability of nDaiZ9 is discussed.

  18. Lithium Down-regulates Histone Deacetylase 1 (HDAC1) and Induces Degradation of Mutant Huntingtin*

    Science.gov (United States)

    Wu, Shuai; Zheng, Shui-Di; Huang, Hong-Ling; Yan, Li-Chong; Yin, Xiao-Fei; Xu, Hai-Neng; Zhang, Kang-Jian; Gui, Jing-Hua; Chu, Liang; Liu, Xin-Yuan

    2013-01-01

    Lithium is an effective mood stabilizer that has been clinically used to treat bipolar disorder for several decades. Recent studies have suggested that lithium possesses robust neuroprotective and anti-tumor properties. Thus far, a large number of lithium targets have been discovered. Here, we report for the first time that HDAC1 is a target of lithium. Lithium significantly down-regulated HDAC1 at the translational level by targeting HDAC1 mRNA. We also showed that depletion of HDAC1 is essential for the neuroprotective effects of lithium and for the lithium-mediated degradation of mutant huntingtin through the autophagic pathway. Our studies explain the multiple functions of lithium and reveal a novel mechanism for the function of lithium in neurodegeneration. PMID:24165128

  19. Neuroprotective effects of germinated brown rice against hydrogen peroxide induced cell death in human SH-SY5Y cells.

    Science.gov (United States)

    Ismail, Norsharina; Ismail, Maznah; Fathy, Siti Farhana; Musa, Siti Nor Asma; Imam, Mustapha Umar; Foo, Jhi Biau; Iqbal, Shahid

    2012-01-01

    The neuroprotective and antioxidative effects of germinated brown rice (GBR), brown rice (BR) and commercially available γ-aminobutyric acid (GABA) against cell death induced by hydrogen peroxide (H(2)O(2)) in human neuroblastoma SH-SY5Y cells have been investigated. Results show that GBR suppressed H(2)O(2)-mediated cytotoxicity and induced G0/G1 phase cell cycle arrest in SH-SY5Y cells. Moreover, GBR reduced mitochondrial membrane potential (MMP) and prevented phosphatidylserine (PS) translocation in SH-SY5Y cells, key features of apoptosis, and subsequent cell death. GBR exhibited better neuroprotective and antioxidative activities as compared to BR and GABA. These results indicate that GBR possesses high antioxidative activities and suppressed cell death in SH-SY5Y cells by blocking the cell cycle re-entry and apoptotic mechanisms. Therefore, GBR could be developed as a value added functional food to prevent neurodegenerative diseases caused by oxidative stress and apoptosis.

  20. Transcriptomics and knockout mutant analysis of rhizobacteria-mediated induced systemic resistance in Arabidopsis

    NARCIS (Netherlands)

    Verhagen, B.W.M.

    2004-01-01

    A classic example of induced resistance is triggered after infection by a necrotizing pathogen, rendering uninfected,distal parts more resistant to subsequent pathogen attack, and is often referred to as systemic acquired resistance (SAR). A phenotypically comparable type of induced resistance is tr

  1. Huntingtin-associated protein 1 (HAP1) interacts with androgen receptor (AR) and suppresses SBMA-mutant-AR-induced apoptosis.

    Science.gov (United States)

    Takeshita, Yukio; Fujinaga, Ryutaro; Zhao, Changjiu; Yanai, Akie; Shinoda, Koh

    2006-08-01

    Huntingtin-associated protein 1 (HAP1), an interactor of huntingtin, has been known as an essential component of the stigmoid body (STB) and recently reported to play a protective role against neurodegeneration in Huntington's disease (HD). In the present study, subcellular association between HAP1 and androgen receptor (AR) with a long polyglutamine tract (polyQ) derived from spinal-and-bulbar-muscular-atrophy (SBMA) was examined using HEp-2 cells cotransfected with HAP1 and/or normal ARQ25, SBMA-mutant ARQ65 or deletion-mutant AR cDNAs. The results provided the first clear evidence that HAP1 interacts with AR through its ligand-binding domain in a polyQ-length-dependent manner and forms prominent inclusions sequestering polyQ-AR, and that addition of dihydrotestosterone reduces the association strength of HAP1 with ARQ25 more dramatically than that with ARQ65. Furthermore, SBMA-mutant-ARQ65-induced apoptosis was suppressed by cotransfection with HAP1. Our findings strongly suggest that HAP1/STB is relevant to polyQ-length-dependent modification on subcellular AR functions and critically involved in pathogenesis of not only HD but also SBMA as an important intrinsic neuroprotectant determining the threshold for cellular vulnerability to apoptosis. Taking together with previous reports that HAP1/STB is selectively expressed in the brain regions spared from degenerative targets in HD and SBMA, the current study might explain the region-specific occurrence of neurodegeneration in both diseases, shedding light on common aspects of their molecular pathological mechanism and yet-to-be-uncovered diagnostic or therapeutic applications for HD and SBMA patients.

  2. Inhibition of NF-κb by mutant IκBα enhances TNF-α-induced apoptosis in HL-60 cells by controlling bcl-XL expression

    Institute of Scientific and Technical Information of China (English)

    曹文静; 张瑶珍; 张东华; 李登举; 唐锦治

    2004-01-01

    Backgound The aim of this study was to explore whether the inhibition of nuclear factor-κB (NF-κB) activation by mutant IκBα (S32,36→A) can enhance TNF-α-induced apoptosis of leukemia cells and to investigate the possible mechanism.Methods The mutant IκBα gene was transfected into HL-60 cells by liposome-mediated techniques. G418 resistant clones stably expressing mutant IκBα were obtained by the limiting dilution method. TNF-α-induced NF-κB activation was measured by electrophoretic mobility shift assay (EMSA). The expression of bcl-xL was detected by RT-PCR and Western blot after 4 hours exposure of parental HL-60 and transfected HL-60 cells to a variety of concentrations of TNF-α. The percentage of apoptotic leukemia cells was evaluated by flow cytometry (FCM). Results Mutant IκBα protein was confirmed to exist by Western blot. The results of EMSA showed that NF-κB activation by TNF-α in HL-60 cells was induced in a dose-dependent manner, but was almost completely inhibited by mutant IκBα repressor in transfected cells. The levels of bcl-xL mRNA and protein in HL-60 cells increased after exposure to TNF-α, but changed very little in transfected HL-60 cells. The inhibition of NF-κB activation by mutant IκBα enhanced TNF-α-induced apoptosis. The cytotoxic effects of TNF-α were amplified in a time- and dose-dependent manner.Conclusions NF-κB activation plays an important role in the resistance to TNF-α-induced apoptosis. The inhibition of NF-κB by mutant IκBα could provide a new approach that may enhance the anti-leukemia effects of TNF-α or even of other cytotoxic agents.

  3. Tlr4-mutant mice are resistant to acute alcohol-induced sterol-regulatory element binding protein activation and hepatic lipid accumulation

    Science.gov (United States)

    Zhang, Zhi-Hui; Liu, Xiao-Qian; Zhang, Cheng; He, Wei; Wang, Hua; Chen, Yuan-Hua; Liu, Xiao-Jing; Chen, Xi; Xu, De-Xiang

    2016-01-01

    Previous studies demonstrated that acute alcohol intoxication caused hepatic lipid accumulation. The present study showed that acute alcohol intoxication caused hepatic lipid accumulation in Tlr4-wild-type mice but not in Tlr4-mutant mice. Hepatic sterol-regulatory element binding protein (SREBP)-1, a transcription factor regulating fatty acid and triglyceride (TG) synthesis, was activated in alcohol-treated Tlr4-wild-type mice but not in Tlr4-mutant mice. Hepatic Fas, Acc, Scd-1 and Dgat-2, the key genes for fatty acid and TG synthesis, were up-regulated in alcohol-treated Tlr4-wild-type mice but not in Tlr4-mutant mice. Additional experiment showed that hepatic MyD88 was elevated in alcohol-treated Tlr4-wild-type mice but not in Tlr4-mutant mice. Hepatic NF-κB was activated in alcohol-treated Tlr4-wild-type mice but not in Tlr4-mutant mice. Moreover, hepatic GSH content was reduced and hepatic MDA level was elevated in alcohol-treated Tlr4-wild-type mice but not in Tlr4-mutant mice. Hepatic CYP2E1 was elevated in alcohol-treated Tlr4-wild-type mice but not in Tlr4-mutant mice. Hepatic p67phox and gp91phox, two NADPH oxidase subunits, were up-regulated in alcohol-treated Tlr4-wild-type mice but not in Tlr4-mutant mice. Alpha-phenyl-N-t-butylnitrone (PBN), a free radical spin-trapping agent, protected against alcohol-induced hepatic SREBP-1 activation and hepatic lipid accumulation. In conclusion, Tlr4-mutant mice are resistant to acute alcohol-induced hepatic SREBP-1 activation and hepatic lipid accumulation. PMID:27627966

  4. Nullbasic, a potent anti-HIV tat mutant, induces CRM1-dependent disruption of HIV rev trafficking.

    Directory of Open Access Journals (Sweden)

    Min-Hsuan Lin

    Full Text Available Nullbasic, a mutant of the HIV-1 Tat protein, has anti-HIV-1 activity through mechanisms that include inhibition of Rev function and redistribution of the HIV-1 Rev protein from the nucleolus to the nucleoplasm and cytoplasm. Here we investigate the mechanism of this effect for the first time, establishing that redistribution of Rev by Nullbasic is not due to direct interaction between the two proteins. Rather, Nullbasic affects subcellular localization of cellular proteins that regulate Rev trafficking. In particular, Nullbasic induced redistribution of exportin 1 (CRM1, nucleophosmin (B23 and nucleolin (C23 from the nucleolus to the nucleus when Rev was coexpressed, but never in its absence. Inhibition of the Rev:CRM1 interaction by leptomycin B or a non-interacting RevM10 mutant completely blocked redistribution of Rev by Nullbasic. Finally, Nullbasic did not inhibit importin β- or transportin 1-mediated nuclear import, suggesting that cytoplasmic accumulation of Rev was due to increased export by CRM1. Overall, our data support the conclusion that CRM1-dependent subcellular redistribution of Rev from the nucleolus by Nullbasic is not through general perturbation of either nuclear import or export. Rather, Nullbasic appears to interact with and disrupt specific components of a Rev trafficking complex required for its nucleocytoplasmic shuttling and, in particular, its nucleolar accumulation.

  5. The use of the ph1b mutant to induce recombination between the chromosomes of wheat and barley

    Science.gov (United States)

    Rey, María-Dolores; Calderón, María C.; Prieto, Pilar

    2015-01-01

    Intensive breeding has led to a narrowing in the genetic base of our major crops. In wheat, access to the extensive gene pool residing in its many and varied relatives (some cultivated, others wild) is hampered by the block on recombination imposed by the Ph1 (Pairing homoeologous 1) gene. Here, the ph1b mutant has been exploited to induced allosyndesis between wheat chromosomes and those of both Hordeum vulgare (cultivated barley) and H. chilense (a wild barley). A number of single chromosome Hordeum sp. substitution and addition lines in wheat were crossed and backcrossed to the ph1b mutant to produce plants in which pairing between the wheat and the non-wheat chromosomes was not suppressed by the presence of Ph1. Genomic in situ hybridization was applied to almost 500 BC1F2 progeny as a screen for allosyndetic recombinants. Chromosome rearrangements were detected affecting H. chilense chromosomes 4Hch, 5Hch, 6Hch, and 7Hch and H. vulgare chromosomes 4Hv, 6Hv, and 7Hv. Two of these were clearly the product of a recombination event involving chromosome 4Hch and a wheat chromosome. PMID:25852713

  6. Identification of Francisella novicida mutants that fail to induce prostaglandin E2 synthesis by infected macrophages

    OpenAIRE

    Woolard, Matthew D.; Barrigan, Lydia M.; Fuller, James R.; Buntzman, Adam S.; Bryan, Joshua; Manoil, Colin; Kawula, Thomas H.; Frelinger, Jeffrey A.

    2013-01-01

    Francisella tularensis is the causative agent of tularemia. We have previously shown that infection with F. tularensis Live Vaccine Strain (LVS) induces macrophages to synthesize prostaglandin E2 (PGE2). Synthesis of PGE2 by F. tularensis infected macrophages results in decreased T cell proliferation in vitro and increased bacterial survival in vivo. Although we understand some of the biological consequences of F. tularensis induced PGE2 synthesis by macrophages, we do not understand the cell...

  7. Identification of Francisella novicida mutants that fail to induce prostaglandin E2 synthesis by infected macrophages.

    OpenAIRE

    Matthew Dale Woolard; Barrigan, Lydia M.; Fuller, James R.; Buntzman, Adam S.; Joshua eBryan; Colin eManoil; Tom eKawula; Frelinger, Jeffrey A.

    2013-01-01

    Francisella tularensis is the causative agent of tularemia. We have previously shown that infection with F. tularensis Live Vaccine Strain (LVS) induces macrophages to synthesize prostaglandin E2 (PGE2). Synthesis of PGE2 by F. tularensis infected macrophages results in decreased T cell proliferation in vitro and increased bacterial survival in vivo. Although we understand some of the biological consequences of F. tularensis induced PGE2 synthesis by macrophages, we do not understand the ce...

  8. Xanthomonas oryzae pv oryzae the Causal Agent of Bacterial Leaf Blight of rice: Isolation, Characterization, and Study of Transposon Mutagenesis

    Directory of Open Access Journals (Sweden)

    Abdjad Asih Nawangsih

    2011-04-01

    Full Text Available Xanthomonas oryzae pv oryzae the Causal Agent of Bacterial Leaf Blight of rice: Isolation, Characterization, and Study of Transposon Mutagenesis. X. oryzae pv. oryzae (Xoo causes bacterial leaf blight (BLB of rice (Oryza sativa L., a major disease that constrains production of the staple crop in many countries of the world. Identification of X. oryzae pv. oryzae (Xoo was conducted based on the disease symptoms, pathogenicity, morphological, physiological, and genetic characteristics of bacterial cultures isolated from the infected plants. Fifty bacterial isolates predicted as Xoo have been successfully isolated. They are aerobic, rod shaped, and Gram negative bacteria. The isolates were evaluated for their hypersensitivity in tobacco and pathogenicity in rice plant. Fifty isolates induced hypersensitive reaction in tobacco and showed pathogenicity symptom in rice in different length. Based on physiological test, hypersensitivity and pathogenicity reactions, three bacterial isolates strongly predicted as Xoo, i.e. STG21, STG42, and STG46, were non indole formation, non pigment fluorescent, hydrolyzed casein, catalase activity positive, but negative oxidase. Partial sequencing of 16S rRNA genes of STG21 and STG42 showed 80% and 82% homology with X. oryzae, respectively, while STG46 showed 84% homology with X. campestris. Mini-Tn5 transposon mutagenesis of STG21 generated one of the mutants (M5 lossed it’s ability to induce hypersensitive reaction in tobacco plant and deficient in pathogenicity on rice. The lesion length of rice leaf caused by the mutant M5 decreased up to 80%.

  9. Isolation of induced mutants using gamma ray and ethyl methane sulphonate in Tomato (Solanum lycopersicum L.

    Directory of Open Access Journals (Sweden)

    S. Sikder, V. K. Ravat, S. Basfore and P. Hazra

    2015-06-01

    Full Text Available Present investigation was undertaken to compare the mutagenic efficiency and effectiveness of gamma ray and ethyl methane sulphonate (EMS and to study the frequency and spectrum of macro-mutations in tomato. For this purpose, two cultivars of tomato having widely divergent place of origin viz., Patharkutchi of West Bengal, India and Alisa Craig of England were treated with 50, 100, 150, 200 and 250Gy gamma rays and 0.05, 0.10, 0.15, 0.20 and 0.25% (V/V EMS solution. Results showed that seed germination, seedling height and pollen fertility in M1 generation reduced steadily with the increasing doses of both mutagens. The LD50 dose for Patharkutchi and Alisa Craig was 310.7Gy and 229.7Gy gamma ray, 0.30% and 0.20% EMS concentration, respectively. Gamma ray (50Gy to 150Gy proved to be more efficient and effective mutagen followed by 0.05% to 10% EMS treatment. Five true breeding mutants hold promise for their utilization in tomato breeding programme.

  10. Gamma rays induced moisture stress tolerant long root mutant in mungbean (Vigna radiata L. Wilczek

    Directory of Open Access Journals (Sweden)

    V. J. Dhole And K. S. Reddy

    2010-09-01

    Full Text Available The mutation breeding programme was undertaken to improve the root characters of variety Samrat in mungbean. It is highyielding and MYMV resistant variety but its shallow root system limits the grain yield under moisture stress. Seeds of varietySamrat were irradiated with 450 Gy gamma rays. One long root mutant (LRM having root length up to 71cm as compared toaverage 19.5 cm of Samrat was identified and isolated in M2 generation. LRM was tested in laboratory using variousconcentrations of polyethylene glycol and also in the field under simulated water stress condition. With increase inconcentration of PEG, water uptake, germination, root and shoot length significantly reduced in both LRM and Samrat. TheLRM showed significantly greater water uptake, root and shoot length as compared to control Samrat. In the field experiment,LRM recorded better root length along with yield and yield contributing characters as compared to Samrat. The resultsindicated that a strong root system of LRM will help the genotype to draw comparatively more water to survive and reproducenormally under drought.

