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Sample records for repeats issr markers

  1. Application of inter simple sequence repeat (ISSR) markers to plant genetics.

    Science.gov (United States)

    Godwin, I D; Aitken, E A; Smith, L W

    1997-08-01

    Microsatellites or simple sequence repeats (SSRs) are ubiquitous in eukaryotic genomes. Single-locus SSR markers have been developed for a number of species, although there is a major bottleneck in developing SSR markers whereby flanking sequences must be known to design 5'-anchors for polymerase chain reaction (PCR) primers. Inter SSR (ISSR) fingerprinting was developed such that no sequence knowledge was required. Primers based on a repeat sequence, such as (CA)n, can be made with a degenerate 3'-anchor, such as (CA)8RG or (AGC)6TY. The resultant PCR reaction amplifies the sequence between two SSRs, yielding a multilocus marker system useful for fingerprinting, diversity analysis and genome mapping. PCR products are radiolabelled with 32P or 33P via end-labelling or PCR incorporation, and separated on a polyacrylamide sequencing gel prior to autoradiographic visualisation. A typical reaction yields 20-100 bands per lane depending on the species and primer. We have used ISSR fingerprinting in a number of plant species, and report here some results on two important tropical species, sorghum and banana. Previous investigators have demonstrated that ISSR analysis usually detects a higher level of polymorphism than that detected with restriction fragment length polymorphism (RFLP) or random amplified polymorphic DNA (RAPD) analyses. Our data indicate that this is not a result of greater polymorphism genetically, but rather technical reasons related to the detection methodology used for ISSR analysis.

  2. Genetic diversity analysis of Lepidium sativum (Chandrasur) using inter simple sequence repeat (ISSR) markers

    Institute of Scientific and Technical Information of China (English)

    Amandeep Kaur; Rakesh Kumar; Suman Rani; Anita Grewal

    2015-01-01

    Lepidium sativum (commonly known as garden cress) belongs to the family Brassicaceae. It is a fast-growing erect, annual herbaceous plant. Its seeds possess significant fracture healing, anti-asthmatic, anti-diabetic, hypoglycemic, nephrocurative and nephroprotective activ-ities. In the present study, we assessed the genetic diversity of various genotypes of L. sativum using inter-simple sequence repeat (ISSR) markers. Out of 41 ISSR primers screened, 32 primers showed significant, clear and repro-ducible bands. A total of 510 amplified bands were obtained using 32 ISSR primers, out of which 422 bands were poly-morphic and 88 bands were monomorphic. The percentage of polymorphism was found to be 82. A total of 35 unique alleles ranging insize from 200 to 2,900 bp were observed. Cluster analysis based on unweighted pair-group method, arithmetic mean divided the 18 genotypes into two main clusters, with the first having only HCS-08 genotype of L. sativum and other having all of the other 17 genotypes. The Jaccard similarity coefficient revealed a broad range 32–72%genetic relatedness among the 18 genotypes.

  3. Molecular diversity and relationships among Cymbidium goeringii cultivars based on inter-simple sequence repeat (ISSR) markers.

    Science.gov (United States)

    Wang, Hui-Zhong; Wu, Zhen-Xing; Lu, Jiang-Jie; Shi, Nong-Nong; Zhao, Yan; Zhang, Zhi-Tao; Liu, Jun-Jun

    2009-07-01

    Spring orchid (Cymbidium goeringii) is a popular flowering plant species. There have been few molecular studies of the genetic diversity and conservation genetics on this species. An assessment of the level of genetic diversity in cultivated spring orchid would facilitate development of the future germplasm conservation for cultivar improvement. In the present study, DNA markers of intersimple sequence repeats (ISSR) were identified and the ISSR fingerprinting technique was used to evaluate genetic diversity in C. goeringii cultivars. Twenty-five ISSR primers were selected to produce a total of 224 ISSR loci for evaluation of the genetic diversity. A wide genetic variation was found in the 50 tested cultivars with Nei's gene diversity (H = 0.2241) and 93.75% of polymorphic loci. Fifty cultivars were unequivocally distinguished based on ISSR fingerprinting. Cultivar-specific ISSR markers were identified in seven of 50 tested cultivars. Unweighted pair-group mean analysis (UPGMA) and principal coordinates analysis (PCA) grouped them into two clusters: one composed the cultivars mainly from Japan, and the other contained three major subclusters mainly from China. Two Chinese subclusters were generally consistent with horticultural classification, and the third Chinese subcluster contained cultivars from various horticultural groups. Our results suggest that the ISSR technique provides a powerful tool for cultivar identification and establishment of genetic relationships of cultivars in C. goeringii.

  4. Analysis of genetic relationship in mutant silkworm strains of Bombyx mori using inter simple sequence repeat (ISSR) markers

    Institute of Scientific and Technical Information of China (English)

    Dhanikachalam Velu; Kangayam M. Ponnuvel; Murugiah Muthulakshmi; Randhir K. Sinha; Syed M.H. Qadri

    2008-01-01

    Amplified inter simple sequence repeats (ISSR) markers were used to determine genetic relationships among mutant silkworm strains of Bombyx mori. Fifteen ISSR primers containing simple sequence repeat (SSR) motifs were used in this study. A total of 113 markers were produced among 20 mutant swains, of which 73.45% were found to be polymorphic. In selected mutant genetic stocks, the average number of observed allele was (1.7080±0.4567), effective alleles (1.5194±0.3950) and genetic diversity (Ht) (0.2901±0.0415). The dendrogram produced using the unweighted pair group method with arithmetic means (UPGMA) and cluster analysis made using Nei's genetic distance resulted in the formation of one major group containing 6 groups separated 20 mutant silkworm strains. Therefore, ISSR amplification is a valuable method for determining the genetic variability among mutant silkworm swains. This efficient molecular marker would be useful for characterizing a considerable number of silkworm swains maintained at the germplasm center.

  5. Inter-Simple Sequence Repeat (ISSR Markers to Study Genetic Diversity Among Cotton Cultivars in Associated with Salt Tolerance

    Directory of Open Access Journals (Sweden)

    Ali Akbar ABDI

    2012-11-01

    Full Text Available Developing salt-tolerant crops is very important as a significant proportion of cultivated land is salt-affected. Screening and selection of salt tolerant genotypes of cotton using DNA molecular markers not only introduce tolerant cultivars useful for hybridization and breeding programs but also detect DNA regions involved in mechanism of salinity tolerance. To study this, 28 cotton cultivars, including 8 Iranian cotton varieties were grown in pots under greenhouse condition and three salt treatments were imposed with salt solutions (0, 70 and 140 mM NaCl. Eight agronomic traits including root length, root fresh weight, root dry weight, chlorophyll and fluorescence index, K+ and Na+ contents in shoot (above ground biomass, and K+/Na+ ratio were measured. Cluster analysis of cultivars based on measured agronomic traits, showed �Cindose� and �Ciacra� as the most tolerant cultivars, and �B-557� and �43347� as the most sensitive cultivars of salt damage. A total of 65 polymorphic DNA fragments were generated at 14 inter-simple sequence repeat (ISSR loci. Plants of 28 cultivars of cotton grouped into three clusters based on ISSR markers. Regression analysis of markers in relation with traits data showed that 23, 33 and 30 markers associated with the measured traits in three salt treatments respectively. These markers might help breeders in any marker assisted selection program in order to improving cotton cultivars against salt stress.

  6. Diversity and genetic stability in banana genotypes in a breeding program using inter simple sequence repeats (ISSR) markers.

    Science.gov (United States)

    Silva, A V C; Nascimento, A L S; Vitória, M F; Rabbani, A R C; Soares, A N R; Lédo, A S

    2017-02-23

    Banana (Musa spp) is a fruit species frequently cultivated and consumed worldwide. Molecular markers are important for estimating genetic diversity in germplasm and between genotypes in breeding programs. The objective of this study was to analyze the genetic diversity of 21 banana genotypes (FHIA 23, PA42-44, Maçã, Pacovan Ken, Bucaneiro, YB42-47, Grand Naine, Tropical, FHIA 18, PA94-01, YB42-17, Enxerto, Japira, Pacovã, Prata-Anã, Maravilha, PV79-34, Caipira, Princesa, Garantida, and Thap Maeo), by using inter-simple sequence repeat (ISSR) markers. Material was generated from the banana breeding program of Embrapa Cassava & Fruits and evaluated at Embrapa Coastal Tablelands. The 12 primers used in this study generated 97.5% polymorphism. Four clusters were identified among the different genotypes studied, and the sum of the first two principal components was 48.91%. From the Unweighted Pair Group Method using Arithmetic averages (UPGMA) dendrogram, it was possible to identify two main clusters and subclusters. Two genotypes (Garantida and Thap Maeo) remained isolated from the others, both in the UPGMA clustering and in the principal cordinate analysis (PCoA). Using ISSR markers, we could analyze the genetic diversity of the studied material and state that these markers were efficient at detecting sufficient polymorphism to estimate the genetic variability in banana genotypes.

  7. Application of ISSR marker in pharmacognosy: Current update

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    Jayvant Kurane

    2009-01-01

    Full Text Available ISSR (Inter Simple Sequence Repeat is one of the popular techniques of DNA fingerprinting because of several reasons. In many fields, ISSR markers have proved their utility. There are many applications of ISSR in various aspects of medicinal plants. ISSR based markers have utility in the fields like genetics, taxonomy, physiology, embryology etc. and recently the ISSR based markers have found wide applicability in pharmacognostic characterization of medicinal plants. As use of herbal medicines is increasing, there is urgent need of newer technologies and its proper application. In recent years, pharmacognosy has witnessed advent of such new technologies. This review provides detail list of plants, which are studied by ISSR marker and discuss some of the important application in medicinal plant research.

  8. [ISSR markers and their applications in plant genetics].

    Science.gov (United States)

    Wang, Jian-bo

    2002-09-01

    Recently, inter-simple sequence repeat (ISSR) markers have emerged as an alternative system with reliability and advantages of microsatellites (SSR). The technique involves amplification of genomic segments flanked by inversely oriented and closely spaced microsatellite sequences by a single primer or a pair of primers based on SSRs anchored 5' or 3' with 1-4 purine or pyramidine residues. The sequences of repeats and anchor nucleates are arbitrarily selected. Coupled with the separation of amplification products on a polyacrylamide or agarose gels,ISSR amplification can reveal a much larger number of fragments per primer than RAPD. It is concluded that ISSR technique provides a quick, reliable and highly informative system for DNA fingerprinting.ISSR markers are inherited in Mendelin mode and segregated as dominant markers. This technique has been widely used in the studies of cultivar identification, genetic mapping, gene tagging,genetic diversity, evolution and molecular ecology.

  9. ISSR markers based on GA and AG repeats reveal genetic relationship among rice varieties tolerant to drought,flood,or salinity

    Institute of Scientific and Technical Information of China (English)

    Ch Surendhar REDDY; A.Prasad BABU; B.P.Mallikarjuna SWAMY; K.KALADHAR; N.SARLA

    2009-01-01

    Drought,flood,salinity,or a combination of these limits rice production.Several rice varieties are well known for their tolerance to specific abiotic stresses.We determined genetic relationship among 12 rice varieties including 9 tolerant to drought,flood,or salinity using inter-simple sequence repeat (ISSR) markers.Based on all markers,the nine tolerant varieties formed one cluster distinct from the cluster of three control varieties.The salt-tolerant varieties were closest to two flood-tolerant varieties,and together they were distinct from the drought-tolerant varieties.(GA)8 YG was the most informative primer,showing the highest polymorphic information content (PIC) and resolving power (Rp).The drought-,flood-,and salt-tolerant varieties grouped in three distinct clusters within the group of tolerant varieties,when (GA)8 YG was used.Sabita was the only exception.The two aus varieties,Nagina22 and FR13A,were separated and grouped with the drought-and flood-tolerant varieties,respectively,but they were together in dendrograms based on other primers.The results show that ISSR markers associated with (GA)8 YG delineated the three groups of stress-tolerant varieties from each other and can be used to identify genes/new alleles associated with the three abiotic stresses in rice germplasm.

  10. Genetic variability and geographic differentiation in Thymus daenensis subsp. daenensis, an endangered medicinal plant, as revealed by inter simple sequence repeat (ISSR) markers.

    Science.gov (United States)

    Rahimmalek, Mehdi; Bahreininejad, Babak; Khorrami, Mojtaba; Tabatabaei, Badraldin Ebrahim Sayed

    2009-12-01

    Thymus daenensis is an aromatic medicinal plant endemic to Iran. We used inter simple sequence repeat (ISSR) markers to detect genetic polymorphism in this herb using 17 T. daenensis accessions collected from different geographic regions in Iran. The 15 primers chosen for analysis revealed 256 bands, of which 228 (88.9%) were polymorphic. Jaccard's similarity indices based on ISSR profiles were subjected to UPGMA cluster analysis. The generated dendrogram revealed two major groups. The Tc group included the accessions collected from the center of the Zagros Mountains, and the Te group was collected from the extremes of the Zagros range. A principal coordinate analysis confirmed the results of clustering. The results showed that the divergence of accessions based on the Zagros Mountains is more logical in comparison with classification on the basis of provincial borders. Gene diversity and expected heterozygosity were greater in the Tc group than in the Te group, suggesting that the germplasm collected from the center of the Zagros Mountains is more variable.

  11. Genetic variation of the genus Kengyilia by ISSR markers

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    We investigated the genetic variation within 32 accessions distributed to 14 species and one variety by using ISSR (inter-simple sequence repeat) markers.The results showed that genetic variation was relatively higher among the accessions.A total of 593 bands were amplified by 12 ISSR primers,of which 535 bands (90.2%) were polymorphic.Eleven to 80 polymorphie bands were amplified from each prime,with an average of 44.6 bands.The interspecies GS (genetic similarity)value ranged from 0.430 to 0.866,and the average was 0.620.Cluster analysis showed that all accessions could be classified into 4 groups by ISSR markers.The different accessions in a species were clustered together,but they had genetic variation in molecular levels.There was obvious interspecies genetic variation.Species with similar morphological characteristics and from the same areas or neighboring geographical regions were clustered together and had close relationships.ISSR markers are useful in analyzing interspecies variation in Kengyilia.

  12. Genetic Variation in Five Mediterranean Populations of Juniperus phoenicea as Revealed by Inter-Simple Sequence Repeat (ISSR) Markers

    Science.gov (United States)

    MELONI, MARILENA; PERINI, DAVIDE; FILIGHEDDU, ROSSELLA; BINELLI, GIORGIO

    2006-01-01

    • Background and Aims The assessment of the genetic variability and the identification of isolated populations within a given species represent important information to plan conservation strategies on a genetic basis. In this work, the genetic variability in five natural populations of Juniperus phoenicea, three from Sardinia, one from Cyprus and the last one in the Maritime Alps was analysed by means of ISSRs, on the hypothesis that the latter could have been a refugial one during the last glaciation. • Methods ISSRs were chosen because of their ability to detect variation without any prior sequence information. The use of three primers yielded 45 reproducible, polymorphic bands, which were utilized to estimate the basic parameters of genetic variability and diversity. • Key Results All of the populations analysed harboured an adequate amount of genetic variability, with HS = 0·1299. The proportion of genetic diversity between populations has been estimated by GST = 0·12. The three Sardinian populations are separated, as tested by AMOVA, from the Cyprus and the continental ones. • Conclusions The results indicate that geographical isolation has represented a major barrier to gene flow in Juniperus phoenicea. This work represents a first step towards a full genetic characterization of a conifer from the Mediterranean, a world biodiversity hotspot confronted with climate change, and thus contributes towards the planning of genetics-informed conservation strategies. PMID:16311272

  13. Individual and population variation in invertebrates revealed by Inter-simple Sequence Repeats (ISSRs

    Directory of Open Access Journals (Sweden)

    Patrick Abbot

    2001-08-01

    Full Text Available PCR-based molecular markers are well suited for questions requiring large scale surveys of plant and animal populations. Inter-simple Sequence Repeats or ISSRs are analyzed by a recently developed technique based on the amplification of the regions between inverse-oriented microsatellite loci with oligonucleotides anchored in microsatellites themselves. ISSRs have shown much promise for the study of the population biology of plants, but have not yet been explored for similar studies of animals. The value of ISSRs is demonstrated for the study of animal species with low levels of within-population variation. Sets of primers are identified which reveal variation in two aphid species, Acyrthosiphon pisum and Pemphigus obesinymphae, in the yellow fever mosquito Aedes aegypti, and in a rotifer in the genus Philodina.

  14. Identification of necrophagous fly species from 12 different cities in China using ISSR and SCAR markers

    Institute of Scientific and Technical Information of China (English)

    Xueli Zheng; Jialin Hu; Santhosh Puthiya Kunnon; Chen Xiaoguang

    2010-01-01

    Objective:To identify necrophagous fly speies from different regions in China using inter simple sequenc repeat (ISSR) and sequence-characterized amplified region (SCAR) melocular markers and to analyze their gene difference and genetic relationship. Methods:Five carrion fly species were collected from 12 cities and regions in China, including Musca domestica (M. domestica), Lucilia sericata (L. sericata), Chrysomya megacephala (C. megacephala), Helicophagella melanura (H. melanura), Boethcherisca peregrina, and they were studied using ISSR and SCAR markers. Results:Eight ISSR primers were used for amplification of 121 samples. 679 clear and stable bands were identified, of which 516 bands were polymorphic. Several species-specific ISSR fragment were cloned and sequenced as an initial effort to derive the SCAR markers. Using M. domestica SCAR specific primers, SCAR-PCR amplification was performed for 8 M. domestca population sample DNA from different regions in China as well as L. sericata, C. megacephala, H. melanura and Lucillia cupirina. The result showed only M. domestica produced specificalty 600 bp fragment, but L. sericata, C. megacephala, H. melanura and Lucillia cupirina did not produce the same specific fragment. Clustering analysis showed clustering of most flies of M. domestica, C. megacephala and L. sericata. M. domestica samples from different regions in China yielded different banding patterns. Conclusions:Application of ISSR-PCR and SCAR markers to identify necrophagous fly species from 12 cities and regions in China is first reported. ISSR-PCR and SCAR markers provide a quick reliable molecular marker technique for the identification of different species of necrophagous fly.

  15. Genetic variability among 18 cultivars of cooking bananas and plantains by RAPD and ISSR markers

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    YUYU SURYASARI POERBA

    2010-07-01

    Full Text Available Poerba YS, Ahmad F (2010 Genetic variability among 18 cultivars of cooking bananas and plantains by RAPD and ISSR markers. Biodiversitas 11: 118-123. This study was done to assess the molecular diversity of 36 accessions (18 cultivars of the plantain and cooking bananas (Musa acuminata x M. balbisiana, AAB, ABB subgroups based on Random amplified polymorphic DNA (RAPD and and Inter Simple Sequence Repeats (ISSR markers and to determine genetic relationships in the bananas. RAPD and ISSR fingerprinting of these banana varieties was carried out by five primers of RAPDs and two primers of ISSRs. RAPD primers produced 63 amplified fragments varying from 250 to 2500 bp in size. 96.82% of the amplification bands were polymorphic. ISSR primers produced 26 amplified fragments varying from 350 bp to 2000 bp in size. The results showed that 92.86% of the amplification bands were polymorphic. The range of genetic distance of 18 cultivars was from 0.06-0.67.

  16. Evaluation of genetic diversity of Clinacanthus nutans (Acanthaceaea) using RAPD, ISSR and RAMP markers.

    Science.gov (United States)

    Ismail, Noor Zafirah; Arsad, Hasni; Samian, Mohammed Razip; Ab Majid, Abdul Hafiz; Hamdan, Mohammad Razak

    2016-10-01

    Three polymerase chain reaction (PCR) techniques were compared to analyse the genetic diversity of Clinacanthus nutans eight populations in the northern region of Peninsular Malaysia. The PCR techniques were random amplified polymorphic deoxyribonucleic acids (RAPD), inter-simple sequence repeats (ISSR) and random amplified microsatellite polymorphisms (RAMP). Leaf genomic DNA was PCR amplified using 17 RAPD, 8 ISSR and 136 RAMP primers . However, only 10 RAPD primers, 5 ISSR primers and 37 RAMP primers produced reproducible bands. The results were evaluated for polymorphic information content (PIC), marker index (MI) and resolving power (RP). The RAMP marker was the most useful marker compared to RAPD and ISSR markers because it showed the highest average value of PIC (0.25), MI (11.36) and RP (2.86). The genetic diversity showed a high percentage of polymorphism at the species level compared to the population level. Furthermore, analysis of molecular variance revealed that the genetic diversity was higher within populations, as compared to among populations of C. nutans. From the results, the RAMP technique was recommended for the analysis of genetic diversity of C. nutans.

  17. Construction of intersubspecific molecular genetic map of lentil based on ISSR, RAPD and SSR markers

    Indian Academy of Sciences (India)

    Mamta Gupta; Bhawna Verma; Naresh Kumar; Rakesh K. Chahota; Rajeev Rathour; Shyam K. Sharma; Sabhyata Bhatia; Tilak R. Sharma

    2012-12-01

    Lentil (Lens culinaris ssp. culinaris), is a self-pollinating diploid ($2n = 2x = 14$), cool-season legume crop and is consumed worldwide as a rich source of protein (∼24.0%), largely in vegetarian diets. Here we report development of a genetic linkage map of Lens using 114 F2 plants derived from the intersubspecific cross between L 830 and ILWL 77. RAPD (random amplified polymorphic DNA) primers revealed more polymorphism than ISSR (intersimple sequence repeat) and SSR (simple sequence repeat) markers. The highest proportion (30.72%) of segregation distortion was observed in RAPD markers. Of the 235 markers (34 SSR, 9 ISSR and 192 RAPD) used in the mapping study, 199 (28 SSRs, 9 ISSRs and 162 RAPDs) were mapped into 11 linkage groups (LGs), varying between 17.3 and 433.8 cM and covering 3843.4 cM, with an average marker spacing of 19.3 cM. Linkage analysis revealed nine major groups with 15 or more markers each and two small LGs with two markers each, and 36 unlinked markers. The study reported assigning of 11 new SSRs on the linkage map. Of the 66 markers with aberrant segregation, 14 were unlinked and the remaining 52 were mapped. ISSR and RAPD markers were found to be useful in map construction and saturation. The current map represents maximum coverage of lentil genome and could be used for identification of QTL regions linked to agronomic traits, and for marker-assisted selection in lentil.

  18. Inter-simple sequence repeat (ISSR) loci mapping in the genome of perennial ryegrass

    DEFF Research Database (Denmark)

    Pivorienė, O; Pašakinskienė, I; Brazauskas, G;

    2008-01-01

    The aim of this study was to identify and characterize new ISSR markers and their loci in the genome of perennial ryegrass. A subsample of the VrnA F2 mapping family of perennial ryegrass comprising 92 individuals was used to develop a linkage map including inter-simple sequence repeat markers...... demonstrated a 70% similarity to the Hordeum vulgare germin gene GerA. Inter-SSR mapping will provide useful information for gene targeting, quantitative trait loci mapping and marker-assisted selection in perennial ryegrass....

  19. Genetic variation assessment of acid lime accessions collected from south of Iran using SSR and ISSR molecular markers.

    Science.gov (United States)

    Sharafi, Ata Allah; Abkenar, Asad Asadi; Sharafi, Ali; Masaeli, Mohammad

    2016-01-01

    Iran has a long history of acid lime cultivation and propagation. In this study, genetic variation in 28 acid lime accessions from five regions of south of Iran, and their relatedness with other 19 citrus cultivars were analyzed using Simple Sequence Repeat (SSR) and Inter-Simple Sequence Repeat (ISSR) molecular markers. Nine primers for SSR and nine ISSR primers were used for allele scoring. In total, 49 SSR and 131 ISSR polymorphic alleles were detected. Cluster analysis of SSR and ISSR data showed that most of the acid lime accessions (19 genotypes) have hybrid origin and genetically distance with nucellar of Mexican lime (9 genotypes). As nucellar of Mexican lime are susceptible to phytoplasma, these acid lime genotypes can be used to evaluate their tolerance against biotic constricts like lime "witches' broom disease".

  20. Molecular identification of Aquilaria spp. by using inter-simple sequence repeat (ISSR)

    Science.gov (United States)

    Azhari, Hanif; Mohamad, Azhar; Othman, Roohaida

    2015-09-01

    Aquilaria species are very important economic plant for production of resin locally known as gaharu in Malaysia. There are five species that can be found in Malaysia and the most important Aquilaria species for gaharu production is A. malaccensis. Molecular markers for Aquilaria species are still insufficient and require more efficient, robust and reproducible molecular marker. Inter-simple sequence repeat (ISSR) markers are highly polymorphic and have high reproducibility which will be useful in areas of genetic diversity, phylogenetic studies, gene tagging, genome mapping and evolutionary biology in a wide range of crop species. Five selected ISSR primers were used to identify four Aquilaria species commonly found in Malaysia namely A. malaccensis, A. sub-integra, A. crassna and A. hirta. All the primers showed sufficient polymorphism to distinguish between the four species. Hence, the markers derived from ISSR can be used for molecular identification of Aquilaria spp. in ensuring homogenous species for plantation which may improve the quality of resin derived from known and certified materials.

  1. Identification of Syringa oblata by Inter-Simple Sequence Repeat Markers%白花与紫花丁香ISSR-PCR鉴别研究

    Institute of Scientific and Technical Information of China (English)

    思彬彬; 赵海燕; 刘海涛

    2012-01-01

    [ Objective] To identify Syringa oblata by inter-simple sequence repeat markers. [ Method] Primers suitable for routine analysis were screened from 100 inter-simple sequence repeat primers, then, they were used in PCR and separated of 2 samples of Syringa oblata. [ Results ] Three of the one-hundred primers amplified polymorphic bands and suitable for the identification of Syringa oblata. [Conclusion] Inter-simple sequence repeat markers provide a quick, reliable molecular marker for identification of Syringa oblata.%[目的]探索用ISSR分子标记方法在核酸分子水平上鉴别白花与紫花丁香.[方法]从100条ISSR引物中筛选合适的引物对白花和紫花丁香2个样品进行PCR扩增及电泳分析,寻找特征位点.[结果]有3条ISSR引物扩增出较为明显的多态性特征条带,可单独应用于白花和紫花丁香的鉴别.[结论] ISSR作为一种简便、可靠的分子标记方法,可用于不同花色丁香的鉴别.

  2. GENETIC RELATIONSHIPS OF FIVE PERITRICHOUS CILIATES INFERRED FROM INTER SIMPLE SEQUENCE REPEAT (ISSR) MOLECULAR MARKERS%五种缘毛类纤毛虫遗传关系的ISSR研究

    Institute of Scientific and Technical Information of China (English)

    张文静; 余育和; 沈韫芬

    2005-01-01

    In this study, ISSR (inter simple sequence repeat) molecular markers were used to clarify genetic relationships of five peritrichous ciliates: Carchesium polypinum, Epistylis chrysemydis, E. plicatilis, E. urceolata and Vorticella campanula.From 34 primers, 13 polymorphic primers were selected and tested. The genetic distance values going from 0.6667 to 1. 0000 among the five species show the ISSR-PCR method has a high resolution. In the phylogenetic tree constructed with the unweighted pair group method using arithmetic averages (UPGMA), V. campanula was separated from other species firstly; C.polypinum and E. chrysemydis were closely related; in the group of Epistylis species, E. plicatilis and E. urceolata were first clustered together and then grouped with E. chrysemydis. These results were compared with the phylogenetic tree from the complete small subunit ribosomal RNA (SSrRNA) gene sequences. We found that: 1) ISSR primers can give the informative profiles in peritrichous ciliates; 2) C. polypinum has a nearer relation with Epistylis than V. campanula. The colonial or solitary status might be a more important phylogenetic character within peritrichs than the characters of stalk and the myoneme; 3) E.urceolata shared closer relationship with E. plicatilis than E. chrysemydis. The hollow nature of the stalk is a useful phylogenetic and taxonomic character in the genus Epistylis. This study demonstrated that ISSR-PCR method was a new useful method to reveal the relationships among related or similar species in ciliates.%应用ISSR分子标记揭示了5种缘毛类纤毛虫(Carchesium polypinum,Epistylis chrysemydis,E.plicatilis,E.urceolata和Vorticella campanula)的遗传关系.从34个引物中筛选到13个多态性高的引物进行研究.得到的遗传距离(0.666 7~1.000 0)显示ISSR技术具有较高的分辨率.依据构建的UPGMA聚类树,V.campanula首先和其他种类分开;C.polypinum和E.chrysemydis聚在了一起;在3种Epistylis纤毛虫中,E

  3. Preliminary Comparative Analysis of Phenological Varieties of Quercus robur by ISSR-Markers

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    Vasiliy Chokheli

    2016-01-01

    Full Text Available Quercus robur L. is a valuable wood species having long ontogeny and promising to create long-living artificial plantings of recreational and ameliorative purposes in the steppes zone of Russia and other countries. In this work we have performed the genotyping of varieties of Quercus robur L. obtained from collection of Botanical Garden of Southern Federal University using intersimple sequence repeat (ISSR molecular markers. The most polymorphic ISSR-marker (GA 8YC was found in the collection. The polymorphic DNA markers identified in the present study can be used for the future breeding works to obtain valuable genotypes of Quercus genus. In addition we have performed DNA fingerprinting of the prospective sample of the variety Q. robur var. tardiflora Czern.

  4. The study of genetic diversity of Daemonorops draco (Palmae using ISSR markers

    Directory of Open Access Journals (Sweden)

    REVIS ASRA

    2014-10-01

    Full Text Available Asra R, Syamsuardi, Mansyurdin, Witono JR. 2014. The study of genetic diversity of Daemonorops draco (Palmae using ISSR markers. Biodiversitas 15: 109-114. The genetic diversity in five populations of Daemonorops draco(Willd. Blume (Jernang: in Bahasa Indonesia was analyzed using Inter Simple Sequence Repeat (ISSR markers. The screening results from using 15 ISSR primers showed that only 5 of ISSR primers had clear and reproducible bands. Based on the data from the matrix binary analyzed using POPGENE version 3.2, the highest genetic diversity was found in the Sepintun population at 0.0969 average heterozygosis (H and 0.146 average Shannon Index (I. The heterozygosis calculation of the total population (HT was 0.2571. The heterozygosis value within a population (HS=0.0704 was smaller than that between populations (DST=0.1867. Using the clustering analysis program Past version 32 on 43 individuals of D. draco, we found that there were three groups of D. draco. Group A consisted of 8 individuals in the Bengayoan population, group B consisted of 9 units in the Nunusan population and group C consisted of three populations; Tebo, Sepintun and Mandiangin consisted of 10, 8 and 8 individuals. The genetic similarity varied among all populations withthe values between 0.07-0.93.

  5. Population genetics of Sargassum horneri (Fucales, Phaeophyta) in China revealed by ISSR and SRAP markers

    Science.gov (United States)

    Yu, Shenhui; Chong, Zhuo; Zhao, Fengjuan; Yao, Jianting; Duan, Delin

    2013-05-01

    Sargassum horneri is a common brown macro-alga that is found in the inter-tidal ecosystems of China. To investigate the current status of seaweed resources and provide basic data for its sustainable development, ISSR (inter simple sequence repeat) and SRAP (sequence related amplified polymorphism) markers were used to analyze the population genetics among nine natural populations of S. horneri. The nine studied populations were distributed over 2 000 km from northeast to south China. The percentage of polymorphic loci P % (ISSR, 99.44%; SRAP, 100.00%), Nei's genetic diversity H (ISSR, 0.107-0.199; SRAP, 0.100-0.153), and Shannon's information index I (ISSR, 0.157-0.291; SRAP, 0.148-0.219) indicated a fair amount of genetic variability among the nine populations. Moreover, the high degree of gene differentiation G st (ISSR, 0.654; SRAP, 0.718) and low gene flow N m (ISSR, 0.265; SRAP, 0.196) implied that there was significant among-population differentiation, possibly as a result of habitat fragmentation. The matrices of genetic distances and fixation indices ( F st) among the populations correlated well with their geographical distribution (Mantel test R =0.541 5, 0.541 8; P =0.005 0, 0.002 0 and R =0.728 6, 0.641 2; P =0.001 0, 0.001 0, respectively); the Rongcheng population in the Shandong peninsula was the only exception. Overall, the genetic differentiation agreed with the geographic isolation. The fair amount of genetic diversity that was revealed in the S. horneri populations in China indicated that the seaweed resources had not been seriously affected by external factors.

  6. Population genetics of Sargassum horneri (Fucales,Phaeophyta) in China revealed by ISSR and SRAP markers

    Institute of Scientific and Technical Information of China (English)

    YU Shenhui; CHONG Zhuo; ZHAO Fengjuan; YAO Jianting; DUAN Delin

    2013-01-01

    Sargassum horneri is a common brown macro-alga that is found in the inter-tidal ecosystems of China.To investigate the current status of seaweed resources and provide basic data for its sustainable development,ISSR (inter simple sequence repeat) and SRAP (sequence related amplified polymorphism)markers were used to analyze the population genetics among nine natural populations of S.horneri.The nine studied populations were distributed over 2 000 km from northeast to south China.The percentage of polymorphic loci P% (ISSR,99.44%; SRAP,100.00%),Nei's genetic diversity H(ISSR,0.107-0.199; SRAP,0.100-0.153),and Shannon's information index I (ISSR,0.157-0.291; SRAP,0.148-0.219) indicated a fair amount of genetic variability among the nine populations.Moreover,the high degree of gene differentiation Gst (ISSR,0.654; SRAP,0.718) and low gene flow Nm (ISSR,0.265; SRAP,0.196) implied that there was significant among-population differentiation,possibly as a result of habitat fragmentation.The matrices of genetic distances and fixation indices (Fst) among the populations correlated well with their geographical distribution (Mantel test R=0.541 5,0.541 8; P=0.005 0,0.002 0 and R=0.728 6,0.641 2; P=0.001 0,0.001 0,respectively); the Rongcheng population in the Shandong peninsula was the only exception.Overall,the genetic differentiation agreed with the geographic isolation.The fair amount of genetic diversity that was revealed in the S.horneri populations in China indicated that the seaweed resources had not been seriously affected by external factors.

  7. Identification and Validation of a New Male Sex-Specific ISSR Marker in Pointed Gourd (Trichosanthes dioica Roxb.

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    Sinchan Adhikari

    2014-01-01

    Full Text Available The aim of the present study was to develop a genetic sex marker for the pointed gourd (Trichosanthes dioica Roxb. to allow gender determination at any stage in the life cycle. Screening of genomic DNA with intersimple sequence repeat (ISSR primers was used to discover sex-specific touch-down polymerase chain reaction (Td-PCR amplification products. Using pooled DNA from male and female genotypes and 42 ISSR primers, a putative male specific marker (~550 bp was identified. DNA marker specific to male is an indication of existence of nonepigenetic factors involved in gender development in pointed gourd. The ISSR technique has proved to be a reliable technique in gender determination of pointed gourd genotypes at the seedling phenophase. The sex marker developed here could also be used as a starting material towards sequence characterization of sex linked genes for better understanding the developmental as well as evolutionary pathways in sexual dimorphism.

  8. Identification of ISSR and RAPD markers linked to yield traits in bread wheat under normal and drought conditions

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    A.G.A. Khaled

    2015-12-01

    Full Text Available Genetic variability and identification of some molecular markers were studied in twenty promising lines of wheat using agronomic traits, ISSR (inter simple sequences repeats and RAPD (random amplified polymorphic DNA markers. Significant variation was evidenced in all agronomic traits. The lines proved to be superior to the check cultivar Sahel1 in yield and its component traits. Lines L2, L7 and L8 were the best in most yield component traits in both seasons. Moreover, Lines L2, L4, L5, L7 and L8 showed drought tolerance by which they displayed high performance in agronomic traits as well as a low drought susceptibility index. The percentage of polymorphism was 39.3% and 53.2% for ISSRs and RAPDs, respectively. UBC-881 belonged to penta-nucleotide repeat sequences (GGGTG that produced the highest level of polymorphism, while UBC-846 belonged to di-nucleotide repeat sequences (CA that produced the lowest level of polymorphism. Genetic similarities among wheat lines based on ISSR and RAPD markers ranged from 0.81 to 1.00 and from 0.86 to 0.98, respectively. There was a low average of PIC (polymorphism information content values which were 0.10 (ISSR and 0.15 (RAPD. The RAPD technique exhibited a higher marker index (MI = 0.69 compared to ISSR (MI = 0.43. There was insignificant correlation between ISSR and RAPD data (0.168, p > 0.05. There were two markers (UBC-881450bp and OPF-10540bp, on each of which two traits regressed significantly. The associated markers each explained a maximum regression of 18.92–34.95% of the total available variation for individual associated traits.

  9. Genetic variability analysis of Byrsonima crassifolia germplasm collected in Pará State using ISSR markers.

    Science.gov (United States)

    Rodrigues, S M; Moura, E F; Ramos, G K S; Oliveira, M S P

    2016-10-17

    Native of the Amazon, the nanche (Byrsonima crassifolia) is a fruit cultivated by family farmers and used in cooking; as such, it represents an opportunity for regional agribusiness. The Embrapa Eastern Amazon set up an active germplasm bank (BAG) consisting of 22 accessions sampled in 11 municipalities of Pará State. Due to its economic potential, there is an interest to advance the genetic breeding program of this species. The aim of this study was to characterize the BAG nanche collection using inter-simple sequence repeat (ISSR) markers. Accessions were genotyped using 23 pre-selected ISSR primers resulting in 109 amplified polymorphic and 51 monomorphic bands. With eight polymorphic bands each, the most polymorphic primers were UBC 809 and UBC 848. An unweighted pair-group method with arithmetic average cluster analysis based on Jaccard's coefficient indicated that the individuals clustered into two distinct groups. Accessions Igarapé Açu-2 and Augusto Corrêa-Pl 1 were most similar. The genetic dissimilarity values ranged from 0.10 to 0.59. We conclude that the ISSR markers were efficient in detecting polymorphisms in the nanche accessions, and that it is possible to infer the genetic variability among accessions of the collection. This demonstrate the importance of using molecular markers in poorly studied species and the advantages that this information can bring to the genetic improvement of such species.

  10. Genetic Diversity Of Plukenetia Volubilis L. Assessed By ISSR Markers*

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    Ocelák M.

    2015-12-01

    Full Text Available The diversity and genetic relationships in 173 sacha inchi samples were analyzed using ISSR markers. Thirty ISSR primers were used, only 8 showed variability in tested samples. ISSR fragments ranged from 200 to 2500 bp. The mean number of bands per primer was 12 and the average number of polymorphic bands per primer was 11. The lowest percentages of polymorphic bands (27%, gene diversity (0.103, and Shannon’s information index (0.15 were exhibited by the Santa Lucia population, which was also geographically most distant. This fact may be attributed to a very small size of this group. In contrast, the Dos de Mayo population exhibited the highest percentage of polymorphic bands (78%, and the Santa Cruz population the highest Nei’s gene diversity index (0.238 and Shannon’s information index (0.357. The obtained level of genetic variability was 36% among tested populations and 64% within populations. Although the diversity indices were low, a cluster analysis revealed 8 clusters containing mainly samples belonging to individual populations. Principal coordinate analysis clearly distinguished Chumbaquihui, Pucallpa, Dos de Mayo, and Aguas de Oro populations, the others were intermixed. The obtained results indicated the level of genetic diversity present in this location of Peru, although it is influenced by anthropological aspects and independent on the geographical distances.

  11. 玉米种质材料遗传多样性的ISSR分析%Genetic diversity of maize (Zea mays L.) accessions using inter-simple sequence repeat (ISSR) markers

    Institute of Scientific and Technical Information of China (English)

    VuVan Liet; Nguyen Thi Thuy Linh; Nguyen Thi Thuy; VuThi Bich Hanh; PhamQuangTuan; Nguyen Thi Phuong Thao

    2011-01-01

    The present study was conducted to analyze the characterization of the genetic diversity among local maize accessions in the North mountain region of Vietnam using ISSR markers.[ Method ]The ISSR technique and 10 primers were used to study genetic diversity among 21 maize accessions consisting of 12 normal and 9 waxy maize accessions,collected from 3 provinces in the North mountain region of Vietnam and Laos.[ Result]A total of 108 ISSR fragments were detected and all of them were polymorphic (100%).Polymorphism information content (PIC) values of ISSR primers ranged from 0.10-0.39.The average PIC value for each primer was 0.24.The resolving power (Rp) value ranged from 14.29-0.48 with an average of 4.48 per primer.All of tested ISSR primers with the exception of ISSR-T1 generated unique fragments (present only in one accession) in 13 among 21 maize accessions.Based on UPGMA analysis,using 70% genetic similarity as the cutoff,a dendrogram was constructed and 21 maize accessions were grouped into three clusters.The similarity coefficients among accessions ranged from 0.52-0.90.[Conclusion ]ISSR markers provided information on genetic diversity of maize accessions which was useful for collection,conservation and breeding programs in Vietnam.%[目的]利用ISSR分子标记分析越南北部山区当地玉米种质材料的遗传多样性.[方法]采用ISSR技术和10个引物对从越南和老挝北部山区3个省收集的21份玉米种质材料即12份普通玉米和9份糯玉米的遗传多样性进行分析.[结果]在21份玉米种质材料中可以检测到108个ISSR片段,其多态性为100%.ISSR引物的多态性信息量值为0.10~0.39,每条引物平均为0.24;分辨力为14.29~0.48,平均每条为4.48.除ISSR-TI以外,所有ISSR引物在13份玉米材料中均产生特异性片段.根据聚类分析结果,使用70%的遗传相似性作为切割点,建立了玉米种质材料系统树,21份玉米种质材料被分为3大类.不同玉

  12. Genetic diversity of Iranian honey bee (Apis mellifera meda Skorikow, 1829) populations based on ISSR markers.

    Science.gov (United States)

    Rahimi, A; Mirmoayedi, A; Kahrizi, D; Zarei, L; Jamali, S

    2016-04-30

    Honey bee is one of the most important insects considering its role in agriculture,ecology and economy as a whole. In this study, the genetic diversity of different Iranian honey bee populations was evaluated using inter simple sequence repeat (ISSR) markers. During May to September 2014, 108 young worker honey bees were collected from six different populations in 30 different geoclimatic locations from Golestan, Mazendaran, Guilan, West Azerbaijan, East Azerbaijan, Ardebil provinces of Iran. DNA was extracted from the worker honey bees. The quality and quantity of extracted DNA were measured. A set of ten primers were screened with the laboratory populations of honey bees. The number of fragments produced in the different honey bee populations varied from 3 to 10, varying within 150 to 1500 bp. The used ten ISSR primers generated 40 polymorphic fragments, and the average heterozygosity for each primer was 0.266. Maximum numbers of bands were recorded for primer A1. A dendrogram based on the Unweighted Pair Group Method with Arithmetic mean (UPGMA) method generated two sub-clusters. Honey bee populations of Golestan, Mazendaran, Guilan provinces were located in the first group. The second group included honey bee populations of Ardebil, West Azerbaijan, East Azerbaijan provinces, but this group showed a close relationship with other populations. The results showed obviously the ability of the ISSR marker technique to detect the genetic diversity among the honey bee populations.

  13. Applications of inter simple sequence repeat (ISSR) rDNA in ...

    African Journals Online (AJOL)

    Applications of inter simple sequence repeat (ISSR) rDNA in detecting ... and phylogenetic relationships between Lymnaea natalensis collected from Giza, ... in water samples of all tested governorates with different significant differences.

  14. Molecular characterisation and similarity relationships among iranian basil (Ocimum basilicum L. accessions using inter simple sequence repeat markers Caracterização molecular de acessos de Ocimum basilicum L. por meio de marcadores ISSR

    Directory of Open Access Journals (Sweden)

    Mohammad Aghaei

    2012-06-01

    Full Text Available The study of genetic relationships is a prerequisite for plant breeding activities as well as for conservation of genetic resources. In the present study, genetic diversity among 50 Iranian basil (Ocimum basilicum L. accessions was determined using inter simple sequence repeat (ISSR markers. Thirty-eight alleles were generated at 12 ISSR loci. The number of alleles per locus ranged from 1 to 5 with an average of 3.17. The maximum number of alleles was observed at the A7, 818, 825 and 849 loci, and their size ranged from 300 to 2500 bp. A similarity matrix based on Jaccard's coefficient for all 50 basil accessions gave values from 1.00-0.60. The maximum similarity (1.00 was observed between the "Urmia" and "Shahr-e-Rey II" accessions as well as between the "Urmia" and "Qazvin II" accessions. The lowest similarity (0.60 was observed between the "Tuyserkan I" and "Gom II" accessions. The unweighted pair- group method using arithmetique average UPGMA clustering algorithm classified the studied accessions into three distinct groups. All of the basil accessions, with the exception of "Babol III", "Ahvaz II", "Yazd II" and "Ardebil I", were placed in groups I and II. Leaf colour was a specific characteristic that influenced the clustering of Iranian basil accessions. Because of this relationship, the results of the principal coordinate analysis (PCoA approximately corresponded to those obtained through cluster analysis. Our results revealed that the geographical distribution of genotypes could not be used as a basis for crossing parents to obtain high heterosis, and therefore, it must be carried out by genetic studies.O estudo das relações genéticas é um pré-requisito para atividades em reprodução de plantas assim como para conservação de recursos genéticos. Neste trabalho a diversidade genética entre 50 acessos de Manejericão Iraniano (Ocimum basilicum L. foram determinadas usando marcadores de Seqüência Simples Repetida Interna (ISSR

  15. Analysis of Genetic Variability among thirty accessions of Andean Lupin (Lupinus mutabilis Sweet using ISSR molecular markers

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    Michelle C. Chirinos-Arias

    2015-03-01

    Full Text Available In order to make the genetic variability analysis among thirty accessions of andean lupine (L. mutabilis Sweet belonging to Agrarian Innovation National Institute (INIA Seed Bank. DNA was extracted from 300 plants and we made bulks. We standardized amplification protocol of Inter Simple Sequence Repeat (ISSR primers, we chose the most polymorphic primers to run in acrylamide gel. We found 255 ISSR loci with 8 primers. It was found high genetic variability of the samples under study by ISSR markers. Also observed relatively high polymorphism for autogamous species such as andean lupine. Finally phenograms showed a relationship with the geographical location, possibly due to in situ gene flow due to the exchange or sale of seeds in markets near the collection area.

  16. [Gene pool differentiation between Altaic and trotting horse breeds inferred from ISSR-PCR marker data].

    Science.gov (United States)

    Feofilov, A V; Bardukov, N V; Glazko, V I

    2011-09-01

    Using ISSR-PCR marker data, comparative analysis of the gene pools of Altaic and trotting horse breeds was carried out. Horse groups of different origin demonstrated differences in amplification spectra of DNA fragments flanked by inverted repeats of four microsatellites. Combinations of certain DNA fragments present in these profiles reproducibly distinguished genomes of the Altaic breed from the trotting breeds. Genetic differentiation between some trotting breeds, based on Nei genetic distance values, was found to be comparable to that between the groups of horses of Altaic breed from two different farms.

  17. Genetic diversity of Cosmos species revealed by RAPD and ISSR markers.

    Science.gov (United States)

    Rodríguez-Bernal, A; Piña-Escutia, J L; Vázquez-García, L M; Arzate-Fernández, A M

    2013-12-04

    The genus Cosmos is native of America and is constituted by 34 species; 28 of them are endemic of Mexico. The cosmos are used as a nematicide, antimalarial, and antioxidative agent. The aim of this study was to estimate the genetic diversity among 7 cosmos species based on random amplified polymorphic DNA (RAPD) and inter-simple sequences repeats (ISSR) markers. With RAPD markers, the obtained polymorphism was 91.7 % and the genetic diversity was 0.33, whereas these values were 65.6%, and 0.22 from ISSR markers, respectively, indicating the presence of high genetic diversity among the Cosmos species that were analyzed. The unweighted pair group method with arithmetic mean dendrograms that were obtained with both markers were notably similar, revealing 2 clusters and indicating a clear genetic differentiation among the Cosmos species that were assessed. The first cluster comprised the species Cosmos sulphureus, Cosmos pacificus, and Cosmos diversifolius, while the second cluster included the species Cosmos purpureus, Cosmos crithmifolius, Cosmos bipinnatus, and Cosmos parviflorus. Besides this, the Cosmos species were clustered according to their collection sites. The Mantel test corroborates the correlation between the genetic distance and the geographic altitude of each Cosmos species. The results suggest that it is necessary to preserve the Cosmos species in their natural habitat in addition to the germoplasm collection for ex situ conservation.

  18. Genetic stability of micropropagated plants of Crambe abyssinica Hochst using ISSR markers.

    Science.gov (United States)

    Werner, E T; Soares, T C B; Gontijo, A B P L; Souza Neto, J D; do Amaral, J A T

    2015-12-09

    Crambe (Crambe abyssinica) is a non-edible annual herb, which was first cultivated to extract oil for industry, and now has great potential for biodiesel production. The objective of this investigation was to evaluate the genetic stability of micropropagated plants of the C. abyssinica Hochst cultivar 'FMS brilhante' using polymerase chain reaction techniques based on inter-simple sequence repeat (ISSR) molecular markers. The aim was to develop a protocol for the in vitro regeneration of these plants with low genetic variation as compared to the donor plant. For micropropagation, shoot tips from in vitro germinated seedlings were used as explants and were initially cultivated for 90 days on MS medium with 5.0 μM 6-benzylaminopurine (BAP), which at 90 days, led to the highest number of shoots per explant (NSE) (12.20 shoots) being detected. After 120 days, the interaction between BAP concentration and naphthalene acetic acid (NAA) was tested, and the highest NSE was observed following exposure to 0.0/0.5 μM BAP/NAA (11.40 shoots) and 1.0/0.0 μM BAP/NAA (11.00 shoots). The highest proportion of rooting phase were observed following exposure to 0.5 μM NAA (30%). The 13 ISSR primers used to analyze genetic stability produced 91 amplification products, of which only eight bands were polymorphic and 83 were monomorphic for all 10 regenerated crambe plants, compared to the donor plant explant. These results indicate that crambe shoot tips are a highly reliable explant that can be used to micropropagate genetically true-to-type plants or to maintain genetic stability, as verified using ISSR markers.

  19. ISSR Marker and ITS Sequence Study of Melampsora Larici-populina

    Institute of Scientific and Technical Information of China (English)

    YU Zhong-dong; LIU Xiao-yong; CAO Zhi-min

    2006-01-01

    To compare the differences in intertranslation space of ribosomal DNA (ITS) of Melampsora larici-populina, between the isolates from China and isolates from other countries, this study investigated ITS sequences and ITS polygenetic tree based on 11 isolates that were collected from 5 races in different parts of China. The results indicated that there was no difference among the ITS sequences of 11 isolates from China. The ITS sequence of isolates from China was more homogeneous with that of isolates from Britain compared with France, Germany, and Canada. Intersimple sequence repeat(ISSR) markers were also used to study the genetic division of Melampsora larici-populina, and the results showed that the 11 tested isolates could be divided into Western population and Northern population. Genetic diversity index of race C2 was significantly different from that of races C4, C3, and C1, and no significant differences were observed among the other races. Pathogenicity division of races must not harmonize with their genetic division, except race C2. The ITS region is conservative, and ITS sequence is not fit for studying the differences that existed among the races. ISSR marker can be used for intraspecies population study, and Melampsora larici-populina in China can be divided into two populations.

  20. Genetic similarity among strawberry cultivars assessed by RAPD and ISSR markers

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    Rafael Gustavo Ferreira Morales

    2011-12-01

    Full Text Available Most strawberry (Fragaria × ananassa Duchesne cultivars used in Brazil are developed in other countries, it became clear the need to start the strawberry breeding program in the country. To start a breeding program is necessary the genetic characterization of the germplasm available. Molecular markers are important tools that can be used for this purpose. The objectives of the present study were to assess the genetic similarity among 11 strawberry cultivars using RAPD and ISSR molecular markers and to indicate the possible promising crosses. The DNA of the eleven strawberry cultivars was extracted and amplified by PCR with RAPD and ISSR primers. The DNA fragments were separated in agarose gel for the RAPD markers and in polyacrylamide gel for the ISSR markers. The genetic similarity matrix was estimated by the Jaccard coefficient. Based on this matrix, the cultivars were grouped using the UPGMA method. The dendogram generated by the RAPD markers distributed the cultivars in three groups while the ISSR markers generated two groups. There was no direct relationship between the marker groups when the two types of markers were compared. The grouping proposed by the ISSR markers was more coherent with the origin and the genealogy of the cultivars than that proposed by the RAPD markers, and it can be considered the most efficient method for the study of genetic divergence in strawberry. The most promising crosses, based on the genetic divergence estimated from the RAPD and ISSR molecular data were between the Tudla and Ventana and the Oso Grande and Ventana cultivars, respectively.

  1. Comparative Analysis of Population Genetic Structure in Bemisia tabaci (Gennadius) Biotypes B and Q Based on ISSR Marker

    Institute of Scientific and Technical Information of China (English)

    CHU Dong; WAN Fang-hao; XU Bao-yun; WU Qing-jun; ZHANG You-jun

    2008-01-01

    Bemisia tabaci (Gennadius) biotypes B and Q are two invasive biotypes in the species complex. The comparison of the population genetic structure of the two biotypes is of significance to show their invasive mechanism and to their control. The intersimple sequence repeats (ISSR) marker was used to analyze the 16 B-biotype populations and 4 Q-biotype populations worldwide with a Trialeurodes vaporariorum population in Shanxi Province, China, and a B. tabaci non-B/Q-biotype population in Zhejiang Province, China, was used as control populations. The analysis of genetic diversity showed that the diversity indexes of biotype Q including Nei's gene diversity index, Shannon informative index, and the percentage of polymorphic loci were higher than those of biotype B. The high genetic diversity of biotype Q might provide the genetic basis for the excellent ecological adaptation. Cluster analysis suggested that the ISSR could not be used in the phylogenetic analysis though it could easily distinguish the biotypes of B. tabaci. The difference of the population genetic structure between the biotype B and the biotype Q exists based on the ISSR marker. Meanwhile, the results suggested that the molecular marker has its limitation in the phylogenetic analysis among the biotypes of B. tabaci.

  2. Genetic Diversity Assessment and Identification of New Sour Cherry Genotypes Using Intersimple Sequence Repeat Markers

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    Roghayeh Najafzadeh

    2014-01-01

    Full Text Available Iran is one of the chief origins of subgenus Cerasus germplasm. In this study, the genetic variation of new Iranian sour cherries (which had such superior growth characteristics and fruit quality as to be considered for the introduction of new cultivars was investigated and identified using 23 intersimple sequence repeat (ISSR markers. Results indicated a high level of polymorphism of the genotypes based on these markers. According to these results, primers tested in this study specially ISSR-4, ISSR-6, ISSR-13, ISSR-14, ISSR-16, and ISSR-19 produced good and various levels of amplifications which can be effectively used in genetic studies of the sour cherry. The genetic similarity among genotypes showed a high diversity among the genotypes. Cluster analysis separated improved cultivars from promising Iranian genotypes, and the PCoA supported the cluster analysis results. Since the Iranian genotypes were superior to the improved cultivars and were separated from them in most groups, these genotypes can be considered as distinct genotypes for further evaluations in the framework of breeding programs and new cultivar identification in cherries. Results also confirmed that ISSR is a reliable DNA marker that can be used for exact genetic studies and in sour cherry breeding programs.

  3. ASSESSMENT OF GENETIC DIVERSITY OF REHMANNIA GLUTINOSA LIBOSCH BASED ON ISSR MARKERS

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    YANQING ZHOU, WUJUN GAO, HONGYING DUAN, FENGPING GU

    2007-08-01

    Full Text Available In order to assess the genetic diversity of Rehmannia glutinosa Libosch cultivars ( lines in Huai zone, Inter-simple sequence repeat (ISSR was performed. Ten appropriate ISSR primers were selected from a total of 44 ISSR ones for ISSR PCR amplification. The ten primers could amplify one hundred and ten bands. Based on them, A Jaccard’s genetic similarity matrix and a dendrogram for these ten cultivars were established using SPSS 10.0 software. In this dendrogram, they could be divided into two groups : Group1 contained six individuals such as Zupei 85.5, Datian 85.5, Zupei 9302, Jinbai, Jinzhuangyuan and Datian9302; Group2 consisted of four ones such as Beijing No.1, Dahongpao, Dihuang9104 and wild dihuang. Furthermore, Principal coordinate analysis (PCA supported the above cluster analysis; Shannon\\'s Information index (I is 0.3577, effective number of alleles (Ne is 1.4037, the percentage of polymorphic loci is 71.82 % by means of POPGENE32 software; A DNA fingerprint was developed with a single primer, ISSR6, in which each of ten individuals tested had its unique fingerprint pattern and was distinguished from each other. The results revealed that ISSR method is suitable for DNA fingerprinting, identification and genetic diversity analysis of Rehmannia glutinosa in Huai zone.

  4. Assessment of the Genetic Relationship and Diversity of Mango and Its Relatives by cpISSR Marker

    Institute of Scientific and Technical Information of China (English)

    HE Xin-hua; GUO Yong-ze; LI Yang-rui; OU Shi-jin

    2007-01-01

    Chloroplast inter-simple sequence repeat markers in mango were developed and used to analyze the genetic relationship and diversity of mango and its relatives. Thirty-six mango cultivars (Mangifera indica L.) and its relative species collected from the fruit germplasm collection in the Guangxi Academy of Agricultural Sciences, China, were examined by ISSR-PCR with chloroplast DNA (cpDNA). Eight better primers for chloroplast DNA that provided reproducible, polymorphic DNA amplification patterns were screened from 50 ISSR primers and used for UPGMA analysis. According to the band patterns with 8 primers for chloroplast DNA, all cultivars tested were distinguished from each other and these showed ample genetic diversity; the average percentage of polymorphism was 77.2%. The 36 samples could be clustered into four groups by UPGMA analysis at the coefficient 0.74. The results indicated that the cpISSR marker was a new powerful tool for the identification of mango cultivars or its relative species, and their genetic relationship analysis and diversity evaluation.

  5. Genetic diversity and relationships in mulberry (genus Morus as revealed by RAPD and ISSR marker assays

    Directory of Open Access Journals (Sweden)

    Thangavelu K

    2004-01-01

    Full Text Available Abstract Background The genus Morus, known as mulberry, is a dioecious and cross-pollinating plant that is the sole food for the domesticated silkworm, Bombyx mori. Traditional methods using morphological traits for classification are largely unsuccessful in establishing the diversity and relationships among different mulberry species because of environmental influence on traits of interest. As a more robust alternative, PCR based marker assays including RAPD and ISSR were employed to study the genetic diversity and interrelationships among twelve domesticated and three wild mulberry species. Results RAPD analysis using 19 random primers generated 128 discrete markers ranging from 500–3000 bp in size. One-hundred-nineteen of these were polymorphic (92%, with an average of 6.26 markers per primer. Among these were a few putative species-specific amplification products which could be useful for germplasm classification and introgression studies. The ISSR analysis employed six anchored primers, 4 of which generated 93 polymorphic markers with an average of 23.25 markers per primer. Cluster analysis of RAPD and ISSR data using the WINBOOT package to calculate the Dice coefficient resulted into two clusters, one comprising polyploid wild species and the other with domesticated (mostly diploid species. Conclusion These results suggest that RAPD and ISSR markers are useful for mulberry genetic diversity analysis and germplasm characterization, and that putative species-specific markers may be obtained which can be converted to SCARs after further studies.

  6. Genetic Similarity Assessment among Selected Naked Oat Cultivars and Breeding Lines Using ISSR Markers

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    Edyta PACZOS-GRZEDA

    2014-06-01

    Full Text Available Naked oat refers to a variety of Avena sativa with lemma and palea separating from the grains. Its spikelets are multiflorous and morphologically different from the husked oat. Problems with preharvest sprouting, threshability, rancidity, a wide range of kernel sizes, as well as its relatively low tolerance to limited soil water content, are its main drawbacks. Nevertheless it could be an alternative to a conventional oat. Unfortunately, its genetic variation is still poorly recognized.  In the given study a set of 26 naked oat cultivars and lines were analyzed with 25 inter-simple sequence repeats (ISSR primers that amplified as many as 429 DNA fragments among which 204 were polymorphic. The average number of markers amplified per primer pair and polymorphism information content (PIC value equaled to eight and 0.23, respectively. Forty four unique PCR products were identified for different genotypes. While Unweighted Pair-Group Method with Arithmetic Mean failed to distinguish the materials into main clusters it demonstrated that cultivars ‘Akt’, ‘Polar’, ‘Cacko’, ‘Siwek’, ‘Nagus’ and most of the DC lines were within a single group. Moreover, the cultivars that were closely related based on their breeding pedigree (related to ‘Akt’ were close to each other. Principal Coordinate Analysis explained 54.1% of variance and was in good agreement with the UPGMA. ISSR markers could be used for the evaluation of genetic similarity of cultivars and lines as well as the differentiation of individual genotypes. This study demonstrated that the available A. sativa naked type genetic pool is relatively wide and have the potential for further breeding progress.

  7. A comparative phylogenetic analysis of medicinal plant Tribulus terrestris in Northwest India revealed by RAPD and ISSR markers

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    ASHWANI KUMAR

    2012-07-01

    Full Text Available Kumar A, Verma N. 2012. A comparative phylogenetic analysis of medicinal plant Tribulus terrestris in Northwest India revealed by RAPD and ISSR markers. Biodiversitas 13: 107-113. Several DNA marker systems and associated techniques are available today for fingerprinting of plant varieties. A total of 5 RAPD and 8 ISSR primers were used. Amplification of genomic DNA of the 6 genotypes, using RAPD analysis, yielded 164 fragments that could be scored, of which 47 were polymorphic, with an average of 9.4 polymorphic fragments per primer. Number of amplified fragments with random primers ranged from 6 (AKR-1 to 10 (AKR-4 and varied in size from 200 bp to 2,500 bp. Percentage polymorphism ranged from 16% (AKR-4 to a maximum of 41% (AKR-4, with an average of 29.6%. The 8 ISSR primers used in the study produced 327 bands across 6 genotypes, of which 114 were polymorphic. The number of amplified bands varied from 7 (ISSR 7 to 12 (ISSR 1&3, with a size range of 250-2,800 bp. The average numbers of bands per primer and polymorphic bands per primer were 40.87 and 14.25, respectively. Percentage polymorphism ranged from 24% (ISSR 4 to 53.84% (ISSR 2, with an average percentage polymorphism of 35.59% across all the genotypes. The 3′-anchored primers based on poly (AC and poly (AT motifs produced high average polymorphisms of 53.84% and 40.81%, respectively. ISSR markers were more efficient than the RAPD assay, as they detected 35.59% polymorphic DNA markers in Tribulus terrestris as compared to 29.6% for RAPD markers. Clustering of genotypes within groups was not similar when RAPD and ISSR derived dendrogram were compared, whereas the pattern of clustering of the genotypes remained more or less the same in ISSR and combined data of RAPD and ISSR.

  8. Application of ISSR markers to analyze molecular relationships in Iranian jasmine (Jasminum spp.) accessions.

    Science.gov (United States)

    Ghasemi Ghehsareh, Masood; Salehi, Hassan; Khosh-Khui, Morteza; Niazi, Ali

    2015-01-01

    There are many species of jasmines in different regions of Iran in natural or cultivated form, and there is no information about their genetic status. Therefore, inter-simple sequence repeat (ISSR) analysis was used to evaluate genetic variations of the 53 accessions representing eight species of Jasminum collected from different regions of Iran. A total of 21 ISSR primers were used which generated 981 bands of different sizes. Mean percentage of polymorphic bands was 90.64 %. Maximum resolving power, polymorphic information content average, and marker index values were 21.55, 0.35, and 14.42 for primers of 3, 4, and 3 respectively. The unweighted pair group method with arithmetic mean dendrogram based on Jaccard's coefficients indicated that 53 accessions were divided into two major clusters. The first major cluster was divided into two subclusters; the subcluster A included Jasminum grandiflorum L., J. officinale L., and J. azoricum L. and the subcluster B consisted of three forms of J. sambac L. (single, semi-double, and double flowers). The second major cluster was divided into two subclusters; the first subcluster (C) included J. humile L., J. primulinum Hemsl., J. nudiflorum Lindl. and the second subcluster (D) consisted of J. fruticans L. At the species level, the highest percentage of polymorphism (34.05 %), numbers of effective alleles (1.16), Shannon index (0.151), and Nei's genetic diversity (0.098) were observed in J. officinale. The lowest values of percentage polymorphism (0.011), number of effective alleles (1.009), Shannon index (0.007), and Nei's genetic diversity (0.005) were obtained for J. nudiflorum. Based on pairwise population matrix of Nei's unbiased genetic identity, the highest identity (0.85) was found between J.officinale and J. azoricum and the lowest identity (0.69) was between J. grandiflorum and J. perimulinum. Based on analysis of molecular variance, the amount of genetic variations among the eight populations was 83 %. This study

  9. Identification of Phase and Sex-related ISSR Markers of Red Alga Gracilaria lemaneiformis

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Six ISSR primers are employed to display the polymorphism of different phases and sexes of red alga Gracilaria lemaneiformis, and two of them, P1 and P3, amplified distinct band patterns. The ISSR pattern amplified by primer P1 of the female gametophyte is identical to that of tetrasporophyte, but distinct from that of male gametophyte. Of the bands produced by primer P3, one is specific to female gametophyte. Three morphologically similar fronds can be easily identified using ISSR technique. Two specific markers, SM1 and SF3,related to male gametophyte and female gametophyte, are cloned and sequenced. The homologous sequences of SM1 are found to encode a hypothetical protein. There is no homologous sequence of SF3 that can be found in GenBank.

  10. Identification of phase and sex-related ISSR markers of red alga Gracilaria lemaneiformis

    Science.gov (United States)

    Xue, Sun; Xuecheng, Zhang; Yunxiang, Mao; Zhenghong, Sui; Song, Qin

    2006-01-01

    Six ISSR primers are employed to display the polymorphism of different phases and sexes of red alga Gracilaria lemaneiformis, and two of them, P1 and P3, amplified distinct band patterns. The ISSR pattern amplified by primer P1 of the female gametophyte is identical to that of tetrasporophyte, but distinct from that of male gametophyte. Of the bands produced by primer P3, one is specific to female gametophyte. Three morphologically similar fronds can be easily identified using ISSR technique. Two specific markers, SM1 and SF3, related to male gametophyte and female gametophyte, are cloned and sequenced. The homologous sequences of SM1 are found to encode a hypothetical protein. There is no homologous sequence of SF3 that can be found in GenBank.

  11. Multilocus ISSR markers reveal two major genetic groups in Spanish and South African populations of the grapevine fungal pathogen Cadophora luteo-olivacea.

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    David Gramaje

    Full Text Available Cadophora luteo-olivacea is a lesser-known fungal trunk pathogen of grapevine which has been recently isolated from vines showing decline symptoms in grape growing regions worldwide. In this study, 80 C. luteo-olivacea isolates (65 from Spain and 15 from South Africa were studied. Inter-simple-sequence repeat-polymerase chain reaction (ISSR-PCR generated 55 polymorphic loci from four ISSR primers selected from an initial screen of 13 ISSR primers. The ISSR markers revealed 40 multilocus genotypes (MLGs in the global population. Minimum spanning network analysis showed that the MLGs from South Africa clustered around the most frequent genotype, while the genotypes from Spain were distributed all across the network. Principal component analysis and dendrograms based on genetic distance and bootstrapping identified two highly differentiated genetic clusters in the Spanish and South African C. luteo-olivacea populations, with no intermediate genotypes between these clusters. Movement within the Spanish provinces may have occurred repeatedly given the frequent retrieval of the same genotype in distant locations. The results obtained in this study provide new insights into the population genetic structure of C. luteo-olivacea in Spain and highlights the need to produce healthy and quality planting material in grapevine nurseries to avoid the spread of this fungus throughout different grape growing regions.

  12. Comprehensive genetic discrimination of Leonurus cardiaca populations by AFLP, ISSR, RAPD and IRAP molecular markers.

    Science.gov (United States)

    Khadivi-Khub, Abdollah; Soorni, Aboozar

    2014-06-01

    Leonurus cardiaca is well known for its medicinal importance. In this investigation, genotypic characterization of this species from six eco-geographical regions of Iran was evaluated by four molecular techniques (AFLP, RAPD, ISSR and IRAP). A total of 899 polymorphic fragments were detected by used molecular markers (AFLP = 356, RAPD = 325, ISSR = 113 and IRAP = 105) with an overall average polymorphism of 81.24%. Genetic variation calculated using Shannon's Information index (I) and Nei's gene diversity index (H) showed high genetic diversity in studied germplasm. Also, analysis of molecular variance showed high genetic variation among (55%) and within populations (45%). UPGMA dendrogram constructed from combined data of molecular markers distinguished studied populations in accordance with the results obtained by each marker which all individuals were clearly differentiated into two major clusters. The correlation coefficients were statistically significant for all marker systems with the highest correlation between similarity matrixes of RAPD and ISSR markers (r = 0.82). The present results have an important implication for L. cardiaca germplasm characterization, improvement, and conservation. Furthermore, the characterized individuals exhibited a great deal of molecular variation and they seem to have a rich gene pool for breeding programs.

  13. Genetic diversity as assessed by ISSR markers in Blackgram (Vigna mungo (L. Hepper

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    Nadarajan N

    2009-12-01

    Full Text Available An investigation was carried out on a collection of 23 blackgram genotypes involving 16 releasedvarieties, six pre release cultures and one wild species Vigna mungo var. silvestris to study the genetic diversityusing twelve ISSR primers. The number of alleles produced by different ISSR primers ranged from eight to 17with an average of 11.5 per primer and the level of polymorphism was found to be 82.05 percent. Similaritymeasures and clustering analyses were made using ISSR data. The resulting dendrogram distributed the 23blackgram genotypes into five main clusters. The highest genetic similarity coefficient was measured betweengenotypes CBG 671 and CBG 632. The results of PCoA were comparable to that of grouping based on UPGMAand 23 genotypes were grouped into four groups. Genotype Vigna mungo var. silvestris was placed separatelyfrom rest of the genotypes in both the analyses. Grouping of varieties using ISSR markers did not show anyrelevance to their pedigree. All the pre release cultures in one group revealed that only a portion of geneticvariation has been exploited. The results revealed that, genetic diversity is low among the varieties releasedfrom the respective institute and hence genotypes were grouped according to the research institutes from whichthey released. It suggests that the research institutes have to enlarge the genetic base for variety development.

  14. Development of simple sequence repeats (SSR) markers of ramie and comparison of SSR and inter-SSR marker systems

    Institute of Scientific and Technical Information of China (English)

    ZHOU Jianlin; JIE Yucheng; JIANG Yanbo; ZHONG Yingli; LIU Yunhai; ZHANG Jian

    2005-01-01

    Ramie (Boehmeria nivea L. ) is an important bast fiber crop. To study genetic background of this species, we isolated and characterized microsatellite markers of ramie. A genomic library containing inserts of rapid amplification of polymorphic DNA (RAPD)fragments was constructed, and screened by PCR amplification using anchored simple sequence repeats as primers. A total of 26 clones were identified as positives, and 13 microsatellite loci were found after sequencing. The polymorphism of these 13 microsatellite loci was examined and the utility of simple sequence repeats (SSR) and inter-SSR (ISSR) marker systems for genetic characterization compared using 19 selected ramie cultivars. Both approaches successfully discriminated the 19 cultivars which differed in the amount of polymorphism detected. The level of polymorphism detected by SSR was 95.0 %, higher than that by ISSR (72.3 % ), but the average polymorphism information content (PIC) of ISSR (0. 651) was higher than that of SSR (0. 441). The higher PIC value of ISSR suggests that ISSR is more efficient for fingerprinting ramie cultivars than SSR markers. However, because the SSR loci are codominant, they are more suitable for determining the homozygosity levels of ramie, constructing linkage map, quantitative trait loci study of complex traits and marker-as-sisted selection.

  15. Assessment of genetic diversity in tomato landraces using ISSR markers

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    Henareh Mashhid

    2016-01-01

    Full Text Available Tomato is one of the most economically important vegetable crops in many parts of the world. Turkey and Iran are the main producers of tomatoes in the world. The objective of this study was to assess the genetic variation of 93 tomato landraces from East Anatolian region of Turkey and North-West of Iran, along with three commercial cultivars using 14 ISSR primers. The percentage of polymorphic loci (PPL for all primers was 100%. The mean of expected heterozygosity (He for the primers varied from 0.153 (UBC808 to 0.30 (UBC848. The dendrogram placed the landraces and commercial cultivars into nine groups. The genotypes originating from the same region, often located in the same group or two adjacent groups. The highest likelihood of the data was obtained when population were located into 2 sub-populations (K = 2. These sub-populations had Fst value of 0.16 and 0.21.

  16. Molecular characterization of Prunus mahaleb L. rootstock canditates by ISSR markers

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    Ozyurt Ibrahim Kursat

    2013-01-01

    Full Text Available Prunus mahaleb is widely used as rootstocks particularly on calcareous and dry soils for both sweet and sour cherry cultivars in Turkey. Genetic diversity and relationships among members of Prunus mahaleb including 29 preselected rootstock candidate accessions from Tokat region in Turkey were investigated by using 15 ISSR markers. The study revealed high genetic diversity among accessions, detecting 138 fragments, of which 103 (75% were polymorphic. The number of polymorphic bands per primer was between 3-13, with average of 6.86. The primers 890 and 891 gave the highest polymorphism ratio (100%. The UPGMA dendrogram and the principal coordinate analysis revealed a clear differentiation among accessions. Reference rootstock, SL-64 clustered separately. The study demonstrates that ISSRs provide promising marker tools in revealing genetic diversity and relationships in Prunus mahaleb rootstock candidate accessions and can contribute to efficient identification, conservation, and utilization of germplasm for rootstock improvement through conventional as well as molecular breeding approaches.

  17. The use of ISSR markers for species determination and a genetic study of the invasive lionfish in Guanahacabibes, Cuba

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    Elizabeth Labastida

    2015-11-01

    Full Text Available The red lionfish (Pterois volitans and devil fire-fish (Pterois miles are invasive species that pose a threat to the biodiversity and stability of coral reefs in the Western Atlantic, Gulf of Mexico and Caribbean Sea. Species identification of lionfish is uncertain in some parts of Cuba, and research has mainly been focused on their biology and ecology. The principal aim of this study was to determine highly polymorphic markers (Inter Simple Sequence Repeat, ISSR that could be used in research on lionfish population genetics in addition to confirming the presence of Pterois species in the Guanahacabibes National Park. The genetic profile or "fingerprint" of individuals collected in Mexico, formally identified as P. volitans, was compared with the genetic profile of specimens from Cuba. There were very few "diagnostic bands" and a high number of "common bands", demonstrating that the same species exists in both countries. Furthermore, Nei's genetic distance and the unrooted tree do not show significant differences between both localities. In light of these results, we can confirm the presence of P. volitans in the Guanahacabibes National Park, Cuba. This study demonstrates the functionality of ISSR as a molecular tool for species identification and their application for genetic population studies of this invasive fish species.

  18. Molecular diversity and phylogeny of Triticum-Aegilops species possessing D genome revealed by SSR and ISSR markers

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    Moradkhani Hoda

    2015-12-01

    Full Text Available The aim of this study is investigation the applicability of SSR and ISSR markers in evaluating the genetic relationships in twenty accessions of Aegilops and Triticum species with D genome in different ploidy levels. Totally, 119 bands and 46 alleles were detected using ten primers for ISSR and SSR markers, respectively. Polymorphism Information Content values for all primers ranged from 0.345 to 0.375 with an average of 0.367 for SSR, and varied from 0.29 to 0.44 with the average 0.37 for ISSR marker. Analysis of molecular variance (AMOVA revealed that 81% (ISSR and 84% (SSR of variability was partitioned among individuals within populations. Comparing the genetic diversity of Aegilops and Triticum accessions, based on genetic parameters, shows that genetic variation of Ae. crassa and Ae. tauschii species are higher than other species, especially in terms of Nei’s gene diversity. Cluster analysis, based on both markers, separated total accessions in three groups. However, classification based on SSR marker data was not conformed to classification according to ISSR marker data. Principal co-ordinate analysis (PCoA for SSR and ISSR data showed that, the first two components clarified 53.48% and 49.91% of the total variation, respectively. This analysis (PCoA, also, indicated consistent patterns of genetic relationships for ISSR data sets, however, the grouping of accessions was not completely accorded to their own geographical origins. Consequently, a high level of genetic diversity was revealed from the accessions sampled from different eco-geographical regions of Iran.

  19. Molecular analysis of commercial date palm cultivars in Lybia using ISSR and SRAP PCR-based markers

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    Khalifa Noha S.

    2016-01-01

    Full Text Available Little is known about the molecular structure of the date palm (Phoenix dactylifera L. despite its importance as invaluable drought tolerant crop. Intervarietal variation and cultivar identification are crucial for breeding and gene bank conservation of this plant worldwide. In this work, two PCR based marker systems (ISSR and SRAP were applied on top quality eight commercial cultivars in Libya (Umfetity, Bekrary, Alhamraya, Sufeer Genab, Alsaeedy Show, Farag Barameel, Majhool Alheelo and Alkhadraya. DNA variations were explored using eleven ISSR and nine combinations of SRAP markers. All markers used generated polymorphic bands among the different cultivars that can be used as molecular markers for their differentiation. The genetic distance between cultivars was also estimated from banding patterns. Our results indicate that ISSR and SRAP systems can efficiently identify and differentiate between the selected cultivars. This work can be used as a model to establish a road map for all date palm cultivars worldwide.

  20. CHARACTERIZATION OF A NEW BIOTYPE Moringa OF SAUDI ARABIA USING RAPD AND ISSR MARKERS

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    Iyan Robiansyah

    2015-11-01

    Full Text Available Moringa peregrina and M. oleifera are the only Moringa species found in Saudi Arabia. Both species are drought resistant and have very high nutritional and medicinal properties. Detection of genetic diversity is of great value for the improvement of nutritional and medicinal value of these plants. The aim of the present study was to characterize a new biotype Moringa observed in Al Bahah Region, Saudi Arabia. We used 11 RAPD and 15 ISSR primers to characterize and compare the new biotype with M. peregrina and M. oleifera. Level of polymorphism generated by each marker was calculated. We also calculate Nei and Li’s coefficient to measure the genetic distance between the studied species. Level of polymorphism generated by RAPD and ISSR was 46% and 57%, respectively. RAPD and ISSR primers revealed that the new biotype shared 55 amplicons (45.08% with both M. peregrina and M. oleifera, 28 amplicons with M. peregrina (22.95%, 21 amplicons (17.21% with M. oleifera, and displayed 18 unshared amplicons (14.75%. Based on RAPD data, genetic distance between M. oleifera and M. peregrina was 0.32, whereas genetic distance between the new biotype and M. oleifera and M. peregrina was 0.21 and 0.29, respectively. For ISSR data, genetic distance between M. oleifera and M. peregrina was 0.5, whereas genetic distance between the new biotype and M. oleifera and M. peregrina was 0.36 and 0.34, respectively. Based on these results we suggested that the new biotype is a hybrid crossbred between M. peregrina and M. oleifera.