  11. Induced Levels of Heat Shock Proteins in dnaK mutants of Lactococcus lactis

    DEFF Research Database (Denmark)

    Koch, Birgit; Hammer, Karin; Vogensen, Finn K.

    1998-01-01

    of the inferred substrate binding site of the DnaK protein, exhibits a pronounced temperature sensitive phenotype and shows altered regulation of the heat shock response. The expression of the heat shock proteins are increased at the normal growth temperature measured both as protein synthesis rates and m......, HrcA , is dependent of the chaperone function of the GroELS complex and that an insertion mutant in dnaK did not have any effect on the expression of the heat shock proteins. The present data from Lactococcus lactis suggest that the DnaK protein could be involved in maturation of the homologous Hrc......The bacterial heat shock response is characterized by the elevated expression of a number of chaperone complexes and proteases including the DnaK-GrpE-DnaJ and the GroELS chaperone complexes. In order to investigate the importance of the DnaK chaperone complex for the growth and the heat shock...

  12. Targeted Inhibition of EGFR and Glutaminase Induces Metabolic Crisis in EGFR Mutant Lung Cancer.

    Science.gov (United States)

    Momcilovic, Milica; Bailey, Sean T; Lee, Jason T; Fishbein, Michael C; Magyar, Clara; Braas, Daniel; Graeber, Thomas; Jackson, Nicholas J; Czernin, Johannes; Emberley, Ethan; Gross, Matthew; Janes, Julie; Mackinnon, Andy; Pan, Alison; Rodriguez, Mirna; Works, Melissa; Zhang, Winter; Parlati, Francesco; Demo, Susan; Garon, Edward; Krysan, Kostyantyn; Walser, Tonya C; Dubinett, Steven M; Sadeghi, Saman; Christofk, Heather R; Shackelford, David B

    2017-01-17

    Cancer cells exhibit increased use of nutrients, including glucose and glutamine, to support the bioenergetic and biosynthetic demands of proliferation. We tested the small-molecule inhibitor of glutaminase CB-839 in combination with erlotinib on epidermal growth factor receptor (EGFR) mutant non-small cell lung cancer (NSCLC) as a therapeutic strategy to simultaneously impair cancer glucose and glutamine utilization and thereby suppress tumor growth. Here, we show that CB-839 cooperates with erlotinib to drive energetic stress and activate the AMP-activated protein kinase (AMPK) pathway in EGFR (del19) lung tumors. Tumor cells undergo metabolic crisis and cell death, resulting in rapid tumor regression in vivo in mouse NSCLC xenografts. Consistently, positron emission tomography (PET) imaging with (18)F-fluoro-2-deoxyglucose ((18)F-FDG) and (11)C-glutamine ((11)C-Gln) of xenografts indicated reduced glucose and glutamine uptake in tumors following treatment with CB-839 + erlotinib. Therefore, PET imaging with (18)F-FDG and (11)C-Gln tracers can be used to non-invasively measure metabolic response to CB-839 and erlotinib combination therapy. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

  13. Targeted Inhibition of EGFR and Glutaminase Induces Metabolic Crisis in EGFR Mutant Lung Cancer

    Directory of Open Access Journals (Sweden)

    Milica Momcilovic

    2017-01-01

    Full Text Available Cancer cells exhibit increased use of nutrients, including glucose and glutamine, to support the bioenergetic and biosynthetic demands of proliferation. We tested the small-molecule inhibitor of glutaminase CB-839 in combination with erlotinib on epidermal growth factor receptor (EGFR mutant non-small cell lung cancer (NSCLC as a therapeutic strategy to simultaneously impair cancer glucose and glutamine utilization and thereby suppress tumor growth. Here, we show that CB-839 cooperates with erlotinib to drive energetic stress and activate the AMP-activated protein kinase (AMPK pathway in EGFR (del19 lung tumors. Tumor cells undergo metabolic crisis and cell death, resulting in rapid tumor regression in vivo in mouse NSCLC xenografts. Consistently, positron emission tomography (PET imaging with 18F-fluoro-2-deoxyglucose (18F-FDG and 11C-glutamine (11C-Gln of xenografts indicated reduced glucose and glutamine uptake in tumors following treatment with CB-839 + erlotinib. Therefore, PET imaging with 18F-FDG and 11C-Gln tracers can be used to non-invasively measure metabolic response to CB-839 and erlotinib combination therapy.

  14. UNTAGGED MUTATION IN RICE GAL4/VP16 TRANSCRIPTIONAL ACTIVATOR FACILITATED-ENHANCER TRAP LINES

    Directory of Open Access Journals (Sweden)

    Sri Koerniati

    2013-04-01

    Full Text Available An enhancer trap system is an insertional mutagenesis based upon gene expression, instead of gene knock-out, so its insertion in genome is  expected not linked to any dramatic changes in plant phenotypes. Gene  knock-out, leading to lossof- function (LoF mutation, is a dominant  approach for rice functional genomic studies. The objective of this study was to find out whether Transcriptional Activator-Facilitated Enhancer Trap (TAFET T-DNA insertion inducing mutant phenotypes in rice TAFET population. Materials used in this experiment were T1 generation of 270 rice TAFET lines. Eight plants of each were grown in the greenhouse and observed for any mutant phenotypes. Phenotypic, histochemical, Southernblot analyses were carried out to define a mutant of pSKC66.1- 8e. Result showed that about 10% of the 270 lines produced chlorophyll-deficient  leaves, ranged from yellowish green (viridis, white stripe green zebra-like stripe to completely white (albino. Albino plants died after two weeks,  whilst white stripe or viridis mutants became normal in the next generation(T2. Another mutant was pSKC66.1-8e line which had floral dramatic phenotype change with various spikelet shapes and number of organs, and had a single twisted culm. The flower of mutant also had gus gene expression. Plants with wild type did not express gus gene and had six or more straight culms. Molecular, histochemical and phenotypic analyses of this particular line for three generations indicated that mutant phenotype was not due to the T-DNA insertion. Since there was approved that Tos17 is activated during tissue culture and induced mutant phenotype, this line might relate to Tos17 insertion, but it needs further investigation to gain such conclusion.

  15. Mutant KRas-Induced Mitochondrial Oxidative Stress in Acinar Cells Upregulates EGFR Signaling to Drive Formation of Pancreatic Precancerous Lesions

    Directory of Open Access Journals (Sweden)

    Geou-Yarh Liou

    2016-03-01

    Full Text Available The development of pancreatic cancer requires the acquisition of oncogenic KRas mutations and upregulation of growth factor signaling, but the relationship between these is not well established. Here, we show that mutant KRas alters mitochondrial metabolism in pancreatic acinar cells, resulting in increased generation of mitochondrial reactive oxygen species (mROS. Mitochondrial ROS then drives the dedifferentiation of acinar cells to a duct-like progenitor phenotype and progression to PanIN. This is mediated via the ROS-receptive kinase protein kinase D1 and the transcription factors NF-κB1 and NF-κB2, which upregulate expression of the epidermal growth factor, its ligands, and their sheddase ADAM17. In vivo, interception of KRas-mediated generation of mROS reduced the formation of pre-neoplastic lesions. Hence, our data provide insight into how oncogenic KRas interacts with growth factor signaling to induce the formation of pancreatic cancer.

  16. Characterization of an inositol 1,3,4-trisphosphate 5/6-kinase gene that is essential for drought and salt stress responses in rice.

    Science.gov (United States)

    Du, Hao; Liu, Linhong; You, Lei; Yang, Mei; He, Yubing; Li, Xianghua; Xiong, Lizhong

    2011-12-01

    Drought and salt stresses are major limiting factors for crop production. To identify critical genes for stress resistance in rice (Oryza sativa L.), we screened T-DNA mutants and identified a drought- and salt-hypersensitive mutant dsm3. The mutant phenotype was caused by a T-DNA insertion in a gene encoding a putative inositol 1,3,4-trisphosphate 5/6-kinase previously named OsITPK2 with unknown function. Under drought stress conditions, the mutant had significantly less accumulation of osmolytes such as proline and soluble sugar and showed significantly reduced root volume, spikelet fertility, biomass, and grain yield; however, malondialdehyde level was increased in the mutant. Interestingly, overexpression of DSM3 (OsITPK2) in rice resulted in drought- and salt-hypersensitive phenotypes and physiological changes similar to those in the mutant. Inositol trisphosphate (IP3) level was decreased in the overexpressors under normal condition and drought stress. A few genes related to osmotic adjustment and reactive oxygen species scavenging were down-regulated in the mutant and overexpression lines. The expression level of DSM3 promoter-driven β-glucuronidase (GUS) reporter gene in rice was induced by drought, salt and abscisic acid. Protoplast transient expression assay indicated that DSM3 is an endoplasmic reticulum protein. Sequence analysis revealed six putative ITPKs in rice. Transcript level analysis of OsITPK genes revealed that they had different tempo-spatial expression patterns, and the responses of DSM3 to abiotic stresses, including drought, salinity, cold, and high temperature, were distinct from the other five members in rice. These results together suggest that DSM3/OsITPK2 is an important member of the OsITPK family for stress responses, and an optimal expression level is essential for drought and salt tolerance in rice.

  17. An early response regulatory cluster induced by low temperature and hydrogen peroxide in seedlings of chilling-tolerant japonica rice

    Directory of Open Access Journals (Sweden)

    Jia Yulin

    2007-06-01

    Full Text Available Abstract Background Plants respond to low temperature through an intricately coordinated transcriptional network. The CBF/DREB-regulated network of genes has been shown to play a prominent role in freeze-tolerance of Arabidopsis through the process of cold acclimation (CA. Recent evidence also showed that the CBF/DREB regulon is not unique to CA but evolutionarily conserved between chilling-insensitive (temperate and chilling-sensitive (warm-season plants. In this study, the wide contrast in chilling sensitivity between indica and japonica rice was used as model to identify other regulatory clusters by integrative analysis of promoter architecture (ab initio and gene expression profiles. Results Transcriptome analysis in chilling tolerant japonica rice identified a subset of 121 'early response' genes that were upregulated during the initial 24 hours at 10°C. Among this group were four transcription factors including ROS-bZIP1 and another larger sub-group with a common feature of having as1/ocs-like elements in their promoters. Cold-induction of ROS-bZIP1 preceded the induction of as1/ocs-like element-containing genes and they were also induced by exogenous H2O2 at ambient temperature. Coordinated expression patterns and similar promoter architectures among the 'early response' genes suggest that they belong to a potential regulon (ROS-bZIP – as1/ocs regulatory module that responds to elevated levels of ROS during chilling stress. Cultivar-specific expression signatures of the candidate genes indicate a positive correlation between the activity of the putative regulon and genotypic variation in chilling tolerance. Conclusion A hypothetical model of an ROS-mediated regulon (ROS-bZIP – as1/ocs triggered by chilling stress was assembled in rice. Based on the current results, it appears that this regulon is independent of ABA and CBF/DREB, and that its activation has an important contribution in configuring the rapid responses of rice seedlings

  18. p53 mutant human glioma cells are sensitive to UV-C-induced apoptosis due to impaired cyclobutane pyrimidine dimer removal.

    Science.gov (United States)

    Batista, Luis F Z; Roos, Wynand P; Kaina, Bernd; Menck, Carlos F M

    2009-02-01

    The p53 protein is a key regulator of cell responses to DNA damage, and it has been shown that it sensitizes glioma cells to the alkylating agent temozolomide by up-regulating the extrinsic apoptotic pathway, whereas it increases the resistance to chloroethylating agents, such as ACNU and BCNU, probably by enhancing the efficiency of DNA repair. However, because these agents induce a wide variety of distinct DNA lesions, the direct importance of DNA repair is hard to access. Here, it is shown that the induction of photoproducts by UV light (UV-C) significantly induces apoptosis in a p53-mutated glioma background. This is caused by a reduced level of photoproduct repair, resulting in the persistence of DNA lesions in p53-mutated glioma cells. UV-C-induced apoptosis in p53 mutant glioma cells is preceded by strong transcription and replication inhibition due to blockage by unrepaired photolesions. Moreover, the results indicate that UV-C-induced apoptosis of p53 mutant glioma cells is executed through the intrinsic apoptotic pathway, with Bcl-2 degradation and sustained Bax and Bak up-regulation. Collectively, the data indicate that unrepaired DNA lesions induce apoptosis in p53 mutant gliomas despite the resistance of these gliomas to temozolomide, suggesting that efficiency of treatment of p53 mutant gliomas might be higher with agents that induce the formation of DNA lesions whose global genomic repair is dependent on p53.

  19. Heat-induced phytohormone changes are associated with disrupted early reproductive development and reduced yield in rice

    Science.gov (United States)

    Wu, Chao; Cui, Kehui; Wang, Wencheng; Li, Qian; Fahad, Shah; Hu, Qiuqian; Huang, Jianliang; Nie, Lixiao; Peng, Shaobing

    2016-01-01

    Heat stress causes morphological and physiological changes and reduces crop yield in rice (Oryza sativa). To investigate changes in phytohormones and their relationships with yield and other attributes under heat stress, four rice varieties (Nagina22, Huanghuazhan, Liangyoupeijiu, and Shanyou 63) were grown in pots and subjected to three high temperature treatments plus control in temperature-controlled greenhouses for 15 d during the early reproductive phase. Yield reductions in Nagina22, Huanghuazhan, and Liangyoupeijiu were attributed to reductions in spikelet fertility, spikelets per panicle, and grain weight. The adverse effects of high temperature were alleviated by application of exogenous 6-benzylaminopurine (6-BA) in the heat-susceptible Liangyoupeijiu. High temperature stress reduced active cytokinins, gibberellin A1 (GA1), and indole-3-acetic acid (IAA), but increased abscisic acid (ABA) and bound cytokinins in young panicles. Correlation analyses and application of exogenous 6-BA revealed that high temperature-induced cytokinin changes may regulate yield components by modulating the differentiation and degradation of branches and spikelets, panicle exsertion, pollen vigor, anther dehiscence, and grain size. Heat-tolerant Shanyou 63 displayed minor changes in phytohormones, panicle formation, and grain yield under high temperature compared with those of the other three varieties. These results suggest that phytohormone changes are closely associated with yield formation, and a small reduction or stability in phytohormone content is required to avoid large yield losses under heat stress. PMID:27713528

  20. Anticarcinogenic efficacy of phytic acid extracted from rice bran on azoxymethane-induced colon carcinogenesis in rats.

    Science.gov (United States)

    Norazalina, S; Norhaizan, M E; Hairuszah, I; Norashareena, M S

    2010-05-01

    This study is carried out to determine the potential of phytic acid extracted from rice bran in the suppression of colon carcinogenesis induced by azoxymethane (AOM) in rats. Seventy-two male Sprague-Dawley rats were divided into 6 groups with 12 rats in each group. The intended rats for cancer treatment received two intraperitoneal injections of AOM in saline (15mg/kg bodyweight) over a 2-week period. The treatments of phytic acid were given in two concentrations: 0.2% (w/v) and 0.5% (w/v) during the post-initiation phase of carcinogenesis phase via drinking water. The colons of the animals were analyzed for detection and quantification of aberrant crypt foci (ACF) after 8 weeks of treatment. The finding showed treatment with 0.2% (w/v) extract phytic acid (EPA) gave the greatest reduction in the formation of ACF. In addition, phytic acid significantly suppressed the number of ACF in the distal, middle and proximal colon as compared to AOM alone (pphytic acid (CPA) had the highest percentage (71%) of non-dysplastic ACF followed by treatment with 0.2% (w/v) EPA (61%). Administration of phytic acid also reduced the incidence and multiplicity of total tumors even though there were no significant differences between groups. In conclusion, this study found the potential value of phytic acid extracted from rice bran in reducing colon cancer risk in rats.

  1. Replication-competent, oncolytic herpes simplex virus type 1 mutants induce a bystander effect following ganciclovir treatment.

    Science.gov (United States)

    Luo, Chenhong; Mori, Isamu; Goshima, Fumi; Ushijima, Yoko; Nawa, Akihiro; Kimura, Hiroshi; Nishiyama, Yukihiro

    2007-10-01

    Cells expressing herpes simplex virus (HSV) thymidine kinase (tk) are killed by ganciclovir (GCV). Adjacent cells without HSV-tk also die, a phenomenon known as the 'bystander effect'. However, there is no evidence that replication-competent HSV induces a bystander effect in the presence of GCV. Therefore, we investigated the bystander effect in HEp-2 cells infected with replication-competent, oncolytic HSV-1 mutants, hrR3 and HF10. In cells infected at a multiplicity of infection (MOI) of 3, GCV did not induce apoptosis. At low MOIs of 0.3 and 0.03, however, a number of adjacent, uninfected cells apoptosed following GCV treatment. Irrespective of GCV treatment, HEp-2 cells expressed minimal levels of connexin 43 (Cx43). However, Cx43 expression was enhanced by GCV in response to infection with HF10 at an MOI of 0.3, but not at an MOI of 3. Expression of other proteins involved in gap junctions, including Cx26 and Cx40, was not augmented under these conditions. The PKA and PI3K signal transduction pathways are likely involved in enhanced Cx43 expression as inhibitors of these pathways prevented Cx43 upregulation. These results suggest that infection with replication-competent HSV-1 induces the bystander effect in cells treated with GCV because of efficient intercellular transport of active GCV through abundant gap junctions. Copyright 2007 John Wiley & Sons, Ltd.

  2. Genetic and molecular dissection of naturally occurring variations in rice

    Energy Technology Data Exchange (ETDEWEB)

    Yano, Masahiro [National Institute of Agrobiological Sciences, Tsukuba, Ibaraki (Japan)

    2002-02-01

    The progress for structural analysis of the rice genome has allowed us to embark on the sequencing of the whole rice genome. Resources - genetic markers, sequence data, and genomic clones - derived from many efforts will be used for the functional analysis of rice genes in the next decade. Although artificially induced variations, such as mutants, have been used mainly for genetic and physiological studies in rice and other plant species, the development of DNA markers has made possible access to naturally occurring allelic variations underlying complex traits. Such analysis is often referred to as quantitative traits locus (QTL) analysis. Many QTLs have been mapped for many complex traits in rice. During the analyses of several quantitative traits by the DNA marker-assisted strategy, two questions about QTL analysis have been raised: 1) Does a QTL represent a single Mendelian locus or a cluster of multiple loci? 2) Is it possible to precisely map a QTL and identify QTLs at the molecular level using map-based or other strategies? To answer these questions, a series of analyses on heading date, including the identification of putative QTLs, characterization and fine mapping of QTLs using nearly isogenic lines (NILs), and identification of genes at QTLs for heading date by the map-based strategy has been performed. In addition, several primary permanent mapping populations and secondary genetic resources, such as chromosomal segmental substitution lines, have been developed to facilitate the genetic analysis of naturally occurring allelic variation. (M. Suetake)

  3. Nimotuzumab enhances temozolomide-induced growth suppression of glioma cells expressing mutant EGFR in vivo.

    Science.gov (United States)

    Nitta, Yusuke; Shimizu, Saki; Shishido-Hara, Yukiko; Suzuki, Kaori; Shiokawa, Yoshiaki; Nagane, Motoo

    2016-03-01

    A mutant form of epidermal growth factor receptor (EGFR), EGFRvIII, is common in glioblastoma (GBM) and confers enhanced tumorigenic activity and drug resistance. Nimotuzumab, an anti-EGFR antibody, has shown preclinical and clinical activity to GBM, but its specific activity against EGFRvIII has not been fully investigated. Human glioma U87MG or LNZ308 cells overexpressing either wild-type (wt) EGFR or EGFRvIII were treated with nimotuzumab, temozolomide, or both. Expression and phosphorylation status of molecules were determined by Western blot analysis. Methylation status of promoter region of O(6) -methylguanine-DNA methyltransferase (MGMT) was detected by methylation-specific PCR. Antitumor activity was tested using nude mice bearing either subcutaneous or intracerebral xenografts along with analyses of EGFR phosphorylation status, proliferation, apoptosis, and vessel density. Nimotuzumab treatment resulted in reduction of EGFRvIII tyrosine phosphorylation with a decrease in Akt phosphorylation that was greater than that of wtEGFR. Correspondingly, antitumor effects, growth suppression and survival elongation, were more significant in mice bearing either subcutaneous or intracerebral tumor expressing EGFRvIII than in those expressing wtEGFR. These effects were markedly increased when temozolomide was combined with nimotuzumab. The post-treatment recurrent brain tumors exhibited a decrease in expression of the mismatch repair (MMR) proteins, MSH6 and MLH1, but their methylated MGMT status did not changed. Nimotuzumab has in vivo antitumor activity against GBM, especially those expressing EGFRvIII, when combined with temozolomide. This could provide a basis for preselection of patients with GBM by EGFR status who might benefit from the nimotuzumab and temozolomide combination therapy.

  4. Ontogenetically-regulated male sterility in tissue culture - induced and spontaneous sorghum mutants

    Directory of Open Access Journals (Sweden)

    Elkonin L.A.

    2003-01-01

    Full Text Available Variability of male fertility expression in the AS-1 line, a somaclonal variant obtained from tissue culture of CMS-plant, and in the progeny of revenant '124-1' obtained from fertile tiller, which developed on CMS-plant transferred from the field to the greenhouse, was investigated. Both revertants were characterized by similar expression of male fertility during plant ontogenesis: the panicle on the main tiller was almost completely sterile whereas formation of fertile pollen grains and seed set were observed on the panicles of the shoot tillers. A clear basipetal gradient of male fertility was manifested on all panicles: the base had significantly higher per cent of fertile pollen grains in comparison with the middle part, while in the top the anthers were either absent or had few sterile pollen grains. Such an ontogenetically-regulated restoration of male fertility was controlled by nuclear genes and could be transferred through the pollen in crosses with progenitor CMS-line. Growing of AS-1 plants in the growth chambers simultaneously under a long (16/8 and a short (12/12 daylength conditions demonstrated that differences of fertility level in different tillers was not caused by change of photoperiod during plant ontogenesis and functioning of photoperiod-sensitive fertility restoring gene. Whereas, the ontogenetically-regulated expression of male fertility in both revenants was temperature-dependent and was clearly manifested under relatively cool conditions during 2-week period before the beginning of anthesis of the first panicle (average daily temperature 21°C. The increase of the average daily temperature by 2-3 С resulted in sharp increase of male fertility level. Possibility of using AS-1 line in a new "two-line system" of hybrid seed production, which require only two lines (sterile mutant and fertility restorer, is discussed.

  5. Impact of Non-Invasively Induced Motor Deficits on Tibial Cortical Properties in Mutant Lurcher Mice.

    Directory of Open Access Journals (Sweden)

    Alena Jindrová

    Full Text Available It has been shown that Lurcher mutant mice have significantly altered motor abilities, regarding their motor coordination and muscular strength because of olivorecebellar degeneration. We assessed the response of the cross-sectional geometry and lacuno-canalicular network properties of the tibial mid-diaphyseal cortical bone to motor differences between Lurcher and wild-type (WT male mice from the B6CBA strain. The first data set used in the cross-sectional geometry analysis consists of 16 mice of 4 months of age and 32 mice of 9 months of age. The second data set used in the lacunar-canalicular network analysis consists of 10 mice of 4 months of age. We compared two cross-sectional geometry and four lacunar-canalicular properties by I-region using the maximum and minimum second moment of area and anatomical orientation as well as H-regions using histological differences within a cross section. We identified inconsistent differences in the studied cross-sectional geometry properties between Lurcher and WT mice. The biggest significant difference between Lurcher and WT mice is found in the number of canaliculi, whereas in the other studied properties are only limited. Lurcher mice exhibit an increased number of canaliculi (p < 0.01 in all studied regions compared with the WT controls. The number of canaliculi is also negatively correlated with the distance from the centroid in the Lurcher and positively correlated in the WT mice. When the Lurcher and WT sample is pooled, the number of canaliculi and lacunar volume is increased in the posterior Imax region, and in addition, midcortical H-region exhibit lower number of canaliculi, lacuna to lacuna distance and increased lacunar volume. Our results indicate, that the importance of precise sample selection within cross sections in future studies is highlighted because of the histological heterogeneity of lacunar-canalicular network properties within the I-region and H-region in the mouse cortical

  6. Note on early testing of induced mutants of chickpea under rainfed and irrigated conditions

    Energy Technology Data Exchange (ETDEWEB)

    Nerkar, Y.S.; Patil, V.D. (Marathwada Agricultural Univ., Parbhani (India))

    1981-09-01

    Gamma-ray-induced mutations involving yield and yield-contributing characters were isolated in the M/sub 3/ and M/sub 4/ generations of 'Chaffa', 'N 59' and 'No. 82-3-50' varieties of chickpea (Cicer arientinum Linn.).

  7. Characterization of Arabidopsis enhanced disease susceptibility mutants that are affected in systemically induced resistance

    NARCIS (Netherlands)

    Ton, J.; Vos, M. de; Robben, C.; Buchala, Anthony; Métraux, Jean-Pierre; Loon, L.C. van; Pieterse, C.M.J.

    2002-01-01

    In Arabidopsis, the rhizobacterial strain Pseudomonas fluorescens WCS417r triggers jasmonate (JA)- and ethylene (ET)-dependent induced systemic resistance (ISR) that is effective against different pathogens. Arabidopsis genotypes unable to express rhizobacteria-mediated ISR against the bacterial pat

  8. A mutant prion protein sensitizes neurons to glutamate-induced excitotoxicity.

    Science.gov (United States)

    Biasini, Emiliano; Unterberger, Ursula; Solomon, Isaac H; Massignan, Tania; Senatore, Assunta; Bian, Hejiao; Voigtlaender, Till; Bowman, Frederick P; Bonetto, Valentina; Chiesa, Roberto; Luebke, Jennifer; Toselli, Paul; Harris, David A

    2013-02-06

    Growing evidence suggests that a physiological activity of the cellular prion protein (PrP(C)) plays a crucial role in several neurodegenerative disorders, including prion and Alzheimer's diseases. However, how the functional activity of PrP(C) is subverted to deliver neurotoxic signals remains uncertain. Transgenic (Tg) mice expressing PrP with a deletion of residues 105-125 in the central region (referred to as ΔCR PrP) provide important insights into this problem. Tg(ΔCR) mice exhibit neonatal lethality and massive degeneration of cerebellar granule neurons, a phenotype that is dose dependently suppressed by the presence of wild-type PrP. When expressed in cultured cells, ΔCR PrP induces large, ionic currents that can be detected by patch-clamping techniques. Here, we tested the hypothesis that abnormal ion channel activity underlies the neuronal death seen in Tg(ΔCR) mice. We find that ΔCR PrP induces abnormal ionic currents in neurons in culture and in cerebellar slices and that this activity sensitizes the neurons to glutamate-induced, calcium-mediated death. In combination with ultrastructural and biochemical analyses, these results demonstrate a role for glutamate-induced excitotoxicity in PrP-mediated neurodegeneration. A similar mechanism may operate in other neurodegenerative disorders attributable to toxic, β-rich oligomers that bind to PrP(C).

  9. Induced Pib Expression and Resistance to Magnaporthe grisea are Compromised by Cytosine Demethylation at Critical Promoter Regions in Rice

    Institute of Scientific and Technical Information of China (English)

    Yuan Li; Qiong Xia; Hongping Kou; Dan Wang; Xiuyun Lin; Ying Wu; Chunming Xu; Shaochen Xing

    2011-01-01

    Pib is a well-characterized rice blast-resistance gene belonging to the nucleotide binding site (NBS) and leucine-rich repeat (LRR) superfamily.Expression of Pib was low under non-challenged conditions,but strongly induced by the blast-causing fungal pathogen Magnaporthe grisea,thereby conferring resistance to the pathogen.It is generally established that cytosine methylation of the promoter-region often plays a repressive role in modulating expression of the gene in question.We report here that two critical regions of the Pib promoter were heavily CG cytosine-methylated in both cultivars studied.Surprisingly,induced expression of Pib by M.grisea infection did not entail its promoter demethylation,and partial demethylation by 5-azacytidine-treatment actually reduced Pib expression relative to wildtype plants.Accordingly,the blast disease-resistance was compromised in the 5’-azaC-treated plants relative to wild-type.In contrast,the disease susceptibility was not affected by the 5’-azaC treatment in another two rice cultivars that did not contain the Pib gene,ruling out effects of other R genes and non-specific genotoxic effects by the drug-treatment as a cause for the compromised Pib-conditioned blast-resistance.Taken together,our results suggest that promoter DNA methylation plays a novel enhancing role in conditioning high-level of induced expression of the Pib gene in times of M.grisea infection,and its conferred resistance to the pathogen.

  10. Effect of pre-germinated brown rice intake on diabetic neuropathy in streptozotocin-induced diabetic rats

    Directory of Open Access Journals (Sweden)

    Ariga Toshio

    2007-11-01

    Full Text Available Abstract Background To study the effects of a pre-germinated brown rice diet (PR on diabetic neuropathy in streptozotocin (STZ-induced diabetic rats. Methods The effects of a PR diet on diabetic neuropathy in STZ-induced diabetic rats were evaluated and compared with those fed brown rice (BR or white rice (WR diets with respect to the following parameters: blood-glucose level, motor-nerve conduction velocity (NCV, sciatic-nerve Na+/K+-ATPase activity, and serum homocysteine-thiolactonase (HTase activity. Results Compared with diabetic rats fed BR or WR diets, those fed a PR diet demonstrated significantly lower blood-glucose levels (p +/K+-ATPase activity (1.6- and 1.7-fold higher, respectively. The PR diet was also able to normalize decreased serum homocysteine levels normally seen in diabetic rats. The increased Na+/K+-ATPase activity observed in rats fed PR diets was associated with elevations in HTase activity (r = 0.913, p in vitro effect of the total lipid extract from PR bran (TLp on the Na+/K+-ATPase and HTase activity was also examined. Incubation of homocysteine thiolactone (HT with low-density lipoprotein (LDL in vitro resulted in generation of HT-modified LDL, which possessed high potency to inhibit Na+/K+-ATPase activity in the sciatic nerve membrane. The inhibitory effect of HT-modified LDL on Na+/K+-ATPase activity disappeared when TLp was added to the incubation mixture. Furthermore, TLp directly activated the HTase associated with high-density lipoprotein (HDL. Conclusion PR treatment shows efficacy for protecting diabetic deterioration and for improving physiological parameters of diabetic neuropathy in rats, as compared with a BR or WR diet. This effect may be induced by a mechanism whereby PR intake mitigates diabetic neuropathy by one or more factors in the total lipid fraction. The active lipid fraction is able to protect the Na+/K+-ATPase of the sciatic-nerve membrane from the toxicity of HT-modified LDL and to directly

  11. Review on Mutation in Lateral Root of Rice

    Institute of Scientific and Technical Information of China (English)

    ZHANG Xia; ZHANG Da; HAO Zaibin

    2011-01-01

    Rice roots include seminal roots, adventitious roots, lateral roots and root hairs, At present, progresses in the research of rice roots have been achieved in many aspects, such as root morphology, root activity, root reaction to various environmental factors as a contribution of root growth and rice yield, the relationship between root growth and stems/leaves/flowers/rice, genetic laws of root characters, etc. However, there are very few researches on lateral root mutant. This paper reviewed progresses of the lateral root mutant of rice from the perspectives of phytomorphology to plant physiology and biochemistry to the gene mapping, consisting of mechanism of developing lateral root of rice, gene cloning and functional analysis of lateral root development, the relationship between auxin and lateral roots, agronomic traits of lateral roots mutant, structure and morphology of root hairs, gravity anomaly of root, redox metabolism and proteomics researches of the mutation in lateral root of rice.