  1. Micropropagation and validation of genetic and biochemical fidelity among regenerants of Nothapodytes nimmoniana (Graham) Mabb. employing ISSR markers and HPLC.

    Science.gov (United States)

    Prakash, Lokesh; Middha, Sushil Kumar; Mohanty, Sudipta Kumar; Swamy, Mallappa Kumara

    2016-12-01

    An in vitro protocol has been established for clonal propagation of Nothapodytes nimmoniana which is an important source of Camptothecin (CPT). Elite source was identified based on the chemical potency to accumulate the optimum level of CPT. Different types and concentrations of plant growth regulators were used to study their effect on inducing multiple shoots from the explants regenerated from embryos of N. nimmoniana. Of these, a combination of N6-benzyladenine (0.2 mg L(-1)) and Indole-3-butyric acid (IBA) (0.1 mg L(-1)) proved optimum for differentiating multiple shoots in 90.6 % of the cultures with an average of 10.24 shoots per explant obtained within 8 weeks of inoculation. Nearly, 92 % of the excised in vitro shoots rooted on half strength Murashige and Skoog (MS) medium containing 0.05 % activated charcoal, supplemented with 1-naphthaleneacetic acid and IBA at 0.1 mg L(-1) each. The micropropagated plants were evaluated for their genetic fidelity by employing inter simple sequence repeats (ISSR) markers. Ten individuals, randomly chosen from a population of 145 regenerants, were compared with the donor plant. The regenerated plants were also evaluated for their chemical potency using high-performance liquid chromatography (HPLC) analysis of CPT content. The true-to-type nature of the micropropagated plants was confirmed based on their monomorphic banding profiles with that of the mother plants using ISSR markers. Besides, HPLC evaluation of the CPT content confirmed the existence of chemical uniformity among the regenerated plants and the elite mother plant.

  2. Genetic similarity between coriander genotypes using ISSR markers Similaridade genética entre genótipos de coentro por marcadores ISSR

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    Roberto de A Melo

    2011-12-01

    Full Text Available With the development of new cultivars, a precise genetic characterization is essential for improvement programs or for cultivar registration and protection. Molecular markers have been complementing the traditional morphological and agronomic characterization techniques because they are virtually unlimited, cover the whole genome and are not environmentally influenced. Genetic characterization constitutes the basis for studies involving estimates of genetic similarity. Therefore, the objective of the present study was to evaluate the genetic similarity between ten coriander genotypes (nine cultivars and one line using ISSR markers. The cultivars used were: Americano, Asteca, Palmeira, Português, Santo, Supéria, Tabocas, Tapacurá, Verdão and the experimental line HTV-9299. The genetic similarity between the cultivars was estimated using 227 banded regions of ISSR molecular markers. The UBC 897 oligonucleotide generated the highest number of fragments (16, resulting in a higher polymorphism. The results indicate that the twenty-nine oligonucleotides chosen were satisfactory for detecting polymorphism. Based on the grouping analysis determined from the similarity data, there were two groups and two sub-groups. The calculated similarity for the genotypes varied from 52 to 75%. The lowest similarity was observed between Português and Verdão, at 52%. The highest similarity was found between Português and Palmeira, at 75%. The ISSR is efficient for identifying DNA polymorphism in coriander.Com o surgimento de novas cultivares, uma caracterização genética precisa é essencial, visando à utilização em programas de melhoramento ou para fins de registros e ou proteção de cultivares. Marcadores moleculares vêm complementando a caracterização morfológica e agronômica tradicional, uma vez que são virtualmente ilimitados, cobrem todo o genoma e não são influenciados pelo ambiente. A caracterização genética constitui a base para

  3. ISSR markers for analysis of molecular diversity and genetic structure of Indian teak (Tectona grandis L.f. populations

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    Shamin Akhtar Ansari

    2012-06-01

    Full Text Available Inter simple sequence repeats (ISSR constitute a powerful dominant DNA molecular marker system used for diversity analysis, which is indispensable for making estimates of genetic base and demarcation of populations for undertaking conservation and improvement program offorest tree species. Twenty nine populations of teak (Tectona grandis L.f. were collected from central and peninsular India for analysis of genetic diversity and structure. Genomic DNA from ten randomly selected individuals of each population was extracted and amplified using five ISSR primers (UBC-801, 834, 880, 899 and 900. The primers showed 100% polymorphism. UBC-900 recorded the highest Nei's genetic diversity (0.32 to 0.40 and UBC-899 had the highest Shannon's Information Index (0.49 to 0.59. AMOVA revealed a very high intra-population genetic diversity (91%, in comparison to inter-population genetic diversity among states (6.17% and within states (2.77%, were also indirectly confirmed by large standard deviations associated with genetic diversity estimates for individual population, as well as poor bootstrapping values for most of the cluster nodes. However, UPGMA dendrogram revealed several clusters, with populations from central India being present almost in each cluster, making groups with populations of adjoining states and distant states. Nevertheless, the cluster analysis distinguished the drier teak populations of central India from the moist teak populations of south India, which was also confirmed by Principle Coordinate Analysis. The findings advocates the need not only for enhancing selection intensity for large number of plus trees, but also for laying out more number of in situ conservation plots within natural populations of each cluster for germplasm conservation of teak aimed at improving the teak productivity and quality in future. 

  4. ISSR markers for analysis of molecular diversity and genetic structure of Indian teak (Tectona grandis L.f. populations

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    S.A. Ansari

    2012-05-01

    Full Text Available Inter simple sequence repeats (ISSR constitute a powerful dominantDNA molecular marker system used for diversity analysis, which isindispensable for making estimates of genetic base and demarcation of populations for undertaking conservation and improvement program of forest tree species. Twenty nine populations of teak (Tectona grandis L.f. were collected from central and peninsular India for analysis of genetic diversity and structure. Genomic DNA from ten randomly selected individuals of each population was extracted and amplified using five ISSR primers(UBC-801, 834, 880, 899 and 900. The primers showed 100% polymorphism. UBC-900 recorded the highest Nei’s genetic diversity (0.32 to 0.40and UBC-899 had the highest Shannon’s Information Index (0.49 to 0.59. AMOVA revealed a very high intra-population genetic diversity (91%, in comparison to inter-population genetic diversity among states (6.17% and within states (2.77% which were also indirectly confirmed by large standard deviations associated with genetic diversity estimates for individual population, as well as poor bootstrapping values for most of the cluster nodes. However, UPGMA dendrogram revealed several clusters, with populationsfrom central India being present almost in each cluster, makinggroups with populations of adjoining states and distant states. Nevertheless,the cluster analysis distinguished the drier teak populations of central India from the moist teak populations of south India, which was also confirmed by Principle Coordinate Analysis. The findings advocates the need not only for enhancing selection intensity for large number of plus trees, but also for laying out more number of in situ conservation plots within natural populations of each cluster for germplasm conservation of teak aimed at improving the teak productivity and quality in future.

  5. Genetic variability in apomictic mangosteen (Garcinia mangostana and its close relatives (Garcinia spp. based on ISSR markers

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    SOBIR

    2011-04-01

    Full Text Available Sobir, Poerwanto R, Santosa E, Sinaga S, Mansyah E (2011 Genetic variability in apomictic mangosteen (Garcinia mangostana and its close relatives (Garcinia spp. based on ISSR markers. Biodiversitas 12: 59-63. In order to reveal phylogenetic relationship of mangosteen and several close relatives (Garcinia spp., we employed seven ISSR dinucleotide primer systems on eleven close relatives of mangosteen and 28 mangosteen accessions from four islands in Indonesia (Sumatra, Java, Kalimantan and Lombok. ISSR analysis successfully amplified 43 bands on average 6.1 fragments for each primer system, and these all fragments were polymorphic. Seven close relatives of mangosteen were separated with mangosteen accessions at 0.22 level of dissimilarity, while other four including G. malaccensis, were clustered with mangosteen accessions, this results supported proposal that G. malaccensis was allopolyploid derivative of mangosteen. Clustering pattern among mangosteen accessions, however, not represented their origin, indicated that distribution of the accessions was not linked to their genetic properties.

  6. Molecular characterization of arabica and Conilon coffee plants genotypes by SSR and ISSR markers

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    Ludymila Brandão Motta

    2014-10-01

    Full Text Available The molecular characterization of ten genotypes of the Coffea arabica plants and of seven genotypes of C. canephora having interesting features for coffee breeding programs was carried to select the parents for breeding. A total of 40 SSR and 29 ISSR primers were used. The primers generated a total of 331 (307 polymorphic and 24 monomorphic bands. Analysis of genetic diversity presented dissimilarity intervals ranging from 0.22 to 0.44 between the Conilon genotypes, from 0.02 to 0.28 between the Arabica genotypes, and from 0.49 to 0.60 between the genotypes of the two species in the joint analysis. Four groups were formed: I = genotypes of C. arabica, II = four progenies of C. canephora, Conilon group, and one non defined C. canephora (Conilon or Robusta, III = one progeny of un-defined C. canephora (Conilon or Robusta and IV = one progeny of C. canephora of Robusta group. The grouping formed was consistent with the origins of each group. High stabilities of the bifurcations were found by bootstrap analysis. The use of molecular markers of the SSR and ISSR types in the diversity study was efficient in distinguishing genotypes between and within C. arabica and C. canephora.

  7. A study of conservation genetics in Cupressus chengiana, an endangered endemic of China, using ISSR markers.

    Science.gov (United States)

    Hao, Bingqing; Li, Wang; Linchun, Mu; Li, Yao; Rui, Zhang; Mingxia, Tang; Weikai, Bao

    2006-02-01

    ISSR markers were used to analyze the genetic diversity and genetic structure of eight natural populations of Cupressus chengiana in China. ISSR analysis using 10 primers was carried out on 92 different samples. At the species level, 136 polymorphic loci were detected. The percentage of polymorphic bands (PPB) was 99%. Genetic diversity (He) was 0.3120, effective number of alleles (Ae) was 1.5236, and Shannon's information index (I) was 0.4740. At the population level, PPB = 48%, Ae = 1.2774, He = 0.1631, and I = 0.2452. Genetic differentiation (Gst) detected by Nei's genetic diversity analysis suggested 48% occurred among populations. The partitioning of molecular variance by AMOVA analysis indicated significant genetic differentiation within populations (54%) and among populations (46%; P < 0.0003). The average number of individuals exchanged between populations per generation (Nm) was 0.5436. Samples from the same population clustered in the same population-specific cluster, and two groups of Sichuan and Gansu populations were distinguishable. A significantly positive correlation between genetic and geographic distance was detected (r = 0.6701). Human impacts were considered one of the main factors to cause the rarity of C. chengiana, and conservation strategies are suggested based on the genetic characters and field investigation, e.g., protection of wild populations, reestablishment of germplasm bank, and reintroduction of more genetic diversity.

  8. Genetic Diversity Assessment and Identification of New Sour Cherry Genotypes Using Intersimple Sequence Repeat Markers

    OpenAIRE

    Roghayeh Najafzadeh; Kazem Arzani; Naser Bouzari; Ali Saei

    2014-01-01

    Iran is one of the chief origins of subgenus Cerasus germplasm. In this study, the genetic variation of new Iranian sour cherries (which had such superior growth characteristics and fruit quality as to be considered for the introduction of new cultivars) was investigated and identified using 23 intersimple sequence repeat (ISSR) markers. Results indicated a high level of polymorphism of the genotypes based on these markers. According to these results, primers tested in this study specially IS...

  9. Effects of genotoxicity and its consequences at the population level in sexual and asexual Artemia assessed by analysis of inter-simple sequence repeats (ISSR).

    Science.gov (United States)

    Sukumaran, Sandhya; Grant, Alastair

    2013-09-18

    There is considerable evidence that genetic damage in organisms occurs in the environment as a result of exposure to genotoxins and ionising radiation, but we have limited understanding of the extent to which this results in adverse consequences at a population level. We used inter-simple sequence repeat (ISSR) markers to quantify genotoxic effects of the mutagen ethylmethane sulfonate (EMS) on a sexual (Artemia franciscana) and an asexual (Artemia parthenogenetica) species of brine shrimp. The method provides information similar to that obtained with assessment of RAPD (random amplification of polymorphic DNA) but is more robust. Genetic damage was transmitted to the F1 generation in both Artemia species, but the sexual species showed a greater degree of recovery, as shown by higher values of genomic template stability. There was a strong correlation between DNA damage and effects on individual fitness parameters: size, survival, reproduction and population growth. These effects persisted into the F2 generation in A. parthenogenetica, but in the sexual A. franciscana only effects on fecundity continued beyond the exposed generation, even though there were substantial alterations in ISSR patterns in the F1 generation. Genetic biomarkers can thus be indicative of effects at the population level, but sexually reproducing species have a considerable assimilative capacity for the effects of genotoxins.

  10. Assessment of genetic diversity in the Russian olive (Elaeagnus angustifolia based on ISSR genetic markers Avaliação da diversidade genética em Oliva Russa (Elaeagnus angustifolia com base em marcadores genéticos ISSR

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    Leila Sadat Asadiar

    2013-06-01

    Full Text Available Elaeagnus is a Eurasian tree with 77 species worldwide. In this study, ISSR markers were used to establish the level of genetic relationships and polymorphism across nine genotypes of Elaeagnus angustifolia collected from 9 different regions of West Azarbaijan province. The ISSR analysis with 11 anchored primers also generated 116 scorable loci, of which 92 were polymorphic (79.3%. The estimated Jaccard similarity coefficient ranged from 0.44 to 0.76 for the ISSR markers. Cluster analysis was carried out, based on the Unweighted Pair Group Method with Arithmetic Averages (UPGMA and the dendrogram drawn with the help of the NTSYSpc 2.02 software. The analysis revealed 5 main clusters for the ISSR data. According to our results, there is a relatively high genetic distance across E. angustifolia genotypes in the West Azarbaijan province of Iran. Furthermore, it could be inferred that ISSR markers are suitable tools for the evaluation of genetic diversity and relationships within the Elaeagnus genus.A Elaeagnus é uma árvore da Eurásia com 77 espécies em todo o mundo. Neste estudo, marcadores ISSR foram usados para estabelecer o nível de relações genéticas e polimorfismo entre nove genótipos de Elaeagnus angustifolia, coletados em 9 diferentes regiões da província do Azerbaijão Ocidental. A análise ISSR, com 11 primers ancorados, também gerou 116 loci contáveis, dos quais 92 polimórficos (79,3%. O coeficiente de similaridade de Jaccard estimado, variou de 0,44 a 0,76 para os marcadores ISSR. A análise de agrupamento foi realizada com base no Método não-ponderado de pares não-agrupados, com médias aritméticas (UPGMA, e a dendrograma elaborada com a ajuda do software NTSYSpc 2.02. A análise revelou cinco grupos principais para os dados ISSR. De acordo com nossos resultados, há uma distância genética relativamente alta entre genótipos de E. angustifolia na província de Azarbaijan Ocidental no Iran. Além disso, pode

  11. Identification of 'Ubá' mango tree zygotic and nucellar seedlings using ISSR markers

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    Aline Rocha

    2014-10-01

    Full Text Available Polyembryonic seeds are characterized by the development of over one embryo in the same seed, which can be zygotic and nucellar. The objective of this work was to identify the genetic origin, whether zygotic or nucellar, of seedlings of polyembryonic seeds of 'Ubá' mango tree using ISSR markers, and relating them with the vigor of the seedlings. Thus, mangos were harvested in Visconde do Rio Branco (accession 102 and Ubá (accessions 112, 138, 152 and 159, whose seeds were germinated in plastic trays filled with washed sand. Fifty days after sowing, seedlings from five seeds of each one of the accessions 102, 112, 138, 159 and from 10 seeds of the accession 152, were analyzed. These sseedlings were characterized and evaluated for plant height, stem circumference and mass of fresh aerial part and the most vigorous seedling was the one displaying at least two of these traits higher than the other seedlings from seed. Leaves were collected for genomic DNA extraction, which was amplified using seven ISSR primers previously selected based on the amplification profile and considering the number and resolution of fragments. Zygotic seedlings were found in 18 seeds, which were the most vigorous in six seeds. The results evidenced the existence of genetic variability in orchards using seedlings grown from seeds, because the farmer usually uses the most vigorous ones, assuming that this is of nucellar origin. These results also indicate that the most vigorous seedling are not always nucellar, inasmuch as of 20% of the total seeds evaluated, the zygotic seedling was the most vigorous.

  12. Micropropagation and assessment of genetic fidelity of Dendrocalamus strictus (Roxb.) nees using RAPD and ISSR markers.

    Science.gov (United States)

    Goyal, Arvind Kumar; Pradhan, Sushen; Basistha, Bharat Chandra; Sen, Arnab

    2015-08-01

    Dendrocalamus strictus popularly known as 'Male bamboo' is a multipurpose bamboo which is extensively utilized in pharmaceutical, paper, agricultural and other industrial implements. In this study, in vitro regeneration of D. strictus through nodal culture has been attempted. Murashige and Skoog's medium supplemented with 4 mg/l BAP was found to be most effective in shoot regeneration with 3.68 ± 0.37 shoots per explant. The effect of Kn was found to be moderate. These hormones also had considerable effect on the shoot length. The highest shoot length after 6 weeks (3.11 ± 0.41 cm) was noted with 5 mg/l BAP followed by 3.07 ± 0.28 cm with 5 mg/l Kn, while decrease in the shoot length was noted with other treatments. The effect of IBA and NAA individually or in combination at different concentrations on rooting was evaluated. The highest number of root (1.36 ± 0.04) was regenerated on full-strength MS medium supplemented with 3 mg/l NAA, while maximum length of 1.64 ± 0.03 cm of roots was recorded with combination of 1 mg/l IBA and 3 mg/l NAA. Tissue-cultured plants thus obtained were successfully transferred to the soil. The clonal fidelity among the in vitro-regenerated plantlets was assessed by RAPD and ISSR markers. The ten RAPD decamers produced 58 amplicons, while nine ISSR primers generated a total of 66 bands. All the bands generated were monomorphic. These results confirmed the clonal fidelity of the tissue culture-raised D. strictus plantlets and corroborated the fact that nodal culture is perhaps the safest mode for multiplication of true to type plants.

  13. Characterization of new variety of Chrysanthemum by using ISSR markers Caracterização de novas cultivares de crisântemo com o uso de marcadores ISSR

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    SK Palai

    2011-12-01

    Full Text Available Chrysanthemum is the important cut flower after rose among the ornamental plants traded in the global flower market. It is propagated vegetatively and also has a strong sporophytic self-incompatibility system as shown by all members of Asteraceae family. Morphologically, the petal numbers and flower colours present maximum variation when compared to existing varieties. Twenty Inter Simple Sequence Repeat primers were used to detect the new variety of Chrysanthemum developed through spontaneous sporting. The results indicate that the rate of polymorphism showed significant differences as compared to other existing varieties. The average number of amplification products per primer was eight. The size of ISSR amplified fragments varied from 0.25 - 2.4 Kbp. Therefore, ISSR marker is a useful technique for the rapid and easy assessment of genetic variation among the variants. Morphological traits of new variants showed variation as compared to other parents. The 1st flower bud appearance and the height of 1st bud of the variant were less as compared to original mother variety. The new variants can be propagated in large scale commercially through in vitro technique.Entre as plantas ornamentais comercializados no mercado mundial, o crisântemo é a flor de corte de maior importância sendo superado apenas pela rosa. Ele é propagado vegetativamente e também tem um forte sistema de auto-incompatibilidade esporofítica como mostrado por todos os membros da família Asteraceae. Morfologicamente, os números de pétalas e as cores das flores apresentam variação máxima em relação às cultivares existentes. Empregou-se vinte primers ISSR (Inter Simple Sequence Repeat para caracterizar a nova cultivar de crisântemo desenvolvida por mutação expontânea. Os resultados indicam que a taxa de polimorfismo mostrou diferenças significativas em comparação com outras cultivares existentes. Foi de oito o número médio de produtos de amplificação por

  14. The loss of genetic diversity during captive breeding of the endangered sculpin, Trachidermus fasciatus, based on ISSR markers: implications for its conservation

    Institute of Scientific and Technical Information of China (English)

    BI Xiaoxiao; YANG Qiaoli; GAO Tianxiang; LI Chuangju

    2011-01-01

    Inter-simple sequence repeat (ISSR) markers were used to determine the genetic variation and genetic differentiation of cultured and wild populations of Trachidermus fasciatus,an endangered catadromous fish species in China.Six selected primers were used to amplify DNA samples from 85 individuals,and 353 loci were detected.Relatively low genetic diversity was detected in the cultured population (the percentage of polymorphic loci PPL=73.80%,Nei's gene diversity h=0.178 2,Shannon information index I=0.276 9).However,the genetic diversity at the species level was relatively high (PPL=91.78%; h=0.258 3,I=0.398 6).The UPGMA tree grouped together the genotypes almost according to their cultured and wild origin,showing distinct differences in genetic structure between wild and cultured populations.The pairwise Fst values confirmed significant genetic differentiation between wild and cultured samples.The cultivated population seems to be low in genetic diversity as a result of detrimental genetic effects in the captive population.The results suggest that ISSR markers are effective for rapid assessment of the degree of diversity of a population,thus giving important topical information relevant to preserving endangered species.

  15. The Application of ISSR Markers to Identify the Fertility Restorer Gene Rf6 in T.timopheevii Cytoplasmic Male Sterile Wheat(Triticum aestivum)

    Institute of Scientific and Technical Information of China (English)

    GUAN Rong-xia; GUO Xiao-li; LIU Dong-cheng; CAO Shuang-he; ZHANG Ai-min

    2002-01-01

    Inter-simple sequence repeat (ISSR) analysis was carried out on a F2 population of 147 plants derived from a cross between a wheat male fertility restorer line 2114 and a male sterile line ND44A. Out of 43 primers examined, 18 primers produced distinguishable, polymorphic bands between the two parents. Linkage analysis in the mapping population showed that two markers UBC-808 and UBC-848 were closely linked with the restorer gene R f6 of the Triticum timopheevii CMS system. The distance between the two markers and the restorer gene was 7.9 cM and 4.9 cM, respectively. Also two parents were screened with 181 pairs of SSR primers, of which, 34.3% showed polymorphisms. But no locus was found linked with the restorer gene.Compared with the SSR technique, the ISSR approach used in the experiment provided more information and proved to be a valuable method to identify alien fragments.

  16. Analysis of genetic diversity of Brassica rapa var. chinensis using ISSR markers and development of SCAR marker specific for Fragrant Bok Choy, a product of geographic indication.

    Science.gov (United States)

    Shen, X L; Zhang, Y M; Xue, J Y; Li, M M; Lin, Y B; Sun, X Q; Hang, Y Y

    2016-04-25

    Non-heading Chinese cabbage [Brassica rapa var. chinensis (Linnaeus) Kitamura] is a popular vegetable and is also used as a medicinal plant in traditional Chinese medicine. Fragrant Bok Choy is a unique accession of non-heading Chinese cabbage and a product of geographic indication certified by the Ministry of Agriculture of China, which is noted for its rich aromatic flavor. However, transitional and overlapping morphological traits can make it difficult to distinguish this accession from other non-heading Chinese cabbages. This study aimed to develop a molecular method for efficient identification of Fragrant Bok Choy. Genetic diversity analysis, based on inter-simple sequence repeat molecular markers, was conducted for 11 non-heading Chinese cabbage accessions grown in the Yangtze River Delta region. Genetic similarity coefficients between the 11 accessions ranged from 0.5455 to 0.8961, and the genetic distance ranged from 0.0755 to 0.4475. Cluster analysis divided the 11 accessions into two major groups. The primer ISSR-840 amplified a fragment specific for Fragrant Bok Choy. A pair of specific sequence-characterized amplified region (SCAR) primers based on this fragment amplified a target band in Fragrant Bok Choy individuals, but no band was detected in individuals of other accessions. In conclusion, this study has developed an efficient strategy for authentication of Fragrant Bok Choy. The SCAR marker described here will facilitate the conservation and utilization of this unique non-heading Chinese cabbage germplasm resource.

  17. Discrimination Capacity of RAPD, ISSR and SSR Markers and of their Effectiveness in Establishing Genetic Relationship and Diversity among Egyptian and Saudi Wheat Cultivars

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    Salah E.D. El-Assal

    2012-01-01

    Full Text Available Problem statement: Yield crop cultivars and landraces are valuable sources of genetic variations that the knowledge and implication of these variations are critical in the plant breeding programs. our major objective of this study is investigating the discriminating capacity of RAPD, ISSR and SSR markers and of their effectiveness in establishing genetic relationship and diversity among Egyptian and Saudi wheat cultivars. Approach: Eleven wheat cultivars and landraces collected from Egypt and Saudi Arabia, five Egyptian wheat (Sakha 93, Sods 1, Sods 4, Gmiza 9 and Sohag 3 and six Saudi wheat landrace cultivars (Hmees, Al-Kaseem, Hegazi, Abo-Sakr, Dubai 1 and Nagran were characterized using RAPD, ISSR and SSR molecular markers as efficient tools. Ten and nine oligonucleotide primers of RAPD and ISSR respectively and four primer pairs of SSR were used in wheat samples analysis. Only clear and repeatable band profile of 6 RAPD, 8 ISSR and 2 SSR primers were obtained. In RAPD analyses, 74 out of 141 bands (52% were polymorphic. Results: The number of alleles ranged from 8-21 per primer, with an average of 14.1 per primer. In ISSR analyses, a total of 78 alleles were detected, along with 36 alleles (46% were polymorphic. The number of alleles per primer ranged from 5-10 with an average of 8.6 alleles per ISSR primer. SSR reactions recorded 6 alleles, of which 5 alleles (83% were polymorphic. Cluster analysis was conducted using Unweighted Pair Group Method that depends on Arithmetic Average (UPGMA. The dendrogram cluster diagram classified the evaluated genotypes in three major clusters corresponding to the cultivation regions. The first group contains Sakha 93, Sods 1 and Sods 4 with more than 80% Genetic Similarity (GS. The GS between Sakha 93 and Sods 1, Sakha 93 and Sods 4 or Sods 1 and Sods 4 were 83.6%, 83.9 and 85.4 respectively. The second group contains Gmiza 9 and Sohag 3 with GS 83.1%. The third group contains most of the Saudi landrace

  18. Assessment of genetic diversity and population structure of an endemic Moroccan tree (Argania spinosa L.) based in IRAP and ISSR markers and implications for conservation.

    Science.gov (United States)

    Pakhrou, Ouafae; Medraoui, Leila; Yatrib, Chaimaa; Alami, Mohammed; Filali-Maltouf, Abdelkarim; Belkadi, Bouchra

    2017-07-01

    Argan Tree is well known for its precious oil extracted from its seeds particularly used for the nutritional and cosmetic benefits. Because of the high international demand, the argan tree suffers from overexploitation and its cultivation is rare. Thus, the assessment of the genetic variation of this endemic tree is critically important for designing conservation strategies. In the present study and for the first time, genetic diversity of the global natural distribution of argan tree (Argania spinosa L.) in Morocco was assessed. Four IRAP (inter-retrotransposon amplified polymorphism) primer combinations and seven ISSR (inter-simple sequence repeat) primers amplified 164 and 248 scorable polymorphic bands respectively. Polymorphic information content (PIC = 0.27), resolving power (Rp = 15) and marker index (MI = 10.81) generated by IRAP primer combinations were almost identical to those generated by ISSR primers (PIC = 0.27, Rp = 9.16 and MI = 12). AMOVA analysis showed that 49% of the genetic variation was partitioned within populations which is supported by Nei's genetic differentiation (Gst = 0.5391) and the overall estimate of gene flow (Nm) being 0.4274. The STRUCTURE analysis, PCoA (principal coordinate analysis) and UPGMA (unweighted pair-group method with arithmetic mean) based on the combined data matrices of IRAP and ISSR divided the 240 argan genotypes into two groups. The strong differentiation observed might be due to the geographical distribution of argan tree. Our results provide crucial insight for genetic conservation programs of this genetic resource.

  19. Genetic variation within three populations of Phycella australis (Phil. Ravenna from Biobío Region, Chile, evaluated using ISSR markers

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    Cristian Flores

    2013-03-01

    Full Text Available Phycella australis (Phil. Ravenna is a Chilean plant with high ornamental potential; however, the intensive extraction as a cut flower might be detrimental for the conservational state by ignoring the state of genetic variation. The objective of this investigation was to assess genetic variability within and between three populations of P. australis in the Biobío Region using inter-simple sequence repeat (ISSR markers. The evaluated areas correspond to three locations in the province of Concepción, Biobío Region: Desembocadura (36°48' S, 73°10' W, Santa Juana (36°58' S, 72°58' W, and Lipinhue (37°00' S, 72°58' W. Six ISSR primers were used obtaining 51 fragments, from which 72.5% were polymorphic. From the three evaluated sites Santa Juana showed a higher percentage of polymorphic loci (76.47%. From this variability, 83% belong to within population variability and only 17% belong to variability between populations. The dendrogram generated using the unweighted pair group method with arithmetic mean (UPGMA method, grouped Lipinhue and Santa Juana sites together, which agrees with the geographic locations. This investigation proved that P. australis has high genetic variability despite the exploitation for economic purposes.

  20. Study of genetic variation in sesame (Sesamum indicum L.) using agro-morphological traits and ISSR markers.

    Science.gov (United States)

    Parsaeian, M; Mirlohi, A; Saeidi, G

    2011-03-01

    This research was conducted to study the genetic variation among eighteen genotypes of sesame (Sesamum indicum L.) collected from various agro-climatic regions of Iran along with six exotic genotypes from the Asian countries using both agro-morphological and ISSR marker traits. The results showed significant differences among genotypes for all agro-morphological traits and a relatively high genetic coefficient of variation observed for number of fruiting branches per plant, capsules per plant, plant height and seed yield per plant. Cluster analysis based on these traits grouped the genotypes into five separate clusters. Larger inter- than intra cluster distances implies the presence of higher genetic variability between the genotypes of different groups. Genotypes of two clusters with a good amount of genetic divergence and desirable agronomic traits were detected as promising genotypes for hybridization programs. The 13 ISSR primers chosen for molecular analysis revealed 170 bands, of which 130 (76.47%) were polymorphic. The generated dendrogram based on ISSR profiles divided the genotypes into seven groups. A principal coordinate analysis confirmed the results of clustering. The agro-morphological traits and ISSR markers reflected different aspects of genetic variation among the genotypes as revealed by a non significant cophenetic correlation in the Mantel test. Therefore the complementary application of both types of information is recommended to maximize the efficiency of sesame breeding programs. The discordance among diversity patterns and geographical distribution of genotypes found in this investigation implies that the parental lines for hybridization should be selected based on genetic diversity rather than the geographical distribution.

  1. Assessment of genetic diversity in Trigonella foenum-graecum and Trigonella caerulea using ISSR and RAPD markers

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    Ranjekar Prabhakar K

    2004-07-01

    Full Text Available Abstract Background Various species of genus Trigonella are important from medical and culinary aspect. Among these, Trigonella foenum-graecum is commonly grown as a vegetable. This anti-diabetic herb can lower blood glucose and cholesterol levels. Another species, Trigonella caerulea is used as food in the form of young seedlings. This herb is also used in cheese making. However, little is known about the genetic variation present in these species. In this report we describe the use of ISSR and RAPD markers to study genetic diversity in both, Trigonella foenum-graecum and Trigonella caerulea. Results Seventeen accessions of Trigonella foenum-graecum and nine accessions of Trigonella caerulea representing various countries were analyzed using ISSR and RAPD markers. Genetic diversity parameters (average number of alleles per polymorphic locus, percent polymorphism, average heterozygosity and marker index were calculated for ISSR, RAPD and ISSR+RAPD approaches in both the species. Dendrograms were constructed using UPGMA algorithm based on the similarity index values for both Trigonella foenum-graecum and Trigonella caerulea. The UPGMA analysis showed that plants from different geographical regions were distributed in different groups in both the species. In Trigonella foenum-graecum accessions from Pakistan and Afghanistan were grouped together in one cluster but accessions from India and Nepal were grouped together in another cluster. However, in both the species accessions from Turkey did not group together and fell in different clusters. Conclusions Based on genetic similarity indices, higher diversity was observed in Trigonella caerulea as compared to Trigonella foenum-graecum. The genetic similarity matrices generated by ISSR and RAPD markers in both species were highly correlated (r = 0.78 at p = 0.001 for Trigonella foenum-graecum and r = 0.98 at p = 0.001 for Trigonella caerulea indicating congruence between these two systems

  2. Genetic and chemical diversity of high mucilaginous plants of Sida complex by ISSR markers and chemical fingerprinting.

    Science.gov (United States)

    Thul, Sanjog T; Srivastava, Ankit K; Singh, Subhash C; Shanker, Karuna

    2011-09-01

    A method was developed based on multiple approaches wherein DNA and chemical analysis was carried out toward differentiation of important species of Sida complex that is being used for commercial preparation. Isolated DNA samples were successfully performed through PCR amplification using ISSR markers and degree of genetic diversity among the different species of Sida is compared with that of chemical diversity. For genetic fingerprint investigation, selected 10 ISSR primers generating reproducible banding patterns were used. Among the total of 63 amplicons, 62 were recorded as polymorphic, genetic similarity index deduced from ISSR profiles ranged from 12 to 51%. Based on similarity index, S. acuta and S. rhombifolia found to be most similar (51%). High number of species-specific bands played pivotal role to delineate species at genetic level. Investigation based on HPTLC fingerprints analysis revealed 23 bands representing to characteristic chemicals and similarity index ranged from 73 to 91%. Prominent distinguishable bands were observed only in S. acuta, while S. cordifolia and S. rhombifolia shared most bands making them difficult to identify on chemical fingerprint basis. This report summarizes the genotypic and chemotypic diversity and the use of profiles for authentication of species of Sida complex.

  3. Genetic Diversity of Chinese and Swedish Rapeseed (Brassica napus L. ) Analyzed by Inter-Simple Sequence Repeats (ISSRs)

    Institute of Scientific and Technical Information of China (English)

    MA Chao-zhi; FU Ting-dong; Stine Tuevesson; Bo Gertsson

    2003-01-01

    We have compared genetic diversity of 24 Chinese weak-winter, Swedish winter and spring B.napus accessions by inter-simple sequence repeats (ISSRs). By cluster analysis (UPGMA) based on 125 polymorphism bands amplified with 20 primers, the 24 accessions were divided into three groups. Six Swedish winter lines and eight Chinese weak-winter lines were in the group Ⅰ and the group Ⅱ were two Chinese weakwinter lines Xiangyou15 and Bao81. The third group contained eight Swedish spring lines. Principal co-ordinates analysis (PCO) showed similar groupings to cluster analysis. Results from cluster analysis and PCO analysis showed very clearly that Chinese weak-winter, Swedish spring and winter accessions were distinguished from each other and Chinese weak-winter accessions in this study were genetically closer to Swedish winter accessions than to Swedish spring accessions. The Chinese weak-winter accessions had larger diversity than Swedish spring or winter accessions did. This study indicated that ISSR is a suitable and effective tool to evaluate genetic diversity among rapeseed germplasm.

  4. In Vitro Mass Multiplication and Assessment of Genetic Stability of In Vitro Raised Artemisia absinthium L. Plants Using ISSR and SSAP Molecular Markers

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    B. Kour

    2014-01-01

    Full Text Available The present investigations were made attempting to develop a rapid, reliable, and reproducible in vitro regeneration protocol for Artemisia absinthium L., a medicinal plant of Kashmir Himalayas. Out of several auxin-cytokinin combinations tested, Murashige and Skoog’s (MS medium supplemented with 0.5 mgL−1 2,4-dichlorophenoxyacetic acid (2,4-D and 0.5 mgL−1 kinetin (Kn was found to be the best for the callus induction. On the other hand, 4.5 mgL−1 6-benzylaminopurine (BAP and 0.5 mgL−1 1-α-naphthaleneacetic acid (NAA in the medium resulted in maximum shoot induction from the callus. Similarly, BAP and NAA at a concentration of 1.5 mgL−1 and 0.5 mgL−1, respectively, proved to be the best for the multiple shoot induction from nodal explants. Numerous shoots were obtained from nodal explants after third subculture. In vitro rooting was maximum on medium containing indole-3-butyric acid (IBA at 0.5 mgL−1. The genetic stability of the in vitro raised plants of Artemisia absinthium was assessed using the intersimple sequence repeat (ISSR and sequence-specific amplification polymorphism (SSAP molecular markers. Both markers were able to detect the somaclonal variations in the callus regenerated plants, while no variation was detected in the plants regenerated from the nodal explants. SSAP has been found to be more useful in detection of variability as compared to ISSR molecular marker. The results of present study concluded that the direct regeneration protocol will be useful for the production of true to type plants of this medicinally important plant. This will go a long way in reducing the pressure on the natural populations for the secondary metabolite production, especially for extraction of essential oils.

  5. Construction of a linkage map and QTL analysis of horticultural traits for watermelon [Citrullus lanatus (THUNB.) MATSUM & NAKAI] using RAPD, RFLP and ISSR markers.

    Science.gov (United States)

    Hashizume, T; Shimamoto, I; Hirai, M

    2003-03-01

    We have been constructing linkage maps for watermelon ( Citrullus lanatus) on the basis of random amplified polymorphic DNA (RAPD), restriction fragment length polymorphism (RFLP), inter-simple sequence repeats (ISSRs) and isozymes using an F(2) population derived from a crossing between a cultivated inbred line (H-7; C. lanatus) and an African wild form (SA-1; C. lanatus). A total of 120 F(2) plants was used for construction of a linkage map using 477 RAPDs, 53 RFLPs, 23 ISSRs and one isozyme markers. Linkage analysis revealed that 554 loci could be mapped to 11 linkage groups that extended for 2,384 centimorgans (cM). While a BC(1) population [(H-7 x SA-1) x H-7] consisting of 60 individuals was grown and scored for quantitative traits. Another linkage map with a total length of 1,729 cM was constructed in the BC(1) using genetic markers found to segregate in the F(2) population. A QTL analysis was applied by means of interval mapping for locating such agronomic traits as hardness of rind, Brix of flesh juice, flesh color (red and yellow) and rind color. The relative order of markers in the BC(1) map was essentially the same as that on the linkage map in the F(2). A total of five QTLs for four agronomic traits was detected. The QTL for hardness of rind was mapped on group 4. The linkage group 8 contained the QTL for sugar content of the flesh as expressed in Brix of the juice. The QTL for red flesh color was detected on groups 2 and 8. The QTL for rind color mapped on the group 3. The present map and QTL analysis may provide a useful tool for breeders by introducing valuable wild watermelon genes to cultivars.

  6. Screening for eri silkworm (Samia ricini Donovan ecoraces using morphological characters, growth, yields, and ISSR marker

    Directory of Open Access Journals (Sweden)

    Duanpen Wongsorn

    2015-10-01

    Full Text Available The selection of eri silkworm ecoraces with high yield and distinct morphological characters is necessary for variety improvement. The five ecoraces SaKKU1, SaKKU2, SaKKU3, SaKKU4 and SaKKU5 were derived mostly by international academic cooperation. They were cultured using castor leaves of TCO 101 cultivar as food plant at 25±2°C, 80±5% R.H. Based on morphological characters, they are similar, except the body of the 5th instar larva of SaKKU1 is clearly covered with more creamy white powder and the mature larva has a shiny dominant yellow color. The duration of the life cycle among ecoraces was also similar; 46-53 days (SaKKU1, 42-53 days (SaKKU2, 42-52 days (SaKKU3, 40-56 days (SaKKU4 and 41-52 days (SaKKU5. SaKKU1 had the highest survival rate at larval stage (1st – 5th instar (100.00% and larva (1st – 5th instar - adult (88.89%, including the predominant heaviest average larva weight of all instars, 0.0317 g (2nd instar, 0.2206 g (3rd instar, 1.0788 g (4th instar, 4.0102 g (5th instar, and 8.9940 g (5 days of 5th instar, which was significantly different (P<0.05 to other ecoraces. Moreover, this ecorace gave the highest average yields: fresh cocoon weight (3.8016 g, pupa weight (3.2532 g, shell weight (0.5287 g, shell ratio (14.01%, fresh cocoon weight/10,000 larvae (38.01 kg, eggs/moth (531.13 eggs, total eggs (6,375.27 eggs and total hatching eggs (6,006.13 eggs, which was also significantly different (P<0.05 than other ecoraces. Of those properties, especially survival rates and yields, this ecorace (SaKKU1 is favored for further varietal improvement program. In parallel, genetic relationship analysis of eri silkworm ecoraces using inter-simple sequence repeat (ISSR technique was also carried out. The result revealed from dendrogram analysis that SaKKU1 was the farthest distance than other ecoraces, especially against SaKKU3. Based on all above results, the SaKKU1 ecorace was considered to be the most suitable for heat tolerant

  7. Use of ISSR markers to assess the genetic diversity in wild medicinal Ziziphus spina-christi (L. Willd. collected from different regions of Saudi Arabia

    Directory of Open Access Journals (Sweden)

    Saleh Alansi

    2016-09-01

    Full Text Available Ziziphus spina-christi (sidr is a shrub, sometimes a tree, native to a vast area of Africa stretching from Mauritania to West Africa. In the Kingdom of Saudi Arabia, it is an exotic medicinal plant for many diseases. The aim of this study was to assess the genetic diversity within and among 34 accessions of Z. spina-christi collected from different regions of Saudi Arabia. The amplification of genomic DNA with 11 inter-simple sequence repeat (ISSR primers yielded 105 scorable loci, of which 93.4% were found to be polymorphic. The observed number of alleles (na, effective number of alleles (ne, Nei's gene diversity (h and genetic diversity estimated by Shannon's information index (I were 1.93, 1.44, 0.26 and 0.41, respectively. The total genetic diversity, Ht (0.266 ± 0.0289 was close to the average intrapopulation genetic diversity, Hs (0.2199 ± 0.0216. A high level of gene flow (Nm = 2.37 between populations, reflecting high genetic differentiation (Gst = 0.1739. The analysis of molecular variance showed that the maximum value of genetic variation was found within populations (90%, whereas a low value of genetic variance was observed among populations. The analysis using the unweighted pair-group method with arithmetic averages clustered the population from Farasan Island as an out-group due to its geographical origin. The obtained results demonstrate that the ISSR markers may be used for evaluation of the genetic diversity due to their efficiency in revealing polymorphism even in closely related germplasm and may help in Ziziphus genome analysis.

  8. Genetic Homogeneity Revealed Using SCoT, ISSR and RAPD Markers in Micropropagated Pittosporum eriocarpum Royle- An Endemic and Endangered Medicinal Plant.

    Science.gov (United States)

    Thakur, Julie; Dwivedi, Mayank D; Sourabh, Pragya; Uniyal, Prem L; Pandey, Arun K

    2016-01-01

    Pittosporum eriocarpum Royle, a medicinally important taxon, is endemic to Uttarakhand region of Himalaya. It has become endangered due to over-collection and the loss of habitats. As raising plants through seeds in this plant is problematic, a reliable protocol for micropropagation using nodal explants has been developed. High shoot regeneration (95%) occurred in MS medium augmented with BA 0.4mg/l in combination IBA 0.6mg/l. In vitro regenerated shoots were rooted in MS medium supplemented with three auxins, of which 0.6 mg/l indole butyric acid proved to be the best for rooting (90%) with maximum number of roots per shoot. Thereafter, rooted plants were hardened and nearly 73% of rooted shoots were successfully acclimatized and established in the field. Start codon targeted (SCoT), inter simple sequence repeats (ISSR) and random amplified polymorphic DNA (RAPD) markers were used to validate the genetic homogeneity amongst nine in vitro raised plantlets with mother plant. DNA fingerprints of in vitro regenerated plantlets displayed monomorphic bands similar to mother plant, indicating homogeneity among the micropropagated plants with donor mother plant. The similarity values were calculated based on SCoT, ISSR and RAPD profiles which ranged from 0.89 to 1.00, 0.91 to 1.00 and 0.95 to 1.00 respectively. The dendrograms generated through Unweighted Pair Group Method with arithmetic mean (UPGMA) analysis revealed 97% similarity amongst micropropagated plants with donor mother plant, thus confirming genetic homogeneity of micropropagated clones. This is the first report on micropropagation and genetic homogeneity assessment of P. eriocarpum. The protocol would be useful for the conservation and large scale production of P. eriocarpum to meet the demand for medicinal formulations and also for the re-introduction of in vitro grown plants in the suitable natural habitats to restore the populations.

  9. Genetic Homogeneity Revealed Using SCoT, ISSR and RAPD Markers in Micropropagated Pittosporum eriocarpum Royle- An Endemic and Endangered Medicinal Plant

    Science.gov (United States)

    Thakur, Julie; Dwivedi, Mayank D.; Sourabh, Pragya; Uniyal, Prem L.; Pandey, Arun K.

    2016-01-01

    Pittosporum eriocarpum Royle, a medicinally important taxon, is endemic to Uttarakhand region of Himalaya. It has become endangered due to over-collection and the loss of habitats. As raising plants through seeds in this plant is problematic, a reliable protocol for micropropagation using nodal explants has been developed. High shoot regeneration (95%) occurred in MS medium augmented with BA 0.4mg/l in combination IBA 0.6mg/l. In vitro regenerated shoots were rooted in MS medium supplemented with three auxins, of which 0.6 mg/l indole butyric acid proved to be the best for rooting (90%) with maximum number of roots per shoot. Thereafter, rooted plants were hardened and nearly 73% of rooted shoots were successfully acclimatized and established in the field. Start codon targeted (SCoT), inter simple sequence repeats (ISSR) and random amplified polymorphic DNA (RAPD) markers were used to validate the genetic homogeneity amongst nine in vitro raised plantlets with mother plant. DNA fingerprints of in vitro regenerated plantlets displayed monomorphic bands similar to mother plant, indicating homogeneity among the micropropagated plants with donor mother plant. The similarity values were calculated based on SCoT, ISSR and RAPD profiles which ranged from 0.89 to 1.00, 0.91 to 1.00 and 0.95 to 1.00 respectively. The dendrograms generated through Unweighted Pair Group Method with arithmetic mean (UPGMA) analysis revealed 97% similarity amongst micropropagated plants with donor mother plant, thus confirming genetic homogeneity of micropropagated clones. This is the first report on micropropagation and genetic homogeneity assessment of P. eriocarpum. The protocol would be useful for the conservation and large scale production of P. eriocarpum to meet the demand for medicinal formulations and also for the re-introduction of in vitro grown plants in the suitable natural habitats to restore the populations. PMID:27434060

  10. RAPD and ISSR Methods Used for Fingerprinting of Selected Accessions of Viburnum

    Directory of Open Access Journals (Sweden)

    Marcelina KRUPA-MAŁKIEWICZ

    2014-09-01

    Full Text Available Randomly amplified polymorphic DNA (RAPD and inter simple sequence repeat (ISSR markers were used to investigate genetic variability within thirteen Viburnum species (Viburnum × hillieri; V. dilatatum; Viburnum × carlcephalum; V. opulus; V. hupehense; Viburnum× bodnantense; Viburnum × burkwoodii; V. sieboldii; Viburnum × globosum ‘Jermyns Globe’; V. alnifolium (lantanoides; V. plicatum ‘Sterile’; V. plicatum f. tomentosum and V. plicatum ‘Watanabe’ of wide geographical distribution, collected in the Dendrological Garden in Przelewice (the north-west part of Poland. Twenty-three RAPD and fourteen ISSR primers generated a total of 690 and 418 reproducible bands, respectively, and 39% (RAPD and 55.5% (ISSR of them were polymorphic for the two marker systems, which suggest high genetic variability within Viburnum genus. However, high numbers of genotype-specific bands, i.e. 60.9% (RAPD and 44.5% (ISSR, were seen in Viburnum. Genetic similarity assessed within Viburnum species with the RAPD and ISSR analyses ranged from 6 to 42% and from 6 to 31%, respectively. Both RAPD and ISSR-based dendrograms clustered in five main groups. The Mantel test between two Nei’s similarity matrices gave correlation coefficient r=0.305*, showing low correlation between RAPD- and ISSR- based matrices. Thus, both marker systems were equally important for the genetic diversity analysis in Viburnum genus.

  11. Genetic fidelity and variability of micropropagated cassava plants (Manihot esculenta Crantz) evaluated using ISSR markers.

    Science.gov (United States)

    Vidal, Á M; Vieira, L J; Ferreira, C F; Souza, F V D; Souza, A S; Ledo, C A S

    2015-07-14

    Molecular markers are efficient for assessing the genetic fidelity of various species of plants after in vitro culture. In this study, we evaluated the genetic fidelity and variability of micropropagated cassava plants (Manihot esculenta Crantz) using inter-simple sequence repeat markers. Twenty-two cassava accessions from the Embrapa Cassava & Fruits Germplasm Bank were used. For each accession, DNA was extracted from a plant maintained in the field and from 3 plants grown in vitro. For DNA amplification, 27 inter-simple sequence repeat primers were used, of which 24 generated 175 bands; 100 of those bands were polymorphic and were used to study genetic variability among accessions of cassava plants maintained in the field. Based on the genetic distance matrix calculated using the arithmetic complement of the Jaccard's index, genotypes were clustered using the unweighted pair group method using arithmetic averages. The number of bands per primer was 2-13, with an average of 7.3. For most micropropagated accessions, the fidelity study showed no genetic variation between plants of the same accessions maintained in the field and those maintained in vitro, confirming the high genetic fidelity of the micropropagated plants. However, genetic variability was observed among different accessions grown in the field, and clustering based on the dissimilarity matrix revealed 7 groups. Inter-simple sequence repeat markers were efficient for detecting the genetic homogeneity of cassava plants derived from meristem culture, demonstrating the reliability of this propagation system.

  12. Application and Prospect of Inter-simple Sequence Repeat (ISSR) in Silkworm Genetics%ISSR分子标记技术在家蚕遗传研究上的应用进展

    Institute of Scientific and Technical Information of China (English)

    吴凡; 李德臣; 赵春晓; 陈登松

    2015-01-01

    The basic principle and characteristics of Inter-simple Sequence Repeat (ISSR) was described. The application of ISSR in silkworm genetics was summarized. The application future in the study of silkworm was prospected.%概括了ISSR 分子标记技术的基本原理和特点,综述了其在家蚕相关领域中应用的研究进展,并展望了其在家蚕遗传研究上的应用前景。

  13. Regeneration and assessment of genetic fidelity of the endangered tree Moringa peregrina (Forsk.) Fiori using Inter Simple Sequence Repeat (ISSR).

    Science.gov (United States)

    Al Khateeb, Wesam; Bahar, Eman; Lahham, Jamil; Schroeder, Dana; Hussein, Emad

    2013-01-01

    Moringa peregrinais an endangered species of Moringaceae.M. peregrinais a multipurpose tree with a wide variety of potential uses including its medicinal activity. In our study, a rapid and efficient micropropagation protocol for M. peregrina has been established. In vitro germinated seedlings were cultured on Murashige and Skoog (MS) medium supplemented with different levels of either 6-benzyladenine (BA) or kinetin (Kin). The maximum shoot proliferation of 6.5 shoots per explant with 100 % shoot proliferation rate was observed on MS medium supplemented with 1.0 mg/l BA. On the other hand, MS medium supplemented with 1 mg/l indole-3-butyric acid (IBA) resulted in the maximum number of roots. Micropropagated plants were successfully acclimatized. Genetic stability of micropropagated plants was assessed using Inter-Simple Sequence Repeat (ISSR). The amplification products were monomorphic in all in vitro grown plants. No polymorphism was detected indicating the genetic integrity of in vitro propagated plants. This micropropagation protocol could be useful for raising genetically uniform plants for plant propagation and commercial cultivation.

  14. 采用ISSR-PCR分子标记法鉴别黑果枸杞与雄性不育枸杞%Identification of Lycium ruthenicum Murr and Male Sterility of Lycium barbarum L. by Inter-Simple Sequence Repeat Markers

    Institute of Scientific and Technical Information of China (English)

    思彬彬; 张靠稳; 刘国迪

    2012-01-01

    [Objective] To identify Lycium ruthenicum and male sterility of Lycium barbarum by inter-simple sequence repeat markers. [ Method] Primers suitable for routine analysis were screened from 100 inter-simple sequence repeat primers, then, they were used in PCR and separated of 2 samples of L. Ruthenicum and male sterility of /- barbarum L. [ Results] Four of the 100 primers amplified polymorphic bands and suitable for the identification of L ruthenicum and male sterility oft. Barbarum. [Conclusion] Inter-simple sequence repeat markers provide a quick, reliable molecular marker for identification of L ruthenicum and male sterility of L barbarum.%[目的]探索用ISSR分子标记方法在核酸分子水平上鉴别黑果枸杞与雄性不育枸杞.[方法]从100条ISSR引物中筛选合适的引物对黑果枸杞与雄性不育枸杞样品进行PCR扩增及电泳分析,寻找特征位点.[结果]有4条ISSR引物扩增出较为明显的多态性特征条带,可单独应用于黑果枸杞与雄性不育枸杞的鉴别.[结论]ISSR作为一种简便、可靠的分子标记方法,可用于黑果枸杞与雄性不育枸杞的鉴别.

  15. ISSR-PCR技术在对虾中的应用初步研究%Preliminary study on the application of inter-simple sequence repeats (ISSR)-PCR technique in Chinese shrimp (Fenneropenaeus chinensis)

    Institute of Scientific and Technical Information of China (English)

    王伟继; 孔杰

    2002-01-01

    采用ISSR(Inter Simple Sequence Repeats)技术对中国对虾(Penaeus chinensis)进行了PCR扩增.优化了PCR反应体系和反应参数,对PCR产物进行聚丙烯酰胺凝胶电泳和银染检测.从100条ISSR引物中筛选了40条有清晰产物的引物,每条引物检测到的位点数从1到19不等,平均每条引物可检测到位点数为7.7个.实验发现:中国对虾简单重复序列区主要由两碱基循环组成.通过分析ISSR-PCR技术本身的原理,探讨了该技术相对于同工酶检测和RAPD技术在遗传多样性分析中的优势所在,以及该技术用于遗传标识的确认和遗传图谱构建方面的前景.

  16. Genetic Diversity Analysis of Morinda citrifolia by ISSR Markers%海巴戟ISSR遗传多样性分析

    Institute of Scientific and Technical Information of China (English)

    廖丹; 程江波; 聂风琴; 李明; 林道哲; 符文英

    2016-01-01

    The genetic diversity and cluster analysis of the 202 Morinda citrifolia germplasm resources were an-alyzed by using ISSR molecular markers.The results showed that 13 ISSR primers which amplified 65 loci,meaned the average of each primer could amplified five loci.Among them,63 bands (96.92%)were polymorphic.Cluster analysis result indicated that 202 germplasm resources could be clustered into five groups when the genetic similari-ty coefficient was 0.63,revealedthe characteristics of geographical distribution.The average of Shannon information index (I),Neis genetic diversity (H)and effective number of alleles (Ne)was 0.5206,0.3488 and 1.6042, respectively,which indicated there was highly genetic diversity in these 202 Morinda citrifolia resource .%利用ISSR分子标记对202份海巴戟种质材料进行遗传多样性和聚类分析。结果表明:13条ISSR引物共扩增出65个位点,每个引物平均扩增出5个位点,其中多态性位点63个,占总数的96.92%。在相似系数为0.63时,202份海巴戟种质分为5个类,表现出一定的地域分布规律。检测到有效等位基因数 Ne =1.6042,基因多样性 H=0.3488,Shannon′s信息指数 I=0.5206,说明202份海巴戟种质间存在着较高的遗传多样性。

  17. Optimization of Inter-Simple Sequence Repeats(ISSR) Reaction System for Agaricus blazei%姬松茸ISSR特异扩增体系的研究

    Institute of Scientific and Technical Information of China (English)

    林新坚; 江秀红; 蔡海松; 郑永标; 林陈强

    2007-01-01

    为了建立稳定的姬松茸(Agaricus blazei Murill)简单序列重复区间(Inter-Simple Sequence Repeats,ISSR)分子标记技术体系,笔者通过单因子试验分别研究了模板DNA、Mg2+浓度、dNTP、引物浓度和Taq酶用量对姬松茸ISSR-PCR扩增的影响,确定了姬松茸ISSR分析的最佳PCR条件为:25μL反应体系中,模板DNA 20 ng,引物0.75μmol/L,dNTP 200μmol/L,Mg2+2.0 mmol/L,Taq DNA polymerase 1.5 U.并应用该优化体系筛选到6个适合姬松茸ISSR-PCR扩增的引物,为利用ISSR标记技术研究姬松茸的种质资源提供了参考.

  18. [Clonal micropropagation of a rare species Hedysarum theinum Krasnob (Fabaceae) and assessment of the genetic stability of regenerated plants using ISSR markers].

    Science.gov (United States)

    Erst, A A; Svyagina, N S; Novikova, T I; Dorogina, O V

    2015-02-01

    In the present study, a protocol was developed for the in vitro propagation of a rare medicinal plant, Hedysarum theinum (tea sweetvetch), from axillary buds, and identification of the regenerants was performed with the use of ISSR markers. It was demonstrated that Gamborg and Eveleigh medium supplemented with 5 μM 6-benzylaminopurine was the best for H. theinum for initial multiplication. On the other hand, half-strength Murashige and Skoog (MS) basal medium supplemented with 7 μM α-naphthaleneacetic acid proved to be the best for explant rooting. Molecular genetic analysis of the H. theinum mother plants and the obtained regenerants was performed with six ISSR markers. Depending on the primer, four to ten amplified fragments with sizes ranging from 250 to 3000 bp were identified. Our results confirmed the genetic stability of regenerants obtained in five passages and their identity to the mother plant.

  19. 法国蜜环菌ISSR-PCR反应体系的优化%Optimization of inter-simple sequence repeat PCR (ISSR-PCR) reaction system for Armillaria gallica

    Institute of Scientific and Technical Information of China (English)

    孙立夫; 张艳华; 裴克全; 杨国亭; 宋玉双; 秦国夫; 宋瑞清

    2009-01-01

    本文用单因子试验分析了基于ISSR(Inter-Simple Sequence Repeat)分子标记研究法国蜜环菌系统发生学的PCR(Polymerase Chain Reaction)扩增反应条件,并进行了引物筛选,同时对各个引物的退火温度以及甲酰胺对扩增效果的影响进行了讨论.为利用ISSR标记技术研究蜜环菌生物种的系统发生学、遗传多样性及种质资源提供了参考.

  20. Analysis of genetic diversity of Laeliinae (Orchidaceae) in the State of Sergipe using ISSR markers.

    Science.gov (United States)

    Arrigoni-Blank, M F; Santos, M S; Blank, A F; Rabbani, A R C; Silva-Mann, R; Santos, J B; Costa, A S; Menezes, T S A

    2016-06-03

    The Orchidaceae represent one of the largest and most diverse families on the planet. However, this family is constantly threatened by predators and by the advancement of urban centers over its natural habitats. The objective of this study was to use inter-simple sequence repeat markers to evaluate the genetic diversity between orchid accessions of the Laeliinae subtribe, which comprise part of the Orchidaceae study collection at the Department of Agronomic Engineering of the Federal University of Sergipe. DNA was extracted from each specimen by using an adapted 2% cetyltrimethyl ammonium bromide protocol. Similarity between individuals was calculated using the Jaccard method. Clustering was carried out by the unweighted pair group method with arithmetic mean method, with resampling and 10,000 bootstraps. Eighty-seven fragments were obtained, all of which were polymorphic, revealing high variability between accessions. The mean similarity was 35.77% between Encyclia sp individuals, and 35.90% between specimens of Cattleya tigrina. For Epidendrum secundum, a relationship between geographic and genetic distances was observed, and the accession collected in the southern part of the State of Sergipe (Serra de Itabaiana National Park) was more divergent than that of the other parts of the state. The data generated in this study will guide further research aimed at the ex situ conservation of these materials.

  1. Genotoxic effect of Pb and Cd on in vitro cultures of Sphagnum palustre: An evaluation by ISSR markers.

    Science.gov (United States)

    Sorrentino, Maria Cristina; Capozzi, Fiore; Giordano, Simonetta; Spagnuolo, Valeria

    2017-08-01

    In the present work, the genotoxic effect of cadmium and lead supplied in a laboratory trial, was investigated for the first time in the moss Sphagnum palustre, by ISSR molecular markers. A total of 169 reproducible bands were obtained with 12 primers, ten of which gave polymorphisms (i.e., appearance/disappearance of bands), indicating a clear genotoxic effect induced by the metals. Both metals induced a decrease of the genome template stability in a dose dependent manner. At concentration >10(-5) Cd also induced a general toxic effect in S. palustre, leading to chlorophyll degradation and moss death. Moreover, we followed the fate of supplied heavy metals into the moss tissue by SEM-EDX to see if they entered the cells. SEM-EDX observations on moss cultures treated with equimolar concentrations of the two metals showed that most Pb precipitated in form of particles on moss surface, while Cd did not aggregate in particles and was not found on moss surface. In light of these findings, we concluded that probably Pb induced a genotoxic effect at lower intracellular concentrations than Cd. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Analysis of genetic diversity of a native population of Myrcia lundiana Kiaersk. plants using ISSR markers.

    Science.gov (United States)

    Alves, M F; Nizio, D A C; Brito, F A; Sampaio, T S; Silva, A V C; Arrigoni-Blank, M F; Carvalho, S V A; Blank, A F

    2016-12-02

    Myrcia lundiana Kiaersk. is a tree of the family Myrtaceae found in tropical and subtropical areas of the southern hemisphere that produces essential oil. The aim of this study was to characterize the genetic diversity of M. lundiana plants from a native population of Parque Nacional de Itabaiana, using inter-simple sequence repeat molecular markers. Thirty-five primers were tested, 20 of which were polymorphic, resulting in 135 polymorphic and informative bands. Results of the cluster analysis, obtained using the unweighted pair group method with arithmetic mean, grouped plants into three clusters: Cluster I - MLU001, MLU002, MLU003, MLU004, MLU005, MLU006, MLU018, MLU019, MLU020, MLU021, MLU022; MLU008, MLU011, MLU012, MLU014, MLU015, MLU017, MLU026, and MLU028; Cluster II - MLU007, MLU009, MLU010, MLU013, and MLU016; and Cluster III - MLU023, MLU024, MLU025, and MLU027. Jaccard similarity coefficients for pair-wise comparisons of plants ranged between 0.15 and 0.87. MLU014 and MLU015 presented low genetic diversity, with a similarity index of 0.87. Conversely, MLU007 and MLU019 presented high diversity, with a similarity index of 0.15. According to the structure analysis, three distinct clusters were formed. Genetic diversity of M. lundiana plants was intermediate, and expansion of its genetic diversity is necessary. MLU026 and MLU028 are the most suitable for selection in breeding programs, since they clearly represent all of the diversity present in these plants. Moreover, these results provide important information on the existing genetic variability, highlighting the importance of Parque Nacional de Itabaiana for the conservation of this species.

  3. Evaluation of genetic diversity of cultivated tea clones (Camellia sinensis (L. Kuntze in the eastern black sea coast by inter-simple sequence repeats (ISSRS

    Directory of Open Access Journals (Sweden)

    Beris Fatih S.

    2016-01-01

    Full Text Available Tea is the most globally consumed drink after spring water and an important breeding plant with high economical value in Turkey. In half a century, various kinds of tea cultivars have been bred in Turkey to improve the quality and yield of tea plants. Since tea reproduces sexually, tea fields vary in quality. Thus, determining the genetic diversity and relationship of the plants to support breeding and cultivation is important. In this study we aimed to determine the genetic diversity of tea cultivars breeding in the Eastern Black Sea coast of Turkey and the genetic relationship between them, to verify whether the qualitative morphological designations of the clones are genetically true by the ISSR markers. Herein, the genetic diversity and relationships of 18 Turkish tea cultivars were determined using 15 ISSR markers with sizes ranging from 250 to 3000 base pairs. The similarity indices among these cultivars were between 0.456 and 0.743. Based on cluster analysis using UPGMA, some of tea cultivars originating from the same geographical position were found to be clustered closely. Our data provide valuable information and a useful basis to assist selection and cloning experiments of tea cultivars and also help farmers to find elite parental clones for tea breeding in the Eastern Black Sea coast of Turkey.

  4. Automated genotyping of dinucleotide repeat markers

    Energy Technology Data Exchange (ETDEWEB)

    Perlin, M.W.; Hoffman, E.P. [Carnegie Mellon Univ., Pittsburgh, PA (United States)]|[Univ. of Pittsburgh, PA (United States)

    1994-09-01

    The dinucleotide repeats (i.e., microsatellites) such as CA-repeats are a highly polymorphic, highly abundant class of PCR-amplifiable markers that have greatly streamlined genetic mapping experimentation. It is expected that over 30,000 such markers (including tri- and tetranucleotide repeats) will be characterized for routine use in the next few years. Since only size determination, and not sequencing, is required to determine alleles, in principle, dinucleotide repeat genotyping is easily performed on electrophoretic gels, and can be automated using DNA sequencers. Unfortunately, PCR stuttering with these markers generates not one band for each allele, but a pattern of bands. Since closely spaced alleles must be disambiguated by human scoring, this poses a key obstacle to full automation. We have developed methods that overcome this obstacle. Our model is that the observed data is generated by arithmetic superposition (i.e., convolution) of multiple allele patterns. By quantitatively measuring the size of each component band, and exploiting the unique stutter pattern associated with each marker, closely spaced alleles can be deconvolved; this unambiguously reconstructs the {open_quotes}true{close_quotes} allele bands, with stutter artifact removed. We used this approach in a system for automated diagnosis of (X-linked) Duchenne muscular dystrophy; four multiplexed CA-repeats within the dystrophin gene were assayed on a DNA sequencer. Our method accurately detected small variations in gel migration that shifted the allele size estimate. In 167 nonmutated alleles, 89% (149/167) showed no size variation, 9% (15/167) showed 1 bp variation, and 2% (3/167) showed 2 bp variation. We are currently developing a library of dinucleotide repeat patterns; together with our deconvolution methods, this library will enable fully automated genotyping of dinucleotide repeats from sizing data.

  5. 基于ISSR标记的稻属AA基因组遗传多样性分析%ANALYSIS ON GENETIC DIVERSITY OF AA-GENOME ORYZA SPECIES BY ISSR MARKERS

    Institute of Scientific and Technical Information of China (English)

    段世华; 郑卓; 罗强; 龙伟雄; 廖佛才

    2013-01-01

      In order to determine genetic diversity of the AA-genome Oryza species (Poaceae), inter-simple sequence repeat (ISSR) markers from a total of 62 rice accessions collected worldwide were analyzed. These accessions encompassed six wild (O. nivara, O. rufipogon, O. barthii, O. longistaminata, O. glumaepatula, and O. meridionalis) and two cultivated (O. sativa and O. glaberrima) species. 21 selected ISSR primers that produced consistent and repeatable banding patterns revealed significant polymorphisms among the 62 rice accessions with an overall gene diversity (DG) of 0.527, indicating the power of ISSR markers in studying genetic diversity in Oryza germplasm. The consensus tree constructed on the basis of the pairwise Jaccard similarity coefficients of the ISSR banding pattern revealed an evident genetic variation relationships of the AA-genome Oryza species with high bootstrap value supports. It is concluded from this study that the Oryza species from different continents possessed close linkages and current classification of the AA-genome Oryza species suggested by Vaughan (1989) remains valid, particularly in relation to that of the Asian wild rice. The knowledge will be useful for the effective utilization of AA-genome wild Oryza species in rice breeding programs.%  为了确定稻属AA基因组物种间的遗传差异和系统进化关系,62份来自广泛地理分布的水稻品系被用于 ISSR 标记分析。这些品系包含有6个野生稻种(O. nivara, O. rufipogon、O. barthi, O. longistaminata, O. glumaepatula,和O. meridionalis)和2个栽培稻种(O. sativa 和O. glaberrima)。21条能产生良好重复性条带模式的ISSR引物被筛选出,并在62个水稻品系中揭示出非常好的多态性。全部样品的基因多样性为0.527,同时显示出ISSR标记在稻属物种遗传多样性研究中具有强大的作用。根据ISSR条带模式,利用Jaccard配对相似系数构建的一致性树状图,显示出具有良好

  6. SSR和ISSR分子标记及其在桑树遗传育种研究中的应用前景%SSR and ISSR Molecular Markers and Their Usage in Genetics and Breeding of Mulberry Tress

    Institute of Scientific and Technical Information of China (English)

    赵卫国; 苗雪霞; 潘一乐; 黄勇平

    2006-01-01

    本文对SSR(simple sequence repeat)和ISSR(inter-simple sequence repeat)分子标记的定义和各自优点进行了综述,并展望这两种分子标记技术在桑树遗传育种研究中的应用前景.

  7. ISSR Marker Based Population Genetic Study of Melocanna baccifera (Roxb. Kurz: A Commercially Important Bamboo of Manipur, North-East India

    Directory of Open Access Journals (Sweden)

    Heikrujam Nilkanta

    2017-01-01

    Full Text Available Melocanna baccifera (Roxb. Kurz is an economically important bamboo of North-East India experiencing population depletion in its natural habitats. Genetic variation studies were conducted in 7 populations sampled from 5 districts of Manipur using ISSR molecular markers. The investigation was carried out as a primary step towards developing effective conservation strategies for the protection of bamboo germplasm. ISSR marker analysis showed significant level of genetic variation within the populations as revealed by moderately high average values of Nei’s genetic diversity (H 0.1639, Shannon’s diversity index (I 0.2563, percentage of polymorphic bands (PPB 59.18, total genetic variation (Ht 0.1961, and genetic diversity within population (Hs 0.1639. The study also divulged a high genetic variation at species level with Shannon’s diversity index (I, Nei’s genetic diversity (H, and percentage of polymorphic band (PPB% recorded at 0.3218, 0.1939, and 88.37, respectively. Genetic differentiation among the populations (Gst was merely 19.42% leaving 80.58% of genetic variation exhibited within the populations. The low genetic diversity between populations was consistent with AMOVA. The low genetic differentiation among populations coupled with existence of significantly high genetic diversity at species level indicated the urgent necessity of preserving and protecting all the existing natural bamboo populations in the region.

  8. ISSR Marker Based Population Genetic Study of Melocanna baccifera (Roxb.) Kurz: A Commercially Important Bamboo of Manipur, North-East India.

    Science.gov (United States)

    Nilkanta, Heikrujam; Amom, Thoungamba; Tikendra, Leimapokpam; Rahaman, Hamidur; Nongdam, Potshangbam

    2017-01-01

    Melocanna baccifera (Roxb.) Kurz is an economically important bamboo of North-East India experiencing population depletion in its natural habitats. Genetic variation studies were conducted in 7 populations sampled from 5 districts of Manipur using ISSR molecular markers. The investigation was carried out as a primary step towards developing effective conservation strategies for the protection of bamboo germplasm. ISSR marker analysis showed significant level of genetic variation within the populations as revealed by moderately high average values of Nei's genetic diversity (H 0.1639), Shannon's diversity index (I 0.2563), percentage of polymorphic bands (PPB 59.18), total genetic variation (Ht 0.1961), and genetic diversity within population (Hs 0.1639). The study also divulged a high genetic variation at species level with Shannon's diversity index (I), Nei's genetic diversity (H), and percentage of polymorphic band (PPB%) recorded at 0.3218, 0.1939, and 88.37, respectively. Genetic differentiation among the populations (Gst) was merely 19.42% leaving 80.58% of genetic variation exhibited within the populations. The low genetic diversity between populations was consistent with AMOVA. The low genetic differentiation among populations coupled with existence of significantly high genetic diversity at species level indicated the urgent necessity of preserving and protecting all the existing natural bamboo populations in the region.

  9. ISSR Marker Based Population Genetic Study of Melocanna baccifera (Roxb.) Kurz: A Commercially Important Bamboo of Manipur, North-East India

    Science.gov (United States)

    Nilkanta, Heikrujam; Amom, Thoungamba; Rahaman, Hamidur

    2017-01-01

    Melocanna baccifera (Roxb.) Kurz is an economically important bamboo of North-East India experiencing population depletion in its natural habitats. Genetic variation studies were conducted in 7 populations sampled from 5 districts of Manipur using ISSR molecular markers. The investigation was carried out as a primary step towards developing effective conservation strategies for the protection of bamboo germplasm. ISSR marker analysis showed significant level of genetic variation within the populations as revealed by moderately high average values of Nei's genetic diversity (H 0.1639), Shannon's diversity index (I 0.2563), percentage of polymorphic bands (PPB 59.18), total genetic variation (Ht 0.1961), and genetic diversity within population (Hs 0.1639). The study also divulged a high genetic variation at species level with Shannon's diversity index (I), Nei's genetic diversity (H), and percentage of polymorphic band (PPB%) recorded at 0.3218, 0.1939, and 88.37, respectively. Genetic differentiation among the populations (Gst) was merely 19.42% leaving 80.58% of genetic variation exhibited within the populations. The low genetic diversity between populations was consistent with AMOVA. The low genetic differentiation among populations coupled with existence of significantly high genetic diversity at species level indicated the urgent necessity of preserving and protecting all the existing natural bamboo populations in the region. PMID:28168084

  10. Genetic diversity of Pleurotus pulmonarius revealed by RAPD, ISSR, and SRAP fingerprinting.

    Science.gov (United States)

    Yin, Yonggang; Liu, Yu; Li, Huamin; Zhao, Shuang; Wang, Shouxian; Liu, Ying; Wu, Di; Xu, Feng

    2014-03-01

    Pleurotus pulmonarius is one of the most widely cultivated and popular edible fungi in the genus Pleurotus. Three molecular markers were used to analyze the genetic diversity of 15 Chinese P. pulmonarius cultivars. In total, 21 random amplified polymorphic DNA (RAPD), 20 inter-simple sequence repeat (ISSR), and 20 sequence-related amplified polymorphism (SRAP) primers or primer pairs were selected for generating data based on their clear banding profiles produced. With the use of these RAPD, ISSR, and SRAP primers or primer pairs, a total of 361 RAPD, 283 ISSR, and 131 SRAP fragments were detected, of which 287 (79.5 %) RAPD, 211 (74.6 %) ISSR, and 98 (74.8 %) SRAP fragments were polymorphic. Unweighted Pair-Group Method with Arithmetic Mean (UPGMA) trees of these three methods were structured similarly, grouping the 15 tested strains into four clades. Subsequently, visual DNA fingerprinting and cluster analysis were performed to evaluate the resolving power of the combined RAPD, ISSR, and SRAP markers in the differentiation among these strains. The results of this study demonstrated that each method above could efficiently differentiate P. pulmonarius cultivars and could thus be considered an efficient tool for surveying genetic diversity of P. pulmonarius.

  11. Verification of the systematic position of California brome (Bromus carinatus Hook. and Arn., Poaceae, cv. 'Broma', on the basis of analysis of issr markers

    Directory of Open Access Journals (Sweden)

    Agnieszka Sutkowska

    2012-10-01

    Full Text Available ‘Broma’ is a grass cultivar belonging to the species Bromus carinatus. In the Lists of Agricultural Plant Varieties of the Research Centre for Cultivar Testing (COBORU, it is shown as Bromus willdenowii (= B. catharticus, B. unioloides (List of Agricultural Plant Varieties 1989-2009, whereas already in 1984 Mirek demonstrated on the basis of morphological analysis that this was a different closely related species – B. carinatus. The aim of the present study was to verify the species affiliation of cv. ‘Broma’. The conducted analysis of ISSR molecular markers included representatives of cv. ‘Broma” as well as of B. carinatus and B. willdenowii. The method used allowed the identification of molecular markers of the above-mentioned taxa. The numerical analysis of the obtained results suggests that cv. ‘Broma’ should be classified in the species B. carinatus, not B. willdenowii.

  12. 广西壳菜果遗传多样性的ISSR研究%Genetic Diversity of the Mytilaria laosensis in Guangxi Detected by ISSR Markers

    Institute of Scientific and Technical Information of China (English)

    彭继庆; 曹福祥; 许若娴

    2012-01-01

    利用ISSR分子标记技术,对广西壳菜果4个自然种群共43个个体进行了遗传多样性研究.结果表明,广西壳菜果天然种群总体水平的遗传多样性较高,各个种群的遗传多样性水平存在差异,其中广西凭祥种群的遗传多样性水平最高,广西那坡种群的最低.通过PopGen32分析表明:有82.43%的遗传变异存在于种群内,而17.57%的遗传变异存在于种群间,遗传分化系数(Gst)为0.175 8,种群间的基因流(Nm)为2.344 7,种群间的遗传分化程度较低,基因流动足以抵制遗传漂变的影响.利用UPGMA法对4个种群进行聚类分析显示:广西凭祥、广西德保和广西靖西3个种群聚为一类,广西那坡种群单独聚为一类.%The genetic diversity of 43 individuals of Mytilaria laosensis populations in Guangxi province was studied using inter-simple sequence repeat(ISSR) markers. The results show that the general level of Mytilaria laosensis natural population genetic diversity in Guangxi is high. The genetic diversity level of four populations is different largely. The level of genetic diversity of Pingxiang population in Guangxi is the highest among the four populations , and that of Napo population in Guangxi is the lowest. The analysis of genetic differentiation shows that the 82.43% of genetic variation is in population, and the others is not in population by PopGen 32. Coefficient of genetic differentiation (Gst) among four populations is 0. 175 8 and the gene flow (Nm) among Mytilaria laosensis is 2. 344 7. It shows that the level of genetic differentiation is low and gene flow is sufficient to resist the effects of genetic drift. UPGMA cluster analysis shows that four populations can be classified two kinds, and the population of Pingxiang, Debao and Jingxi in Guangxi are divided into one kind and the population of Napo in Guangxi is divided into one kind alone.

  13. Inter simple sequence repeat fingerprints for assess genetic diversity of tunisian garlic populations

    OpenAIRE

    Jabbes, Naouel; Geoffriau, Emmanuel; Le Clerc, Valérie; Dridi, Boutheina; Hannechi, Chérif

    2011-01-01

    Garlic (Allium sativum L.) that is cultivated in Tunisia is heterogeneous and unclassified with no registered local cultivars. At present, the level of genetic diversity in Tunisian garlic is almost unknown. Inter Simple Sequence Repeats (ISSR) genetic markers were therefore used to assess the genetic diversity and its distribution in 31 Tunisian garlic accessions with 4 French classified clones used as control. It was the first time that ISSR markers were used to detect diversity in garlic. ...