  12. 水稻类树状突变体s2-21的遗传分析与精细定位%Genetic Analysis and Gene Fine Mapping of Rice Leafy Head Mutant s2-21

    Institute of Scientific and Technical Information of China (English)

    王海凤; 高洁; 孙伟; 张士永; 赵庆雷; 尹亮; 赵金凤; 李学勇; 袁守江

    2014-01-01

    叶序和出叶间隔期是叶片生长发育的基本生物学特性和水稻的重要农艺性状之一。对叶序或出叶间隔期突变体的研究,可以帮助我们了解叶片的形成机制。本研究通过甲基磺酸乙酯(EMS)诱变粳稻品种日本晴,获得一个稳定遗传的类树状突变体s2-21。该突变体出叶间隔期变短、节间缩短、植株矮化、分蘖数减少、叶片数增加、不能正常进行生殖生长。将该突变体与籼稻品种Dular杂交,遗传分析表明该突变体性状受1对隐性基因控制。通过InDel分子标记对s2-21/Dular F2群体进行遗传定位,将该基因初步定位在第1染色体InDel标记C1-15和S1-17之间。利用本实验已测序的籼稻品种Dular全基因组序列与NCBI (http://www.ncbi.nlm.nih.gov/)上提供的粳稻品种日本晴基因组序列比对,开发了6个新的InDel标记,最终将该基因定位在W25和W26之间约88 kb的区间内。测序结果表明该突变体中PLA2基因的第4个内含子的第5位碱基由G突变为A。%Phyllotaxy and plastochron are basic aspects of leaf development and the important agronomic traits of rice. The study of phyllotaxy or plastochron mutant will help us to understand the mechanism of leaf formation. In this study, a leafy head mutant s2-21 was isolated from an EMS (ethyl methane sulfonate) mutagenized japonica cultivar Nipponbare. It was characterized by shorter intervals between leaves, shortened internodes, dwarf plants, decreased tiller number, and increased leaf number. It was also unable to transit from vegetative to reproductive growth. Genetic analysis with a F2 population of the mutant and an indica cultivar Dular showed that this trait is controlled by a single recessive nucleus gene, which was primarily mapped between two insertion-deletion (InDel) markers C1-15 and S1-17 on chromosome 1. For the fine mapping, new InDel markers were designed by utilizing information of genomic sequences from

  13. 水稻穗颈极度缩短突变体sui(t)的遗传分析与基因定位%Genetic Analysis and Gene Mapping of Extremely Shortened Uppermost Internode Mutant sui(t) in Rice

    Institute of Scientific and Technical Information of China (English)

    王晓雯; 苗润隆; 蒋钰东; 杜川; 杨正林; 凌英华; 何光华; 桑贤春

    2013-01-01

    One novel mutant,tentatively designated as sui(t),was discovered from the progeny of indica rice restorer line Jinhui10,whose seeds were treated with EMS (ethyl methane sulfonate).This mutant was characterized by fully sheathed panicle and was insensitive to GA3.Morphology analysis showed that its uppermost internode was greatly shortened while the other internodes were normal.Genetic analysis suggested that the mutational character was controlled by a single recessive gene.Zhonghua11 was crossed with sui(t) and the F2 population was then used for gene mapping.SUI(t) gene was mapped between the SSR markers RM5336/RM3425 and the tag end of the short arm of Chromosome Ⅰ.SSR and Indel markers were designed and S UI(t) gene was finally restricted within the region of RM3148 and ClIn002 (swu),with a genetic distance of 0.8 cM and 1.4 cM,respectively,and a physical distance of 382 kb.The above result provides a foundation for SUI(t) gene cloning and functional analysis.%来源于EMS处理籼型恢复系缙恢10号的穗颈节间极度缩短突变体其他节间与恢复系缙恢10号无显著变化,表现为全包穗,对GA3不敏感,暂命名为sui(t).遗传分析表明变异性状受一对隐性核基因控制.以中花11/sui(t)的F2隐性单株为定位群体,将sui(t)基因定位在第1染色体SSR标记RM5336,RM3425向短臂末端一侧,进一步合成SSR,Indel等标记,最终将sui(t)基因定位在RM3148和C1 In002(swu)之间,遗传距离分别为0.8 cM和1.4 cM,物理距离为382 kb,为下一步基因克隆和功能研究奠定了基础.

  14. Mutagenic effects of heavy ion irradiation on rice seeds

    Energy Technology Data Exchange (ETDEWEB)

    Xu Xue [School of Agronomy, Anhui Agricultural University, 130 Changjiang West Road, Hefei 230036 (China); Key Laboratory of Ion Beam Bio-Engineering, Institute of Technical Biology and Agriculture Engineering, 350 Shushanhu Road, Hefei 230031 (China); Liu Binmei; Zhang Lili [Key Laboratory of Ion Beam Bio-Engineering, Institute of Technical Biology and Agriculture Engineering, 350 Shushanhu Road, Hefei 230031 (China); Wu Yuejin, E-mail: yjwu@ipp.ac.cn [Key Laboratory of Ion Beam Bio-Engineering, Institute of Technical Biology and Agriculture Engineering, 350 Shushanhu Road, Hefei 230031 (China)

    2012-11-01

    Three varieties of rice seeds were subjected to irradiation using low-energy and medium-energy ions. The damage and mutations induced by the ions were examined. In addition, genetic analysis and gene mapping of spotted leaf (spl) mutants were performed. Low-energy ions had no significant influence on germination, survival or seedling height, except for the survival of Nipponbare. Medium-energy ions had a significant influence on germination and survival but had no significant effect on seedling height. In the low-energy group, among 60,000 M{sub 2} plants, 2823 putative morphological mutants were found, and the mutation frequency was approximately 4.71%. In the medium-energy group, 3132 putative morphological mutants were found, and the mutation frequency was approximately 5.22%. Five spl mutants (spl29-spl33) were obtained by ion irradiation, and the heredity of the spl mutants was stable. The characteristics of the spl mutants were found, by genetic analysis and preliminary mapping, to be controlled by a single recessive gene, and spl30 and spl33 were found to be new lesion-mimic mutants.

  15. Effects of mannitol induced osmotic stress on proline accumulation, pigment degradation, photosynthetic abilities and growth characters in C3 rice and C4 sorghum

    Institute of Scientific and Technical Information of China (English)

    Suriyan CHA-UM; Souvanh THADAVONG; Chalermpol KIRDMANEE

    2009-01-01

    Osmotic stress is one of the most important abiotic factors which inhibit growth and development in both the vegetative and reproductive stages of many plant species. The aim of this investigation was to compare the biochemical and physiological responses in C3 rice and C4 sorghum to water deficit. Chlorophyll a (Chla), chlorophyll b (Chlb), total chlorophyll (TC) and total carotenoid (Cx+c) contents in both rice and sorghum seedlings under osmotic stress were adversely affected, related to increasing osmotic pressure in the culture media. In addition, the chlorophyll's fluorescence parameters and net photosynthetic rate (Pn) decreased, leading to growth reduction. Also, a positive correlation was found between physiological and biochemical data, while proline accumulation showed a negative relationship. The Chlb, Pn and fresh weight were maintained better in osmotic-stressed (-1.205 MPa) C4 sorghum seedlings than those in C3 rice seedlings. The growth and physiological responses of C3 rice and C4 sorghum decreased depending on the plant species, the osmotic pressure in the media and their interactions. Pigment content and Pn ability in C4 sorghum grown under mannitol-induced osmotic stress increased to a greater degree than in C3 rice, resulting in maintenance of growth.

  16. Biochemical characterization of N-methyl N' -nitro-N-nitrosoguanidine-induced cadmium resistant mutants of Aspergillus niger

    Indian Academy of Sciences (India)

    Samar Kumar Pal; Tapan Kumar Das

    2005-12-01

    Two cadmium resistant mutants (Cd1 and Cd2) of Aspergillus niger, among the six isolated by mutagenization with N-methyl N′-nitro-N-nitrosoguanidine (MNNG) at pH 6.4 were selected for the study. Analysis of lipid composition of the mutants and the wildtype indicated that total lipid as well as individual lipids of the cadmium resistant mutants were changed as compared with that of the wildtype. The increased activities of metal-lothionein and reduced activities of D-xylose isomerase and L-phenylalanine ammonia lyase in cell free extract of the cadmium resistant mutants suggested that mutants could allow high concentration of cadmium salt as compared with that of the wildtype. The respiratory activity and intracellular as well as extracellular Cd2+ concentration of the mutants reflected the high tolerance of the Cd mutants to cadmium ion.

  17. Endogenous nitric oxide mediates alleviation of cadmium toxicity induced by calcium in rice seedlings

    Institute of Scientific and Technical Information of China (English)

    Long Zhang; Zhen Chen; Cheng Zhu

    2012-01-01

    The effect of calcium chloride (CaCl2) on rice seedling growth under cadmium chloride (CdCl2) stress,as well as the possible role of endogenous nitric oxide (NO) in this process,was studied.The growth of rice seedlings was seriously inhibited by CdCl2,and the inhibition was significantly mitigated by CaCl2.However,hemoglobin (Hb) and 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline1-oxyl-3-oxide (cPTIO) weakened the promotion effect of CaCl2.The resuhs of NO fluorescence localization suggest that growth accelerated by CaCl2 might be associated with elevated NO levels.The content of Cd,protein thiols (PBT),and nonprotein thiols (NPT) in cell walls,cell organelles,and soluble fractions,respectively,of rice seedlings decreased considerably in the presence of CaCl2,whereas the content of pectin,hemicellulose 1 (HC1),and hemicellulose 2 (HC2) increased significantly.Elimination of endogenous NO in Cd+Ca treatment could promote the transportation of Cd2+ to cell organelles and soluble fractions and increase the content of NPT and PBT in leaves.In addition,transportation of Cd2+ to cell organelles and soluble fractions was retarded in roots,the content of NPT increased,and the content of PBT decreased.With elimination of endogenous NO in Cd+Ca treatment,the content of pectin,HC 1,and HC2 decreased significantly.Thus,Ca may alleviate Cd toxicity via endogenous NO with variation in the levels of NPT,PBT,and matrix polysaccharides.

  18. Prevalence of Food Impaction-Induced Periodontitis in Conventionally Housed Marsh Rice Rats (Oryzomys palustris).

    Science.gov (United States)

    Messer, Jonathan G; Jiron, Jessica M; Chen, Hung-Yuan; Castillo, Evelyn J; Mendieta Calle, Jorge L; Reinhard, Mary K; Kimmel, Donald B; Aguirre, J Ignacio

    2017-02-01

    Marsh rice rats (Oryzomys palustris) fed a pelleted diet high in sucrose and casein have been used as a model for moderate to severe periodontitis. Here we characterize the prevalence, location, and histopathologic features of food-impaction lesions (FIL), a unique type of oral event, in rice rats fed standard pelleted rodent chow from weaning until 34 wk of age. Healthy female rats (n = 90; age, 4 wk) were weaned into groups (n = 10 to 24) and were euthanized at 4, 16, 22, 28, or 34 wk of age. At necropsy, high-resolution photographs of the 4 jaw quadrants were examined by 3 independent observers to determine the presence, number, and location of FIL. In addition, gross periodontitis was scored (scale, 0 to 4), and the hemimaxillar surface area containing FIL was measured. Serial sections of decalcified jaws were assessed histologically. The prevalence of FIL increased with age, and was 0% (baseline), 59.1%, 69.6%, 81.8% and 80.0% in rats at age 4, 16, 22, 28, and 34 wk, respectively. FIL were predominantly located (93.9%) in the maxillary palatal surfaces of the interproximal area between molars 2 and 3 and did not affect mandibular surfaces. The percentage of the hemimaxillar surface area occupied by FIL was 6.83%, 4.82%, 2.88%, and 6.52% in rats at age 16, 22, 28, and 34 wk, respectively. Histopathologic changes in FIL varied from localized gingivitis to larger, localized periodontitis-like lesions. These data indicate that FIL are common in rice rats fed standard rodent chow, are slight to mild in severity, and are localized to specific regions in the oral cavity, thus suggesting they may be a suitable model for local maxillary periodontitis when fed standard rodent chow.

  19. Systems-level analysis of nitrogen starvation-induced modifications of carbon metabolism in a Chlamydomonas reinhardtii starchless mutant.

    Science.gov (United States)

    Blaby, Ian K; Glaesener, Anne G; Mettler, Tabea; Fitz-Gibbon, Sorel T; Gallaher, Sean D; Liu, Bensheng; Boyle, Nanette R; Kropat, Janette; Stitt, Mark; Johnson, Shannon; Benning, Christoph; Pellegrini, Matteo; Casero, David; Merchant, Sabeeha S

    2013-11-01

    To understand the molecular basis underlying increased triacylglycerol (TAG) accumulation in starchless (sta) Chlamydomonas reinhardtii mutants, we undertook comparative time-course transcriptomics of strains CC-4348 (sta6 mutant), CC-4349, a cell wall-deficient (cw) strain purported to represent the parental STA6 strain, and three independent STA6 strains generated by complementation of sta6 (CC-4565/STA6-C2, CC-4566/STA6-C4, and CC-4567/STA6-C6) in the context of N deprivation. Despite N starvation-induced dramatic remodeling of the transcriptome, there were relatively few differences (5 × 10(2)) observed between sta6 and STA6, the most dramatic of which were increased abundance of transcripts encoding key regulated or rate-limiting steps in central carbon metabolism, specifically isocitrate lyase, malate synthase, transaldolase, fructose bisphosphatase and phosphoenolpyruvate carboxykinase (encoded by ICL1, MAS1, TAL1, FBP1, and PCK1 respectively), suggestive of increased carbon movement toward hexose-phosphate in sta6 by upregulation of the glyoxylate pathway and gluconeogenesis. Enzyme assays validated the increase in isocitrate lyase and malate synthase activities. Targeted metabolite analysis indicated increased succinate, malate, and Glc-6-P and decreased Fru-1,6-bisphosphate, illustrating the effect of these changes. Comparisons of independent data sets in multiple strains allowed the delineation of a sequence of events in the global N starvation response in C. reinhardtii, starting within minutes with the upregulation of alternative N assimilation routes and carbohydrate synthesis and subsequently a more gradual upregulation of genes encoding enzymes of TAG synthesis. Finally, genome resequencing analysis indicated that (1) the deletion in sta6 extends into the neighboring gene encoding respiratory burst oxidase, and (2) a commonly used STA6 strain (CC-4349) as well as the sequenced reference (CC-503) are not congenic with respect to sta6 (CC-4348

  20. Vaccination with non-toxic mutant toxic shock syndrome toxin-1 induces IL-17-dependent protection against Staphylococcus aureus infection.

    Science.gov (United States)

    Narita, Kouji; Hu, Dong-Liang; Asano, Krisana; Nakane, Akio

    2015-06-01

    Toxic shock syndrome toxin-1 (TSST-1) is one of superantigens produced by Staphylococcus aureus. We have previously demonstrated that vaccination with non-toxic mutant TSST-1 (mTSST-1) develops host protection to lethal S. aureus infection in mice. However, the detailed mechanism underlying this protection is necessary to elucidate because the passive transfer of antibodies against TSST-1 fails to provide complete protection against S. aureus infection. In this study, the results showed that interleukin-17A (IL-17A)-producing cells were increased in the spleen cells of mTSST-1-vaccinated mice. The main source of IL-17A in mTSST-1-vaccinated mice was T-helper 17 (Th17) cells. The protective effect of vaccination was induced when the vaccinated wild type but not IL-17A-deficient mice were challenged with S. aureus. Gene expression of chemokines, CCL2 and CXCL1, and infiltration of neutrophils and macrophages were increased in spleens and livers of vaccinated mice after infection. The IL-17A-dependent immune response was TSST-1 specific because TSST-1-deficient S. aureus failed to induce the response. The present study suggests that mTSST-1 vaccination is able to provide the IL-17A-dependent host defense against S. aureus infection which promotes chemokine-mediated infiltration of phagocytes into the infectious foci.

  1. BRAF inhibitors induce metastasis in RAS mutant or inhibitor-resistant melanoma cells by reactivating MEK and ERK signaling.

    Science.gov (United States)

    Sanchez-Laorden, Berta; Viros, Amaya; Girotti, Maria Romina; Pedersen, Malin; Saturno, Grazia; Zambon, Alfonso; Niculescu-Duvaz, Dan; Turajlic, Samra; Hayes, Andrew; Gore, Martin; Larkin, James; Lorigan, Paul; Cook, Martin; Springer, Caroline; Marais, Richard

    2014-03-25

    Melanoma is a highly metastatic and lethal form of skin cancer. The protein kinase BRAF is mutated in about 40% of melanomas, and BRAF inhibitors improve progression-free and overall survival in these patients. However, after a relatively short period of disease control, most patients develop resistance because of reactivation of the RAF-ERK (extracellular signal-regulated kinase) pathway, mediated in many cases by mutations in RAS. We found that BRAF inhibition induces invasion and metastasis in RAS mutant melanoma cells through a mechanism mediated by the reactivation of the MEK (mitogen-activated protein kinase kinase)-ERK pathway, increased expression and secretion of interleukin 8, and induction of protease-dependent invasion. These events were accompanied by a cell morphology switch from predominantly rounded to predominantly elongated cells. We also observed similar responses in BRAF inhibitor-resistant melanoma cells. These data show that BRAF inhibitors can induce melanoma cell invasion and metastasis in tumors that develop resistance to these drugs.

  2. ZINC-INDUCED FACILITATOR-LIKE family in plants: lineage-specific expansion in monocotyledons and conserved genomic and expression features among rice (Oryza sativa paralogs

    Directory of Open Access Journals (Sweden)

    Lopes Karina L

    2011-01-01

    Full Text Available Abstract Background Duplications are very common in the evolution of plant genomes, explaining the high number of members in plant gene families. New genes born after duplication can undergo pseudogenization, neofunctionalization or subfunctionalization. Rice is a model for functional genomics research, an important crop for human nutrition and a target for biofortification. Increased zinc and iron content in the rice grain could be achieved by manipulation of metal transporters. Here, we describe the ZINC-INDUCED FACILITATOR-LIKE (ZIFL gene family in plants, and characterize the genomic structure and expression of rice paralogs, which are highly affected by segmental duplication. Results Sequences of sixty-eight ZIFL genes, from nine plant species, were comparatively analyzed. Although related to MSF_1 proteins, ZIFL protein sequences consistently grouped separately. Specific ZIFL sequence signatures were identified. Monocots harbor a larger number of ZIFL genes in their genomes than dicots, probably a result of a lineage-specific expansion. The rice ZIFL paralogs were named OsZIFL1 to OsZIFL13 and characterized. The genomic organization of the rice ZIFL genes seems to be highly influenced by segmental and tandem duplications and concerted evolution, as rice genome contains five highly similar ZIFL gene pairs. Most rice ZIFL promoters are enriched for the core sequence of the Fe-deficiency-related box IDE1. Gene expression analyses of different plant organs, growth stages and treatments, both from our qPCR data and from microarray databases, revealed that the duplicated ZIFL gene pairs are mostly co-expressed. Transcripts of OsZIFL4, OsZIFL5, OsZIFL7, and OsZIFL12 accumulate in response to Zn-excess and Fe-deficiency in roots, two stresses with partially overlapping responses. Conclusions We suggest that ZIFL genes have different evolutionary histories in monocot and dicot lineages. In rice, concerted evolution affected ZIFL duplicated genes

  3. RICE CROSSES

    African Journals Online (AJOL)

    Spikelet sterility is the greatest barrier to rice hybridization. ... sterility, Hybridization, Duplicate recessive epis- tasis ' ... hybrid fertility of intra-subspecific crosses, i.e., indica by ..... 'Germ 59(1) 293$ ' ' male sterility and fertility restoration in rice.