  14. 24份葡萄种质亲缘关系的ISSR分析%Analysis of genetic relationship of 24 Vitis germplasm resources by ISSR markers

    Institute of Scientific and Technical Information of China (English)

    李琳; 魏灵珠; 程建徽; 梅军霞; 吴江

    2013-01-01

    利用ISSR标记对24份葡萄材料进行了基因组多态性分析,从50条引物中筛选出6条扩增稳定且多态性丰富的引物用于葡萄的ISSR扩增.共扩增出59条条带,其中多态性条带53条,多态性百分率为90%.根据ISSR扩增结果,利用NTSYSpc2.10e软件进行Jaccard相似性系数分析,24份葡萄材料的遗传相似系数为0.42 ~0.88,平均遗传相似系数为0.6495.在遗传相似系数为0.55处,24份葡萄材料明显分为2大类群.第1类包含10个欧美杂种、7个欧亚种、1个华欧杂种,第2类包含3个美洲杂交种、1个河岸葡萄、1个冬葡萄、1个东亚葡萄.由此可见,美洲杂交种与欧美杂种、欧亚种葡萄的亲缘关系较远,欧美杂种与欧亚种葡萄之间亲缘关系较近.此外,引物BC820与引物BC847分别在我国自主选育砧木品种华佳8号与抗砧3号中扩增出一条特异条带,可为利用ISSR标记鉴定葡萄品种或品系提供参考依据.%Twenty-four Vitis germplasm were used as materials for analyzing their genome polymorphism by ISSR markers.Six primers with stable amplification and rich polymorphism selected from 50 primers and used for ISSR amplification.A total of 59 bands were generated,of which 53 bands were polymorphic bands (90.0%).Based on ISSR amplification and analysis by NTSYSpc2.10e software,the genetic similarity coefficient varied from 0.42 to 0.88 with an average of 0.6495.The clustering dendrogram was constructed by UPGMA method.Twenty-four Vitis materials were divided into two major groups at the similarity of 0.55.The first group included 10 cultivars of Euro-American hybrids,7 cultivars of V.vinifera and 1 cultivar of China European hybrid(V.pseudoreticulat × V.vinifera).The second group included 3 culiivars of American hybrids,1 cultivar of Riverside grape,1 cultivar of V.berlandieri grape and 1 cultivar of East Asia grape.The results showed that American hybrids and Euro-American hybrids had a distant relationship

  15. Genetic diversity analyses of Lasiodiplodia theobromae on Morus alba and Agave sisalana based on RAPD and ISSR molecular markers

    Directory of Open Access Journals (Sweden)

    Hong-hui Xie

    2016-10-01

    Full Text Available Genetic diversity of 23 Lasiodiplodia theobromae isolates on Morus alba and 6 isolates on Agave sisalana in Guangxi province, China, was studied by using random amplified polymorphic DNA and inter-simple sequence repeat molecular markers. Results of two molecular markers showed that the average percentage of polymorphic loci of all isolates was more than 93%. Both dendrograms of two molecular markers showed obvious relationship between groups and the geographical locations where those strains were collected, among which, the 23 isolates on M. alba were divided into 4 populations and the 6 isolates on A. sisalana were separated as a independent population. The average genetic identity and genetic distance of 5 populations were 0.7215, 0.3284 and 0.7915, 0.2347, respectively, which indicated that the genetic identity was high and the genetic distance was short in the 5 populations. Average value of the gene diversity index (H and the Shannon’s information index (I of 29 isolates were significantly higher than 5 populations which showed that genetic diversity of those isolates was richer than the populations and the degree of genetic differentiation of the isolates was higher. The Gst and Nm of 29 isolates were 0.4411, 0.6335 and 0.4756, 0.5513, respectively, which showed that the genetic diversity was rich in those isolates.

  16. Analysis of drought-tolerant sugar beet (Beta vulgaris L.) mutants induced with gamma radiation using SDS-PAGE and ISSR markers.

    Science.gov (United States)

    Sen, Ayse; Alikamanoglu, Sema

    2012-01-01

    Drought is one of the major environmental stresses which greatly affect the plant growth and productivity. In the present study, various doses (0-75Gy) of gamma rays were applied to investigate the effect of radiation on shoot tip explants. It was observed that the regeneration rates and plant fresh weights decreased significantly with an increase in radiation dose. The optimal irradiation doses for mutation induction were determined at 15 and 20Gy. Afterwards, the induction of somatic mutation in sugar beet (Beta vulgaris L.) was investigated by irradiation of shoot tips with 15 and 20Gy gamma rays. Irradiated shoot tips were sub-cultured and M(1)V(1)-M(1)V(3) generations were obtained. Mutants tolerant to drought stress were selected on MS medium, supplemented with 10 and 20gl(-1) PEG6000. Of the M(1)V(3) plantlets, drought-tolerant mutants were selected. Leaf soluble proteins obtained from the control and drought-tolerant mutants were analyzed by SDS-PAGE. A total of 22 protein bands were determined and 2 of them were observed to be drought-tolerant mutants except the control. Polymorphism was also detected among the control and drought-tolerant mutants by DNA fingerprinting using ISSR markers. A total of 106 PCR fragments were amplified with 19 ISSR primers and 91 of them were polymorphic. The dendrograms were separated into two main clusters. First cluster included M8 mutant plant, which was applied 20Gy gamma radiation and regenerated on selective culture media containing 10gl(-1) PEG6000 concentration, and the second cluster was further divided into five sub-clusters.

  17. Analysis of drought-tolerant sugar beet (Beta vulgaris L.) mutants induced with gamma radiation using SDS-PAGE and ISSR markers

    Energy Technology Data Exchange (ETDEWEB)

    Sen, Ayse, E-mail: senayse@istanbul.edu.tr [Istanbul University, Faculty of Science, Department of Biology, 34459 Vezneciler, Istanbul (Turkey); Alikamanoglu, Sema [Istanbul University, Faculty of Science, Department of Biology, 34459 Vezneciler, Istanbul (Turkey)

    2012-10-15

    Drought is one of the major environmental stresses which greatly affect the plant growth and productivity. In the present study, various doses (0-75 Gy) of gamma rays were applied to investigate the effect of radiation on shoot tip explants. It was observed that the regeneration rates and plant fresh weights decreased significantly with an increase in radiation dose. The optimal irradiation doses for mutation induction were determined at 15 and 20 Gy. Afterwards, the induction of somatic mutation in sugar beet (Beta vulgaris L.) was investigated by irradiation of shoot tips with 15 and 20 Gy gamma rays. Irradiated shoot tips were sub-cultured and M{sub 1}V{sub 1}-M{sub 1}V{sub 3} generations were obtained. Mutants tolerant to drought stress were selected on MS medium, supplemented with 10 and 20 gl{sup -1} PEG6000. Of the M{sub 1}V{sub 3} plantlets, drought-tolerant mutants were selected. Leaf soluble proteins obtained from the control and drought-tolerant mutants were analyzed by SDS-PAGE. A total of 22 protein bands were determined and 2 of them were observed to be drought-tolerant mutants except the control. Polymorphism was also detected among the control and drought-tolerant mutants by DNA fingerprinting using ISSR markers. A total of 106 PCR fragments were amplified with 19 ISSR primers and 91 of them were polymorphic. The dendrograms were separated into two main clusters. First cluster included M8 mutant plant, which was applied 20 Gy gamma radiation and regenerated on selective culture media containing 10 g l{sup -1} PEG6000 concentration, and the second cluster was further divided into five sub-clusters.

  18. Genetic Diversity of Selected Mangifera Species Revealed by Inter Simple Sequence Repeats Markers

    OpenAIRE

    Zulhairil Ariffin; Muhammad Shafie Md Sah; Salma Idris; Nuradni Hashim

    2015-01-01

    ISSR markers were employed to reveal genetic diversity and genetic relatedness among 28 Mangifera accessions collected from Yan (Kedah), Bukit Gantang (Perak), Sibuti (Sarawak), and Papar (Sabah). A total of 198 markers were generated using nine anchored primers and one nonanchored primer. Genetic variation among the 28 accessions of Mangifera species including wild relatives, landraces, and clonal varieties is high, with an average degree of polymorphism of 98% and mean Shannon index, H0=7.5...

  19. ISSR分子标记及其在中国南方果树上的应用%Application of ISSR molecular markers on southern fruit trees in China

    Institute of Scientific and Technical Information of China (English)

    宋晓兵; 彭埃天; 刘景梅; 陈霞

    2009-01-01

    ISSR is a new type of molecular marker technology, based the sequence of microsatellite, and it is simple, stable,DNA polymorphism high. In this paper, the principle of ISSR marker technology, characteristics, and germplasm identification,genetic relationship analysis, genetic diversity of the fruit trees in southern China, as well as such areas as genetic mapping research and application are reviewed.%ISSR是一种基于微卫星序列发展起来的新型分子标记技术,具有简便、稳定、DNA多态性高等优点.从ISSR标记技术的原理、特点及其在中国南方果树种质鉴定、亲缘关系分析、遗传多样性、以及遗传作图等方面的研究应用进行了综述.

  20. 卷柏科药用植物种间遗传多样性的ISSR分析%Genetic diversity analysis of medicinal plants of Selaginellaceae based on ISSR markers

    Institute of Scientific and Technical Information of China (English)

    孙红梅; 谷巍; 耿超; 孙庆文; 曹园

    2016-01-01

    应用ISSR分子标记技术对34种卷柏科植物(其中18种药用植物)进行遗传多样性分析。利用PopGene 32软件及NTsys 2.10e软件分析卷柏科34种植物的遗传多样性及亲缘关系,并根据UPGMA法,构建亲缘关系树状图。从100个随机引物中筛选出7条多态性稳定、条带清晰的引物,共扩增出156条谱带,多态性比例为100%。分析结果表明,平均Shannon信息指数为0.537,平均 Nei’s基因多样性指数为0.359,平均有效等位基因数为1.617,34种卷柏科植物种间的遗传相似性系数在0.359~0.872,显示出该植物类群具丰富的遗传多样性。聚类分析结果表明,34种卷柏科植物之间有较明显的分界,其中卷柏与垫状卷柏,剑叶卷柏与异穗卷柏,兖州卷柏和江南卷之间的亲缘关系较近,分别聚为一类。初步证明利用ISSR分子标记可有效地分析卷柏科药用植物的遗传多样性,从而为卷柏科植物的分类鉴定以及资源的合理开发利用提供理论基础和科学依据。%ISSR (inter simple sequence repeat) markers was used to analyze the genetic diversity and related-ness of 34 species of Selaginellaceae (18 medicinal species included). The molecular marker technique ISSR was used to investigate the genetic diversity and relatedness of 34 species of Selaginellaceae. Data was ana-lyzed by PopGene 32 and NTsys 2.10e, and a cluster diagram was presented by UPGMA. 100 primers were tested, among which seven gave reliable polymorphic banding patterns. Ampliifcation from the accessions of the seven primers yielded 156 genetic loci with 100% polymorphism. The results from PopGene 32 software indicated that the average Shannon information index was 0.537, the average Nei’s gene diversity index was 0.359, the average effective number of alleles was 1.617, and the genetic similarity coefifcient of varieties was between 0.359 and 0.872, which showed enriched genetic diversity. There are obvious boundaries between 34 species

  1. SSR marker, ISSR marker and their application to plant genetics and breeding%SSR和ISSR分子标记及其在植物遗传育种研究中的应用

    Institute of Scientific and Technical Information of China (English)

    张立荣; 徐大庆; 刘大群

    2002-01-01

    SSR(Simple Sequence Repeat)和ISSR(Inter-Simple Sequence Repeat)技术是在PCR基础上发展起来的两种DNA多态性检测技术,已开始应用于基因组研究的各个领域.概述了SSR、ISSR反应的原理、特点,总结了其在植物亲缘关系和遗传多态性研究、DNA指纹库的建立、遗传图谱的构建和基因定位及分子标记辅助育种等方面的应用,并肯定了SSR、ISSR在植物遗传育种领域的广阔应用前景.

  2. Relationship Between Hybrid Performance and Genetic Diversity Based on SSRs and ISSRs in Brassica napus L.

    Institute of Scientific and Technical Information of China (English)

    SHEN Jin-xiong; FU Ting-dong; YANG Guang-sheng

    2003-01-01

    To investigate the relationship between genetic distance (GD) and hybrid performance, twotypes of molecular markers, microsatellites (simple sequence repeats, SSRs) and intro-simple sequence repeats(ISSRs), were employed to detect the genetic diversity of 3 double low self-incompatible lines and 22 male pa-rental varieties of Brassica napus from different geographical origins. Hybrids were produced in a NC Ⅱ mat-ing design by hand-pollination. The result indicated that 25 parental varieties (lines) could be divided into sixgroups by Un-weighted Pair Group Mathematics Average (UPGMA) clustering based on GDs. SI-1300 and SI-1320 could be singly clustered into one group, respectively. Varieties from China could be separated into an-other group, SI-1310 and varieties from foreign countries could be separated into other three groups. Thegrouping was generally consistent with parental pedigrees and geographical origins. Significant differences inyield, quality and phenological period traits were observed among these parent groups. Although hybrid yield/plant showed significantly positive correlation with genetic distance based on SSR and ISSR markers, but thedetermination coefficient was iow. It appeared to be unsuitable for using the genetic distance based on SSR andISSR markers to predict heterosis and hybrid performance in Brassica napus.

  3. Highly Informative Simple Sequence Repeat (SSR) Markers for Fingerprinting Hazelnut

    Science.gov (United States)

    Simple sequence repeat (SSR) or microsatellite markers have many applications in breeding and genetic studies of plants, including fingerprinting of cultivars and investigations of genetic diversity, and therefore provide information for better management of germplasm collections. They are repeatab...

  4. simple sequence repeat (SSR) markers in genetic analysis of

    African Journals Online (AJOL)

    Yomi

    2012-08-28

    Aug 28, 2012 ... In the present study, 78 mapped simple sequence repeat (SSR) markers representing 11 ... mean (UPGMA) with each cluster representing a particular Vigna species. ..... were reported to be more frequent than the compound.

  5. Evolutionary analysis of allotetraploid hybrids of red crucian carp × common carp,based on ISSR,AFLP molecular markers and cloning of cyclins genes

    Institute of Scientific and Technical Information of China (English)

    LIU LiangGuo; YAN JinPeng; LIU ShaoJun; LIU Dong; YOU CuiPing; ZHONG Huan; TAO Min; LIU Yun

    2009-01-01

    The allotetraploid hybrids of red crucian carp × common carp are the first reported artificially cultured polyploid fish with bisexual fertility and stable inheritance in vertebrate.Using ISSR and AFLP markers and the cyclins genes,the genomes and cyclin gene sequence changes were analyzed between the allotetraploid hybrids and their parents.The results indicated that the allotetraploids inherited many genetic characteristics from their parents and the genetic characteristics were stable after 15 generations.However,the allotetraploids had a closer genetic relationship with their original female parents and represented a bias toward the maternal progenitor.DNA fingerprinting analysis showed that the allotetraploids had undergone sequences deletion from their original parents and that the deleted sequences were mostly from the male parent's genome.Some non-parental bands were found in the allotetraploid hybrids.Sequences analysis of the cyclin A1 and B1 genes showed nonsynonymous substitutions of single nucleotides in codons that were different from their original parents,leading to non-parental amino acid loci.We speculate that the non-additivity in the allotetraploids,compared with their progenitors,could be an adjustment to the genomic shock from heterozygosity and polyploidy, allowing maintenance of genetic stability.

  6. COMPARATIVE ANALYSIS OF INTER SIMPLE SEQUENCE REPEATS AND SIMPLE SEQUENCE REPEATS MARKERS: GENETIC ANALYSIS OF DESCHAMPSIA CESPITOSA POPULATIONS GROWING IN METAL CONTAMINATED REGIONS IN CANADA

    Directory of Open Access Journals (Sweden)

    K.K. Nkongolo

    2014-01-01

    Full Text Available Comparative studies conducted on the genetic variation of metal-tolerant populations and their non-metal-tolerant counterparts have been performed on numerous species using isozyme markers. Analysis of genetic differences among plant populations growing in heavy metal-contaminated and uncontaminated regions are limited. The main objectives of the present study were to compare ISSR and microsatellite markers in assessing genetic variation in D. cespitosa populations that colonized metal-contaminated and uncontaminated regions in Northern Ontario, Canada. Total genomic DNA from D. cespitosa samples were amplified with ISSR and SSR primers using optimized PCR conditions. The level of polymorphic loci varies from 46 to 74% for ISSR analysis. The level of observed heterozygosity was moderate to high ranging from 0.44 to 0.68 for the SSR primers used. But no significant difference in genetic variation levels was detected between metal contaminated and uncontaminated sites with SSR markers. There was a significant reduction of polymorphic loci in samples from highly metal-contaminated areas of the Cobalt region compared to the reference sites based on ISSR analysis. Use of a combination of different marker systems is recommended to analyse genetic variation in plant populations.

  7. 辣木植物ISSR分子标记的初步研究%Study on Moringa Plants by ISSR Marker

    Institute of Scientific and Technical Information of China (English)

    韩闯; 李敏; 钟然; 云叶; 杨盛昌

    2008-01-01

    对从7个不同地方采集的7份辣木(Moringa)材料进行了ISSR 标记分析.结果表明,被测材料间ISSR标记多态性较高.从18个ISSR 引物中筛选出6个可扩增出清晰的且具多态性的条带的引物,共扩增出75条带,其中59条具有多态性, 占总扩增带数的78.67 %,其中每个引物可扩增出4~14条多态性带,平均10.17条.ISSR 标记遗传相似性系数(GS) 变异范围为0.557 0~0.836 7,平均值为0.687 6±0.080 9.聚类分析表明,7份辣木材料可以聚为四类,其中MS2为单独一类.ISSR 标记可以有效地评价辣木植物遗传分化并为辣木种类的鉴别提供一定的理论依据.

  8. 南瓜种质资源遗传多态性的ISSR分析%Analysis of Genetic Diversity in Squash by ISSR Markers

    Institute of Scientific and Technical Information of China (English)

    朱海生; 卢丽芳; 陈敏氡; 林珲; 王彬; 温庆放; 林义章

    2015-01-01

    本研究应用ISSR分子标记技术对85份南瓜种质资源进行遗传多样性分析。结果显示:从100条ISSR引物中筛选出12条引物进行PCR扩增,共获得条带125条,多态性占115条,10条条带为所有南瓜品种所共有,平均每个引物的条带数为10.42条,多态性比例为92%。通过Ntsys2.10e软件计算出85份南瓜资源的遗传相似系数0.3840~1.0000之间。通过聚类分析将所收集的南瓜品种分为三类:分别是美洲南瓜、中国南瓜和印度南瓜,其中亲缘关系较近的是中国南瓜和美洲南瓜,中国南瓜和美洲南瓜与印度南瓜的亲缘关系均比较远。本研究为南瓜种质资源的保护、利用及新品种的培育提供了科学依据。%In this study, the genetic diversity of 85 accessions in Squash was revealed by ISSR markers. 12 polymorphic primers were screened out of 100 polymorphic primers, which generated 125 DNA bands with average 10.42 bands each primer. 115 bands were polymorphic, 10 bands were shared with all tested primers. The ratio of polymorphic bands was 92%, and the genetic similarity coefficient of 85 accessions in squash ranged between 0.384 0 to 1.000 0, which was calculated by Ntsys2.10e. The cluster analysis showed that squashes could be divided into three groups: Cucurbita pepo L., Cucurbita moschata, and Cucurbita maxima. The relationship of Cucurbita moschata and Cucurbita pepo L. was close, whereas their kinship with the Cucurbita maxima was far. This study provided scientific basis for the protection and utilization of squash germplasm resources and cultivation of new varieties.

  9. High gene flow and genetic diversity in three economically important Zanthoxylum Spp. of Upper Brahmaputra Valley Zone of NE India using molecular markers

    Directory of Open Access Journals (Sweden)

    K. Medhi

    2014-12-01

    Full Text Available The genetic diversity in Zanthoxylum species viz. Zanthoxylum nitidum, Zanthoxylum oxyphyllum and Zanthoxylum rhesta collected from the Upper Brahmaputra Valley Zone of Assam (NE India was amplified using 13 random amplified polymorphic DNA (RAPD markers and 9 inter-simple sequence repeat (ISSR markers. RAPD markers were able to detect 81.82% polymorphism whereas ISSR detected 98.02% polymorphism. The genetic similarities were analyzed from the dendrogram constructed by RAPD and ISSR fingerprinting methods which divided the 3 species of Zanthoxylum into 3 clear different clusters. The principle component analysis (PCA was carried out to confirm the clustering pattern of RAPD and ISSR analysis. Analysis of molecular variance (AMOVA revealed the presence of significant variability between different Zanthoxylum species and within the species by both RAPD and ISSR markers. Z. nitidum was found to be sharing a high degree of variation with the other two Zanthoxylum species under study. The Nei's gene diversity (h, Shannon's information index (I, observed number of alleles (na and effective number of alleles (ne were also found to be higher in ISSR markers (0.3526, 0.5230, 1.9802 and 1.6145 than in RAPD markers (0.3144, 0.4610, 1.8182 and 1.5571. The values for total genotype diversity for among population (HT, within population diversity (Hs and gene flow (Nm were more in ISSR (0.3491, 0.2644 and 1.5610 than RAPD (0.3128, 0.2264 and 1.3087 but the mean coefficient of gene differentiation (GST was more in RAPD (0.2764 than ISSR (0.2426. A comparison of this two finger printing methods was done by calculating MR, EMI and MI. The correlation coefficient between data matrices of RAPD and ISSR based on Mantel test was found to be significant (r = 0.65612.

  10. Phylogenetic study and molecular identification of 31 Dendrobium species using inter-simple sequence repeat (ISSR) markers

    National Research Council Canada - National Science Library

    Wang, H.-Z; Feng, S.-G; Lu, J.-J; Shi, N.-N; Liu, J.-J

    2009-01-01

    The genetic diversity of the genus Dendrobium is not well known and the phylogenetic relationship of Dendrobium species are mainly determined by studies of the comparative vegetative anatomy and plant systematics...

  11. Identification of Soybean Cytoplasmic Male Sterile Line and Maintainer Line with Mitochondrial ISSR and SCAR Markers%线粒体ISSR与SCAR标记鉴定大豆细胞质雄性不育系与保持系

    Institute of Scientific and Technical Information of China (English)

    张春宝; 李玉秋; 彭宝; 王鹏年; 董英山; 赵丽梅

    2013-01-01

    By extracting the mitochondrial DNA of cytoplasmic male sterile line and its maintainer line, screening with 100 ISSR primers, and it was found that the amplification product of primer ISSR829 had differences in sterile line and its maintainer. Then sequenced the different fragments and found that the sequence was only exist in the maintainer mitochondrial genome, and upstream of mitochondrial gene Atp6 by 2 000 bp. It was suggested that the sequence has a relationship with infertility. Designed primers according to the sequence and detected female parent and its maintainer line of five soybean hybrids, and found that sterile line did not appear specific fragment,but a target fragment corresponding maintainer lines. It was confirmed that the specific fragments was converted to a SCAR marker, which could be used for testing the seed purity of cytoplasmic male sterile line.%通过提取细胞质雄性不育系和同型保持系的线粒体基因组DNA,用100条ISSR引物进行筛选,发现引物ISSR829的扩增片段在不育系和保持系间存在差异.对差异片段进行测序发现,该序列仅存在于保持系线粒体基因组中,且位于基因Atp6上游2000 bp处,推断其可能与不育有关.利用测序获得片段设计引物,用目前已育成的5个大豆杂交种的不育系母本和同型保持系的基因组DNA为模版,进行PCR扩增,发现不育系母本未出现特异片段,而对应保持系出现目的片段,表明该特异片段转化为SCAR标记,可用于不育系的筛选和不育系种子纯度鉴定.

  12. Assessment of Genetic Diversities of Selected Laminaria (Laminariales,Phaeophyta) Gametophytes by Inter-Simple Sequence Repeat Analysis

    Institute of Scientific and Technical Information of China (English)

    Xiu-Liang WANG; Chen-Lin LIU; Xiao-Jie LI; Yi-Zhou CONG; De-Lin DUAN

    2005-01-01

    Inter-simple sequence repeat (ISSR) analysis was used to assess genetic diversity among 10pairs of male and female Laminaria gametophytes. A total of 58 amplification loci was obtained from 10selected ISSR primers, of which 34 revealed polymorphism among the gametophytes. Genetic distances were calculated with the Dice coefficient ranging from 0.006 to 0.223. A dendrogram based on the unweighted pair-group method arithmetic (UPGMA) average showed that most male and female gametophytes of the same species were clustered together and that 10 pairs of gametophytes were divided into four groups. This was generally consistent with the taxonomic categories. The main group consisted of six pairs of gametophytes, which were selected from Laminaria japonica Aresch. by intensive inbreeding through artificial hybridization. One specific marker was cloned, but was not converted successfully into a sequence characterized amplified region (SCAR) marker. Our results demonstrate the feasibility of applying ISSR markers to evaluate Laminaria germplasm diversities.

  13. The genetic diversity of strawberry (Fragaria ananassa Duch. hybrids based on ISSR markers - doi: 10.4025/actasciagron.v35i4.16737

    Directory of Open Access Journals (Sweden)

    Claudinéia Ferreira Nunes

    2013-05-01

    Full Text Available The strawberry is an important agricultural crop in Brazil. However, most of the commercial genotypes currently in cultivation in Brazil were developed in other countries with environmental adaptations often inadequate for the regional conditions. In this work, inter-simple sequence repeat markers were used to determine the genetic variability and the loci segregation profiles of 84 strawberry hybrids obtained from a genetic breeding program at the ‘Empresa de Pesquisa Agropecuária de Minas Gerais.’ The hybrids were produced from crosses involving the following progenitors: ‘Toyonoka’ x ‘Sweet Charlie’, ‘Camino Real’ x ‘Sweet Charlie’, ‘Oso Grande’ x ‘Sweet Charlie’, ‘Oso Grande’ x ‘Toyonoka’, ‘Dover’ x ‘Oso Grande’, and ‘Camino Real’ x ‘Toyonoka’. Fourteen genotypes were randomly sampled for each hybrid combination and evaluated. The results showed that the genetic profiles of the hybrids from each test cross were very diverse, most likely due to the high heterozygosity of the genome of each progenitor involved, which might indicate the presence of adequate genetic diversity among the hybrids to allow for the selection of new cultivars with agronomic traits that are more suitable to environmental conditions in Brazil.

  14. Genetic diversity of Salsola passerina populations in northwestern China based on inter-simple sequence repeat(ISSR)%基于ISSR西北地区珍珠猪毛菜(Salsola passerina)的遗传多样性

    Institute of Scientific and Technical Information of China (English)

    高天鹏; 高海宁; 张勇; 徐世健; 安黎哲

    2009-01-01

    Genetic diversity and differentiation among seven populations of Salsola passerina in north-western China were investigated using ISSR markers.15 ISSR primers gave rise to 196 discernible DNA fragments,of which 153(78.06%)were polymorphic.However,the polymorphism at the population level was relatively low,with an average of 45.60%.The mean expected heterozygosity and Shannon,s infor-mation index were 0.1759 and 0.281 1 at the species level,0.125l and 0.1977 at the population level,respectively.Based on Nei's Gst value,a large proportion of genetic variance(70.96%)resided among individuals within populations;however,only 29.04%genetic variance resided among populations.Pair-wise genetic identity values among populations had a mean of 0.932 0.AMOVA analysis also indicated a similar genetic structure.UPGMA cluster analysis based on Nei's genetic distance divided the popula-tions into three main groups:QHDLH,LZXJS and the rest five populations(NXZN,NMWH,MQHLY,JTDX,and AXTS).The mantel test showed that genetic distance was significantly correlated with verti-cal geographical distribution(P<0.001).A means of preserving genetic variation of this endemic species in those regions is to stop deforestation and to protect its suitable habitats in view of the lower genetic diversity within populations.%运片JSSR分子标记的方法,对分布于中国北方7个居群的137个珍珠猪毛菜个体筛选出了15条引物,产生了196条带,其中153条为多态性条带,总的基因多态率为78.06%,多态性在种群水平上较低,平均多态率为45.60%.平均期望杂合度和Shannon信息指数在种间水平上分别为0.1759和0.2811,在居群水平上为0.125l和0.1977,种群内个体间的遗传分化较大,为70.96%,而种群间遗传分化较小,为29.04%,种群间的遗传一致度平均为0.9320.AMOVA分析得到了相似结果.基于Nei's遗传距离的UPGMA聚类分析将7个居群分为3组:德令哈、兰州和其余5个居群(中宁,乌海,民勤,金塔和

  15. 小麦抗叶锈基因Lr37 ISSR分子标记%ISSR Molecular Markers for the Leaf Rust Resistance Gene Lr37 in Wheat

    Institute of Scientific and Technical Information of China (English)

    张立荣; 徐大庆; 杨文香; 刘大群

    2004-01-01

    利用ISSR(内部简单重复序列)技术对Thatcher及20个以Thatcher为轮回亲本的小麦(Triticum aestivum)抗叶锈病(Pucciniareconcita f.sp.tritici)近等基因系(NILs)进行分析,发现1个与Lr37基因连锁的ISSR标记.经过多次重复发现,在100个ISSR引物(UBC801-UBC900)中有2个引物UBC812和UBC848在小麦抗叶锈基因Lr37近等基因系间表现多态性.当用这2个引物对已知含Lr37基因的3个抗病材料及其它不含Lr37基因的感病材料进行检测时,多态性标记UBC812-1200可以从3个含Lr37基因的抗病材料中检测到1条1 200bp的多态性带,而在其它感病材料中,均未出现.进一步用UBC812和UBC848对128株(Thatcher× Lr37/6*Thatcher)F2分离群体进行分析,发现标记UBC812-1200与Lr37基因共分离,可作为该基因的分子标记.

  16. Review of the Methods for Developing SSR Molecular Markers

    Institute of Scientific and Technical Information of China (English)

    ZHAO Xue; CHANG Wei; HAN Yingpeng; LI Wenbin

    2008-01-01

    Microsatellite marker (or Simple Sequence Repeate,SSR) is a marker technology based on DNA molecular length polymorphism.It is also one of the most commonly used molecular markers.Traditional SSR marker development methods are relatively time-consuming and mostly relying on the known genome sequence information while recently developed methods of SSR marker based on RAPD,ISSR-PCR SSR,the use of hybrid options, sequence tag SSR library access and screening EST-SSR have been widely used.This paper gave an overview of the methods mentioned above for the development of SSR markers.

  17. Genetic Diversity of Selected Mangifera Species Revealed by Inter Simple Sequence Repeats Markers

    Directory of Open Access Journals (Sweden)

    Zulhairil Ariffin

    2015-01-01

    Full Text Available ISSR markers were employed to reveal genetic diversity and genetic relatedness among 28 Mangifera accessions collected from Yan (Kedah, Bukit Gantang (Perak, Sibuti (Sarawak, and Papar (Sabah. A total of 198 markers were generated using nine anchored primers and one nonanchored primer. Genetic variation among the 28 accessions of Mangifera species including wild relatives, landraces, and clonal varieties is high, with an average degree of polymorphism of 98% and mean Shannon index, H0=7.50. Analysis on 18 Mangifera indica accessions also showed high degree of polymorphism of 99% and mean Shannon index, H0=5.74. Dice index of genetic similarity ranged from 0.0938 to 0.8046 among the Mangifera species. The dendrogram showed that the Mangifera species were grouped into three main divergent clusters. Cluster 1 comprised 14 accessions from Kedah and Perak. Cluster II and cluster III comprised 14 accessions from Sarawak and Sabah. Meanwhile, the Dice index of genetic similarity for 18 accessions of Mangifera indica ranged from 0.2588 to 0.7742. The dendrogram also showed the 18 accessions of Mangifera indica were grouped into three main clusters. Cluster I comprised 10 landraces of Mangifera indica from Kedah. Cluster II comprised 7 landraces of Mangifera indica followed by Chokanan to form Cluster III.

  18. Isozyme, ISSR and RAPD profiling of genotypes in marvel grass (Dichanthium annulatum).

    Science.gov (United States)

    Saxena, Raghvendra; Chandra, Amaresh

    2010-11-01

    Genetic analysis of 30 accessions of marvel grass (Dichanthium annulatum Forsk.), a tropical range grass collected from grasslands and open fields of drier regions, was carried out with the objectives of identifying unique materials that could be used in developing the core germplasm for such regions as well as to explore gene (s) for drought tolerance. Five inter-simple sequence repeat (ISSR) primers [(CA)4, (AGAC), (GACA) 4; 27 random amplified polymorphic DNA (RAPD) and four enzyme systems were employed in the present study. In total, ISSR yielded 61 (52 polymorphic), RAPD 269 (253 polymorphic) and enzyme 55 isozymes (44 polymorphic) bands. The average polymorphic information content (PIC) and marker index (MI) across all polymorphic bands of 3 markers systems ranged from 0.419 to 0.480 and 4.34 to 5.25 respectively Dendrogram analysis revealed three main clusters with all three markers. Four enzymes namely esterase (EST), polyphenoloxidase (PPO), peroxidase (PRX) and superoxide dismutase (SOD) revealed 55 alleles from a total of 16 enzyme-coding loci. Of these, 14 loci and 44 alleles were polymorphic. The mean number of alleles per locus was 3.43. Mean heterozygosity observed among the polymorphic loci ranged from 0.406 (SOD) to 0.836 (EST) and accession wise from 0.679 (1G3108) to 0.743 (IGKMD-10). Though there was intermixing of few accessions of one agro-climatic region to another largely groupings of accessions were with their regions of collections. Bootstrap analysis at 1000 iterations also showed large numbers of nodes (11 to 17) having strong clustering (> 50 bootstrap values) in all three marker systems. The accessions of the arid and drier regions forming one cluster are assigned as distinct core collection of Dichanthium and can be targeted for isolation of gene (s) for drought tolerance. Variations in isozyme allele numbers and high PIC (0.48) and MI (4.98) as observed with ISSR markers indicated their usefulness for germplasm characterization.

  19. Genomic evolutionary changes in Aegilops allopolyploids revealed by ISSR markers%山羊草属异源多倍体植物基因组进化的ISSR分析

    Institute of Scientific and Technical Information of China (English)

    龚汉雨; 刘爱华; 王建波

    2006-01-01

    使用31个ISSR引物对山羊草属Aegilops多倍体植物及其祖先二倍体(共23种)的基因组进行了分析,结果表明:与其二倍体祖先种相比,异源多倍体物种的基因组发生了很大变化.在含U基因组的异源多倍体物种中,U基因组相对而言变化很小,而其他基因组则发生了不同程度的变化.这表明当U基因组与其他基因组共存于多倍体物种中时,U基因组表现出较强的"同化效应".对这些基因组的进化进行了讨论.%Genomic evolutions were characterized by 31 ISSR (inter-simple sequence repeat)primers among 23 species in the genus Aegilops. The results indicate that the genome constituents of allopolyploid species had changed greatly through evolution compared with their ancestral diploid species. Genome U showed little alterations in U-containing allopolyploids, while others had undergone changes after allopolyploidization. In addition,genome U also displayed strong assimilation effect when it coexisted with other genomes, and the other genomes changed to varying degrees. The evolutionary changes of these genomes were discussed.

  20. Alu repeats as markers for forensic DNA analyses

    Energy Technology Data Exchange (ETDEWEB)

    Batzer, M.A.; Alegria-Hartman, M. [Lawrence Livermore National Lab., CA (United States); Kass, D.H. [Louisiana State Univ., New Orleans, LA (United States)] [and others

    1994-01-01

    The Human-Specific (HS) subfamily of Alu sequences is comprised of a group of 500 nearly identical members which are almost exclusively restricted to the human genome. Individual subfamily members share an average of 98.9% nucleotide identity with the HS subfamily consensus sequence, and have an average age of 2.8 million years. We have developed a Polymerase Chain Reaction (PCR) based assay using primers complementary to the 5 inch and 3 inch unique flanking DNA sequences from each HS Alu that allow the locus to be assayed for the presence or absence of the Alu repeat. The dimorphic HS Alu sequences probably inserted in the human genome after the radiation of modem humans (within the last 200,000-one million years) and represent a unique source of information for human population genetics and forensic DNA analyses. These sites can be developed into Dimorphic Alu Sequence Tagged Sites (DASTS) for the Human Genome Project. HS Alu family member insertions differ from other types of polymorphism (e.g. Variable Number of Tandem Repeat [VNTR] or Restriction Fragment Length Polymorphism [RFLP]) in that polymorphisms due to Alu insertions arise as a result of a unique event which has occurred only one time in the human population and spread through the population from that point. Therefore, individuals that share HS Alu repeats inherited these elements from a common ancestor. Most VNTR and RFLP polymorphisms may arise multiple times in parallel within a population.

  1. Analyses of the Genetic Relationships Among 22 Species of Manglietia Plants Using ISSR Markers%22种木莲属植物亲缘关系的ISSR分析

    Institute of Scientific and Technical Information of China (English)

    肖黎; 李晓玲; 王玉兵; 王希群; 陈发菊

    2011-01-01

    The genetic relationships among 22 species of Manglietia plants were analyzed by using inter-simple sequence repeat (ISSR) molecular-marked technique. Ten ISSR primers were selected to assess the genomes of 22 species of Manglietia plants. The result showed that a total of 595 DNA bands were amplified and 562 of which (94.45% ) were polymorphic. According to the Nei-Li genetic similarity of 0.68, UPGMA method cluster analysis indicated that these 22 species were classified into 6 cluster groups, and they were classified into 12 sub-cluster groups with the genetic similarity of 0. 70. The Nei-Li genetic similarities of 22 species of Manglietia ranged from 0.605 (between M. Yuyuanensis and M. Conifera) to 0. 956 (between M. Hookeri and M. Wangii). The average genetic similarity is 0.698, which suggests that Manglietia have a plenty of inter-species diversity. The genetic similarity coefficient between M. Hookeri and M. Wangii is 0. 956, and their genetic relationship is actually the closest. The result supports the classification of M. Patungensis, M. Yuyuanensis and M. Forrestii as independent species respectively.%利用ISSR标记分析技术探讨了22种木莲属植物的遗传亲缘关系.选用10对ISSR引物组合对木莲属22个种的基因组进行分析.结果表明:22个种共扩增出595条DNA带,其中多态性条带占94.45%;根据Nei-Li遗传相似性系数在0.68处,UPGMA聚类结果将22个种划分为6个类群,在0.70处进一步可分为12个亚类群;22种木莲属植物的Nei-Li遗传相似性系数变化范围为0.605(乳源木莲与球果木莲之间)~0.956(中缅木莲与滇南木莲之间),平均遗传相似性系数为0.698.说明木莲属植物种间差异性大.其中中缅木莲、滇南木莲之间遗传相似性系数为0.956,两者亲缘关系最近.该结果同时支持将巴东木莲、乳源木莲、滇桂木莲分别作为独立的种.

  2. Genetic Diversity and Similarity of Global Faba Bean(Vcia faba L.)Germplasm Revealed by ISSR Markers%世界蚕豆种质资源遗传多样性和相似性的ISSR分析

    Institute of Scientific and Technical Information of China (English)

    王海飞; 关建平; 孙雪莲; 马钰; 宗绪晓

    2011-01-01

    [Objective] The purpose of this research is to analyze the genetic diversity of global faba bean germplasm, to explore their genetic similarity and population structure, and to provide essential information for germplasm evaluation and effective utilization of faba bean genetic resources. [Method] The genetic similarity of 383 faba bean accessions from 35 countries was analyzed by using ISSR marker. [Result] Eleven ISSR primers generated 229 unambiguous bands, of which 212 were polymorphic,and the rate of polymorphic bands was 0.93. Gene diversity index (H) and allelic richness (NA) of different geographic groups of germplasm ranged from 0.16 to 0.28 and from 104 to 193, respectively. Chinese spring-seeding area group showed the highest genetic diversity (H = 0.28, NA = 193), and America group showed the lowest (H = 0.16, NA = 104). The spring faba bean germplasm was clearly separated from winter faba bean germplasm of China in UPGMA clustering analysis based on ISSR molecular marker data. Germplasm from China is quite distinct to that from exotic accessions. The accessions from Europe had a closer genetic similarity with that from North Africa. While the germplasm resources from Asia, Europe and Africa are closely related to their geographical distribution. [Conclusion ] Accessions from spring-seeding area of China were most diverse. Germplasm from America showed lowest diversity. The results indicated that the genetic similarity and diversity of faba bean germplasm are closely associated with their growth habit, their geographical origin and ecological distribution.%[目的]分析国内外蚕豆种质资源的遗传多样性,探索其遗传相似性和遗传结构,为世界蚕豆资源的综合评价和优良种质资源的发掘利用提供依据.[方法]利用ISSR标记技术,对来自世界35个国家的383份蚕豆资源的遗传相似性进行分析.[结果]筛选出的11条ISSR引物共扩增出229条条带,其中多态性条带212条(占93%).不同地理

  3. Analysis of genetic diversity and genetic relationship of wild hawthorn resources in Xinjiang by ISSR markers%新疆野生山楂资源遗传多样性及亲缘关系的ISSR分析

    Institute of Scientific and Technical Information of China (English)

    刘欢; 廖康; 刘娟; 赵世荣; 孙琪; 曹倩

    2016-01-01

    In order to provide some theoretical basis and technical support for protection and utilization of wild hawthorn resources, genetic diversity and genetic relationship of 63 samples from three varieties of wild hawthorn resources in Xinjiang were analyzed by ISSR molecular marker technology. The results showed that 233 bands were ampliifed with 12 primers by the ISSR-PCR ampliifcation, in which had 221 polymorphism bands, percent of polymorphism loci was 94.4%, and average 19.42 polymorphism bands were ampliifed by every primer. The results of analysis by the software of POPGENE1.32 showed that percentage of polymorphic loci, observed number of alleles, effective number of alleles, Nei’s genetic diversity and Shannon information index of Junggar hawthorn were higher than other two kinds of wild hawthorn, and the indexes of Crabapple hawthorn were lower than Junggar hawthorn and Altai hawthorn. The Nei’s genetic identities of three kinds of wild hawthorn were 0.862 2-0.904 4, and the average value was 0.890 2. The genetic distances of three kinds of wild hawthorn were 0.100 5-0.148 3, and the average value was 0.116 6. In the three kinds of wild hawthorn, genetic variation of Junggar hawthorn were more rich than the others, followed by Altai hawthorn, and Crabapple hawthorn had the less genetic variation. The genetic relationship between Altai hawthorn and Crabapple hawthorn was closer, but that was farther between Crabapple hawthorn and Junggar hawthorn.%为给新疆野生山楂资源的保护与利用提供理论依据和技术支持,采取新疆3种野生山楂共计63份材料,利用ISSR标记技术,对其遗传多样性和亲缘关系进行分析。结果表明,12条引物共扩增出233条带,其中多态性条带221条,多态性位点率为94.4%,平均每个引物获得19.42个扩增带。POPGENE1.32分析结果表明:3种野生山楂材料的多态性位点率、观测等位基因数、有效等位基因数、Nei’s基因多样性

  4. Alu repeats as markers for human population genetics

    Energy Technology Data Exchange (ETDEWEB)

    Batzer, M.A.; Alegria-Hartman, M. [Lawrence Livermore National Lab., CA (United States); Bazan, H. [Louisiana State Univ., New Orleans, LA (United States). Medical Center] [and others

    1993-09-01

    The Human-Specific (HS) subfamily of Alu sequences is comprised of a group of 500 nearly identical members which are almost exclusively restricted to the human genome. Individual subfamily members share an average of 97.9% nucleotide identity with each other and an average of 98.9% nucleotide identity with the HS subfamily consensus sequence. HS Alu family members are thought to be derived from a single source ``master`` gene, and have an average age of 2.8 million years. We have developed a Polymerase Chain Reaction (PCR) based assay using primers complementary to the 5 in. and 3 in. unique flanking DNA sequences from each HS Alu that allows the locus to be assayed for the presence or absence of an Alu repeat. Individual HS Alu sequences were found to be either monomorphic or dimorphic for the presence or absence of each repeat. The monomorphic HS Alu family members inserted in the human genome after the human/great ape divergence (which is thought to have occurred 4--6 million years ago), but before the radiation of modem man. The dimorphic HS Alu sequences inserted in the human genome after the radiation of modem man (within the last 200,000-one million years) and represent a unique source of information for human population genetics and forensic DNA analyses. These sites can be developed into Dimorphic Alu Sequence Tagged Sites (DASTS) for the Human Genome Project as well. HS Alu family member insertion dimorphism differs from other types of polymorphism (e.g. Variable Number of Tandem Repeat [VNTR] or Restriction Fragment Length Polymorphism [RFLP]) because individuals share HS Alu family member insertions based upon identity by descent from a common ancestor as a result of a single event which occurred one time within the human population. The VNTR and RFLP polymorphisms may arise multiple times within a population and are identical by state only.

  5. ISSR and isozyme characterization of androgenetic dihaploids reveals tetrasomic inheritance in tetraploid somatic hybrids between Solanum melongena and Solanum aethiopicum group Gilo.

    Science.gov (United States)

    Toppino, Laura; Mennella, Giuseppe; Rizza, Fulvia; D'Alessandro, Antonietta; Sihachakr, Darasinh; Rotino, Giuseppe L

    2008-01-01

    Gene exchanges between Solanum melongena and its allied relative Solanum aethiopicum are a crucial prerequisite for introgression of useful traits from the allied species into the cultivated eggplant. In order to evaluate the extent of genetic recombination between the 2 species, biochemical and molecular markers were employed. A dihaploid population obtained through anther culture of the corresponding tetraploid somatic hybrids was genetically analyzed. The extent of disomic/tetrasomic inheritance and segregation ratios of 3 isozyme systems and intersimple sequence repeat (ISSR) markers were evaluated. The dihaploids, being derived from microspores, allowed for simple, complete, and accurate analyses. The segregation of 280 ISSR markers (110 aethiopicum-specific, 104 melongena-specific, and 66 monomorphic) were evaluated in 71 dihaploids. According to the genetic constitution (simplex/duplex/triplex), almost 64% of the fragments revealed the tetrasomic and/or disomic inheritance. With regard to the assigned species-specific fragments, 68% and 4% were unambiguously the result of tetrasomic and disomic inheritance, respectively. Twenty-four of the 66 monomorphic ISSRs were inherited according to random chromatid segregation. The phenotypes of glucose-6-phosphate dehydrogenase (G-6-PDH), 6-phosphogluconate dehydrogenase (6-PGDH), and shikimate dehydrogenase (SKDH) were studied in 70 dihaploids and inferences were made about the allelic state of their 5 loci. The isozyme markers segregated in the dihaploids in a distorted manner, their segregations did not fit in with any of the expected segregation ratios. However, tetrasomic inheritance might be suggested for G-6-PDH 2 and SKDH 1 loci. Our results demonstrated that gene exchanges occurred readily in the somatic hybrids between S. melongena and S. aethiopicum gr. Gilo.

  6. Tandem repeat markers as novel diagnostic tools for high resolution fingerprinting of Wolbachia

    Directory of Open Access Journals (Sweden)

    Riegler Markus

    2012-01-01

    Full Text Available Abstract Background Strains of the endosymbiotic bacterium Wolbachia pipientis are extremely diverse both genotypically and in terms of their induced phenotypes in invertebrate hosts. Despite extensive molecular characterisation of Wolbachia diversity, little is known about the actual genomic diversity within or between closely related strains that group tightly on the basis of existing gene marker systems, including Multiple Locus Sequence Typing (MLST. There is an urgent need for higher resolution fingerprinting markers of Wolbachia for studies of population genetics, horizontal transmission and experimental evolution. Results The genome of the wMel Wolbachia strain that infects Drosophila melanogaster contains inter- and intragenic tandem repeats that may evolve through expansion or contraction. We identified hypervariable regions in wMel, including intergenic Variable Number Tandem Repeats (VNTRs, and genes encoding ankyrin (ANK repeat domains. We amplified these markers from 14 related Wolbachia strains belonging to supergroup A and were successful in differentiating size polymorphic alleles. Because of their tandemly repeated structure and length polymorphism, the markers can be used in a PCR-diagnostic multilocus typing approach, analogous to the Multiple Locus VNTR Analysis (MLVA established for many other bacteria and organisms. The isolated markers are highly specific for supergroup A and not informative for other supergroups. However, in silico analysis of completed genomes from other supergroups revealed the presence of tandem repeats that are variable and could therefore be useful for typing target strains. Conclusions Wolbachia genomes contain inter- and intragenic tandem repeats that evolve through expansion or contraction. A selection of polymorphic tandem repeats is a novel and useful PCR diagnostic extension to the existing MLST typing system of Wolbachia, as it allows rapid and inexpensive high-throughput fingerprinting of

  7. Simple sequence repeat marker development and genetic mapping in quinoa (Chenopodium quinoa Willd.)

    Indian Academy of Sciences (India)

    D. E. Jarvis; O. R. Kopp; E. N. Jellen; M. A. Mallory; J. Pattee; A. Bonifacio; C. E. Coleman; M. R. Stevens; D. J. Fairbanks; P. J. Maughan

    2008-04-01

    Quinoa is a regionally important grain crop in the Andean region of South America. Recently quinoa has gained international attention for its high nutritional value and tolerances of extreme abiotic stresses. DNA markers and linkage maps are important tools for germplasm conservation and crop improvement programmes. Here we report the development of 216 new polymorphic SSR (simple sequence repeats) markers from libraries enriched for GA, CAA and AAT repeats, as well as 6 SSR markers developed from bacterial artificial chromosome-end sequences (BES-SSRs). Heterozygosity (H) values of the SSR markers ranges from 0.12 to 0.90, with an average value of 0.57. A linkage map was constructed for a newly developed recombinant inbred lines (RIL) population using these SSR markers. Additional markers, including amplified fragment length polymorphisms (AFLPs), two 11S seed storage protein loci, and the nucleolar organizing region (NOR), were also placed on the linkage map. The linkage map presented here is the first SSR-based map in quinoa and contains 275 markers, including 200 SSR. The map consists of 38 linkage groups (LGs) covering 913 cM. Segregation distortion was observed in the mapping population for several marker loci, indicating possible chromosomal regions associated with selection or gametophytic lethality. As this map is based primarily on simple and easily-transferable SSR markers, it will be particularly valuable for research in laboratories in Andean regions of South America.

  8. Simple sequence repeat marker development and genetic mapping in quinoa (Chenopodium quinoa Willd.).

    Science.gov (United States)

    Jarvis, D E; Kopp, O R; Jellen, E N; Mallory, M A; Pattee, J; Bonifacio, A; Coleman, C E; Stevens, M R; Fairbanks, D J; Maughan, P J

    2008-04-01

    Quinoa is a regionally important grain crop in the Andean region of South America. Recently quinoa has gained international attention for its high nutritional value and tolerances of extreme abiotic stresses. DNA markers and linkage maps are important tools for germplasm conservation and crop improvement programmes. Here we report the development of 216 new polymorphic SSR (simple sequence repeats) markers from libraries enriched for GA, CAA and AAT repeats, as well as 6 SSR markers developed from bacterial artificial chromosome-end sequences (BES-SSRs). Heterozygosity (H) values of the SSR markers ranges from 0.12 to 0.90, with an average value of 0.57. A linkage map was constructed for a newly developed recombinant inbred lines (RIL) population using these SSR markers. Additional markers, including amplified fragment length polymorphisms (AFLPs), two 11S seed storage protein loci, and the nucleolar organizing region (NOR), were also placed on the linkage map. The linkage map presented here is the first SSR-based map in quinoa and contains 275 markers, including 200 SSR. The map consists of 38 linkage groups (LGs) covering 913 cM. Segregation distortion was observed in the mapping population for several marker loci, indicating possible chromosomal regions associated with selection or gametophytic lethality. As this map is based primarily on simple and easily-transferable SSR markers, it will be particularly valuable for research in laboratories in Andean regions of South America.

  9. Development of simple sequence repeat markers and diversity analysis in alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Wang, Zan; Yan, Hongwei; Fu, Xinnian; Li, Xuehui; Gao, Hongwen

    2013-04-01

    Efficient and robust molecular markers are essential for molecular breeding in plant. Compared to dominant and bi-allelic markers, multiple alleles of simple sequence repeat (SSR) markers are particularly informative and superior in genetic linkage map and QTL mapping in autotetraploid species like alfalfa. The objective of this study was to enrich SSR markers directly from alfalfa expressed sequence tags (ESTs). A total of 12,371 alfalfa ESTs were retrieved from the National Center for Biotechnology Information. Total 774 SSR-containing ESTs were identified from 716 ESTs. On average, one SSR was found per 7.7 kb of EST sequences. Tri-nucleotide repeats (48.8 %) was the most abundant motif type, followed by di-(26.1 %), tetra-(11.5 %), penta-(9.7 %), and hexanucleotide (3.9 %). One hundred EST-SSR primer pairs were successfully designed and 29 exhibited polymorphism among 28 alfalfa accessions. The allele number per marker ranged from two to 21 with an average of 6.8. The PIC values ranged from 0.195 to 0.896 with an average of 0.608, indicating a high level of polymorphism of the EST-SSR markers. Based on the 29 EST-SSR markers, assessment of genetic diversity was conducted and found that Medicago sativa ssp. sativa was clearly different from the other subspecies. The high transferability of those EST-SSR markers was also found for relative species.

  10. Assessment of genetic diversity in indigenous turmeric (Curcuma longa) germplasm from India using molecular markers.

    Science.gov (United States)

    Verma, Sushma; Singh, Shweta; Sharma, Suresh; Tewari, S K; Roy, R K; Goel, A K; Rana, T S

    2015-04-01

    Curcuma longa L., commonly known as turmeric, is one of the economically and medicinally important plant species. It is predominantly cultivated in the tropical and sub tropical countries. India is the largest producer, and exporter of turmeric in the world, followed by China, Indonesia, Bangladesh and Thailand. In the present study, Directed Amplification of Minisatellite DNA (DAMD) and Inter Simple Sequence Repeats (ISSR), methods were used to estimate the genetic variability in indigenous turmeric germplasm. Cumulative data analysis for DAMD (15) and ISSR (13) markers resulted into 478 fragments, out of which 392 fragments were polymorphic, revealing 82 % polymorphism across the turmeric genotypes. Wide range of pairwise genetic distances (0.03-0.59) across the genotypes revealed that these genotypes are genetically quite diverse. The UPGMA dendrogram generated using cumulative data showed significant relationships amongst the genotypes. All 29 genotypes studied grouped into two clusters irrespective of their geographical affiliations with 100 % bootstrap value except few genotypes, suggesting considerable diversity amongst the genotypes. These results suggested that the current collection of turmeric genotypes preserve the vast majority of natural variations. The results further demonstrate the efficiency and reliability of DAMD and ISSR markers in determining the genetic diversity and relationships among the indigenous turmeric germplasm. DAMD and ISSR profiling have identified diverse turmeric genotypes, which could be further utilized in various genetic improvement programmes including conventional as well as marker assisted breeding towards development of new and desirable turmeric genotypes.

  11. Population structure and genotypic variation of Crataegus pontica inferred by molecular markers.

    Science.gov (United States)

    Rahmani, Mohammad-Shafie; Shabanian, Naghi; Khadivi-Khub, Abdollah; Woeste, Keith E; Badakhshan, Hedieh; Alikhani, Leila

    2015-11-01

    Information about the natural patterns of genetic variability and their evolutionary bases are of fundamental practical importance for sustainable forest management and conservation. In the present study, the genetic diversity of 164 individuals from fourteen natural populations of Crataegus pontica K.Koch was assessed for the first time using three genome-based molecular techniques; inter-retrotransposon amplified polymorphism (IRAP); inter-simple sequence repeats (ISSR) and start codon targeted (SCoT) polymorphism. IRAP, ISSR and SCoT analyses yielded 126, 254 and 199 scorable amplified bands, respectively, of which 90.48, 93.37 and 83.78% were polymorphic. ISSR revealed efficiency over IRAP and SCoT due to high effective multiplex ratio, marker index and resolving power. The dendrograms based on the markers used and combined data divided individuals into three major clusters. The correlation between the coefficient matrices for the IRAP, ISSR and SCoT data was significant. A higher level of genetic variation was observed within populations than among populations based on the markers used. The lower divergence levels depicted among the studied populations could be seen as evidence of gene flow. The promotion of gene exchange will be very beneficial to conserve and utilize the enormous genetic variability. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. A blackberry (Rubus L.) expressed sequence tag library for the development of simple sequence repeat markers

    Science.gov (United States)

    A blackberry (Rubus L.) expressed sequence tag (EST) library was produced for developing simple sequence repeat (SSR) markers from the tetraploid blackberry cultivar, Merton Thornless, the source of the thornless trait in commercial cultivars. RNA was extracted from young expanding leaves and used f...

  13. Development and characterization of highly polymorphic long TC repeat microsatellite markers for genetic analysis of peanut

    Directory of Open Access Journals (Sweden)

    Macedo Selma E

    2012-02-01

    Full Text Available Abstract Background Peanut (Arachis hypogaea L. is a crop of economic and social importance, mainly in tropical areas, and developing countries. Its molecular breeding has been hindered by a shortage of polymorphic genetic markers due to a very narrow genetic base. Microsatellites (SSRs are markers of choice in peanut because they are co-dominant, highly transferrable between species and easily applicable in the allotetraploid genome. In spite of substantial effort over the last few years by a number of research groups, the number of SSRs that are polymorphic for A. hypogaea is still limiting for routine application, creating the demand for the discovery of more markers polymorphic within cultivated germplasm. Findings A plasmid genomic library enriched for TC/AG repeats was constructed and 1401 clones sequenced. From the sequences obtained 146 primer pairs flanking mostly TC microsatellites were developed. The average number of repeat motifs amplified was 23. These 146 markers were characterized on 22 genotypes of cultivated peanut. In total 78 of the markers were polymorphic within cultivated germplasm. Most of those 78 markers were highly informative with an average of 5.4 alleles per locus being amplified. Average gene diversity index (GD was 0.6, and 66 markers showed a GD of more than 0.5. Genetic relationship analysis was performed and corroborated the current taxonomical classification of A. hypogaea subspecies and varieties. Conclusions The microsatellite markers described here are a useful resource for genetics and genomics in Arachis. In particular, the 66 markers that are highly polymorphic in cultivated peanut are a significant step towards routine genetic mapping and marker-assisted selection for the crop.

  14. Population Genetic Structure and Marker Trait Associations Using Morphological, Phytochemical and Molecular Parameters in Habenaria edgeworthii-a Threatened Medicinal Orchid of West Himalaya, India.

    Science.gov (United States)

    Giri, Lalit; Jugran, Arun Kumar; Bahukhandi, Amit; Dhyani, Praveen; Bhatt, Indra D; Rawal, Ranbeer Singh; Nandi, Shyamal Kumar; Dhar, Uppeandra

    2017-01-01

    Habenaria edgeworthii Hook. f. ex Collett is an important terrestrial orchid used in different Ayurvedic formulations. In the present study, variations among morphological, phytochemical and molecular markers were assessed. A significant difference was observed among populations using morphological traits. Inter-simple sequence repeat (ISSR) data revealed lower genetic diversity at population level (He = 0.207) as compared to species level (He = 0.334). Analysis of molecular variance (AMOVA) indicates 74 % variation among populations and 26 % within population. Tuber extracts showed significantly (p morphological and phytochemical attributes were studied using multiple regression analysis (MRA). Several ISSR fragments were associated with some morphological and phytochemical traits. These ISSR fragments can be useful for breeding programme of the species when no other genetic information, such as linkage maps and quantitative trait loci, is available.

  15. 园艺作物的ISSR分子标记研究及应用%Inter-simple sequence repeat and its application in horticultural crop research

    Institute of Scientific and Technical Information of China (English)

    刘淑芹; 吴凤芝; 刘守伟

    2012-01-01

    ISSR (Inter-simple sequence repeat)是一种基于微卫星序列发展起来的新型分子标记方法,具有无需知道任何靶标序列的微卫星背景信息、遗传多态性高、稳定高效、检测快速等特点.目前,ISSR分子标记技术在园艺作物的遗传多样性研究、品种鉴定、遗传图谱构建、基因定位及分子标记辅助育种等方面得到了广泛应用,文章就ISSR标记的原理、方法、特点及其在园艺作物研究中的应用加以综述.%Inter-simple sequence repeat (ISSR) is a new molecular marker method which is based on micro-satellite technique with advantages of simple, quick, stable, reliable and higher DNA polymorphism, etc. At present, ISSR has been widely applied in genetic diversity research, genetic map construction, genetic mapping, molecular marker assisted breeding and variety purity identification of horticultural crops. The purpose of this review is to introduce principles, methods, characteristics of the ISSR and its application in the research of horticultural crops.

  16. Genetic Diversity of 29 H yac inth Germplasm Resources Revealed by Using ISSR Markers%利用ISSR分子标记分析29个风信子品种的遗传多样性

    Institute of Scientific and Technical Information of China (English)

    胡凤荣; 胡月苗; 王斐; 任翠

    2015-01-01

    现代栽培的所有园艺风信子都是由原种风信子(Hyacinth orientalis)培育驯化而来,其遗传基础相对较窄,但经过几个世纪栽培选育,已经发生很大变化。为研究风信子品种之间的遗传关系,本研究利用12条引物对29个风信子品种进行ISSR分子标记研究,扩增获得109条谱带,多态性条带有103条,占条带总数的94.5%,表明风信子具有较高的遗传多样性。29个风信子品种的遗传距离范围为0.0185~0.8201,平均遗传距离为0.4498。其中‘Atlantic’与‘Gipsy Queen’遗传距离最大为0.8201,亲缘关系最远‘;Blue star’与‘Delf Blue’遗传距离最小,两品种之间相似度较大,亲缘关系最近。使用软件NTSYS-pc (2.10e版)获得UPGMA聚类树形图,以相似系数0.635为阈值,29个风信子品种可聚为两大类。聚类分析发现相同色系的品种几乎聚为一类,说明同色系风信子品种亲缘关系较近。因此,在杂交育种时可选择不同花色的品种作亲本,进行新品种的选育。%All modern horticultural hyacinths are domesticated by the protospecies-Hyacinth orientalis. Though the protospecies have relatively narrow genetic basis, they have had great changes due to cultivation and breeding for centuries. In order to study the genetic relationship between varieties of hyacinth, 12 primers of ISSR molecular markers were used to analyze 29 species of H yac inth which generated 109 bands, including 103 polymorphism bands account for 94.5% of the total. Each pair of primers can amplify nine bands in average. The genetic distances among 29 Hyacinths orientalis were from 0.018 5 to 0.820 1 with the average of 0.449 8. There is the longest genetic distance of 0.820 1 between‘Atlantic’and‘Gipsy Queen’. Whereas between‘Blue star’and‘Delf Blue’being the shortest genetic distance. NTSYS-PC (Version 2.10e) was applied to UPGMA clustering analysis to construct the phylogenetic

  17. Analysis of simple sequence repeats markers derived from Phytophthora sojae expressed sequence tags

    Institute of Scientific and Technical Information of China (English)

    ZHU Zhendong; HUO Yunlong; WANG Xiaoming; HUANG Junbin; WU Xiaofei

    2004-01-01

    Five thousand and eight hundred publicly available expressed sequence tags (ESTs) of Phytophthora sojae were electronically searched and 415 simple sequence repeats (SSRs) were identified in 369 ESTs. The average density of SSRs was one SSR per 8.9 kb of EST sequence screened. The most frequent repeats were trinucleotide repeats (50.1%) and the least frequent were tetranucleotide repeats (8.2%). Forty primer pairs were designed and tested on 5 strains of P. sojae. Thirty-three primer pairs had successful PCR amplifications. Of the 33 functional primer pairs, 28 primer pairs produced characteristic SSR bands of the expected size, and 15 primer pairs (45.5%) detected polymorphism among 5 tested strains of P. sojae. Based on the polymorphisms detected with 20 EST-SSR markers, the 5 tested strains of P. sojae were clustered into 3 groups. In this study, the SSR markers of P. sojae were developed for the first time. These markers could be useful for identification, genetic variation study, and molecular mapping of P. sojae and its relative species.

  18. Development of simple sequence repeat (SSR) markers of sesame (Sesamum indicum) from a genome survey.

    Science.gov (United States)

    Wei, Xin; Wang, Linhai; Zhang, Yanxin; Qi, Xiaoqiong; Wang, Xiaoling; Ding, Xia; Zhang, Jing; Zhang, Xiurong

    2014-04-22

    Sesame (Sesamum indicum), an important oil crop, is widely grown in tropical and subtropical regions. It provides part of the daily edible oil allowance for almost half of the world's population. A limited number of co-dominant markers has been developed and applied in sesame genetic diversity and germplasm identity studies. Here we report for the first time a whole genome survey used to develop simple sequence repeat (SSR) markers and to detect the genetic diversity of sesame germplasm. From the initial assembled sesame genome, 23,438 SSRs (≥5 repeats) were identified. The most common repeat motif was dinucleotide with a frequency of 84.24%, followed by 13.53% trinucleotide, 1.65% tetranucleotide, 0.3% pentanucleotide and 0.28% hexanucleotide motifs. From 1500 designed and synthesised primer pairs, 218 polymorphic SSRs were developed and used to screen 31 sesame accessions that from 12 countries. STRUCTURE and phylogenetic analyses indicated that all sesame accessions could be divided into two groups: one mainly from China and another from other countries. Cluster analysis classified Chinese major sesame varieties into three groups. These novel SSR markers are a useful tool for genetic linkage map construction, genetic diversity detection, and marker-assisted selective sesame breeding.

  19. Genome wide characterization of short tandem repeat markers in sweet orange (Citrus sinensis.

    Directory of Open Access Journals (Sweden)

    Manosh Kumar Biswas

    Full Text Available Sweet orange (Citrus sinensis is one of the major cultivated and most-consumed citrus species. With the goal of enhancing the genomic resources in citrus, we surveyed, developed and characterized microsatellite markers in the ≈347 Mb sequence assembly of the sweet orange genome. A total of 50,846 SSRs were identified with a frequency of 146.4 SSRs/Mbp. Dinucleotide repeats are the most frequent repeat class and the highest density of SSRs was found in chromosome 4. SSRs are non-randomly distributed in the genome and most of the SSRs (62.02% are located in the intergenic regions. We found that AT-rich SSRs are more frequent than GC-rich SSRs. A total number of 21,248 SSR primers were successfully developed, which represents 89 SSR markers per Mb of the genome. A subset of 950 developed SSR primer pairs were synthesized and tested by wet lab experiments on a set of 16 citrus accessions. In total we identified 534 (56.21% polymorphic SSR markers that will be useful in citrus improvement. The number of amplified alleles ranges from 2 to 12 with an average of 4 alleles per marker and an average PIC value of 0.75. The newly developed sweet orange primer sequences, their in silico PCR products, exact position in the genome assembly and putative function are made publicly available. We present the largest number of SSR markers ever developed for a citrus species. Almost two thirds of the markers are transferable to 16 citrus relatives and may be used for constructing a high density linkage map. In addition, they are valuable for marker-assisted selection studies, population structure analyses and comparative genomic studies of C. sinensis with other citrus related species. Altogether, these markers provide a significant contribution to the citrus research community.

  20. Genome wide characterization of short tandem repeat markers in sweet orange (Citrus sinensis).

    Science.gov (United States)

    Biswas, Manosh Kumar; Xu, Qiang; Mayer, Christoph; Deng, Xiuxin

    2014-01-01

    Sweet orange (Citrus sinensis) is one of the major cultivated and most-consumed citrus species. With the goal of enhancing the genomic resources in citrus, we surveyed, developed and characterized microsatellite markers in the ≈347 Mb sequence assembly of the sweet orange genome. A total of 50,846 SSRs were identified with a frequency of 146.4 SSRs/Mbp. Dinucleotide repeats are the most frequent repeat class and the highest density of SSRs was found in chromosome 4. SSRs are non-randomly distributed in the genome and most of the SSRs (62.02%) are located in the intergenic regions. We found that AT-rich SSRs are more frequent than GC-rich SSRs. A total number of 21,248 SSR primers were successfully developed, which represents 89 SSR markers per Mb of the genome. A subset of 950 developed SSR primer pairs were synthesized and tested by wet lab experiments on a set of 16 citrus accessions. In total we identified 534 (56.21%) polymorphic SSR markers that will be useful in citrus improvement. The number of amplified alleles ranges from 2 to 12 with an average of 4 alleles per marker and an average PIC value of 0.75. The newly developed sweet orange primer sequences, their in silico PCR products, exact position in the genome assembly and putative function are made publicly available. We present the largest number of SSR markers ever developed for a citrus species. Almost two thirds of the markers are transferable to 16 citrus relatives and may be used for constructing a high density linkage map. In addition, they are valuable for marker-assisted selection studies, population structure analyses and comparative genomic studies of C. sinensis with other citrus related species. Altogether, these markers provide a significant contribution to the citrus research community.

  1. Characterization and compilation of polymorphic simple sequence repeat (SSR markers of peanut from public database

    Directory of Open Access Journals (Sweden)

    Zhao Yongli

    2012-07-01

    Full Text Available Abstract Background There are several reports describing thousands of SSR markers in the peanut (Arachis hypogaea L. genome. There is a need to integrate various research reports of peanut DNA polymorphism into a single platform. Further, because of lack of uniformity in the labeling of these markers across the publications, there is some confusion on the identities of many markers. We describe below an effort to develop a central comprehensive database of polymorphic SSR markers in peanut. Findings We compiled 1,343 SSR markers as detecting polymorphism (14.5% within a total of 9,274 markers. Amongst all polymorphic SSRs examined, we found that AG motif (36.5% was the most abundant followed by AAG (12.1%, AAT (10.9%, and AT (10.3%.The mean length of SSR repeats in dinucleotide SSRs was significantly longer than that in trinucleotide SSRs. Dinucleotide SSRs showed higher polymorphism frequency for genomic SSRs when compared to trinucleotide SSRs, while for EST-SSRs, the frequency of polymorphic SSRs was higher in trinucleotide SSRs than in dinucleotide SSRs. The correlation of the length of SSR and the frequency of polymorphism revealed that the frequency of polymorphism was decreased as motif repeat number increased. Conclusions The assembled polymorphic SSRs would enhance the density of the existing genetic maps of peanut, which could also be a useful source of DNA markers suitable for high-throughput QTL mapping and marker-assisted selection in peanut improvement and thus would be of value to breeders.

  2. Genome Wide Characterization of Short Tandem Repeat Markers in Sweet Orange (Citrus sinensis)

    OpenAIRE

    BISWAS, Manosh Kumar; XU, Qiang; Mayer, Christoph; Deng, Xiuxin

    2014-01-01

    Sweet orange (Citrus sinensis) is one of the major cultivated and most-consumed citrus species. With the goal of enhancing the genomic resources in citrus, we surveyed, developed and characterized microsatellite markers in the ≈347 Mb sequence assembly of the sweet orange genome. A total of 50,846 SSRs were identified with a frequency of 146.4 SSRs/Mbp. Dinucleotide repeats are the most frequent repeat class and the highest density of SSRs was found in chromosome 4. SSRs are non-randomly dist...

  3. Genome Wide Characterization of Short Tandem Repeat Markers in Sweet Orange (Citrus sinensis)

    OpenAIRE

    Biswas, Manosh Kumar; Xu, Qiang; Mayer, Christoph; Deng, Xiuxin

    2014-01-01

    Sweet orange (Citrus sinensis) is one of the major cultivated and most-consumed citrus species. With the goal of enhancing the genomic resources in citrus, we surveyed, developed and characterized microsatellite markers in the ≈347 Mb sequence assembly of the sweet orange genome. A total of 50,846 SSRs were identified with a frequency of 146.4 SSRs/Mbp. Dinucleotide repeats are the most frequent repeat class and the highest density of SSRs was found in chromosome 4. SSRs are non-randomly dist...

  4. Genetic differentiation induced by spaceflight treatment of Cistanche deserticola and identification of inter-simple sequence repeat markers associated with its medicinal constituent contents

    Science.gov (United States)

    Wu, Y.; Yang, D. Y.; Tu, P. F.; Tian, Y. Z.; Guo, Y. H.; Wang, X. M.; Li, X. B.

    2011-02-01

    The dried, fleshy stems of Cistanche deserticola (Orobanchaceae) are popular tonics in Traditional Chinese Medicine (TCM) to treat the inability of kidney in expelling extra fluid in the body, causing fluid retention, and reform reproductive system. However, the wild plants of C. deserticola have become endangered due to habitat downsizing and over-harvesting for its medicinal usages. The present research was carried out for the following purposes: (1) promoting the space-breeding research; (2) providing molecular evidence for agricultural selective breeding; and (3) protecting this endangered herbal medicine and conserving its genetic resources.In this study, plants were cultivated from seeds specifically treated in spaceflight for seven days, and sampled to screen positive mutants and identify ISSR markers associated with their medicinal constituents. As a result, nine out of the 94 ISSR primers were showed high polymorphism, and a total of 118 bands were generated, of which 80 were polymorphic, ranging from 250 to 2600 bp. The spaceflight mutants were found to have lower coefficient of gene differentiation (Gst = 0.0269), and higher gene flow (Nm = 18.0740) than those of the controls (Gst = 0.2067 and Nm = 1.9185). The results demonstrated that most of the genetic variation were harnessed within populations (>97%). The Analysis of Molecular Variance (AMOVA) revealed high genetic variation within populations (88.03%) and low genetic differentiation among regions (-18.83%) and populations (30.79%), respectively. The results also indicated a profound difference between spaceflight condition and that on the earth. The unique vacuum environment of spaceflight was suggested to induce DNA mutation and various variations of C. deserticola. In addition, six particular ISSR markers were identified, cloned and sequenced; one of them, CA41939-934, was found positively correlated with acteoside with correlation coefficient values of 0.264 (P molecular evidence for

  5. Genetic Diversity and Structure of Lolium Species Surveyed on Nuclear Simple Sequence Repeat and Cytoplasmic Markers

    Directory of Open Access Journals (Sweden)

    Hongwei Cai

    2017-04-01

    Full Text Available To assess the genetic diversity and population structure of Lolium species, we used 32 nuclear simple sequence repeat (SSR markers and 7 cytoplasmic gene markers to analyze a total of 357 individuals from 162 accessions of 9 Lolium species. This survey revealed a high level of polymorphism, with an average number of alleles per locus of 23.59 and 5.29 and an average PIC-value of 0.83 and 0.54 for nuclear SSR markers and cytoplasmic gene markers, respectively. Analysis of molecular variance (AMOVA revealed that 16.27 and 16.53% of the total variation was due to differences among species, with the remaining 56.35 and 83.47% due to differences within species and 27.39 and 0% due to differences within individuals in 32 nuclear SSR markers set and 6 chloroplast gene markers set, respectively. The 32 nuclear SSR markers detected three subpopulations among 357 individuals, whereas the 6 chloroplast gene markers revealed three subpopulations among 160 accessions in the STRUCTURE analysis. In the clustering analysis, the three inbred species clustered into a single group, whereas the outbreeding species were clearly divided, especially according to nuclear SSR markers. In addition, almost all Lolium multiflorum populations were clustered into group C4, which could be further divided into three subgroups, whereas Lolium perenne populations primarily clustered into two groups (C2 and C3, with a few lines that instead grouped with L. multiflorum (C4 or Lolium rigidum (C6. Together, these results will useful for the use of Lolium germplasm for improvement and increase the effectiveness of ryegrass breeding.

  6. A blackberry (Rubus L. expressed sequence tag library for the development of simple sequence repeat markers

    Directory of Open Access Journals (Sweden)

    Main Dorrie S

    2008-06-01

    Full Text Available Abstract Background The recent development of novel repeat-fruiting types of blackberry (Rubus L. cultivars, combined with a long history of morphological marker-assisted selection for thornlessness by blackberry breeders, has given rise to increased interest in using molecular markers to facilitate blackberry breeding. Yet no genetic maps, molecular markers, or even sequences exist specifically for cultivated blackberry. The purpose of this study is to begin development of these tools by generating and annotating the first blackberry expressed sequence tag (EST library, designing primers from the ESTs to amplify regions containing simple sequence repeats (SSR, and testing the usefulness of a subset of the EST-SSRs with two blackberry cultivars. Results A cDNA library of 18,432 clones was generated from expanding leaf tissue of the cultivar Merton Thornless, a progenitor of many thornless commercial cultivars. Among the most abundantly expressed of the 3,000 genes annotated were those involved with energy, cell structure, and defense. From individual sequences containing SSRs, 673 primer pairs were designed. Of a randomly chosen set of 33 primer pairs tested with two blackberry cultivars, 10 detected an average of 1.9 polymorphic PCR products. Conclusion This rate predicts that this library may yield as many as 940 SSR primer pairs detecting 1,786 polymorphisms. This may be sufficient to generate a genetic map that can be used to associate molecular markers with phenotypic traits, making possible molecular marker-assisted breeding to compliment existing morphological marker-assisted breeding in blackberry.

  7. Diferenciación morfológica y por ISSR (Inter simple sequence repeats de especies del género Plukenetia (Euphorbiaceae de la Amazonía peruana: propuesta de una nueva especie

    Directory of Open Access Journals (Sweden)

    Ángel Rodríguez

    2011-05-01

    Full Text Available En el presente trabajo se estudian cinco especies del género Plukenetia de la Amazonía peruana: P. brachybotrya, P. loretensis, P. polyadenia, P. huayllabambana, P. volubilis (procedencia San Martín; y de un supuesto morfotipo (P. volubilis, procedencia Cusco. Los 126 especímenes estudiados fueron identificados mediante claves de caracteres morfológicos (formas de hojas, tallos y semilla; posición de glándulas basilaminares y posteriormente mediante marcadores moleculares ISSR (CAA, CAG, GACA. Los análisis morfológicos permitieron separar las cinco especies descritas: P. brachybotrya, P. loretensis, P. polyadenia, P. volubilis y P. huayllabambana. Los dos supuestos morfotipos de P. volubilis fueron discriminados por la posición de las glándulas, tamaño de semillas y forma del tallo. Los resultados proporcionados por el Análisis Factorial de Correspondencia (AFC y corroborados por el Índice de fijación (FST, distancia genética y el dendograma estimado por el método UPGMA, evidencian una fuerte diferenciación entre los seis taxa, corroborando la identidad taxonómica molecular de las cinco especies ya descritas morfológicamente. Además, los resultados (Fst y la distancia genética indicarian que P. volubilis (del Cusco podría ser una nueva especie de Sacha Inchi, aún no descrita para la ciencia.

  8. ISSR

    African Journals Online (AJOL)

    STORAGESEVER

    2009-10-05

    Oct 5, 2009 ... propagation and management of the plant is easy during .... no.4, no.7 and no.8 at a GS coefficient 0.77; the fourth sub group consisted ..... Li Q, Liu QC, Zhai H, Ma DF, Wang X, Li XQ, Wang YP (2008). Genetic ... Two new C-.

  9. Human Short Tandem Repeat (STR Markers for Paternity Testing in Pig-Tailed Macaques

    Directory of Open Access Journals (Sweden)

    DYAH PERWITASARI-FARAJALLAH

    2007-06-01

    Full Text Available This study investigated the use of human short tandem repeat (STR or microsatellite loci markers for assessing paternity and genetic structure of pig-tailed macaques (Macaca nemestrina breeding colony. Four human microsatellite primer pairs located at human map position D1S548, D3S1768, D5S820, and D2S1777, were amplified by polymerase chain reaction (PCR for pig-tailed macaques. Four loci were found to be clearly and reliably amplified, and three loci exhibited high levels of genetic heterogeneity. These loci were sufficiently informative to differentiate discretely between related and unrelated pairs.