  4. Oncologic doses of zoledronic acid induce site specific suppression of bone modelling in rice rats.

    Science.gov (United States)

    Exposto, C R; Oz, U; Callard, J S; Allen, M J; Khurana, H; Atri, A D'; Mo, X; Fernandez, S A; Tatakis, D N; Edmonds, K; Westgate, P M; Huja, S S

    2017-06-01

    To examine the effect of zoledronic acid (ZOL) on cortical bone modelling and healing of extraction sockets in the jaw bones of a rodent model. We hypothesized ZOL suppresses both the bone formation in the modelling mode in the jaw bones and alters the extraction site healing. Rice rats were administered saline solution and two dose regimens of ZOL: 0.1 mg/kg, twice a week, for 4 weeks (n=17, saline=8 & ZOL=9) and a higher dose of 0.4 mg/kg, weekly, for 9 weeks (n=30, saline=15 & ZOL=15). Two pairs of fluorochrome bone labels were administered. Extraction of maxillary teeth was performed in maxilla. Mineral apposition rate, mineralizing surface and bone formation rate (BFR) were quantified on periodontal (PDL), alveolar and basal bone surfaces, and in the trabecular bone of proximal tibia. Bone volume (BV) was evaluated at extraction sockets. Multivariate Gaussian models were used to account for repeated measurements, and analyzes were conducted in SAS V9.3. ZOL suppressed bone modelling (BFR/BS) at the PDL surfaces in the mandible (Pmodelling mode in the jaws demonstrates the site specific effects of ZOL in rice rats. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  5. Comprehensive gene expression analysis of rice aleurone cells: probing the existence of an alternative gibberellin receptor.

    Science.gov (United States)

    Yano, Kenji; Aya, Koichiro; Hirano, Ko; Ordonio, Reynante Lacsamana; Ueguchi-Tanaka, Miyako; Matsuoka, Makoto

    2015-02-01

    Current gibberellin (GA) research indicates that GA must be perceived in plant nuclei by its cognate receptor, GIBBERELLIN INSENSITIVE DWARF1 (GID1). Recognition of GA by GID1 relieves the repression mediated by the DELLA protein, a model known as the GID1-DELLA GA perception system. There have been reports of potential GA-binding proteins in the plasma membrane that perceive GA and induce α-amylase expression in cereal aleurone cells, which is mechanistically different from the GID1-DELLA system. Therefore, we examined the expression of the rice (Oryza sativa) α-amylase genes in rice mutants impaired in the GA receptor (gid1) and the DELLA repressor (slender rice1; slr1) and confirmed their lack of response to GA in gid1 mutants and constitutive expression in slr1 mutants. We also examined the expression of GA-regulated genes by genome-wide microarray and quantitative reverse transcription-polymerase chain reaction analyses and confirmed that all GA-regulated genes are modulated by the GID1-DELLA system. Furthermore, we studied the regulatory network involved in GA signaling by using a set of mutants defective in genes involved in GA perception and gene expression, namely gid1, slr1, gid2 (a GA-related F-box protein mutant), and gamyb (a GA-related trans-acting factor mutant). Almost all GA up-regulated genes were regulated by the four named GA-signaling components. On the other hand, GA down-regulated genes showed different expression patterns with respect to GID2 and GAMYB (e.g. a considerable number of genes are not controlled by GAMYB or GID2 and GAMYB). Based on these observations, we present a comprehensive discussion of the intricate network of GA-regulated genes in rice aleurone cells.

  6. Mitochondria-targeted molecules MitoQ and SS31 reduce mutant huntingtin-induced mitochondrial toxicity and synaptic damage in Huntington's disease.

    Science.gov (United States)

    Yin, Xiangling; Manczak, Maria; Reddy, P Hemachandra

    2016-05-01

    The objective of this study was to determine the protective effects of the mitochondria-targeted molecules MitoQ and SS31 in striatal neurons that stably express mutant huntingtin (Htt) (STHDhQ111/Q111) in Huntington's disease (HD). We studied mitochondrial and synaptic activities by measuring mRNA and the protein levels of mitochondrial and synaptic genes, mitochondrial function, and ultra-structural changes in MitoQ- and SS31-treated mutant Htt neurons relative to untreated mutant Htt neurons. We used gene expression analysis, biochemical methods, transmission electron microscopy (TEM) and confocal microscopy methods. In the MitoQ- and SS31-treated mutant Htt neurons, fission genes Drp1 and Fis1 were down-regulated, and fusion genes Mfn1, Mfn2 and Opa1 were up-regulated relative to untreated neurons, suggesting that mitochondria-targeted molecules reduce fission activity. Interestingly, the mitochondrial biogenesis genes PGC1α, PGC1β, Nrf1, Nrf2 and TFAM were up-regulated in MitoQ- and SS31-treated mutant Htt neurons. The synaptic genes synaptophysin and PSD95 were up-regulated, and mitochondrial function was normal in the MitoQ- and SS31-treated mutant Htt neurons. Immunoblotting findings of mitochondrial and synaptic proteins agreed with the mRNA findings. TEM studies revealed decreased numbers of structurally intact mitochondria in MitoQ- and SS31-treated mutant Htt neurons. These findings suggest that mitochondria-targeted molecules MitoQ and SS31 are protective against mutant Htt-induced mitochondrial and synaptic damage in HD neurons, and these mitochondria-targeted molecules are potential therapeutic molecules for the treatment of HD neurons. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  7. OsNPR1 negatively regulates herbivore-induced JA and ethylene signaling and plant resistance to a chewing herbivore in rice.

    Science.gov (United States)

    Li, Ran; Afsheen, Sumera; Xin, Zhaojun; Han, Xiu; Lou, Yonggen

    2013-03-01

    NPR1 (a non-expressor of pathogenesis-related genes1) has been reported to play an important role in plant defense by regulating signaling pathways. However, little to nothing is known about its function in herbivore-induced defense in monocot plants. Here, using suppressive substrate hybridization, we identified a NPR1 gene from rice, OsNPR1, and found that its expression levels were upregulated in response to infestation by the rice striped stem borer (SSB) Chilo suppressalis and rice leaf folder (LF) Cnaphalocrocis medinalis, and to mechanical wounding and treatment with jasmonic acid (JA) and salicylic acid (SA). Moreover, mechanical wounding induced the expression of OsNPR1 quickly, whereas herbivore infestation induced the gene more slowly. The antisense expression of OsNPR1 (as-npr1), which reduced the expression of the gene by 50%, increased elicited levels of JA and ethylene (ET) as well as of expression of a lipoxygenase gene OsHI-LOX and an ACC synthase gene OsACS2. The enhanced JA and ET signaling in as-npr1 plants increased the levels of herbivore-induced trypsin proteinase inhibitors (TrypPIs) and volatiles, and reduced the performance of SSB. Our results suggest that OsNPR1 is an early responding gene in herbivore-induced defense and that plants can use it to activate a specific and appropriate defense response against invaders by modulating signaling pathways.

  8. Brucella Rough Mutant Induce Macrophage Death via Activating IRE1α Pathway of Endoplasmic Reticulum Stress by Enhanced T4SS Secretion

    Directory of Open Access Journals (Sweden)

    Peng Li

    2017-09-01

    Full Text Available Brucella is a Gram-negative facultative intracellular pathogen that causes the worldwide zoonosis, known as brucellosis. Brucella virulence relies mostly on its ability to invade and replicate within phagocytic cells. The type IV secretion system (T4SS and lipopolysaccharide are two major Brucella virulence factors. Brucella rough mutants reportedly induce the death of infected macrophages, which is T4SS dependent. However, the underlying molecular mechanism remains unclear. In this study, the T4SS secretion capacities of Brucella rough mutant and its smooth wild-type strain were comparatively investigated, by constructing the firefly luciferase fused T4SS effector, BPE123 and VceC. In addition, quantitative real-time PCR and western blotting were used to analyze the T4SS expression. The results showed that T4SS expression and secretion were enhanced significantly in the Brucella rough mutant. We also found that the activity of the T4SS virB operon promoter was notably increased in the Brucella rough mutant, which depends on quorum sensing-related regulators of VjbR upregulation. Cell infection and cell death assays revealed that deletion of vjbR in the Brucella rough mutant absolutely abolished cytotoxicity within macrophages by downregulating T4SS expression. This suggests that up-regulation of T4SS promoted by VjbR in rough mutant ΔrfbE contribute to macrophage death. In addition, we found that the Brucella rough mutant induce macrophage death via activating IRE1α pathway of endoplasmic reticulum stress. Taken together, our study provide evidence that in comparison to the Brucella smooth wild-type strain, VjbR upregulation in the Brucella rough mutant increases transcription of the virB operon, resulting in overexpression of the T4SS gene, accompanied by the over-secretion of effecter proteins, thereby causing the death of infected macrophages via activating IRE1α pathway of endoplasmic reticulum stress, suggesting novel insights into the

  9. Heme oxygenase-1 delays gibberellin-induced programmed cell death of rice aleurone layers subjected to drought stress by interacting with nitric oxide

    Directory of Open Access Journals (Sweden)

    Huangming eWu

    2016-01-01

    Full Text Available Cereal aleurone layers undergo a gibberellin (GA-regulated process of programmed cell death (PCD following germination. Heme oxygenase-1 (HO-1 is known as a rate-liming enzyme in the degradation of heme to biliverdin IXα (BV, carbon monoxide (CO, and free iron ions (Fe2+. It is a critical component in plant development and adaptation to environment stresses. Our previous studies confirmed that HO-1 inducer hematin (Ht promotes the germination of rice seeds in drought (20% polyethylene glycol-6000, PEG conditions, but the corresponding effects of HO-1 on the alleviation of germination-triggered PCD in GA-treated rice aleurone layers remain unknown. The present study has determined that GA co-treated with PEG results in lower HO-1 transcript levels and HO activity, which in turn results in the development of vacuoles in aleurone cells, followed by PCD. The pharmacology approach illustrated that up- or down-regulated HO-1 gene expression and HO activity delayed or accelerated GA-induced PCD. Furthermore, the application of the HO-1 inducer hematin and nitric oxide (NO donor sodium nitroprusside (SNP not only activated HO-1 gene expression, HO activity, and endogenous NO content, but also blocked GA-induced rapid vacuolation and accelerated aleurone layers PCD under drought stress. However, both HO-1 inhibitor zinc protoporphyrin IX (ZnPPIX and NO scavenger 2-(4-carboxyphenyl0-4, 4, 5, 5-tetramethylimidazoline-l-oxyl-3-oxide potassium salt (cPTIO reserved the effects of hematin and SNP on rice aleurone layer PCD under drought stress by down-regulating endogenous HO-1 and NO, respectively. The inducible effects of hematin and SNP on HO-1 gene expression, HO activity, and NO content were blocked by cPTIO. Together, these results clearly suggest that HO-1 is involved in the alleviation of GA-induced PCD of drought-triggered rice aleurone layers by associating with NO.

  10. The C2H2-type Zinc Finger Protein ZFP182 is Involved in Abscisic Acid-Induced Antioxidant Defense in Rice

    Institute of Scientific and Technical Information of China (English)

    Hong Zhang; Lan Ni; Yanpei Liu; Yunfei Wang; Aying Zhang; Mingpu Tan; Mingyi Jiang

    2012-01-01

    C2H2-type zinc finger proteins (ZFPs) are thought to play important roles in modulating the responses of plants to drought,salinity and oxidative stress.However,direct evidence is lacking for the involvement of these ZFPs in abscisic acid (ABA)-induced antioxidant defense in plants.In this study,the role of the rice (Oryza sativa L.sub.japonica cv.Nipponbare) C2H2-type ZFP ZFP182 in ABA-induced antioxidant defense and the relationship between ZFP182 and two rice MAPKs,OsMPK1 and OsMPK5 in ABA signaling were investigated.ABA treatment induced the increases in the expression of ZFP182,OsMPK1 and OsMPK5,and the activities of superoxide dismutase (SOD) and ascorbate peroxidase (APX) in rice leaves.The transient gene expression analysis and the transient RNA interference (RNAi) analysis in protoplasts showed that ZFP182,OsMPK1 and OsMPK5 are involved in ABA-induced up-regulation in the activities of SOD and APX.Besides,OsMPK1 and OsMPK5 were shown to be required for the up-regulation in the expression of ZFP182 in ABA signaling,but ZFP182 did not mediate the ABA-induced up-regulation in the expression of OsMPK1 and OsMPK5.These results indicate that ZFP182 is required for ABA-induced antioxidant defense and the expression of ZFP182 is regulated by rice MAPKs in ABA signaling.

  11. Heme Oxygenase-1 Delays Gibberellin-Induced Programmed Cell Death of Rice Aleurone Layers Subjected to Drought Stress by Interacting with Nitric Oxide.

    Science.gov (United States)

    Wu, Huangming; Zheng, Yan; Liu, Jing; Zhang, Heting; Chen, Huiping

    2015-01-01

    Cereal aleurone layers undergo a gibberellin (GA)-regulated process of programmed cell death (PCD) following germination. Heme oxygenase-1 (HO-1) is known as a rate-liming enzyme in the degradation of heme to biliverdin IXα, carbon monoxide (CO), and free iron ions (Fe(2+)). It is a critical component in plant development and adaptation to environment stresses. Our previous studies confirmed that HO-1 inducer hematin (Ht) promotes the germination of rice seeds in drought (20% polyethylene glycol-6000, PEG) conditions, but the corresponding effects of HO-1 on the alleviation of germination-triggered PCD in GA-treated rice aleurone layers remain unknown. The present study has determined that GA co-treated with PEG results in lower HO-1 transcript levels and HO activity, which in turn results in the development of vacuoles in aleurone cells, followed by PCD. The pharmacology approach illustrated that up- or down-regulated HO-1 gene expression and HO activity delayed or accelerated GA-induced PCD. Furthermore, the application of the HO-1 inducer Ht and nitric oxide (NO) donor sodium nitroprusside (SNP) not only activated HO-1 gene expression, HO activity, and endogenous NO content, but also blocked GA-induced rapid vacuolation and accelerated aleurone layers PCD under drought stress. However, both HO-1 inhibitor zinc protoporphyrin IX (ZnPPIX) and NO scavenger 2-(4-carboxyphenyl0-4, 4,5,5-tetramethylimidazoline-l-oxyl-3-oxide potassium salt (cPTIO) reserved the effects of Ht and SNP on rice aleurone layer PCD under drought stress by down-regulating endogenous HO-1 and NO, respectively. The inducible effects of Ht and SNP on HO-1 gene expression, HO activity, and NO content were blocked by cPTIO. Together, these results clearly suggest that HO-1 is involved in the alleviation of GA-induced PCD of drought-triggered rice aleurone layers by associating with NO.

  12. Stress-inducible expression of AtDREB1A transcription factor greatly improves drought stress tolerance in transgenic indica rice.

    Science.gov (United States)

    Ravikumar, G; Manimaran, P; Voleti, S R; Subrahmanyam, D; Sundaram, R M; Bansal, K C; Viraktamath, B C; Balachandran, S M

    2014-06-01

    The cultivation of rice (Oryza sativa L.), a major food crop, requires ample water (30 % of the fresh water available worldwide), and its productivity is greatly affected by drought, the most significant environmental factor. Much research has focussed on identifying quantitative trait loci, stress-regulated genes and transcription factors that will contribute towards the development of climate-resilient/tolerant crop plants in general and rice in particular. The transcription factor DREB1A, identified from the model plant Arabidopsis thaliana, has been reported to enhance stress tolerance against drought stress. We developed transgenic rice plants with AtDREB1A in the background of indica rice cultivar Samba Mahsuri through Agrobacterium-mediated transformation. The AtDREB1A gene was stably inherited and expressed in T1 and T2 plants and in subsequent generations, as indicated by the results of PCR, Southern blot and RT-PCR analyses. Expression of AtDREB1A was induced by drought stress in transgenic rice lines, which were highly tolerant to severe water deficit stress in both the vegetative and reproductive stages without affecting their morphological or agronomic traits. The physiological studies revealed that the expression of AtDREB1A was associated with an increased accumulation of the osmotic substance proline, maintenance of chlorophyll, increased relative water content and decreased ion leakage under drought stress. Most of the homozygous lines were highly tolerant to drought stress and showed significantly a higher grain yield and spikelet fertility relative to the nontransgenic control plants under both stressed and unstressed conditions. The improvement in drought stress tolerance in combination with agronomic traits is very essential in high premium indica rice cultivars, such as Samba Mahsuri, so that farmers can benefit in times of seasonal droughts and water scarcity.