  10. Automated discovery of single nucleotide polymorphism and simple sequence repeat molecular genetic markers.

    Science.gov (United States)

    Batley, Jacqueline; Jewell, Erica; Edwards, David

    2007-01-01

    Molecular genetic markers represent one of the most powerful tools for the analysis of genomes. Molecular marker technology has developed rapidly over the last decade, and two forms of sequence-based markers, simple sequence repeats (SSRs), also known as microsatellites, and single nucleotide polymorphisms (SNPs), now predominate applications in modern genetic analysis. The availability of large sequence data sets permits mining for SSRs and SNPs, which may then be applied to genetic trait mapping and marker-assisted selection. Here, we describe Web-based automated methods for the discovery of these SSRs and SNPs from sequence data. SSRPrimer enables the real-time discovery of SSRs within submitted DNA sequences, with the concomitant design of PCR primers for SSR amplification. Alternatively, users may browse the SSR Taxonomy Tree to identify predetermined SSR amplification primers for any species represented within the GenBank database. SNPServer uses a redundancy-based approach to identify SNPs within DNA sequence data. Following submission of a sequence of interest, SNPServer uses BLAST to identify similar sequences, CAP3 to cluster and assemble these sequences, and then the SNP discovery software autoSNP to detect SNPs and insertion/deletion (indel) polymorphisms.

  11. Establishment of the Optimum ISSR-PCR Reaction System in Hemerocallis spp.%大花萱草ISSR-PCR最佳反应体系的建立

    Institute of Scientific and Technical Information of China (English)

    曹冬梅; 康黎芳; 赵艳; 张超; 段九菊; 王云山

    2011-01-01

    Inter-simple sequence repeat (ISSR) method was applied to detect inter- and intra-specific genetic variation, and better reveal genetic relationship and genetic diversity. The factors which affect the inter-simple sequence repeat (ISSR) analysis were studied through 5 daylily cultivars. The optimal ISSR-PCR reaction conditions for daylily were established as follows: in a volume of 20 μ L containing 2 μ. L 10 x buffer, 0.2 mmol/L Dntp, 1.5 mmol/L Mg21, 0.5 mmol/L primer, 10.002 nkat Taq DNA polymerase, and 20 ng template DNA. The optimal amplified procedure was as follows: after a pre-denaturing of 5 min at 94 ℃, 40 cycles were performed with denaturing of 40 s at 94 ℃, annealing of 45 s at 50 ℃, extension of 1.5 min at 72 ℃, a final extension of 10 min at 72 ℃. The clear and stable amplified bands might be obtained with the above ISSR-PCR reaction conditions.%ISSR标记能揭示出种间、种内遗传变异情况,更好地揭示亲缘关系和遗传多样性.以大花萱草的5个品种为试材,研究了影响大花萱草ISSR-PCR扩增效果的各项因素,建立了适合大花萱草ISSR-PCR的最佳反应体系:20 μL反应体系中,10×buffer 2μL,dNTP 0.2 mmol/L,Mg2+浓度1.5 mmol/L,引物浓度0.5 mmol/L,7锄酶10.002 nkat,DNA模板20 ng.大花萱草的最佳ISSR扩增反应程序为:94℃预变性5 min;94℃变性40 s,50℃退火45 s,72℃延伸1.5 min,共40个循环;最后72℃延伸10 min.通过该反应程序进行的大花萱草ISSR扩增可获得清晰、稳定的条带.

  12. Simple sequence repeat markers useful for sorghum downy mildew (Peronosclerospora sorghi and related species

    Directory of Open Access Journals (Sweden)

    Odvody Gary N

    2008-11-01

    Full Text Available Abstract Background A recent outbreak of sorghum downy mildew in Texas has led to the discovery of both metalaxyl resistance and a new pathotype in the causal organism, Peronosclerospora sorghi. These observations and the difficulty in resolving among phylogenetically related downy mildew pathogens dramatically point out the need for simply scored markers in order to differentiate among isolates and species, and to study the population structure within these obligate oomycetes. Here we present the initial results from the use of a biotin capture method to discover, clone and develop PCR primers that permit the use of simple sequence repeats (microsatellites to detect differences at the DNA level. Results Among the 55 primers pairs designed from clones from pathotype 3 of P. sorghi, 36 flanked microsatellite loci containing simple repeats, including 28 (55% with dinucleotide repeats and 6 (11% with trinucleotide repeats. A total of 22 microsatellites with CA/AC or GT/TG repeats were the most abundant (40% and GA/AG or CT/TC types contribute 15% in our collection. When used to amplify DNA from 19 isolates from P. sorghi, as well as from 5 related species that cause downy mildew on other hosts, the number of different bands detected for each SSR primer pair using a LI-COR- DNA Analyzer ranged from two to eight. Successful cross-amplification for 12 primer pairs studied in detail using DNA from downy mildews that attack maize (P. maydis & P. philippinensis, sugar cane (P. sacchari, pearl millet (Sclerospora graminicola and rose (Peronospora sparsa indicate that the flanking regions are conserved in all these species. A total of 15 SSR amplicons unique to P. philippinensis (one of the potential threats to US maize production were detected, and these have potential for development of diagnostic tests. A total of 260 alleles were obtained using 54 microsatellites primer combinations, with an average of 4.8 polymorphic markers per SSR across 34

  13. High Genetic Diversity in a Rare, Narrowly Endemic Primrose Species: Primula interjacens by ISSR Analysis

    Institute of Scientific and Technical Information of China (English)

    XUEDa-Wei; GEXue-Jun; HAOGang; ZHANGChang-Qin

    2004-01-01

    Prirnula interjacens Chen (Primulaceae) is a rare and narrow endemic species of centralsouth of Yunnan Province in China. This species consists of two varieties: P.interjacens var. interjacens known with only one population, and P.interjacens var. epilosa with two populations. Intersimple sequence repeat (ISSR) marker was used to detect the genetic diversity of the three extant populations. We expected a low genetic diversity level, but our results revealed a high level of intraspecific genetic diversity (at population level: P=59.75%, HE=0.2368, and Hpop=0.3459; at species level: P=75.47%, HT= 0.320 5, and Hsp = 0.4618), probably resulting from floral heteromorphism and preferring outcrossing. A moderate level of genetic differentiation among populations was detected based on Nei's genetic diversity analysis (26.13%) and Shannon's diversity index (25.09%). Although P./ntedacens var. intedacens and P. interjacens var. epilosa were morphologically distinct, UPGMA cluster analysis showed that the two varieties had no distinct genetic differentiation and may be treated as a single taxon. Conservation measures are suggested, including in situ and ex situ strategies, based on the observed population genetic information.

  14. Research on Genetic Polymorphism of Cassava Germplasms and ISSR Molecular Markers%木薯种质资源遗传多态性ISSR分子标记的研究

    Institute of Scientific and Technical Information of China (English)

    彭靖茹; 甘志勇; 黎萍; 黄强; 杨金春; 石兰蓉; 付海天

    2012-01-01

    [目的]研究广西壮族自治区亚热带作物研究所保存的39份木薯种质资源遗传多样性.[方法]筛选出对木薯具有较好多态性和效果的ISSR引物,应用筛选出的引物对39份木薯种质资源进行PCR扩增,并对扩增条带进行统计和分析.同时根据品系间的遗传相似系数,利用UPGMA法进行聚类分析.[结果]筛选出10个ISSR引物,每个引物检测等位基因3~9个,平均为7个,获得70个扩增带形,扩增产物的片段大小范围在150~2 000 bp之间;聚类分析得到,在遗传相似系数0.67上将39个木薯品系分为2个类群,同时聚类分析结果表明,木薯之间的遗传距离基础非常狭窄,遗传相似系数大多在0.80~1.00之间.[结论]为我国优质木薯的育种奠定基础.%[Objective] This study aimed to analyze the genetic polymorphism of 39 cassava varieties (lines) saved in Guangxi Zhuang Autonomous Region Research Institute for Subtropical Crops. [Method] ISSR primers with good polymorphisms and amplification effects in cassava were selected for PCR amplification of 39 cassava germplasms, statistics and analysis of the amplified bands were conducted. In addition, cluster analysis was conducted by using UPGMA method based on the genetic similarity among cassava varieties (lines). [Result] 10 of ISSR primers were selected. A total of 70 clear electrophoretic bands were amplified, each primer had amplified 3-9 electrophoretic bands, with an average of 7, and the length of amplified bands ranged from 150 bp to 2 000 bp; cluster analysis showed that 39 cassava varieties (lines) were clustered into two categories by the similarity coefficient of 0.67; in addition, the genetic distance among cassava was very narrow, with genetic similarity coefficients ranged between 0. 80 and 1. 00. [ Conclusion ] This study laid a foundation for breeding of high-quality cassava in China.

  15. Analisis keragaman genetik manggis (Garcinia mangostana diiradiasi dengan sinar gamma berdasarkan penanda ISSR

    Directory of Open Access Journals (Sweden)

    ALFIN WIDIASTUTI

    2013-05-01

    Full Text Available Widiastuti A, Sobir, Suhartanto MR. 2013. Genetic variability analysis of mangosteen (Garcinia mangostana irradiated by gamma ray based on ISSR marker. Bioteknologi 10: 15-22. The aim of the research was to know the increasing of genetic variation within mangosteen based on banding pattern of ISSR marker as a response to some doses of irradiation gamma rays. Five dosages of irradiation gamma rays were 0 Gy, 20 Gy, 25 Gy, 30 Gy, 35 Gy and 40 Gy. The source of the seed that have been irradiated was from Cegal, Leuwiliang, Bogor. The results showed that gamma ray irradiation affecting mangosteen genetic changed. Because the randomize of gamma irradiation, it changed was individually. Gamma irradiation successfully increase genetic variability (5%.

  16. Simple sequence repeat marker associated with a natural leaf defoliation trait in tetraploid cotton.

    Science.gov (United States)

    Abdurakhmonov, I Y; Abdullaev, A A; Saha, S; Buriev, Z T; Arslanov, D; Kuryazov, Z; Mavlonov, G T; Rizaeva, S M; Reddy, U K; Jenkins, J N; Abdullaev, A; Abdukarimov, A

    2005-01-01

    Cotton (Gossypium hirsutum L.) leaf defoliation has a significant ecological and economical impact on cotton production. Thus the utilization of a natural leaf defoliation trait, which exists in wild diploid cotton species, in the development of tetraploid cultivated cotton will not only be cost effective, but will also facilitate production of very high-grade fiber. The primary goal of our research was to tag loci associated with natural leaf defoliation using microsatellite markers in Upland cotton. The F2 populations developed from reciprocal crosses between the two parental cotton lines--AN-Boyovut-2 (2n = 52), a late leaf defoliating type, and Listopad Beliy (2n = 52), a naturally early leaf defoliating type--demonstrated that the naturally early leaf defoliation trait has heritability values of 0.74 and 0.84 in the reciprocal F2 population. The observed phenotypic segregation difference in reciprocal crosses suggested a minor cytoplasmic effect in the phenotypic expression of the naturally early leaf defoliation trait. Results from the Kruskal-Wallis (KW) nonparametric test revealed that JESPR-13 (KW = 6.17), JESPR-153 (KW = 9.97), and JESPR-178 (KW = 13.45) Simple sequence repeat (SSR) markers are significantly associated with natural leaf defoliation in the mapping population having stable estimates at empirically obtained critical thresholds (P < .05-.0001). JESPR-178 revealed the highest estimates (P < .0001) for association with the natural leaf defoliation trait, exceeding maximum empirical threshold values. JESPR-178 was assigned to the short arm of chromosome 18, suggesting indirectly that genes associated with natural leaf defoliation might be located on this chromosome. This microsatellite marker may have the potential for use to introgress the naturally early leaf defoliation quantitative trait loci (QTL) from the donor line Listopad Beliy to commercial varieties of cotton through marker-assisted selection programs.

  17. Primary study of ISSR molecular markers for the leaf rust resistance gene Lr37 in wheat%小麦抗叶锈基因Lr37 ISSR分子标记的初步研究

    Institute of Scientific and Technical Information of China (English)

    张立荣; 徐大庆; 杨文香; 刘大群

    2004-01-01

    利用ISSR(内部简单重复序列)技术对感病品种Thatcher及20个以Thatcher为轮回亲本的小麦抗叶锈病近等基因系进行分析,找到1个与Lr37基因连锁的ISSR标记.经过多次重复发现,在一套ISSR引物中,引物UBC812在小麦抗叶锈基因Lr37近等基因系间表现多态性.当用这个引物对已知含Lr37基因的3个抗病材料及其它不含Lr37基因的感病材料进行检测时,多态性标记UBC812-1200可以从3个含Lr37基因的抗病材料中检测到1条1 200bp的多态性带,而在其它感病材料中均未出现.

  18. Condition optimization and primer screening for ISSR-PCR of Demodex%蠕形螨ISSR-PCR的反应体系优化及引物筛选

    Institute of Scientific and Technical Information of China (English)

    刘艳荣; 刘付红; 肖克源; 郭淑玲; 冯玉新; 袁方曙

    2012-01-01

    目的 以蠕形螨的DNA为模板,对蠕形螨简单重复序列锚定PCR (ISSR-PCR)反应体系优化并对ISSR引物进行筛选,为ISSR分析奠定基础.方法 用自然沉降法收集山羊蠕形螨,用基因组DNA提取试剂盒提取山羊蠕形螨DNA,并以此为模板,以(CA)8RG(R为1分子的嘧啶)为引物,利用正交实验法,从Mg2+浓度、引物用量、dNTPs、DNA模板用量和TaqDNA聚合酶以及退火温度对蠕形螨ISSR-PCR反应体系进行优化,并以优化的反应体系筛选ISSR-PCR的引物.结果 ISSR-PCR 25 μL最佳反应体系包括:0.4 mmol/L Mg2+、30 pmol引物、30~60 ng模板DNA、2u Taq酶,最佳退火温度为49℃、循环35次.筛选出10条条带清晰、多态性好、重复性高的引物.结论 筛选出蠕形螨ISSR-PCR最佳反应体系和理想的引物,为蠕形螨的分子生物学研究奠定了基础.%Objective To optimize conditions and screening for primers for inter-simple sequence repeat-anchored (IS-SR-PCR) of Demodex, using genome DNA as the template, preparing for the genetic diversity research of Demodex with ISSR mark. Methods After mites were rinsed and precipitated in double distilled water (DDW), the genome DNA of Demodex was extracted with a kit. Then experiments were carried out using DNA of Demodex as the template, and (CA) 8 RG as the primer. The orthogonal array was conducted, and the involved factors were as follows: concentrations of Mg2+ , dNTPs, primers, template DNA and Taq DNA polymerase and return temperature. An optimal reaction system of ISSR-PCR of Demodex was established, through which an optimal primer was obtained. Results An optimal 25 (xL of ISSR-PCR amplification system contained 2.0 mmol/L of Mg2+ , 0. 4 mmol/L of dNTPs, 30-60 ng of template DNA, 30 pmol of primers, and 2U of Taq DNA polymerase. The optional return temperature for the primer was 49 t, and the cycle index was 35. 10 primers were selected, with clear bands, good polymorphism and high repetition. Conclusion

  19. Genetic relationship of Curcuma species from Northeast India using PCR-based markers.

    Science.gov (United States)

    Das, Archana; Kesari, Vigya; Satyanarayana, Vinod M; Parida, Ajay; Rangan, Latha

    2011-09-01

    Molecular genetic fingerprints of nine Curcuma species from Northeast India were developed using PCR-based markers. The aim involves elucidating there intra- and inter-specific genetic diversity important for utilization, management, and conservation. Twelve random amplified polymorphic DNA (RAPD), 19 Inter simple sequence repeats (ISSRs), and four amplified fragment length polymorphism (AFLP) primers produced 266 polymorphic fragments. ISSR confirmed maximum polymorphism of 98.55% whereas RAPD and AFLP showed 93.22 and 97.27%, respectively. Marker index and polymorphic information content varied in the range of 8.64-48.1, 19.75-48.14, and 25-28 and 0.17-0.48, 0.19-0.48, and 0.25-0.29 for RAPD, ISSR, and AFLP markers, respectively. The average value of number of observed alleles, number of effective alleles, mean Nei's gene diversity, and Shannon's information index were 1.93-1.98, 1.37-1.62, 0.23-0.36, and 0.38-0.50, respectively, for three DNA markers used. Dendrograms based on three molecular data using unweighted pair group method with arithmetic mean (UPGMA) was congruent and classified the Curcuma species into two major clusters. Cophenetic correlation coefficient between dendrogram and original similarity matrix were significant for RAPD (r = 0.96), ISSR (r = 0.94), and AFLP (r = 0.97). Clustering was further supported by principle coordinate analysis. High genetic polymorphism documented is significant for conservation and further improvement of Curcuma species.

  20. Identification of a Simple Sequence Repeat molecular-marker set for large-scale analyses of pear germplasm

    Directory of Open Access Journals (Sweden)

    Gabriel Dequigiovanni

    2012-01-01

    Full Text Available Simple Sequence Repeats (SSR are molecular markers suitable to assess the genetic variation of germplasm resources; however, large-scale SSR use requires protocol optimization. The present work aimed to identify SSR markers, developed for pear and other fruit species that are effective in characterizing pear germplasm collections and in demonstrating their use in providing support for genetic breeding programs. From a total of 62 SSR markers investigated, 23 yielding reproducible and polymorphic patterns were used to genotype a sample of 42 pear accessions of the Brazilian Pear Germplasm Bank (PGB. When compared to these 23 SSR markers, a subset of eleven markers, selected based on He, PIC and PId, was used to distinguish individual accessions and perform cluster analysis with similar efficacy. Genetic diversity analysis clustered the European, Japanese and Chinese accessions in distinct groups. This markers subset constitutes a valuable tool for several applications related to pear genetic resources management and breeding.

  1. ISSR Analysis of Chlorophytum Treated by Three Kinds of Chemical Mutagen

    Institute of Scientific and Technical Information of China (English)

    WU Lijun; LI Muzi; YANG Xue; YANG Tao; WANG Jingang

    2011-01-01

    Chlorophytum leaves were treated with three kinds of chemical mutagen, EMS, DES and NaN3 with different concentration to obtain the variation materials with excellent properties. The results showed that the genetic similarity coefficient of variation plants with EMS treatment was between 0.648 and 0.868, with NaN3 treatment was between 0.598 and 0.859, and with DES treatment was between 0.668 and 0.904, of which the mutagenic effects with 0.8% EMS, 250 mg. L-1 NaN3 and 0.3% DES on chloro- phytum were the best. ISSR molecular marker technique was used to analyze their genetic diversities. Total 392 polymorphic bands were obtained through 18 ISSR primers. Polymorphic ratio was 72.4%, which showed that DNA mutation took place in various degrees

  2. Efficient development of highly polymorphic microsatellite markers based on polymorphic repeats in transcriptome sequences of multiple individuals.

    Science.gov (United States)

    Vukosavljev, M; Esselink, G D; van 't Westende, W P C; Cox, P; Visser, R G F; Arens, P; Smulders, M J M

    2015-01-01

    The first hurdle in developing microsatellite markers, cloning, has been overcome by next-generation sequencing. The second hurdle is testing to differentiate polymorphic from nonpolymorphic loci. The third hurdle, somewhat hidden, is that only polymorphic markers with a large effective number of alleles are sufficiently informative to be deployed in multiple studies. Both steps are laborious and still performed manually. We have developed a strategy in which we first screen reads from multiple genotypes for repeats that show the most length variants, and only these are subsequently developed into markers. We validated our strategy in tetraploid garden rose using Illumina paired-end transcriptome sequences of 11 roses. Of 48 tested two markers failed to amplify, but all others were polymorphic. Ten loci amplified more than one locus, indicating duplicated genes or gene families. Completely avoiding duplicated loci will be difficult because the range of numbers of predicted alleles of highly polymorphic single- and multilocus markers largely overlapped. Of the remainder, half were replicate markers (i.e. multiple primer pairs for one locus), indicating the difficulty of correctly filtering short reads containing repeat sequences. We subsequently refined the approach to eliminate multiple primer sets to the same loci. The remaining 18 markers were all highly polymorphic, amplifying on average 11.7 alleles per marker (range = 6-20) in 11 tetraploid roses, exceeding the 8.2 alleles per marker of the 24 most polymorphic markers genotyped previously. This strategy therefore represents a major step forward in the development of highly polymorphic microsatellite markers.

  3. Genetic Diversity of Landraces in Gossypium arboreum L. Race sinense Assessed with Simple Sequence Repeat Markers

    Institute of Scientific and Technical Information of China (English)

    Wang-Zhen Guo; Bao-Liang Zhou; Lu-Ming Yang; Wei Wang; Tian-Zhen Zhang

    2006-01-01

    Asiatic cotton (Gossypium arboreum L.) is an "Old World" cultivated cotton species, the sinense race of which is planted extensively in China. This species is still used in the current tetraploid cotton breeding program as an elite germplasm line, and is also used as a model for genomic research in Gossypium. In the present study, 60 cotton microsatellite markers, averaging 4.6 markers for each A-genome chromosome,were chosen to assess the genetic diversity of 109 accessions. These included 106 G. arboreum landraces,collected from 18 provinces throughout four Asiatic cotton-growing regions in China. A total of 128 alleles were detected, with an average of 2.13 alleles per locus. The largest number of alleles, as well as the maximum number of polymorphic loci, was detected in the A03 linkage group. No polymorphic alleles were detected on chromosome 10. The polymorphism information content for the 22 polymorphic microsatellite loci varied from 0.52 to 0.98, with an average of 0.89. Genetic diversity analysis revealed that the landraces in the Southern region had more genetic variability than those from the other two regions, and no significant difference was detected between landraces in the Yangtze and the Yellow River Valley regions. These findings are consistent with the history of sinense introduction, with the Southern region being the presumed center of origin for Chinese Asiatic cotton, and with subsequent northeastward extension to the Yangtze and Yellow River Valleys. Cluster analysis, based on simple sequence repeat data for 60 microsatellite loci, clearly differentiated Vietnamese and G. herbaceum landraces from the sinense landrace. No relationship between inter-variety similarity and geographical ecological region was observed. The present findings indicate that the Southern region landraces may have been directly introduced into the provinces in the middle and lower Yangtze River Valley, where Asiatic cotton was most extensively grown, and further race

  4. Genetic characterization of the gypsy moth from China (Lepidoptera, Lymantriidae using inter simple sequence repeats markers.

    Directory of Open Access Journals (Sweden)

    Fang Chen

    Full Text Available This study provides the first genetic characterization of the gypsy moth from China (Lymantriadispar, one of the most recognized pests of forests and ornamental trees in the world. We assessed genetic diversity and structure in eight geographic populations of gypsy moths from China using five polymorphic Inter simple sequence repeat markers, which produced reproducible banding patterns. We observed 102 polymorphic loci across the 176 individuals sampled. Overall genetic diversity (Nei's, H was 0.2357, while the mean genetic diversity within geographic populations was 0.1845 ± 0.0150. The observed genetic distance among the eight populations ranged from 0.0432 to 0.1034. Clustering analysis (using an unweighted pair-group method with arithmetic mean and multidimensional scaling, revealed strong concordance between the strength of genetic relationships among populations and their geographic proximity. Analysis of molecular variance demonstrated that 25.43% of the total variability (F ST = 0.2543, P < 0.001 was attributable to variation among geographic populations. The results of our analyses investigating the degree of polymorphism, genetic diversity (Nei's and Shannon and genetic structure, suggest that individuals from Hebei may be better able to adapt to different environments and to disperse to new habitats. This study provides crucial genetic information needed to assess the distribution and population dynamics of this important pest species of global concern.

  5. Genetic variability in maned wolf based on heterologous short-tandem repeat markers from domestic dog.

    Science.gov (United States)

    Salim, D C; Akimoto, A A; Carvalho, C B; Oliveira, S F; Grisolia, C K; Moreira, J R; Klautau-Guimarães, M N

    2007-06-20

    The maned wolf (Chrysocyon brachyurus) is the largest South American canid. Habitat loss and fragmentation, due to agricultural expansion and predatory hunting, are the main threats to this species. It is included in the official list of threatened wildlife species in Brazil, and is also protected by IUCN and CITES. Highly variable genetic markers such as microsatellites have the potential to resolve genetic relationships at all levels of the population structure (among individuals, demes or metapopulations) and also to identify the evolutionary unit for strategies for the conservation of the species. Tests were carried out to verify whether a class of highly polymorphic tetranucleotide repeats described for the domestic dog effectively amplifies DNA in the maned wolf. All five loci studied were amplified; however, one of these, was shown to be monomorphic in 69 maned wolf samples. The average allele number and estimated heterozygosity per polymorphic locus were 4.3 and 67%, respectively. The genetic variability found for this species, which is considered threatened with extinction, showed similar results when compared to studies of other canids.

  6. Transcriptome characterisation and simple sequence repeat marker discovery in the seagrass Posidonia oceanica

    Science.gov (United States)

    D’Esposito, D.; Orrù, L.; Dattolo, E.; Bernardo, L.; Lamontara, A.; Orsini, L.; Serra, I.A; Mazzuca, S.; Procaccini, G.

    2016-01-01

    Posidonia oceanica is an endemic seagrass in the Mediterranean Sea, where it provides important ecosystem services and sustains a rich and diverse ecosystem. P. oceanica meadows extend from the surface to 40 meters depth. With the aim of boosting research in this iconic species, we generated a comprehensive RNA-Seq data set for P. oceanica by sequencing specimens collected at two depths and two times during the day. With this approach we attempted to capture the transcriptional diversity associated with change in light and other depth-related environmental factors. Using this extensive data set we generated gene predictions and identified an extensive catalogue of potential Simple Sequence Repeats (SSR) markers. The data generated here will open new avenues for the analysis of population genetic features and functional variation in P. oceanica. In total, 79,235 contigs were obtained by the assembly of 70,453,120 paired end reads. 43,711 contigs were successfully annotated. A total of 17,436 SSR were identified within 13,912 contigs. PMID:27996971

  7. Developing expressed sequence tag libraries and the discovery of simple sequence repeat markers for two species of raspberry (Rubus L.)

    Science.gov (United States)

    Background: Due to a relatively high level of codominant inheritance and transferability within and among taxonomic groups, simple sequence repeat (SSR) markers are important elements in comparative mapping and delineation of genomic regions associated with traits of economic importance. Expressed S...

  8. Leucine-rich repeat-containing G-protein-coupled receptors as markers of adult stem cells

    NARCIS (Netherlands)

    Barker, N.; Clevers, H.

    2010-01-01

    Molecular markers are used to characterize and track adult stem cells. Colon cancer research has led to the identification of 2 related receptors, leucine-rich repeat-containing, G-protein-coupled receptors (Lgr)5 and Lgr6, that are expressed by small populations of cells in a variety of adult organ

  9. Genetic relationship analysis and fingerprint construction of 62 cultivars of Hippeastrum spp.based on ISSR marker%基于ISSR标记的62个朱顶红品种的遗传关系分析及指纹图谱构建

    Institute of Scientific and Technical Information of China (English)

    张林; 徐迎春; 成海钟; 周玉珍; 娄晓鸣; 吕文涛

    2012-01-01

    The genetic diversity of 62 cultivars (including 60 cultivars introduced from Netherlands and 2 local cultivars in Suzhou) of Hippeastrum spp.was analyzed by ISSR marker method, and the cluster analysis on 62 cultivars was carried out by UPGMA method.On the basis, by means of comparison and selection of specific bands, the ISSR fingerprint of tested cultivars was constructed by ways of schematic diagram with black and white square lattices.The amplification result shows that total 118 bands are amplified from genomic DNA of 62 cultivars by 11 primers, in which, there are 109 polymorphic bands with the polymorphic band percentage (PPB) of 92.4% , and PPB of bands amplified by five primers reaches to 100.0% .The change range of genetic similarity coefficient among 62 cultivars is larger with a value from 0.371 4 to 0.842 9, showing that there are rich genetic variation and genetic diversity among different cultivars.The cluster analysis result shows that 62 cultivars can be divided into seven groups at the genetic similarity coefficient of 0.63.Most cultivars with similar morphology are clustered together, in which the genetic similarity coefficient among some similar morphology cultivars with white and single petal is higher (about 0.8) and most of these cultivars are clustered together, indicating that their homology is higher.While, two local cultivars in Suzhou have the highest genetic similarity coefficient (0.842 9) , meaning that the two cultivars may possess a same origin.Cultivar ' Xiaohongxing' has a specific band at 450 bp in amplification pattern of primer UBC873 and cultivar ' Elvas' has a specific missing band at 3 000 bp in amplification pattern of primer UBC835, therefore, these two specific bands can be used for identification of cultivars ' Xiaohongxing' and ' Elvas' , respectively.According to ISSR amplification bands by primers UBC835 and UBC873, the ISSR fingerprint of 62 cultivars is combined and constructed, and all of cultivars tested can be

  10. ISSR标记技术在湖南柑橘主要品种亲缘关系研究中的应用%Application of ISSR Markers in Analysis of Genetic Relationship for Main Varieties of Citrus in Hunan Province

    Institute of Scientific and Technical Information of China (English)

    黄绿红; 刘咏红; 李高阳; 张菊华

    2012-01-01

    利用ISSR分子标记技术对16个湖南柑橘主要品种进行了分子鉴定和亲缘关系分析.结果表明:9个引物共扩增出72条谱带,其中54条为多态性条带,多态率高达75%,平均每条引物扩增的DNA条带数目为8条.·利用Popgene 1.32程序进行了Nei遗传距离和遗传相似性分析,橙类中彭娜和纽荷尔的亲缘关系最近.UPGMA聚类分析结果表明,在遗传距离为0.76的水平上,16个柑橘品种可分为四大类群.%Sixteen main varieties of citrus were used as materials for analyzing their genome polymorphism by ISSR markers. The results showed that total 72 bands were generated by nine primers, of which 54 bands were polymorphic bands (the percentage of polymorphic band, PPB=75%). The average number of DNA fragments produced by each primer was eight. Nei similarity coefficients and genetic distances were calculated by using Popgene 1.32 software, and results indicated that Penna and Newhall have the closest relationship among citrus; the dendrogram were constructed by using UPGMA, and the results indicated that 16 citrus materials were divided into four major groups while the genetic distance was 0.76.

  11. Establishment and Optimization of ISSR-PCR Amplification System in Squash%南瓜 ISSR-PCR 反应体系的建立与优化

    Institute of Scientific and Technical Information of China (English)

    朱海生; 卢丽芳; 温庆放; 林义章

    2014-01-01

    为获得南瓜 ISSR 最优扩增体系,为后续南瓜的遗传多样性分析和亲缘关系鉴定奠定基础,本研究以‘密本’南瓜为筛选体系材料,采用单因素试验,对 ISSR 反应体系的 Mg2+、dNTP、Taq 酶、引物和 DNA 浓度等5个因素进行优化。研究结果表明,南瓜 ISSR 25μL 最佳反应体系为:2.5μL 10×Buffer,2.0 mmol/L Mg2+,0.3 mmol/L dNTP,1 U Taq 酶,0.48 mmol/L 引物,75 ng 的 DNA。在此基础上,对筛选出的12条引物进行退火温度的优化,最终确定每条引物的最佳退火温度。利用该反应体系,选用引物891对85个南瓜品种进行所确立扩增体系的验证,结果显示扩增产物条带清晰明亮、多态性丰富,且特异性强、重复性好,表明本研究所确定的反应体系适用于南瓜的 ISSR 分子标记。%In order to obtain the ISSR-PCR reaction system of Squash, We took an optimization experiment with single factor design which was concentrated on five factors of Mg2+, dNTP, Taq DNA polymerase, primer and template DNA by using the squash ‘miben’as the material of filter system to set up a good foundation of analy-sing the genetic diversity and relationship identification of Squash. The test result showed that the greatest 25 μL ISSR reaction system which contained 2.5 μL 10×Buffer, 2.0 mmol/L Mg2+, 0.3 mmol/L dNTP, 1 U Taq DNA polymerase, 0.48 mmol/L primer, 75 ng template DNA. On this basis, optimized annealing temperature of 12 primers which have been screened, and ultimately determined the optimal annealing temperature of each primer. 85 squashes were amplified with primer 891 under the optimized reaction conditions. The test result showed the amplification products which were clear and bright, abundant polymorphism, strong specificity and good repeatability. Accordingly, it was suitable for ISSR analysis of squash.

  12. Exploring the original causes of cherry cultivar Manaohong originated from Nayong County,Guizhou Province using ISSR markers%利用ISSR标记探讨贵州地方樱桃种质玛瑙红的起源原因

    Institute of Scientific and Technical Information of China (English)

    宋常美; 文晓鹏

    2011-01-01

    In this paper,genetic relationship among local cherry cultivar Manaohong originated from Nayong County of Guizhou Province,as well as:sour cherry,black pearl and four sweet cherry cultivars were investigated using ISSR markers.The purpose was to investigate the causes of variation on Manaohong.A total of 286 markers were scored from 15 primers,which were demonstrated highly reproducible,clear and polymorphic bands.Among the obtained markers,249 were polymorphic,accounting for 87.06% of the total.As analyzed by NTSYS2.01,the genetic similarity among the seven accessions ranged from 0.41 to 0.94,and the genetic similarity between Manaohong and sour cherry was 0.57.Manaohong has significant genetic differences with sour cherry on molecular level,possibly it is a hybridization variation of sour cherry.%为探讨玛瑙红的起源原因,利用ISSR标记对贵州省纳雍县地方樱桃种质玛瑙红、酸樱桃、黑珍珠和4种欧洲甜樱桃等种质的亲缘关系进行分析。结果表明:用15条稳定性强、条带清晰及多态性丰富的引物进行PCR扩增,共获得286个标记位点,其中多态性位点为249个,多态性比例为87.06%;采用NTSYS2.01软件计算,7种樱桃相似性系数为0.41~0.94,玛瑙红与酸樱桃的相似系数为0.57。玛瑙红和酸樱桃在分子水平上的遗传差异很大,该种质可能为酸樱桃的实生变异种质。

  13. Genetic diversity of Cercospora kikuchii isolates from soybean cultured in Argentina as revealed by molecular markers and cercosporin production.

    Science.gov (United States)

    Lurá, María Cristina; Latorre Rapela, María Gabriela; Vaccari, María Celia; Maumary, Roxana; Soldano, Anabel; Mattio, Mónica; González, Ana María

    2011-05-01

    Leaf blight and purple seed, caused by the fungal pathogen Cercospora kikuchii (Matsumoto & Tomoyasu) M. W. Gardner are very important diseases of soybean (Glycine max L. Merr.) in Argentina. The aims of this work were: (a) to confirm and to assess the genetic variability among C. kikuchii isolates collected from different soybean growing areas in Santa Fe province using inter simple sequence repeats (ISSR) markers and sequence information from the internal transcribed spacer (ITS) region of rDNA and (b) to analyze the cercosporin production of the regional C. kikuchi isolates in order to assess whether there was any relationship between the molecular profiles and the toxin production. Isolates from different regions in Santa Fe province were studied. The sequence of the ITS regions showed high similarity (99-100%) to the GenBank sequences of C. kikuchii BRCK179 (accession number AY633838). The ISSR markers clustered all the isolates into many groups and cercosporin content was highly variable among isolates. No relationship was observed between ITS region, ISSR groups and origin or cercosporin content. The high degree of genetic variability and cercosporin production among isolates compared in this study characterizes a diverse population of C. kikuchii in the region.

  14. Genetic Differentiation of Japanese Sardinella (Sardinella zunasi)Populations in the Northwest Pacific Revealed by ISSR Analysis

    Institute of Scientific and Technical Information of China (English)

    YING Yiping; GAO Tianxiang; MIAO Zhenqing

    2011-01-01

    Knowledge of population genetic structure plays an important role in fisheries management.In this research,Inter-Simple-Sequence-Repeat (ISSR) markers were employed to evaluate the genetic structure of Japanese sardinella (Sardinella zunasi) populations in the Northwest Pacific.Eighty seven individuals from 5 locations were screened using 4 highly polymorphic primers.A total of 173 polymorphic loci were detected out of 191 loci amplified.Small but significant genetic differentiation was detected between the Chinese and Japanese populations by both AMOVA and pairwise Fsr analyses,which was further supported by cluster analysis.We consider that climate change during glaciations should be responsible for the genetic differentiation.Isolation by geographic distance among populations was observed,indicating that the distance might also lead to the genetic differentiation.However,no genetic structure was found within the populations off both the Chinese and Japanese coasts,indicating a high-level along-coast gene flow,which might result from ocean current transport and common ground for over-wintering.

  15. ISSR Analysis on Genetic Diversity of the 34 Populations of Oryza meyeriana Distributing in Yunnan Province, China

    Directory of Open Access Journals (Sweden)

    Ya-tao WAN

    2008-03-01

    Full Text Available The genetic diversity of the 34 populations of wild rice Oryza meyeriana Baill. distributed in Yunnan Province, China was analyzed using 13 inter-simple sequence repeat (ISSR markers. A total of 168 bands were amplified, of which 135 polymorphic bands were discovered and the percentage of polymorphic bands (PPB was 80.36%. A genetic diversity was revealed as Nei's gene diversity (H = 0.2666 and Shannon information index (I = 0.4028 at population level. The 34 populations were divided into different groups based on administrative regions, latitude and longitudes, river areas, altitudes of their origins, and their indexes such as Na (number of alleles, Ne (effective number of alleles, H, I and PPB were calculated. Richer genetic diversity was found in the wild rice populations distributed in Simao Prefecture than that in Lingcang Prefecture or Xishuangbanna Prefecture whereas the least genetic diversity was in Baoshan Prefecture or Dehong Prefecture. Rich genetic diversity was also discovered in the wild rice populations originated from higher than 710 m altitude around the middle and lower reaches of the Lancang River belonging to the Pacific Ocean drainage system. The 34 populations could be classified into two groups, one group covered the wild rice distributing in Simao Prefecture only while the other group covered ones in Lingcang, Xishuangbanna and Dehong Prefectures. The issue on how to effectively conserve the wild rice germplasm was discussed.

  16. Genetic relationships among populations of Aedes aegypti from Uruguay and northeastern Argentina inferred from ISSR-PCR data.

    Science.gov (United States)

    Soliani, C; Rondan-Dueñas, J; Chiappero, M B; Martínez, M; Da Rosa, E García; Gardenal, C N

    2010-09-01

    Aedes aegypti (L.) (Diptera: Culicidae), the main vector of yellow fever and dengue viruses, was eradicated from Argentina between 1955 and 1963, but reinvaded the country in 1986. In Uruguay, the species was reintroduced in 1997. In this study we used highly polymorphic inter-simple sequence repeats (ISSR) markers to analyse the genetic structure of Ae. aegypti populations from Uruguay and northeastern Argentina to identify possible colonization patterns of the vector. Overall genetic differentiation among populations was high (F(ST) = 0.106) and showed no correlation with geographic distance, which is consistent with the short time since the reintroduction of the species in the area. Differentiation between pairs of Argentine populations (F(ST) 0.072 to 0.221) was on average higher than between Uruguayan populations (F(ST)-0.044 to 0.116). Bayesian estimation of population structure defined four genetic clusters and most populations were admixtures of two of them: Mercedes and Treinta y Tres (Uruguay) were mixtures of clusters 1 and 3; Salto (Uruguay) and Paraná (Argentina) of clusters 1 and 4; Fray Bentos (Uruguay) of clusters 2 and 3, and Gualeguaychú (Argentina) of clusters 2 and 3. Posadas and Buenos Aires in Argentina were fairly genetically homogeneous. Our results suggest that Ae. aegypti recolonized Uruguay from bordering cities in Argentina via bridges over the Uruguay River and also from Brazil.

  17. 菠萝蜜遗传多样性的ISSR分析%Analysis of genetic diversity of Jackfruit germplasm using ISSR marking method

    Institute of Scientific and Technical Information of China (English)

    叶春海; 王耀辉; 李映志; 丰锋

    2009-01-01

    ISSR (inter-simple sequence repeat)markers were used to analyze genetic diversity in 76 accessions of jackfruit (Artocarpus heterophyllus Lain.). Among 477 bands produced by 24 ISSR primers, 427 were polymorphic (accounted for 89.52%). The average PIC (pelymorphism information content) was 0.23. These 76 jackfruit accessions could be discriminated by the 24 ISSR primers. Genetic distance analysis revealed that, low genetic diversity existed in the jackfruit germplasm studied, and the genetic similarity coefficient ranged from 0.626 to 0.945, with an average of 0.775. Cluster analysis showed that 4 groups could be clustered at a genetic distance coefficient of 0.752. The results indicated that, 2 accessions from Tropical Plant Garden, WJ2 and GSYWJ1, were genetically different from other accessions. Accessions of soft flesh type and firm flesh type could not be discriminated. Accessions collected from different sources could form independent clusters, while accessions collected from different areas of Leizhou peninsula could not form independent clusters.%用ISSR标记方法对76份菠萝蜜(Artocarpus heterophyllus Lam.)种质资源DNA的遗传多样性进行了检测.24个引物共检测到477条带,其中427条具多态性(占89052%),平均PIC为0.23,24个引物能把76份种质完全区分开来.遗传距离分析结果显示供试种质的遗传多样性较低,在DNA水平上的遗传相似系数为0.626~0.945,平均为0.775.聚类分析表明,76份菠萝蜜材料在遗传距离系数为0.752处可分为4大类,其中热带植物园的种质WJ2和GSYWJ1与其他种质的遗传距离相对较大,干胞和湿类型不能独立聚类.另外各地区的种质与雷州半岛种质明显分开聚类,而雷州半岛各地区的种质混杂聚类在一起,不能按地区单独聚类.

  18. The Comparison of ISSR and PAPD Analysis of Edible Wild Mushrooms of Anhui%安徽野生香菇ISSR和PAPD分析的比较

    Institute of Scientific and Technical Information of China (English)

    王子迎

    2005-01-01

    采用随机扩增多态性DNA(RAPD)和简单序列重复区间扩增多态性(Inter-simple sequence repeat,ISSR)技术,利用12条RAPD引物和10条ISSR引物对10个安徽香菇菌株进行了DNA标记比较分析.结果表明,ISSR-PCR较RAPD-PCR稳定,更具可操作性;供试菌株存在比较显著的遗传变异,具有较丰富的遗传多样性.

  19. Diversity Analysis in Cannabis sativa Based on Large-Scale Development of Expressed Sequence Tag-Derived Simple Sequence Repeat Markers

    OpenAIRE

    Chunsheng Gao; Pengfei Xin; Chaohua Cheng; Qing Tang; Ping Chen; Changbiao Wang; Gonggu Zang; Lining Zhao

    2014-01-01

    Cannabis sativa L. is an important economic plant for the production of food, fiber, oils, and intoxicants. However, lack of sufficient simple sequence repeat (SSR) markers has limited the development of cannabis genetic research. Here, large-scale development of expressed sequence tag simple sequence repeat (EST-SSR) markers was performed to obtain more informative genetic markers, and to assess genetic diversity in cannabis (Cannabis sativa L.). Based on the cannabis transcriptome, 4,577 SS...

  20. ISSR-PCR技术在植物种间和种下关系中的应用%Application of ISSR - PCR technology on the relation of interspecific and under specific

    Institute of Scientific and Technical Information of China (English)

    邓辉; 黄晓燕; 乙引

    2006-01-01

    ISSR-PCR(inter-simple sequence repeat-PCR)标记技术是在SSR-PCR基础上发展起来的一项新的DNA分子标记技术,具有操作简便、重复性好、多态性高等特点.本文对ISSR技术及其在植物种间和种下关系中的应用进行了概述.

  1. Exploiting BAC-end sequences for the mining, characterization and utility of new short sequences repeat (SSR) markers in Citrus.

    Science.gov (United States)

    Biswas, Manosh Kumar; Chai, Lijun; Mayer, Christoph; Xu, Qiang; Guo, Wenwu; Deng, Xiuxin

    2012-05-01

    The aim of this study was to develop a large set of microsatellite markers based on publicly available BAC-end sequences (BESs), and to evaluate their transferability, discriminating capacity of genotypes and mapping ability in Citrus. A set of 1,281 simple sequence repeat (SSR) markers were developed from the 46,339 Citrus clementina BAC-end sequences (BES), of them 20.67% contained SSR longer than 20 bp, corresponding to roughly one perfect SSR per 2.04 kb. The most abundant motifs were di-nucleotide (16.82%) repeats. Among all repeat motifs (TA/AT)n is the most abundant (8.38%), followed by (AG/CT)n (4.51%). Most of the BES-SSR are located in the non-coding region, but 1.3% of BES-SSRs were found to be associated with transposable element (TE). A total of 400 novel SSR primer pairs were synthesized and their transferability and polymorphism tested on a set of 16 Citrus and Citrus relative's species. Among these 333 (83.25%) were successfully amplified and 260 (65.00%) showed cross-species transferability with Poncirus trifoliata and Fortunella sp. These cross-species transferable markers could be useful for cultivar identification, for genomic study of Citrus, Poncirus and Fortunella sp. Utility of the developed SSR marker was demonstrated by identifying a set of 118 markers each for construction of linkage map of Citrus reticulata and Poncirus trifoliata. Genetic diversity and phylogenetic relationship among 40 Citrus and its related species were conducted with the aid of 25 randomly selected SSR primer pairs and results revealed that citrus genomic SSRs are superior to genic SSR for genetic diversity and germplasm characterization of Citrus spp.

  2. 二倍体与四倍体苹果的三倍性杂交后代 ISSR 遗传分析%Genetic Analysis of Apple Triploid Hybrid Progenies from Diploid and Tetraploid Based on ISSR

    Institute of Scientific and Technical Information of China (English)

    何平; 王玉霞; 李林光; 王海波; 李慧峰; 孙清荣; 程来亮

    2015-01-01

    利用 ISSR 分子标记技术对苹果‘寒富’与‘四倍体嘎拉’杂交的三倍性后代及亲本进行遗传变异研究。结果表明9条 ISSR 引物共扩增出多态性条带58条,多态性比率为67.4%。相似性分析可知,三倍性新种质与母体相似系数在0.5909~0.7692之间,与父本相似系数在0.5714~0.7500之间,聚类结果显示三倍性杂交后代具有母本倾向。%The genetic variation of triploid hybrid progenies from ‘Hanfu’ב4 x Gala’were studied based on ISSR molecular markers in this paper.The results showed that 58 polymorphic bands were amplified by 9 ISSR primers,the percentage of polymorphic bands of ISSR was 67.4%.The genetic similarity was 0.5909 ~0.7692 between triploid hybrid progenies and female parent,and it was 0.5714 ~0.7500 between triploid hybrid progenies and male parent.The cluster result showed that most of triploid hybrid progenies were tend to the female parent ‘Hanfu’.

  3. Expressed Sequence Tag-Simple Sequence Repeat (EST-SSR Marker Resources for Diversity Analysis of Mango (Mangifera indica L.

    Directory of Open Access Journals (Sweden)

    Natalie L. Dillon

    2014-01-01

    Full Text Available In this study, a collection of 24,840 expressed sequence tags (ESTs generated from five mango (Mangifera indica L. cDNA libraries was mined for EST-based simple sequence repeat (SSR markers. Over 1,000 ESTs with SSR motifs were detected from more than 24,000 EST sequences with di- and tri-nucleotide repeat motifs the most abundant. Of these, 25 EST-SSRs in genes involved in plant development, stress response, and fruit color and flavor development pathways were selected, developed into PCR markers and characterized in a population of 32 mango selections including M. indica varieties, and related Mangifera species. Twenty-four of the 25 EST-SSR markers exhibited polymorphisms, identifying a total of 86 alleles with an average of 5.38 alleles per locus, and distinguished between all Mangifera selections. Private alleles were identified for Mangifera species. These newly developed EST-SSR markers enhance the current 11 SSR mango genetic identity panel utilized by the Australian Mango Breeding Program. The current panel has been used to identify progeny and parents for selection and the application of this extended panel will further improve and help to design mango hybridization strategies for increased breeding efficiency.

  4. ISSR Fingerprint for Manchurian Tigers in Captivity%圈养东北虎ISSR指纹分析初报

    Institute of Scientific and Technical Information of China (English)

    白秀娟

    2004-01-01

    Manchurian tigers kept in Hengdaohezi Breeding Center,Heilongjiang province,were fingerprinted using inter simple sequence repeated (ISSR) technique.Seven primers were screened out of 20 primers with 41 sites deprived,The results indicated that 31 bands were polymorphie sites and the frequency was 76%.The maximum genetic similarity coefficients among the 15 tigers were 0.526 3 and 0.0909,respectively.The average genetic similarity coefficient of the population was 0.327 1.

  5. Inter Simple Sequence Repeat (ISSR) analysis of wild and cultivated ...

    African Journals Online (AJOL)

    ONOS

    2010-08-09

    Aug 9, 2010 ... based on populations of origin. Oryza glaberrima ... cultivated species will enhance the genetic variation essential to rice .... two algorithms is that UPGMA assumes equal rates of evolution. (molecular clock ..... distinguished at the phenotypic level from native cultivar ..... and Development series. Doctoral ...

  6. Association Analysis of Simple Sequence Repeat (SSR Markers with Agronomic Traits in Tall Fescue (Festuca arundinacea Schreb..

    Directory of Open Access Journals (Sweden)

    Yanhong Lou

    Full Text Available Tall fescue is widely used in temperate regions throughout the world as a dominant forage grass as well as a turfgrass, in pastoral and turf industry. However, the utilization of tall fescue was limited because of its leaf roughness, poor regeneration ability and poor stress resistance. New cultivars were desirable in modern pastoral industries exceed the potential of existing cultivars. Therefore, well understanding the agronomic traits and describing germplasms would help to overcome these constraints, and morphological evaluation of tall fescue germplasm is the key component in selecting rational parents for hybridization breeding. However, describing the morphological traits of tall fescue germplasm is costly and time-consuming. Fortunately, biotechnology approaches can supplement conventional breeding efforts for tall fescue improvement. Association mapping, as a powerful approach to identify association between agronomic traits and molecular markers has been widely used for enhancing the utilization, conservation and management of the tall fescue germplasms. Therefore, in the present research, 115 tall fescue accessions from different origins (25 accessions are cultivars; 31 accessions from America; 32 accessions from European; 7 accessions from Africa; 20 accessions from Asia, were evaluated for agronomic traits and genetic diversity with 90 simple sequence repeat (SSR markers. The panel displayed significant variation in spike count per plant (SCP and spike weight (SW. However, BCS performed the lowest CV among all the observed agronomic traits. Three subpopulations were identified within the collections but no obvious relative kinship (K was found. The GLM model was used to describe the association between SSR and agronomic traits. Fifty-one SSR markers associated with agronomic traits were observed. Twelve single-associated markers were associated with PH; six single-associated markers were associated with BCS; eight single

  7. Molecular markers for population genetic analyses in the family Psittacidae (Psittaciformes, Aves

    Directory of Open Access Journals (Sweden)

    Patrícia J. Faria

    2006-01-01

    Full Text Available The selection of molecular markers for population studies is an important tool for biodiversity conservation. The family Psittacidae contains many endangered and vulnerable species and we tested three kinds of molecular markers for their potential use in population studies of five psitacid species: 43 hyacinth macaws (Anodorhynchus hyacinthinus, 42 blue-and-yellow macaws (Ara ararauna, 23 red-and-green macaws (Ara chloroptera, 19 red-spectacled amazons (Amazona pretrei; and 18 red-tailed amazons (Amazona brasiliensis. We tested 21 clones from a genomic library of golden conure (Guarouba guarouba minisatellites and 12 pairs of microsatellite primers developed for the domestic chicken (Gallus gallus and A. hyacinthinus. We also tested seven tetranucleotide repeat primers for their ability to amplify regions between microsatellite loci (inter simple sequence repeats, ISSRs. We were able to select seven markers that were variable in different degrees for three species (A. hyacinthinus, A. chloroptera and A. ararauna. The mini and microsatellites produced more polymorphic patterns than the ISSRs. The genetic variability of the species studied seems to be correlated with their endangered status.

  8. Survey and analysis of simple sequence repeats in the Laccaria bicolor genome, with development of microsatellite markers

    Energy Technology Data Exchange (ETDEWEB)

    Labbe, Jessy L [ORNL; Murat, Claude [INRA, Nancy, France; Morin, Emmanuelle [INRA, Nancy, France; Le Tacon, F [UMR, France; Martin, Francis [INRA, Nancy, France

    2011-01-01

    It is becoming clear that simple sequence repeats (SSRs) play a significant role in fungal genome organization, and they are a large source of genetic markers for population genetics and meiotic maps. We identified SSRs in the Laccaria bicolor genome by in silico survey and analyzed their distribution in the different genomic regions. We also compared the abundance and distribution of SSRs in L. bicolor with those of the following fungal genomes: Phanerochaete chrysosporium, Coprinopsis cinerea, Ustilago maydis, Cryptococcus neoformans, Aspergillus nidulans, Magnaporthe grisea, Neurospora crassa and Saccharomyces cerevisiae. Using the MISA computer program, we detected 277,062 SSRs in the L. bicolor genome representing 8% of the assembled genomic sequence. Among the analyzed basidiomycetes, L. bicolor exhibited the highest SSR density although no correlation between relative abundance and the genome sizes was observed. In most genomes the short motifs (mono- to trinucleotides) were more abundant than the longer repeated SSRs. Generally, in each organism, the occurrence, relative abundance, and relative density of SSRs decreased as the repeat unit increased. Furthermore, each organism had its own common and longest SSRs. In the L. bicolor genome, most of the SSRs were located in intergenic regions (73.3%) and the highest SSR density was observed in transposable elements (TEs; 6,706 SSRs/Mb). However, 81% of the protein-coding genes contained SSRs in their exons, suggesting that SSR polymorphism may alter gene phenotypes. Within a L. bicolor offspring, sequence polymorphism of 78 SSRs was mainly detected in non-TE intergenic regions. Unlike previously developed microsatellite markers, these new ones are spread throughout the genome; these markers could have immediate applications in population genetics.

  9. Development and Characterization of Simple Sequence Repeat Markers Providing Genome-Wide Coverage and High Resolution in Maize

    Science.gov (United States)

    Xu, Jie; Liu, Ling; Xu, Yunbi; Chen, Churun; Rong, Tingzhao; Ali, Farhan; Zhou, Shufeng; Wu, Fengkai; Liu, Yaxi; Wang, Jing; Cao, Moju; Lu, Yanli

    2013-01-01

    Simple sequence repeats (SSRs) have been widely used in maize genetics and breeding, because they are co-dominant, easy to score, and highly abundant. In this study, we used whole-genome sequences from 16 maize inbreds and 1 wild relative to determine SSR abundance and to develop a set of high-density polymorphic SSR markers. A total of 264 658 SSRs were identified across the 17 genomes, with an average of 135 693 SSRs per genome. Marker density was one SSR every of 15.48 kb. (C/G)n, (AT)n, (CAG/CTG)n, and (AAAT/ATTT)n were the most frequent motifs for mono, di-, tri-, and tetra-nucleotide SSRs, respectively. SSRs were most abundant in intergenic region and least frequent in untranslated regions, as revealed by comparing SSR distributions of three representative resequenced genomes. Comparing SSR sequences and e-polymerase chain reaction analysis among the 17 tested genomes created a new database, including 111 887 SSRs, that could be develop as polymorphic markers in silico. Among these markers, 58.00, 26.09, 7.20, 3.00, 3.93, and 1.78% of them had mono, di-, tri-, tetra-, penta-, and hexa-nucleotide motifs, respectively. Polymorphic information content for 35 573 polymorphic SSRs out of 111 887 loci varied from 0.05 to 0.83, with an average of 0.31 in the 17 tested genomes. Experimental validation of polymorphic SSR markers showed that over 70% of the primer pairs could generate the target bands with length polymorphism, and these markers would be very powerful when they are used for genetic populations derived from various types of maize germplasms that were sampled for this study. PMID:23804557

  10. The First Molecular Identification of an Olive Collection Applying Standard Simple Sequence Repeats and Novel Expressed Sequence Tag Markers

    Directory of Open Access Journals (Sweden)

    Soraya Mousavi

    2017-07-01

    Full Text Available Germplasm collections of tree crop species represent fundamental tools for conservation of diversity and key steps for its characterization and evaluation. For the olive tree, several collections were created all over the world, but only few of them have been fully characterized and molecularly identified. The olive collection of Perugia University (UNIPG, established in the years’ 60, represents one of the first attempts to gather and safeguard olive diversity, keeping together cultivars from different countries. In the present study, a set of 370 olive trees previously uncharacterized was screened with 10 standard simple sequence repeats (SSRs and nine new EST-SSR markers, to correctly and thoroughly identify all genotypes, verify their representativeness of the entire cultivated olive variation, and validate the effectiveness of new markers in comparison to standard genotyping tools. The SSR analysis revealed the presence of 59 genotypes, corresponding to 72 well known cultivars, 13 of them resulting exclusively present in this collection. The new EST-SSRs have shown values of diversity parameters quite similar to those of best standard SSRs. When compared to hundreds of Mediterranean cultivars, the UNIPG olive accessions were splitted into the three main populations (East, Center and West Mediterranean, confirming that the collection has a good representativeness of the entire olive variability. Furthermore, Bayesian analysis, performed on the 59 genotypes of the collection by the use of both sets of markers, have demonstrated their splitting into four clusters, with a well balanced membership obtained by EST respect to standard SSRs. The new OLEST (Olea expressed sequence tags SSR markers resulted as effective as the best standard markers. The information obtained from this study represents a high valuable tool for ex situ conservation and management of olive genetic resources, useful to build a common database from worldwide olive

  11. A six degrees-of-freedom marker set for gait analysis: repeatability and comparison with a modified Helen Hayes set.

    Science.gov (United States)

    Collins, Thomas D; Ghoussayni, Salim N; Ewins, David J; Kent, Jenny A

    2009-08-01

    Kinematic gait analysis is limited by simplified marker sets and related models. The majority of sets in clinical use were developed with low resolution imaging systems so required various assumptions about body behaviour. Further major limitations include soft tissue artefact and ambiguity in landmark identification. An alternative is the use of sets based on six degrees-of-freedom (DOF) principles, primarily using marker clusters for tracking. This study evaluates performance of a 6DOF set, based largely on CAST/ISB recommendations, through comparison with a conventional set and assessment of repeatability. Ten healthy subjects were assessed in treadmill walking, with both sets applied simultaneously on two occasions. Data were analysed using repeatability coefficients, correlation of key features, and comparison of joint angle curves and difference curves with confidence bands. Apart from pelvic tilt all segment and joint angles from both sets showed high within and between session repeatability (CMC>0.80). Hip rotations showed clear differences between the two sets with indications in support of the 6DOF set. Knee coronal angles showed evidence of cross-talk in the conventional set, highlighting difficulties with anatomical identification despite control measures such as a foot alignment template. Knee transverse angles showed inconsistent patterns for both sets. At the ankle the conventional set only allowed true measurement in two planes so with high repeatability the 6DOF set is preferable. The 6DOF set showed comparable performance to the conventional set and overcomes a number of theoretical limitations, however further development is needed prior to clinical implementation.

  12. Selecting soybean resistant to the cyst nematode Heterodera glycines using simple sequence repeat (microssatellite) markers.

    Science.gov (United States)

    Espindola, S M C G; Hamawaki, O T; Oliveira, A P; Hamawaki, C D L; Hamawaki, R L; Takahashi, L M

    2016-03-11

    The soybean cyst nematode (SCN) is a major cause of soybean yield reduction. The objective of this study was to evaluate the efficiency of marker-assisted selection to identify genotypes resistant to SCN race 3 infection, using Sat_168 and Sat-141 resistance quantitative trait loci. The experiment was carried out under greenhouse conditions, using soybean populations originated from crosses between susceptible and resistant parent stock: CD-201 (susceptible) and Foster IAC (resistant), Conquista (susceptible) and S83-30 (resistant), La-Suprema (susceptible) and S57-11 (resistant), and Parecis (susceptible) and S65-50 (resistant). Plants were inoculated with SCN and evaluated according to the female index (FI), those with FI < 10% were classified as resistant to nematode infection. Plants were genotyped for SCN resistance using microsatellite markers Sat-141 and Sat_168. Marker selection efficiency was analyzed by a contingency table, taking into account genotypic versus phenotypic evaluations for each line. These markers were shown to be useful tool for selection of SCN race 3.

  13. Survey and analysis of simple sequence repeats in the Ustilaginoidea virens genome and the development of microsatellite markers.

    Science.gov (United States)

    Yu, Mina; Yu, Junjie; Li, Huanhuan; Wang, Yahui; Yin, Xiaole; Bo, Huiwen; Ding, Hui; Zhou, Yuxin; Liu, Yongfeng

    2016-07-01

    Ustilaginoidea virens is the causal agent of rice false smut, causing quantitative and qualitative losses in rice industry. However, the development and application of simple sequence repeat (SSR) markers for genetic diversity studies in U. virens were limited. This study is the first to perform large-scale development of SSR markers of this pathogen at the genome level, to (1) compare these SSR markers with those of other fungi, (2) analyze the pattern of the SSRs, and (3) obtain more informative genetic markers. U. virens is rich in SSRs, and 13,778 SSRs were identified with a relative abundance of 349.7SSRs/Mb. The most common motifs in the genome or in noncoding regions were mononucleotides, whereas trinucleotides in coding sequences. A total of 6 out of 127 primers were randomly selected to be used to analyze 115 isolates, and these 6 primers showed high polymorphism in U. virens. This study may serve as an important resource for molecular genetic studies in U. virens.

  14. Genetic and phytochemical analysis of the in vitro regenerated Pilosocereus robinii by ISSR, SDS-PAGE and HPLC.

    Science.gov (United States)

    Khattab, Salah; El Sherif, Fadia; El-Garhy, Hoda A; Ahmed, Safwat; Ibrahim, Amany

    2014-01-01

    Pilosocereus robinii is a rare species which is experiencing sudden population collapse. Identifying and developing effective conservation and management strategies to halt the forestall extinction of this species is crucial. The present study was conducted to assess the best conditions for in vitro propagation of this plant in regard to its morphogenic, genetic as well as the chemical potentials. A successful in vitro propagation system of P. robinii has been developed. MS hormone-free medium induced the best root morphogenic potential. The plants were acclimatized in the greenhouse at 100% survival rate. Besides, the somaclonal variations between the in vitro raised plants were analyzed using PCR-ISSR markers and SDS-PAGE protein, where the regenerated explants on MS medium supplemented with TDZ were the highest in inducing new specific marker bands. Sh6 ISSR primer showed the highest polymorphism value, 81.8% with 33 total amplified fragments, while Sh3 ISSR primer showed the lowest value with polymorphic percentage of 14.3%. Furthermore, SDS-PAGE protein analysis showed no variation in protein pattern of the studied treatments. On the other side, HPLC analysis of the in vitro plantlets extracts has shown that 2iP based treatments were the highest in organic acids accumulation, while the phenolic constituents' accumulation was found to reach its peak in the BA based treatments.

  15. PCR bias in amplification of androgen receptor alleles, a trinucleotide repeat marker used in clonality studies.

    OpenAIRE

    Mutter, G L; Boynton, K A

    1995-01-01

    Trinucleotide CAG repeats in the X-linked human androgen receptor gene (HUMARA) have proved a useful means of determining X chromosome haplotypes, and when combined with methylation analysis of nearby cytosine residues permits identification of non-random X inactivation in tumors of women. Co-amplification of two alleles in a heterozygote generates PCR products which differ in the number of CAG units, and thus their melting and secondary structure characteristics. We have shown that under opt...

  16. Analysis of simple sequence repeats in the Gaeumannomyces graminis var. tritici genome and the development of microsatellite markers.

    Science.gov (United States)

    Li, Wei; Feng, Yanxia; Sun, Haiyan; Deng, Yuanyu; Yu, Hanshou; Chen, Huaigu

    2014-11-01

    Understanding the genetic structure of Gaeumannomyces graminis var. tritici is essential for the establishment of efficient disease control strategies. It is becoming clear that microsatellites, or simple sequence repeats (SSRs), play an important role in genome organization and phenotypic diversity, and are a large source of genetic markers for population genetics and meiotic maps. In this study, we examined the G. graminis var. tritici genome (1) to analyze its pattern of SSRs, (2) to compare it with other plant pathogenic filamentous fungi, such as Magnaporthe oryzae and M. poae, and (3) to identify new polymorphic SSR markers for genetic diversity. The G. graminis var. tritici genome was rich in SSRs; a total 13,650 SSRs have been identified with mononucleotides being the most common motifs. In coding regions, the densities of tri- and hexanucleotides were significantly higher than in noncoding regions. The di-, tri-, tetra, penta, and hexanucleotide repeats in the G. graminis var. tritici genome were more abundant than the same repeats in M. oryzae and M. poae. From 115 devised primers, 39 SSRs are polymorphic with G. graminis var. tritici isolates, and 8 primers were randomly selected to analyze 116 isolates from China. The number of alleles varied from 2 to 7 and the expected heterozygosity (He) from 0.499 to 0.837. In conclusion, SSRs developed in this study were highly polymorphic, and our analysis indicated that G. graminis var. tritici is a species with high genetic diversity. The results provide a pioneering report for several applications, such as the assessment of population structure and genetic diversity of G. graminis var. tritici.

  17. Development of expressed sequence tag and expressed sequence tag–simple sequence repeat marker resources for Musa acuminata

    Science.gov (United States)

    Passos, Marco A. N.; de Oliveira Cruz, Viviane; Emediato, Flavia L.; de Camargo Teixeira, Cristiane; Souza, Manoel T.; Matsumoto, Takashi; Rennó Azevedo, Vânia C.; Ferreira, Claudia F.; Amorim, Edson P.; de Alencar Figueiredo, Lucio Flavio; Martins, Natalia F.; de Jesus Barbosa Cavalcante, Maria; Baurens, Franc-Christophe; da Silva, Orzenil Bonfim; Pappas, Georgios J.; Pignolet, Luc; Abadie, Catherine; Ciampi, Ana Y.; Piffanelli, Pietro; Miller, Robert N. G.

    2012-01-01

    Background and aims Banana (Musa acuminata) is a crop contributing to global food security. Many varieties lack resistance to biotic stresses, due to sterility and narrow genetic background. The objective of this study was to develop an expressed sequence tag (EST) database of transcripts expressed during compatible and incompatible banana–Mycosphaerella fijiensis (Mf) interactions. Black leaf streak disease (BLSD), caused by Mf, is a destructive disease of banana. Microsatellite markers were developed as a resource for crop improvement. Methodology cDNA libraries were constructed from in vitro-infected leaves from BLSD-resistant M. acuminata ssp. burmaniccoides Calcutta 4 (MAC4) and susceptible M. acuminata cv. Cavendish Grande Naine (MACV). Clones were 5′-end Sanger sequenced, ESTs assembled with TGICL and unigenes annotated using BLAST, Blast2GO and InterProScan. Mreps was used to screen for simple sequence repeats (SSRs), with markers evaluated for polymorphism using 20 diploid (AA) M. acuminata accessions contrasting in resistance to Mycosphaerella leaf spot diseases. Principal results A total of 9333 high-quality ESTs were obtained for MAC4 and 3964 for MACV, which assembled into 3995 unigenes. Of these, 2592 displayed homology to genes encoding proteins with known or putative function, and 266 to genes encoding proteins with unknown function. Gene ontology (GO) classification identified 543 GO terms, 2300 unigenes were assigned to EuKaryotic orthologous group categories and 312 mapped to Kyoto Encyclopedia of Genes and Genomes pathways. A total of 624 SSR loci were identified, with trinucleotide repeat motifs the most abundant in MAC4 (54.1 %) and MACV (57.6 %). Polymorphism across M. acuminata accessions was observed with 75 markers. Alleles per polymorphic locus ranged from 2 to 8, totalling 289. The polymorphism information content ranged from 0.08 to 0.81. Conclusions This EST collection offers a resource for studying functional genes, including

  18. Cytogenetic analysis of Populus trichocarpa--ribosomal DNA, telomere repeat sequence, and marker-selected BACs.

    Science.gov (United States)

    Islam-Faridi, M N; Nelson, C D; DiFazio, S P; Gunter, L E; Tuskan, G A

    2009-01-01

    The 18S-28S rDNA and 5S rDNA loci in Populus trichocarpa were localized using fluorescent in situ hybridization (FISH). Two 18S-28S rDNA sites and one 5S rDNA site were identified and located at the ends of 3 different chromosomes. FISH signals from the Arabidopsis-type telomere repeat sequence were observed at the distal ends of each chromosome. Six BAC clones selected from 2 linkage groups based on genome sequence assembly (LG-I and LG-VI) were localized on 2 chromosomes, as expected. BACs from LG-I hybridized to the longest chromosome in the complement. All BAC positions were found to be concordant with sequence assembly positions. BAC-FISH will be useful for delineating each of the Populus trichocarpa chromosomes and improving the sequence assembly of this model angiosperm tree species.

  19. Cytogenetic Analysis of Populus trichocarpa - Ribosomal DNA, Telomere Repeat Sequence, and Marker-selected BACs

    Energy Technology Data Exchange (ETDEWEB)

    Tuskan, Gerald A [ORNL; Gunter, Lee E [ORNL; DiFazio, Stephen P [West Virginia University

    2009-01-01

    The 18S-28S rDNA and 5S rDNA loci in Populus trichocarpa were localized using fluorescent in situ hybridization (FISH). Two 18S-28S rDNA sites and one 5S rDNA site were identified and located at the ends of 3 different chromosomes. FISH signals from the Arabidopsis -type telomere repeat sequence were observed at the distal ends of each chromosome. Six BAC clones selected from 2 linkage groups based on genome sequence assembly (LG-I and LG-VI) were localized on 2 chromosomes, as expected. BACs from LG-I hybridized to the longest chromosome in the complement. All BAC positions were found to be concordant with sequence assembly positions. BAC-FISH will be useful for delineating each of the Populus trichocarpa chromosomes and improving the sequence assembly of this model angiosperm tree species.

  20. Diversity analysis in Cannabis sativa based on large-scale development of expressed sequence tag-derived simple sequence repeat markers.

    Science.gov (United States)

    Gao, Chunsheng; Xin, Pengfei; Cheng, Chaohua; Tang, Qing; Chen, Ping; Wang, Changbiao; Zang, Gonggu; Zhao, Lining

    2014-01-01

    Cannabis sativa L. is an important economic plant for the production of food, fiber, oils, and intoxicants. However, lack of sufficient simple sequence repeat (SSR) markers has limited the development of cannabis genetic research. Here, large-scale development of expressed sequence tag simple sequence repeat (EST-SSR) markers was performed to obtain more informative genetic markers, and to assess genetic diversity in cannabis (Cannabis sativa L.). Based on the cannabis transcriptome, 4,577 SSRs were identified from 3,624 ESTs. From there, a total of 3,442 complementary primer pairs were designed as SSR markers. Among these markers, trinucleotide repeat motifs (50.99%) were the most abundant, followed by hexanucleotide (25.13%), dinucleotide (16.34%), tetranucloetide (3.8%), and pentanucleotide (3.74%) repeat motifs, respectively. The AAG/CTT trinucleotide repeat (17.96%) was the most abundant motif detected in the SSRs. One hundred and seventeen EST-SSR markers were randomly selected to evaluate primer quality in 24 cannabis varieties. Among these 117 markers, 108 (92.31%) were successfully amplified and 87 (74.36%) were polymorphic. Forty-five polymorphic primer pairs were selected to evaluate genetic diversity and relatedness among the 115 cannabis genotypes. The results showed that 115 varieties could be divided into 4 groups primarily based on geography: Northern China, Europe, Central China, and Southern China. Moreover, the coefficient of similarity when comparing cannabis from Northern China with the European group cannabis was higher than that when comparing with cannabis from the other two groups, owing to a similar climate. This study outlines the first large-scale development of SSR markers for cannabis. These data may serve as a foundation for the development of genetic linkage, quantitative trait loci mapping, and marker-assisted breeding of cannabis.

  1. Diversity analysis in Cannabis sativa based on large-scale development of expressed sequence tag-derived simple sequence repeat markers.

    Directory of Open Access Journals (Sweden)

    Chunsheng Gao

    Full Text Available Cannabis sativa L. is an important economic plant for the production of food, fiber, oils, and intoxicants. However, lack of sufficient simple sequence repeat (SSR markers has limited the development of cannabis genetic research. Here, large-scale development of expressed sequence tag simple sequence repeat (EST-SSR markers was performed to obtain more informative genetic markers, and to assess genetic diversity in cannabis (Cannabis sativa L.. Based on the cannabis transcriptome, 4,577 SSRs were identified from 3,624 ESTs. From there, a total of 3,442 complementary primer pairs were designed as SSR markers. Among these markers, trinucleotide repeat motifs (50.99% were the most abundant, followed by hexanucleotide (25.13%, dinucleotide (16.34%, tetranucloetide (3.8%, and pentanucleotide (3.74% repeat motifs, respectively. The AAG/CTT trinucleotide repeat (17.96% was the most abundant motif detected in the SSRs. One hundred and seventeen EST-SSR markers were randomly selected to evaluate primer quality in 24 cannabis varieties. Among these 117 markers, 108 (92.31% were successfully amplified and 87 (74.36% were polymorphic. Forty-five polymorphic primer pairs were selected to evaluate genetic diversity and relatedness among the 115 cannabis genotypes. The results showed that 115 varieties could be divided into 4 groups primarily based on geography: Northern China, Europe, Central China, and Southern China. Moreover, the coefficient of similarity when comparing cannabis from Northern China with the European group cannabis was higher than that when comparing with cannabis from the other two groups, owing to a similar climate. This study outlines the first large-scale development of SSR markers for cannabis. These data may serve as a foundation for the development of genetic linkage, quantitative trait loci mapping, and marker-assisted breeding of cannabis.

  2. Standardization of DNA isolation and PCR protocols for ISSR analysis in Cluster bean [Cyamopsis tetragonoloba (L. Taub.] Genotypes

    Directory of Open Access Journals (Sweden)

    Vikas Kumar,

    2015-05-01

    Full Text Available Cluster bean [Cyamopsis tetragonoloba (L Taub.] is one of the most important cash legume crop has played an increasingly important role in wide range of industries. Owing to the significance of molecular marker studies in numerous applications including in genetic improvement of crops, there is an obvious need to undertake such studies in cluster bean. Present research work was carried out with the genomic DNA optimization and PCR standardization for ISSR analysis in cluster bean genotypes. The standard CTAB method for isolation of genomic DNA from cluster bean have their own limitation as it produces gel image with high contamination of yellowish, sticky and viscous matrix. The commonly used CTAB method was modified by giving Polyvinylpyrrolidone (PVP treatments after grinding fresh young leaves of cluster bean. The yield of genomic DNA was increased in ranged from120 ng/µl to 220 ng/µl of 0.2 g leaf tissue and the purity (ratio was between 1.5-1.7 indicating minimal levels of contaminating metabolites. ISSR protocol was standardized after implementing various modifications. Based on above findings of present experiment work this modified genomic DNA isolation and PCR protocol may be utilized for ISSR profiling to produce sharp and clear bands in cluster bean genotypes.

  3. PCR bias in amplification of androgen receptor alleles, a trinucleotide repeat marker used in clonality studies.

    Science.gov (United States)

    Mutter, G L; Boynton, K A

    1995-04-25

    Trinucleotide CAG repeats in the X-linked human androgen receptor gene (HUMARA) have proved a useful means of determining X chromosome haplotypes, and when combined with methylation analysis of nearby cytosine residues permits identification of non-random X inactivation in tumors of women. Co-amplification of two alleles in a heterozygote generates PCR products which differ in the number of CAG units, and thus their melting and secondary structure characteristics. We have shown that under optimal conditions amplification efficiency of two HUMARA alleles is near-equivalent, generating PCR products in a ratio proportional to that of the genomic template. In contrast, reduction of template quantity, damage of template by ultraviolet irradiation or addition of monovalent salts (sodium chloride, sodium acetate or ammonium acetate) produces highly variable imbalances of allelic PCR products, with a strong tendency to preferentially amplify lower molecular weight alleles. Variability and biasing was diminished by substitution of 7-deaza-2'-dGTP for dGTP during amplification, an intervention which reduces stability of intramolecular and intermolecular GC base pairing. We conclude that DNA which is scanty, damaged or salt contaminated may display amplification bias of GC-rich PCR targets, potentially confounding accurate interpretation or reproducibility of assays which require co-amplification of alleles.

  4. Hierarchical modeling of genome-wide Short Tandem Repeat (STR) markers infers native American prehistory.

    Science.gov (United States)

    Lewis, Cecil M

    2010-02-01

    This study examines a genome-wide dataset of 678 Short Tandem Repeat loci characterized in 444 individuals representing 29 Native American populations as well as the Tundra Netsi and Yakut populations from Siberia. Using these data, the study tests four current hypotheses regarding the hierarchical distribution of neutral genetic variation in native South American populations: (1) the western region of South America harbors more variation than the eastern region of South America, (2) Central American and western South American populations cluster exclusively, (3) populations speaking the Chibchan-Paezan and Equatorial-Tucanoan language stock emerge as a group within an otherwise South American clade, (4) Chibchan-Paezan populations in Central America emerge together at the tips of the Chibchan-Paezan cluster. This study finds that hierarchical models with the best fit place Central American populations, and populations speaking the Chibchan-Paezan language stock, at a basal position or separated from the South American group, which is more consistent with a serial founder effect into South America than that previously described. Western (Andean) South America is found to harbor similar levels of variation as eastern (Equatorial-Tucanoan and Ge-Pano-Carib) South America, which is inconsistent with an initial west coast migration into South America. Moreover, in all relevant models, the estimates of genetic diversity within geographic regions suggest a major bottleneck or founder effect occurring within the North American subcontinent, before the peopling of Central and South America. 2009 Wiley-Liss, Inc.

  5. Assessing the genetic relationships of Curcuma alismatifolia varieties using simple sequence repeat markers.

    Science.gov (United States)

    Taheri, S; Abdullah, T L; Abdullah, N A P; Ahmad, Z; Karimi, E; Shabanimofrad, M R

    2014-09-05

    The genus Curcuma is a member of the ginger family (Zingiberaceae) that has recently become popular for use as flowering pot plants, both indoors and as patio and landscape plants. We used PCR-based molecular markers (SSRs) to elucidate genetic variation and relationships between five varieties of Curcuma (Curcuma alismatifolia) cultivated in Malaysia. Of the primers tested, 8 (of 17) SSR primers were selected for their reproducibility and high rates of polymorphism. The number of presumed alleles revealed by the SSR analysis ranged from two to six alleles, with a mean value of 3.25 alleles per locus. The values of HO and HE ranged from 0 to 0.8 (mean value of 0.2) and 0.1837 to 0.7755 (mean value of 0.5102), respectively. Eight SSR primers yielded 26 total amplified fragments and revealed high rates of polymorphism among the varieties studied. The polymorphic information content varied from 0.26 to 0.73. Dice's similarity coefficient was calculated for all pairwise comparisons and used to construct an unweighted pair group method with arithmetic average (UPGMA) dendrogram. Similarity coefficient values from 0.2105 to 0.6667 (with an average of 0.4386) were found among the five varieties examined. A cluster analysis of data using a UPGMA algorithm divided the five varieties/hybrids into 2 groups.

  6. Expressed sequence tags (ESTs and simple sequence repeat (SSR markers from octoploid strawberry (Fragaria × ananassa

    Directory of Open Access Journals (Sweden)

    Bies Dawn H

    2005-06-01

    Full Text Available Abstract Background Cultivated strawberry (Fragaria × ananassa represents one of the most valued fruit crops in the United States. Despite its economic importance, the octoploid genome presents a formidable barrier to efficient study of genome structure and molecular mechanisms that underlie agriculturally-relevant traits. Many potentially fruitful research avenues, especially large-scale gene expression surveys and development of molecular genetic markers have been limited by a lack of sequence information in public databases. As a first step to remedy this discrepancy a cDNA library has been developed from salicylate-treated, whole-plant tissues and over 1800 expressed sequence tags (EST's have been sequenced and analyzed. Results A putative unigene set of 1304 sequences – 133 contigs and 1171 singlets – has been developed, and the transcripts have been functionally annotated. Homology searches indicate that 89.5% of sequences share significant similarity to known/putative proteins or Rosaceae ESTs. The ESTs have been functionally characterized and genes relevant to specific physiological processes of economic importance have been identified. A set of tools useful for SSR development and mapping is presented. Conclusion Sequences derived from this effort may be used to speed gene discovery efforts in Fragaria and the Rosaceae in general and also open avenues of comparative mapping. This report represents a first step in expanding molecular-genetic analyses in strawberry and demonstrates how computational tools can be used to optimally mine a large body of useful information from a relatively small data set.

  7. Genetic diversity analysis with ISSR PCR on green algae Chlorella vulgaris and Chlorella pyrenoidosa

    Science.gov (United States)

    Shen, Songdong

    2008-11-01

    In the present study, genetic polymorphism and diversity in unicellular clones of Chlorella vulgaris Beijerinck and Chlorella pyrenoidosa Chick were studied with Inter Simple Sequence Repeats PCR (ISSR PCR). Samples including four clones of C. vulgaris and three clones of C. pyrenoidosa were purified by single-clone-choice method. For four C. vulgaris unicellular clones, the total number of the bands scored for 18 primers was 298; and the number of the polymorphic bands was 118, of which 39.6% were polymorphic. The size of PCR products ranged from 200 to 2 500 bp. The total number of bands scored for 18 primers, the number of polymorphic bands and the percentage of three C. pyrenoidosa unicellular clones was 194.83 and 30.8%, respectively. POPGENE analysis show that the average Nei genetic diversity (h*) and Shannon index of diversity (I*) in the four C. vulgaris unicellular clones was 0.2181 and 0.3208, respectively, which is slightly higher than those of the three C. pyrenoidosa unicellular clones (0.190 3 and 0.274 8), which agreed with the percentage of polymorphic bands in the mixed samples of the two species. The results suggest that ISSR is a useful method to Chlorella for intraspecies genetic analysis.

  8. Genetic diversity analysis with ISSR PCR on green algae Chlorella vulgaris and Chlorella pyrenoidosa

    Institute of Scientific and Technical Information of China (English)

    SHEN Songdong

    2008-01-01

    In the present study, genetic polymorphism and diversity in unicellular clones of Chlorella vulgaris Beijerinck and Chlorella pyrenoidosa Chick were studied with Inter Simple Sequence Repeats PCR (ISSR PCR). Samples including four clones of C. vulgaris and three clones of C. pyrenoidosa were purified by single-clone-choice method. For four C. vulgaris unicellular clones, the total number of the bands scored for 18 primers was 298; and the number of the polymorphic bands was 118, of which 39.6% were polymorphic. The size of PCR products ranged from 200 to 2 500 bp. The total number of bands scored for 18 primers, the number of polymorphic bands and the percentage of three C. pyrenoidosa unicellular clones was 194.83 and 30.8%, respectively. POPGENE analysis show that the average Nei genetic diversity (h *) and Shannon index of diversity (I *) in the four C. vulgaris unicellular clones was 0.2181 and 0.3208, respectively, which is slightly higher than those of the three C. pyrenoidosa unicellular clones (0.190 3 and 0.274 8), which agreed with the percentage of polymorphic bands in the mixed samples of the two species. The results suggest that ISSR is a useful method to Chlorella for intra-species genetic analysis.

  9. Identification of an avirulent Entamoeba histolytica strain with unique tRNA-linked short tandem repeat markers.

    Science.gov (United States)

    Escueta-de Cadiz, Aleyla; Kobayashi, Seiki; Takeuchi, Tsutomu; Tachibana, Hiroshi; Nozaki, Tomoyoshi

    2010-03-01

    Highly polymorphic, non-coding short tandem repeats (STR) are scattered between the tRNA genes in Entamoeba histolytica in a unique tandemly arrayed organization. STR markers that correlate with the virulence of individual E. histolytica strains have recently been reported. Here we evaluated the usefulness of tRNA-linked STR loci as genetic markers in identifying virulent and avirulent strains of E. histolytica from 37 Japanese E. histolytica samples (12 diarrheic/dysenteric, 20 amebic liver abscess (ALA), and 5 asymptomatic cases). Twenty three genotypes, assigned by combining the STR sequence types from all 6 STR loci, were identified. One to 8 new STR sequence types per locus were also discovered. Genotypes found in asymptomatic isolates were highly polymorphic (4 out of 5 genotypes were unique to this group), while in symptomatic isolates, almost half of the genotypes were shared between diarrhea/dysentery and ALA. One asymptomatic isolate (KU27) showed unique STR patterns in 4 loci. This strain, though associated with the typical pathogenic zymodeme II, failed to induce amebic liver abscess by animal challenge, which suggests that inherently avirulent E. histolytica strains exist, that are associated with unique genotypes. Furthermore, STR genotyping and in vivo challenge of 2 other asymptomatic isolates (KU14 and KU26) verified the covert virulence of these strains.

  10. An improved micropropagation of Arnebia hispidissima (Lehm.) DC. and assessment of genetic fidelity of micropropagated plants using DNA-based molecular markers.

    Science.gov (United States)

    Phulwaria, Mahendra; Rai, Manoj K; Shekhawat, N S

    2013-07-01

    An efficient and improved in vitro propagation method has been developed for Arnebia hispidissima, a medicinally and pharmaceutically important plant species of arid and semiarid regions. Nodal segments (3-4 cm) with two to three nodes obtained from field grown plants were used as explants for shoot proliferation. Murashige and Skoog's (MS) medium supplemented with cytokinins with or without indole-3-acetic acid (IAA) or naphthalene acetic acid was used for shoot multiplication. Out of different PGRs combinations, MS medium containing 0.5 mg l(-1) 6-benzylaminopurine and 0.1 mg l(-1) IAA was optimal for shoot multiplication. On this medium, explants produced the highest number of shoots (47.50 ± 0.38). About 90 % of shoots rooted ex vitro on sterile soilrite under the greenhouse condition when the base (2-4 mm) of shoots was treated with 300 mg l(-1) of indole-3-butyric acid for 5 min. The plantlets were hardened successfully in the greenhouse with 85-90 % survival rate. Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers were employed to assess the genetic stability of in vitro-regenerated plants of A. hispidissima. Out of 40 (25 RAPD and 15 ISSR) primers screened, 15 RAPD and 7 ISSR primers produced a total number of 111 (77 RAPD and 34 ISSR) reproducible amplicons. The amplified products were monomorphic across all the micropropagated plants and were similar to the mother plant. To the best of our knowledge, it is the first report on the assessment of the genetic fidelity in micropropagated plants of A. hispidissima.

  11. Evaluation of genetic homogeneity in tissue culture regenerates of Jatropha curcas L. using flow cytometer and DNA-based molecular markers.

    Science.gov (United States)

    Rathore, Mangal S; Yadav, P; Mastan, Shaik G; Prakash, Ch R; Singh, A; Agarwal, Pradeep K

    2014-01-01

    The present investigation aimed to evaluate the reliability of in vitro propagation methods for elite genotypes of Jatropha curcas L., that maintain genetic integrity of tissue culture (TC) regenerates among two regeneration systems developed through direct shoot bud regeneration using nodal/apical shoot segments (protocol-A) and in vitro-derived leaves (protocol-B) as explants. Random amplified polymorphic DNA (RAPD), intersimple sequence repeat (ISSR), simple sequence repeat (SSR) molecular markers, and flow cytometery (FCM) were employed to evaluate genetic homogeneity in TC-regenerates at different passages of subcultures. RAPD markers showed genetic homogeneity in fifth-generation TC-regenerates of both protocols. ISSR markers showed genetic stability of leaf regenerates (protocol-B) at 10th generation. FCM analysis of TC-regenerates at 10th generation in protocol-B and at 20th generation in both protocols, showed stability of ploidy level. SSR assessment of TC-regenerates at 20th generation in both protocols confirmed genetic homogeneity. The results confirmed the genetic stability of the TC-regenerates and demonstrated the reliability of the regeneration systems developed so far using explants of two different origins, for large-scale multiplication of elite genotypes of Jatropha.

  12. Molecular Genetic Diversity of Date (Phoenix dactylifera) Germplasm in Qatar based on Microsatellite Markers

    KAUST Repository

    Ahmed, Talaat

    2016-01-25

    Depending on morphological traits alone, studying the genetic diversity of date palm is a very difficult task since morphological characteristics are highly affected by the environment. DNA markers are excellent option that can help and enhance the discriminatory power of morphological characteristics. To study the genetic diversity among date palm cultivars grown in Qatar, fifteen Date palm samples were collected from Qatar University Experimental Farm. DNAs were extracted from fresh leaves by using commercial DNeasy Plant System Kit (Qiagen, Inc., Valencia, CA). Total of 18 (Inter Simple Sequence Repeat) ISSR single primers were used to amplify DNA fragments using genomic DNA of the 15 samples. First screening was done to test the ability of these primers to amplify clear bands using Date palm genomic DNA. All 18 ISSR primers successfully produced clear bands in the first screening. Then, each primer was used separately to genotype the whole set of 15 Date palm samples. Total of 4794 bands were generated using 18 ISSR primers for the 15 Date palm samples. On average, each primer generated 400 bands. The Number of amplified bands varied from cultivar to cultivar. The highest number of bands was obtained using Primers 2, 5 and 12 for the 15 (470 bands), while the lowest number of bands were obtained by Primers 1, 7 and 8 where they produced only 329 bands. Markers were scored for the presence and absence of the corresponding band among the different cultivars. Data were subjected to cluster analysis. A similarity matrix was constructed and the similarity values were used for cluster analysis.

  13. An unusual case 0020 in paternity testing: nineteen autosomal short tandem repeat typing and 12 X-chromosome markers could not clarify the case.

    Science.gov (United States)

    Tabrizi, Arash Alipour; Hejazi, Arya; Hosseini, Marzieh

    2013-12-01

    We introduce a case of disputed parentage with 2 presumptive related fathers, although using multiple genetic systems, neither of the 2 fathers may be excluded. Nineteen autosomal short tandem repeat typing and 12 X-chromosome markers could not clarify the case. We can conclude that forensic autosomic short tandem repeats included in commercial kits are not sufficient to definitively discriminate parent-offspring with related putative fathers in forensic laboratories, and supplementary investigations should be available for selected cases.

  14. Development and characterization of 1,827 expressed sequence tag-derived simple sequence repeat markers for ramie (Boehmeria nivea L. Gaud.

    Directory of Open Access Journals (Sweden)

    Touming Liu

    Full Text Available Ramie (Boehmeria nivea L. Gaud is one of the most important natural fiber crops, and improvement of fiber yield and quality is the main goal in efforts to breed superior cultivars. However, efforts aimed at enhancing the understanding of ramie genetics and developing more effective breeding strategies have been hampered by the shortage of simple sequence repeat (SSR markers. In our previous study, we had assembled de novo 43,990 expressed sequence tags (ESTs. In the present study, we searched these previously assembled ESTs for SSRs and identified 1,685 ESTs (3.83% containing 1,878 SSRs. Next, we designed 1,827 primer pairs complementary to regions flanking these SSRs, and these regions were designated as SSR markers. Among these markers, dinucleotide and trinucleotide repeat motifs were the most abundant types (36.4% and 36.3%, respectively, whereas tetranucleotide, pentanucleotide, and hexanucleotide motifs represented <10% of the markers. The motif AG/CT was the most abundant, accounting for 28.74% of the markers. One hundred EST-SSR markers (97 SSRs located in genes encoding transcription factors and 3 SSRs in genes encoding cellulose synthases were amplified using polymerase chain reaction for detecting 24 ramie varieties. Of these 100 markers, 98 markers were successfully amplified and 81 markers were polymorphic, with 2-6 alleles among the 24 varieties. Analysis of the genetic diversity of all 24 varieties revealed similarity coefficients that ranged from 0.51 to 0.80. The EST-SSRs developed in this study represent the first large-scale development of SSR markers for ramie. These SSR markers could be used for development of genetic and physical maps, quantitative trait loci mapping, genetic diversity studies, association mapping, and cultivar fingerprinting.

  15. Analysis of expressed sequence tags from Prunus mume flower and fruit and development of simple sequence repeat markers

    Directory of Open Access Journals (Sweden)

    Gao Zhihong

    2010-07-01

    Full Text Available Abstract Background Expressed Sequence Tag (EST has been a cost-effective tool in molecular biology and represents an abundant valuable resource for genome annotation, gene expression, and comparative genomics in plants. Results In this study, we constructed a cDNA library of Prunus mume flower and fruit, sequenced 10,123 clones of the library, and obtained 8,656 expressed sequence tag (EST sequences with high quality. The ESTs were assembled into 4,473 unigenes composed of 1,492 contigs and 2,981 singletons and that have been deposited in NCBI (accession IDs: GW868575 - GW873047, among which 1,294 unique ESTs were with known or putative functions. Furthermore, we found 1,233 putative simple sequence repeats (SSRs in the P. mume unigene dataset. We randomly tested 42 pairs of PCR primers flanking potential SSRs, and 14 pairs were identified as true-to-type SSR loci and could amplify polymorphic bands from 20 individual plants of P. mume. We further used the 14 EST-SSR primer pairs to test the transferability on peach and plum. The result showed that nearly 89% of the primer pairs produced target PCR bands in the two species. A high level of marker polymorphism was observed in the plum species (65% and low in the peach (46%, and the clustering analysis of the three species indicated that these SSR markers were useful in the evaluation of genetic relationships and diversity between and within the Prunus species. Conclusions We have constructed the first cDNA library of P. mume flower and fruit, and our data provide sets of molecular biology resources for P. mume and other Prunus species. These resources will be useful for further study such as genome annotation, new gene discovery, gene functional analysis, molecular breeding, evolution and comparative genomics between Prunus species.

  16. Genetic Diversity among Parents of Hybrid Rice Based on Cluster Analysis of Morphological Traits and Simple Sequence Repeat Markers

    Institute of Scientific and Technical Information of China (English)

    WANG Sheng-jun; LU Zuo-mei; WAN Jian-min

    2006-01-01

    The genetic diversity of 41 parental lines popularized in commercial hybrid rice production in China was studied by using cluster analysis of morphological traits and simple sequence repeat (SSR) markers. Forty-one entries were assigned into two clusters (I.e. Early or medium-maturing cluster; medium or late-maturing cluster) and further assigned into six sub-clusters based on morphological trait cluster analysis. The early or medium-maturing cluster was composed of 15 maintainer lines, four early-maturing restorer lines and two thermo-sensitive genic male sterile lines, and the medium or late-maturing cluster included 16 restorer lines and 4 medium or late-maturing maintainer lines. Moreover, the SSR cluster analysis classified 41 entries into two clusters (I.e. Maintainer line cluster and restorer line cluster) and seven sub-clusters. The maintainer line cluster consisted of all 19 maintainer lines, two thermo-sensitive genic male sterile lines, while the restorer line cluster was composed of all 20 restorer lines. The SSR analysis fitted better with the pedigree information. From the views on hybrid rice breeding, the results suggested that SSR analysis might be a better method to study the diversity of parental lines in indica hybrid rice.

  17. Evaluation of Population Structure, Genetic Diversity and Origin of Northeast Asia Weedy Rice Based on Simple Sequence Repeat Markers

    Directory of Open Access Journals (Sweden)

    Li Mao-bai

    2015-07-01

    Full Text Available Weedy rice exerts a severe impact on rice production by competing for sunlight, water and nutrients. This study assayed the population structure, genetic diversity and origin of Northeast Asia weedy rice by using 48 simple sequence repeat markers. The results showed that weedy rice in Northeast Asia had a high genetic diversity, with Shannon's diversity index (I of 0.748 and the heterozygosity (He of 0.434. In each regional population, I value varied widely. The widest range of I (0.228–0.489 was observed in the weedy rice of Eastern China, which was larger than that of Northeast China and Korea (0.168–0.270. The F-statistics of regional populations (Fis, Fit and Fst also showed higher values in the weedy rice of Eastern China than those of Northeast China and Korea. All weedy rice accessions were grouped into two clusters in the unweighted pair group method with arithmetic mean cluster analysis dendrogram, namely Eastern China branch and Northeastern China plus Korea branch. There was significant differentiation in genetic characteristics in weedy rice of northeastern and eastern Asia, especially in Eastern China.

  18. An integrated genetic linkage map of watermelon and genetic diversity based on single nucleotide polymorphism (SNP) and simple sequence repeat (SSR) markers

    Science.gov (United States)

    Watermelon (Citrullus lanatus var. lanatus) is an important vegetable fruit throughout the world. A high number of single nucleotide polymorphism (SNP) and simple sequence repeat (SSR) markers should provide large coverage of the watermelon genome and high phylogenetic resolution of germplasm acces...

  19. Sequence-Related Amplified Polymorphism (SRAP Markers: A Potential Resource for Studies in Plant Molecular Biology

    Directory of Open Access Journals (Sweden)

    Daniel W. H. Robarts

    2014-07-01

    Full Text Available In the past few decades, many investigations in the field of plant biology have employed selectively neutral, multilocus, dominant markers such as inter-simple sequence repeat (ISSR, random-amplified polymorphic DNA (RAPD, and amplified fragment length polymorphism (AFLP to address hypotheses at lower taxonomic levels. More recently, sequence-related amplified polymorphism (SRAP markers have been developed, which are used to amplify coding regions of DNA with primers targeting open reading frames. These markers have proven to be robust and highly variable, on par with AFLP, and are attained through a significantly less technically demanding process. SRAP markers have been used primarily for agronomic and horticultural purposes, developing quantitative trait loci in advanced hybrids and assessing genetic diversity of large germplasm collections. Here, we suggest that SRAP markers should be employed for research addressing hypotheses in plant systematics, biogeography, conservation, ecology, and beyond. We provide an overview of the SRAP literature to date, review descriptive statistics of SRAP markers in a subset of 171 publications, and present relevant case studies to demonstrate the applicability of SRAP markers to the diverse field of plant biology. Results of these selected works indicate that SRAP markers have the potential to enhance the current suite of molecular tools in a diversity of fields by providing an easy-to-use. highly variable marker with inherent biological significance.

  20. Genetic variation in Rhodomyrtus tomentosa (Kemunting) populations from Malaysia as revealed by inter-simple sequence repeat markers.

    Science.gov (United States)

    Hue, T S; Abdullah, T L; Abdullah, N A P; Sinniah, U R

    2015-12-14

    Kemunting (Rhodomyrtus tomentosa) from the Myrtaceae family, is native to Malaysia. It is widely used in traditional medicine to treat various illnesses and possesses significant antibacterial properties. In addition, it has great potential as ornamental in landscape design. Genetic variability studies are important for the rational management and conservation of genetic material. In the present study, inter-simple sequence repeat markers were used to assess the genetic diversity of 18 R. tomentosa populations collected from ten states of Peninsular Malaysia. The 11 primers selected generated 173 bands that ranged in size from 1.6 kb to 130 bp, which corresponded to an average of 15.73 bands per primer. Of these bands, 97.69% (169 in total) were polymorphic. High genetic diversity was documented at the species level (H(T) = 0.2705; I = 0.3973; PPB = 97.69%) but there was a low diversity at population level (H(S) = 0.0073; I = 0 .1085; PPB = 20.14%). The high level of genetic differentiation revealed by G(ST) (73%) and analysis of molecular variance (63%), together with the limited gene flow among population (N(m) = 0.1851), suggests that the populations examined are isolated. Results from an unweighted pair group method with arithmetic mean dendrogram and principal coordinate analysis clearly grouped the populations into two geographic groups. This clear grouping can also be demonstrated by the significant Mantel test (r = 0.581, P = 0.001). We recommend that all the R. tomentosa populations be preserved in conservation program.

  1. Modification of hippocampal markers of synaptic plasticity by memantine in animal models of acute and repeated restraint stress: implications for memory and behavior.

    Science.gov (United States)

    Amin, Shaimaa Nasr; El-Aidi, Ahmed Amro; Ali, Mohamed Mostafa; Attia, Yasser Mahmoud; Rashed, Laila Ahmed

    2015-06-01

    Stress is any condition that impairs the balance of the organism physiologically or psychologically. The response to stress involves several neurohormonal consequences. Glutamate is the primary excitatory neurotransmitter in the central nervous system, and its release is increased by stress that predisposes to excitotoxicity in the brain. Memantine is an uncompetitive N-methyl D-aspartate glutamatergic receptors antagonist and has shown beneficial effect on cognitive function especially in Alzheimer's disease. The aim of the work was to investigate memantine effect on memory and behavior in animal models of acute and repeated restraint stress with the evaluation of serum markers of stress and the expression of hippocampal markers of synaptic plasticity. Forty-two male rats were divided into seven groups (six rats/group): control, acute restraint stress, acute restraint stress with Memantine, repeated restraint stress, repeated restraint stress with Memantine and Memantine groups (two subgroups as positive control). Spatial working memory and behavior were assessed by performance in Y-maze. We evaluated serum cortisol, tumor necrotic factor, interleukin-6 and hippocampal expression of brain-derived neurotrophic factor, synaptophysin and calcium-/calmodulin-dependent protein kinase II. Our results revealed that Memantine improved spatial working memory in repeated stress, decreased serum level of stress markers and modified the hippocampal synaptic plasticity markers in both patterns of stress exposure; in ARS, Memantine upregulated the expression of synaptophysin and brain-derived neurotrophic factor and downregulated the expression of calcium-/calmodulin-dependent protein kinase II, and in repeated restraint stress, it upregulated the expression of synaptophysin and downregulated calcium-/calmodulin-dependent protein kinase II expression.

  2. Development of genome-wide informative simple sequence repeat markers for molecular diversity analysis in chickpea and development of web resource

    Directory of Open Access Journals (Sweden)

    SWARUP KUMAR PARIDA

    2015-08-01

    Full Text Available Development of informative polymorphic simple sequence repeat (SSR markers at a genome-wide scale is essential for efficient large-scale genotyping applications. We identified genome-wide 1835 SSRs showing polymorphism between desi and kabuli chickpea. A total of 1470 polymorphic SSR markers from diverse coding and non-coding regions of the chickpea genome were developed. These physically-mapped SSR markers exhibited robust amplification efficiency (73.9% and high intra- and inter-specific polymorphic potential (63.5%, thereby suggesting their immense use in various genomics-assisted breeding applications. The SSR markers particularly derived from intergenic and intronic sequences revealed high polymorphic potential. Using the mapped SSR markers, a wider functional molecular diversity (16-94%, mean: 68%, and parentage- and cultivar-specific admixed domestication pattern and phylogenetic relationships in a structured population of desi and kabuli chickpea genotypes was evident. The intra-specific polymorphism (47.6% and functional molecular diversity (65% potential of polymorphic SSR markers developed in our study is much higher than that of previous documentations. Finally, we have developed a user-friendly web resource, Chickpea Microsatellite Database (CMsDB; http://www.nipgr.res.in/CMsDB.html, which provides public access to the data and results reported in this study. The developed informative SSR markers can serve as a resource for various genotyping applications, including genetic enhancement studies in chickpea.

  3. Optimization of ISSR-PCR system for chenopodium album L%藜ISSR-PCR反应体系的优化

    Institute of Scientific and Technical Information of China (English)

    张恒庆; 朱立南; 杜洋; 贾俊玲; 谭琨

    2011-01-01

    In this study, the factors which affect the ISSR-PCR reaction were detected by TaKaRa PCR reaction system of Chenopodium album L. The purified template DNA was extracted from fresh young leaves, the effect of content of Mg2+,templates DNA and Taq DNA polymerase were tested. According to the results of amplifications in gradient it was determined that the optimization ISSR-PCR reaction system in rolumes of 25 mL contained 2.5 μL to XPCR buffer, 0.2 mrnol/L mixture of dNTPs,0.2 mmol/L primer, 1. % mmol/L Mg2+ ,40 ng of DNA and 1.5 unit Taq DNA Polymerase.%采用TaKaRa的PCR反应系统,以从藜(Chenopodium album L)幼叶中提取纯化后的总DNA为模板,进行UBC 859号引物的ISSR-PCR扩增实验,分别测试了镁离子、模板DNA和Taq DNA聚合酶含量对反应结果的影响,通过各因子的浓度梯度组合实验,确定了在25 mL体积的PCR反应中:酶配套缓冲液2.5μL,dNTPs0.2μmmol/L,引物0.2mmol/L,Mg2+1.5mmol/L,模板含量为40 ng,Taq DNA聚合酶为1.5U是藜最佳的ISSR-PCR反应体系.

  4. Genetic diversity of Greek Aegilops species using different types of nuclear genome markers.

    Science.gov (United States)

    Thomas, Konstantinos G; Bebeli, Penelope J

    2010-09-01

    Random Amplified Polymorphic DNA (RAPD) and Inter-Simple Sequence Repeat (ISSR) analyses were used to evaluate genetic variability and relationships of Greek Aegilops species. Thirty-eight accessions of seven Greek Aegilops species [Ae. triuncialis (genome UC), Ae. neglecta (UM), Ae. biuncialis (UM), Ae. caudata (C), Ae. comosa (M), Ae. geniculata (MU) and Ae. umbellulata (U)] as well as Triticum accessions were studied. Nineteen RAPD and ten ISSR primers yielded 344 and 170 polymorphic bands, respectively, that were used for the construction of dendrograms. Regardless of the similarity coefficient and marker type used, UPGMA placed 38 Aegilops accessions into one branch while the other branch consisted of wheat species. Within the Aegilops cluster, subgroups were identified that included species that shared the same genome or belonged to the same botanical section. Within the Triticum cluster, two robust subgroups were formed, one including diploid wheat and another including polyploid wheat. In conclusion, results showed that there is genetic diversity in the Greek Aegilops species studied, and clustering based on genetic similarities was in agreement with botanical classifications.

  5. Genetic characterization of Lithuanian honeybee lines based on ISSR polymorphism

    OpenAIRE

    Ceksteryte, Violeta; Paplauskiene, Vanda; Tamasauskiene, Diana; Pasakinskiene, Izolda; Mazeikiene, Ingrida

    2012-01-01

    International audience; This study presents the first results from the selection and evaluation of inter-simple sequence repeat markers for the genetic assessment of honeybee lines developed in Lithuania and introduced subspecies. Two Lithuania-bred lines of Apis mellifera carnica were compared to those introduced from Czech Republic and Slovenia and also to a subspecies introduced from the Caucasus (Apis mellifera caucasica) and local Buckfast hybrids. The genetic constitution was assayed wi...

  6. [Development of a sex-specific molecular marker for Japanese hop Humulus japonicus Siebold & Zucc].

    Science.gov (United States)

    Aleksandrov, O S; Divashuk, M G; Karlov, G I

    2011-08-01

    Japanese hop (Humulus japonicus Siebold & Zucc.) is a dioecious plant and a suitable model for studying the XX/XY1Y2 system of sex chromosomes. To develop a sex-specific marker, 12 RAPD and 36 ISSR markers were analyzed on the basis of pools of male and female plants identified after flowering. We were the first to identify ISSR marker K-16, which manifested stable amplification of an approximately 300-bp fragment in male plants and the absence of amplification in female plants in the populations examined. Marker effectiveness was confirmed in several Japanese hop populations of different origin.

  7. Genetic Diversity and Association Analysis for Salinity Tolerance,Heading Date and Plant Height of Barley Germplasm Using Simple Sequence Repeat Markers

    Institute of Scientific and Technical Information of China (English)

    Lilia Eleuch; Abderrazek Jilal; Stefania Grando; Salvatore Ceccarelli; Maria von Korff Schmising; Hisashi Tsujimoto; Amara Hajer; Abderrazek Daaloul; Michael Baum

    2008-01-01

    The objective of this study was to investigate the genetic diversity of barley accessions.Additionally,association trait analysis was conducted for grain yield under salinity,heading date and plant height.For this purpose,48 barley genotypes were analyzed with 22 microsatellite simple sequence repeat (SSR) markers.Four of the 22 markers (Bmac316,scssr03907,HVM67 and Bmag770) were able to differentiate all barley genotypes.Cluster and principal coordinate analysis allowed a clear grouping between countries from the same region.The genotypes used in this study have been evaluated for agronomic performance in different environments.Conducting association analysis for grain yield under salinity conditions using TASSEL software revealed a close association of the marker Bmag749 (2H,bin 13) in two different environments with common significant alleles (175,177),whereas the HVHOTR1 marker (2H,bin 3) was only significant in Sakhar_Egypt with alleles size being 158 and 161.Heading date also showed an association with scssr03907 through the common significant specific allele 111 and EBmacO415 markers in three different agro climatic locations,whereas HVCMA,scssr00103 and HVM67 were linked to heading date in the Egyptian environment only.The plant height association analysis revealed significant markers Bmag770 via the significant allele 152 and scssr09398.

  8. Identification and Mapping of Simple Sequence Repeat Markers from Common Bean (Phaseolus vulgaris L. Bacterial Artificial Chromosome End Sequences for Genome Characterization and Genetic–Physical Map Integration

    Directory of Open Access Journals (Sweden)

    Juana M. Córdoba

    2010-11-01

    Full Text Available Microsatellite markers or simple sequence repeat (SSR loci are useful for diversity characterization and genetic–physical mapping. Different in silico microsatellite search methods have been developed for mining bacterial artificial chromosome (BAC end sequences for SSRs. The overall goal of this study was genome characterization based on SSRs in 89,017 BAC end sequences (BESs from the G19833 common bean ( L. library. Another objective was to identify new SSR taking into account three tandem motif identification programs (Automated Microsatellite Marker Development [AMMD], Tandem Repeats Finder [TRF], and SSRLocator [SSRL]. Among the microsatellite search engines, SSRL identified the highest number of SSRs; however, when primer design was attempted, the number dropped due to poor primer design regions. Automated Microsatellite Marker Development software identified many SSRs with valuable AT/TA or AG/TC motifs, while TRF found fewer SSRs and produced no primers. A subgroup of 323 AT-rich, di-, and trinucleotide SSRs were selected from the AMMD results and used in a parental survey with DOR364 and G19833, of which 75 could be mapped in the corresponding population; these represented 4052 BAC clones. Together with 92 previously mapped BES- and 114 non-BES-derived markers, a total of 280 SSRs were included in the polymerase chain reaction (PCR-based map, integrating a total of 8232 BAC clones in 162 contigs from the physical map.

  9. Development of SCAR Marker Related to Summer Stress Tolerance in Tall Fescue (Festuca arundinacea

    Directory of Open Access Journals (Sweden)

    Xiaojun YUAN

    2014-06-01

    Full Text Available Summer stress tolerance (SST is one of the most important breeding objectives in tall fescue (Festuca arundinacea, an important perennial cool-season grass. However, breeding for better SST is generally complicated by the many environmental factors involved during the growing season. Utilizing the bulked segregant analysis (BSA, we were able to identify one marker related to SST from 100 inter-simple sequence repeat (ISSR markers and 800 random amplified polymorphic DNA (RAPD markers, and successfully developed a dominant sequence characterized amplified region (SCAR marker T_SC856 from the UBC856 sequence. Furthermore, the SCAR marker was tested in different clones of new populations, which were identified under complex summer stress (high temperature and humidity, Pythium blight, and brown patch, and it exhibited relatively high consistency (77% with the phenotype. We believe that with more markers obtained in the future, better efficiency is likely to be achieved in breeding for improved SST in tall fescue and possibly other species as well. Further studies that analyze the factors relating to the SCAR marker are needed.

  10. Repeatability of quantitative parameters of 18F-fluoride PET/CT and biochemical tumour and specific bone remodelling markers in prostate cancer bone metastases.

    Science.gov (United States)

    Wassberg, Cecilia; Lubberink, Mark; Sörensen, Jens; Johansson, Silvia

    2017-12-01

    18F-fluoride PET/CT exhibits high sensitivity to delineate and measure the extent of bone metastatic disease in patients with prostate cancer. 18F-fluoride PET/CT could potentially replace traditional bone scintigraphy in clinical routine and trials. However, more studies are needed to assess repeatability and biological uptake variation. The aim of this study was to perform test-retest analysis of quantitative PET-derived parameters and blood/serum bone turnover markers at the same time point. Ten patients with prostate cancer and verified bone metastases were prospectively included. All underwent two serial 18F-fluoride PET/CT at 1 h post-injection. Up to five dominant index lesions and whole-body 18F-fluoride skeletal tumour burden were recorded per patient. Lesion-based PET parameters were SUVmax, SUVmean and functional tumour volume applying a VOI with 50% threshold (FTV50%). The total skeletal tumour burden, total lesion 18F-fluoride (TLF), was calculated using a threshold of SUV of ≥15. Blood/serum biochemical bone turnover markers obtained at the time of each PET were PSA, ALP, S-osteocalcin, S-beta-CTx, 1CTP and BAP. A total of 47 index lesions and a range of 2-122 bone metastases per patient were evaluated. Median time between 18F-fluoride PET/CT was 7 days (range 6-8 days). Repeatability coefficients were for SUVmax 26%, SUVmean 24%, FTV50% for index lesions 23% and total skeletal tumour burden (TLF) 35%. Biochemical bone marker repeatability coefficients were for PSA 19%, ALP 23%, S-osteocalcin 18%, S-beta-CTx 22%, 1CTP 18% and BAP 23%. Quantitative 18F-fluoride uptake and simultaneous biochemical bone markers measurements are reproducible for prostate cancer metastases and show similar magnitude in test-retest variation.

  11. Four tropical, closely related fern species belonging to the genus Adiantum L. are genetically distinct as revealed by ISSR fingerprinting.

    Science.gov (United States)

    Korpelainen, Helena; de Britto, John; Doublet, Jérémy; Pravin, Sahaya

    2005-11-01

    The level and pattern of genetic variation was analyzed in four species of the fern genus Adiantum L., A. hispidulum Sw., A. incisum Forrsk., A. raddianum C.Presl, and A. zollingeri Mett. ex Kuhn, originating from South India, using the ISSR fingerprinting method. The populations of Adiantum possessed a considerable level of genetic variation, the diversity indices ranging from 0.284 to 0.464. Only 12% of the ISSR markers found were restricted to one species only, and 54% were detected in all four species. The analysis of molecular variance revealed that 71.1% of variation was present within populations. The proportion of variation detected among species was only 18.5% while the proportion of variation among populations within species equalled 10.4%. Despite the low level of intrageneric differentiation, the discriminant analysis and clustering of genetic distances indicated that the four Adiantum species are genetically distinct. The F(ST) values calculated for the species were low, varying from 0.089 to 0.179. No linkage disequilibrium was detected between the loci. Such low level of differentiation among populations and the presence of linkage equilibrium reflect that the life history of Adiantum ferns apparently involves common or relatively common sexuality, effective wind-dispersal of spores and outcrossing.

  12. Determination of Genetic Diversity Using 15 Simple Sequence Repeats Markers in Long Term Selected Japanese Quail Lines

    Science.gov (United States)

    Karabağ, Kemal; Balcıoğlu, Murat Soner; Karlı, Taki; Alkan, Sezai

    2016-01-01

    Japanese quail is still used as a model for poultry research because of their usefulness as laying, meat, and laboratory animals. Microsatellite markers are the most widely used molecular markers, due to their relative ease of scoring and high levels of polymorphism. The objective of the research was to determine genetic diversity and population genetic structures of selected Japanese quail lines (high body weight 1 [HBW1], HBW2, low body weight [LBW], and layer [L]) throughout 15th generations and an unselected control (C). A total of 69 individuals from five quail lines were genotyped by fifteen microsatellite markers. When analyzed profiles of the markers the observed (Ho) and expected (He) heterozygosity ranged from 0.04 (GUJ0027) to 0.64 (GUJ0087) and 0.21 (GUJ0027) to 0.84 (GUJ0037), respectively. Also, Ho and He were separated from 0.30 (L and LBW) to 0.33 (C and HBW2) and from 0.52 (HBW2) to 0.58 (L and LBW), respectively. The mean polymorphic information content (PIC) ranged from 0.46 (HBW2) to 0.52 (L). Approximately half of the markers were informative (PIC≥0.50). Genetic distances were calculated from 0.09 (HBW1 and HBW2) to 0.33 (C and L). Phylogenetic dendrogram showed that the quail lines were clearly defined by the microsatellite markers used here. Bayesian model-based clustering supported the results from the phylogenetic tree. These results reflect that the set of studied markers can be used effectively to capture the magnitude of genetic variability in selected Japanese quail lines. Also, to identify markers and alleles which are specific to the divergence lines, further generations of selection are required. PMID:27165027

  13. Genetic diversity and population structure among pea (Pisum sativum L.) cultivars as revealed by simple sequence repeat and novel genic markers.

    Science.gov (United States)

    Jain, Shalu; Kumar, Ajay; Mamidi, Sujan; McPhee, Kevin

    2014-10-01

    Field pea (Pisum sativum L.) is an important cool season legume crop widely grown around the world. This research provides a basis for selection of pea germplasm across geographical regions in current and future breeding and genetic mapping efforts for pea improvement. Eleven novel genic markers were developed from pea expressed sequence tag (EST) sequences having significant similarity with gene calls from Medicago truncatula spanning at least one intron. In this study, 96 cultivars widely grown or used in breeding programs in the USA and Canada were analyzed for genetic diversity using 31 microsatellite or simple sequence repeat (SSR) and 11 novel EST-derived genic markers. The polymorphic information content varied from 0.01-0.56 among SSR markers and 0.04-0.43 among genic markers. The results showed that SSR and EST-derived genic markers displayed one or more highly reproducible, multi-allelic, and easy to score loci ranging from 200 to 700 bp in size. Genetic diversity was assessed through unweighted neighbor-joining method, and 96 varieties were grouped into three main clusters based on the dissimilarity matrix. Four subpopulations were determined through STRUCTURE analysis with no significant geographic separation of the subpopulations. The findings of the present study can be used to select diverse genotypes to be used as parents of crosses aimed for breeding improved pea cultivars.

  14. Genetic analysis and association of simple sequence repeat markers with storage root yield, dry matter, starch and β-carotene content in sweetpotato.

    Science.gov (United States)

    Yada, Benard; Brown-Guedira, Gina; Alajo, Agnes; Ssemakula, Gorrettie N; Owusu-Mensah, Eric; Carey, Edward E; Mwanga, Robert O M; Yencho, G Craig

    2017-03-01

    Molecular markers are needed for enhancing the development of elite sweetpotato (Ipomoea batatas (L.) Lam) cultivars with a wide range of commercially important traits in sub-Saharan Africa. This study was conducted to estimate the heritability and determine trait correlations of storage root yield, dry matter, starch and β-carotene content in a cross between 'New Kawogo' × 'Beauregard'. The study was also conducted to identify simple sequence repeat (SSR) markers associated with these traits. A total of 287 progeny and the parents were evaluated for two seasons at three sites in Uganda and genotyped with 250 SSR markers. Broad sense heritability (H(2)) for storage root yield, dry matter, starch and β-carotene content were 0.24, 0.68, 0.70 and 0.90, respectively. Storage root β-carotene content was negatively correlated with dry matter (r = -0.59, P content, while storage root yield was positively correlated with dry matter (r = 0.57, P = 0.029) and starch (r = 0.41, P = 0.008) content. Through logistic regression, a total of 12, 4, 6 and 8 SSR markers were associated with storage root yield, dry matter, starch and β-carotene content, respectively. The SSR markers used in this study may be useful for quantitative trait loci analysis and selection for these traits in future.

  15. ISSR分子标记技术在香蕉分类上的应用%Application of Inter-Simple Sequence Repeat Markers in the Classification of Banana

    Institute of Scientific and Technical Information of China (English)

    刘绍钦; 黄上志; 梁张慧; 陈芸芸

    2007-01-01

    ISSR(inter simple sequence repeat,微卫星间隔)标记是一种基于微卫星序列发展起来的十分有效的分子标记. 根据ISSR的基本原理及其在其它作物上的应用,对收集到的包括3个基因型(AAA、AAB、ABB)在内的共15个香蕉样品,利用ISSR分子标记技术进行分类研究,将其分成四大类: 大蕉(ABB)、粉蕉(ABB)、龙牙蕉(AAB)、香牙蕉(AAA),并从分子水平上鉴定了农科1号香蕉的种性: 农科1号香蕉属于香牙蕉AAA群品系,与巴西香蕉的亲缘关系最近,遗传距离最小.

  16. Androgen receptor gene CAG repeat length as modifier of the association between Persistent Organohalogen Pollutant exposure markers and semen characteristics

    DEFF Research Database (Denmark)

    Giwercman, Aleksander; Rylander, Lars; Rignell-Hydbom, Anna;

    2007-01-01

    OBJECTIVES: Exposure to persistent organohalogen pollutants was suggested to impair male reproductive function. A gene-environment interaction has been proposed. No genes modifying the effect of persistent organohalogen pollutants on reproductive organs have yet been identified. We aimed...... and morphology) and DNA fragmentation index (DFI) were determined. CAG and GGN repeat lengths were determined by direct sequencing of leukocyte DNA. RESULTS: A statistically significant interaction was found between the CB-153 group and CAG repeat category in relation to sperm concentration and total sperm count...

  17. Construction of libraries enriched for sequence repeats and jumping clones, and hybridization selection for region-specific markers

    Energy Technology Data Exchange (ETDEWEB)

    Kandpal, R.P.; Kandpal, G.; Weissman, S.M. (Yale Univ. School of Medicine, New Haven, CT (United States))

    1994-01-04

    The authors describe a simple and rapid method for constructing small-insert genomic libraries highly enriched for dimeric, trimeric, and tetrameric nucleotide repeat motifs. The approach involves use of DNA inserts recovered by PCR amplification of a small-insert sonicated genomic phage library or by a single-primer PCR amplification of Mbo I-digested and adaptor-ligated genomic DNA. The genomic DNA inserts are heat denatured and hybridized to a biotinylated oligonucleotde. The biotinylated hybrids are retained on a Vectrex-avidin matrix and eluted specifically. The eluate is PCR amplified and cloned. More than 90% of the clones in a library enriched for (CA)[sub n] microsatellites with this approach contained clones with inserts containing CA repeats. They have also used this protocol for enrichment of (CAG)[sub n] and (AGAT)[sub n] sequence repeats and for Not I jumping clones. They have used the enriched libraries with an adaptation of the cDNA selection method to enrich for repeat motifs encoded in yeast artificial chromosomes.

  18. Expressed Sequence Tags Analysis and Design of Simple Sequence Repeats Markers from a Full-Length cDNA Library in Perilla frutescens (L.

    Directory of Open Access Journals (Sweden)

    Eun Soo Seong

    2015-01-01

    Full Text Available Perilla frutescens is valuable as a medicinal plant as well as a natural medicine and functional food. However, comparative genomics analyses of P. frutescens are limited due to a lack of gene annotations and characterization. A full-length cDNA library from P. frutescens leaves was constructed to identify functional gene clusters and probable EST-SSR markers via analysis of 1,056 expressed sequence tags. Unigene assembly was performed using basic local alignment search tool (BLAST homology searches and annotated Gene Ontology (GO. A total of 18 simple sequence repeats (SSRs were designed as primer pairs. This study is the first to report comparative genomics and EST-SSR markers from P. frutescens will help gene discovery and provide an important source for functional genomics and molecular genetic research in this interesting medicinal plant.

  19. Heterozygosities and genetic relationship of tea cultivars revealed by simple sequence repeat markers and implications for breeding and genetic mapping programs.

    Science.gov (United States)

    Tan, L Q; Zhang, C C; Qi, G N; Wang, L Y; Wei, K; Chen, S X; Zou, Y; Wu, L Y; Cheng, H

    2015-03-06

    Genetic maps are essential tools for quantitative trait locus analysis and marker-assisted selection breeding. In order to select parents that are highly heterozygous for genetic mapping, the heterozygosity (HS) of 24 tea cultivars (Camellia sinensis) was analyzed with 72 simple sequence repeat markers. In total, 359 alleles were obtained with an average of 4.99 per marker. The HS varied greatly from 37.5 to 71.0% with an average of 51.3%. On average, tea cultivars from Fujian Province showed a higher level of heterozygosity (59.8%) than those from Zhejiang (48.5%) and Yunnan (44.5%), and the 12 national tea cultivars were generally more heterozygous than the 12 provincial cultivars. Unweighted pair-group analysis using the arithmetic average grouping divided the 24 cultivars into 2 groups that are consistent with the morphological classification. All dual combinations of the 24 cultivars were studied to calculate the percentage of mappable markers when using pseudo-testcross mapping strategy, and results showed that this value also varied greatly from 51.4 to 90.3%. The genetic relationships and HS differences among different cultivars were discussed, and tea cultivars with high HS were recommended as cross parents for genetic mapping programs.

  20. ISSR Study on Genetic Relationship of Genus Paulownia%泡桐属植物亲缘关系的ISSR分析

    Institute of Scientific and Technical Information of China (English)

    莫文娟; 傅建敏; 乔杰; 雷莉莉; 李芳东; 袁德义; 邱乾栋

    2013-01-01

    利用ISSR分子标记对泡桐属21份种、变种及变型材料进行亲缘关系分析.结果表明:从100个ISSR引物中筛选出9个扩增产物电泳后带型清晰可辨的引物,共扩增出85条DNA片段,其中多态性带74条,多态位点百分率为87.05%,反映出泡桐属不同种质间遗传差异明显,其丰富的多态性与供试材料广泛的种质代表性有关;UPGMA法将研究材料分成毛泡桐组、白花泡桐组和川泡桐组3大组,地理分布较近的种质聚类在一起,表现出较密切的亲缘关系;在对兰考泡桐、山明泡桐、白花兰考泡桐、楸叶泡桐、圆冠泡桐、亮叶毛泡桐、毛泡桐、白花泡桐、川泡桐的亲缘关系分析及分类处理上,基本上与形态学、细胞学、同工酶的研究结果一致.最后对泡桐属的部分植物的分类问题进行讨论.同时发现部分泡桐的一些特异性条带,可以进一步转化为SCAR标记.%In this study, 21 species, varieties and forma in Paulownia were used to detect the genetic relationships by using inter-simple sequence repeats ( ISSR) markers. The results showed that; 1) A total of 85 DNA fragments were amplified using 9 out of 100 ISSR primers with unambiguous unique polymorphic bands, among which 74 DNA bands were polymorphic ( PPB =87. 05%), indicating significant genetic differentiation among Paulownia accessions tested; 2) The dendrogram was constructed with MEGA3. 1 statistical package using UPGMA (the unweighted pair-group method with arithmetic average) , with which Paulownia was roughly divided into three main groups of Paulownia tomentosa group, P. fortunei group and P. fargesii group. The Paulownia germplasms with close geographical distribution were clustered together, which illustrated there was a close relationship among them; 3) The findings of the relationship and classification were basically consistent with the morphology, cytology, isozyme study results among P. elongata, P. lamprophylla, P

  1. Lipid Profile and Oxidative Stress Markers in Wistar Rats following Oral and Repeated Exposure to Fijk Herbal Mixture

    Directory of Open Access Journals (Sweden)

    Oluyomi Stephen Adeyemi

    2014-01-01

    Full Text Available This study determined the effect of the oral and repeated administration of Fijk herbal mixture on rat biochemical and morphological parameters. Twenty-four Wistar rats were distributed into four groups of 6. Group A served as control and received oral administration of distilled water daily. The experimental groups B, C, and D were daily and orally exposed to Fijk herbal mixture at 15, 30, and 45 mg/kg, respectively. Treatments lasted for 21 days. The rats were sacrificed under mild diethyl ether anesthesia 24 hr after cessation of treatment. The blood and liver samples were collected and used for the biochemical and morphological analyses. Oral exposure to Fijk caused elevated levels of rat plasma ALT, AST, triglycerides, LDL, and MDA. In contrast, rat plasma HDL, GSH, and ALP levels were lowered by Fijk oral exposure. Also, the herbal remedy caused a dose-dependent elevation in the plasma atherogenic index. The histopathology examinations of rat liver sections revealed inimical cellular alterations caused by repeated exposure to Fijk. Study provides evidence that oral and repeated exposure to Fijk in rats raised the atherogenic index and potentiated oxidative stress as well as hepatic injury.

  2. Simple-sequence repeat markers of Cattleya coccinea (Orchidaceae), an endangered species of the Brazilian Atlantic Forest.

    Science.gov (United States)

    Novello, M; Rodrigues, J F; Pinheiro, F; Oliveira, G C X; Veasey, E A; Koehler, S

    2013-09-03

    Microsatellite markers were developed for the endangered Brazilian orchid species Cattleya coccinea to describe its genetic diversity and structure and to support conservation studies. Nine microsatellite loci were isolated and characterized using an enriched genomic library. All loci are polymorphic at least in the 2 populations sampled, except for loci Cac05 and Cac09 for the Petrópolis population. The mean number of alleles per locus was 8.8 between populations. The mean values of the observed and expected heterozygosities were 0.541 (ranging from 0 to 1) and 0.639 (ranging from 0 to 0.9), respectively. Cross-amplifications were performed in 7 additional Epidendroideae species, and at least 2 loci were successful in 3 additional Cattleya species, Epidendrum secundum, and Brasiliorchis gracilis. All markers described herein will be useful in further studies evaluating the genetic diversity, population dynamics, and conservation genetics of C. coccinea and related species.

  3. Separation of the genera in the subtribe Cassiinae (Leguminosae: Caesalpinioidae using molecular markers Separação dos gêneros na subtribo Cassiinae (Leguminosae: Caesalpinioidae utilizando marcadores moleculares

    Directory of Open Access Journals (Sweden)

    Laxmikanta Acharya

    2011-03-01

    Full Text Available Random amplified polymorphic DNA (RAPD, Inter simple sequence repeat (ISSR and Amplified fragment length polymorphism (AFLP markers were used to verify the segregation of the genus Cassia L. senso lato into three distinct genera namely Chamaecrista Moench., Senna P. Mill. and Cassia L. sensostricto Eighteen representatives of the three taxa were characterized using the molecular markers. 25 RAPD, six ISSR primers and six AFLP primer combinations resulted in the amplification of 612, 115 and 622 bands (loci respectively. Most of the loci are found to be polymorphic, showing high degrees of genetic diversity among the different taxa studied. The dendrogram constructed on the basis of the RAPD, ISSR and AFLP data using SHAN clustering, divided Cassia L. senso lato. into three different clusters as Chamaecrista Moench. Senna P. Mill. and Cassia L. senso stricto High bootstrap value revealed that all the clusters were stable and robust. It was observed from the present investigation that these genera have their identity at molecular level, which supports the elevation of the genus Cassia L. senso lato to the level of subtribe Cassiinae and segregation into three distinct genera instead of intrageneric categories.Técnicas de Random amplified polymorphic DNA (RAPD, Inter simple sequence repeat (ISSR e Amplified Fragment Length Polymorphism markers (AFLP foram utilizadas para verificar a segregação do gênero Cassia L. senso lato em três diferentes gêneros, Chamaecrista Moench., Senna P. Mill. e Cassia L. senso stricto Dezoito representantes dos três táxons foram caracterizados com o uso de marcadores moleculares: 25 RAPD, seis iniciadores ("primers" ISSR e seis AFLP combinações de iniciadores, resultando na amplificação de 612, 115 e 622 bandas (loci, respectivamente. A maioria dos loci apresentou-se como polimórfico, mostrando um alto grau de diversidade genética entre os táxons estudados. O dendrograma construído com base nos dados de

  4. The Extraction of the Genomic DNA in Bitter Gourd and Optimization of ISSR Amplified System%苦瓜基因组DNA的提取及ISSR扩增体系的优化

    Institute of Scientific and Technical Information of China (English)

    田丽波; 谷幸幸; 商桑; 杨衍

    2013-01-01

    In order to rapid acquiring high quality genomic DNA of Bitter gourd (Momordica charantia L.) and research the resistance gene molecular marker of powdery mildew (Sphaerotheca fuliginea), the paper compared the extracted genomic DNA' s yield and quality by different extracted methods and different physiological periods' leaves of bitter gourd to search the best method and the most suitable period in extracting genomic DNA from bitter gourd. The annealing temperatures of ISSR-PCR (inter-simple sequence repeat and polymerase chain reaction) was researched, and the concentrations of Mg2+, dNTPs, Taq DNA polymerase, genomic DNA and primers which effect ISSR-PCR were optimized by using orthogonal design method. The result showed that the OD260/OD280 values of the genomic DNA extracted from the top leaves of bitter gourd by improved CTAB method were 1.8-2.0 and the OD260/OD230 values were about 2.0, the electrophoretogram showed that the primary brands were clear and less degradation, the DNA samples were pure and had good quality, the effect was the best. The optimal system was 2.5 μL 10×PCR buffer, 2.5 mmol/LMgCl2, 250μmol/L dNTPs, 10 ng genomic DNA, 0.75 U Taq DNA polymerase, 0.7μmol/L primers, and the total volume of the reaction was 25μL, the most compatible annealing temperatures of primer UBC826 was 53℃. The optimal system could be employed to gain good results in repeated PCR experiments. The best extracting method was the improved CTAB method and the top tender leaf was the most fitting.%为了快速获取高质量的苦瓜基因组DNA,以便进行苦瓜白粉病抗性基因分子标记研究,比较了不同的DNA提取方法、不同部位的苦瓜叶片提取基因组DNA的产量和质量,探究了苦瓜基因组DNA提取的最佳方法和叶片的最适部位;研究了ISSR-PCR的退火温度,并采用正交设计法对影响ISSR-PCR的Mg2+、dNTPs、Taq酶、模板DNA以及引物浓度等5个因素进行了优化.结果表明,采用改良CTAB法

  5. Diurnal variations of plasma homocysteine, total antioxidant status, and biological markers of muscle injury during repeated sprint: effect on performance and muscle fatigue--a pilot study.

    Science.gov (United States)

    Hammouda, Omar; Chtourou, Hamdi; Chahed, Henda; Ferchichi, Salyma; Kallel, Choumous; Miled, Abdelhedi; Chamari, Karim; Souissi, Nizar

    2011-12-01

    The aim of this study was (i) to evaluate whether homocysteine (Hcy), total antioxidant status (TAS), and biological markers of muscle injury would be affected by time of day (TOD) in football players and (ii) to establish a relationship between diurnal variation of these biomarkers and the daytime rhythm of power and muscle fatigue during repeated sprint ability (RSA) exercise. In counterbalanced order, 12 football (soccer) players performed an RSA test (5 x[6 s of maximal cycling sprint + 24 s of rest]) on two different occasions: 07:00-08:30 h and 17:00-18:30 h. Fasting blood samples were collected from a forearm vein before and 3-5 min after each RSA test. Core temperature, rating of perceived exertion, and performances (i.e., Sprint 1, Sprint 2, and power decrease) during the RSA test were significantly higher at 17:00 than 07:00 h (p RSA test. However, biomarkers of antioxidant status' resting levels (i.e., total antioxidant status, uric acid, and total bilirubin) were higher in the morning. This TOD effect was suppressed after exercise for TAS and uric acid. In conclusion, the present study confirms diurnal variation of Hcy, selected biological markers of cellular damage, and antioxidant status in young football players. Also, the higher performances and muscle fatigue showed in the evening during RSA exercise might be due to higher levels of biological markers of muscle injury and lower antioxidant status at this TOD.

  6. Broad and narrow personality traits as markers of one-time and repeated suicide attempts: A population-based study

    Directory of Open Access Journals (Sweden)

    Vitaro Frank

    2008-03-01

    Full Text Available Abstract Background Studying personality traits with the potential to differentiate between individuals engaging in suicide attempts of different degrees of severity could help us to understand the processes underlying the link of personality and nonfatal suicidal behaviours and to identify at-risk groups. One approach may be to examine whether narrow, i.e., lower-order personality traits may be more useful than their underlying, broad personality trait dimensions. Methods We investigated qualitative and quantitative differences in broad and narrow personality traits between one-time and repeated suicide attempters in a longitudinal, population-based sample of young French Canadian adults using two multivariate regression models. Results One broad (Compulsivity: OR = 2.0; 95% CI 1.2–3.5 and one narrow personality trait (anxiousness: OR = 1.1; 95% CI 1.01–1.1 differentiated between individuals with histories of repeated and one-time suicide attempts. Affective instability [(OR = 1.1; 95% CI 1.04–1.1] and anxiousness [(OR = .92; 95% CI .88–.95], on the other hand, differentiated between nonattempters and one-time suicide attempters. Conclusion Emotional and cognitive dysregulation and associated behavioural manifestations may be associated with suicide attempts of different severity. While findings associated with narrow traits may be easier to interpret and link to existing sociobiological theories, larger effect sizes associated with broad traits such as Compulsivity may be better suited to objectives with a more clinical focus.

  7. Isolation and Characterization of Simple Sequence Repeats (SSR) Markers from the Moss Genus Orthotrichum Using a Small Throughput Pyrosequencing Machine

    Science.gov (United States)

    Sawicki, Jakub; Kwaśniewski, Mirosław; Szczecińska, Monika; Chwiałkowska, Karolina; Milewicz, Monika; Plášek, Vítězslav

    2012-01-01

    Here, we report the results of next-generation sequencing on the GS Junior system to identify a large number of microsatellites from the epiphytic moss Orthotrichum speciosum. Using a combination of a total (non-enrichment) genomic library and small-scale 454 pyrosequencing, we determined 5382 contigs whose length ranged from 103 to 5445 bp. In this dataset we identified 92 SSR (simple sequence repeats) motifs in 89 contigs. Forty-six of these had flanking regions suitable for primer design. We tested PCR amplification, reproducibility, and the level of polymorphism of 46 primer pairs for Orthotrichum speciosum using 40 individuals from two populations. As a result, the designed primers revealed 35 polymorphic loci with more than two alleles detected. This method is cost- and time-effective in comparison with traditional approaches involving cloning and sequencing. PMID:22837714

  8. Are There Linguistic Markers of Suicidal Writing That Can Predict the Course of Treatment? A Repeated Measures Longitudinal Analysis.

    Science.gov (United States)

    Brancu, Mira; Jobes, David; Wagner, Barry M; Greene, Jeffrey A; Fratto, Timothy A

    2016-07-02

    The purpose of this pilot study was to predict resolution of suicidal ideation and risk over the course of therapy among suicidal outpatients (N = 144) using a novel method for analyzing Self- verses Relationally oriented qualitative written responses to the Suicide Status Form (SSF). A content analysis software program was used to extract word counts and a repeated measures longitudinal design was implemented to assess improvement over time. Patients with primarily Relationally focused word counts were more likely to have a quicker suicide risk resolution than those with more Self-focused word counts (6-7 sessions versus 17-18 sessions). Implications of these data are discussed, including the potential for enhancing treatment outcomes using this method with individuals entering treatment.

  9. Development and Characterization of Microsatellite Markers in Brassica rapa ssp.chinensis and Transferability Among Related Species

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Simple sequence repeat (SSR) or microsatellite marker is a valuable tool for several purposes, such as mapping, fingerprinting,and breeding.In the present study, an inter-simple sequence repeat (ISSR)-PCR technique was applied for developing SSR markers in non-heading Chinese cabbage (Brassica rapa). A total of 190 SSRs were obtained. Among these, AG or CT (54.7%) was the most frequent repeat, followed by AC or GT (31.6%) of the microsatellites. The average number of the SSRs length array was 16 and 10 times, respectively. Based on the determined SSR sequences, 143 SSR primer pairs were designed to evaluate their transferabilities among the related species of Brassica. The number of alleles produced per marker averaged 2.91, and the polymorphism information content (PIC) value ranged from 0 to 0.863 with an average of 0.540. Monomorphism was observed in 16 primer pairs. The transferability percentage in CC genome was higher than in BB genome. More loci occurred in the BBCC genome. This result supported the hypothesis that BB genome was divergent from A and C genomes, and AA and CC genomes were relatively close. The polymorphic primers can be exploited for further evolution, fingerprinting, and variety identification.

  10. DNA cassette exchange in ES cells mediated by Flp recombinase: an efficient strategy for repeated modification of tagged loci by marker-free constructs.

    Science.gov (United States)

    Seibler, J; Schübeler, D; Fiering, S; Groudine, M; Bode, J

    1998-05-05

    The repeated modification of a genomic locus is a technically demanding but powerful strategy to analyze the function of a particular gene product or the role of cis-regulatory DNA elements in mammalian cells. The initial step is "tagging" a site with a selectable marker which is done by homologous recombination (HR) to modify a known locus or by random integration to study cis-regulatory elements at a reproducibly accessible genomic location. The tag is then used to target the construct of choice during an exchange step. Presented here is a novel technique in which the exchange is independent of HR and does not introduce vector sequences. It relies on our previous studies on the replacement of DNA cassettes by FLP-recombinase, whereby some common limitations can be overcome. To this end, the tag, a hygtk positive/negative selection marker, is integrated into the genome of embryonic stem (ES) cells. This marker is flanked by a wild-type Flp-recognition target (FRT) site on one end and by a modified heterospecific FRT site on the other. Successful Flp-mediated replacement of the hygtk cassette is enriched by ganciclovir (GANC) selection for cells that lack the encoded fusion protein. Thereby, the hygtk gene can be exchanged for virtually any sequence in a single efficient step without the need of introducing a positive selectable marker. The system can hence be used to analyze the function of either a gene product or regulatory sequences in ES cells or the transgenic mice derived thereof.

  11. A high-resolution map of genes, microsatellite markers, and new dinucleotide repeats from UBE1 to the GATA locus in the region Xp11.23

    Energy Technology Data Exchange (ETDEWEB)

    Kwan, Sau-Ping; Hagemann, T.L. [Rush Medical School, Chicago, IL (United States); Rosen, F.S. [Harvard Medical School, Boston, MA (United States)] [and others

    1995-09-01

    Several new genes and markers have recently been identified on the proximal short arm of the human X chromosome in the area of Xp11.23. We had previously generated at YAC contig in this region extending from UBE1 to the OATL1 locus. In this report two polymorphic dinucleotide repeats, DXS6949 and DXS6950, were isolated and characterized from the OATL1 locus. A panel of YAC deletion derivatives from the distal portion of the contig was used in conjunction with the rest of the YAC map to position the new microsatellites and order other markers localizing to this interval. The marker order was determined to be DXS1367-ZNF81-DXS6849-ZNF21-DXS6616-DXS6950-DXS6949. In the proximal region below OATL1, we have isolated a pair of YACs from the GATA locus, B1026 and C01160. Mapping within these YACs indicates the orientation of DXS1126 and DXS1240, while a cosmid near the OATL1 region reveals the overlap between the YAC contigs from the two loci. This cosmid contains the gene responsible for Wiskott-Aldrich syndrome (WAS) and localizes the disease gene between OATL1 and GATA. These data enable the expansion of the present physical map of the X chromosome from UBE1 to the GATA locus, covering a large portion of the Xp11.23 region. Genetic crossovers in Xp11.23 support the marker orientation and the position of WAS, contrary to previous reports. With the integration of both physical and genetic maps we have predicted the following marker order: Xpter-UBE1-SYN1/ARAF1/TIMP1/DXS1367-ZNF81-DXS-6849-ZNF21-DXSy6616-(OATL1, DXS6950-DXS6949)-WAS-(GATA,DXS1126)-DXS12410-Xcen. This orientation identifies DXS6949 and DXS1126 as the nearest flanking polymorphic markers for WAS and provides useful anchor positions for the analysis of other disease genes that have been localized to this area including three different retinal defects and X-linked nephrolithiasis. 39 refs., 3 figs., 1 tab.

  12. 紫斑牡丹遗传多样性的ISSR分析%Genetic Diversity of Paeonia rockii Revealed by ISSRs

    Institute of Scientific and Technical Information of China (English)

    杨美玲; 唐红

    2012-01-01

    利用ISSR标记对一个居群内的16个紫斑牡丹品种材料进行基因组多态性分析,从100条引物中筛选出15条用于紫斑牡丹的ISSR扩增.共扩增出134条带,其中多态性条带96条,多态性百分率为71.6%.根据ISSR扩增结果,利用NTSYSpc 2.10e软件进行Jaccard相似性系数分析,16个紫斑牡丹品种的遗传相似系数为0.45~0.93.UPGMA聚类分析结果显示,在遗传相似系数0.534处,16个紫斑牡丹品种材料可分为4类.第Ⅰ类中的6个紫斑牡丹品种材料中有5个为皇冠型品种,1个单瓣型,其他类中也有皇冠花型品种但未聚在Ⅰ类中;第Ⅱ类仅有1个‘粉盘托桂’,它是所选材料中唯一的托桂型品种,单独聚为一类;第Ⅲ类由不同花色、花型品种聚为一类,可见亲缘关系较近的紫斑牡丹品种性状变异性也很大;第Ⅳ类中A组品种材料均为白色,花型有单瓣型、荷花型和绣球型;B组仅有一个‘银线女’,它是所选材料中唯一的红色绣球型材料.研究表明,不同相似系数的遗传聚类划分与花色、花型之间并非完全具有相关性.%The genetic diversity of 16 Paeonia rockii lines in one population was analyzed by using inter-simple sequence repeat markers USSR). Fifteen primers selected from 100 primers were used for ISSR amplification. A total of 134 bands were generated,of which 96 bands were polymorphic bands (the percentage of polymorphic band,PPB=71. 6%). According to the results of ISSR amplification,which were analyzed to Jaccard similarity coefficient by NTSYSpc 2. l0e software, the genetic similarity coefficient varied from 0. 45 to 0. 93. The clustering dendrogram was constructed by UPGMA method. Sixteen P. rockii materials were divided into four major groups while the similarity coefficient was 0. 534. Class I included six of P. rockii materials,and five in six was crown form,left a single form species. But other classes also have crown flower varieties. Class Ⅱ had

  13. 水蛭ISSR-PCR反应体系的建立及优化%Establishment and optimization of ISSR-PCR reaction system for Hirudo nipponica

    Institute of Scientific and Technical Information of China (English)

    张香东; 刘庚; 常素慧; 白秀娟; 杨洪雁; 朱洪强

    2013-01-01

    Objective To establish and optimize the ISSR-PCR reaction system and amplification procedure for Hirudo nipponica and to provide the basis for its genetic diversity research. Methods Using Primer ISSR825, the orthogonal test design was adopted to optimize the ISSR-PCR amplification system on H. nipponica in five factors (Mg2+, dNTP, primer, Taq DNA polymerase, and template DNA) at four levels, and the suitable anneal temperature of the primer was selected. Results The suitable ISSR-PCR system (25 μL reaction volume) in H. nipponica included 10×PCR buffer (2.5 μL), Mg2+ (2.0 mmol/L), dNTP (0.25 mmol/L), primer (0.8 μmol/L), Taq DNA polymerase (2.5 U), and template DNA (2.0 ng/μL); The suitable anneal temperature was 49.7 ℃. Conclusion The established and optimized ISSR reaction system is stable and reliable, which could provide the basis for the analysis of genetic diversity, germplasm resources, and genetic relationship of H. nipponica.%目的 建立并优化水蛭ISSR-PCR反应体系及扩增程序,为水蛭进行遗传多样性研究提供依据.方法 使用引物ISSR825,采用正交优化方法对影响PCR反应的Mg2+、dNTP、引物、TaqDNA聚合酶、模板DNA进行了体系优化,同时对引物的最佳退火温度进行了选择.结果 在25 μL总体积中,其中包括10×PCR缓冲液2.5 μL、Mg2+ 2.0 mmol/L、dNTP0.25 mmol/L、引物0.8 μmol/L、Taq DNA聚合酶2.5 U、模板DNA 2.0 ng/μL,最佳退火温度为49.7℃.结论 所建立的最佳ISSR-PCR反应体系稳定可靠,可用于水蛭遗传多样性评价、不同种源鉴定及亲缘关系分析.

  14. Transferability of simple sequence repeat (SSR) markers developed in guava (Psidium guajava L.) to four Myrtaceae species.

    Science.gov (United States)

    Rai, Manoj K; Phulwaria, Mahendra; Shekhawat, N S

    2013-08-01

    Present study demonstrated the cross-genera transferability of 23 simple sequence repeat (SSR) primer pairs developed for guava (Psidium guajava L.) to four new targets, two species of eucalypts (Eucalyptus citriodora, Eucalyptus camaldulensis), bottlebrush (Callistemon lanceolatus) and clove (Syzygium aromaticum), belonging to the family Myrtaceae and subfamily Myrtoideae. Off the 23 SSR loci assayed, 18 (78.2%) gave cross-amplification in E. citriodora, 14 (60.8%) in E. camaldulensis and 17-17 (73.9%) in C. lanceolatus and S. aromaticum. Eight primer pairs were found to be transferable to all four species. The number of alleles detected at each locus ranged from one to nine, with an average of 4.8, 2.6, 4.5 and 4.6 alleles in E. citriodora, E. camaldulensis, C. lanceolatus and S. aromaticum, respectively. The high levels of cross-genera transferability of guava SSRs may be applicable for the analysis of intra- and inter specific genetic diversity of target species, especially in E. citriodora, C. lanceolatus and S. aromaticum, for which till date no information about EST-derived as well as genomic SSR is available.

  15. Genetic analysis and molecular characterization of Chinese sesame (Sesamum indicum L.) cultivars using insertion-deletion (InDel) and simple sequence repeat (SSR) markers.

    Science.gov (United States)

    Wu, Kun; Yang, Minmin; Liu, Hongyan; Tao, Ye; Mei, Ju; Zhao, Yingzhong

    2014-03-19

    Sesame is an important and ancient oil crop in tropical and subtropical areas. China is one of the most important sesame producing countries with many germplasm accessions and excellent cultivars. Domestication and modern plant breeding have presumably narrowed the genetic basis of cultivated sesame. Several modern sesame cultivars were bred with a limited number of landrace cultivars in their pedigree. The genetic variation was subsequently reduced by genetic drift and selection. Characterization of genetic diversity of these cultivars by molecular markers is of great value to assist parental line selection and breeding strategy design. Three hundred and forty nine simple sequence repeat (SSR) and 79 insertion-deletion (InDel) markers were developed from cDNA library and reduced-representation sequencing of a sesame cultivar Zhongzhi 14, respectively. Combined with previously published SSR markers, 88 polymorphic markers were used to assess the genetic diversity, phylogenetic relationships, population structure, and allele distribution among 130 Chinese sesame accessions including 82 cultivars, 44 landraces and 4 wild germplasm accessions. A total of 325 alleles were detected, with the average gene diversity of 0.432. Model-based structure analysis revealed the presence of five subgroups belonging to two main groups, which were consistent with the results from principal coordinate analysis (PCA), phylogenetic clustering and analysis of molecular variance (AMOVA). Several missing or unique alleles were identified from particular types, subgroups or families, even though they share one or both parental/progenitor lines. This report presented a by far most comprehensive characterization of the molecular and genetic diversity of sesame cultivars in China. InDels are more polymorphic than SSRs, but their ability for deciphering genetic diversity compared to the later. Improved sesame cultivars have narrower genetic basis than landraces, reflecting the effect of genetic

  16. Analysis of genetic diversity and population structure of peanut cultivars and breeding lines from China, India and the US using simple sequence repeat markers.

    Science.gov (United States)

    Wang, Hui; Khera, Pawan; Huang, Bingyan; Yuan, Mei; Katam, Ramesh; Zhuang, Weijian; Harris-Shultz, Karen; Moore, Kim M; Culbreath, Albert K; Zhang, Xinyou; Varshney, Rajeev K; Xie, Lianhui; Guo, Baozhu

    2016-05-01

    Cultivated peanut is grown worldwide as rich-source of oil and protein. A broad genetic base is needed for cultivar improvement. The objectives of this study were to develop highly informative simple sequence repeat (SSR) markers and to assess the genetic diversity and population structure of peanut cultivars and breeding lines from different breeding programs in China, India and the US. A total of 111 SSR markers were selected for this study, resulting in a total of 472 alleles. The mean values of gene diversity and polymorphic information content (PIC) were 0.480 and 0.429, respectively. Country-wise analysis revealed that alleles per locus in three countries were similar. The mean gene diversity in the US, China and India was 0.363, 0.489 and 0.47 with an average PIC of 0.323, 0.43 and 0.412, respectively. Genetic analysis using the STRUCTURE divided these peanut lines into two populations (P1, P2), which was consistent with the dendrogram based on genetic distance (G1, G2) and the clustering of principal component analysis. The groupings were related to peanut market types and the geographic origin with a few admixtures. The results could be used by breeding programs to assess the genetic diversity of breeding materials to broaden the genetic base and for molecular genetics studies.

  17. Analysis of genetic diversity and population structure of peanut cultivars and breeding lines from China, India and the US using simple sequence repeat markers

    Institute of Scientific and Technical Information of China (English)

    Hui Wang; Xinyou Zhang; Rajeev K. Varshney; Lianhui Xie; Baozhu Guo; Pawan Khera; Bingyan Huang; Mei Yuan; Ramesh Katam; Weijian Zhuang; Karen Harris-Shultz; Kim M. Moore; Albert K. Culbreath

    2016-01-01

    Cultivated peanut is grown worldwide as rich-source of oil and protein. A broad genetic base is needed for cultivar improvement. The objectives of this study were to develop highly informative simple sequence repeat (SSR) markers and to assess the genetic diversity and population structure of peanut cultivars and breeding lines from different breeding programs in China, India and the US. A total of 111 SSR markers were selected for this study, resulting in a total of 472 alleles. The mean values of gene diversity and polymorphic information content (PIC) were 0.480 and 0.429, respectively. Country-wise analysis revealed that alleles per locus in three countries were similar. The mean gene diversity in the US, China and India was 0.363, 0.489 and 0.47 with an average PIC of 0.323, 0.43 and 0.412, respectively. Genetic analysis using the STRUCTURE divided these peanut lines into two populations (P1, P2), which was consistent with the dendro-gram based on genetic distance (G1, G2) and the clustering of principal component analysis. The groupings were related to peanut market types and the geographic origin with a few admixtures. The results could be used by breeding programs to assess the genetic diversity of breeding materials to broaden the genetic base and for molecular genetics studies.

  18. Development of Highly Informative Genome-Wide Single Sequence Repeat Markers for Breeding Applications in Sesame and Construction of a Web Resource: SisatBase

    Science.gov (United States)

    Dossa, Komivi; Yu, Jingyin; Liao, Boshou; Cisse, Ndiaga; Zhang, Xiurong

    2017-01-01

    The sequencing of the full nuclear genome of sesame (Sesamum indicum L.) provides the platform for functional analyses of genome components and their application in breeding programs. Although the importance of microsatellites markers or simple sequence repeats (SSR) in crop genotyping, genetics, and breeding applications is well established, only a little information exist concerning SSRs at the whole genome level in sesame. In addition, SSRs represent a suitable marker type for sesame molecular breeding in developing countries where it is mainly grown. In this study, we identified 138,194 genome-wide SSRs of which 76.5% were physically mapped onto the 13 pseudo-chromosomes. Among these SSRs, up to three primers pairs were supplied for 101,930 SSRs and used to in silico amplify the reference genome together with two newly sequenced sesame accessions. A total of 79,957 SSRs (78%) were polymorphic between the three genomes thereby suggesting their promising use in different genomics-assisted breeding applications. From these polymorphic SSRs, 23 were selected and validated to have high polymorphic potential in 48 sesame accessions from different growing areas of Africa. Furthermore, we have developed an online user-friendly database, SisatBase (http://www.sesame-bioinfo.org/SisatBase/), which provides free access to SSRs data as well as an integrated platform for functional analyses. Altogether, the reference SSR and SisatBase would serve as useful resources for genetic assessment, genomic studies, and breeding advancement in sesame, especially in developing countries. PMID:28878802

  19. Development of Highly Informative Genome-Wide Single Sequence Repeat Markers for Breeding Applications in Sesame and Construction of a Web Resource: SisatBase

    Directory of Open Access Journals (Sweden)

    Komivi Dossa

    2017-08-01

    Full Text Available The sequencing of the full nuclear genome of sesame (Sesamum indicum L. provides the platform for functional analyses of genome components and their application in breeding programs. Although the importance of microsatellites markers or simple sequence repeats (SSR in crop genotyping, genetics, and breeding applications is well established, only a little information exist concerning SSRs at the whole genome level in sesame. In addition, SSRs represent a suitable marker type for sesame molecular breeding in developing countries where it is mainly grown. In this study, we identified 138,194 genome-wide SSRs of which 76.5% were physically mapped onto the 13 pseudo-chromosomes. Among these SSRs, up to three primers pairs were supplied for 101,930 SSRs and used to in silico amplify the reference genome together with two newly sequenced sesame accessions. A total of 79,957 SSRs (78% were polymorphic between the three genomes thereby suggesting their promising use in different genomics-assisted breeding applications. From these polymorphic SSRs, 23 were selected and validated to have high polymorphic potential in 48 sesame accessions from different growing areas of Africa. Furthermore, we have developed an online user-friendly database, SisatBase (http://www.sesame-bioinfo.org/SisatBase/, which provides free access to SSRs data as well as an integrated platform for functional analyses. Altogether, the reference SSR and SisatBase would serve as useful resources for genetic assessment, genomic studies, and breeding advancement in sesame, especially in developing countries.

  20. Development of novel simple sequence repeat markers in bitter gourd (Momordica charantia L.) through enriched genomic libraries and their utilization in analysis of genetic diversity and cross-species transferability.