  13. Effects of exogenous plant growth regulator abscisic acid-induced resistance in rice on the expression of vitellogenin mRNA in Nilaparvata lugens (Hemiptera: Delphacidae) adult females.

    Science.gov (United States)

    Liu, Jing-Lan; Chen, Xiao; Zhang, Hong-Mei; Yang, Xia; Wong, Andrew

    2014-01-01

    Recent study showed that exogenous abscisic acid (ABA) acts as a regulator of plant resistance. This study investigated average injury scale and callose contents of rice, and vitellogenin (Nlvg) mRNA expression in Nilaparvata lugens (Stål) (Hemiptera: Delphacidae) adult females after third instar nymphs fed on exogenous ABA-treated susceptible [Taichung Native one (TN1)] and moderately resistant (IR42) rice cultivars. The results showed that exogenous ABA significantly decreased average injury scale of rice and Nlvg mRNA expression in N. lugens adults compared with the control (without ABA spraying). Nlvg mRNA expression in N. lugens adults decreased significantly after third instar nymphs fed on ABA-treated (5, 20, and 40 mg/liter) TN1 for 1 and 2 d, and for IR42, after fed on ABA-treated (20 and 40 mg/liter) rice plants for 1 d and after fed on ABA-treated (5, 20, and 40 mg/liter) rice for 2 d decreased significantly. The callose contents showed no significant change for TN1, while for IR42, significantly increased in roots and sheathes after N. lugens infestation under ABA treatments (20 and 40 mg/liter) compared with the control. The decrease of Nlvg mRNA expression may be partially attributed to the increase of callose content of plants. The results provide a profile for concerning the effects of ABA-induced rice plants' defenses on phloem-feeding insects.

  14. Laser Irradiation-Induced DNA Methylation Changes Are Heritable and Accompanied with Transpositional Activation of mPing in Rice

    Science.gov (United States)

    Li, Siyuan; Xia, Qiong; Wang, Fang; Yu, Xiaoming; Ma, Jian; Kou, Hongping; Lin, Xiuyun; Gao, Xiang; Liu, Bao

    2017-01-01

    DNA methylation is an integral component of the epigenetic code in most higher eukaryotes. Exploring the extent to which DNA methylation can be altered under a specific condition and its heritability is important for elucidating the biological functions of this epigenetic modification. Here, we conducted MSAP analysis of rice plants with altered phenotypes subsequent to a low-dose Nd3+YAG laser irradiation. We found that all four methylation patterns at the 5′-CCGG sites that are analyzable by MSAP showed substantial changes in the immediately treated M0 plants. Interestingly, the frequencies of hypo- and hypermethylation were of similar extents, which largely offset each other and render the total methylation levels unchanged. Further analysis revealed that the altered methylation patterns were meiotically heritable to at least the M2 generation but accompanied with further changes in each generation. The methylation changes and their heritability of the metastable epigenetic state were verified by bisulfite sequencing of portion of the retrotranspon, Tos17, an established locus for assessing DNA methylation liability in rice. Real-time PCR assay indicated that the expression of various methylation-related chromatin genes was perturbed, and a Pearson correlation analysis showed that many of these genes, especially two AGOs (AGO4-1 and AGO4-2), were significantly correlated with the methylation pattern alterations. In addition, excisions of a MITE transposon, mPing, occurred rampantly in the laser irradiated plants and their progenies. Together, our results indicate that heritable DNA methylation changes can be readily induced by low-dose laser irradiation, and which can be accompanied by transpostional activation of transposable elements.

  15. High Temperature Induced Glume Closure Resulted in Lower Fertility in Hybrid Rice Seed Production

    Science.gov (United States)

    Yan, Haoliang; Zhang, Binglin; Zhang, Yunbo; Chen, Xinlan; Xiong, Hui; Matsui, Tsutomu; Tian, Xiaohai

    2017-01-01

    Predicted climate changes, in particular, the increased dimension and frequency of heat waves, are expected to affect crop growth in the future seriously. Hybrid rice relies on seed production from male sterile and restorer lines. Experiments were conducted over two consecutive years to compare the high temperature tolerance of parents of different hybrid rice combinations, in terms of fertility rate, flowering pattern, pollination and physiological parameters of the lodicule. Three male sterile lines and a broad compatibility restorer line (as pollen donor and conventional variety as well) were grown to heading stage and then treated with average daily temperatures of 26°C (range 23–30°C), 28°C (25–32°C), and 30°C (26–34°C), respectively, continued for 5–7 days each in a natural light phytotron which simulated the local typical high temperature weather in the field. The results indicated that male sterile lines were more sensitive to high temperature than the restorer line for fertility rate, and the sensitivity varied between varieties. The fertility rate of the restorer line was maintained at about 90% under the high temperature treatments, while it decreased in the male sterile lines by 23.3 and 48.1% at 28 and 30°C, respectively. The fertility rate of the most sensitive line declined by 70%, and the tolerant line declined by 34% at 30°C. Glume closure in the male sterile lines was a major reason for the reduced fertility rate under high temperature, which is closely correlated with carbohydrates content and the vascular bundle pattern in the lodicule. The present study identified a useful trait to select male sterile lines with high temperature tolerance for seed production. PMID:28105031

  16. Chelator-induced phytoextraction of zinc and copper by rice seedlings.

    Science.gov (United States)

    Yu, Xiao-Zhang; Wang, Dun-Qiu; Zhang, Xue-Hong

    2014-05-01

    Solution culture was carried to investigate capacity of synthetic aminopolycarboxylic acids (ethylenediamine tetraacetate, N-hydroxyethylenediaminetriacetic acid, and diethylenetriamine-pentaacetate) for enhancing botanical removal and transport of heavy metals (Cu and Zn) by plants. Biodegradable organic acids (citric acid, malic acid, and oxalic acid) were also selected as alternatives to compare them with synthesized chelating agents for effectiveness. Young rice seedlings (Oryza sativa L. cv. XZX 45) were grown in nutrient solutions treated with single or combined metal solutions in presence or absence of chelating compounds. Calculation by chemical equilibrium program VISUAL MINTEQ showed that different chelating compounds had various complex potential with Cu(2+) and Zn(2+) ions, in which synthetic chelators exhibited higher complexed capability than biodegradable organic acids. All applied synthetic aminopolycarboxylic acids significantly decreased removal of metal from nutrient solution (p 0.05), compared with the treatment without metal ligands. Synthetic aminopolycarboxylic acids significantly decreased metal concentrations in plant materials in all treatments (p < 0.01). However, biodegradable organic acids decreased metal concentrations in roots (p < 0.01), but enhanced them in shoots (p < 0.01). Results obtained indicated that synthetic aminopolycarboxylic acids decreased uptake of metals by rice seedlings, but translocation of metals complexed within plant materials was evident. Although exogenous biodegradable organic acids showed negligible effect on botanical removal of metals, metals complexed with organic acids was more mobile than those complexed with other chelating agents. These information collected here had important implication for the use of biodegradable metal chelators in transport of essential micronutrients in plant nutrition.

  17. Research Progress of Induced Resistance against Rice Blast and Its Resistance Mechanisms%诱导水稻抗瘟性及诱抗机理研究进展

    Institute of Scientific and Technical Information of China (English)

    胡能; 涂国全; 黄林

    2015-01-01

    Rice blast is the most harmful rice disease among three major rice diseases. Induced resistance against rice blast will play an important role in the control of rice blast for its following characteristics:persistence, broad spectrum, safety, systemic,etc. Recent researches on induced resistance against rice blast, the biotic and abiotic induced resistance elicitors, and resistance mechanisms were introduced in this paper. Existing problems about the research and its development direction about induced resistance against rice blast in the future were also discussed.%稻瘟病是危害水稻三大病害之首,水稻稻瘟病诱导抗病性具有持久性、广谱性、安全性、系统性等而对未来水稻稻瘟病的防治发挥巨大的作用。综述近年来国内外水稻诱导抗瘟性的研究进展,对生物及非生物诱导因子和诱导机理的三个方面进行了介绍,并提出对现在一些研究不足以及未来的发展方向。

  18. Distinct Neurodegenerative Changes in an Induced Pluripotent Stem Cell Model of Frontotemporal Dementia Linked to Mutant TAU Protein

    Directory of Open Access Journals (Sweden)

    Marc Ehrlich

    2015-07-01

    Full Text Available Frontotemporal dementia (FTD is a frequent form of early-onset dementia and can be caused by mutations in MAPT encoding the microtubule-associated protein TAU. Because of limited availability of neural cells from patients’ brains, the underlying mechanisms of neurodegeneration in FTD are poorly understood. Here, we derived induced pluripotent stem cells (iPSCs from individuals with FTD-associated MAPT mutations and differentiated them into mature neurons. Patient iPSC-derived neurons demonstrated pronounced TAU pathology with increased fragmentation and phospho-TAU immunoreactivity, decreased neurite extension, and increased but reversible oxidative stress response to inhibition of mitochondrial respiration. Furthermore, FTD neurons showed an activation of the unfolded protein response, and a transcriptome analysis demonstrated distinct, disease-associated gene expression profiles. These findings indicate distinct neurodegenerative changes in FTD caused by mutant TAU and highlight the unique opportunity to use neurons differentiated from patient-specific iPSCs to identify potential targets for drug screening purposes and therapeutic intervention.

  19. Changes in DNA base sequences in the mutant of Arabidopsis thaliana induced by low-energy N+ implantation

    Institute of Scientific and Technical Information of China (English)

    常凤启; 刘选明; 李银心; 贾庚祥; 马晶晶; 刘公社; 朱至清

    2003-01-01

    To reveal the mutation effect of low-energy ion implantation on Arabidopsis thaliana in vivo, T80II, a stable dwarf mutant, derived from the seeds irradiated by 30 keV N+ with the dose of 80×1015 ions/cm2 was used for Random Amplified Polymorphic DNA (RAPD) and base sequence analysis. The results indicated that among total 397 RAPD bands observed, 52 bands in T80II were different from those of wild type showing a variation frequency 13.1%. In comparison with the sequences of A. thaliana in GenBank, the RAPD fragments in T80II were changed greatly in base sequences with an average rate of one base change per 16.8 bases. The types of base changes included base transition, transversion, deletion and insertion. Among the 275 base changes detected, single base substitutions (97.09%) occurred more frequently than base deletions and insertions (2.91%). And the frequency of base transitions (66.55%) was higher than that of base transversions (30.55%). Adenine, thymine, guanine or cytosine could be replaced by any of other three bases in cloned DNA fragments in T80II. It seems that thymine was more sensitive to the irradiation than other bases. The flanking sequences of the base changes in RAPD fragments in T80II were analyzed and the mutational "hotspot" induced by low-energy ion implantation was discussed.

  20. Genetically Engineered Ascorbic acid-deficient Live Mutants of Leishmania donovani induce long lasting Protective Immunity against Visceral Leishmaniasis.

    Science.gov (United States)

    Anand, Sneha; Madhubala, Rentala

    2015-06-02

    Visceral leishmaniasis caused by Leishmania donovani is the most severe systemic form of the disease. There are still no vaccines available for humans and there are limitations associated with the current therapeutic regimens for leishmaniasis. Recently, we reported functional importance of Arabino-1, 4-lactone oxidase (ALO) enzyme from L. donovani involved in ascorbate biosynthesis pathway. In this study, we have shown that ΔALO parasites do not affect the ability of null mutants to invade visceral organs but severely impair parasite persistence beyond 16 week in BALB/c mice and hence are safe as an immunogen. Both short term (5 week) and long term (20 week) immunization with ΔALO parasites conferred sustained protection against virulent challenge in BALB/c mice, activated splenocytes and resulted in induction of pro-inflammatory cytokine response. Protection in immunized mice after challenge correlated with the stimulation of IFN-γ producing CD4(+) and CD8(+) T cells. Antigen-mediated cell immunity correlated with robust nitrite and superoxide generation, macrophage-derived oxidants critical in controlling Leishmania infection. Our data shows that live attenuated ΔALO parasites are safe, induce protective immunity and can provide sustained protection against Leishmania donovani. We further conclude that the parasites attenuated in their anti-oxidative defence mechanism can be exploited as vaccine candidates.

  1. Increased lipolysis and altered lipid homeostasis protect γ-synuclein–null mutant mice from diet-induced obesity

    Science.gov (United States)

    Millership, Steven; Ninkina, Natalia; Guschina, Irina A.; Norton, Jessica; Brambilla, Riccardo; Oort, Pieter J.; Adams, Sean H.; Dennis, Rowena J.; Voshol, Peter J.; Rochford, Justin J.; Buchman, Vladimir L.

    2012-01-01

    Synucleins are a family of homologous proteins principally known for their involvement in neurodegeneration. γ-Synuclein is highly expressed in human white adipose tissue and increased in obesity. Here we show that γ-synuclein is nutritionally regulated in white adipose tissue whereas its loss partially protects mice from high-fat diet (HFD)–induced obesity and ameliorates some of the associated metabolic complications. Compared with HFD-fed WT mice, HFD-fed γ-synuclein–null mutant mice display increased lipolysis, lipid oxidation, and energy expenditure, and reduced adipocyte hypertrophy. Knockdown of γ-synuclein in adipocytes causes redistribution of the key lipolytic enzyme ATGL to lipid droplets and increases lipolysis. γ-Synuclein–deficient adipocytes also contain fewer SNARE complexes of a type involved in lipid droplet fusion. We hypothesize that γ-synuclein may deliver SNAP-23 to the SNARE complexes under lipogenic conditions. Via these independent but complementary roles, γ-synuclein may coordinately modulate lipid storage by influencing lipolysis and lipid droplet formation. Our data reveal γ-synuclein as a regulator of lipid handling in adipocytes, the function of which is particularly important in conditions of nutrient excess. PMID:23213245

  2. Increased lipolysis and altered lipid homeostasis protect γ-synuclein-null mutant mice from diet-induced obesity.

    Science.gov (United States)

    Millership, Steven; Ninkina, Natalia; Guschina, Irina A; Norton, Jessica; Brambilla, Riccardo; Brambilla, Ricardo; Oort, Pieter J; Adams, Sean H; Dennis, Rowena J; Voshol, Peter J; Rochford, Justin J; Buchman, Vladimir L

    2012-12-18

    Synucleins are a family of homologous proteins principally known for their involvement in neurodegeneration. γ-Synuclein is highly expressed in human white adipose tissue and increased in obesity. Here we show that γ-synuclein is nutritionally regulated in white adipose tissue whereas its loss partially protects mice from high-fat diet (HFD)-induced obesity and ameliorates some of the associated metabolic complications. Compared with HFD-fed WT mice, HFD-fed γ-synuclein-null mutant mice display increased lipolysis, lipid oxidation, and energy expenditure, and reduced adipocyte hypertrophy. Knockdown of γ-synuclein in adipocytes causes redistribution of the key lipolytic enzyme ATGL to lipid droplets and increases lipolysis. γ-Synuclein-deficient adipocytes also contain fewer SNARE complexes of a type involved in lipid droplet fusion. We hypothesize that γ-synuclein may deliver SNAP-23 to the SNARE complexes under lipogenic conditions. Via these independent but complementary roles, γ-synuclein may coordinately modulate lipid storage by influencing lipolysis and lipid droplet formation. Our data reveal γ-synuclein as a regulator of lipid handling in adipocytes, the function of which is particularly important in conditions of nutrient excess.