    Science.gov (United States)

    Saxena, Swati; Singh, Archana; Archak, Sunil; Behera, Tushar K; John, Joseph K; Meshram, Sudhir U; Gaikwad, Ambika B

    2015-01-01

    Microsatellite or simple sequence repeat (SSR) markers are the preferred markers for genetic analyses of crop plants. The availability of a limited number of such markers in bitter gourd (Momordica charantia L.) necessitates the development and characterization of more SSR markers. These were developed from genomic libraries enriched for three dinucleotide, five trinucleotide, and two tetranucleotide core repeat motifs. Employing the strategy of polymerase chain reaction-based screening, the number of clones to be sequenced was reduced by 81 % and 93.7 % of the sequenced clones contained in microsatellite repeats. Unique primer-pairs were designed for 160 microsatellite loci, and amplicons of expected length were obtained for 151 loci (94.4 %). Evaluation of diversity in 54 bitter gourd accessions at 51 loci indicated that 20 % of the loci were polymorphic with the polymorphic information content values ranging from 0.13 to 0.77. Fifteen Indian varieties were clearly distinguished indicative of the usefulness of the developed markers. Markers at 40 loci (78.4 %) were transferable to six species, viz. Momordica cymbalaria, Momordica subangulata subsp. renigera, Momordica balsamina, Momordica dioca, Momordica cochinchinesis, and Momordica sahyadrica. The microsatellite markers reported will be useful in various genetic and molecular genetic studies in bitter gourd, a cucurbit of immense nutritive, medicinal, and economic importance.

  1. Evaluation of genetically modified sugarcane lines carrying Cry 1AC gene using molecular marker techniques.

    Science.gov (United States)

    Ismail, Roba M

    2013-01-01

    Five genetically modified insect resistant sugarcane lines harboring the Bt Cry 1AC gene to produce insecticidal proteins were compared with non-transgenic control by using three types of molecular marker techniques namely, RAPD, ISSR and AFLP. These techniques were applied on transgenic and non-transgenic plants to investigate the genetic variations, which may appear in sugarcane clones. This variation might demonstrate the genomic changes associated with the transformation process, which could change important molecular basis of various biological phenomena. Genetic variations were screened using 22 different RAPD primers, 10 ISSR primers and 13 AFLP primer combinations. Analysis of RAPD and ISSR banding patterns gave no exclusive evidence for genetic variations. Meanwhile, the percentage of polymorphic bands was 0.45% in each of RAPD and ISSR, while the polymorphism generated by AFLP analysis was 1.8%. The maximum percentage of polymorphic bands was 1.4%, 1.1% and 5.5% in RAPD, ISSR and AFLP, respectively. These results demonstrate that most transgenic lines showed genomic homogeneity and verified minor genomic changes. Dendrograms revealing the relationships among the transgenic and control plants were developed from the data of each of the three marker types.

  2. Improvement of Genomic DNA Extraction and Optimization of ISSR-PCR Amplification for Passion Fruit%西番莲基因组DNA的提取及ISSR-PCR的优化

    Institute of Scientific and Technical Information of China (English)

    吴田; 谢江; 蓝增全

    2011-01-01

    By comparing five DNA extraction methods, an efficient method suitable for ISSR-PCR of passion fruit (Passiflora edulis) was identified. The factors influencing ISSR-PCR for passion-fruit were optimized. The result showed that the modified SDS-Ⅱ procedure was most suitable for ISSR-PCR amplification for passion fruit. Detected on 2% agarose, 4.12 amplification bands on the average were observed in the ISSR-PCR products. The optimum amplification conditions for ISSR-PCR of passion fruit were 5 ng DNA template, 0. 2 mmol/L dNTPs, 0. 5 μmol/L primer, 0. 2 U Taq DNA polymerase and 1 × buffer (Mg2+ ) in 20μL reaction volumes.%对5种DNA提取方法进行比较,得到一种效率较高的、且适用于西番莲ISSR-PCR的DNA提取方法.同时,对影响西番莲ISSR-PCR的因子进行优化.结果表明:改良的SDS法2提取的DNA最适宜进行西番莲的ISSR-PCR扩增.ISSR-PCR产物在2%琼脂糖凝胶上检测,发现PCR扩增的平均条带数为4.12条.西番莲的ISSR-PCR的最优体系为20μL PCR反应液体系中含有1×buffer(Mg2+),0.2 mmol/L dNTPs,0.5 μmol/L引物,0.2 U Taq DNA聚合酶,5 ng DNA模板.

  3. PRELIMINARY STUDY ON FINGERPRINT ON SSR AND ISSR OF APOSTICHOPUS JAPONICUS%扩增仿刺参SSR和ISSR指纹技术的初步研究

    Institute of Scientific and Technical Information of China (English)

    闫晗; 侯林; 毕相东; 王雪

    2006-01-01

    利用SSR(Simple Sequence Repeats)技术和ISSR(Inter Simple Sequence Repeats)技术对仿刺参(Apostichopus Japonicus)进行了PCR扩增.探索了刺参基因组DNA的提取方法.为了得到更好的PCR扩增结果,对引物浓度、Mg2+浓度、dNTP浓度、模板浓度、Taq浓度和退火温度及循环数等方面进行了研究,从而摸索出一个适合刺参SSR和ISSR分析的反应体系和反应条件,并对PCR反应中的一些特定影响因子进行了分析.

  4. STUDY ON GERMPLASMIC RESOURCES OF LYCORIS LONGITUBA USING RAPD AND ISSR

    Directory of Open Access Journals (Sweden)

    Deng CHUANLIANG

    2006-08-01

    Full Text Available The perianth DNA extraction methods were discussed as Lycoris longituba for example. By means of RAPD and ISSR, germplasmic resources of Lycoris longituba were primarily studied. The results were as follow: by RAPD, a total of 77 discernible loci were obtained using 12 primers, of which 53 loci were polymorphic (PPB = 68.8%; by ISSR, 67 discernible loci were got using 9 primers, of which 62 loci were polymorphic (PPB = 92.5%. So, genetic diversity of Lycoris longituba was abundant, whose germplasmic resources could be stored for breeding. From UPGMA dendrogram of Lycoris longituba using RAPD or ISSR method, three Lycoris longituba types were supported with molecular evidence, which were originally distinguished by flower color. Therefore, in the future use of Lycoris longituba germplasmic resources, different varieties of Lycoris longituba could be cultivated.

  5. 不同颜色类群芒果的ISSR分析%ISSR analysis of different color groups in mango

    Institute of Scientific and Technical Information of China (English)

    张宇; 唐志鹏; 高兴; 杨培丽; 范琪祺; 黄国弟

    2014-01-01

    为给芒果新品种的定向选育提供理论依据,以具有不同果皮颜色的8个芒果品种的叶片为试材,采用ISSR标记技术,筛选出了特异性引物,找到了与芒果果皮颜色性状相关的特异性谱带,并运用NTsys2.10软件进行了聚类分析。结果表明,利用引物UBC-864,可寻找出红色芒果品种群的特有谱带;用引物UBC-840、UBC-857、UBC-864、UBC-868和UBC-890对供试的8份芒果品种资源进行聚类分析,以相似系数0.52为标准,可将供试材料分为2类,台农1号、金煌芒、金穗芒、四季蜜芒、桂热82号聚为一类,凯特芒、爱文芒和红芒6号聚为一类。ISSR标记可用于芒果果皮颜色性状分子标记辅助育种。%In order to provide a theoretical basis for breeding new cultivars of mango, taking eight mango cultivars with different peel colors as materials, some speciifc primers were selected out, some speciifc bands related with fruit color traits were ifnd out by using ISSR marker techniques, and the clustering analysis was conducted with NTsys2.10 software. The results show that some speciifc spectrum bands of the red mango cultivar group can be ifnd out by using primer UBC-864. The eight mango cultivars are cluster into two groups by the primers of UBC-840, UBC-857, UBC-864, UBC-868, UBC-890, taking similarity coefficient 0.52 as a standard. Tainong No.1, Jinhuang, Jinsui, Sijimi, Guire No.82 belong to one group, Keitt, Aiwen and Zill belong to anther group. ISSR markers can be used for molecular marker-assisted breeding for fruit peel color in mango.

  6. Trichinellosis survey in the wild boar from the Toledo mountains in south-western Spain (2007-2008): molecular characterization of Trichinella isolates by ISSR-PCR.

    Science.gov (United States)

    García-Sánchez, R N; Nogal-Ruiz, J J; Manzano-Lorenzo, R; Díaz, J M Arroyo; López, G Pérez; Ruano, F J Sánchez; Casas, A Ramiro; Bascón, C Crespo; Bolás-Fernández, F; Martínez-Fernández, A R

    2009-06-01

    In Spain, trichinellosis represents a public health problem, with an average of five outbreaks per year, wild boar meat being the main source of infection. A trichinellosis survey (2007-2008 hunting campaign) was carried out on wild boars in the Toledo Mountains (south-western Spain, EU) in the context of a surveillance programme on wildlife diseases. A total of 2216 wild boars from different locations of the region were examined. The examination was carried out by veterinarians in the local abattoir (Matadero Municipal de Toledo). The positive samples were sent to the Department of Parasitology (Facultad de Farmacia, UCM) for experimental isolation and specific identification by inter-simple sequence repeat-polymerase chain reaction (ISSR-PCR). Using this technique we identified 17 isolates as Trichinella spiralis with an electrophoretic profile indistinguishable from the T. spiralis reference strain (ISS48). We confirmed that ISSR-PCR is a robust technique for the molecular identification of Trichinella isolates. According to our results, the prevalence of T. spiralis in wild boars from the Toledo Mountains (>800 m above sea level) during the hunting season was approximately 0.77%. The prevalence of T. spiralis (100% of our observations) is a good example of the persistence of this species in sylvatic conditions (coming from the domestic cycle), if a good wild host is abundant. Our observations confirm the major prevalence of T. spiralis over T. britovi in this region, as well as the risk to human health represented by the consumption of uninspected wild boar meat.

  7. Análisis de la Variabilidad Genética entre treinta accesiones de tarwi (Lupinus mutabilis Sweet usando marcadores moleculares ISSR

    Directory of Open Access Journals (Sweden)

    Michelle C. Chirinos-Arias

    2015-01-01

    Full Text Available Con el fin de realizar el análisis de variabilidad genética inter-accesión de treinta accesiones de tarwi (L. mutabilis Sweet pertenecientes al Banco de Germoplasma del Instituto Nacional de Innovación Agraria (INIA. Se extrajo el ADN de 300 plantas, se construyeron bulks, se estandarizó el protocolo de amplificación de los marcadores moleculares Inter Simple Sequence Repeat (ISSR, de los cuales se eligió a los más polimórficos y nítidos para corrida en gel de acrilamida. Encontrándose 255 bandas con 8 iniciadores ISSR. El análisis de la variabilidad genética con estos iniciadores comprobó una alta variabilidad genética de las muestras en estudio. Observándose también un polimorfismo relativamente alto para una especie autógama como L. mutabilis. Finalmente los fenogramas mostraron una relación con la ubicación geográfica, posiblemente debido al flujo génico in situ debido al intercambio o venta de semillas en ferias o mercados aledaños a la zona de colecta.

  8. Genetic diversity analysis based on molecular marker and quantitative traits of the response of different tomato (Lycopersicon esculentum Mill. cultivars to drought stress

    Directory of Open Access Journals (Sweden)

    Metwali Ehab M.R.

    2016-01-01

    Full Text Available The drought tolerance of tomato (Lycopersicon esculentum Mill. is a trait needing urgent improvement due to recent climate changes and limited water availability. We therefore conducted a greenhouse screening experiment to identify tomato cultivars with improved drought tolerance. Several sensitivity and tolerance indices were computed based on morphological markers. With the aim of establishing a correlation to these markers, a total of 16 inter-simple sequence repeat (ISSR primers were used, the genetic diversity among cultivars was elucidated and clustering the cultivars into groups based on their molecular profiles was performed. The obtained results indicated that selection indices, such as geometric mean productivity (GMP, mean productivity (MP, tolerance index (TOL,and stress tolerance index (STI, represented suitable indices for screening the drought tolerance of tomato cultivars. An interesting correlation of the ISSR analyses to these morphological findings was established according to 83 detectable fragments derived from 10 primers. The highest value of the effective multiplex ratio (EMR and marker index (MI was detected for primer INC7 followed by INC1. Based on Jaccard's similarity coefficients, the genetic distance of the genotypes varied from 0.702 to 0.942 with a mean value of 0.882. The results showed a clear-cut separation of the 15 tomato cultivars due to their genetic variability, making them a valuable genetic source for their incorporation into potential breeding programs. Molecular data were in good agreement with the results as regards selection indices, and both of them will be useful tools for improvement of the tomato germplasm.

  9. Repeated measurements of cerebral blood flow in the left superior temporal gyrus reveal tonic hyperactivity in patients with auditory verbal hallucinations: A possible trait marker

    Directory of Open Access Journals (Sweden)

    Philipp eHoman

    2013-06-01

    Full Text Available Background: The left superior temporal gyrus (STG has been suggested to play a key role in auditory verbal hallucinations in patients with schizophrenia. Methods: Eleven medicated subjects with schizophrenia and medication-resistant auditory verbal hallucinations and 19 healthy controls underwent perfusion magnetic resonance imaging with arterial spin labeling. Three additional repeated measurements were conducted in the patients. Patients underwent a treatment with transcranial magnetic stimulation (TMS between the first 2 measurements. The main outcome measure was the pooled cerebral blood flow (CBF, which consisted of the regional CBF measurement in the left superior temporal gyrus (STG and the global CBF measurement in the whole brain.Results: Regional CBF in the left STG in patients was significantly higher compared to controls (p < 0.0001 and to the global CBF in patients (p < 0.004 at baseline. Regional CBF in the left STG remained significantly increased compared to the global CBF in patients across time (p < 0.0007, and it remained increased in patients after TMS compared to the baseline CBF in controls (p < 0.0001. After TMS, PANSS (p = 0.003 and PSYRATS (p = 0.01 scores decreased significantly in patients.Conclusions: This study demonstrated tonically increased regional CBF in the left STG in patients with schizophrenia and auditory hallucinations despite a decrease in symptoms after TMS. These findings were consistent with what has previously been termed a trait marker of auditory verbal hallucinations in schizophrenia.

  10. The Metabolic Syndrome, Inflammation, and Colorectal Cancer Risk: An Evaluation of Large Panels of Plasma Protein Markers Using Repeated, Prediagnostic Samples

    Directory of Open Access Journals (Sweden)

    Sophia Harlid

    2017-01-01

    Full Text Available Metabolic syndrome (MetS, a set of metabolic risk factors including obesity, dysglycemia, and dyslipidemia, is associated with increased colorectal cancer (CRC risk. A putative biological mechanism is chronic, low-grade inflammation, both a feature of MetS and a CRC risk factor. However, excess body fat also induces a proinflammatory state and increases CRC risk. In order to explore the relationship between MetS, body size, inflammation, and CRC, we studied large panels of inflammatory and cancer biomarkers. We included 138 participants from the Västerbotten Intervention Programme with repeated sampling occasions, 10 years apart. Plasma samples were analyzed for 178 protein markers by proximity extension assay. To identify associations between plasma protein levels and MetS components, linear mixed models were fitted for each protein. Twelve proteins were associated with at least one MetS component, six of which were associated with MetS score. MetS alone was not related to any protein. Instead, BMI displayed by far the strongest associations with the biomarkers. One of the 12 MetS score-related proteins (FGF-21, also associated with BMI, was associated with an increased CRC risk (OR 1.71, 95% CI 1.19–2.47. We conclude that overweight and obesity, acting through both inflammation and other mechanisms, likely explain the MetS-CRC connection.

  11. Analysis of genetic diversity and population structure of oil palm (Elaeis guineensis) from China and Malaysia based on species-specific simple sequence repeat markers.

    Science.gov (United States)

    Zhou, L X; Xiao, Y; Xia, W; Yang, Y D

    2015-12-08

    Genetic diversity and patterns of population structure of the 94 oil palm lines were investigated using species-specific simple sequence repeat (SSR) markers. We designed primers for 63 SSR loci based on their flanking sequences and conducted amplification in 94 oil palm DNA samples. The amplification result showed that a relatively high level of genetic diversity was observed between oil palm individuals according a set of 21 polymorphic microsatellite loci. The observed heterozygosity (Ho) was 0.3683 and 0.4035, with an average of 0.3859. The Ho value was a reliable determinant of the discriminatory power of the SSR primer combinations. The principal component analysis and unweighted pair-group method with arithmetic averaging cluster analysis showed the 94 oil palm lines were grouped into one cluster. These results demonstrated that the oil palm in Hainan Province of China and the germplasm introduced from Malaysia may be from the same source. The SSR protocol was effective and reliable for assessing the genetic diversity of oil palm. Knowledge of the genetic diversity and population structure will be crucial for establishing appropriate management stocks for this species.

  12. Assessment of genetic relationship in Persea spp by traditional molecular markers.

    Science.gov (United States)

    Reyes-Alemán, J C; Valadez-Moctezuma, E; Barrientos-Priego, A F

    2016-04-04

    Currently, the reclassification of the genus Persea is under discussion with molecular techniques for DNA analysis representing an alternative for inter- and intra-specific differentiation. In the present study, the traditional random-amplified polymorphic DNA (RAPD) and the inter simple sequence repeat (ISSR) markers were used to determine the genomic relationship of different species and hybrids representative of the subgenera Eriodaphne and Persea in a population conserved in a germplasm bank. The data were analyzed statistically using multivariate methods. In the RAPD analysis, a total of 190 polymorphic bands were produced, with an average of 23.7 bands per primer, the percentage contribution of each primer was from 7.66 to 19.63; the polymorphic information content (PIC) ranged from 0.23 to 0.45, with an average of 0.35. In the ISSR analysis, a total of 111 polymorphic bands were considered, with an average of 18.5 bands per primer, the percentage contribution of each was from 11.83 to 19.57; the PIC ranged from 0.35 to 0.48, with an average of 0.42. The phenograms obtained in each technique showed the relationship among the accessions through the clusters formed. In general, both the techniques grouped representatives of the Persea americana races (P. americana var. drymifolia, P. americana var. guatemalensis, and P. americana var. americana). However, it was not possible to separate the species of Persea used as reference into independent clades. In addition, they tended to separate the representatives of subgenera Eriodaphne and Persea.

  13. Genomic markers reveal introgressive hybridization in the Indo-West Pacific mangroves: a case study.

    Directory of Open Access Journals (Sweden)

    Mei Sun

    Full Text Available Biodiversity of mangrove ecosystems is difficult to assess, at least partly due to lack of genetic verification of morphology-based documentation of species. Natural hybridization, on the one hand, plays an important role in evolution as a source of novel gene combinations and a mechanism of speciation. However, on the other hand, recurrent introgression allows gene flow between species and could reverse the process of genetic differentiation among populations required for speciation. To understand the dynamic evolutionary consequences of hybridization, this study examines genomic structure of hybrids and parental species at the population level. In the Indo-West Pacific, Bruguiera is one of the dominant mangrove genera and species ranges overlap extensively with one another. Morphological intermediates between sympatric Bruguiera gymnorrhiza and Bruguiera sexangula have been reported as a variety of B. sexangula or a new hybrid species, B. × rhynchopetala. However, the direction of hybridization and extent of introgression are unclear. A large number of species-specific inter-simple sequence repeat (ISSR markers were found in B. gymnorrhiza and B. sexangula, and the additive ISSR profiling of B. × rhynchopetala ascertained its hybrid status and identified its parental origin. The varying degree of scatterness among hybrid individuals in Principal Coordinate Analysis and results from NewHybrids analysis indicate that B. × rhynchopetala comprises different generations of introgressants in addition to F(1s. High genetic relatedness between B. × rhynchopetala and B. gymnorrhiza based on nuclear and chloroplast sequences suggests preferential hybrid backcrosses to B. gymnorrhiza. We conclude that B. × rhynchopetala has not evolved into an incipient hybrid species, and its persistence can be explained by recurrent hybridization and introgression. Genomic data provide insights into the hybridization dynamics of mangrove plants. Such information

  14. Genome-Wide Analysis of Simple Sequence Repeats and Efficient Development of Polymorphic SSR Markers Based on Whole Genome Re-Sequencing of Multiple Isolates of the Wheat Stripe Rust Fungus.

    Directory of Open Access Journals (Sweden)

    Huaiyong Luo

    Full Text Available The biotrophic parasitic fungus Puccinia striiformis f. sp. tritici (Pst causes stripe rust, a devastating disease of wheat, endangering global food security. Because the Pst population is highly dynamic, it is difficult to develop wheat cultivars with durable and highly effective resistance. Simple sequence repeats (SSRs are widely used as molecular markers in genetic studies to determine population structure in many organisms. However, only a small number of SSR markers have been developed for Pst. In this study, a total of 4,792 SSR loci were identified using the whole genome sequences of six isolates from different regions of the world, with a marker density of one SSR per 22.95 kb. The majority of the SSRs were di- and tri-nucleotide repeats. A database containing 1,113 SSR markers were established. Through in silico comparison, the previously reported SSR markers were found mainly in exons, whereas the SSR markers in the database were mostly in intergenic regions. Furthermore, 105 polymorphic SSR markers were confirmed in silico by their identical positions and nucleotide variations with INDELs identified among the six isolates. When 104 in silico polymorphic SSR markers were used to genotype 21 Pst isolates, 84 produced the target bands, and 82 of them were polymorphic and revealed the genetic relationships among the isolates. The results show that whole genome re-sequencing of multiple isolates provides an ideal resource for developing SSR markers, and the newly developed SSR markers are useful for genetic and population studies of the wheat stripe rust fungus.

  15. Assessment of genetic diversity among different indigenous Xanthomonas isolates via RAPD and ISSR

    Directory of Open Access Journals (Sweden)

    Fatima Sabin

    2012-01-01

    Full Text Available The genetic diversity among seven Xanthomonas isolates representing four species was assessed using RAPD and ISSR PCR-based techniques. Both techniques revealed high degrees of polymorphisms among the studied isolates. A cluster dendrogram based on the combined data of RAPD and ISSR showed that genetic diversity exists in local isolates of Xanthomonas. In terms of percentage similarity values, the genomic variation was found to be in the range of 29.29% - 100% among the isolates. X. campestris (Mangifera indica remained unclustered in cluster dendrogram and revealed a unique genomic profile compared to other isolates used in this study.

  16. (SSR) markers

    African Journals Online (AJOL)

    SAM

    2014-07-30

    Jul 30, 2014 ... Simple sequence repeats (SSRs) are the most widely used marker system for plant variety characterization and ... gene tagging in marker assisted breeding and gene cloning in .... PLS-2 and PAU Selection Long) to 1.00 (between PC. 2062 and .... Comparative analyses of genetic diversities within tomato.

  17. Marker-Assisted Backcrossing to Develop an Elite Cytoplasmic Male Sterility Line in Rice

    Directory of Open Access Journals (Sweden)

    Asadollah Ahmadikhah

    2015-07-01

    Full Text Available Cytoplasmic male sterility (CMS is a cornerstone of hybrid production in many crops. In three-line hybrid systems, use of CMS, maintainer, and fertility restorer lines is necessary for production of hybrid seeds. Limited resources of CMS and low variation of CMS lines cause genetic vulnerability to pathogens. Therefore, diversifying the CMS sources is indispensible for a sustainable production system of hybrid seed. In this study, we attempted for the first time to transfer CMS into maintainer line Yosen B in restricted generations using the marker-assisted backcrossing (MABC method. The resultant F hybrid of IR68897 A/Yosen B cross was backcrossed to Yosen B, and CMS plants in each backcross generation (from BCF to BCF were selected based on phenotyping test and MABC. Molecular assessment of backcross progenies was conducted using a mitochondrial CMS-specific marker and 34 polymorphic nuclear simple-sequence repeat (SSR markers in early generations (from BCF to BCF and was continued using 9 additional SSRs and 82 inter-simple sequence repeat (ISSR markers in BCF. A MABC strategy could successfully recover the recurrent parent genome (RPG in BCF generation, and decreased heterozygosity of final CMS plants. Restorability test with known wild-abortive restorer lines (viz. IR36 and IR24 showed that combination of Yosen A × IR24 could produce highly fertile F hybrid. Evaluation of some important agronomic traits of the final CMS line (BCF at field condition showed that it was comparable to the original maintainer fertile counterpart. Phenotypic and marker-assisted selections could considerably decrease the time needed for full recovery of RPG so that final CMS line could show a high similarity to original fertile counterpart.

  18. Establishment of a SRAP analysis protocol in Carya cathayensis and a comparison among SRAP,RAPD,ISSR analysis protocols%山核桃SRAP体系的建立及与RAPD和ISSR标记的比较

    Institute of Scientific and Technical Information of China (English)

    李元春; 沈林; 曾燕如

    2011-01-01

    In order to have a good molecular marker to reflect inherent genetic characteristics of Lin'an hickory (Carya cathayensis), the genomic DNA extracted from hickory leaves was used to optimize parameters (constituents) included in a sequence-related amplified polymorphism-polymerase chain reaction (SRAP-PCR) protocol run under the following conditions: pre-denaturing at 94 ℃ for 5 min; 5 cycles, each of which denatured at 94 ℃ for 30 s and annealed at 35 ℃ for 30 s with an extension at 72 ℃ for 2 min; 30 cycles,each of which denatured at 94 ℃ for 30 s and annealed at 50 ℃ for 30 s with an extension at 72 ℃ for 2 min;and a final extension at 72 ℃ for 8 min.Then, 15 pairs of primers out of 100 pairs were screened for SRAP analysis and a comparison was made among SRAP, random amplified polymorphic DNA (RAPD), and intersimple sequence repeat(ISSR).The optimized(SRAP-PCR)protocol was as follows: a total volume of 25.00 μL containing 1 × buffer, 0.20 mmol· L-1 dNTPs ( deoxynucleotide triphosphates ), 0.20 μ mol· L-1 p rimers,2.00 mmol· L-1 Mg2+, 33.34 nkat Taq DNA polymerase, and 0.80 mg·L-1 genomic DNA(all at a final concentration).On the average, SRAP, compared to RAPD and ISSR, had the most loci and polymorphic loci amplified by each pair of primers, but SRAP percentages for both polymorphic primer pairs and polymorphic loci were between RAPD and ISSR.The optimized SRAP-PCR reduced the reaction time by half compared with the former protocols.It has been shown that both SRAP and RAPD should be considered when studying hickory.%以山核桃Carya cathayensis 基因组DNA为模板,对聚合酶链式反应(PCR)体系各组分进行了梯度实验,优化出条带清晰、重复性好的相关序列扩增多态性聚合酶链式反应(SRAP-PCR)扩增体系,并筛选了引物.该体系(25.00μL)为:1×缓冲液0.20 mmol·L-1,脱氧核糖核苷酸(dNTPs),0.20μmol·L-1引物,2.00mmol·L-1镁离子(Mg2+),33.34 nkat Taq DNA聚合酶,0.80 mg·L-1基因组DNA

  19. Analysis of Genetic Variation between Diploids and Their Homologous Tetraploids of lsatis indigotica Fort.by ISSR%菘蓝二倍体及其同源四倍体遗传差异的 ISSR 分析

    Institute of Scientific and Technical Information of China (English)

    段英姿

    2012-01-01

    以1个菘蓝二倍体及其10个同源四倍体株系为材料,利用19个随机扩增ISSR引物分析其遗传差异性,为菘蓝多倍体诱变的基因表达调控以及遗传改良提供依据.结果显示:菘蓝二倍体与其同源四倍体及四倍体之间的ISSR多态性有明显差异,除主要遗传位点相同外,有些四倍体株系扩增条带数多于二倍体,有些四倍体株系扩增条带数少于二倍体,四倍体株系间亦有多态位点;11个株系共扩增111条多态性条带,多态性达55.22%.聚类分析显示,不同四倍体株系与二倍体的遗传差异大小亦不同.研究表明,菘蓝二倍体与其同源四倍体具有中等偏高的遗传差异性.%ISSR molecular markers with ninteen primers were used to analyze the genetic variation between 1 diploids and their 10 homological tetraploids of Isatis indigotica Fort. . The results showed that genetic variation was distinct difference between diploids and their homological tetraploids of Isatis varied with different genotypes. In addition to the main genetic loci were same,some tetraploids amplified extra bands and some tetraploids with missing bands. There were polymorphic loci between the homological tetraploids. 11 strains were detected 111 alleles,its homological tetraploid was 55. 22%. The clustering results show that genetic variation is different between diploids and tetraploids. The genetic variation with medium is on the high side between diploids and tetraploids by ISSR.

  20. Determinación de la estructura poblacional de litopenaeus vannamei mediante issrs a lo largo de la costa ecuatoriana

    OpenAIRE

    Fresneda Rodríguez, Adriana

    2003-01-01

    Determinación de la estructura poblacional de Litopenaeus vannamei mediante ISSRs a lo largo de la costa ecuatoriana Muestras de Litopenaeus vannamei fueron colectadas en siete localidades a lo largo de la costa ecuatoriana desde Esmeraldas hasta Machala.

  1. 长春花ISSR-PCR 反应体系的正交优化%Research on the Orthogonal Optimization of ISSR-PCR Reaction System of Madagascar Periwinkle

    Institute of Scientific and Technical Information of China (English)

    刘峰; 顾志敏; 陈析丰; 金杨; 马伯军

    2010-01-01

    [目的]筛选最适的长春花ISSR-PCR 反应体系.[方法]采用正交试验方法,对影响长春花ISSR-PCR反应体系的引物浓度、dNTP浓度、Mg~(2+)浓度、Taq DNA聚合酶浓度和模板DNA浓度5个因素在4个水平上进行正交试验,建立适合长春花ISSR-PCR的反应体系.[结果]长春花ISSR-PCR 反应体系的最佳条件:引物浓度0.50 μmol/L,dNTP浓度0.10 mmol/L,Mg~(2+)浓度 3.00 mmol/L,Taq 酶浓度0.75 U/25 μl,DNA模板浓度350 ng/25 μl.采用该反应体系筛选出12对适合于长春花ISSR-PCR反应的引物.[结论]利用优化系统进行长春花的ISSR-PCR反应,可获得稳定性高、重复性好、背景清晰的电泳结果.

  2. Population Genetics of the Endemic Hawaiian Species Chrysodracon hawaiiensis and Chrysodracon auwahiensis (Asparagaceae: Insights from RAPD and ISSR Variation

    Directory of Open Access Journals (Sweden)

    Pei-Luen Lu

    2016-08-01

    Full Text Available The genus Chrysodracon has six endemic species in the Hawaii Islands. Chrysodracon hawaiiensis is endemic to Hawaii Island and was described as a distinct species in 1980. It was listed as an endangered species on the International Union for the Conservation of Nature and Natural Resources (IUCN Red List in 1997. This woody plant species was, at one time, common in exposed dry forests, but it became very rare due to grazing pressure and human development. The tree species Chrysodracon auwahiensis (C. auwahiensis, endemic to Maui and Molokai, still has large adult populations in dry lands of the islands, but unfortunately no regeneration from seed has been reported in those areas for many years. The two endemic species were examined using the molecular technique of random amplified polymorphic DNA (RAPD and inter simple sequence repeats (ISSR to determine the genetic structure of the populations and the amount of variation. Both species possess similar genetic structure. Larger and smaller populations of both species contain similar levels of genetic diversity as determined by the number of polymorphic loci, estimated heterozygosity, and Shannon’s index of genetic diversity. Although population diversity of Chrysodracon hawaiiensis (C. hawaiiensis is thought to have remained near pre-disturbance levels, population size continues to decline as recruitment is either absent or does not keep pace with senescence of mature plants. Conservation recommendations for both species are suggested.

  3. ISSR Analysis of the Genetic Diversity of the Endangered Species Sinopodophyllum hexandrum (Royle) Ying from Western Sichuan Province, China

    Institute of Scientific and Technical Information of China (English)

    Meng Xiao; Qun Li; Li Wang; Liang Guo; Jing Li; Lin Tang; Fang Chen

    2006-01-01

    Sinopodophyllum hexandrum (Royle) Ying Is an important medicinal and endangered species. Inter-simple sequence repeats (ISSR) analysis was conducted on seven natural populations from western Sichuan Province to investigate the genetic diversity of S. hexandrum. Leaf samples of 140 individuals were collected.Of the 139 discernible fragments generated by 12 selected primers (among 100 primers), 54 appeared to be polymorphic. The percentage of polymorphic bands (PPB) was 38.85% at the species level, and PPB within a population ranged from 7.91% to 23.74%. Low levels of genetic variation (He=0.092, Ho=0.142) and high levels of genetic differentiation among the populations (Gst= 62.25%) was detected on the basis of results from POPGENE and analysis of molecular variance (AMOVA), respectively. Furthermore, the limited gene flow (Nm=0.361) may result from biological characteristics, such as self-pollination and short distance seed dispersal. Based on the genetic and ecological information available for S. hexandrum, we propose some appropriate strategies for the conservation of the endangered medicinal species in this region,namely rescuing and conserving the core populations for in situ conservation and sampling and preserving more populations with fewer individuals from each population for ex situ conservation.

  4. Optimization of short tandem repeats (STR) typing method and allele frequency of 8 STR markers in referring to forensic medicine of Semnan Province.

    Science.gov (United States)

    Eskandarion, M; Najafi, M; Akbari Eidgahi, M; Alipour Tabrizi, A; Golmohamadi, T

    2015-01-01

    Background and Objective: Short Tandem Repeats (STR) show considerable differences among individuals in the population from which they used for identification. There are various methods for analysis of these STR loci, and capillary electrophoresis method already used as an international standard. Due to the high costs of this process, this study aimed to set up a Multiplex PCR method in some standard STR loci so that we can use its PCR product in STR analysis with different methods of HPLC, GC-Mass, and Capillary Electrophoresis. Materials and Methods: 8 typical STR loci in the identification selected according to their size in the two groups of four (CSF1PO, VWA, D18S51, PentaD and TPOX, Amelogenin, FGA, SE33) from NIST (National Institute of Standards and Technology). The above SSR primers prepared from Genbank and Monoplex PCR was designed based on their size. Then, with the changes in temperature conditions, magnesium ion, primers concentration, and setting-up, Hot Start Multiplex PCR of four markers was carried out. PCR product investigated on the agarose gel electrophoresis (3%) and the results of genotyping analyzed by Genetic Analyzer. Results: The Results showed that all STR loci under study are detectable as Monoplex PCR at a temperature of 62°-66° and 1.5 mM magnesium ion. Moreover, Multiplex PCR results showed that when the concentration of primer and temperature measured by the fixed concentration of magnesium, CSF1PO, and D18S51 loci bands are weaker than desired. Using a standard buffer and set Magnesium conditions against changes in the primer concentration and temperature, when Taq polymerase enzyme is added to test tubes at a temperature of 94°, Multiplex PCR bands are visible desirably. Capillary electrophoresis genotyping results obtained in all eight loci and the Locus FGA had the most allelic diversity and the loci TPOX and CSF1PO had the lowest allelic diversity. TPOX and CSF1PO loci had the lowest allelic frequencies, and FGA locus had

  5. Molecular markers associated with the agronomic traits in the medicinal plant lemon balm

    Directory of Open Access Journals (Sweden)

    Sanam Safaei-Chaeikar

    2017-06-01

    Full Text Available Finding association between molecular markers and agronomic traits provide an excellent tool for indirect selection of a trait of interest in the population. In this study, stepwise regression analysis was used to estimate associations between ISSR and RAPD markers with some agronomic traits in lemon balm ecotypes. The analysis of results revealed significant associations between the traits and some of the studied loci. For all the traits, more than one informative marker was detected. Totally,90informative markers, including 48 ISSR loci and 42 RAPD loci, were identified. The SA-R-10, UBC826-1, UBC812-9, UBC813-10, UBC825-4, OPA-01-15, OPC-04-7 and CS-56-8 markers or fragment showed a significant correlation with Essential oil percentage and controlled 99.8% of the phenotypic variation. These markers are relatively more reliable. Among the RAPD primers, special attention should be drawn to primer SA-R, which had the highest associated fragments with the traits including days for 50% flowering, number of branches per plant, fresh weight and dry weight. Some of ISSR and RAPD markers were associated with more than one trait in multiple regression analysis that may be due to pleiotropic effect of the linked quantitative trait locus on different traits or its linkage to different genes. These primers have been found useful for improved lemon balm.

  6. Genetic diversity in palmyrah genotypes using morphological and molecular markers

    Directory of Open Access Journals (Sweden)

    V.Ponnuswami

    2010-07-01

    Full Text Available Palms are woody monocotyledons in the family Arecaceae which is placed in the order Arecales. Slow and tall growing,hardy and non branching, dioecious and perennial in nature, palmyrah palm has no distinguishing features to identify sex,stature and high neera yielding types until flowering age of about 12 to 15 years. Under these circumstances molecularmarkers can be effectively utilized to diagnose and select a genotype. Twenty palmyrah accessions were analysed usingRAPD and ISSR markers. In RAPD analysis, a total of 57 bands were obtained, among them 43 were polymorphic and restof them were monomorphic. Amplification size ranged between 250 and 3200 bp. UPGMA based cluster diagram showedthat all 20 different genotypes were grouped into four different clusters based on the stature, sex and high neera yieldingtypes. The distance matrix between genotypes showed an average distance range from 0.54 to 0.91 with a mean of 0.70. Atotal of 130 ISSR markers were scored, of which 65 were polymorphic, equivalent to 47.94% polymorphism. These markerswere used to estimate the genetic similarity among accessions using Jaccard’s similarity coefficient, with similarity valuesranging from 71.6 to 95.7%. The average number of markers produced per primer was 6.11. For each of the 21 ISSRprimers PIC value ranged between 0 and 0.46. Cluster analysis based on ISSR data grouped the 20 palmyrah accessions intotwo major clusters. PCA based on ISSR data clearly distinguished genotypes similar to the results of cluster analysis.

  7. DNA extraction of birch leaves by improved CTAB method and optimization of its ISSR system

    Institute of Scientific and Technical Information of China (English)

    PAN hua; YANG Chuan-ping; WEI Zhi-gang; JIANG Jing

    2006-01-01

    The basic method of DNA extraction (CTAB) was improved as the multi-times STE-CTAB extraction method and used to extract the DNA of birch leaved in this experiment. Results showed that the improved method is suitable not only for genomic DNA extraction of birch but also for that of other plants. The purity of genomic DNA extracted by the multi-times STE-CTAB extraction method is higher than that by one time STE-CTAB method, and it does not need the process of RNase. The factors of influencing ISSR system were explored based on the genomic DNA of birch extracted by the two methods. The optimal conditions for ISSR system were determined as follows: cycles of denaturation for 30 s at 94℃, annealing for 30 s at 51 ℃, extension for 30 s at 72℃, and a final 7 min extension at 72 ℃.

  8. Genetic variation in two sea cucumber (Apostichopus japonicus) stocks revealed by ISSR markers

    Institute of Scientific and Technical Information of China (English)

    YAO Bing; HU Xiaoli; BAO Zhenmin; LU Wei; HU Jingjie

    2007-01-01

    Sea cucumber Apostichopus japonicus samples were collected in Changdao, Penglai (PL),27 individuals, and Lingshandao, Qingdao (QD), 30 individuals, in the Shandong Peninsula, China. Ten SSR primers were used to assess the genetic variation and relationship between and within the two stocks.Respectively, for each stock, the percentage of polymorphic bands was 85.2% and 86.9%; the gene diversity was 0.360 5 and 0.342 8; and the Shannon's information index was 0.515 0 and 0.499 0. At species level, the percentage of polymorphic bands was 92.2%, the total gene diversity was 0.378 9 and the Shannon's information index was 0.550 4. The coefficient of overall genetic differentiation and the genetic distances between the stocks were also calculated to be 0.073 0 and 0.079 6 using the POPGENE program. Results show that the genetic diversity of the two stocks is still large but the genetic distance between the two stocks is close. A dendrogram was constructed for the 57 individuals from the two stocks,showing that the genetic structure was unitary for PL stock but complex for QD stock.

  9. Determination of genetic relationships among elite thermosensitive genic male sterile lines (TGMS) of rice (Oryza sativa L.) employing morphological and simple sequence repeat (SSR) markers

    Indian Academy of Sciences (India)

    Vikas Kumar Singh; Priti Upadhyay; Pallavi Sinha; Ashish Kumar Mall; Sanjay Kumar Jaiswal; Atul Singh; Ranjith Kumar Ellur; Sunil Biradar; R. M. Sundaram; Sukhpal Singh; Ilyas Ahmed; B. Mishra; A. K. Singh; C. Kole

    2011-04-01

    A set of morphological traits and SSR markers were used to determine the genetic relationship among 12 elite thermosensitive genic male sterile (TGMS) lines developed at three different research institutions of India. Agro-morphological data recorded on 20 morphological traits revealed a wide base of genetic variation and a set of four morphological traits could distinguish most of the TGMS lines. Analysis with 30 SSR markers (20 EST-SSRs and 10 genomic SSRs) revealed 27 markers to be polymorphic, amplifying a total of 83 alleles. Each SSR marker amplified 2–6 alleles with an average of 2.76 alleles per marker and a PIC value varying from 0.54 to 0.96. Cluster analysis based on SSR and morphological data clearly differentiated the lines according to their source of origin. Correlation analysis between morphological and molecular data revealed a very poor association ($r = 0.06$), which could be attributed to selection pressure, genetic drift, sampling error and unknown relationship among related lines. The SSR markers discriminated the genotypes distinctly and quantified the genetic diversity precisely among the TGMS lines. Data on the yield per plant indicated that the genotypes grouping under a similar cluster showed same heterotic behaviour as compared to the genotypes from different clusters when crossed to similar pollinators.

  10. Influence of single and repeated cannabidiol administration on emotional behavior and markers of cell proliferation and neurogenesis in non-stressed mice.

    Science.gov (United States)

    Schiavon, Angélica Pupin; Bonato, Jéssica Mendes; Milani, Humberto; Guimarães, Francisco Silveira; Weffort de Oliveira, Rúbia Maria

    2016-01-04

    Therapeutic effects of antidepressants and atypical antipsychotics may arise partially from their ability to stimulate neurogenesis. Cannabidiol (CBD), a phytocannabinoid present in Cannabis sativa, presents anxiolytic- and antipsychotic-like effects in preclinical and clinical settings. Anxiolytic-like effects of repeated CBD were shown in chronically stressed animals and these effects were parallel with increased hippocampal neurogenesis. However, antidepressant-like effects of repeated CBD administration in non-stressed animals have been scarcely reported. Here we investigated the behavioral consequences of single or repeated CBD administration in non-stressed animals. We also determined the effects of CBD on cell proliferation and neurogenesis in the dentate gyrus (DG) and subventricular zone (SVZ). Single CBD 3mg/kg administration resulted in anxiolytic-like effect in mice submitted to the elevated plus maze (EPM). In the tail suspension test (TST), single or repeated CBD administration reduced immobility time, an effect that was comparable to those of imipramine (20 mg/kg). Moreover, repeated CBD administration at a lower dose (3 mg/kg) increased cell proliferation and neurogenesis, as seen by an increased number of Ki-67-, BrdU- and doublecortin (DCX)-positive cells in both in DG and SVZ. Despite its antidepressant-like effects in the TST, repeated CBD administration at a higher dose (30 mg/kg) decreased cell proliferation and neurogenesis in the hippocampal DG and SVZ. Our findings show a dissociation between behavioral and proliferative effects of repeated CBD and suggest that the antidepressant-like effects of CBD may occur independently of adult neurogenesis in non-stressed Swiss mice.

  11. 利用Taguchi方法优化荆芥ISSR-PCR反应体系

    Institute of Scientific and Technical Information of China (English)

    王跃虎; 魏建和; 张东向; 隋春; 杨成民

    2007-01-01

    利用Taguchi方法对荆芥ISSR-PCR反应体系的5因素(dNTP,引物,模板DNA,Mg2+,Taq酶)在4个水平上进行优化试验,使用Quantity One4.6.2软件对电泳图片进行处理,PCR结果用SAS 9.1.3 SP4软件进行统计分析。得到如下主要结果:荆芥ISSR-PCR最佳反应体系(25μL):dNTP 169.0μmol/L,引物0.2μmol/L,模板DNA 20.6 ng,Mg2+浓度2.1 mmol/L,Taq酶1.2U;将目的条带权重赋值(s)引入PCR电泳结果量化赋值公式,增强其通用性;使用Taguchi方法进行荆芥ISSR-PCR反应体系优化效果明显,但建立的引物浓度回归方程无最大值,说明该方法的应用仍具有一定局限性。

  12. 不同放牧强度下冰草自然居群的遗传多样性分析%Study on ISSR of Agroptron cristatum at different grazing gradients

    Institute of Scientific and Technical Information of China (English)

    秀花

    2015-01-01

    The genetic diversity of Agropyron cristatum under different grazing intensity was studied with ISSR molecular marker technology.The results showed that although some polymorphic loci were missed due to grazing,the whole wheatgrass population still exhibited the rich polymorphism.Polymorphic bands ratio de-tected with ISSR was 96.46%,the genetic differences among populations was small,grazing did not produce genetic differentiation.Genetic diversity of each prime in 4 wheatgrass populations estimated by Shannon’s and Nei’s diversity index was:moderate grazing>severe grazing>banning grazing>light grazing.%采用ISSR分子标记技术对不同放牧强度下冰草自然居群的遗传多样性进行了研究。结果表明,放牧导致冰草部分位点丢失,但整个冰草居群仍表现出丰富的多态性,ISSR 检测的多态性条带比率为96.46%,居群间的遗传差异很小,放牧并未使冰草居群产生遗传分化。由 Shannon’s 和 Nei’多样性指数检测的各个引物在4个冰草居群内的遗传多样性的大小排列顺序为:中度放牧样地>重度放牧样地>无放牧样地>轻度放牧样地。

  13. Genetic diversity of Pinus nigra Arn. populations in Southern Spain and Northern Morocco revealed by inter-simple sequence repeat profiles.

    Science.gov (United States)

    Rubio-Moraga, Angela; Candel-Perez, David; Lucas-Borja, Manuel E; Tiscar, Pedro A; Viñegla, Benjamin; Linares, Juan C; Gómez-Gómez, Lourdes; Ahrazem, Oussama

    2012-01-01

    Eight Pinus nigra Arn. populations from Southern Spain and Northern Morocco were examined using inter-simple sequence repeat markers to characterize the genetic variability amongst populations. Pair-wise population genetic distance ranged from 0.031 to 0.283, with a mean of 0.150 between populations. The highest inter-population average distance was between PaCU from Cuenca and YeCA from Cazorla, while the lowest distance was between TaMO from Morocco and MA Sierra Mágina populations. Analysis of molecular variance (AMOVA) and Nei's genetic diversity analyses revealed higher genetic variation within the same population than among different populations. Genetic differentiation (Gst) was 0.233. Cuenca showed the highest Nei's genetic diversity followed by the Moroccan region, Sierra Mágina, and Cazorla region. However, clustering of populations was not in accordance with their geographical locations. Principal component analysis showed the presence of two major groups-Group 1 contained all populations from Cuenca while Group 2 contained populations from Cazorla, Sierra Mágina and Morocco-while Bayesian analysis revealed the presence of three clusters. The low genetic diversity observed in PaCU and YeCA is probably a consequence of inappropriate management since no estimation of genetic variability was performed before the silvicultural treatments. Data indicates that the inter-simple sequence repeat (ISSR) method is sufficiently informative and powerful to assess genetic variability among populations of P. nigra.

  14. Repeated measurements of blood lactate concentration as a prognostic marker in horses with acute colitis evaluated with classification and regression trees (CART) and random forest analysis.

    Science.gov (United States)

    Petersen, M B; Tolver, A; Husted, L; Tølbøll, T H; Pihl, T H

    2016-07-01

    The objective of this study was to investigate the prognostic value of single and repeated measurements of blood l-lactate (Lac) and ionised calcium (iCa) concentrations, packed cell volume (PCV) and plasma total protein (TP) concentration in horses with acute colitis. A total of 66 adult horses admitted with acute colitis (2 mmol/L (sensitivity, 0.72; specificity, 0.8). In conclusion, blood lactate concentration measured at admission and repeated 6 h later aided the prognostic evaluation of horses with acute colitis in this population with a very high mortality rate. This should allow clinicians to give a more reliable prognosis for the horse.

  15. Diversity of black Aspergilli isolated from raisins in Argentina: Polyphasic approach to species identification and development of SCAR markers for Aspergillus ibericus.

    Science.gov (United States)

    Giaj Merlera, G; Muñoz, S; Coelho, I; Cavaglieri, L R; Torres, A M; Reynoso, M M

    2015-10-01

    Aspergillus section Nigri is a heterogeneous fungal group including some ochratoxin A producer species that usually contaminate raisins. The section contains the Series Carbonaria which includes the toxigenic species Aspergillus carbonarius and nontoxigenic Aspergillus ibericus that are phenotypically undistinguishable. The aim of this study was to examine the diversity of black aspergilli isolated from raisins and to develop a specific genetic marker to distinguish A. ibericus from A. carbonarius. The species most frequently found in raisins in this study were Aspergillus tubingensis (35.4%) and A. carbonarius (32.3%), followed by Aspergillus luchuensis (10.7%), Aspergillus japonicus (7.7%), Aspergillus niger (6.2%), Aspergillus welwitschiae (4.6%) and A. ibericus (3.1%). Based on inter-simple sequence repeat (ISSR) fingerprinting profiles of major Aspergillus section Nigri members, a sequence-characterized amplified region (SCAR) marker was identified. Primers were designed based on the conserved regions of the SCAR marker and were utilized in a PCR for simultaneous identification of A. carbonarius and A. ibericus. The detection level of the SCAR-PCR was found to be 0.01 ng of purified DNA. The present SCAR-PCR is rapid and less cumbersome than conventional identification techniques and could be a supplementary strategy and a reliable tool for high-throughput sample analysis.

  16. The Perceived School Climate in Invitational Schools in Hong Kong: Using the Chinese Version of the Inviting School Survey-Revised (ISS-R)

    Science.gov (United States)

    Ng, Carmen K. M.; Yuen, Mantak

    2011-01-01

    This article describes the use of the Chinese translation of the revised Inviting School Survey (ISS-R; Smith, 2005; Smith & Bernard, 2004) to measure the invitational climate of seven invitational secondary schools in Hong Kong. The five subscales of Chinese version of ISS-R were found to be valid and reliable in a sample of 706 Grade 11…

  17. Genetic diversity of wild Cymbidium goeringii (Orchidaceae)populations from Hubei based on Inter-simple sequence repeats analysis

    Institute of Scientific and Technical Information of China (English)

    YAO Xiaohong; GAO Li; YANG Bo

    2007-01-01

    Cymbidium goeringii is a diploid and nonrewarding,bumblebee-pollinated species,which is distributed in China,Japan and Korea Peninsula.This species is now highly endangered due to the mass collection and forest clearance in China.In the present study,we investigated the distribution of genetic variation within and between eleven populations of Cymbidium goeringii in central China by using Inter-simple sequence repeats (ISSR) markers.Eleven primers produced a total of 127 clear and reproducible bands of which 112 were polymorphic.High genetic diversity was detected in Cymbidium goeringii for both population level (P = 63.1%;He = 0.194 5) and species level (P = 88.2%;He = 0.262 8).A higher level of genetic differentiation was detected among populations (GST = 0.244 0,FST = 0.220 7)with Nei's Gsr analysis and analysis of molecular variance (AMOVA),and no correlation was found between geographical and genetic distance.Genetic drift rather than gene flow played an important role in forming the present population structure of Cymbidium goeringii.Limited gene flow among populations and gene drift increase the extinction risk of local populations.Some conservation concerns are therefore discussed together with possible strategies for implementing in situ and ex situ conservation.

  18. Variable number of tandem repeat markers in the genome sequence of Mycosphaerella fijiensis, the causal agent of black leaf streak disease of banana (Musa spp)

    NARCIS (Netherlands)

    Garcia, S.A.L.; Lee, van der T.A.J.; Ferreira, C.F.; Lintel Hekkert, te B.; Zapater, M.F.; Goodwin, S.B.; Guzmán, M.; Kema, G.H.J.; Souza, M.T.

    2010-01-01

    ABSTRACT. We searched the genome of Mycosphaerella fijiensis for molecular markers that would allow population genetics analysis of this plant pathogen. M. fijiensis, the causal agent of banana leaf streak disease, also known as black Sigatoka, is the most devastating pathogen attacking bananas

  19. 适用于川芎的ISSR-PCR反应体系的建立及优化(英文)%Optimization of ISSR-PCR Reaction Conditions for Genomic DNA of Ligusticum chuanxiong hort.

    Institute of Scientific and Technical Information of China (English)

    王岚; 唐琳

    2012-01-01

    [Objective] To investigate the impacts of ISSR-PCR amplification factors, for the establishment and optimization of ISSR-PCR reaction system for Ligusticum chuanxiong hort. [Method] Using genomic DNA of Chuanxiong leaf extracted via an improved CTAB method as template, single factor analysis was performed to investigate the impacts of DNA template concentration, Mg2+ concentration, dNTPs concentration, primer concentration, Taq DNA polymerase concentration on ISSR-PCR amplification and to optimize this system for Ligusticum chuanxiong hort. [Result] The ISSR-PCR amplification(25 μl) suitable for Ligusticum chuanxiong hort. was determined to be composed of 2.5 μl of 10×reaction buffer, 2.1 mmol/L MgCl2, 300 μmol/L dNTPs, 0.4 μmol/L primer, 1.0 U Taq DNA polymerase and 20-40 ng genomic DNA. [Conclusion] Our study laid basis for analyzing the genetic diversity of Ligusticum chuanxiong hort. resources distributed in 17 different areas of China.%[目的]建立和优化适合川芎的ISSR-PCR反应体系。[方法]以川芎叶片为材料,利用改良CTAB法提取了叶片基因组DNA,利用单因素试验分析了DNA模板、Mg2+、dNTP、引物和TaqDNA聚合酶的浓度对ISSR-PCR反应的影响,对适合川芎的ISSR-PCR反应条件进行了优化。[结果]确定了适合川芎的ISSR-PCR反应体系为25.0μl,其中含2.5μl10×TaqDNA聚合酶缓冲液、2.1mmol/LMgCl2、300μmol/LdNTP、0.4μmol/L引物、1.0UTaqDNA聚合酶和20~40ng模板DNA。[结论]为进一步对全国17个不同分布区的川芎资源进行遗传多样性研究奠定了基础。

  20. Confamiliar transferability of simple sequence repeat (SSR) markers from cotton (Gossypium hirsutum L.) and jute (Corchorus olitorius L.) to twenty two Malvaceous species.

    Science.gov (United States)

    Satya, Pratik; Paswan, Pramod Kumar; Ghosh, Swagata; Majumdar, Snehalata; Ali, Nasim

    2016-06-01

    Cross-species transferability is a quick and economic method to enrich SSR database, particularly for minor crops where little genomic information is available. However, transferability of SSR markers varies greatly between species, genera and families of plant species. We assessed confamiliar transferability of SSR markers from cotton (Gossypium hirsutum) and jute (Corchorus olitorius) to 22 species distributed in different taxonomic groups of Malvaceae. All the species selected were potential industrial crop species having little or no genomic resources or SSR database. Of the 14 cotton SSR loci tested, 13 (92.86 %) amplified in G. arboreum and 71.43 % exhibited cross-genera transferability. Nine out of 11 jute SSRs (81.81 %) showed cross-transferability across genera. SSRs from both the species exhibited high polymorphism and resolving power in other species. The correlation between transferability of cotton and jute SSRs were highly significant (r = 0.813). The difference in transferability among species was also significant for both the marker groups. High transferability was observed at genus, tribe and subfamily level. At tribe level, transferability of jute SSRs (41.04 %) was higher than that of cotton SSRs (33.74 %). The tribe Byttnerieae exhibited highest SSR transferability (48.7 %). The high level of cross-genera transferability (>50 %) in ten species of Malvaceae, where no SSR resource is available, calls for large scale transferability testing from the enriched SSR databases of cotton and jute.

  1. Phylogenetic analysis, genetic diversity and relationships between the recently segregated species of Corynandra and Cleoserrata from the genus Cleome using DNA barcoding and molecular markers.

    Science.gov (United States)

    Tamboli, Asif Shabodin; Patil, Swapnil Mahadeo; Gholave, Avinash Ramchandra; Kadam, Suhas Kishor; Kotibhaskar, Shreya Vijaykumar; Yadav, Shrirang Ramchandra; Govindwar, Sanjay Prabhu

    2016-01-01

    Cleome is the largest genus in the family Cleomaceae and it is known for its various medicinal properties. Recently, some species from the Cleome genus (Cleome viscosa, Cleome chelidonii, Cleome felina and Cleome speciosa) are split into genera Corynandra (Corynandra viscosa, Corynandra chelidonii, Corynandra felina), and Cleoserrata (Cleoserrata speciosa). The objective of this study was to obtain DNA barcodes for these species for their accurate identification and determining phylogenetic relationships. Out of 10 screened barcoding regions, rbcL, matK and ITS1 regions showed higher PCR efficiency and sequencing success. This study added matK, rbcL and ITS1 barcodes for the identification of Corynandra chelidonii, Corynandra felina, Cleome simplicifolia and Cleome aspera species in existing barcode data. Corynandra chelidonii and Corynandra felina species belong to the Corynandra genus, but they are not grouped with the Corynandra viscosa species, however clustered with the Cleome species. Molecular marker analysis showed 100% polymorphism among the studied plant samples. Diversity indices for molecular markers were ranged from He=0.1115-0.1714 and I=0.2268-0.2700, which indicates a significant amount of genetic diversity among studied species. Discrimination of the Cleome and Corynandra species from Cleoserrata speciosa was obtained by two RAPD primers (OPA-4 and RAPD-17) and two ISSR primers (ISSR-1 and ISSR-2). RAPD and ISSR markers are useful for the genetic characterization of these studied species. The present investigation will be helpful to understand the relationships of Cleome lineages with Corynandra and Cleoserrata species.

  2. Assessment of genetic diversity in a highly valuable medicinal plant Catharanthus roseus using molecular markers

    Directory of Open Access Journals (Sweden)

    Ranjan Kumar Shaw

    2009-01-01

    Full Text Available Genetic diversity was evaluated among 14 cultivars of Catharanthus roseus using RAPD and ISSR markers.The RAPD primers resulted in the amplification of 56 bands, among which 46 (82% bands were polymorphic Four ISSRprimers amplified 31 loci out of which 17 were polymorphic and 14 are monomorphic. The Jaccard's similarity derived fromthe combined marker system showed that the varieties First Kiss Coral and Cooler Orchid were the most closely relatedcultivars, with 98% similarity. In the dendrogram constructed on the basis of both RAPD and ISSR data two clear clusterswere obtained. The smaller cluster included C. roseus Cv Blue Pearl and C. roseus Cv. Patricia White and the larger clusterwas subdivided into two sub clusters with C. roseus Cv. First Kiss Polka Dot isolated from the rest of the cultivars. This maybe useful for breeding for improved quality.

  3. Unraveling the evolutionary scenario of the hobo element in populations of Drosophila melanogaster and D. simulans in South America using the TPE repeats as markers

    Directory of Open Access Journals (Sweden)

    Geovani T. Ragagnin

    2016-03-01

    Full Text Available Abstract Transposable elements (TEs are nucleotide sequences found in most studied genomes. These elements are highly diversified and have a large variation in nucleotide structure and mechanisms of transposition. hobo is a member of class II, belonging to hAT superfamily, described inDrosophila melanogaster, and it presents in its Open Reading Frame, a repetitive region encoding the amino acids threonine-proline-glutamic acid (TPE, which shows variability in the number of repeats in some regions of the world. Due to this variability some evolutionary scenarios of the hobo element are discussed, such as the scenario of the invasion of hobo element in populations ofD. melanogaster. In the present study, we investigated 22 DNA sequences of D. melanogaster and seven sequences ofD. simulans, both from South America, to check the number of repetitions of TPE, in order to clarify the evolutionary scenario of thehobo element in these populations. Our results showed a monomorphism in populations of both species in South America, with only three TPE repeats. Hence, we discuss and propose an evolutionary scenario of the invasion of the hobo element in populations of D. melanogaster and D. simulans.

  4. Identification of a glutamic acid repeat polymorphism of ALMS1 as a novel genetic risk marker for early-onset myocardial infarction by genome-wide linkage analysis.

    Science.gov (United States)

    Ichihara, Sahoko; Yamamoto, Ken; Asano, Hiroyuki; Nakatochi, Masahiro; Sukegawa, Mayo; Ichihara, Gaku; Izawa, Hideo; Hirashiki, Akihiro; Takatsu, Fumimaro; Umeda, Hisashi; Iwase, Mitsunori; Inagaki, Haruo; Hirayama, Haruo; Sone, Takahito; Nishigaki, Kazuhiko; Minatoguchi, Shinya; Cho, Myeong-Chan; Jang, Yangsoo; Kim, Hyo-Soo; Park, Jeong E; Tada-Oikawa, Saeko; Kitajima, Hidetoshi; Matsubara, Tatsuaki; Sunagawa, Kenji; Shimokawa, Hiroaki; Kimura, Akinori; Lee, Jong-Young; Murohara, Toyoaki; Inoue, Ituro; Yokota, Mitsuhiro

    2013-12-01

    Myocardial infarction (MI) is a leading cause of death worldwide. Given that a family history is an independent risk factor for coronary artery disease, genetic variants are thought to contribute directly to the development of this condition. The identification of susceptibility genes for coronary artery disease or MI may thus help to identify high-risk individuals and offer the opportunity for disease prevention. We designed a 5-step protocol, consisting of a genome-wide linkage study followed by association analysis, to identify novel genetic variants that confer susceptibility to coronary artery disease or MI. A genome-wide affected sib-pair linkage study with 221 Japanese families with coronary artery disease yielded a statistically significant logarithm of the odds score of 3.44 for chromosome 2p13 and MI. Further association analysis implicated Alström syndrome 1 gene (ALMS1) as a candidate gene within the linkage region. Validation association analysis revealed that representative single-nucleotide polymorphisms of the ALMS1 promoter region were significantly associated with early-onset MI in both Japanese and Korean populations. Moreover, direct sequencing of the ALMS1 coding region identified a glutamic acid repeat polymorphism in exon 1, which was subsequently found to be associated with early-onset MI. The glutamic acid repeat polymorphism of ALMS1 identified in the present study may provide insight into the pathogenesis of early-onset MI.

  5. ISSR Analysis of M_1 Generation of Gladiolus hybridus Hort Treated by EMS

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Gladiolus hybridus Hort is one of famous cutting flowers,being famous of big size,bright color,various shapes and long bloom period.New species should be cultivated in order to meet consumers' needs.Mutagenic breeding is a shortcut to cultivate new species of flowers.In this study,corm bud of G.hybridus Hort was treated with different concentrations of EMS.Then M 1 generation was analyzed by ISSR.Results showed that EMS was a very effective mutagenic agent for the corm bud of G.hybridus Hort.With the increa...

  6. Isolation and Characterization of Simple Sequence Repeats (SSR Markers from the Moss Genus Orthotrichum Using a Small Throughput Pyrosequencing Machine

    Directory of Open Access Journals (Sweden)

    Vítězslav Plášek

    2012-06-01

    Full Text Available Here, we report the results of next-generation sequencing on the GS Junior system to identify a large number of microsatellites from the epiphytic moss Orthotrichum speciosum. Using a combination of a total (non-enrichment genomic library and small-scale 454 pyrosequencing, we determined 5382 contigs whose length ranged from 103 to 5445 bp. In this dataset we identified 92 SSR (simple sequence repeats motifs in 89 contigs. Forty-six of these had flanking regions suitable for primer design. We tested PCR amplification, reproducibility, and the level of polymorphism of 46 primer pairs for Orthotrichum speciosum using 40 individuals from two populations. As a result, the designed primers revealed 35 polymorphic loci with more than two alleles detected. This method is cost- and time-effective in comparison with traditional approaches involving cloning and sequencing.

  7. 甜瓜属REMAP分子标记体系的建立及应用%Development and Application of REMAP Molecular Marker System in Cucumis

    Institute of Scientific and Technical Information of China (English)

    王东; 江彪; 娄群峰; 陈劲枫

    2011-01-01

    REMAP (retrotransposon-microsatellite amplified polymorphism) marker system was developed based on the long terminal repeat(LTR) sequence of Tyl-copia retrotransposon in Cucumis and combination of the ISSR primer sequence.Forty-six cultigens in Cucum/s were genotyped using REMAP markers and the results indicated REMAP can efficiently detect polymorphism among different melon material. Thus it can be used for cultivar identification and fingerprinting in Cucumis.%基于甜瓜属Ty1-copia类逆转座子的长末端重复序列信息,结合ISSR引物序列,在体系优化的基础上建立了适用于甜瓜属物种的REMAP(retrotransposon-microsatellite amplified polymorphism)标记体系.对46份甜瓜属不同类型材料进行聚类分析,证明该标记体系能有效检测甜瓜属不同类型材料间的多态性,可以用于甜瓜属作物品种鉴定及指纹图谱构建.

  8. 适用于川芎的ISSR-PCR反应体系的建立及优化%Establishment and Optimization of ISSR-PCR Amplification System for Ligusticum chuanxiong Hort

    Institute of Scientific and Technical Information of China (English)

    王岚; 唐琳

    2011-01-01

    [Objective] This study was to establish and optimize the ISSR-PCR reaction system for Ligusticum chuanxiong Hort. [ Method] Using genomic DNA of Chuanxiong leaf extracted via an improved CTAB method, single factor analysis was performed to investigate the impacts of DNA template concentration, Mg2+ concentration, dNTPs concentration, primer concentration, Taq DNA polymerase concentration on ISSR-PCR amplification and to optimize this system for Ligusticum chuanxiong Hort. [ Result] The ISSR-PCR amplification(25 μl) suitable for Ligusticum chuanxiong Hort. Was determined to be composed of 2. 5 μl of 10 × reaction buffer, 2.1 mmol/L MgCl2, 300 μmol/L dNTPs, 0.4 μmol/L primer, 1.0 U Taq DNA polymerase and 20 -40 ng genomic DNA. [ Conclusion] Our study laid basis for analyzing the genetic diversity of Ligusticum chuanxiong Hort. Resources distributed in 17 different areas of China.%[目的]建立和优化适合川芎的ISSR-PCR反应体系.[方法]以川芎叶片为材料,利用改良CTAB法提取了叶片基因组DNA,利用单因素试验分析了DNA模板、Mg2+、dNTP、引物和Taq DNA聚合酶的浓度对ISSR-PCR反应的影响,对适合川芎的ISSR-PCR反应条件进行了优化.[结果]适合川芎的ISSR-PCR反应体系为25.0μl,其中含2.5μl10×Taq DNA聚合酶缓冲液、2.1 mmol/L MgCl2、300μmol/L dNTP、0.4 μmol/L引物、1.0 U Taq DNA聚合酶和20 ~40 ng模板DNA.[结论]为进一步对全国17个不同分布区的川芎资源进行遗传多样性研究奠定了基础.

  9. ISSR and RAPD based evaluation of genetic stability of encapsulated micro shoots of Glycyrrhiza glabra following 6 months of storage.

    Science.gov (United States)

    Mehrotra, Shakti; Khwaja, O; Kukreja, A K; Rahman, L

    2012-11-01

    In vitro grown axillary micro shoots of Glycyrrhiza glabra were encapsulated in alginate beads. Following 6 months of normal storage at 25 ± 2°C the re growth of encapsulated G. glabra micro shoots, reached 98% within 30 days of incubation on MS medium supplemented with 0.1 mg/l IAA. Re growth was characterized by the development of both shoot and root from single encapsulated micro shoot. Healthy plants were established to glass house with 95% survival. The genetic fidelity of plants obtained after conversion of alginate beads was ascertained through 10 RAPD and 13 ISSR primers. Of the 10 RAPD primers tested, 6 of them produced 14 clear and reproducible amplicons with an average of 2.3 bands per primer out of which 28.57% were polymorphic generated by only two primers. Eight ISSR primers produced total 37 bands ranging between 300 and 3,500 bp length. Number of scorable bands for each primer varied from 3 to 8 with an average of 4.6 bands per primer. Cluster analysis from ISSR and RAPD showed that all the tested plants including the mother plant distributed in two major groups with similarity coefficient ranging from 0.91 to 0.96 for RAPD and 0.89 to 0.97 for ISSR.

  10. Establishment and Optimization of ISSR-PCR Reaction System for Male Sterile Lines of Marigold%万寿菊雄性不育系ISSR-PCR体系建立与优化

    Institute of Scientific and Technical Information of China (English)

    伍亚平; 唐道城

    2013-01-01

    The experimental designs with a single-factor and orthogonal test was used to optimize the ISSR-PCR reaction procedures of nine male sterile lines in marigold in four factors including the concentration of Taq DNA polymerase, dNTP, Primers and Mg2+. Then, the template concentration and ISSR primers were screened and validated based on the optimized ISSR-PCR system, the annealing temperature of the primers were detected by gradient PCR approach as well. The optimal concentration of each factors for ISSR-PCR amplification system of marigold male sterile lines were determined as Mg2+1.5 mmol/L, dNTPs 0.20 mmol/L, primer 0.5μmol/L, template DNA 60 ng and Taq DNA polymerase 1 U in reaction volume of 25μL.%  以9个万寿菊雄性不育系总DNA为材料,利用单因素试验和正交试验设计对Mg2+、dNTPs、引物和Taq酶的浓度进行筛选,然后在此基础上筛选模板DNA浓度及ISSR引物并对引物退火温度进行梯度检测,优化得到最佳的ISSR-PCR反应体系,各个因素的最佳水平为:在25μL反应体系中,Mg2+浓度为1.5 mmol/L、dNTPs浓度为0.20 mmol/L、引物浓度为0.5μmol/L、模板DNA为60 ng、Taq DNA聚合酶含量为1 U.

  11. Comparison of ITS, RAPD and ISSR from DNA-based genetic diversity techniques.

    Science.gov (United States)

    Poyraz, Ismail

    2016-01-01

    ITS, RAPD-PCR and ISSR-PCR are most popular DNA-based techniques that are extensively applied in the determination of the genetic diversity of species among populations. However, especially for organisms having high genetic polymorphism, phylogenetic trees drawn from the results of these techniques may be different. For finding a meaningful phylogenetic tree, it should be compared phylogenetic trees obtained from these different techniques with geographic locations of populations. Lichens have a high genetic polymorphism and tolerance against different environmental conditions. In this study, these three DNA-based genetic diversity techniques were compared, using different populations of a lichen species (Xanthoria parietina). X. parietina was especially chosen because of its high genetic diversity in narrow zones. Lichen samples were collected from ten different locations in a narrow transition climate zone Bilecik (Turkey). Statistical analyses of all results were calculated using UPGMA analysis. Phylogenic trees for each technique were drawn and transferred to the Bilecik map for comparative analysis. The results of three techniques allowed us to verify that populations of X. parietina have high genetic variety in a narrow zone. But phylogenetic trees obtained from these results were found to be very different. Our comparative analysis demonstrated that the results of these techniques are not similar and have critical differences. We observed that the ITS method provides more clear data and is more successful in genetic diversity analyses of more asunder populations, in contrast to ISSR-PCR and RAPD-PCR methods.

  12. Repeatedly Heading a Soccer Ball Does Not Increase Serum Levels of S-100B, a Biochemical Marker of Brain Tissue Damage: an Experimental Study.

    Science.gov (United States)

    Stålnacke, Britt-Marie; Sojka, Peter

    2008-02-29

    OBJECTIVES: The aim of the study was to analyse whether the controlled heading of soccer balls elicits increased serum concentrations of a biochemical marker of brain tissue damage S-100B. METHODS: Nineteen male soccer players were randomly divided into two groups, A and B. Group A headed a soccer ball falling from 18 m five times, while group B served as controls (no heading). Blood samples were taken before and 0.5 h, 2 h and 4 h after the heading for analysis of S-100B. RESULTS: No statistically significant (p > 0.05) increases in serum concentrations of S-100B were encountered in group A at 0.5 h (0.109 +/-0.024 mug/L), 2 h (0.098 +/- 0.026 mug/L), and 4 h (0.113 +/- 0.035 mug/L) when the blood samples obtained before and after the heading were compared (0.157 +/- 0.134 mug/L). No statistically significant difference was found when the serum concentrations of S-100B were compared between groups A and B either before or after heading. CONCLUSIONS: Heading a soccer ball dropped from a height of 18 m five times was not found to cause an increase in serum concentrations of S-100B, indicating that the impact was not sufficient to cause biochemically discernible damage of brain tissue.

  13. Repeatedly Heading a Soccer Ball Does Not Increase Serum Levels of S-100B, a Biochemical Marker of Brain Tissue Damage: an Experimental Study

    Directory of Open Access Journals (Sweden)

    Peter Sojka

    2008-01-01

    Full Text Available Objectives: The aim of the study was to analyse whether the controlled heading of soccer balls elicits increased serum concentrations of a biochemical marker of brain tissue damage S-100B.Methods: Nineteen male soccer players were randomly divided into two groups, A and B. Group A headed a soccer ball falling from 18 m five times, while group B served as controls (no heading. Blood samples were taken before and 0.5 h, 2 h and 4 h after the heading for analysis of S-100B.Results: No statistically significant (p > 0.05 increases in serum concentrations of S-100B were encountered in group A at 0.5 h (0.109 ± 0.024 μg/L, 2 h (0.098 ± 0.026 μg/L, and 4 h (0.113 ± 0.035 μg/L when the blood samples obtained before and after the heading were compared (0.157 ± 0.134 μg/L. No statistically significant difference was found when the serum concentrations of S-100B were compared between groups A and B either before or after heading.Conclusions: Heading a soccer ball dropped from a height of 18 m five times was not found to cause an increase in serum concentrations of S-100B, indicating that the impact was not sufficient to cause biochemically discernible damage of brain tissue.

  14. Comparison of Variable-Number Tandem-Repeat Markers typing and IS1245 Restriction Fragment Length Polymorphism fingerprinting of Mycobacterium avium subsp. hominissuis from human and porcine origins

    Directory of Open Access Journals (Sweden)

    Marttila Harri

    2010-03-01

    Full Text Available Abstract Background Animal mycobacterioses are regarded as a potential zoonotic risk and cause economical losses world wide. M. avium subsp. hominissuis is a slow-growing subspecies found in mycobacterial infected humans and pigs and therefore rapid and discriminatory typing methods are needed for epidemiological studies. The genetic similarity of M. avium subsp. hominissuis from human and porcine origins using two different typing methods have not been studied earlier. The objective of this study was to compare the IS1245 RFLP pattern and MIRU-VNTR typing to study the genetic relatedness of M. avium strains isolated from slaughter pigs and humans in Finland with regard to public health aspects. Methods A novel PCR-based genotyping method, variable number tandem repeat (VNTR typing of eight mycobacterial interspersed repetitive units (MIRUs, was evaluated for its ability to characterize Finnish Mycobacterium avium subsp. hominissuis strains isolated from pigs (n = 16 and humans (n = 13 and the results were compared with those obtained by the conventional IS1245 RFLP method. Results The MIRU-VNTR results showed a discriminatory index (DI of 0,92 and the IS1245 RFLP resulted in DI 0,98. The combined DI for both methods was 0,98. The MIRU-VNTR test has the advantages of being simple, reproducible, non-subjective, which makes it suitable for large-scale screening of M. avium strains. Conclusions Both typing methods demonstrated a high degree of similarity between the strains of human and porcine origin. The parallel application of the methods adds epidemiological value to the comparison of the strains and their origins. The present approach and results support the hypothesis that there is a common source of M. avium subsp. hominissuis infection for pigs and humans or alternatively one species may be the infective source to the other.

  15. Prognostic significance of leucine-rich-repeat-containing G-protein-coupled receptor 5, an intestinal stem cell marker, in gastric carcinomas.

    Science.gov (United States)

    Jang, Bo Gun; Lee, Byung Lan; Kim, Woo Ho

    2016-07-01

    Cells expressing LGR5, an intestinal stem cell marker, have been suggested as cancer stem cells in human colon cancers. Previously, we discovered that LGR5-expressing cells exist in the gastric antrum and remarkably increase in number in intestinal metaplasia. In addition, most gastric adenomas contain abundant LGR5-expressing cells coexpressing intestinal stem cell signatures. However, LGR5 expression in gastric cancers (GCs) and its prognostic significance remain unknown. We examined the LGR5 expression in GC tissues by real time-PCR and RNA in situ hybridization, and analyzed its clinicopathological relevance and prognostic value. The effects of LGR5 on cancer cell proliferation and migration were assessed with an in vitro transfection technique. LGR5 expression was significantly lower in GCs than in matched nontumorous gastric mucosa. RNA in situ hybridization on tissue microarrays showed that 7 % of GCs were positive for LGR5. LGR5 positivity was associated with old age, well to moderate differentiation, and nuclear β-catenin positivity. Although LGR5 did not show any prognostic significance for all GC cases, it was associated with poor survival in GCs with nuclear β-catenin expression. LGR5 expression was induced by transfection in GC cell lines with abnormal Wnt activation, which, however, showed no influence on the growth and migration of GC cells. A small portion of GCs expressed LGR5. Although LGR5 was associated with poor survival in GCs with nuclear β-catenin, LGR5 expression in GC cells had no effects on the growth and migration, requiring a further study exploring a biological role of LGR5 in GCs.

  16. 5个温郁金居群遗传多样性的ISSR分析%The genetic diversity of the five populations Curcuma wenyujin by ISSR analysis

    Institute of Scientific and Technical Information of China (English)

    王佐元; 陶银龙; 郑蔚虹

    2013-01-01

    The genetic diversity of five populations of Curcuma wenyujin,a species endemic to Ruian,Yueqing Pingyang and,was analyzed using inter simple sequence repeat(ISSR). A total of 174 6 bands and 42 loci were amplified from 5 populations by 6 informative and reliable primers of 40 primers. Of them,34 loci were polymorphic, which accounted for 80.95%. Genetic diversity was revealed,Ht=0.239 5,Hs=0.1618,Dst=0.077 7 and GST=0.324 5. The study shows that there has a middle degree of genetic differentiation occurred among Curcuma wenyujin populations and a high genetic variation level beyond the populations of Curcuma wenyujin. Thus,Nm=1.041 0 means genetic was more keep in touch with each other among the population of Curcuma wenyujin.%应用ISSR标记技术对瑞安、乐清和平阳三个地区5个居群的温郁金(Curcuma wenyujin)遗传多样性进行分析,筛选出6个引物用于温郁金5个居群的50个个体ISSR-PCR扩增,共检测到42个结合位点多态位点34个,多态位点百分率是80.95%.总的基因多样度平均值为0.2395,居群内的基因多样度为0.1618,居群间的基因多样性为0.0777.基因分化系数在0.0191~0.9414之间,平均值为0.3245,其中居群间的遗传变异占总遗传变异的32.45%,居群内的遗传变异占总变异的67.55%.温郁金有着较高的遗传多样性水平,且居群内发生了较高的遗传分化.基因流系数Nm为1.0410,表明温郁金居群间的基因交流也比较多.