  3. Apc-Mutant Kyoto Apc Delta (KAD) Rats Are Susceptible to 4-NQO-Induced Tongue Carcinogenesis

    Energy Technology Data Exchange (ETDEWEB)

    Tanaka, Takuji, E-mail: tmntt08@gmail.com [Department of Diagnostic Pathology (DDP) & Research Center of Diagnostic Pathology (RC-DiP), Gifu Municipal Hospital, 7-1 Kashima-Cho, Gifu 500-8513 (Japan); Department of Tumor Pathology, Gifu University Graduate School of Medicine, 1-1 Yanagido, Gifu 501-1194 (Japan); Shimizu, Masahito; Kochi, Takahiro; Shirakami, Yohei [Department of Internal Medicine/Gastroenterology, Gifu University Graduate School of Medicine, 1-1 Yanagido, Gifu 501-1194 (Japan); Mori, Takayuki [Department of Pharmacy, Ogaki Municipal Hospital, 4-86 Minaminokawa-cho, Ogaki 503-8502 (Japan); Watanabe, Naoki [Department of Diagnostic Pathology (DDP) & Research Center of Diagnostic Pathology (RC-DiP), Gifu Municipal Hospital, 7-1 Kashima-Cho, Gifu 500-8513 (Japan); Naiki, Takafumi [Department of Clinical Laboratory, Gifu Municipal Hospital, 7-1 Kashima-cho, Gifu 500-8513 (Japan); Moriwaki, Hisataka [Department of Internal Medicine/Gastroenterology, Gifu University Graduate School of Medicine, 1-1 Yanagido, Gifu 501-1194 (Japan); Yoshimi, Kazuto; Serikawa, Tadao; Kuramoto, Takashi [The Institute of Laboratory Animals, Graduate School of Medicine, Kyoto University, Yoshidakonoe-cho, Sakyo-ku, Kyoto 606-8501 (Japan)

    2014-07-21

    Despite widening interest in the possible association between infection/inflammation and cancer development, knowledge of this issue in relation to oral cancer remains inadequate. This study aimed to determine the susceptibility of Apc-mutant Kyoto Apc Delta (KAD) rats, which are vulnerable to developing inflammation-associated colorectal carcinogenesis, to 4-nitroquinoline 1-oxide (4-NQO)-induced tongue carcinogenesis in order to clarify the role of inflammation in oral cancer. KAD (20 males and 22 females) and F344/NS1c (22 males and 23 females) rats received drinking water with or without 4-NQO (20 ppm) for eight weeks. Histopathological and immunohistochemical analyses of the tongue were performed at week 20. Additionally, the mRNA expression of inflammatory cytokines in the tongue mucosa was determined at week 8. Tongue squamous cell carcinoma (SCC) developed in the KAD and F344/NS1c rats that received 4-NQO. Regardless of gender, the incidence and multiplicity of tongue SCC were greater in the KAD rats than in the F344/NS1c rats. In addition, the multiplicity of tongue SCC in the female KAD rats was significantly greater than that observed in the male KAD (p < 0.01) and female F344/NS1c rats (p < 0.05). The levels of inflammation and the mRNA expression of inflammatory cytokines in the tongue in the 4-NQO-treated female KAD rats were the highest among the rats given 4-NQO. These results show that KAD rats, particularly females, are susceptible to 4-NQO-induced tongue carcinogenesis, suggesting the utility of models employing KAD rats for investigating the pathobiology of oral (tongue) carcinogenesis associated with inflammation.

  4. Tomato fruit development in the auxin-resistant dgt mutant is induced by pollination but not by auxin treatment.

    Science.gov (United States)

    Mignolli, Francesco; Mariotti, Lorenzo; Lombardi, Lara; Vidoz, María Laura; Ceccarelli, Nello; Picciarelli, Piero

    2012-08-15

    In tomato (Solanum lycopersicum Mill.), auxin is believed to play a pivotal role in controlling fruit-set and early ovary growth. In this paper we investigated the effect of the reduced auxin sensitivity exhibited by the diageotropica (dgt) tomato mutant on ovary growth during early stage of fruit development. Here we show that in hand-pollinated ovaries fruit-set was not affected by the dgt lesion while fruit growth was reduced. This reduction was associated with a smaller cell size of mesocarp cells, with a lower mean C values and with a lower gene expression of the expansin gene LeExp2. When a synthetic auxin (4-CPA, chlorophenoxyacetic acid) was applied to the flowers of wild type plants, parthenocarpic ovary growth was induced. On the contrary, auxin application to the flowers of dgt plants failed to induce parthenocarpy. Hand-pollinated ovaries of dgt contained higher levels of IAA compared to wild type and this was not associated with high transcript levels of genes encoding a key regulatory enzyme of IAA biosynthesis (ToFZYs) but with lower expression levels of GH3, a gene involved in the conjugation of IAA to amino acids. The expression of diverse Aux/IAA genes and SAUR (small auxin up-regulated RNA) was also altered in the dgt ovaries. The dgt lesion does not seem to affect specific Aux/IAA genes in terms of transcript occurrence but rather in terms of relative levels of expression. Transcript levels of Aux/IAA genes were up regulated in auxin-treated ovaries of wild-type but not in dgt. Together, our results suggest that dgt ovary cells are not able to sense and/or transduce the external auxin signal, whereas pollinated dgt ovary cells are able to detect the IAA present in fertilized ovules promoting fruit development.

  5. Apc-Mutant Kyoto Apc Delta (KAD Rats Are Susceptible to 4-NQO-Induced Tongue Carcinogenesis

    Directory of Open Access Journals (Sweden)

    Takuji Tanaka

    2014-07-01

    Full Text Available Despite widening interest in the possible association between infection/ inflammation and cancer development, knowledge of this issue in relation to oral cancer remains inadequate. This study aimed to determine the susceptibility of Apc-mutant Kyoto Apc Delta (KAD rats, which are vulnerable to developing inflammation-associated colorectal carcinogenesis, to 4-nitroquinoline 1-oxide (4-NQO-induced tongue carcinogenesis in order to clarify the role of inflammation in oral cancer. KAD (20 males and 22 females and F344/NS1c (22 males and 23 females rats received drinking water with or without 4-NQO (20 ppm for eight weeks. Histopathological and immunohistochemical analyses of the tongue were performed at week 20. Additionally, the mRNA expression of inflammatory cytokines in the tongue mucosa was determined at week 8. Tongue squamous cell carcinoma (SCC developed in the KAD and F344/NS1c rats that received 4-NQO. Regardless of gender, the incidence and multiplicity of tongue SCC were greater in the KAD rats than in the F344/NS1c rats. In addition, the multiplicity of tongue SCC in the female KAD rats was significantly greater than that observed in the male KAD (p < 0.01 and female F344/NS1c rats (p < 0.05. The levels of inflammation and the mRNA expression of inflammatory cytokines in the tongue in the 4-NQO-treated female KAD rats were the highest among the rats given 4-NQO. These results show that KAD rats, particularly females, are susceptible to 4-NQO-induced tongue carcinogenesis, suggesting the utility of models employing KAD rats for investigating the pathobiology of oral (tongue carcinogenesis associated with inflammation.

  6. 脯氨酸代谢途径在调控水稻phyB突变体干旱胁迫耐性中的作用%Involvement of Proline Metabolic Pathway in Regulating Drought Tolerance in Rice phyB Mutant

    Institute of Scientific and Technical Information of China (English)

    周晋军; 刘蓬; 尹静静; 谢先芝

    2014-01-01

    The expression levels of key genes involved in proline metabolic pathway were compared in rice wild type (WT)and phyB mutant.The results showed that dehydration treatment induced the expression of proline biosynthetic gene OsP5CS1 and OsOAT,but suppressed the expression of proline catabolic gene OsP5CDH.And the expression level of OsP5CS1 gene in phyB mutant was obviously higher than that in WT, which suggesting that phyB negatively regulated its expression.To explore the relation between high OsP5CS1 transcript level and stronger drought tolerance in phyB mutant,the OsP5CS1 transgenic tobacco was obtained. Water loss rate assays of detached leaves revealed that transgenic tobacco showed a slower water loss rate com-pared to non-transgenic tobacco.In addition,the leaf disk differentiation rate under high salinity of transgen-ic tobacco was higher than that of non -transgenic tobacco.These findings suggested that the regulation of phyB on proline metabolic pathway was one of the important factors contributing to stronger drought tolerance in phyB mutant.%本研究分析了水稻野生型和phyB突变体中脯氨酸代谢途径关键基因的表达水平。结果显示,干旱处理能够诱导脯氨酸生物合成相关基因OsP5CS1和OsOAT的表达,抑制脯氨酸生物降解基因OsP5CDH的表达。且phyB突变体中OsP5CS1基因的表达水平明显高于野生型,据此推测phyB负调控OsP5CS1基因的表达。为了分析OsP5CS1基因的高水平表达是否与phyB突变体较强的干旱胁迫耐性有关,本研究进一步培育了转OsP5CS1基因烟草。离体叶片失水速率结果表明,转基因烟草的失水速率小于野生型;盐胁迫条件下,转基因烟草的分化率明显高于野生型。综上所述,phyB对脯氨酸代谢途径的调控是phyB突变体具有较强干旱胁迫耐性的重要因素之一。

  7. Use of a chemically induced-colon carcinogenesis-prone Apc-mutant rat in a chemotherapeutic bioassay

    Directory of Open Access Journals (Sweden)

    Yoshimi Kazuto

    2012-10-01

    Full Text Available Abstract Background Chemotherapeutic bioassay for colorectal cancer (CRC with a rat model bearing chemically-induced CRCs plays an important role in the development of new anti-tumor drugs and regimens. Although several protocols to induce CRCs have been developed, the incidence and number of CRCs are not much enough for the efficient bioassay. Recently, we established the very efficient system to induce CRCs with a chemically induced-colon carcinogenesis-prone Apc-mutant rat, Kyoto Apc Delta (KAD rat. Here, we applied the KAD rat to the chemotherapeutic bioassay for CRC and showed the utility of the KAD rat. Methods The KAD rat has been developed by the ENU mutagenesis and carries a homozygous nonsense mutation in the Apc gene (S2523X. Male KAD rats were given a single subcutaneous injection of AOM (20 mg/kg body weight at 5 weeks of age. Starting at 1 week after the AOM injection, they were given 2% DSS in drinking water for 7 days. Tumor-bearing KAD rats were divided into experimental and control groups on the basis of the number of tumors observed by endoscopy at week 8. The 5-fluorouracil (5-FU was administrated intravenously a dose of 50 or 75 mg/kg weekly at week 9, 10, and 11. After one-week interval, the 5-FU was given again at week 13, 14, and 15. At week 16, animals were sacrificed and tumor number and volume were measured macroscopically and microscopically. Results In total 48 tumors were observed in 27 KAD rats with a 100% incidence at week 8. The maximum tolerated dose for the KAD rat was 50 mg/kg of 5-FU. Macroscopically, the number or volume of tumors in the 5-FU treated rats was not significantly different from the control. Microscopically, the number of adenocarcinoma in the 5-FU treated rats was not significantly different (p Conclusion The use of the AOM/DSS-treated tumor-bearing KAD rats could shorten the experimental period and reduce the number of animals examined in the chemotherapeutic bioassay. The

  8. Mutant INS-gene induced diabetes of youth: proinsulin cysteine residues impose dominant-negative inhibition on wild-type proinsulin transport.

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    Ming Liu

    Full Text Available Recently, a syndrome of Mutant INS-gene-induced Diabetes of Youth (MIDY, derived from one of 26 distinct mutations has been identified as a cause of insulin-deficient diabetes, resulting from expression of a misfolded mutant proinsulin protein in the endoplasmic reticulum (ER of insulin-producing pancreatic beta cells. Genetic deletion of one, two, or even three alleles encoding insulin in mice does not necessarily lead to diabetes. Yet MIDY patients are INS-gene heterozygotes; inheritance of even one MIDY allele, causes diabetes. Although a favored explanation for the onset of diabetes is that insurmountable ER stress and ER stress response from the mutant proinsulin causes a net loss of beta cells, in this report we present three surprising and interlinked discoveries. First, in the presence of MIDY mutants, an increased fraction of wild-type proinsulin becomes recruited into nonnative disulfide-linked protein complexes. Second, regardless of whether MIDY mutations result in the loss, or creation, of an extra unpaired cysteine within proinsulin, Cys residues in the mutant protein are nevertheless essential in causing intracellular entrapment of co-expressed wild-type proinsulin, blocking insulin production. Third, while each of the MIDY mutants induces ER stress and ER stress response; ER stress and ER stress response alone appear insufficient to account for blockade of wild-type proinsulin. While there is general agreement that ultimately, as diabetes progresses, a significant loss of beta cell mass occurs, the early events described herein precede cell death and loss of beta cell mass. We conclude that the molecular pathogenesis of MIDY is initiated by perturbation of the disulfide-coupled folding pathway of wild-type proinsulin.

  9. Rice genotype differences in tolerance of zinc-deficient soils: evidence for the importance of root-induced changes in the rhizosphere

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    Asako eMori

    2016-01-01

    Full Text Available Zinc (Zn deficiency is a major constraint to rice production and Zn is also often deficient in humans with rice-based diets. Efforts to breed more Zn-efficient rice are constrained by poor understanding of the mechanisms of tolerance to deficiency. Here we assess the contributions of root growth and root Zn uptake efficiency, and we seek to explain the results in terms of specific mechanisms. We made a field experiment in a highly Zn-deficient rice soil in the Philippines with deficiency-tolerant and -sensitive genotypes, and measured growth, Zn uptake and root development. We also measured the effect of planting density. Tolerant genotypes produced more crown roots per plant and had greater uptake rates per unit root surface area; the latter was at least as important as root number to overall tolerance. Tolerant and sensitive genotypes took up more Zn per plant at greater planting densities. The greater uptake per unit root surface area, and the planting density effect can only be explained by root-induced changes in the rhizosphere, either solubilizing Zn or neutralizing a toxin that impedes Zn uptake (possibly HCO3- or Fe2+, or both. Traits for these and crown root number are potential breeding targets.

  10. Analysis of Magnaporthe oryzae genome reveals a fungal effector, which is able to induce resistance response in transgenic rice line containing resistance gene, Pi54

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    Soham Ray

    2016-08-01

    Full Text Available Rice blast caused by Magnaporthe oryzae is one of the most important diseases of rice. Pi54, a rice gene that imparts resistance to M. oryzae isolates prevalent in India, was already cloned but its avirulent counterpart in the pathogen was not known.. After decoding the whole genome of an avirulent isolate of M. oryzae, we predicted 11440 protein coding genes and then identified four candidate effector proteins which are exclusively expressed in the infectious structure, appresoria. In silico protein modeling followed by interaction analysis between Pi54 protein model and selected four candidate effector proteins models revealed that Mo-01947_9 protein model encoded by a gene located at chromosome 4 of M. oryzae, interacted best at the Leucine Rich Repeat domain of Pi54 protein model. Yeast-two-hybrid analysis showed that Mo-01947_9 protein physically interacts with Pi54 protein. Nicotiana benthamiana leaf infiltration assay confirmed induction of hypersensitive response in the presence of Pi54 gene in a heterologous system. Genetic complementation test also proved that Mo-01947_9 protein induces avirulence response in the pathogen in the presence of Pi54 gene. Here, we report identification and cloning of a new fungal effector gene which interacts with resistance gene Pi54 gene in rice.

  11. Analysis of transcriptional and upstream regulatory sequence activity of two environmental stress-inducible genes, NBS-Str1 and BLEC-Str8, of rice.

    Science.gov (United States)

    Ray, Swatismita; Kapoor, Sanjay; Tyagi, Akhilesh K

    2012-04-01

    Two abiotic stress-inducible upstream regulatory sequences (URSs) from rice have been identified and functionally characterized in rice. NBS-Str1 and BLEC-Str8 genes have been identified, by analysing the transcriptome data of cold, salt and desiccation stress-treated 7-day-old rice (Oryza sativa L. var. IR64) seedling, to be preferentially responsive to desiccation and salt stress, respectively. NBS-Str1 and BLEC-Str8 genes code for putative NBS (nucleotide binding site)-LRR (leucine rich repeat) and β-lectin domain protein, respectively. NBS-Str1 URS is induced in root tissue, preferentially in vascular bundle, during 3 and 24 h of desiccation stress condition in transgenic 7-day-old rice seedling. In mature transgenic plants, this URS shows induction in root and shoot tissue under desiccation stress as well as under prolonged (1 and 2 day) salt stress. BLEC-Str8 URS shows basal activity under un-stressed condition, however, it is inducible under salt stress condition in both root and leaf tissues in young seedling and mature plants. Activity of BLEC-Str8 URS has been found to be vascular tissue preferential, however, under salt stress condition its activity is also found in the mesophyll tissue. NBS-Str1 and BLEC-Str8 URSs are inducible by heavy metal, copper and manganese. Interestingly, both the URSs have been found to be non responsive to ABA treatment, implying them to be part of ABA-independent abiotic stress response pathway. These URSs could prove useful for expressing a transgene in a stress responsive manner for development of stress tolerant transgenic systems.

  12. Isolation of Rhizobium phaseoli Tn5-induced mutants with altered expression of cytochrome terminal oxidases o and aa3.

    Science.gov (United States)

    Soberón, M; Membrillo-Hernández, J; Aguilar, G R; Sánchez, F

    1990-01-01

    Two Rhizobium phaseoli mutants affected in cytochrome expression were obtained by Tn5-mob mutagenesis of the wild-type strain (CE3). Mutant strain CFN031 expressed sevenfold less cytochrome o in culture, expressed cytochrome aa3 under microaerophilic culture conditions, in contrast to strain CE3, and was affected in its vegetative growth properties and proliferation inside plant host cells. Mutant CFN037 expressed cytochrome aa3 under microaerophilic culture conditions, while bacteroid development and nitrogen fixation occurred earlier than in strain CE3. Images FIG. 2 PMID:2155209

  13. Free radical induced grafting of acrylonitrile on pre-treated rice straw for enhancing its durability and flame retardancy.