  17. 基于ISSR技术研究诺丽种质资源的亲缘关系%Genetic Relationship ofMorinda citrifolia Germplasms by ISSR

    Institute of Scientific and Technical Information of China (English)

    吴田; 蓝增全

    2014-01-01

    In order to understand the genetic relationship ofMorinda citrifolia(noni), the DNA fingerprints of 13 noni germplasm resources of were studied by ISSR markers. The results showed that ten ISSR primers could amplify 183 bands, of which 159 bands were polymorphic, accounting for 86.9%. The genetic similarity coefifcients of 13 noni germplasm resources ranged from 0.464 to 0.784. Cluster analysis showed that 13 noni germplasm resources could be divided into two clads, in which Small fruit as a single group had distant relation with other germplasm resources. Although all noni germplasm resources could not completely separated according external morphological characteristics, the most with the same characteristics could be clustered into the same group or sub-group.%为探讨我国诺丽(Morinda citrifolia)种质资源的亲缘关系,采用ISSR技术对诺丽种质资源的遗传关系进行分析。结果表明,10条ISSR引物对13份诺丽种质资源共扩增出183条带,其中多态性条带有159条,占86.9%。13份诺丽种质的遗传相似系数为0.464~0.784。聚类分析将13份诺丽种质资源聚为两类,其中诺丽小黑种单独聚为一类,与其他12份诺丽种质资源的亲缘关系较远。虽然按照外部形态特征不能将所有诺丽种质完全区分,但具有相同特征的多数种质还是聚在同一类或亚类中。

  18. 23个广西本地芒果品种的ISSR分析%Genetic Analysis of 23 Mango Cultivar Collection in Guangxi Province Revealed by ISSR

    Institute of Scientific and Technical Information of China (English)

    何新华; 李杨瑞; 郭永泽; 唐志鹏; 李容柏

    2005-01-01

    本研究对23个分别来自广西本地品种或广西选育的品种进行了SSR分析,并用15个芒果栽培品种印度(4个品种)、美国(3个品种)、缅甸(3个品种)、泰国(2个品种)及菲律宾、巴基斯坦和斯里兰卡(各1个品种)和2个芒果近源种冬芒(1个)和扁桃(1个)作为参照.从42个ISSR引物中筛选出9个多态性好的引物构建DNA指纹图谱来鉴别芒果的基因型.根据这9个引物获得的电泳图谱结果表明,本研究中所有供试品种能互相区分开来并表现出丰富的遗传多样性,说明ISSR是一种鉴定芒果品种(系)非常有效的方法.用74条多态性条带聚类分析作图发现所有供试的广西品种与秋芒、马切苏芒、象牙芒和爱文芒归为一个大类,而且广西本地芒果品种之间的亲缘关系较近.%Twenty-three mango cultivars (Mangifera indica Linn) collecting in Guangxi province were examined by ISSR markers and two species relatives M. himalis J.Y. Liang and M. persiciformis Wu & Ming, other 15mango cultivars from India (4), USA (3), Burma (3), Thailand (2), Philippines (1), Pakistan (1) and Sir Lanka (1)used as references. Of the 42 primers screened, 9 primers gave reproducible, polymorphic DNA amplification patterns, and were selected to construct a DNA fingerprinting map to distinguish the genotypes of mango. According to the banding patternsobtained with 9 selected primers, all cultivars tested in this study were distinguished from each other and showed ample genetic diversity, indicating that ISSR-PCR was an effective method for cultivar identification of mangoes. Based on 74 selected bands, all Guangxi mango cultivars tested were clustered into a big group with Neelum, Macheso, Aroemwnis, and Irwin by UPGMA analysis, indicating that Guangxi mango cultivars had a close relationship each other.

  19. White Kwao Krua variety classification by botanical characteristics and ISSR-Touchdown PCR technique.

    Science.gov (United States)

    Bunmanop, S; Sakuanrungsirikul, S; Manakasem, Y

    2011-07-01

    White Kwao Krua [Pueraria candollei Grah. var. mirifica (Airy Shaw et Suvatabandhu) Niyomdham] is a herb used as an ingredient in supplementary and cosmetic. The tuberous roots of White Kwao Krua (WKK) contain estrogen-like substances. Seeds of WKK, collected from Prachuab Khiri Khan, were planted and propagated in the farm of Suranaree University of Technology, and their genetic backgrounds were ambiguous. Thirty six plants of WKK in the same age were sampled for classification using 7 botanical characteristics and DNA fingerprint by ISSR-Touchdown PCR technique. The relationship of the 7 botanical characteristics, using principle component analysis (PCA), showed the WKK plants fell into 3 groups. In the first group was plant number 34, which was distinguished from the other plants by its small leaf size. The second group consisted of 23 plants with elliptic leaf shape, acute leaf base, and acuminate leaf apex. The third group consisted of 12 plants with ovate leaf shape, obtuse leaf base, and cuspidate leaf apex. The ISSR-Touchdown PCR technique with 41 primers detected 355 loci of DNA with an average of 8.6 loci per primer. The sizes of DNA ranged between 280 bp to 1550 bp. Two hundred ninety three loci exhibited polymorphisms (82.54%) and the rest 62 loci were monomorphic (17.46%). The polymorphism information content (PIC) was between 0.0315-0.9779 (average 0.4779) and number of effective alleles per locus (Ne) ranged between 1.1250-1.8541 (average 1.5544). Unweighted pair group method with arithmetic mean (UPGMA), Jaccard similarity coefficient and PCA were used to find the construction of genetic relationship of WKK. The genetic similarity (GS) of WKK ranged between 0.50-0.86 (average 0.77). At the GS of 0.56 from cluster analysis, the WKK varieties could be divided into 2 major groups. The first group comprised of plant number 34 and 7, and the second group could be further divided in 2 subgroups at GS of 0.69. None of the WKK plants was identical in

  20. Improvement of Genomic DNA Extraction Method and Optimization of ISSR-PCR Amplification for Citrus%柑橘基因组DNA快速提取及ISSR-PCR扩增体系优化

    Institute of Scientific and Technical Information of China (English)

    施维属; 潘腾飞; 钟凤林; 潘东明; 李开拓; 王江波; 郭志雄; 刘天亮

    2009-01-01

    对CTAB-mini法进行改良,得到一种效率较高的柑橘DNA提取方法.试验结果表明,得到的高质量DNA适合柑橘ISSR分析.同时,采用正交设计对影响柑橘ISSR-PCR因子进行优化.试验结果表明:20 μl反应体系中采用5 ng模板DNA、0.35 mmol/L dNTPs、0.15 μmol/L Primer、1U Taq酶和3 μl 10×Buffer(含Mg2+)为柑橘ISSR-PCR最优反应体系.

  1. Interactions of MinK and e-NOS gene polymorphisms appear to be inconsistent predictors of atrial fibrillation propensity, but long alleles of ESR1 promoter TA repeat may be a promising marker.

    Science.gov (United States)

    Smalcelj, Anton; Sertić, Jadranka; Golubić, Karlo; Jurcić, Ljiljana; Banfić, Ljiljana; Brida, Margarita

    2009-09-01

    Interactions of MinK and e-NOS Gene Polymorphisms Appear to Be Inconsistent Predictors of Atrial Fibrillation Propensity, but Long Alleles of ESR1 Promoter TA Repeat May Be a Promising Marker. We analyzed minK, e-NOS and ESR1 gene polymorphisms in 40 patients with atrial fibrillation (AF) without major structural heart disease compared to 35 healthy controls. A missense polymorphism in the minK gene with A/G substitution at nucleotide 112 causing serine (S) to glycine (G) change, 786 T/C polymorphism in the 5' flanking region of e-NOS gene and TA polymorphism in the regulatory region of estrogen receptor ESR1 gene with long (> or = 19 TA repeats) and short alleles were examined. Only a slight increase in minK G allele frequency, but with marked excess in AG/TT combination of minK and e-NOS polymorphisms was found in the AF group. The interpretation remains tentative due to small groups precluding statistical significance of differences, possible lab flaws and inconsistencies with earlier data. However, ESR1 long allele homozygotes were strikingly more frequent in the AF than in control group, reaching statistical significance surprisingly in males (p < 0.02). Functional activity of estrogen receptors may be more critical in males than in females with abundance of circulating estrogen. Contrasting the intricate complexity of genetic polymorphisms influencing cardiac rhythm with our modest research, we would limit the conclusion to the plea for further research of ESR1 role in AF.

  2. Two-step identification of taro (Colocasia esculenta cv. Xinmaoyu) using specific psbE-petL and simple sequence repeat-sequence characterized amplified regions (SSR-SCAR) markers.

    Science.gov (United States)

    Dai, H J; Zhang, Y M; Sun, X Q; Xue, J Y; Li, M M; Cao, M X; Shen, X L; Hang, Y Y

    2016-01-01

    Colocasia esculenta cv. Xinmaoyu is an eddoe-type taro cultivar local to Taicang, Jiangsu Province, China; it is characterized by its pure flavor, glutinous texture, and high nutritional value. Due to its excellent qualities, the Trademark Office of the State Administration for Industry and Commerce of the People's Republic of China awarded Xinmaoyu, a geographical indication certification in 2014. Therefore, there is an urgent need to develop an efficient molecular marker for the specific identification of this cultivar, which would greatly facilitate the conservation and utilization of this unique germplasm resource. In the present study, amplifying the psbE-petL fragment from two dasheen-type and seven eddoe-type taro cultivars revealed three conserved insertions/deletions among sequences from the two taro types. Based on these sequence differences, a pair of site-specific primers was designed targeting the psbE-petL sequence from the dasheen-type taro, which specifically amplified a DNA band in all individuals from cultivars of this type, but not in those from the seven eddoe-type cultivars. To discriminate Xinmaoyu from the other eddoe-type taro cultivars, a pair of simple sequence repeat-sequence characterized amplified region (SSR-SCAR) primers was further developed to specifically amplify a DNA band from all Xinmaoyu individuals, but not from individuals of other eddoe-type taro cultivars. In conclusion, through a two-step-screening procedure using psbE-petL and SSR-SCAR markers, we developed a pair of primers that could specifically discriminate Xinmaoyu from nine taro cultivars commonly cultivated in Jiangsu Province and Fujian Province.

  3. Establishment and Optimization of ISSR-PCR Reaction System for Crisp Melon (Cucumis melo var.conomon Group)%酥瓜(Cucumis melo var.conomon Group) ISSR-PCR反应体系建立与优化

    Institute of Scientific and Technical Information of China (English)

    葛继涛; 陈国户; 袁凌云; 朱世东; 苏亚; 刘思嘉; 方柔; 汪承刚

    2016-01-01

    利用正交设计L16(45),对酥瓜ISSR-PCR反应体系的5个影响因素(引物,dNTPs,Taq DNA聚合酶,Mg2+和模板DNA)在4个水平上进行优化试验,并在36℃~56℃范围内摸索退火温度,建立适合酥瓜ISSR-PCR反应体系,结果表明,在20μL反应体系中,含有引物0.2 μmol/L、dNTPs 0.3 mmol/L、Taq DNA聚合酶1.2 U、Mg2+ 1.0 mmol/L、DNA模板70 ng、10x Buffer 2.0μL为最佳反应体系,ISSR-PCR扩增程序中最佳退火温度为52.5℃.该体系为酥瓜种质资源的遗传多样性分析评价提供了帮助.

  4. Automated quality checks on repeat prescribing.

    OpenAIRE

    Rogers, Jeremy E; Wroe, Christopher J; Roberts, Angus; Swallow, Angela; Stables, David; Cantrill, Judith A; Rector, Alan L.

    2003-01-01

    BACKGROUND: Good clinical practice in primary care includes periodic review of repeat prescriptions. Markers of prescriptions that may need review have been described, but manually checking all repeat prescriptions against the markers would be impractical. AIM: To investigate the feasibility of computerising the application of repeat prescribing quality checks to electronic patient records in United Kingdom (UK) primary care. DESIGN OF STUDY: Software performance test against benchmark manual...

  5. Virulence Structure of Blumeria graminis f. sp. tritici and Its Genetic Diversity by ISSR and SRAP Profiling Analyses.

    Directory of Open Access Journals (Sweden)

    Na Liu

    Full Text Available Blumeria graminis f. sp. tritici, which causes wheat powdery mildew, is an obligate biotrophic pathogen that can easily genetically adapt to its host plant. Understanding the virulence structure of and genetic variations in this pathogen is essential for disease control and for breeding resistance to wheat powdery mildew. This study investigated 17 pathogenic populations in Sichuan, China and classified 109 isolates into two distinct groups based on pathogenicity analysis: high virulence (HV, 92 isolates and low virulence (LV, 17 isolates. Populations from Yibin (Southern region, Xichang (Western region, and Meishan (Middle region showed lower virulence frequencies than populations from other regions. Many of the previously known resistance genes did not confer resistance in this study. The resistance gene Pm21 displayed an immune response to pathogenic challenge with all populations in Sichuan, and Pm13, Pm5b, Pm2+6, and PmXBD maintained resistance. AMOVA revealed significantly higher levels of variation within populations and lower levels of variation among populations within regions. High levels of gene flow were detected among populations in the four regions. Closely related populations within each region were distinguished by cluster analyses using ISSR and SRAP alleles. Both ISSR and SRAP allele profiling analyses revealed high levels of genetic diversity among pathogenic populations in Sichuan. Although ISSR and SRAP profiling analysis showed similar resolutions, the SRAP alleles appeared to be more informative. We did not detect any significant association between these alleles and the virulence or pathogenicity of the pathogen. Our results suggest that ISSR and SRAP alleles are more efficient for the characterization of small or closely related populations versus distantly related populations.

  6. 南方红豆杉ISSR -PCR反应体系的建立与优化%An orthogonal design to optimize ISSR-PCR reaction system for Taxus wallichiana var.chinensis

    Institute of Scientific and Technical Information of China (English)

    廖盼华; 汪庆; 李亚; 姚淦; 杨如同; 王淑安

    2012-01-01

    Several important parameters influencing ISSR-PCR amplification on DNA from Taxus wallichiana var. Chinensis were studied with an orthogonal design by four factors and four levels to establish the optimum ISSR reaction system. The optimal ISSR-PCR reaction system was established as a total volume of 20μL consisting of 60ng template DNA, 10 x PCR buffer 2. 0μX, 25 mmol/L MgCl2 l.0μX, 2.5 mmol/L dNTPs l.0μL, 10μmol/L primer 1.0 uX and Taq DNA polymerase 0.2U. The suitable PCR procedure is one cycle denaturing at 94℃ for 4min,then, 41 cycles which involves denaturing at 941 for 30s,annealing at 54.5℃ for 45s and extending at 72℃ for 80s, one cycle extending at 72℃ for 5 min.and kept at4℃o%以南方红豆杉DNA为模板,采用正交试验设计对影响南方红豆杉ISSR -PCR扩增的参数进行了优化.结果表明较佳的南方红豆杉ISSR-PCR的反应体系(20 μL)为模板DNA 60 ng,10×PCR buffer 2.0μL,25 mmol/L的MgCl2 1.0 μL,2.5 mmol/L的dNTPs 1.0 μL,10μmol/L的引物1.0 μL和反应酶0.2U.适宜的扩增条件为94℃预变性4 min,41个PCR循环(94℃变性30 s,54.5℃退火45 s),72℃延伸80 s,72℃延伸5 min,4℃保存.

  7. Optimization of ISSR-PCR Reaction System on Ranunculus nephelogenes var. nephelogenes and Primer Selection%云生毛茛 ISSR-PCR 体系优化与引物筛选

    Institute of Scientific and Technical Information of China (English)

    石琳; 胡延萍; 王建科; 王钧; 许小宁; 李毅; 王莉

    2016-01-01

    This work is aimed to establish a stable ISSR-PCR system for Ranunculus nephelogenes var. nephelogenes. A L16(45) orthogonal design was used to screen 5 main parameters(Mg2+,dNTP,Taq DNA polymerase,primer,and template DNA);then the process of ISSR-PCR reaction was optimized,and consequently the optimal reaction system and amplifying procedure for R. nephelogenes var. nephelogenes was established ;further,the optimized reaction system and amplifying procedure was verified. Results showed that the optimal concentrations in 20 μL reaction mixture were template DNA 30 ng,Mg2+ 1.95 mmol/L,Taq DNA polymerase 0.04 U/μL,dNTP 0.150 mmol/L, and primer 0.5 μmol/L. The optimal reaction procedure was :pre-denaturalization for 5 mins at 94℃,denaturalization for 20 s at 94℃, renaturation for 1 min at 49.6-60.6℃,followed by 38 cycles of extension for 100 s at 72℃,and final extension for 6 min at 72℃. Under the above optimized reaction system and procedure,16 amplified primers were screened from 100 ISSR primers,and the optimal annealing temperature for each primer was determined. Verification in R. nephelogenes var. nephelogenes of different populations with the optimized ISSR system confirmed that bands amplified were clear and steady,thus the established ISSR-PCR could favor further studies on the genetic diversity of R. nephelogenes var. nephelogenes.%旨在建立稳定可靠的云生毛茛 ISSR-PCR 反应体系。采用正交试验设计方法,对影响云生毛茛 ISSR-PCR 扩增结果的 Mg2+、dNTP、Taq DNA 聚合酶、引物、模板 DNA 五个因素进行优化筛选,对反应程序进行优化,建立适用于云生毛茛的最佳反应体系和扩增程序,并对反应体系和扩增程序进行验证;在此基础上筛选多态性好的 ISSR 引物,采用梯度法筛选各个引物的最适退火温度。结果表明,云生毛茛20μL ISSR-PCR 的最佳反应体系为:模板 DNA 30 ng,Mg2+1.95 mmol/L,Taq DNA 聚合酶0.04 U/μL,dNTP 0

  8. Genetic Diversity of Pinus nigra Arn. Populations in Southern Spain and Northern Morocco Revealed By Inter-Simple Sequence Repeat Profiles

    Directory of Open Access Journals (Sweden)

    Oussama Ahrazem

    2012-05-01

    Full Text Available Eight Pinus nigra Arn. populations from Southern Spain and Northern Morocco were examined using inter-simple sequence repeat markers to characterize the genetic variability amongst populations. Pair-wise population genetic distance ranged from 0.031 to 0.283, with a mean of 0.150 between populations. The highest inter-population average distance was between PaCU from Cuenca and YeCA from Cazorla, while the lowest distance was between TaMO from Morocco and MA Sierra Mágina populations. Analysis of molecular variance (AMOVA and Nei’s genetic diversity analyses revealed higher genetic variation within the same population than among different populations. Genetic differentiation (Gst was 0.233. Cuenca showed the highest Nei’s genetic diversity followed by the Moroccan region, Sierra Mágina, and Cazorla region. However, clustering of populations was not in accordance with their geographical locations. Principal component analysis showed the presence of two major groups—Group 1 contained all populations from Cuenca while Group 2 contained populations from Cazorla, Sierra Mágina and Morocco—while Bayesian analysis revealed the presence of three clusters. The low genetic diversity observed in PaCU and YeCA is probably a consequence of inappropriate management since no estimation of genetic variability was performed before the silvicultural treatments. Data indicates that the inter-simple sequence repeat (ISSR method is sufficiently informative and powerful to assess genetic variability among populations of P. nigra.

  9. Genetic diversity of Poa pratensis L. depending on geographical origin and compared with genetic markers

    Directory of Open Access Journals (Sweden)

    Magdalena Szenejko

    2016-09-01

    Full Text Available Background Poa pratensis is one of the most common species of meadow grass in Europe. Most cultivars of the species found in Poland were originally derived from its ecotypes. We compared the effectiveness of the RAPD and ISSR methods in assessing the genetic diversity of the selected populations of P. pratensis. We examined whether these methods could be useful for detecting a possible link between the geographical origin of a given population and its assessed genetic variation. Methods The molecular markers RAPD and ISSR were used and their efficiency compared using, inter alia, statistical multivariate methods (UPGMA and PCA. Results The low value of Dice’s coefficient (0.369 along with the significantly high percentage of polymorphic products indicates a substantial degree of genetic diversity among the studied populations. Our results found a correlation between the geographical origin of the studied populations and their genetic variations. For ISSR, which proved to be the more effective method in that respect, we selected primers with the greatest differentiating powers correlating to geographical origin. Discussion The populations evaluated in this study were characterized by a high genetic diversity. This seems to confirm the hypothesis that ecotypes of P. pratensis originating from different regions of Central Europe with different terrain structures and habitat conditions can be a source of great genetic variability.

  10. High efficiency regeneration and genetic stability analysis of somatic ...

    African Journals Online (AJOL)

    Administrator

    2011-09-07

    Sep 7, 2011 ... polymorphic DNA (RAPD), inter-simple sequence repeat (ISSR) molecular markers and flow cytometry. ... random amplified polymorphic DNA (RAPD), regeneration. .... Detailed information on the primers used in the RAPD and ISSR analysis of ..... monomorphic bands comparing all the regenerated plants.

  11. Genetic Variation Analyses of Apple Triploid Hybrid Progenies from‘Hanfu’ב4xGala’by ISSR and AFLP%‘寒富’ב四倍体嘎拉’苹果的三倍性杂交后代ISSR和AFLP分析

    Institute of Scientific and Technical Information of China (English)

    王玉霞; 李林光; 何平; 张丽杰; 董文轩

    2013-01-01

    Genetic variation of 7 triploid hybrid progenies of‘Hanfu’ב4xGala’ were investigated based on ISSR and AFLP molecular markers in this paper. The results showed that the percentage of polymorphic bands (PPB) of ISSR was 60.00%, observed number of alleles (Na) was 1.600, effective number of alleles (Ne) was 1.420, Nei's gene diversity (H) was 0.236 and Shannon's Information index (I) was 0.346. And these were lower than that of AFLP, which were 66.54%, 1.665, 1.421, 0.240 and 0.357. The UPMGA cluster showed that most of 7 triploid hybrid progenies were tend to the female parent‘Hanfu’ based on ISSR and AFLP. Compared with ISSR, AFLP is more useful to analyze the genetic diversity of apple triploid hybrid progenies, and these triploid hybrid progenies were maternal inheritance. And these results can provide a basis for parent selecting in triploid apple breeding.%  利用ISSR及AFLP分子标记技术研究了‘寒富’ב四倍体嘎拉’苹果的7份三倍性后代及其亲本的遗传变异。结果表明ISSR扩增得到的遗传多样性指数中扩增多态性比率(PPB)为60.00%,观察等位基因数(Na)为1.600,有效等位基因数(Ne)为1.420,Nei's基因多样性指数(H)为0.236,Shannon信息指数(I)为0.346;均小于AFLP扩增的遗传多样性指数66.54%、1.665、1.421、0.240和0.357。根据ISSR和AFLP得到的UPGMA聚类图显示这7份三倍性杂交后代倾向于母本‘寒富’。在三倍性苹果遗传多样性检测上AFLP优于ISSR,且这7份三倍性新种质在遗传上倾向于母本遗传。为三倍性苹果育种的亲本选择提供依据。

  12. Deployment Repeatability

    Science.gov (United States)

    2016-04-01

    controlled to great precision, but in a Cubesat , there may be no attitude determination at all. Such a Cubesat might treat sun angle and tumbling rates as...could be sensitive to small differences in motor controller timing. In these cases, the analyst might choose to model the entire deployment path, with...knowledge of the material damage model or motor controller timing precision. On the other hand, if many repeated and environmentally representative

  13. ISSR Analysis of Genetic Relationships between Nine Varieties of Rose%9个油用玫瑰品种遗传关系的ISSR分析

    Institute of Scientific and Technical Information of China (English)

    高蕾; 姚雷

    2010-01-01

    研究利用ISSR(inter-simple sequence repeat)分子标记技术对栽培在我国的9个主要油用玫瑰品种的遗传关系进行了分析,从分子水平上阐明其遗传相似性,为建立油用玫瑰品种资源评价体系提供依据.结果表明,8个引物共扩增出135条带,其中123条为多态性带,多态性比率为91.11%,平均遗传相似系数为0.66,两两间的相似系数分布在0.398到1之间.聚类分析结果显示9个品种归为2组,其中第1组中,玫瑰2(Rosa damascena"HuZhou"),玫瑰3(Rosa damascena"PingYin")归为一亚组,玫瑰1(Rosa damascena"HuZhou")归为一亚组.第2组中玫瑰4(Rosa rugosa var.typica Reg)、玫瑰8(Rosa rugosa"FengHua")归为一亚组,玫瑰5(Rosa rugosa"LengXiang")、玫瑰6(Rosa rugosa"ZiZhi")、玫瑰9(Rosa rigosa"ZiZhi")归为一亚组,玫瑰7(Rosa serta×Rosa rugosa)单为一亚组.

  14. STR MARKERS. GENOTYPING APPLICATIONS

    Directory of Open Access Journals (Sweden)

    I. O. Sirbu

    2001-01-01

    Full Text Available STR (short tandem repeats loci consist of short, repetitive sequence elements of 2-8 bp in length. These abundant repeats are well distributed throughout the human genome and are rich source of highly polymorphic markers. There are literally hundreds of STR systems which have been mapped throughout the human genome. Several dozen have been investigated for application to human identity testing. These STR loci are found on almost every chromosome in the genome. They may be amplified using a variety of PCR primers. Tetranucleotide repeats have been most popular among forensic scientists due to their fidelity in PCR amplification although some tri- and pentanucleotide repeats are also in use. In this paper we intend (far from being exhaustive to present a synthesis of the characteristics of these genetic markers and their applications in genotyping, giving as an example the use of the STRs in a paternity testing case.

  15. A comparative analysis of genetic diversity in bitter gourd (Momordica charantia L.) genotypes using RAPD and ISSR markers

    Science.gov (United States)

    Bitter gourd (Momordica charantia L.) or bitter melon is a cucurbit of major economic importance where it is widely cultivated (India, China, Africa, and South America). The morphology (i.e., growth habit, maturity, and fruit shape, size, colour and surface texture) of Indian M. charantia germplasm...

  16. The PCR based technique RAPD is popular not only because it ...

    African Journals Online (AJOL)

    renu.bhatnagar

    2015-04-01

    Apr 1, 2015 ... RAPD marker analysis, 147 out of 222 bands (63.85%) were polymorphic and the ... Inter-simple sequence repeat (ISSR) primers are ..... Pearson correlation coefficient (r) estimates between genetic distances (GD) obtained.

  17. Molecular markers: a potential resource for ginger genetic diversity studies.

    Science.gov (United States)

    Ismail, Nor Asiah; Rafii, M Y; Mahmud, T M M; Hanafi, M M; Miah, Gous

    2016-12-01

    Ginger is an economically important and valuable plant around the world. Ginger is used as a food, spice, condiment, medicine and ornament. There is available information on biochemical aspects of ginger, but few studies have been reported on its molecular aspects. The main objective of this review is to accumulate the available molecular marker information and its application in diverse ginger studies. This review article was prepared by combing material from published articles and our own research. Molecular markers allow the identification and characterization of plant genotypes through direct access to hereditary material. In crop species, molecular markers are applied in different aspects and are useful in breeding programs. In ginger, molecular markers are commonly used to identify genetic variation and classify the relatedness among varieties, accessions, and species. Consequently, it provides important input in determining resourceful management strategies for ginger improvement programs. Alternatively, a molecular marker could function as a harmonizing tool for documenting species. This review highlights the application of molecular markers (isozyme, RAPD, AFLP, SSR, ISSR and others such as RFLP, SCAR, NBS and SNP) in genetic diversity studies of ginger species. Some insights on the advantages of the markers are discussed. The detection of genetic variation among promising cultivars of ginger has significance for ginger improvement programs. This update of recent literature will help researchers and students select the appropriate molecular markers for ginger-related research.

  18. Origin and Evolution of Jute Analysed by SRAP and ISSR Methods%SRAP结合ISSR方法分析黄麻属的起源与演化

    Institute of Scientific and Technical Information of China (English)

    陶爱芬; 祁建民; 李木兰; 方平平; 林荔辉; 徐建堂

    2012-01-01

    [Objective] The objective of this study is to make clear the origin and evolution of Corchorus with two molecular methods (SRAP and ISSR). [Method] Ninety-six jute germplasms were analysed with sequence related amplified polymorphism (SRAP) combining with inter-simple sequence related (ISSR) method. The phylogenetic trees of Corchorus were constructed by MEGA and DPS software, and the divergence time of jute germplasm was calculated. [Result] The relative wild species located at the basic position of the dendrogram, and the divergence time of which was the longest, which indicated the relative wild species originated earliest and was the ancestors of cultivated jute. Africa was the center of origin of Corchorus, while China was the second original center of Corchorus. Among all the olitorius species, the divergence time of wild and cultivated species from Africa was the longest, so Africa was the center of origin for wild and cultivated olitorius species, while the areas which border on India, Burma and China was the second center of origin for cultivated olitorius species. South China, countries of South Asia and Southeast Asia which border on China were the origin center of wild capsularis species, and South region of China was the origin center of cultivated capsularis species. Taken together, the divergence time of the cultivated capsularis species was shorter than olitorius species, indicating this biotype originated later than olitorius species. [ Conclusion ] Africa has an important position in origin of Corchorus, which was the primary origin center of wild species, wild and cultivated olitorius species. South region of China was the origin center of cultivated capsularis species. The divergence time and phylogenetic trees of Corchorus were calculated more comprehensively by SRAP markers combining with ISSR molecular markers, and the scientific conclusion on origin and evolution of Corchorus was got with above methods.%[目的]以SRAP结合ISSR分子标记

  19. 无患子基因组DNA提取及ISSR-PCR反应体系的建立与优化%DNA Extraction and Optimization of ISSR-PCR Reaction System for Sapindus Mukorossi

    Institute of Scientific and Technical Information of China (English)

    姜翠翠; 卢新坤; 方智振; 郭有枝; 姜业先; 刘献勇; 叶新福

    2013-01-01

    应用改良CTAB法提取了无患子基因组DNA,该方法具有操作过程简单、耗时少、能有效降低无患子基因组DNA提取过程中酚类化合物、多糖等杂质的污染.正交试验研究了dNTP浓度、引物浓度、Taq DNA聚合酶这3个因素对ISSR-PCR反应的影响,建立并优化了适合于无患子ISSR-PCR的反应体系.优化的反应总体系20μl含1 ×PCR Buffer,0.4 mmol· L-1 dNTP,0.8μmol· L-1引物,0.75 U Taq DNA聚合酶,20 ng DNA模板.使用此ISSR扩增反应体系,获得了部分不同种质无患子DNA的清晰条带,验证了该体系的稳定性.优化的反应体系为后续无患子遗传多样性的研究奠定了基础.

  20. Micropropagation of Pithecellobium dulce (Roxb.) Benth-a multipurpose leguminous tree and assessment of genetic fidelity of micropropagated plants using molecular markers.

    Science.gov (United States)

    Goyal, Pooja; Kachhwaha, Sumita; Kothari, S L

    2012-04-01

    An efficient and reproducible protocol has been developed for in vitro propagation of Pithecellobium dulce (Roxb.) Benth (a multipurpose leguminous tree) from field grown nodal segments (axillary bud). Shoot bud induction occurred from nodal explants of 15-years-old tree on Murashige and Skoog (MS) basal medium supplemented with 4.4 μM 6-benzyladenine (BA) and multiplication was achieved on MS medium supplemented with 4.4 μM BA + 0.73 μM phenylacetic acid (PAA) i.e. up to 7 shoot buds in the period of 5-6 weeks. Addition of adenine sulphate (AdS) to this medium further enhanced the number of shoot buds up to 10. Proliferating shoot cultures were established by repeatedly subculturing primary culture on fresh medium (MS + 4.4 μM BA + 0.73 μM PAA) after every 25 days. In vitro rooting was achieved on MS medium supplemented with 2.46 μM Indole-3-butyric acid (IBA) + 41.63 μM activated charcoal (AC). The micropropagated shoots with well developed roots were acclimatized in green house in pots containing sand, soil and manure (1:1:1). Genetic stability of micropropagated clones was evaluated using Random amplified polymorphic DNA (RAPD) and Inter simple sequence repeat (ISSR) markers. The amplification products were monomorphic in micropropagated plants and similar to those of mother plant. No polymorphism was detected revealing the genetic uniformity of micropropagated plants. This is the first report of an efficient protocol for regeneration of P. dulce through organogenesis, which can be used for further genetic transformation and pharmaceutical purposes.

  1. Short Tandem Repeat DNA Internet Database

    Science.gov (United States)

    SRD 130 Short Tandem Repeat DNA Internet Database (Web, free access)   Short Tandem Repeat DNA Internet Database is intended to benefit research and application of short tandem repeat DNA markers for human identity testing. Facts and sequence information on each STR system, population data, commonly used multiplex STR systems, PCR primers and conditions, and a review of various technologies for analysis of STR alleles have been included.

  2. Evaluation of efficiency of three molecular marker techniques for identifying poisonous plants%三种分子标记技术对有毒植物鉴定效能的评价

    Institute of Scientific and Technical Information of China (English)

    龙鑫; 孙承业; 谢立璟; 王英伟; 刘冰

    2013-01-01

    Objective To evaluate the efficiency of the three molecular marker techniques for identification of plant genus in order to select the most suitable molecular technique to rapidly identify a poisonous plant at the scene of the poisoning accident.Methods Nineteen samples of leaves in 18 species of common poisonous plants of Ranunculaceae and Euphorbiaceaec were selected.Genomic DNA of these samples were extracted using improved hexadecyltrimethylammonium bromide (CTAB) method,species identification (genus level) were performed using random amplified polymorphic DNA (RAPD),intersimple sequences repeats (ISSR) and DNA barcoding,cluster analysis were performed using NYSTS,SPSS,PAUP,and MEGA software.Accuracy,reliability,timeliness,and operability were compared between the 3 molecular marker techniques.Results Accuracy:genus level of samples failed to be identified using RAPD ; and the accuracy rates in genus level of identification using ISSR and DNA barcoding were 68% and 100%,respectively.Reliability:the subjective influence rates and the repetitive rates in RAPD and ISSR were 47% and 26% as well as 47% and 45%,respectively; the success rates of amplification and sequencing of trnH-psbA fragment using DNA barcoding both were 100%.Timeliness:the time from extraction of DNA to accomplishment of identification in the 3 molecular marker techniques were 8.5,9.0,and 42.2 h,respectively.Operability:species identification using RAPD and ISSR could be done in a common laboratory; special sequencing equipment was needed when using DNA barcoding technique in the identification of poisonous plants.Conclusion The preliminary study results show that DNA barcoding is a molecular identification technique more suitable for rapid determination of the poisonous plant at the scene of the sudden poisoning accident.%目的 评价3种不同的分子标记技术对植物属级的鉴定能力以选出最适合在中毒现场快速鉴定有毒植物的分子技术. 方法

  3. ProtRepeatsDB: a database of amino acid repeats in genomes

    Directory of Open Access Journals (Sweden)

    Chauhan Virander S

    2006-07-01

    repeat markers, interspecies variations and polymorphism.

  4. Studies on the Selective Parasitism Mechanism of Mites to Drone and Drone-Pupa by ISSR%运用ISSR分析蜂螨对雄蜂及雄蜂蛹的选择性寄生机制

    Institute of Scientific and Technical Information of China (English)

    刘益波; 吴小波; 颜伟玉; 曾志将

    2010-01-01

    以被大蜂螨(Varroa destructor)寄生的意大利蜜蜂(Apis mellifera ligustica)为实验材料,分别采集有螨雄蜂、无螨雄蜂、有螨雄蜂蛹和无螨雄蜂蛹各40只,运用ISSR(inter-simple sequence repeat)标记对各样品进行DNA扩增,通过fine-dox3型凝胶成像系统分析各条带的片段大小,计算各特异性条带在有螨与无螨样品中的概率.结果表明:有些特异性条带在有螨与无螨样品中的概率差异极显著(P<0.01),由此推测,这些条带可能与蜂螨的选择性寄生有关.

  5. 运用ISSR分析蜂螨对不同亚家系的选择性寄生%Study on the Selective Parasitism of the Varroa Mite to the Different Subfamilies by ISSR

    Institute of Scientific and Technical Information of China (English)

    刘益波; 黄强; 颜伟玉; 谢宪兵; 曾志将

    2009-01-01

    以被大蜂螨(Varroa destructor)寄生的意大利蜜蜂(Apis mellifera Ligustica)实验材料,分别采集有螨工蜂、无螨工蜂、有螨工蜂蛹和无螨工蜂蛹各36只,运用ISSR(in-ter-simple sequence repeat)标记和NTSYS聚类软件计算各个样品间的亲缘关系指数,并分析大蜂螨在寄生时是否存在亚家系趋向性.结果表明:144个样品分别分布在9个亚家系中,蜂螨在各个亚家系中的分布差异极显著(P<0.01),说明蜂螨在选择性寄生过程中存在亚家系的趋向性.

  6. Marker development

    Energy Technology Data Exchange (ETDEWEB)

    Adams, M.R.

    1987-05-01

    This report is to discuss the marker development for radioactive waste disposal sites. The markers must be designed to last 10,000 years, and place no undue burdens on the future generations. Barriers cannot be constructed that preclude human intrusion. Design specifications for surface markers will be discussed, also marker pictograms will also be covered.

  7. Establishment and optimization of ISSR-PCR and SRAP-PCR reaction systems of Thymus L.%百里香属植物ISSR-PCR和SRAP-PCR 体系的确立及优化

    Institute of Scientific and Technical Information of China (English)

    权俊萍; 袁菊红; 穆红梅; 张明霞; 李乃伟; 夏冰

    2008-01-01

    通过对 PCR 扩增体系中的模板 DNA 用量,Mg2+、dNTPs和引物浓度的单因子实验,分别建立了适合百里香属(Thymus L.)植物总 DNA 的 ISSR-PCR 及 SRAP-PCR 优化反应体系.ISSR-PCR 优化反应体系为:反应总体积20μL,含模板DNA 30 ng、Mg2+ 3.75 mmol·L-1、dNTPs 0.20 mmol·L-1、引物0.3 mmol·L-1和Taq DNA 聚合酶1 U.SRAP-PCR 优化反应体系为:反应总体积20μL,含模板DNA 30 ng、Mg2+ 2.50 mmol·L-1、dNTPs 0.10mmol·L-1、引物0.4 mmol·L-1和 Taq DNA 聚合酶l U.运用优化的反应体系对百里香属植物地椒(T. quinque-costatus Celak.)、地椒的亚洲变种[T. quinquecostatus var.asiaticus(Kitagawa)C.Y.Wu et Y.C.Huang]和百里香(T. mongolicus Ronn.)15个居群的总DNA进行了ISSR和SRAP初步分析,获得了较好的扩增结果.

  8. Agrobiodiversity in Cucurbita spp. landraces collected in Rio de Janeiro assessed by molecular markers

    Directory of Open Access Journals (Sweden)

    Marilene Hilma dos Santos

    2012-01-01

    Full Text Available Diversity and genetic relationship in forty landraces of Cucurbita spp. collected at small farms in Rio de Janeiro, Brazil, were analyzed by RAPD and ISSR markers, using 20 and 15 primers, respectively. Both markers were efficient to cluster the accessions separating among species, but not so much to the detection of intra-specific variability, considering the event of different pairs of accessions comprising null genetic distances observed for both markers in C. moschata. Low values observed for genetic distance among the C. moschata landraces showed that most likely genetic losses is in progress in that region of cultivation due to anthropic and market pressure, which are stimulating the small farmers to abandon their local varieties in order to use commercial seeds, including hybrids, which is causing risk of genetic erosion.

  9. A prospective study evaluating the performance of first trimester combined screening for trisomy 21 using repeated sampling of the maternal serum markers PAPP-A and free β-hCG

    DEFF Research Database (Denmark)

    Ekelund, Charlotte Kvist; Wright, Dave; Ball, Susan

    2012-01-01

    To prospectively evaluate the performance of first-trimester combined screening for trisomy 21 using the biochemical markers pregnancy-associated plasma protein-A (PAPP-A) and free beta-human chorionic gonadotropin (free β-hCG) obtained before and at the time of the nuchal translucency (NT) scan....

  10. Marker chromosomes.

    Science.gov (United States)

    Rao, Kiran Prabhaker; Belogolovkin, Victoria

    2013-04-01

    Marker chromosomes are a morphologically heterogeneous group of structurally abnormal chromosomes that pose a significant challenge in prenatal diagnosis. Phenotypes associated with marker chromosomes are highly variable and range from normal to severely abnormal. Clinical outcomes are very difficult to predict when marker chromosomes are detected prenatally. In this review, we outline the classification, etiology, cytogenetic characterization, and clinical consequences of marker chromosomes, as well as practical approaches to prenatal diagnosis and genetic counseling.

  11. Optimization of DNA extraction for ISSR studies in Tectona grandis Lf

    African Journals Online (AJOL)

    GRACE

    2006-07-03

    Jul 3, 2006 ... INTRODUCTION. Molecular marker studies require large amount of quality ... and high-density genome mapping (Godwin et al., 1997). MATERIAL .... high metabolic turnover requiring de novo synthesis of transcripts (RNA).

  12. Utilização de marcadores ISSR na avaliação da divergência genética entre acessos de biribazeiro

    Directory of Open Access Journals (Sweden)

    Rodrigo Monte Lorenzoni

    2014-01-01

    Full Text Available O biribazeiro é uma planta frutífera nativa das matas Atlântica e Amazônica. Seus frutos têm grande aceitação popular para consumo in natura. Objetivou-se com este estudo a avaliação da diversidade genética de acessos de biribazeiro (Rollinia mucosa [Jacq.] Baill com a utilização de marcadores moleculares ISSR. Foram analisados 16 acessos com 20 primers ISSR, os quais produziram um total de 118 bandas, sendo 96 polimórficas e 22 monomórficas. Os valores de dissimilaridade genética, calculados de acordo com o complemento do índice de Jaccard, variaram de 0,0909 a 0,5147. O método UPGMA (Unweighted Pair Group Method Average agrupou os acessos em seis grupos. Os acessos 1 e 5 foram mais dissimilares e 11 e 12 os menos dissimilares. Os marcadores ISSR utilizados neste estudo demonstraram eficiência na detecção de polimorfismos moleculares, revelando variabilidade genética entre os 16 acessos. Diante dos resultados obtidos neste trabalho, é possível inferir que existe considerável variabilidade genética entre os acessos de biribazeiro, demonstrando a importância dos marcadores na análise de variabilidade de espécies pouco estudadas, como Rollinia mucosa [Jacq.]Baill.

  13. Repeat-until-success quantum repeaters

    Science.gov (United States)

    Bruschi, David Edward; Barlow, Thomas M.; Razavi, Mohsen; Beige, Almut

    2014-09-01

    We propose a repeat-until-success protocol to improve the performance of probabilistic quantum repeaters. Conventionally, these rely on passive static linear-optics elements and photodetectors to perform Bell-state measurements (BSMs) with a maximum success rate of 50%. This is a strong impediment for entanglement swapping between distant quantum memories. Every time a BSM fails, entanglement needs to be redistributed between the corresponding memories in the repeater link. The key ingredients of our scheme are repeatable BSMs. Under ideal conditions, these turn probabilistic quantum repeaters into deterministic ones. Under realistic conditions, our protocol too might fail. However, using additional threshold detectors now allows us to improve the entanglement generation rate by almost orders of magnitude, at a nominal distance of 1000 km, compared to schemes that rely on conventional BSMs. This improvement is sufficient to make the performance of our scheme comparable to the expected performance of some deterministic quantum repeaters.

  14. 甘蔗物理诱变 M2代株系的 ISSR 分析%ISSR Analysis of Physically-mutated Sugarcane Strains in M2 Generation

    Institute of Scientific and Technical Information of China (English)

    廖芩; 黄河星; 黄兴; 杨善; 莫俊杰; 周鸿凯

    2015-01-01

    应用 ISSR 标记技术研究了海大32及其物理诱变 M2代33个株系的遗传多样性。结果表明:海大32及其 M2代株系的相似水平在0.86~1.00之间;在0.94左右的水平上,可以将它们分为4个大类,其中海大32及海大10-1等25个株系为第一类,海大10-3、海大10-27等6个株系聚为第二类,海大10-33单独为第三类,海大10-20和海大10-21聚为第四类;海大10-20、海大10-21发生变异的程度最大。%The author applied ISSR technique to study the genetic diversity of sugarcane variety Haida 32 and its physically-mutated 33 strains in M2 generation.The results showed that the similarity between Haida 32 and its M2-generation strains varied from 0.86 to 1.00.At the level of 0.94 or so, they could be divided into 4 types, the first type included 25 strains (Haida 32, Haida 10-1, and so on), the second type included 6 strains (Haida 10-3, Haida 10-27, etc.), the third type only included Haida 10-33, and the fourth type included Haida 10-20 and Haida 10-21.After physical mutation, the greatest degree of variations occurred in Haida 10-20 and Haida 10-21.

  15. Establishment and Optimization of ISSR-PCR Reaction System in Precocious Pyrus pyrifolia%早熟砂梨ISSR-PCR反应体系的建立与优化

    Institute of Scientific and Technical Information of China (English)

    宋燕春; 李永裕; 陈建烟; 黄添毅; 龚理; 吴少华

    2013-01-01

      以我国南方主栽的早熟砂梨品种‘翠冠’Pyrus pyrifolia‘Cuiguan’为材料,对ISSR技术体系中的模板DNA浓度、Taq DNA聚合酶用量、引物浓度、dNTP浓度、Mg2+浓度、退火温度、PCR循环数等7个主要因素进行优化和筛选,建立了适合早熟砂梨的ISSR-PCR反应体系。最终反应体系为20μL体系中10×PCR buffer (不含Mg2+)2μL,模板 DNA浓度60 ng,TaqDNA聚合酶0.75 U,引物浓度1μmol/L,dNTP浓度90μmol/L, Mg2+浓度2.25 mmol/L。扩增程序为:预变性94℃5 m in,变性94℃45 s,退火45 s,72℃延伸1 min,共42个循环,然后72℃再延伸10 min,4℃保存,用1.5%琼脂糖凝胶电泳检测多态性。%  To obtain 7 factors (DNA template dosage, Taq DNA polymerase dosage, primer concentration, dNTP concentration, Mg2+concentration, annealing temperature and PCR cycles ) of ISSR-PCR Reaction System in Pyrus pyrifolia ‘Cuiguan’, a precocious cultivar of southern China was been used as template. An optimum reaction system was been established. The final total volume was 20 μL, contains 2 μL 10×PCR buffer (Mg2+ free), 60 ng DNA template, 0.5 U Taq DNA polymerase, 1 μmol/L of primer concentration, 90 μmol/L of dNTP concentration, 2.25 mmol/L of Mg2+ concentration. Amplication procedure was as follows: initial-denaturing of 5 min at 94 ℃, followed by 42 cycles of 45 s for denaturing at 94℃, at annealing temperature for 45 s and 1 min at 72 ℃, after that 10 min extending at 72 ℃, and then keep at 4 ℃. Use 1.5% of agarose gel electrophoresis to test the genetic polymorphism.

  16. Prospective study evaluating performance of first-trimester combined screening for trisomy 21 using repeat sampling of maternal serum markers PAPP-A and free β-hCG.

    Science.gov (United States)

    Ekelund, C; Wright, D; Ball, S; Kirkegaard, I; Nørgaard, P; Sørensen, S; Friis-Hansen, L; Jørgensen, F S; Tørring, N; Bech, B H; Petersen, O B; Tabor, A

    2012-09-01

    To prospectively evaluate the performance of first-trimester combined screening for trisomy 21 using the biochemical markers pregnancy-associated plasma protein-A (PAPP-A) and free beta-human chorionic gonadotropin (free β-hCG) obtained before and at the time of the nuchal translucency (NT) scan. Three fetal medicine departments in Denmark participated in the study. Screening for trisomy 21 was set up as a two-step approach with blood sampling performed before the NT scan (early sample) and again at the time of the NT scan (late sample). PAPP-A and free β-hCG were measured on both the early and late samples. Age-standardized detection and false-positive rates for different screening protocols were calculated. We collected two blood samples in 27 pregnancies affected by trisomy 21 and in 3891 control pregnancies. The early samples were taken between gestational ages 8 + 0 and 13 + 6 weeks, and the late samples between 11 + 3 and 14 + 6 weeks. The median interval between the samples was 17 (range, 1-40) days. We found a significantly better estimated screening performance when using early sampling vs late sampling (P trisomy 21, use of early sampling with measurement of PAPP-A and free β-hCG before the time of the NT scan can optimize screening performance. Using maternal serum markers obtained both before and at the time of the NT scan has the potential to further improve performance, but larger studies are needed to confirm this potential. Copyright © 2012 ISUOG. Published by John Wiley & Sons, Ltd.

  17. Establishment and Optimization of Black Pepper ISSR Reaction System%胡椒ISSR反应体系的建立与优化

    Institute of Scientific and Technical Information of China (English)

    姜艳; 刘进平

    2012-01-01

    The conditions of ISSR-PCR reaction were optimized using the method of single-factor experiment respectively.The optimal ISSR-PCR amplification was established as follows: in a 20 μl reaction mixtures containing 1.0 U Taq DNA polymerase,0.8 mM of each dNTP,0.2 mM primer,1.8 mM Mg2+,and 50 ng template DNA for 35 thermal cycles.Amplifications were performed in an Biometra TGradient Thermal Cycler using the following conditions: in 20 μl reaction volumes for each primer,initial denaturation at 95℃ for 5min;denaturation at 94℃ for 30 s;annealing at the optimum anneal temperature selected for 30 s;extension at 72℃ for 45 s;35 cycles;the last cycle followed by 7 min extension at 72℃.This reaction system can be successfully applied to the analysis of genetic diversity of Piper plants in Hainan Island.%旨在构建胡椒(Piper nigrum L.)基因组DNA的ISSR-PCR反应体系,以便为海南胡椒属植物的遗传多样性分析研究打下基础。利用单因素随机试验设计,对胡椒ISSR-PCR反应体系中各组分(TaqDNA聚合酶、dNTPs、模板DNA、引物和Mg2+)的浓度进行优化,同时筛选ISSR-PCR反应的循环数和最适退火温度。确定了最优的ISSR-PCR反应体系为:总体积20μL,其中Taq DNA聚合酶1.0 U,dNTPs 0.8 mmol/L,引物0.2 mmol/L,Mg2+1.8 mmol/L,模板DNA 50 ng。同时通过梯度PCR试验,确定引物最佳退火温度及最佳循环数。胡椒ISSR-PCR最佳反应程序为:95℃预变性5 min;94℃变性30 s,54.9℃(引物834)退火30 s,72℃延伸45 s,共计35个循环,循环结束后72℃延伸7 min,4℃保存。这一反应体系可成功地应用于海南胡椒属植物的遗传多样性分析。

  18. (SSR) markers

    African Journals Online (AJOL)

    Yomi

    2012-04-03

    Apr 3, 2012 ... seeded and black-seeded cultivars and breeding lines. The group B included 70 ... maize, rice and tomatoes (Reif et al., 2006; Vigouroux et al., 2005; Warburton et ..... development of molecular markers for marker-assisted breeding. .... Selection under domestication: evidence for a sweep in the rice Waxy ...

  19. 诺丽叶片DNA的提取及ISSR-PCR 反应体系的建立%Isolation of Genomic DNA in Morinda citrifolia Linn. and Establishment of Its ISSR-PCR Reaction System

    Institute of Scientific and Technical Information of China (English)

    吴田; 蓝增全; 李青红

    2010-01-01

    [目的]提取诺丽(Morinda citrifolia Linn.)叶片DNA,并建立ISSR-PCR反应体系.[方法]以3份采自海南、4份采自美国的诺丽种质的叶片为材料,对其DNA提取和ISSR分子标记方法进行了研究.[结果]采用改进的CTAB DNA微量提取法,可以得到高质量的诺丽叶片基因组DNA.用14条不同的ISSR引物对所提取的诺丽基因组DNA进行了ISSR分子标记分析,其中7条引物在诺丽DNA中可扩增出多态性产物.[结论]建立了诺丽叶片基因组DNA快速、高效的提取方法和ISSR标记体系,可为ISSR分析应用于诺丽遗传研究奠定良好的基础.

  20. A Simple Sequence Repeat (SSR Marker Comparison of a Large In- and Ex-situ Potato Landrace Cultivar Collection from Peru Reaffirms the Complementary Nature of both Conservation Strategies

    Directory of Open Access Journals (Sweden)

    Jos van der Maesen

    2013-07-01

    Full Text Available An enhanced understanding of the temporal dynamics of intraspecific diversity is anticipated to improve the adequacy of conservation priorities, methods and metrics. We report on the comparative genetic composition of ex- and in-situ landrace cultivar populations from a potato diversity hotspot in the Andes. A total of 989 landrace cultivars belonging to contemporary custodian-farmer in situ collections from central Peru were compared with 173 accessions from a spatially analogous, but temporally differential ex situ composite genotype reference (CGR set using 15 nuclear microsatellite markers. A total of 173 alleles were detected, with 129 alleles (74.6% being shared between both populations. Both populations contain exclusive allelic diversity with 32 and 12 unique alleles belonging to the ex- and in-situ population, respectively. The mean unbiased expected heterozygosity values of the ex- and in-situ population are very similar, 0.749 versus 0.727, with a slightly wider range and standard deviation encountered for the in situ population. Analysis of Molecular Variance shows that 98.8% of the total variation is found within both populations, while the fixation index (Fst = 0.01236 corroborates that the populations are not well differentiated. Surprisingly, only 41.0% of the ex situ population encounters a similar landrace cultivar in 23.4% of the in situ population at a non-stringent threshold similarity coefficient of 0.80. While the ex- and in-situ population under comparison show similarities and unique features at the allelic level, their landrace cultivar composition is surprisingly distinct. Results affirm that crop evolution is an ongoing phenomenon and that change in fixed geographies is occurring.

  1. Bone Markers

    Science.gov (United States)

    ... markers may be seen in conditions such as: Osteoporosis Paget disease Cancer that has spread to the bone (metastatic bone disease) Hyperparathyroidism Hyperthyroidism Osteomalacia in adults and rickets in children—lack of bone mineralization, ...

  2. 怀地黄ISSR扩增条件优化的研究%Optimization of ISSR-PCR amplification in Huai Rehmannia glutinosa

    Institute of Scientific and Technical Information of China (English)

    周延清; 景建洲; 李振勇; 浩健; 贾敬芬

    2004-01-01

    Huai Rehmannia glutinosa Libosch. genomic DNA,extracted from its fresh young leaves by CTAB method,was used as ISSR-PCR amplification template.The effects of main elements,i.e.temperature,Taq polymerase dosage,dNTP concentration,Mg2+ concentration,primer concentration and template DNA concentration on ISSR-PCR amplication were tested by single factor experiment,respectively,to determine its optimal levels and establish the following optimal reaction system for ISSR analysis in Huai Rehmannia glutinosa Libosch.:PCR reaction volume of 25 μL,1.5~1.0 U Taq DNA polymerase,3.0 mmol/L Mg2+,1 × Taq DNA polymerase buffer (10 mmol/L Tris-HCl,50 mmol/L KCl,0.1% Trion X-100,pH9.0 ),60 ng template DNA,0.4 μmmol/L primer,0.4 mmol/L each of dATP,dGTP,dCTP and dTTP.Proper annealing temperature was 53~55℃.This has laid the good foundation for ISSR analysis under way in Huai Rehmannia glutinosa Libosch..%用CTAB法提取怀地黄嫩叶DNA,进行简单重复间序列标记(ISSR)分析.通过单因子实验分别研究了退火温度、Taq酶单位、Mg2+浓度、dNTP浓度、引物浓度和模板DNA浓度对ISSR-PCR反应的影响,找出各自的合适条件,而且每一个合适条件确定以后都被作为后续研究的一个条件.通过各个因子的组合研究建立了适宜于怀地黄ISSR分析的扩增体系:25 μL PCR反应体积,1×Taq DNA酶缓冲液(10 mmol/L Tris-HCl,50 mmol/L KCl,0.1% Trion X-100,pH9.0 ),2.5 mmol/L MgCl2,1.5~1.0 U Taq酶,60 ng模板DNA,0.4 μmmol/L引物,各0.4 mmol/L的dATP、dGTP、dCTP和dTTP.合适的退火温度为53~55℃.为用ISSR技术分析鉴定怀地黄种质资源奠定了良好的基础.

  3. OPTIMIZACIÓN DE UN PROTOCOLO DEL AISLAMIENTO DEL ADN Y DE UN SISTEMA DE AMPLIFICACIÓN ISSR-PCR PARA Ceratozamia mexicana Brongn. (Zamiaceae.

    Directory of Open Access Journals (Sweden)

    Nadia Guadalupe Sánchez-Coello

    2012-01-01

    Full Text Available La mayoría de las cícadas contienen altas concentraciones de aceites esenciales, flavonoides, polifenoles y polisacáridos que interfieren en la extracción de ADN, causando productos de amplificación errados o inhibiendo la PCR. La optimización del aislamiento del ADN y el empleo de iniciadores de secuencias intergénicas repetidas simples (ISSRs se investigaron en Ceratozamia mexicana Brongn., una cícada mexicana en peligro de extinción. El ADN obtenido de tejido foliar fresco, con un amortiguador modificado de cetil trimetil amonio, nos permitió obtener un ADN de buena calidad, sin pigmentos coloridos o contaminantes. La modificación al protocolo de extracción de ADN, basado en CTAB, fue un prelavado por 1 h, del tejido foliar, con una solución de 0.7 M de NaCl, para facilitar la lisis celular. El ADN extraído exitosamente se amplificó por PCR, usando seis iniciadores arbitrarios ISSR. Se observaron productos de amplificación reproducibles en todas las reacciones de PCR. Nuestros resultados muestran que la implementación mejora significativamente la calidad del ADN obtenido, usando una concentración baja de iniciadores (25 pM. Se detectaron 23 bandas fuertes, nueve de las cuales fueron polimórficas. Los resultados indican que el protocolo de optimización del aislamiento del ADN y en el sistema de PCR es viable para futuros trabajos en esta especie. Este trabajo es el primer protocolo de extracción de ADN y de ISSR reportado para esta especie ornamental en peligro de extinción.

  4. OPTIMIZACIÓN DE UN PROTOCOLO DEL AISLAMIENTO DEL ADN Y DE UN SISTEMA DE AMPLIFICACIÓN ISSR-PCR PARA Ceratozamia mexicana Brongn. (Zamiaceae).

    OpenAIRE

    Nadia Guadalupe Sánchez-Coello; Mauricio Luna-Rodríguez; Mario Vázquez-Torres; Lázaro Rafael Sánchez-Velásquez; Nancy Santana-Buzzy; Pablo Octavio-Aguilar; Lourdes Georgina Iglesias-Andreu

    2012-01-01

    La mayoría de las cícadas contienen altas concentraciones de aceites esenciales, flavonoides, polifenoles y polisacáridos que interfieren en la extracción de ADN, causando productos de amplificación errados o inhibiendo la PCR. La optimización del aislamiento del ADN y el empleo de iniciadores de secuencias intergénicas repetidas simples (ISSRs) se investigaron en Ceratozamia mexicana Brongn., una cícada mexicana en peligro de extinción. El ADN obtenido de tejido foliar fresco, con un amortig...

  5. 利用ISSR技术分析禾谷镰孢菌群体遗传多样性的研究%Study on Genetic Diversity of Fusarium graminearum Populations Causing Maize Stalk Rot by ISSR Analysis

    Institute of Scientific and Technical Information of China (English)

    何婧; 郭庆元; 王晓鸣; 宋利宁; 张维娜; 武小菲

    2011-01-01

    采用ISSR标记对采自我国11个省的玉米茎腐病相关禾谷镰孢的遗传多样性进行分析.利用筛选出的14个ISSR引物对供试的115株禾谷镰孢菌株进行扩增,共获得63条扩增清晰、重复性高的条带.其中多态性条带为60条,占95.2%.扩增条带分子量为150~2 000 bp,平均每个引物扩增出4.5条带.遗传多样性分析表明,在地理种群水平上,基因多样性指数在0~0.291 9之间,平均为0.159 1;Shannon‘s多样性指数在0~0.425 2之间,平均为0.236 0,表明不同地理种群间存在一定的遗传变异.多样性指数、等位基因数的增大与各种群内样本数量增加有关.遗传相似性分析证明,山东省种群与河南省种群间的遗传相似性最高,内蒙古种群与河北省种群间遗传相似性最低.在相似性系数为0.682时.可将115个菌株区分为2个聚类组,各组下又可分为3个亚组,分组结果与菌株的地理来源有一定相关性,表明禾谷镰孢的遗传分化与生态地理有关.%The genetic diversity of F. graminearum population collected from eleven provinces was determined using technique of inter-simple sequence repeat (ISSR). A total of 63 reproducible ISSR fragments were scored among 115 individuals, of which 60 (95.2%) were polymorphic. The size of the amplified fragments ranged from 150 bp to 2 000 bp and average number of bands per primer was 4.5. At geographic population level, Nei's gene diversity and Shannon's information index were 0.159 1 and 0.236 0, respectively, that meant there was genetic variation between geographic population of F. graminearum. By analysis of genetic similarity coefficient the F.graminearum populations of Shandong and Henan were closest and populations of Inner Mongolia and Hebei were the farthest. Based on genetic distances all isolates were clustered into two groups at the similarity of 0.682 and three subgroups in each group. The grouping results indicated that genetic variation of F

  6. 高加索三叶草、白三叶及其杂种F1代的ISSR分析%ISSR Analysis of Trifolium ambiguum Bieb., T.repens L.and Their Hybrid F1

    Institute of Scientific and Technical Information of China (English)

    黄帆; 王明玖; 何丽君; 陈丽丽; 魏春秋

    2012-01-01

    Trifolium ambiguum Bieb. (Caucasian clover, paternal) is an important species for the traits such as resistance to drought and cold; T. repens L. (white clover, maternal) is a species introduced from Greater Khingan for the traits of strong nitrogen fixation ability and growing ability. Hybridization was made between Caucasian clover and white clover and the embryos were successfully raised using the tissue culture technique, then the regeneration system of F1 was established. In order to define the genetic difference degree between hybrids F1 and their parents, ISSR molecular makers were used to analyze the genetic relationship. The results showed that 170 bands of ISSR markers were amplified with 16 primers, and the length of DNA fragment were ranged from 100 to 2000bp; 138 bands were polymorphic, and the percentage of polymorphism bands reached 81. 18%. The genetic distance (GD) of 4 tested plant materials ranged from 0. 0756 to 0. 6798, with an average of 0. 5313. The genetic distance between Caucasian clover and white clover was larger; the hybrid F1 was closer to Caucasian clover than to white clover, and there was a slight variation between F1 hybrids and regeneration of F1. Authenticities of hybrid F1 were identified and genetic differences between hybrids and parents were analyzed from the level of molecular.%利用耐寒、耐旱的六倍体高加索三叶草与强固氮能力、生长能力的大兴安岭四倍体白三叶进行远缘杂交,通过胚拯救技术得到杂种F1,同时通过组培快繁技术建立了F1的再生体系.为了明确杂种F1与亲本在DNA水平上的差异,建立适用于F1的分子标记反应体系,利用ISSR技术对其进行了鉴定分析.结果表明,16个ISSR引物共扩增出170条条带,长度介于100~2000bp之间,多态性条带138条,多态性条带百分率为81.18%;4个供试材料间的遗传距离变幅为0.0756~0.6798,平均为0.5313;父本白三叶与母本高加索三叶草

  7. Quantum repeated games revisited

    CERN Document Server

    Frackiewicz, Piotr

    2011-01-01

    We present a scheme for playing quantum repeated 2x2 games based on the Marinatto and Weber's approach to quantum games. As a potential application, we study twice repeated Prisoner's Dilemma game. We show that results not available in classical game can be obtained when the game is played in the quantum way. Before we present our idea, we comment on the previous scheme of playing quantum repeated games.

  8. Marker Recycling in Candida albicans through CRISPR-Cas9-Induced Marker Excision

    Science.gov (United States)

    2017-01-01

    ABSTRACT We describe here a new approach to marker recycling, a controlled sequence of steps in which a genetic marker is selected and then lost. Marker recycling is important for genetic manipulation, because it allows a single selection marker to be used repeatedly. Our approach relies upon the ability of the CRISPR-Cas9 system to make a targeted double-strand break in DNA and the expectation that a double-strand break within a selection marker may promote recombination between directly repeated sequences that flank the marker. We call the approach CRISPR-Cas9-induced marker excision (CRIME). We tested the utility of this approach with the fungal pathogen Candida albicans, which is typically diploid. We used two selection markers, modified to include flanking direct repeats. In a proof-of-principle study, we created successive homozygous deletions in three genes through use of the two markers and had one of the markers available in the final strain for further selection and recycling. This strategy will accelerate the creation of multiple-mutant strains in C. albicans. CRISPR-Cas9 systems have been applied to many organisms, so the genetic design principles described here may be broadly applicable. IMPORTANCE It is critical to be able to alter genes in order to elucidate their functions. These alterations often rely upon markers that allow selection for a rare cell in a population that has incorporated a piece of DNA. The number of alterations that can be accomplished is thus limited by the number of selection markers that are available. This limitation is circumvented by marker recycling strategies, in which a marker is eliminated after its initial use. Then, the marker can be used again. In this report, we describe a new marker recycling strategy that is enabled by recently developed CRISPR-Cas9 technology. PMID:28317025

  9. Optimization of DNA Extraction for RAPD and ISSR Analysis of Arbutus unedo L. Leaves

    Directory of Open Access Journals (Sweden)

    Paula Baptista

    2011-06-01

    Full Text Available Genetic analysis of plants relies on high yields of pure DNA. For the strawberry tree (Arbutus unedo this represents a great challenge since leaves can accumulate large amounts of polysaccharides, polyphenols and secondary metabolites, which co-purify with DNA. For this specie, standard protocols do not produce efficient yields of high-quality amplifiable DNA. Here, we present for the first time an improved leaf-tissue protocol, based on the standard cetyl trimethyl ammonium bromide protocol, which yields large amounts of high-quality amplifiable DNA. Key steps in the optimized protocol are the addition of antioxidant compounds—namely polyvinyl pyrrolidone (PVP, 1,4-dithiothreitol (DTT and 2-mercaptoethanol, in the extraction buffer; the increasing of CTAB (3%, w/v and sodium chloride (2M concentration; and an extraction with organic solvents (phenol and chloroform with the incubation of samples on ice. Increasing the temperature for cell lyses to 70 °C also improved both DNA quality and yield. The yield of DNA extracted was 200.0 ± 78.0 µg/µL and the purity, evaluated by the ratio A260/A280, was 1.80 ± 0.021, indicative of minimal levels of contaminating metabolites. The quality of the DNA isolated was confirmed by random amplification polymorphism DNA and by inter-simple sequence repeat amplification, proving that the DNA can be amplified via PCR.

  10. Genetic analysis and marker assisted identification of life phases of red alga Gracilaria corticata (J. Agardh).

    Science.gov (United States)

    Baghel, Ravi S; Kumari, Puja; Bijo, A J; Gupta, Vishal; Reddy, C R K; Jha, B

    2011-08-01

    The present study firstly reports the cytological and molecular marker assisted differentiation of isomorphic population of Gracilaria corticata (J. Agardh) with inter and intra-phasic genetic diversity analysis using ISSR markers. The genetic diversity of inbreeding population of G. corticata as determined in terms of percentage of polymorphic loci (PPL), average heterozygosity (He) and Shannon's Weaver index (I) were 59.80, 0.59 and 1.21, respectively. The inter-phasic pair-wise average polymorphism were found to be 31.6% between male and female, 24.0% in male and tetrasporophyte and 25.3% in female and tetrasporophyte. The intra-phasic average polymorphisms were calculated as a maximum of 5.5% between females, 4.2% between males and the lowest 2.4% between tetrasporophytes. The primer 10 generated a marker of 800 bp specific to male and 650 bp to female gametophyte, while the primer 17 generated a marker of 2,500 bp specific to tetrasporophyte. Both the UPGMA based dendrogram and PCA analysis clustered all the three life phases differentially as distinct identity. Cytological analysis by chromosome count revealed 24 chromosomes in both haploid male and female gametophytes (N) and 48 for diploid (2 N) tetrasporophyte further confirming their genetic distinctness. The life phase specific markers reported in this study could be of help in breeding programmes where differentiation of life phases at the early developmental stages is crucial.

  11. GENETIC DIVERSITY IN ACCESSIONS OF Passiflora cincinnata Mast. BASED ON MORPHOAGRONOMIC DESCRIPTORS AND MOLECULAR MARKERS

    Directory of Open Access Journals (Sweden)

    TIAGO VINÍCIUS BATISTA DO CARMO

    2017-01-01

    Full Text Available Passiflora cincinnata Mast. has become more popular in the market because the unusual flavor of its fruits and natural beauty of its flowers, and has great potential for breeding programs of Passiflora edulis f. flavicarpa, because its resistance to diseases and drought. The objective of this work was to evaluate seven wild passion fruit (P. cincinnata accessions, using morphological and agronomic descriptors and molecular markers type ISSR, to identify their morphoagronomic and genetic variabilities and potential for use in breeding programs. A randomized block experimental design was used with five replications and two plants per plot. Thirteen qualitative and twenty-one quantitative, vegetative and floral characteristics were used for morphoagronomic characterization. Twelve ISSR primers were evaluated for molecular characterization. Among the qualitative characteristics, only the color variations were significantly different between the accessions. According to the mean squares of the quantitative characteristics evaluated, obtained from analysis of variance, the means of accessions showed significant differences (p<0.01 for all characteristics. The IAL (internode average length was the morphological descriptor that most contributed to diversity, with 43.12%, followed by DH5 (stem diameter at 5 cm height and SW (sepal width. The average genetic similarity found was 68%. Despite the low genetic variability found among accessions, the primers UBC-887 and UBC-841 stood out with high percentage of polymorphism with 14 and 11 polymorphic fragments, respectively, and higher values of polymorphism information content (PIC, resolving power (RP and marker index (MI, denoting suitability for use in diversity studies of P. cincinnata. Low variability was found among accessions evaluated.

  12. Microsatellite markers spanning the apple (Malus x domestica Borkh.) genome

    NARCIS (Netherlands)

    Silfverberg-Dilworth, E.; Matasci, C.L.; Weg, van de W.E.; Kaauwen, van M.P.W.; Walser, M.; Kodde, L.P.; Soglio, V.; Gianfranceschi, L.; Durel, C.E.; Costa, F.; Yamamoto, T.; Koller, B.; Gessler, C.; Patocchi, A.

    2006-01-01

    A new set of 148 apple microsatellite markers has been developed and mapped on the apple reference linkage map Fiesta x Discovery. One-hundred and seventeen markers were developed from genomic libraries enriched with the repeats GA, GT, AAG, AAC and ATC; 31 were developed from EST sequences. Markers

  13. Microsatellite markers spanning the apple (Malus x domestica Borkh.) genome

    NARCIS (Netherlands)

    Silfverberg-Dilworth, E.; Matasci, C.L.; Weg, van de W.E.; Kaauwen, van M.P.W.; Walser, M.; Kodde, L.P.; Soglio, V.; Gianfranceschi, L.; Durel, C.E.; Costa, F.; Yamamoto, T.; Koller, B.; Gessler, C.; Patocchi, A.

    2006-01-01

    A new set of 148 apple microsatellite markers has been developed and mapped on the apple reference linkage map Fiesta x Discovery. One-hundred and seventeen markers were developed from genomic libraries enriched with the repeats GA, GT, AAG, AAC and ATC; 31 were developed from EST sequences. Markers

  14. Reconfigurable multiport EPON repeater

    Science.gov (United States)

    Oishi, Masayuki; Inohara, Ryo; Agata, Akira; Horiuchi, Yukio

    2009-11-01

    An extended reach EPON repeater is one of the solutions to effectively expand FTTH service areas. In this paper, we propose a reconfigurable multi-port EPON repeater for effective accommodation of multiple ODNs with a single OLT line card. The proposed repeater, which has multi-ports in both OLT and ODN sides, consists of TRs, BTRs with the CDR function and a reconfigurable electrical matrix switch, can accommodate multiple ODNs to a single OLT line card by controlling the connection of the matrix switch. Although conventional EPON repeaters require full OLT line cards to accommodate subscribers from the initial installation stage, the proposed repeater can dramatically reduce the number of required line cards especially when the number of subscribers is less than a half of the maximum registerable users per OLT. Numerical calculation results show that the extended reach EPON system with the proposed EPON repeater can save 17.5% of the initial installation cost compared with a conventional repeater, and can be less expensive than conventional systems up to the maximum subscribers especially when the percentage of ODNs in lightly-populated areas is higher.

  15. Revisiting the TALE repeat.

    Science.gov (United States)

    Deng, Dong; Yan, Chuangye; Wu, Jianping; Pan, Xiaojing; Yan, Nieng

    2014-04-01

    Transcription activator-like (TAL) effectors specifically bind to double stranded (ds) DNA through a central domain of tandem repeats. Each TAL effector (TALE) repeat comprises 33-35 amino acids and recognizes one specific DNA base through a highly variable residue at a fixed position in the repeat. Structural studies have revealed the molecular basis of DNA recognition by TALE repeats. Examination of the overall structure reveals that the basic building block of TALE protein, namely a helical hairpin, is one-helix shifted from the previously defined TALE motif. Here we wish to suggest a structure-based re-demarcation of the TALE repeat which starts with the residues that bind to the DNA backbone phosphate and concludes with the base-recognition hyper-variable residue. This new numbering system is consistent with the α-solenoid superfamily to which TALE belongs, and reflects the structural integrity of TAL effectors. In addition, it confers integral number of TALE repeats that matches the number of bound DNA bases. We then present fifteen crystal structures of engineered dHax3 variants in complex with target DNA molecules, which elucidate the structural basis for the recognition of bases adenine (A) and guanine (G) by reported or uncharacterized TALE codes. Finally, we analyzed the sequence-structure correlation of the amino acid residues within a TALE repeat. The structural analyses reported here may advance the mechanistic understanding of TALE proteins and facilitate the design of TALEN with improved affinity and specificity.

  16. Construction of a genetic linkage map of black gram, Vigna mungo (L.) Hepper, based on molecular markers and comparative studies.

    Science.gov (United States)

    Gupta, S K; Souframanien, J; Gopalakrishna, T

    2008-08-01

    A genetic linkage map of black gram, Vigna mungo (L.) Hepper, was constructed with 428 molecular markers using an F9 recombinant inbred population of 104 individuals. The population was derived from an inter-subspecific cross between a black gram cultivar, TU94-2, and a wild genotype, V. mungo var. silvestris. The linkage analysis at a LOD score of 5.0 distributed all 428 markers (254 AFLP, 47 SSR, 86 RAPD, and 41 ISSR) into 11 linkage groups. The map spanned a total distance of 865.1 cM with an average marker density of 2 cM. The largest linkage group spanned 115 cM and the smallest linkage group was of 44.9 cM. The number of markers per linkage group ranged from 11 to 86 and the average distance between markers varied from 1.1 to 5.6 cM. Comparison of the map with other published azuki bean and black gram maps showed high colinearity of markers, with some inversions. The current map is the most saturated map for black gram to date and will provide a useful tool for identification of QTLs and for marker-assisted selection of agronomically important characters in black gram.

  17. Recursive quantum repeater networks

    CERN Document Server

    Van Meter, Rodney; Horsman, Clare

    2011-01-01

    Internet-scale quantum repeater networks will be heterogeneous in physical technology, repeater functionality, and management. The classical control necessary to use the network will therefore face similar issues as Internet data transmission. Many scalability and management problems that arose during the development of the Internet might have been solved in a more uniform fashion, improving flexibility and reducing redundant engineering effort. Quantum repeater network development is currently at the stage where we risk similar duplication when separate systems are combined. We propose a unifying framework that can be used with all existing repeater designs. We introduce the notion of a Quantum Recursive Network Architecture, developed from the emerging classical concept of 'recursive networks', extending recursive mechanisms from a focus on data forwarding to a more general distributed computing request framework. Recursion abstracts independent transit networks as single relay nodes, unifies software layer...

  18. Shoot organogenesis in leaf explants of Hydrangea macrophylla ‘Hyd1’ and assessing genetic stability of regenerants using ISSR markers

    NARCIS (Netherlands)

    Liu, F.; Huang, L.L.; Reinhoud, P.; Jongsma, M.A.; Wang, C.

    2011-01-01

    For the first time, an in vitro regeneration protocol of Hydrangea macrophylla 'Hyd1' was developed. Effects of different plant growth regulators (PGRs) on shoot regeneration were investigated jointly with selecting optimal basal media and cefotaxime concentrations. The highest frequency of shoot or

  19. Identification of Relationship of Soybean Using ISSR Marker%利用ISSR技术探讨大豆属植物的亲缘关系

    Institute of Scientific and Technical Information of China (English)

    王玉民; 姜昱; 康岭生; 赵桂兰

    2007-01-01

    对大豆属Glycine亚属19份多年生野生大豆、Soja亚属2份野生大豆和2份栽培大豆进行了ISSR分析.结果表明:1.在Soja亚属中野生大豆和栽培大豆的指纹图谱很相近,相似性系数高达0.961,但在野生大豆和栽培大豆之间存在特异谱带.2.Glycine亚属二倍体材料中,染色体组型为A类的G.argyrea、G.canescens 和G.clandestina 很好地聚为一类;G.curvata和G.cyrtoloba 的染色体组型同为C类,其指纹图谱的相似程度也较高;B类染色体组型G.latifolia和G.tabacina(2n=40)的亲缘关系亦较近,同为B类染色体组型的G.microphylla却与染色体组型为C组的G.curvata和G.cyrtoloba的指纹图谱比较接近.3、Glycine亚属多倍体材料中在G.tomentella和G.tabacina种内的多倍体材料的亲缘关系较近,尽管其地理来源不同;染色体组型为DD的G.tomentella(2n=40)和染色体组型为EE的G.tomentella(2n=38)与其它多倍体种的亲缘关系较远.4.Soja亚属和Glycine亚属的相似性系数很低,亲缘关系较远,很早就发生了分化.

  20. Analysis on ISSR Markers of Germplasm for Cuminum cyminum L.in Xinjiang%新疆孜然种质资源的ISSR标记分析

    Institute of Scientific and Technical Information of China (English)

    马艳明; 刘志勇; 王浩; 韩稳稳; 热依拉木·木合甫力; 白玉亭

    2008-01-01

    应用ISSR标记对24份孜然种质资源进行DNA多态性分析.从38条引物中筛选出12条多态性引物,共扩增出118条带,其中39条多态性带;引物等位位点数在6~16个,平均每个引物扩增出9.83条带,引物等位位点多态性信息含量(PIC)变幅为11.11%~62.50%,平均35.14%.24个品种间的Nel's遗传相似系数变化范围为0.33~0.90,平均为0.73.聚类结果表明各类群与地域来源联系不明显,说明孜然种质资源在新疆范围内有比较频繁的交流.

  1. Optimal of ISSR Marker in Dactylis glomerata L.%鸭茅ISSR反应体系的建立与优化(简报)

    Institute of Scientific and Technical Information of China (English)

    曾兵; 张新全; 兰英

    2007-01-01

    简单序列重复区间扩增多态性(Inter—simple sequence repeat,简称ISSR)是由zietkiewicz等于1994年创建的一种DNA标记技术。ISSR技术的原理和操作与SSR、RAPD非常相似,只是引物设计要求不同,但其产物多态性比RFLP、SSR、RAPD丰富,可提供更多的基因组信息,比RAPD技术更加稳定可靠,实验重复性好。

  2. 木菠萝ISSR反应体系的建立和优化%Establishment and Optimization of an ISSR Reaction System in Jackfruit

    Institute of Scientific and Technical Information of China (English)

    王耀辉; 叶春海; 李映志; 丰锋

    2008-01-01

    以木菠萝为材料,研究了模板DNA、Taq DNA聚合酶、引物、Mg2+ 及dNTPs浓度对ISSR-PCR扩增结果的影响,得出木菠萝ISSR的较佳反应体系为:在20μI反应体系中DNA模板1.6 ng·μI-1,Taq DNA聚合酶1.25 U·(20μl)-1,引物0.24 μmol·L-1,dNTPs 0.2 mmol-1 ,Mg2+ 2 mmol· L-1,10×PCR缓冲液2.0μl.