    Science.gov (United States)

    Mukherjee, Aparna; Halder, Seema; Datta, Deepshikha; Anupam, Kumar; Hazra, Biren; Kanti Mandal, Mrinal; Halder, Gopinath

    2017-01-01

    The present investigation highlights the feasibility of a polymer grafting process to enhance the durability and flame retardancy of rice straw towards application as a low cost roofing material. The success of this grafting methodology was perceived to depend upon a bi-step pre-treatment process encompassing delignification and inorganic salts dispersion. Subsequently free radical polymer grafting of acrylonitrile onto rice straw was implemented by immersion mechanism initiated by oxalic acid-potassium permanganate initiator. The percentage of grafting, limiting oxygen index (LOI), biodegradability of the grafted rice straw and grafting yield percentage was estimated to be 57%, 27%, 0.02% and 136.67%, respectively. The weight loss of polymer grafted rice straw implied its less biodegradability over raw straw. Thus, the process of grafting contrived in the present analysis can be a promising and reliable technique for the efficient utilization of rice straw as an inexpensive roofing element through the augmentation of its durability and flame retardancy.

  14. Knockdown of Cs-Spook induces delayed larval molting in rice striped stem borer Chilo suppressalis.

    Science.gov (United States)

    Shahzad, Muhammad Faisal; Li, Yao; Ge, Chang; Sun, Yang; Yang, Qiupu; Li, Fei

    2015-03-01

    Spook has essential roles in the biogenesis of the molting hormone 20-hydroxyecdysone (20-E). The function of spook in the rice striped stem borer (SSB) Chilo suppressalis remains unclear, prompting our hypothesis that it exerts actions similar to those reported for other insect species. Here we amplified the full-length transcript of spook (Cs-Spook) in SSB by 5' and 3' rapid amplification of cDNA ends. Cs-Spook has conserved P450 motifs such as Helix-C, Helix-I, Helix-K, and PERF motif (PxxFxPxRF). It was highly expressed in late instar larvae but less so in newly molted larvae. Cs-Spook was highly expressed in prothoracic glands. Cs-Spook was knocked down by dsRNA treatments. Compared with controls, the gene expression level was reduced to 9% at 24 h post injection (PI), 33% at 48 h PI, and 24% at 72 h PI. The ecdysteroid titer decreased significantly in the dsRNA-treated group (P < 0.05), resulting in delayed larval development. The delayed development in dsRNA-treatment group was rescued by treating with 20-E. Our work demonstrates that Cs-Spook participates in the biogenesis of 20-E and regulates the molt of SSB, as seen in other species.

  15. Drought stress-induced compositional changes in tolerant transgenic rice and its wild type.

    Science.gov (United States)

    Nam, Kyong-Hee; Kim, Do-Young; Shin, Hee Jae; Nam, Ki Jung; An, Joo Hee; Pack, In-Soon; Park, Jung-Ho; Jeong, Soon-Chun; Kim, Ho Bang; Kim, Chang-Gi

    2014-06-15

    Comparing well-watered versus deficit conditions, we evaluated the chemical composition of grains harvested from wild-type (WT) and drought-tolerant, transgenic rice (Oryza sativa L.). The latter had been developed by inserting AtCYP78A7, which encodes a cytochrome P450 protein. Two transgenic Lines, '10B-5' and '18A-4', and the 'Hwayoung' WT were grown under a rainout shelter. After the harvested grains were polished, their levels of key components, including proximates, amino acids, fatty acids, minerals and vitamins were analysed to determine the effect of watering system and genotype. Drought treatment significantly influenced the levels of some nutritional components in both transgenic and WT grains. In particular, the amounts of lignoceric acid and copper in the WT decreased by 12.6% and 39.5%, respectively, by drought stress, whereas those of copper and potassium in the transgenics rose by 88.1-113.3% and 10.4-11.9%, respectively, under water-deficit conditions.

  16. Exposure to tebuconazol in rice field and laboratory conditions induces oxidative stress in carp (Cyprinus carpio).

    Science.gov (United States)

    Toni, Cândida; Loro, Vania Lucia; Santi, Adriana; de Menezes, Charlene Cavalheiro; Cattaneo, Roberta; Clasen, Bárbara Estevão; Zanella, Renato

    2011-01-01

    Pesticides can have an effect on the biochemical and physiological functions of living organisms. The changes seen in fish and their response to pesticides can be used as an example for vertebrate toxicity. In this study, carp fish (Cyprinus carpio) were exposed to different concentrations of tebuconazol fungicide, by rice field (31.95 μg/L) and laboratory (33.47 and 36.23 μg/L) conditional testing, during a 7 day period. Parameters such thiobarbituric acid-reactive substance levels (TBARS), protein carbonyl, catalase, glutathione S-transferase and acetylcholinesterase activities were studied, using the liver, brain and white muscle of the fish. The field experiment showed that the TBARS levels were increased in all the analyzed tissues. Similarly, the protein carbonyl of the liver and the brain AChE activity increased after 7 days. The laboratory experiment demonstrated that the TBARS levels in the liver were increased in both of the concentration tests. TBARS levels in the muscle increased only by the lowest test concentration. On the other hand, the protein carbonyl was increased only by the highest concentration. The results indicate that the tebuconazol exposure from the field and laboratory conditions directly affected the health of the fish, showing the occurrence of oxidative stress.

  17. Genetic resources offer efficient tools for rice functional genomics research.

    Science.gov (United States)

    Lo, Shuen-Fang; Fan, Ming-Jen; Hsing, Yue-Ie; Chen, Liang-Jwu; Chen, Shu; Wen, Ien-Chie; Liu, Yi-Lun; Chen, Ku-Ting; Jiang, Mirng-Jier; Lin, Ming-Kuang; Rao, Meng-Yen; Yu, Lin-Chih; Ho, Tuan-Hua David; Yu, Su-May

    2016-05-01

    Rice is an important crop and major model plant for monocot functional genomics studies. With the establishment of various genetic resources for rice genomics, the next challenge is to systematically assign functions to predicted genes in the rice genome. Compared with the robustness of genome sequencing and bioinformatics techniques, progress in understanding the function of rice genes has lagged, hampering the utilization of rice genes for cereal crop improvement. The use of transfer DNA (T-DNA) insertional mutagenesis offers the advantage of uniform distribution throughout the rice genome, but preferentially in gene-rich regions, resulting in direct gene knockout or activation of genes within 20-30 kb up- and downstream of the T-DNA insertion site and high gene tagging efficiency. Here, we summarize the recent progress in functional genomics using the T-DNA-tagged rice mutant population. We also discuss important features of T-DNA activation- and knockout-tagging and promoter-trapping of the rice genome in relation to mutant and candidate gene characterizations and how to more efficiently utilize rice mutant populations and datasets for high-throughput functional genomics and phenomics studies by forward and reverse genetics approaches. These studies may facilitate the translation of rice functional genomics research to improvements of rice and other cereal crops.

  18. Role of peroxidases in the compensation of cytosolic ascorbate peroxidase knockdown in rice plants under abiotic stress.

    Science.gov (United States)

    Bonifacio, Aurenivia; Martins, Marcio O; Ribeiro, Carolina W; Fontenele, Adilton V; Carvalho, Fabricio E L; Margis-Pinheiro, Márcia; Silveira, Joaquim A G

    2011-10-01

    Current studies, particularly in Arabidopsis, have demonstrated that mutants deficient in cytosolic ascorbate peroxidases (APXs) are susceptible to the oxidative damage induced by abiotic stress. In contrast, we demonstrate here that rice mutants double silenced for cytosolic APXs (APx1/2s) up-regulated other peroxidases, making the mutants able to cope with abiotic stress, such as salt, heat, high light and methyl viologen, similar to non-transformed (NT) plants. The APx1/2s mutants exhibited an altered redox homeostasis, as indicated by increased levels of H₂O₂ and ascorbate and glutathione redox states. Both mutant and NT plants exhibited similar photosynthesis (CO₂) assimilation and photochemical efficiency) under both normal and stress conditions. Overall, the antioxidative compensatory mechanism displayed by the mutants was associated with increased expression of OsGpx genes, which resulted in higher glutathione peroxidase (GPX) activity in the cytosolic and chloroplastic fractions. The transcript levels of OsCatA and OsCatB and the activities of catalase (CAT) and guaiacol peroxidase (GPOD; type III peroxidases) were also up-regulated. None of the six studied isoforms of OsApx were up-regulated under normal growth conditions. Therefore, the deficiency in cytosolic APXs was effectively compensated for by up-regulation of other peroxidases. We propose that signalling mechanisms triggered in rice mutants could be distinct from those proposed for Arabidopsis.

  19. The never ripe mutant provides evidence that tumor-induced ethylene controls the morphogenesis of agrobacterium tumefaciens-induced crown galls on tomato stems

    Science.gov (United States)

    Aloni; Wolf; Feigenbaum; Avni; Klee

    1998-07-01

    We confirm the hypothesis that Agrobacterium tumefaciens-induced galls produce ethylene that controls vessel differentiation in the host stem of tomato (Lycopersicon esculentum Mill.). Using an ethylene-insensitive mutant, Never ripe (Nr), and its isogenic wild-type parent we show that infection by A. tumefaciens results in high rates of ethylene evolution from the developing crown galls. Ethylene evolution from isolated internodes carrying galls was up to 50-fold greater than from isolated internodes of control plants when measured 21 and 28 d after infection. Tumor-induced ethylene substantially decreased vessel diameter in the host tissues beside the tumor in wild-type stems but had a very limited effect in the Nr stems. Ethylene promoted the typical unorganized callus shape of the gall, which maximized the tumor surface in wild-type stems, whereas the galls on the Nr stems had a smooth surface. The combination of decreased vessel diameter in the host and increased tumor surface ensured water-supply priority to the growing gall over the host shoot. These results indicate that in addition to the well-defined roles of auxin and cytokinin, there is a critical role for ethylene in determining crown-gall morphogenesis.

  20. 水稻淡绿叶突变体 HM133的遗传分析与基因定位%Genetic Analysis and Gene Mapping of a Pale Green Leaf Mutant HM133 in Rice

    Institute of Scientific and Technical Information of China (English)

    施勇烽; 贺彦; 郭丹; 吕向光; 黄奇娜; 吴建利

    2016-01-01

    The pale green leaf mutant HM133 was identified from an EMS-induced IR64 mutant bank.The contents of photosynthetic pigments including chlorophyll and carotenoid of HM133 were reduced significantly at 6 weeks and 1 5 weeks after sowing when compared with IR64.The net photosynthetic rate of HM133 was considerably lower than that of the wild-type IR64 at heading stage while the stomatal conductance was apparently increased.The agronomic traits including plant height,number of filled grain per panicle and seed-setting rate decreased significantly in the mutant compared with the wild-type.In addition,the mutant exhibited a less number of grana,irregular arrangement of thylakoid layer in the chloroplast at the tillering stage.Genetic and mapping analysis showed that the pale green phenotype was controlled by a single recessive gene located in the long arm of chromosome 3 between SSR markers RM143 and RM3684.The interval contains an ORF OsChlD encoding magnesium-chelatase D subunit.Sequence analysis revealed that the mutant allele carried a nucleotide substitution from G to A in the tenth exon of OsChlD , which led to the substitution of glutamic acid for arginine acid.Therefore,it is deduced that OsChlD is the candidate gene controlling the pale green leaf phenotype of HM133 .%EMS 诱导籼稻品种 IR64获得淡绿叶突变体 HM133.与野生型 IR64相比,HM133播种后的第6周和第15周的光合色素含量以及抽穗期的净光合速率显著降低,气孔导度则明显上升;此外,突变体株高、每穗实粒数和结实率等农艺性状也较野生型显著下降.叶绿体超微结构分析表明,分蘖期 HM133类囊体基粒片层形状不规则,堆叠凌乱、排列疏松.遗传分析表明 HM133淡绿叶性状受单隐性核基因控制.通过分子标记将该基因定位于第3染色体长臂 RM143和 RM3684之间.该区间内包含编码镁螯合酶 D 亚基的基因 OsCHLD .序列分析表明 HM133中该基因第10外显子上有一个从 G

  1. Transcriptome and proteomic analyses reveal multiple differences associated with chloroplast development in the spaceflight-induced wheat albino mutant mta.

    Science.gov (United States)

    Shi, Kui; Gu, Jiayu; Guo, Huijun; Zhao, Linshu; Xie, Yongdun; Xiong, Hongchun; Li, Junhui; Zhao, Shirong; Song, Xiyun; Liu, Luxiang

    2017-01-01

    Chloroplast development is an integral part of plant survival and growth, and occurs in parallel with chlorophyll biosynthesis. However, little is known about the mechanisms underlying chloroplast development in hexaploid wheat. Here, we obtained a spaceflight-induced wheat albino mutant mta. Chloroplast ultra-structural observation showed that chloroplasts of mta exhibit abnormal morphology and distribution compared to wild type. Photosynthetic pigments content was also significantly decreased in mta. Transcriptome and chloroplast proteome profiling of mta and wild type were done to identify differentially expressed genes (DEGs) and proteins (DEPs), respectively. In total 4,588 DEGs including 1,980 up- and 2,608 down-regulated, and 48 chloroplast DEPs including 15 up- and 33 down-regulated were identified in mta. Classification of DEGs revealed that most were involved in chloroplast development, chlorophyll biosynthesis, or photosynthesis. Besides, transcription factors such as PIF3, GLK and MYB which might participate in those pathways were also identified. The correlation analysis between DEGs and DEPs revealed that the transcript-to-protein in abundance was functioned into photosynthesis and chloroplast relevant groups. Real time qPCR analysis validated that the expression level of genes encoding photosynthetic proteins was significantly decreased in mta. Together, our results suggest that the molecular mechanism for albino leaf color formation in mta is a thoroughly regulated and complicated process. The combined analysis of transcriptome and proteome afford comprehensive information for further research on chloroplast development mechanism in wheat. And spaceflight provides a potential means for mutagenesis in crop breeding.

  2. Unraveling rhizobacteria- and abscisic acid-induced pathogen resistance in rice (Oryza sativa L.)

    OpenAIRE

    De Vleesschauwer, David

    2008-01-01

    Induced disease resistance is the phenomenon by which plants exhibit a heightened level of resistance against pathogen infection after appropriate stimulation. In contrast to the relative wealth of information on inducible resistance responses in dicotyledoneous plants, our current understanding of the molecular machinery governing induced resistance in monocotyledoneous crops is still in its infancy. In light of aforementioned knowledge gap, this dissertation aimed to expand our knowledge on...

  3. 水稻株型突变体rad-1和rad-2的鉴定与功能基因克隆%Identification and Map-based Cloning ofrad-1 andrad-2, Two Rice Architec-ture Determinant Mutants

    Institute of Scientific and Technical Information of China (English)

    牛静; 陈赛华; 赵婕妤; 曾召琼; 蔡茂红; 周亮; 刘喜; 江玲; 万建民

    2015-01-01

    株型是决定水稻等作物产量的核心因素之一,是品种选育的重要指标。本研究从粳稻品种Asominori的辐射诱变中分离出2个稳定的株型突变体, rad-1和rad-2,它们均表现出苗期弯曲生长、成熟期株高矮化、粒长变短、千粒重降低和产量下降等特征。等位性测验结合连锁分析证实rad-1和rad-2等位,且位于水稻第7染色体上约230 kb的范围内。对定位区段的序列分析后确定OsFH5为候选基因,该基因呈现组成型表达,编码水稻II型成蛋白。突变体rad-1在OsFH5基因第2个外显子上缺失8个碱基,导致移码;而rad-2则在第14内含子上发生单碱基的变异,发生异常剪切。两类突变最终均导致OsFH5基因翻译提前终止,产生截短的蛋白。相比野生型, rad-1和rad-2在幼苗中OsFH5基因的表达下调。细胞学研究表明, OsFH5基因的功能缺失会导致幼苗叶鞘细胞大小不均,呈现不规则生长,在成熟的颖壳中细胞显著变短。对生长素响应的 ARF因子进行表达量检测发现, rad-2中一系列 ARF成员表达均显著下调,推测OsFH5极有可能影响了植株对生长素的响应。%Plant architecture is a key element of crop yield and also taken as a criterion in breeding. In this study, two rice archi-tecture determinant mutants,rad-1 andrad-2 were selected from Gamma-radiation induced mutants of rice cultivar Asominori. Each of them shows winding seedlings, dwarf plants and thereby decreased yield. It has been estimated thatrad-1 andrad-2 are allelic to each other by allelism test and linkage analysis. After map-based cloning, the responsible gene for the mutations was restricted within a 230 kb region in chromosome 7. Sequence analysis revealed thatOsFH5 gene, encoding a forming-like protein altered both inrad-1 andrad-2. An 8-bp deletion in exon 2 inrad-1 and a single nucleotide change in intron 14 inrad-2 made frame-shift reading and produced truncated proteins.OsFH5 gene

  4. Ghd2, a CONSTANS-like gene, confers drought sensitivity through regulation of senescence in rice

    Science.gov (United States)

    Liu, Juhong; Shen, Jianqiang; Xu, Yan; Li, Xianghua; Xiao, Jing