  3. The Pentapeptide Repeat Proteins

    Energy Technology Data Exchange (ETDEWEB)

    Vetting,M.; Hegde, S.; Fajardo, J.; Fiser, A.; Roderick, S.; Takiff, H.; Blanchard, J.

    2006-01-01

    The Pentapeptide Repeat Protein (PRP) family has over 500 members in the prokaryotic and eukaryotic kingdoms. These proteins are composed of, or contain domains composed of, tandemly repeated amino acid sequences with a consensus sequence of [S, T,A, V][D, N][L, F]-[S, T,R][G]. The biochemical function of the vast majority of PRP family members is unknown. The three-dimensional structure of the first member of the PRP family was determined for the fluoroquinolone resistance protein (MfpA) from Mycobacterium tuberculosis. The structure revealed that the pentapeptide repeats encode the folding of a novel right-handed quadrilateral {beta}-helix. MfpA binds to DNA gyrase and inhibits its activity. The rod-shaped, dimeric protein exhibits remarkable size, shape and electrostatic similarity to DNA.

  4. Repeats in transforming acidic coiled-coil (TACC) genes.

    Science.gov (United States)

    Trivedi, Seema

    2013-06-01

    Transforming acidic coiled-coil proteins (TACC1, 2, and 3) are essential proteins associated with the assembly of spindle microtubules and maintenance of bipolarity. Dysregulation of TACCs is associated with tumorigenesis, but studies of microsatellite instability in TACC genes have not been extensive. Microsatellite or simple sequence repeat instability is known to cause many types of cancer. The present in silico analysis of SSRs in human TACC gene sequences shows the presence of mono- to hexa-nucleotide repeats, with the highest densities found for mono- and di-nucleotide repeats. Density of repeats is higher in introns than in exons. Some of the repeats are present in regulatory regions and retained introns. Human TACC genes show conservation of many repeat classes. Microsatellites in TACC genes could be valuable markers for monitoring numerical chromosomal aberrations and or cancer.

  5. Survey of simple sequence repeats in woodland strawberry (Fragaria vesca).

    Science.gov (United States)

    Guan, L; Huang, J F; Feng, G Q; Wang, X W; Wang, Y; Chen, B Y; Qiao, Y S

    2013-07-30

    The use of simple sequence repeats (SSRs), or microsatellites, as genetic markers has become popular due to their abundance and variation in length among individuals. In this study, we investigated linkage groups (LGs) in the woodland strawberry (Fragaria vesca) and demonstrated variation in the abundances, densities, and relative densities of mononucleotide, dinucleotide, and trinucleotide repeats. Mononucleotide, dinucleotide, and trinucleotide repeats were more common than longer repeats in all LGs examined. Perfect SSRs were the predominant SSR type found and their abundance was extremely stable among LGs and chloroplasts. Abundances of mononucleotide, dinucleotide, and trinucleotide repeats were positively correlated with LG size, whereas those of tetranucleotide and hexanucleotide SSRs were not. Generally, in each LG, the abundance, relative abundance, relative density, and the proportion of each unique SSR all declined rapidly as the repeated unit increased. Furthermore, the lengths and frequencies of SSRs varied among different LGs.

  6. Quasimonomorphic Mononucleotide Repeats for High-Level Microsatellite Instability Analysis

    Directory of Open Access Journals (Sweden)

    Olivier Buhard

    2004-01-01

    Full Text Available Microsatellite instability (MSI analysis is becoming more and more important to detect sporadic primary tumors of the MSI phenotype as well as in helping to determine Hereditary Non-Polyposis Colorectal Cancer (HNPCC cases. After some years of conflicting data due to the absence of consensus markers for the MSI phenotype, a meeting held in Bethesda to clarify the situation proposed a set of 5 microsatellites (2 mononucleotide repeats and 3 dinucleotide repeats to determine MSI tumors. A second Bethesda consensus meeting was held at the end of 2002. It was discussed here that the 1998 microsatellite panel could underestimate high-level MSI tumors and overestimate low-level MSI tumors. Amongst the suggested changes was the exclusive use of mononucleotide repeats in place of dinucleotide repeats. We have already proposed a pentaplex MSI screening test comprising 5 quasimonomorphic mononucleotide repeats. This article compares the advantages of mono or dinucleotide repeats in determining microsatellite instability.

  7. Repeating the Past

    Science.gov (United States)

    Moore, John W.

    1998-05-01

    As part of the celebration of the Journal 's 75th year, we are scanning each Journal issue from 25, 50, and 74 years ago. Many of the ideas and practices described are so similar to present-day "innovations" that George Santayana's adage (1) "Those who cannot remember the past are condemned to repeat it" comes to mind. But perhaps "condemned" is too strong - sometimes it may be valuable to repeat something that was done long ago. One example comes from the earliest days of the Division of Chemical Education and of the Journal.

  8. Study on Total DNA Extraction of Alsophila costularis Baker and Its ISSR-PCR Analysis%中华桫椤DNA提取及ISSR分析

    Institute of Scientific and Technical Information of China (English)

    应站明; 黎桂芳; 王海舟

    2009-01-01

    [目的]研究中华桫椤总DNA的提取方法.[方法]采用SDS法和改良CTAB法提取中华桫椤总DNA,通过电泳检测方法进行比较.[结果]用改良CTAB法得到的DNA浓度和纯度都较高,基本上无蛋白质、多糖等杂质.用改良CTAB法得到的总DNA进行ISSR-PCR扩增,谱带清晰,获得较好的效果.[结论]为提取高质量的DNA样品奠定了基础,并为探讨中华桫椤自然居群的遗传多样性和种群遗传结构及进一步保护和利用中华桫椤提供了分子遗传学依据.%[Objective] The research aimed to study the total DNA extraction methods of A. costularis Baker.[Method] To investigate the optimum DNA extraction method, SDS and modified CTAB methods for isolating genomic DNA from A. costularis Baker leaf were studied. The isolated DNA was examined by ultraviolet absorption test and agarose gelelectrophoresis. [Result] The results showed that the modified CTAB method was suitable for DNA isolation from A. costularis Baker plants. The quality of DNA could meet ISSR requirement even extracted once by the modified CTAB method. [Conclusion] The study can provide the basis for extracting high quality DNA sample, and can offer the molecular genetics reference for exploring the genetic diversity and the population genetic structure of A. costularis Baker and deeply protecting and utilizing A. costularis Baker.

  9. All-optical repeater.

    Science.gov (United States)

    Silberberg, Y

    1986-06-01

    An all-optical device containing saturable gain, saturable loss, and unsaturable loss is shown to transform weak, distorted optical pulses into uniform standard-shape pulses. The proposed device performs thresholding, amplification, and pulse shaping as required from an optical repeater. It is shown that such a device could be realized by existing semiconductor technology.

  10. Bidirectional Manchester repeater

    Science.gov (United States)

    Ferguson, J.

    1980-01-01

    Bidirectional Manchester repeater is inserted at periodic intervals along single bidirectional twisted pair transmission line to detect, amplify, and transmit bidirectional Manchester 11 code signals. Requiring only 18 TTL 7400 series IC's, some line receivers and drivers, and handful of passive components, circuit is simple and relatively inexpensive to build.

  11. High-throughput development of genome-wide locus-specific informative SSR markers in wheat

    Science.gov (United States)

    Although simple sequence repeat (SSR) markers are not new, they are still useful and often used markers in molecular mapping and marker-assisted breeding, particularly in developing countries. However, locus-specific SSR markers could be more useful and informative in wheat breeding and genetic stud...

  12. Identification of ISSR Primers

    African Journals Online (AJOL)

    vegetables in West Africa (Abiose, 1999). Among the different ... among African edible cucurbits (Dje et al., 2006). However .... Monitoring and Assessing Urban Encroachment into ... Agricultural Land - A Remote Sensing and GIS Based Study.

  13. Genetic and Chemical Profiling of Gymnema sylvestre Accessions from Central India: Its Implication for Quality Control and Therapeutic Potential of Plant

    Science.gov (United States)

    Verma, Ashutosh Kumar; Dhawan, Sunita Singh; Singh, Seema; Bharati, Kumar Avinash; Jyotsana

    2016-01-01

    Background: Gymnema sylvestre, a vulnerable plant species, is mentioned in Indian Pharmacopeia as an antidiabetic drug Objective: Study of genetic and chemical diversity and its implications in accessions of G. sylvestre Materials and Methods: Fourteen accessions of G. sylvestre collected from Central India and assessment of their genetic and chemical diversity were carried out using ISSR (inter simple sequence repeat) and HPLC (high performance liquid chromatography) fingerprinting methods Results: Among the screened 40 ISSR primers, 15 were found polymorphic and collectively produced nine unique accession-specific bands. The maximum and minimum numbers of amplicones were noted for ISSR-15 and ISSR-11, respectively. The ISSR -11 and ISSR-13 revealed 100% polymorphism. HPLC chromatograms showed that accessions possess the secondary metabolites of mid-polarity with considerable variability. Unknown peaks with retention time 2.63, 3.41, 23.83, 24.50, and 44.67 were found universal type. Comparative hierarchical clustering analysis based on foresaid fingerprints indicates that both techniques have equal potential to discriminate accessions according to percentage gymnemic acid in their leaf tissue. Second approach was noted more efficiently for separation of accessions according to their agro-climatic/collection site Conclusion: Highly polymorphic ISSRs could be utilized as molecular probes for further selection of high gymnemic acid yielding accessions. Observed accession specific bands may be used as a descriptor for plant accessions protection and converted into sequence tagged sites markers. Identified five universal type peaks could be helpful in identification of G. sylvestre-based various herbal preparations. SUMMARY Nine accession specific unique bandsFive marker peaks for G. sylvestre.Suitability of genetic and chemical fingerprinting Abbreviations used: HPLC: High Performance Liquid Chromatography, ISSR: Inter Simple Sequence Repeats, CTAB: Cetyl

  14. Duct Leakage Repeatability Testing

    Energy Technology Data Exchange (ETDEWEB)

    Walker, Iain [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Sherman, Max [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

    2014-01-01

    Duct leakage often needs to be measured to demonstrate compliance with requirements or to determine energy or Indoor Air Quality (IAQ) impacts. Testing is often done using standards such as ASTM E1554 (ASTM 2013) or California Title 24 (California Energy Commission 2013 & 2013b), but there are several choices of methods available within the accepted standards. Determining which method to use or not use requires an evaluation of those methods in the context of the particular needs. Three factors that are important considerations are the cost of the measurement, the accuracy of the measurement and the repeatability of the measurement. The purpose of this report is to evaluate the repeatability of the three most significant measurement techniques using data from the literature and recently obtained field data. We will also briefly discuss the first two factors. The main question to be answered by this study is to determine if differences in the repeatability of these tests methods is sufficient to indicate that any of these methods is so poor that it should be excluded from consideration as an allowed procedure in codes and standards.

  15. Construction of marker-free transplastomic plants.

    Science.gov (United States)

    Lutz, Kerry A; Maliga, Pal

    2007-04-01

    Because of its prokaryotic-type gene expression machinery, maternal inheritance and the opportunity to express proteins at a high level, the plastid genome (plastome or ptDNA) is an increasingly popular target for engineering. The ptDNA is present as up to 10,000 copies per cell, making selection for marker genes essential to obtain plants with uniformly transformed ptDNA. However, the marker gene is no longer desirable when homoplastomic plants are obtained. Marker-free transplastomic plants can now be obtained with four recently developed protocols: homology-based excision via directly repeated sequences, excision by phage site-specific recombinanses, transient cointegration of the marker gene, and the cotransformation-segregation approach. Marker excision technology will benefit applications in agriculture and in molecular farming.

  16. Genetic variation in oriental tobacco (Nicotiana tabacum L. by agro-morphological traits and simple sequence repeat markers Variação genética em tabaco oriental (Nicotiana tabacum L. por marcadores agro-morfológicos e traços simples de repetição de sequência

    Directory of Open Access Journals (Sweden)

    Reza Darvishzadeh

    2013-06-01

    Full Text Available The objectives of this study were to assess genetic diversity and determine differences between several oriental tobacco genotypes by examining both agro-morphological traits and molecular markers. Simple lattice design with two replications was used to evaluate 100 oriental tobacco genotypes. Analysis of variance manifested that there is high level of genetic diversity in oriental-type tobaccos based on morphological traits including number of leaf, days to 50% flowering, leaf length, leaf width, leaf fresh weight, leaf dry weight, stem height and stem girth. Classification of genotypes using agro-morphological data by means of un-weighted pair-group method using arithmetic average (UPGMA algorithm based on squared standardized Euclidean distances resulted four distinguishable groups that pursuit own geographical distribution. In the molecular marker investigations, a total of 13 simple sequence repeats (SSR primer pairs were used to determine polymorphism of the test germplasm. Thirty five alleles were scored at 13 SSR loci. The average number of alleles per locus (na and the effective allele number (Ae were 2.69 and 2.34, respectively. By using SSR data, pair wise Jaccard's similarity coefficients were produced. Grouping of genotypes via Jaccard's similarity coefficients and using UPGMA clustering method lead to three groups that had not any accommodated with own origins. Results reveled that there is not completely agreement for classification based on agro-morphological and SSR loci in oriental-type tobaccos. Because of non influence of environmental effects on molecular marker, heterotic groups based on SSR markers could be closer to reality.Os objetivos deste estudo foram avaliar a diversidade genética e determinar as diferenças entre diversos genótipos de tabaco oriental através tratos morfológicos e marcadores moleculares. O delineamento utilizado foi o látice simples com duas repetições e foram avaliados 100 genótipos de

  17. Multiplexed microsatellite markers for seven Metarhizium species

    Science.gov (United States)

    Cross-species transferability of 41 previously published simple sequence repeat (SSR) markers was assessed for 11 species of the entomopathogenic fungus Metarhizium. A collection of 65 Metarhizium isolates including all 54 used in a recent phylogenetic revision of the genus were characterized. Betwe...

  18. Investigating Genetic Diversity of Foeniculum Vulgare Mill using Molecular Markers

    Directory of Open Access Journals (Sweden)

    Omid Jadidi

    2016-06-01

    Full Text Available Medicinal plants are considered valuable genetic resources in Iran. One of these medicinal as well as spice plants is Foeniculum Vulgare Mill from Umbellifetae family used in different industries such as food, medicine, and cosmetics. It seems that due to different climate conditions in Iran this plant represents a high and valuable genetic diversity; therefore, management of genetic resources protection and obtaining information about genetic diversity will help awareness of evolution processes as well as genetic erosion of this valuable plant. Genetic diversity in local masses of Foeniculum Vulgare Mill can be investigated using molecule markers such as AFLP, RAPD, ISSR, SRAP, RFLP, and so on. In investigation of over 30 ecotype of local Foeniculum Vulgare Mill, different markers have shown that mean polymorphic content (PIC is about 36% and mean genetic diversity is estimated about 40% in different samples. Data obtained from molecule software analyses help to categorize Foeniculum Vulgare Mill genotype in different groups based on climate and geographical conditions. Principle components analysis (PCOA has also confirmed the results of cluster analysis. Dendrogram obtained by cluster analysis based on similarity coefficient of simple matching (SM and UPGMA algorithm can also categorize population of Foeniculum Vulgare Mill in different groups. Results of molecular variance analysis (AMOVA have shown that most genetic variance between geographical groups can be seen in populations. In general, according to investigations, there is a significant genetic diversity regarding agronomic and molecular traits of Foeniculum Vulgare Mill masses in Iran and knowing this genetic diversity will help in breeding programs, complementary studies, categorization, and so on.

  19. Statistical analysis of repeated outcomes of different types

    NARCIS (Netherlands)

    Musoro, Z.J.

    2016-01-01

    This thesis focused on analyzing data with multiple outcome variables. The motivating data sets comprised longitudinal markers of patients’ disease state (e.g. B cells and CD4+ T cell) as well as information on the time to an event (e.g. death) or (multiple) recurrent event times (e.g. repeated

  20. Repeatability of Cryogenic Multilayer Insulation

    Science.gov (United States)

    Johnson, W. L.; Vanderlaan, M.; Wood, J. J.; Rhys, N. O.; Guo, W.; Van Sciver, S.; Chato, D. J.

    2017-01-01

    Due to the variety of requirements across aerospace platforms, and one off projects, the repeatability of cryogenic multilayer insulation has never been fully established. The objective of this test program is to provide a more basic understanding of the thermal performance repeatability of MLI systems that are applicable to large scale tanks. There are several different types of repeatability that can be accounted for: these include repeatability between multiple identical blankets, repeatability of installation of the same blanket, and repeatability of a test apparatus. The focus of the work in this report is on the first two types of repeatability. Statistically, repeatability can mean many different things. In simplest form, it refers to the range of performance that a population exhibits and the average of the population. However, as more and more identical components are made (i.e. the population of concern grows), the simple range morphs into a standard deviation from an average performance. Initial repeatability testing on MLI blankets has been completed at Florida State University. Repeatability of five GRC provided coupons with 25 layers was shown to be +/- 8.4 whereas repeatability of repeatedly installing a single coupon was shown to be +/- 8.0. A second group of 10 coupons have been fabricated by Yetispace and tested by Florida State University, through the first 4 tests, the repeatability has been shown to be +/- 16. Based on detailed statistical analysis, the data has been shown to be statistically significant.

  1. Establishment and Optimization of ISSR Reaction System for Piper nigrum L.%胡椒ISSR反应体系的建立与优化

    Institute of Scientific and Technical Information of China (English)

    范睿; 郝朝运; 闫林; 黄丽芳; 谭乐和; 杨建峰; 邬华松

    2012-01-01

    通过单因子、双因子实验研究了胡椒ISSR-PCR反应体系中热参数和5个主要成分,即退火温度、循环数、变性时间、退火时间、延伸时间以及Mg2+、dNTPS、引物、模板DNA、TaqDNA聚合酶对扩增结果的影响,建立了适合胡椒ISSR分析的反应体系和扩增程序,即在25μL反应体系中,内含2 mmol/L Mg2+、200μmol/LdNTPS、1×PCR Buffer、2μmol/L引物、100 ng模板、1 U Taq DNA聚合酶。扩增程序为94℃预变性3 min,94℃变性120 s,复性60 s,72℃延伸3 min,循环35个,结束后72℃延伸7 min。这一优化体系的建立为今后利用ISSR标记技术进行胡椒种质鉴定、遗传多样性分析奠定了基础。%Based on the genomic DNA extracted from Piper rtigrum L., this paper presented the effect of the main reaction system elements (Mg2+, dNTPS, primer, template DNA, Taq DNA polymerase) and hot-cycle parameters (annealing temperature, cycles, denaturing time, annealing time and extension time) on ISSR-PCR which were tested by single or dual factor experiment, respectively, to determine its optimal levels. A reaction system and amplified procedure suitable for Piper nigrum L. were established, that is, 25 μL amplification reaction system containing 1 ×PCR Buffer, 2 mmol/L Mg2+, 200μmol/L dNTPS, 2 μmol/L primer, 100 ng DNA and 1 U Taq DNA polymerase. The optimal amp:ified procedure was devised for 3 minutes of predenaturalization at 94~C. 35 cycles of 120 seconds for denaturalization at 94℃, 60 seconds of annealing due to denaturing temperature of different primer, 180 seconds of extension at 72℃ and 7 min of extension at 72℃ in the final cycle. This optimal system laid the standardization program of the identification of Piper nigrum L. and the efficient use of its germplasm resource.

  2. Analysis of Simple Sequence Repeats in Genomes of Rhizobia

    Institute of Scientific and Technical Information of China (English)

    GAO Ya-mei; HAN Yi-qiang; TANG Hui; SUN Dong-mei; WANG Yan-jie; WANG Wei-dong

    2008-01-01

    Simple sequence repeats (SSRs) or microsatellites, as genetic markers, are ubiquitous in genomes of various organisms. The analysis of SSR in rhizobia genome provides useful information for a variety of applications in population genetics of rhizobia. We analyzed the occurrences, relative abundance, and relative density of SSRs, the most common in Bradyrhizobium japonicum, Mesorhizobium loti, and Sinorhizobium meliloti genomes se-quenced in the microorganisms tandem repeats database, and SSRs in the three species genomes were compared with each other. The result showed that there were 1 410, 859, and 638 SSRs in B. japonicum, M. loti, and 5. meliloti genomes, respectively. In the genomes of B. japonicum, M. loti, and 5. meliloti, tetranucleotide, pentanucleotide, and hexanucleotide repeats were more abundant and indicated higher mutation rates in these species. The least abundance was mononucleotide repeat. The SSRs type and distribution were similar among these species.

  3. ISSR标记技术及其在遗传多样性研究中的应用%Inter-Simple Sequence Repeat and Application in the Research of Genetic Diversity

    Institute of Scientific and Technical Information of China (English)

    谢佳燕; 张知彬

    2004-01-01

    ISSR(Inter-Simple Sequence Repeat)技术是在PCR中直接使用微卫星序列进行DNA扩增的一种DNA分子标记.文章主要介绍了ISSR标记的原理、方法、特点及其在遗传多样性研究中的应用.ISSR标记方法具有无需知道任何靶标序列的微卫星背景信息、遗传多态性高、检测快速等特点,在遗传多样性研究中具有广泛的应用前景.

  4. Expansion of protein domain repeats.

    Directory of Open Access Journals (Sweden)

    Asa K Björklund

    2006-08-01

    Full Text Available Many proteins, especially in eukaryotes, contain tandem repeats of several domains from the same family. These repeats have a variety of binding properties and are involved in protein-protein interactions as well as binding to other ligands such as DNA and RNA. The rapid expansion of protein domain repeats is assumed to have evolved through internal tandem duplications. However, the exact mechanisms behind these tandem duplications are not well-understood. Here, we have studied the evolution, function, protein structure, gene structure, and phylogenetic distribution of domain repeats. For this purpose we have assigned Pfam-A domain families to 24 proteomes with more sensitive domain assignments in the repeat regions. These assignments confirmed previous findings that eukaryotes, and in particular vertebrates, contain a much higher fraction of proteins with repeats compared with prokaryotes. The internal sequence similarity in each protein revealed that the domain repeats are often expanded through duplications of several domains at a time, while the duplication of one domain is less common. Many of the repeats appear to have been duplicated in the middle of the repeat region. This is in strong contrast to the evolution of other proteins that mainly works through additions of single domains at either terminus. Further, we found that some domain families show distinct duplication patterns, e.g., nebulin domains have mainly been expanded with a unit of seven domains at a time, while duplications of other domain families involve varying numbers of domains. Finally, no common mechanism for the expansion of all repeats could be detected. We found that the duplication patterns show no dependence on the size of the domains. Further, repeat expansion in some families can possibly be explained by shuffling of exons. However, exon shuffling could not have created all repeats.

  5. Methods for Development of Microsatellite Markers: An Overview

    Directory of Open Access Journals (Sweden)

    Siju SENAN

    2014-03-01

    Full Text Available Microsatellite or Simple Sequence Repeat (SSR markers have evolved to the status of a most versatile and popular genetic marker in a ubiquity of plant systems. Due to their co-dominant, hyper-variable and multiallelic nature, they are the prominent markers of choice for fingerprinting, conservation genetics, plant breeding and phylogenetic studies. Despite its development of a new set of SSR markers for a species remained time consuming and expensive for many years. However, with the recent advancement in genomics, new strategies/protocols are now available for the generation of SSR markers. This review presents an overview on microsatellite markers with a special emphasis on the various strategies used for the development of microsatellite markers

  6. Development and validation of new SSR markers from expressed regions in the garlic genome

    Science.gov (United States)

    Limited number of simple sequence repeat (SSR) markers is available for the genome of garlic (Allium sativum L.) although SSR markers have become one of the most preferred marker systems because they are typically co-dominant, reproducible, cross species transferable and highly polymorphic. In this ...

  7. DWI Repeaters and Non-Repeaters: A Comparison.

    Science.gov (United States)

    Weeber, Stan

    1981-01-01

    Discussed how driving-while-intoxicated (DWI) repeaters differed signigicantly from nonrepeaters on 4 of 23 variables tested. Repeaters were more likely to have zero or two dependent children, attend church frequently, drink occasionally and have one or more arrests for public intoxication. (Author)

  8. To Repeat or Not to Repeat a Course

    Science.gov (United States)

    Armstrong, Michael J.; Biktimirov, Ernest N.

    2013-01-01

    The difficult transition from high school to university means that many students need to repeat (retake) 1 or more of their university courses. The authors examine the performance of students repeating first-year core courses in an undergraduate business program. They used data from university records for 116 students who took a total of 232…

  9. Assessment of genetic diversity in Isabgol (Plantago ovata Forsk ...

    African Journals Online (AJOL)

    sandeep kaswan

    using random amplified polymorphic DNA ... RAPD markers appeared more informative than ISSR in determining the genetic ... Key words: Plantago ovata, molecular marker, RAPD, ISSR, genetic diversity, medicinal plant. .... monomorphic.

  10. Genetic diversity analysis in the Hypericum perforatum populations ...

    African Journals Online (AJOL)

    user

    2014-01-01

    Jan 1, 2014 ... Analysis of molecular variance by ISSR markers indicated that over half of the .... spacer; Rp, resolving power; PIC, polymorphic information content; MI, marker index ..... ISSR 07 revealed 24 monomorphic loci existed in all of.

  11. Molecular Identification and Cultivar Fingerprints of Prunus persica (L.)Batsch Germplasms

    Institute of Scientific and Technical Information of China (English)

    SUN Shu-xia; LI Jing; JIANG Guo-liang; CHEN Dong; XIE Hong-jiang; TU Mei-yan

    2010-01-01

    [Objective]The aim was to study the molecular identification and cultivar fingerprints of Prunus persica(L.)Batsch germplasms.[Method]Sixty peach genotypes,representing China common local cultivars and European samples were screened by microsatellites(simple sequence repeats,SSRs)and Inter-Simple Sequence Repeat(ISSR)markers.[Result]26 reproducible bands were amplified by Nine SSR primers,and 24 of which were polymorphic; 236 bands were amplified by 30 ISSR primers,and 113 of which were polymorphic.31 genotypes were discriminated with 1-3 distinct polymorphic bands generated from the primers ISSR and SSR.Seven cultivar-specific ISSR fragments and two SSR unique alleles obtained from this study were available to be converted into Sequence Characterized Amplified Region(SCAR)markers.The genetic similarity coefficient(GS)estimated from these molecular data averaged were 0.939(ranged from 0.856 to 0.983)for ISSR and0.646(ranged from 0.240 to 1.000)for SSR,respectively.The combined grouping association indicated that most local Chinese peach cultivars and exotic accessions were clustered together.This could be related to the mode of introduction and maintenance of the peach cultivars involving limited foundation germplasm,exchange of cultivars between plantations,and periodic development of new recombinant cultivars following sexual reproduction.[Conclusion]The results obtained in this work would help to improve the conservation,molecular identification and management of peach germplasm in breeding.

  12. Ceramic subsurface marker prototypes

    Energy Technology Data Exchange (ETDEWEB)

    Lukens, C.E. [Rockwell International Corp., Richland, WA (United States). Rockwell Hanford Operations

    1985-05-02

    The client submitted 5 sets of porcelain and stoneware subsurface (radioactive site) marker prototypes (31 markers each set). The following were determined: compressive strength, thermal shock resistance, thermal crazing resistance, alkali resistance, color retention, and chemical resistance.

  13. Estimating genetic diversity and sampling strategy for a wild soybean (Glycine soja) population based on different molecular markers

    Institute of Scientific and Technical Information of China (English)

    CHEN Zhong; ZHAO Ru; GU Senchang; YAN Wen; CHENG Zhou; CHEN Muhong; LU Weifeng; WANG Shuhong; LU Baorong; LU Jun; ZHANG Fan; XIANG Rong; XIAO Shangbin; YAN Pin

    2006-01-01

    Genetic diversity is the basic and most important component of biodiversity. It is essential for the effective conservation and utilization of genetic resources to accurately estimate genetic diversity of the targeted species and populations. This paper reports analyses of genetic diversity of a wild soybean population using three molecular marker technologies (AFLP, ISSR and SSR), and computer simulation studies of randomly selected subsets with different sample size (5-90 individuals) drawn 50 times from a total of 100 wild soybean individuals. The variation patterns of genetic diversity indices, including expected heterozygosity (He), Shannon diversity index (/), and percentage of polymorphic loci (P), were analyzed to evaluate changes of genetic diversity associated with the increase of individuals in each subset. The results demonstrated that (1) values of genetic diversity indices of the same wild soybean population were considerably different when estimated by different molecular marker techniques; (2) genetic diversity indices obtained from subsets with different sample sizes also diverged considerably; (3) P values were relatively more reliable for comparing genetic diversity detected by different molecular marker techniques; and (4) different diversity indices reached 90% of the total genetic diversity of the soybean population quite differently in terms of the sample size (number of individuals) analyzed.When using the P value as a determinator, 30-40individuals could capture over 90% of the total genetic diversity of the wild soybean population. Results from this study provide a strong scientific basis for estimating genetic diversity and for strategic conservation of plant species.

  14. Nifty Nines and Repeating Decimals

    Science.gov (United States)

    Brown, Scott A.

    2016-01-01

    The traditional technique for converting repeating decimals to common fractions can be found in nearly every algebra textbook that has been published, as well as in many precalculus texts. However, students generally encounter repeating decimal numerals earlier than high school when they study rational numbers in prealgebra classes. Therefore, how…

  15. Nifty Nines and Repeating Decimals

    Science.gov (United States)

    Brown, Scott A.

    2016-01-01

    The traditional technique for converting repeating decimals to common fractions can be found in nearly every algebra textbook that has been published, as well as in many precalculus texts. However, students generally encounter repeating decimal numerals earlier than high school when they study rational numbers in prealgebra classes. Therefore, how…

  16. All-photonic quantum repeaters

    Science.gov (United States)

    Azuma, Koji; Tamaki, Kiyoshi; Lo, Hoi-Kwong

    2015-01-01

    Quantum communication holds promise for unconditionally secure transmission of secret messages and faithful transfer of unknown quantum states. Photons appear to be the medium of choice for quantum communication. Owing to photon losses, robust quantum communication over long lossy channels requires quantum repeaters. It is widely believed that a necessary and highly demanding requirement for quantum repeaters is the existence of matter quantum memories. Here we show that such a requirement is, in fact, unnecessary by introducing the concept of all-photonic quantum repeaters based on flying qubits. In particular, we present a protocol based on photonic cluster-state machine guns and a loss-tolerant measurement equipped with local high-speed active feedforwards. We show that, with such all-photonic quantum repeaters, the communication efficiency scales polynomially with the channel distance. Our result paves a new route towards quantum repeaters with efficient single-photon sources rather than matter quantum memories. PMID:25873153

  17. Expression and its significance of stem cells marker leucine-rich repeat containing G protein coupled receptor 5 gene in human colorectal cancer%人结直肠癌干细胞标志物富含亮氨酸重复单位的G蛋白耦联受体5基因的表达及意义

    Institute of Scientific and Technical Information of China (English)

    孙艳; 盛春华; 文大成; 李玉林; 迟宝荣

    2013-01-01

    目的 探讨人结直肠癌组织及外周血中干细胞标志物富含亮氨酸重复单位的G蛋白耦联受体5(lgr5)基因的表达及其与临床病理特征间的关系.方法 采用SYBR Green实时定量PCR方法检测27份结直肠癌组织及配对的正常组织中、1 7例患者及8名健康对照者外周血中lgr5mRNA的表达.应用Wilcoxon秩和检验分析lgr5 mRNA在不同组织间、临床病理参数之间的表达差异.结果 结直肠癌组织中lgr5 mRNA表达水平为1.000(0.012,496.353),高于配对正常组织的0.147(0.004,73.002),差异有统计学意义(Z=8.029,P<0.01).结直肠癌患者外周血中lgr5mRNA表达水平为0.742(0.077,456.566),高于健康对照组的0.104(0.034,0.274),差异有统计学意义(Z=2.048,P<0.05).结直肠癌组织lgr5 mRNA在不同性别、年龄、肿瘤原发部位、肿瘤大小、组织学类型组间表达差异均无统计学意义(P均>0.05),但有淋巴结转移组lgr5 mRNA表达高于无淋巴结转移组,差异有统计学意义(Z=2.066,P<0.05).结论 lgr5在结直肠癌组织及外周血中的表达上调可能参与了结直肠癌的生长及转移.%Objective To investigate the expression of stem cell marker leucine-rich repeat containing G protein coupled receptor 5 (lgr5) gene in human colorectal cancer tissues and peripheral blood and its correlation with clinical pathological characteristics.Methods The expression of lgr5 at mRNA level was detected by SYBR Green quantitative real-time polymerase chain reaction (PCR) in 27 human colorectal cancer tissues and corresponding non-cancerous tissues as well as in peripheral blood of 17 patients and eight healthy controls.The differences of lgr5 mRNA expression in different tissues and clinical pathology parameters were analyzed by Wilcoxon test.Results The expression of lgr5 at mRNA level in colorectal cancer tissues was 1.000 (0.012,496.353),which was higher than that of corresponding non-cancerous tissues 0.147 (0.004,73.002),the

  18. Morphological and comparative genomic analyses of pathogenic and non-pathogenic Fusarium solani isolated from Dalbergia sissoo.

    Science.gov (United States)

    Arif, M; Zaidi, N W; Haq, Q M R; Singh, Y P; Taj, G; Kar, C S; Singh, U S

    2015-06-01

    Sissoo or shisham (Dalbergia sissoo Roxb.) is one of the finest wood of South Asia. Fusarium solani is a causal organism of sissoo wilt, decline, or dieback. It is also a potential causal organism associated with other valuable tree species. Thirty-eight Fusarium isolates including 24 F. solani and 14 Fusarium sp., were obtained in 2005 from different geographical locations in India. All 38 (18 pathogenic and 20 non-pathogenic) isolates were characterized for genomic analysis, growth behaviour, pigmentation and sensitivity to carbendazim. Based on growth pattern, growth rate, pigmentation and sensitivity to carbendazim, all 38 isolates showed a wide range of variability, but no correlation with pathogenicity or geographical distribution. Three techniques were used for comparative genomic analysis: random amplified polymorphic DNA (RAPD); inter simple sequence repeats (ISSR); and simple sequence repeats (SSR). A total of 90 primers targeting different genome regions resulted a total of 1159 loci with an average of 12.88 loci per primer. These primers showed high genomic variability among the isolates. The maximum loci (14.64) per primer were obtained with RAPD. The total variation of the first five principal components for RAPD, ISSR, SSR and combined analysis were estimated as 47.42, 48.21, 46.30 and 46.78 %, respectively. Among the molecular markers, highest Pearson correlation value (r = 0.957) was recorded with combination of RAPD and SSR followed by RAPD and ISSR (r = 0.952), and SSR and ISSR (r = 0.942). The combination of these markers would be similarly effective as single marker system i.e. RAPD, ISSR and SSR. Based on polymorphic information content (PIC = 0.619) and highest coefficient (r = 0.995), RAPD was found to be the most efficient marker system compared to ISSR and SSR. This study will assist in understanding the population biology of wilt causing phytopathogen, F. solani, and in assisting with integrated disease management measures.

  19. Oxygen uptake during repeated-sprint exercise.

    Science.gov (United States)

    McGawley, Kerry; Bishop, David J

    2015-03-01

    Repeated-sprint ability appears to be influenced by oxidative metabolism, with reductions in fatigue and improved sprint times related to markers of aerobic fitness. The aim of the current study was to measure the oxygen uptake (VO₂) during the first and last sprints during two, 5 × 6-s repeated-sprint bouts. Cross-sectional study. Eight female soccer players performed two, consecutive, 5 × 6-s maximal sprint bouts (B1 and B2) on five separate occasions, in order to identify the minimum time (trec) required to recover total work done (Wtot) in B1. On a sixth occasion, expired air was collected during the first and last sprint of B1 and B2, which were separated by trec. The trec was 10.9 ± 1.1 min. The VO₂ during the first sprint was significantly less than the last sprint in each bout (psprint (measured in kJ) was significantly related to VO₂max in both B1 (r=0.81, p=0.015) and B2 (r=0.93, p=0.001). In addition, the VO₂ attained in the final sprint was not significantly different from VO₂max in B1 (p=0.284) or B2 (p=0.448). The current study shows that the VO₂ increases from the first to the last of 5 × 6-s sprints and that VO₂max may be a limiting factor to performance in latter sprints. Increasing V˙O₂max in team-sport athletes may enable increased aerobic energy delivery, and consequently work done, during a bout of repeated sprints. Copyright © 2014 Sports Medicine Australia. Published by Elsevier Ltd. All rights reserved.

  20. Simple sequence repeat map of the sunflower genome.

    Science.gov (United States)

    Tang, S.; Yu, J.-K.; Slabaugh, B.; Shintani, K.; Knapp, J.

    2002-12-01

    Several independent molecular genetic linkage maps of varying density and completeness have been constructed for cultivated sunflower ( Helianthus annuus L.). Because of the dearth of sequence and probe-specific DNA markers in the public domain, the various genetic maps of sunflower have not been integrated and a single reference map has not emerged. Moreover, comparisons between maps have been confounded by multiple linkage group nomenclatures and the lack of common DNA markers. The goal of the present research was to construct a dense molecular genetic linkage map for sunflower using simple sequence repeat (SSR) markers. First, 879 SSR markers were developed by identifying 1,093 unique SSR sequences in the DNA sequences of 2,033 clones isolated from genomic DNA libraries enriched for (AC)(n) or (AG)(n) and screening 1,000 SSR primer pairs; 579 of the newly developed SSR markers (65.9% of the total) were polymorphic among four elite inbred lines (RHA280, RHA801, PHA and PHB). The genetic map was constructed using 94 RHA280 x RHA801 F(7) recombinant inbred lines (RILs) and 408 polymorphic SSR markers (462 SSR marker loci segregated in the mapping population). Of the latter, 459 coalesced into 17 linkage groups presumably corresponding to the 17 chromosomes in the haploid sunflower genome ( x = 17). The map was 1,368.3-cM long and had a mean density of 3.1 cM per locus. The SSR markers described herein supply a critical mass of DNA markers for constructing genetic maps of sunflower and create the basis for unifying and cross-referencing the multitude of genetic maps developed for wild and cultivated sunflowers.

  1. Profile of candidate microsatellite markers in Sebastiscus marmoratus using 454 pyrosequencing

    Science.gov (United States)

    Song, Na; Chen, Muyan; Gao, Tianxiang; Yanagimoto, Takashi

    2017-01-01

    Sebastiscus marmoratus is an important sedentary ovoviparous fish distributed in near-shore coastal waters from the coast of China to Japan. Candidate S. marmoratus microsatellite markers were developed in the present study using 454 pyrosequencing, and the marker profile was analyzed. A total of 2 000 000 raw sequence reads were assembled to reduce redundancy. Among them, 1 043 dinucleotide, 925 trinucleotide, 692 tetranucleotide, and 315 pentanucleotide repeats were detected. AC repeats were the most frequent motifs among the dinucleotide repeats, and AAT was the most abundant among the trinucleotide repeats. AAAT, ATAG, and ATCC were the three most common tetranucleotide motifs, and AAGAT and AATAT were the most dominant pentanucleotide motifs. The greatest numbers of loci and potentially amplifiable loci were found in dinucleotide repeats, whereas trinucleotide repeats had the fewest. In summary, a wide range of candidate microsatellite markers were identified in the present study using a rapid and efficient 454 pyrosequencing approach.

  2. Analysis of Genetic Polymorphic SSR Markers in Germplasm Resources of the Natural Colored Cotton

    Institute of Scientific and Technical Information of China (English)

    WANG Ju-qin; LI Fu-zhen; QIU Xin-mian; BAO Li-sheng; LU Yan-ting

    2008-01-01

    @@ Short sequence repeats (microsatellite,SSR) and expressed sequence tags-SSR (EST-SSR) markers were employed to analyze the genetic diversity of natural colored cotton varieties.About 490 pairs of SSR markers spanning the 26 chromosomes were selected from the cotton microsatellite database,they were composed of the NAU,BNL,MUSS,and CIR markers,and there was one marker every 5 cM on average.

  3. Congruence between morphological and molecular markers inferred from the analysis of the intra-morphotype genetic diversity and the spatial structure of Oxalis tuberosa Mol.

    Science.gov (United States)

    Pissard, Audrey; Arbizu, Carlos; Ghislain, Marc; Faux, Anne-Michèle; Paulet, Sébastien; Bertin, Pierre

    2008-01-01

    Oxalis tuberosa is an important crop cultivated in the highest Andean zones. A germplasm collection is maintained ex situ by CIP, which has developed a morphological markers system to classify the accessions into morphotypes, i.e. groups of morphologically identical accessions. However, their genetic uniformity is currently unknown. The ISSR technique was used in two experiments to determine the relationships between both morphological and molecular markers systems. The intra-morphotype genetic diversity, the spatial structures of the diversity and the congruence between both markers systems were determined. In the first experience, 44 accessions representing five morphotypes, clearly distinct from each other, were analyzed. At the molecular level, the accessions exactly clustered according to their morphotypes. However, a genetic variability was observed inside each morphotype. In the second experiment, 34 accessions gradually differing from each other on morphological base were analyzed. The morphological clustering showed no geographical structure. On the opposite, the molecular analysis showed that the genetic structure was slightly related to the collection site. The correlation between both markers systems was weak but significant. The lack of perfect congruence between morphological and molecular data suggests that the morphological system may be useful for the morphotypes management but is not appropriate to study the genetic structure of the oca. The spatial structure of the genetic diversity can be related to the evolution of the species and the discordance between the morphological and molecular structures may result from similar selection pressures at different places leading to similar forms with a different genetic background.

  4. Analysis of repeated measures data

    CERN Document Server

    Islam, M Ataharul

    2017-01-01

    This book presents a broad range of statistical techniques to address emerging needs in the field of repeated measures. It also provides a comprehensive overview of extensions of generalized linear models for the bivariate exponential family of distributions, which represent a new development in analysing repeated measures data. The demand for statistical models for correlated outcomes has grown rapidly recently, mainly due to presence of two types of underlying associations: associations between outcomes, and associations between explanatory variables and outcomes. The book systematically addresses key problems arising in the modelling of repeated measures data, bearing in mind those factors that play a major role in estimating the underlying relationships between covariates and outcome variables for correlated outcome data. In addition, it presents new approaches to addressing current challenges in the field of repeated measures and models based on conditional and joint probabilities. Markov models of first...

  5. Molecular marker databases.

    Science.gov (United States)

    Lai, Kaitao; Lorenc, Michał Tadeusz; Edwards, David

    2015-01-01

    The detection and analysis of genetic variation plays an important role in plant breeding and this role is increasing with the continued development of genome sequencing technologies. Molecular genetic markers are important tools to characterize genetic variation and assist with genomic breeding. Processing and storing the growing abundance of molecular marker data being produced requires the development of specific bioinformatics tools and advanced databases. Molecular marker databases range from species specific through to organism wide and often host a variety of additional related genetic, genomic, or phenotypic information. In this chapter, we will present some of the features of plant molecular genetic marker databases, highlight the various types of marker resources, and predict the potential future direction of crop marker databases.

  6. Diversity in needle morphology and genetic markers in a marginal Abies cephalonica (Pinaceae population

    Directory of Open Access Journals (Sweden)

    Aristotelis C. Papageorgiou

    2015-12-01

    Full Text Available Differences in needle traits of coniferous tree species are considered as the combined result of direct environmental pressure and specific genetic adaptations. In this study, diversity and differentiation within and among four Abies cephalonica subpopulations of a marginal population on Mt. Parnitha - Greece, were estimated using needle morphological traits and gene markers. We tested the connection of morphological variability patterns of light and shade needles with possible adaptation strategies and genetic diversity. Six morphological characteristics were used for the description of both light and shade needles at 100 trees, describing needle size and shape, stomatal density and needle position on the twigs. Additionally, six RAPD and three ISSR markers were applied on DNA from the same trees. Light needles were significantly different than shade needles, in all traits measured, apparently following a different light harvesting strategy. All four subpopulations exhibited high genetic diversity and the differentiation among them was relatively low. Differences among populations in light needles seemed to depend on light exposure and aspect. In shade needles, the four subpopulations seemed to deviate stronger from each other and express a rather geographic pattern, similarly to the genetic markers. Two of the subpopulations studied were lost during a wildfire, two years after sampling. Although the subpopulations burnt were most diverse and most differentiated, we expect a large part of the total genetic diversity of the burnt trees to still exist in the surviving subpopulations, since gene flow must have been effective in keeping all subpopulations connected.

  7. 棉花遗传作图用SSR标记的开发%Development of SSR Markers towards Genetic Mapping in Cotton (GossyPium hirsutum L. )

    Institute of Scientific and Technical Information of China (English)

    Siva P. KUMPATLA; Erin C. HORNE; Manali R. SHAH; Manju GUPTA; Steven A. THOMPSON

    2002-01-01

    @@ Availability of informative molecular markers is a prerequisite for genetic mapping and marker assisted selection projects. Micro-satellites or Simple Sequence Repeat (SSR) markers are PCR-based and currently the most widely used marker system in the plant molecular genetics community due to their high degree of polymorphism, random distribution throughout the genome and their suitability for high throughput genotyping formats.

  8. A Set of SCAR Markers Efficiently Differentiating Hybrid Rice

    Institute of Scientific and Technical Information of China (English)

    LI Shu-jing; XIE Hong-wei; QIAN Ming-juan; CHEN Guang-hui; LI Shao-qing; ZHU Ying-guo

    2012-01-01

    Molecular markers have been widely used in crop genetic improvement,seed test and genetic mapping.Of which,sequence characterized amplified region (SCAR) markers are particularly popular for its diversity,stable reproducibility,and suitability for analyzing large number of samples.In this study,500 random amplified polymorphic DNA (RAPD) primers were tested,and a set of SCAR markers comprising 37 pairs of loci-specific primers were developed from the DNA fragments ranging from 300 to 1000 bp which correspond to the stable,distinctive RAPD banding patterns.Using these SCAR markers,59 hybrid rice combinations were assessed and distinguished into 58 subgroups at the similarity coefficient of 0.97 in a genetic clustering tree based on the allele diversities of the SCAR markers.Furthermore,13 hybrid rice combinations were reassayed with 40 randomly selected simple sequence repeat (SSR) markers to evaluate the effectiveness of these SCAR markers.SSR markers produced similar results to SCAR markers as the 13 hybrid rice combinations were completely separated at the similarity coefficient of 0.91 in the clustering tree established from SSR patterns.Taken together,SCAR markers prove to be effective tools for identifying and differentiating hybrid rice combinations.

  9. A Set of SCAR Markers Efficiently Differentiating Hybrid Rice

    Directory of Open Access Journals (Sweden)

    Shu-jing LI

    2012-03-01

    Full Text Available Molecular markers have been widely used in crop genetic improvement, seed test and genetic mapping. Of which, sequence characterized amplified region (SCAR markers are particularly popular for its diversity, stable reproducibility, and suitability for analyzing large number of samples. In this study, 500 random amplified polymorphic DNA (RAPD primers were tested, and a set of SCAR markers comprising 37 pairs of loci-specific primers were developed from the DNA fragments ranging from 300 to 1000 bp which correspond to the stable, distinctive RAPD banding patterns. Using these SCAR markers, 59 hybrid rice combinations were assessed and distinguished into 58 subgroups at the similarity coefficient of 0.97 in a genetic clustering tree based on the allele diversities of the SCAR markers. Furthermore, 13 hybrid rice combinations were reassayed with 40 randomly selected simple sequence repeat (SSR markers to evaluate the effectiveness of these SCAR markers. SSR markers produced similar results to SCAR markers as the 13 hybrid rice combinations were completely separated at the similarity coefficient of 0.91 in the clustering tree established from SSR patterns. Taken together, SCAR markers prove to be effective tools for identifying and differentiating hybrid rice combinations.

  10. Limitations on quantum key repeaters.

    Science.gov (United States)

    Bäuml, Stefan; Christandl, Matthias; Horodecki, Karol; Winter, Andreas

    2015-04-23

    A major application of quantum communication is the distribution of entangled particles for use in quantum key distribution. Owing to noise in the communication line, quantum key distribution is, in practice, limited to a distance of a few hundred kilometres, and can only be extended to longer distances by use of a quantum repeater, a device that performs entanglement distillation and quantum teleportation. The existence of noisy entangled states that are undistillable but nevertheless useful for quantum key distribution raises the question of the feasibility of a quantum key repeater, which would work beyond the limits of entanglement distillation, hence possibly tolerating higher noise levels than existing protocols. Here we exhibit fundamental limits on such a device in the form of bounds on the rate at which it may extract secure key. As a consequence, we give examples of states suitable for quantum key distribution but unsuitable for the most general quantum key repeater protocol.

  11. Hysteresis of magnetostructural transitions: Repeatable and non-repeatable processes

    Energy Technology Data Exchange (ETDEWEB)

    Provenzano, Virgil [National Institute of Standards and Technology, Gaithersburg, MD 20899 (United States); Della Torre, Edward; Bennett, Lawrence H. [Department of Electrical and Computer Engineering, The George Washington University, Washington, DC 20052 (United States); ElBidweihy, Hatem, E-mail: Hatem@gwmail.gwu.edu [Department of Electrical and Computer Engineering, The George Washington University, Washington, DC 20052 (United States)

    2014-02-15

    The Gd{sub 5}Ge{sub 2}Si{sub 2} alloy and the off-stoichiometric Ni{sub 50}Mn{sub 35}In{sub 15} Heusler alloy belong to a special class of metallic materials that exhibit first-order magnetostructural transitions near room temperature. The magnetic properties of this class of materials have been extensively studied due to their interesting magnetic behavior and their potential for a number of technological applications such as refrigerants for near-room-temperature magnetic refrigeration. The thermally driven first-order transitions in these materials can be field-induced in the reverse order by applying a strong enough field. The field-induced transitions are typically accompanied by the presence of large magnetic hysteresis, the characteristics of which are a complicated function of temperature, field, and magneto-thermal history. In this study we show that the virgin curve, the major loop, and sequentially measured MH loops are the results of both repeatable and non-repeatable processes, in which the starting magnetostructural state, prior to the cycling of field, plays a major role. Using the Gd{sub 5}Ge{sub 2}Si{sub 2} and Ni{sub 50}Mn{sub 35}In{sub 15} alloys, as model materials, we show that a starting single phase state results in fully repeatable processes and large magnetic hysteresis, whereas a mixed phase starting state results in non-repeatable processes and smaller hysteresis.

  12. Physiological consequences of repeated exposures to conditioned fear.

    Science.gov (United States)

    Thompson, Robert S; Strong, Paul V; Fleshner, Monika

    2012-06-01

    Activation of the stress response evokes a cascade of physiological reactions that may be detrimental when repeated or chronic, and when triggered after exposure to psychological/emotional stressors. Investigation of the physiological mechanisms responsible for the health damaging effects requires animal paradigms that repeatedly evoke a response to psychological/emotional stressors. To this end, adult male Sprague Dawley rats were repeatedly exposed (2X per day for 20 days) to a context that they were conditioned to fear (conditioned fear test, CFT). Repeated exposure to CFT produced body weight loss, adrenal hypertrophy, thymic involution, and basal corticosterone elevation. In vivo biotelemetry measures revealed that CFT evokes sympathetic nervous system driven increases in heart rate (HR), mean arterial pressure (MAP), and core body temperature. Extinction of behavioral (freezing) and physiological responses to CFT was prevented using minimal reinstatement footshock. MAP responses to the CFT did not diminish across 20 days of exposure. In contrast, HR and cardiac contractility responses declined by day 15, suggesting a shift toward vascular-dominated MAP (a pre-clinical marker of CV dysfunction). Flattened diurnal rhythms, common to stress-related mood/anxiety disorders, were found for most physiological measures. Thus, repeated CFT produces adaptations indicative of the health damaging effects of psychological/emotional stress.

  13. Physiological Consequences of Repeated Exposures to Conditioned Fear

    Directory of Open Access Journals (Sweden)

    Robert S. Thompson

    2012-05-01

    Full Text Available Activation of the stress response evokes a cascade of physiological reactions that may be detrimental when repeated or chronic, and when triggered after exposure to psychological/emotional stressors. Investigation of the physiological mechanisms responsible for the health damaging effects requires animal paradigms that repeatedly evoke a response to psychological/emotional stressors. To this end, adult male Sprague Dawley rats were repeatedly exposed (2X per day for 20 days to a context that they were conditioned to fear (conditioned fear test, CFT. Repeated exposure to CFT produced body weight loss, adrenal hypertrophy, thymic involution, and basal corticosterone elevation. In vivo biotelemetry measures revealed that CFT evokes sympathetic nervous system driven increases in heart rate (HR, mean arterial pressure (MAP, and core body temperature. Extinction of behavioral (freezing and physiological responses to CFT was prevented using minimal reinstatement footshock. MAP responses to the CFT did not diminish across 20 days of exposure. In contrast, HR and cardiac contractility responses declined by day 15, suggesting a shift toward vascular-dominated MAP (a pre-clinical marker of CV dysfunction. Flattened diurnal rhythms, common to stress-related mood/anxiety disorders, were found for most physiological measures. Thus, repeated CFT produces adaptations indicative of the health damaging effects of psychological/emotional stress.

  14. Polymorphic microsatellite markers in Euryale ferox Salisb. (Nymphaeaceae).

    Science.gov (United States)

    Quan, Zhiwu; Pan, Lei; Ke, Weidong; Ding, Yi

    2009-01-01

    Eleven polymorphic microsatellite markers were isolated and identified in the aquatic plant Euryale ferox Salisb. (Nymphaeaceae). This species, which belongs to basal Magnoliophyta, reproduces sexually. All of these 11 microsatellite markers yielded 25 alleles in a survey of a wild population of 34 individuals. Two or three alleles per locus were detected, with expected heterozygosity ranging from 0.056 to 0.634 and observed heterozygosity from 0.000 to 0.088. These simple sequence repeat markers will be useful for evaluating the genetic structure of the E. ferox population in the future.

  15. Genomic sequencing and microsatellite marker development for Boswellia papyrifera, an economically important but threatened tree native to dry tropical forests

    NARCIS (Netherlands)

    Addisalem, A.B.; Esselink, G.; Bongers, F.; Smulders, M.J.M.

    2015-01-01

    Microsatellite (or simple sequence repeat, SSR) markers are highly informative DNA markers often used in conservation genetic research. Next-generation sequencing enables efficient development of large numbers of SSR markers at lower costs. Boswellia papyrifera is an economically important tree spec

  16. Embryonic Stem Cell Markers

    Directory of Open Access Journals (Sweden)

    Lan Ma

    2012-05-01

    Full Text Available Embryonic stem cell (ESC markers are molecules specifically expressed in ES cells. Understanding of the functions of these markers is critical for characterization and elucidation for the mechanism of ESC pluripotent maintenance and self-renewal, therefore helping to accelerate the clinical application of ES cells. Unfortunately, different cell types can share single or sometimes multiple markers; thus the main obstacle in the clinical application of ESC is to purify ES cells from other types of cells, especially tumor cells. Currently, the marker-based flow cytometry (FCM technique and magnetic cell sorting (MACS are the most effective cell isolating methods, and a detailed maker list will help to initially identify, as well as isolate ESCs using these methods. In the current review, we discuss a wide range of cell surface and generic molecular markers that are indicative of the undifferentiated ESCs. Other types of molecules, such as lectins and peptides, which bind to ESC via affinity and specificity, are also summarized. In addition, we review several markers that overlap with tumor stem cells (TSCs, which suggest that uncertainty still exists regarding the benefits of using these markers alone or in various combinations when identifying and isolating cells.

  17. Marker development in ornamental plants

    NARCIS (Netherlands)

    Heusden, van A.W.; Arens, P.

    2009-01-01

    Development of markers for a new crop or development of additional markers for a crop where markers have been developed in the past raises the question of the intended use of the markers. Depending on the different objectives in mind one marker type may be better suited then another. In general one

  18. EAMJ Dec. Repeatability.indd

    African Journals Online (AJOL)

    2008-12-12

    Dec 12, 2008 ... Results:Kappa values for four-week repeatability for the wheeze and asthma questions were 0.61 ... for logistic, cultural and ethical reasons, to use ... individual with baseline forced expiratory volume in .... period is likely to also include the effects of true ... data, the writing of the manuscript or the decision.

  19. The use of ISSR markers for species determination and a genetic study of the invasive lionfish in Guanahacabibes, Cuba/Uso de marcadores ISSR para la determinación de especies y estudios genéticos del pez león, especie invasora en Guanahacabibes, Cuba

    National Research Council Canada - National Science Library

    Elizabeth Labastida; Dorka Cobián; Yann Hénaut; María del Carmen García-Rivas; Pedro P Chevalier; Salima Machkour-M'Rabet

    2015-01-01

      The red lionfish (Pterois volitans) and devil fire-fish (Pterois miles) are invasive species that pose a threat to the biodiversity and stability of coral reefs in the Western Atlantic, Gulf of Mexico and Caribbean Sea...

  20. Development and assessment of DArT markers in triticale.

    Science.gov (United States)

    Badea, A; Eudes, F; Salmon, D; Tuvesson, S; Vrolijk, A; Larsson, C-T; Caig, V; Huttner, E; Kilian, A; Laroche, André

    2011-05-01

    Triticale (X Triticosecale Wittm.) is a hybrid derived by crossing wheat (Triticum sp.) and rye (Secale sp.). Till date, only a limited number of simple sequence repeat (SSRs) markers have been used in triticale molecular analyses and there is a need to identify dedicated high-throughput molecular markers to better exploit this crop. The objective of this study was to develop and evaluate diversity arrays technology (DArT) markers in triticale. DArT marker technology offers a high level of multiplexing. Development of new markers from triticale accessions was combined with mining the large collection of previously developed markers in rye and wheat. Three genotyping arrays were used to analyze a collection of 144 triticale accessions. The polymorphism level ranged from 8.6 to 23.8% for wheat and rye DArT markers, respectively. Among the polymorphic markers, rye markers were the most abundant (3,109) followed by wheat (2,214) and triticale (719). The mean polymorphism information content values were 0.34 for rye DArT markers and 0.37 for those from triticale and wheat. High correlation was observed between similarity matrices derived from rye, triticale, wheat and combined marker sets, as well as for the cophenetic values matrices. Cluster analysis revealed genetic relationships among the accessions consistent with the agronomic and pedigree information available. The newly developed triticale DArT markers as well as those originated from rye and wheat provide high quality markers that can be used for diversity analyses and might be exploited in a range of molecular breeding and genomics applications in triticale.

  1. Relationship of 17 Rosa Plants Detected by Morphology and ISSR Analysis%17份蔷薇属植物的亲缘关系的形态学和ISSR分析

    Institute of Scientific and Technical Information of China (English)

    杨帆; 曾丽; 叶康; 赵子刚; 张嫔; 龚霄雯; 尹勤; 孙强

    2011-01-01

    The genetic relationship of 17 Rosa plants is analyzed by ISSR and morphological data. NTSYS2.1 is used in clustering analysis. The results showed: 13 primers were selected to perform ISSR -PCR. A total of 479 DNA bands was obtained, of which 221 bands had polymorphism, the percentage of polymorphic bands was 0.46. All these plants could be divided into three groups by morphological data: the first group was consisted of Rosa rugosa ' Bao Dao' , Rosa chinensis ' Sophie' s perpetral' , Rosa chinensis ' Comtesse dll cayla' , Rosa hybrid tea ' Sophy' s Rose' , Rosa hybrid climbingroses ' Ten Shen Qi' , Rosa chinensis ' Da Fu Gui' , Rose hybrid bracteata ' Mermaid' , Rosa chinensis ' Jin Ou Fan Lv' , Rosa chinensis ' Jin Fen Lian' and Rosa chinensis ' Yi Ji Fen'; the second group was consisted of Rosa chinensis ' Viridiflora' , Rosa chinensis ' Hu Zhong Yue' , Rosa chinensis ' Yu Shi Zhuang' , Rosa hybrid tea ' Yankee Dodle' , Rosa chinensis ' Si Mian Jing' and Rosa chinensis ' Pu Fu Hong'; the third group was consisted of Rosa gallica ' Versicolor'. All these plants can be divided into four groups by ISSR cluster analysis. The first group was consisted of Rosa rugosa ' Bao Dao'; the second group was consisted of Rosa chinensis ' Viridiflora', Rosa chinensis ' Pu Fu Hong', Rosa chinensis ' Comtesse dll cayla' , Rose hybrid bracteata ' Mermaid' , Rosa chinensis ' Si Mian Jing' , Rosa chinensis ' Hu Zhong Yue' and Rosa chinensis ' Yu Shi Zhuang'; the third group was consisted of Rosa hybrid tea ' Sophy' s Rose' and Rosa hy brid tea ' Yankee Dodle'; and the fourth group was consisted of Rosa gallica ' Versicolor' , Rosa chinensis ' Jin Ou Fan Lv' , Rosa hybrid climbingroses ' Ten Shen Qi' , Rosa chinensis ' Sophie' s perpetral' , Rosa chinensis ' Yi Ji Fen' , Rosa chinensis ' Da Fu Gui' and Rosa chinensis ' Jin Fen Lian'. The results of morphologic cluster analysis and ISSR cluster analysis were similar.%利用ISSR分子标记结合形态学指标对17份蔷薇属植物

  2. A test of mink microsatellite markers in the ferret

    DEFF Research Database (Denmark)

    Anistoroaei, Razvan Marian; Christensen, Knud

    2006-01-01

    Short tandem repeats are a source of highly polymorphic markers in mammalian genomes. Genetic variations at these hypervariable loci is extensively used for linkage analysis and to identify individuals, and is very useful for interpopulation and interspecies studies. Fifty-nine microsatellite...

  3. Association between a Tetranucleotide Repeat Polymorphism of SPAG16 Gene and Cataract in Male Children

    Directory of Open Access Journals (Sweden)

    Shipra Mehra

    2013-01-01

    Full Text Available Purpose. Studies involving genotyping of STR markers at 2q34 have repeatedly found the region to host the disease haplotype for pediatric cataract. Present study investigated the association of D2S2944 marker, in sperm associated antigen 16 (SPAG16 gene and rs2289917 polymorphism, in γ-crystallin B gene, with childhood cataract. Methods. 97 pediatric cataract cases and 110 children with no ocular defects were examined for tetranucleotide repeat marker/SNP using PCR-SSLP/RFLP techniques. Polymorphisms were assessed for association using contingency tables and linkage disequilibrium among alleles of the markers was estimated. Energy-optimization program predicted the secondary structure models of repeats of D2S2944. Results. Seven alleles of D2S2944, with 9–15 “GATA” repeats, were observed. Frequency of the longer allele of D2S2944, ≥(GATA13 repeats, was 0.73 in cases and 0.56 in controls (P=0.0123. Male children bearing ≥(GATA13 repeats showed >3-fold higher risk for cataract (CI95% = 1.43–7.00, P=0.0043, Pc=0.0086 as compared to female children (OR=1.19, CI95% = 0.49–2.92, P=0.70. Cases with haplotype—≥(GATA13 of D2S2944 and “C” allele rs2289917—have a higher risk for pediatric cataract (OR=2.952, CI95% = 1.595~5.463, P=0.000453. >(GATA13 repeats formed energetically more favorable stem-loop structure. Conclusion. Intragenic microsatellite repeat expansion in SPAG16 gene increases predisposition to pediatric cataract by probably interfering posttranscriptional events and affecting the expression of adjacent lens transparency gene/s in a gender bias manner.

  4. VT Roadside Historic Markers

    Data.gov (United States)

    Vermont Center for Geographic Information — Roadside Historic Site Marker program has proven an effective way to commemorate Vermont’s many people, events, and places of regional, statewide, or national...

  5. Fiducial Marker Placement

    Science.gov (United States)

    ... Media Computed Tomography (CT) - Body General Ultrasound Ultrasound - Prostate Introduction to Cancer Therapy (Radiation Oncology) Proton Therapy Stereotactic Radiosurgery (SRS) and Stereotactic Body Radiotherapy (SBRT) Images related to Fiducial Marker Placement Sponsored by ...

  6. Marker development in ornamental plants

    OpenAIRE

    2009-01-01

    Development of markers for a new crop or development of additional markers for a crop where markers have been developed in the past raises the question of the intended use of the markers. Depending on the different objectives in mind one marker type may be better suited then another. In general one can think of two main objectives for the use of markers; variety identification and breeding applications. In view of recent developments in molecular genetics, and sequencing technologies in parti...

  7. Directionality switchable gain stabilized linear repeater

    Science.gov (United States)

    Ota, Takayuki; Ohmachi, Tadashi; Aida, Kazuo

    2004-10-01

    We propose a new approach to realize a bidirectional linear repeater suitable for future optical internet networks and fault location in repeater chain with OTDR. The proposed approach is the linear repeater of simple configuration whose directionality is rearranged dynamically by electrical control signal. The repeater is composed of a magneto-optical switch, a circulator, a dynamically gain stabilized unidirectional EDFA, and control circuits. The repeater directionality is rearranged as fast as 0.1ms by an electrical control pulse. It is experimentally confirmed that OTDR with the directionality switchable repeater is feasible for repeater chain. The detailed design and performance of the repeater are also discussed, including the multi-pass interference (MPI) which may arise in the proposed repeater, the effect of the MPI on SNR degradation of the repeater chain and the feed-forward EDFA gain control circuit.

  8. Selection of microsatellite markers for bladder cancer diagnosis without the need for corresponding blood

    DEFF Research Database (Denmark)

    van Tilborg, Angela A G; Kompier, Lucie C; Lurkin, Irene

    2012-01-01

    . Moreover, stutter peaks may complicate the analysis. To use microsatellite markers for diagnosis of recurrent bladder cancer, we aimed to select markers without stutter peaks and a constant ratio between alleles, thereby avoiding the need for a control DNA sample. We investigated 49 microsatellite markers...... with tri- and tetranucleotide repeats in regions commonly lost in bladder cancer. Based on analysis of 50 blood DNAs the 12 best performing markers were selected with few stutter peaks and a constant ratio between peaks heights. Per marker upper and lower cut off values for allele ratios were determined...

  9. Measurement-based quantum repeaters

    CERN Document Server

    Zwerger, M; Briegel, H J

    2012-01-01

    We introduce measurement-based quantum repeaters, where small-scale measurement-based quantum processors are used to perform entanglement purification and entanglement swapping in a long-range quantum communication protocol. In the scheme, pre-prepared entangled states stored at intermediate repeater stations are coupled with incoming photons by simple Bell-measurements, without the need of performing additional quantum gates or measurements. We show how to construct the required resource states, and how to minimize their size. We analyze the performance of the scheme under noise and imperfections, with focus on small-scale implementations involving entangled states of few qubits. We find measurement-based purification protocols with significantly improved noise thresholds. Furthermore we show that already resource states of small size suffice to significantly increase the maximal communication distance. We also discuss possible advantages of our scheme for different set-ups.

  10. A Repeating Fast Radio Burst

    CERN Document Server

    Spitler, L G; Hessels, J W T; Bogdanov, S; Brazier, A; Camilo, F; Chatterjee, S; Cordes, J M; Crawford, F; Deneva, J; Ferdman, R D; Freire, P C C; Kaspi, V M; Lazarus, P; Lynch, R; Madsen, E C; McLaughlin, M A; Patel, C; Ransom, S M; Seymour, A; Stairs, I H; Stappers, B W; van Leeuwen, J; Zhu, W W

    2016-01-01

    Fast Radio Bursts are millisecond-duration astronomical radio pulses of unknown physical origin that appear to come from extragalactic distances. Previous follow-up observations have failed to find additional bursts at the same dispersion measures (i.e. integrated column density of free electrons between source and telescope) and sky position as the original detections. The apparent non-repeating nature of the fast radio bursts has led several authors to hypothesise that they originate in cataclysmic astrophysical events. Here we report the detection of ten additional bursts from the direction of FRB121102, using the 305-m Arecibo telescope. These new bursts have dispersion measures and sky positions consistent with the original burst. This unambiguously identifies FRB121102 as repeating and demonstrates that its source survives the energetic events that cause the bursts. Additionally, the bursts from FRB121102 show a wide range of spectral shapes that appear to be predominantly intrinsic to the source and wh...

  11. Repeatability of Harris Corner Detector

    Institute of Scientific and Technical Information of China (English)

    HU Lili

    2003-01-01

    Interest point detectors are commonly employed to reduce the amount of data to be processed. The ideal interest point detector would robustly select those features which are most appropriate or salient for the application and data at hand. This paper shows that interest points are geometrically stable under different transformations.This property makes interest points very successful in the context of image matching. To measure this property quantatively, we introduce a evaluation criterion: repeatability rate.

  12. The Forewarning Effect of Coherence Markers in Persuasive Discourse: Evidence from Persuasion and Processing

    Science.gov (United States)

    Kamalski, Judith; Lentz, Leo; Sanders, Ted; Zwaan, Rolf A.

    2008-01-01

    Several studies showed how coherence markers, like connectives and lexical cue phrases, influence the processing and representation of informative text. Although discourse analysts have repeatedly argued that coherence markers influence the processing of persuasive text as well, there is hardly any empirical evidence for this idea. This article…

  13. Development and mapping of a public reference set of SSR markers in Lolium perenne L.

    NARCIS (Netherlands)

    Bach, J.L.; Muylle, H.; Arens, P.F.P.; Andersen, C.H.; Bach Holm, P.; Ghesquiere, M.; Julier, B.; Lubberstedt, T.; Nielsen, K.K.; Riek, de J.; Roldán-Ruiz, I.; Roulund, N.; Taylor, C.; Vosman, B.J.; Barre, P.

    2005-01-01

    We report on the characterization and mapping of 76 simple sequence repeat (SSR) markers for Lolium perenne. These markers are publicly available or obtained either from genomic libraries enriched for SSR motifs or L. perenne expressed sequence tag (EST) clones. Four L. perenne mapping populations w

  14. Development and transferability of black and red raspberry microsatellite markers from short-read sequences

    Science.gov (United States)

    The advent of next-generation sequencing technologies has been a boon to the cost-effective development of molecular markers, particularly in non-model species. Here, we demonstrate the efficiency of microsatellite or simple sequence repeat (SSR) marker development from short-read sequences using th...

  15. Study on the Origin of Tree Peony Cultivars from Southwest China Based on ISSR Technology%西南牡丹品种起源的ISSR研究

    Institute of Scientific and Technical Information of China (English)

    李宗艳; 秦艳玲; 蒙进芳; 唐岱; 王锦

    2015-01-01

    Objective] Forty-one tree peony samples consisting of 21 from Xinan group, 18 from Zhongyuan group, 1 from Jiangnan group and 1 wild species were used to detect their genetic diversity and verify their phylogenetic relationship and discuss their genetic backgrounds for the new cultivars breeding.[Method]Total genomic DNA was extracted from the fresh leaves of tree peony by a modified CTAB method. A total of 27 primers were selected from 60 ISSR universal primers designed by UBC on the basis of the establishment of an optimal ISSR-PCR reaction system. They were used in the PCR amplification to compare the genetic difference among different cultivars. Comparative analysis on DNA fragments amplified by ISSR-PCR technology was made by the relative genetic software. Genetic parameter such as percentage of polymorphic loci (PPB) was calculated by using POPGENE 32. Cluster analysis (UPGMA) and dendrogram based on Nei’s genetic distance were made to construct the relationship between cultivars and cultivars groups by using the Numerical Taxonomy Multivariate Analysis System (NTSYS-pc) ver. 2.1 statistical package.[Result]A total of 317 bands were obtained by amplifying 41 cultivars from 27 primers, among which 304 bands were polymorphic and percentage of polymorphic bands (PPB) attained to 95.41%. In average, 11.74 bands were produced by each primer. The genetic similarity coefficients among all the tested 41 samples ranged from 0.483 to 0.811, which ‘Daguanfen 4’ and ‘Lijiangzi 5’ had the highest similarity coefficient with 0.811, otherwise ‘Daguanfen 4’and ‘Caihui’ had the lowest coefficient with 0.483. The 41 cultivars were divided into four branches based on UPGMA cluster at the coefficiency of 0.625. The first branch was composed of 19 samples, which included 5 Zhongyuan peony cultivars and 14 Xinan cultivars, the genetic similarity coefficient of ‘Lijiangzi 5’ and ‘Daguanfen 4’ had the closest phylogenetic relationship, ‘Lijiangzi 5’ and

  16. Development of Pineapple Microsatellite Markers and Germplasm Genetic Diversity Analysis

    Directory of Open Access Journals (Sweden)

    Suping Feng

    2013-01-01

    Full Text Available Two methods were used to develop pineapple microsatellite markers. Genomic library-based SSR development: using selectively amplified microsatellite assay, 86 sequences were generated from pineapple genomic library. 91 (96.8% of the 94 Simple Sequence Repeat (SSR loci were dinucleotide repeats (39 AC/GT repeats and 52 GA/TC repeats, accounting for 42.9% and 57.1%, resp., and the other three were mononucleotide repeats. Thirty-six pairs of SSR primers were designed; 24 of them generated clear bands of expected sizes, and 13 of them showed polymorphism. EST-based SSR development: 5659 pineapple EST sequences obtained from NCBI were analyzed; among 1397 nonredundant EST sequences, 843 were found containing 1110 SSR loci (217 of them contained more than one SSR locus. Frequency of SSRs in pineapple EST sequences is 1SSR/3.73 kb, and 44 types were found. Mononucleotide, dinucleotide, and trinucleotide repeats dominate, accounting for 95.6% in total. AG/CT and AGC/GCT were the dominant type of dinucleotide and trinucleotide repeats, accounting for 83.5% and 24.1%, respectively. Thirty pairs of primers were designed for each of randomly selected 30 sequences; 26 of them generated clear and reproducible bands, and 22 of them showed polymorphism. Eighteen pairs of primers obtained by the one or the other of the two methods above that showed polymorphism were selected to carry out germplasm genetic diversity analysis for 48 breeds of pineapple; similarity coefficients of these breeds were between 0.59 and 1.00, and they can be divided into four groups accordingly. Amplification products of five SSR markers were extracted and sequenced, corresponding repeat loci were found and locus mutations are mainly in copy number of repeats and base mutations in the flanking region.

  17. 三个新的PAH基因短串联重复序列多态性及其在苯丙酮尿症产前诊断中的应用%The power of linkage analysis on PAH gene in prenatal gene diagnosis is improved with three additional short tandem repeat markers

    Institute of Scientific and Technical Information of China (English)

    姚凤霞; 郭辉; 韩娟娟; 孟岩; 孙念怙; 黄尚志

    2007-01-01

    目的 提高经典型苯丙酮尿症的产前诊断的成功率.方法 在苯丙氨酸羟化酶(phenylalanine hydroxylase, PAH)基因附近选择了3个新的短串联重复序列(short tandem repeat, STR)位点(PAH26、PAH32和 PAH9),进行扩增长度片段多态性分析,确定它们在中国人群的分布及在诊断中的应用价值.结果 3个新的STR位点的多态信息含量分别为0.518(PAH26)、0.413(PAH32)和0.362(PAH9).这3个位点之间,PAH9与第3内含子中的STR位点(TCTA)n之间存在连锁不平衡,其他位点组合不存在连锁不平衡.联合第3内含子中的STR位点(TCTA)n和2个新的位点,可以对家系中突变基因标记进行95%判断,并成功地用于4例产前诊断中.结论 选择性地应用PAH基因中的3个STR位点组合,可以95%地区分经典型苯丙酮尿症家系中父母的两个基因,从而准确地进行快速产前诊断.

  18. Intra-genomic variation in the ribosomal repeats of nematodes.

    Directory of Open Access Journals (Sweden)

    Holly M Bik

    Full Text Available Ribosomal loci represent a major tool for investigating environmental diversity and community structure via high-throughput marker gene studies of eukaryotes (e.g. 18S rRNA. Since the estimation of species' abundance is a major goal of environmental studies (by counting numbers of sequences, understanding the patterns of rRNA copy number across species will be critical for informing such high-throughput approaches. Such knowledge is critical, given that ribosomal RNA genes exist within multi-copy repeated arrays in a genome. Here we measured the repeat copy number for six nematode species by mapping the sequences from whole genome shotgun libraries against reference sequences for their rRNA repeat. This revealed a 6-fold variation in repeat copy number amongst taxa investigated, with levels of intragenomic variation ranging from 56 to 323 copies of the rRNA array. By applying the same approach to four C. elegans mutation accumulation lines propagated by repeated bottlenecking for an average of ~400 generations, we find on average a 2-fold increase in repeat copy number (rate of increase in rRNA estimated at 0.0285-0.3414 copies per generation, suggesting that rRNA repeat copy number is subject to selection. Within each Caenorhabditis species, the majority of intragenomic variation found across the rRNA repeat was observed within gene regions (18S, 28S, 5.8S, suggesting that such intragenomic variation is not a product of selection for rRNA coding function. We find that the dramatic variation in repeat copy number among these six nematode genomes would limit the use of rRNA in estimates of organismal abundance. In addition, the unique pattern of variation within a single genome was uncorrelated with patterns of divergence between species, reflecting a strong signature of natural selection for rRNA function. A better understanding of the factors that control or affect copy number in these arrays, as well as their rates and patterns of evolution

  19. Markers of erectile dysfunction

    Directory of Open Access Journals (Sweden)

    Kelvin P Davies

    2008-01-01

    Full Text Available With the development and marketing of oral pharmacotherapy that is both noninvasive and successful in treating erectile dysfunction (ED, the quest to identify markers of organic ED lost ground. Indeed, the multi-factorial nature of ED may have led many researchers to conclude that searching for a universal marker of ED was futile. However, the realization that ED is strongly correlated with the overall health of men, and may act as a predictor for the development of cardiovascular disease (CVD and diabetes, has stimulated interest in identifying genes that can distinguish organic ED. In addition, the potential ability to suggest to the patient that ED is reversible (i.e., psychogenic with a simple test would be of significance to both the physician and patient, as well as for reimbursement issues for therapy by insurance companies. Such a marker may also act as a non-subjective measure of the degree of ED and the efficacy of treatment. This review discusses the importance of identifying such markers and recent work identifying potential markers in human patients.

  20. High Resolution Microsatellite Marker Analysis of Some Rice Landraces Using Metaphor Agarose Gel Electrophoresis

    OpenAIRE

    K. Kristamtini; T. Taryono; Panjisakti Basunanda; Rudi Hari Murti

    2016-01-01

    Microsatellite markers or simple sequences repeats are DNA - based molecular