WorldWideScience

Sample records for rapid plant regeneration

  1. Rapid plant regeneration of chrysanthemum ( Chrysanthemum ...

    African Journals Online (AJOL)

    Shoot multiplication of chrysanthemum was achieved from shoot tip explant, using MS media supplemented with different concentrations and combinations of plant growth regulators. Different parameters including shoot initiation percentage, average number of shoots per explant, length of shoots (cm), number of leaves per ...

  2. Rapid plant regeneration of chrysanthemum (Chrysanthemum ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-05-04

    May 4, 2009 ... Shoot multiplication of chrysanthemum was achieved from shoot tip explant, using MS media supplemented with different concentrations and combinations of plant growth regulators. Different parameters including shoot initiation percentage, average number of shoots per explant, length of shoots (cm) ...

  3. Establishment of a Rapid Plant Regeneration System in Physalis angulata L. through Axillary Meristems

    Directory of Open Access Journals (Sweden)

    Owk ANIEL KUMAR

    2015-12-01

    Full Text Available An optimal plant propagation method of Physalis angulata L., a medicinally important herbaceous plant species has been developed using axillary meristem explants. Shoot bud proliferation was initiated from axillary meristem explants cultured on MS medium supplemented with various concentrations of 0.5-2.5mg/L/(BAP/(Zeatin/(KIN. The maximum in vitro response of shooting frequency of explants (88.1% and shoots per explant (42 was achieved with medium containing 1.0mg/L BAP. Multiple shoot culture was established by repeated subculturing of the shoot buds of axillary meristems on shoot multiplication medium. Among the subculture media BAP in combination with 1.5mg/L (IAA+0.25mg/L(GA3 produced maximum shoots per explant (128±0.29 after two weeks of culture. Effective in vitro shoot elongation and rooting was achieved on 1.0mg/L(GA3 and 1.0mg/L(IBA, respectively. Most of the generated shoots were successfully transferred to soil under field conditions. The survival percentage of the transferred plants on soil was found to be 90 per cent.  This protocol can be used for commercial propagation and for future genetic improvement studies.

  4. Direct shoot organogenesis and plant regeneration from ...

    African Journals Online (AJOL)

    Using the whole cotyledonary node as explants, a rapid and efficient regeneration protocol was established for kenaf (Hibiscus cannabinus L.) on Murashige and Skoog (MS) basal medium. The effects of the plant growth regulators: N6-benzyladenine (BA), indole-3-aceticacid (IAA) and the non-ironic surfactant pluronic ...

  5. Establishment of plant regeneration system from anther culture of ...

    African Journals Online (AJOL)

    Frequencies of callus induction and shoot regeneration were 100 and 70.5%, respectively with the whole regeneration procedure completed in 40 days under light. This highly efficient, rapid regeneration system can be applied for both genetic transformation and doubled haploid plant induction. Key words: Tagetes patula, ...

  6. Comparison of Callus induction and plant regeneration

    African Journals Online (AJOL)

    Sang-Hoon Lee

    2012-02-21

    Feb 21, 2012 ... regeneration frequency from transgenic callus, but also useful for molecular breeding of tall fescue through ... Plant regeneration. The plant regeneration culture medium (Lee et al., 2007) was used to regenerate plants from the mature seed-derived calluses. It was ..... bioassays with tobacco tissue cultures.

  7. Screening of a broad range of rice (Oryza sativa L.) germplasm for in vitro rapid plant regeneration and development of an early prediction system

    Science.gov (United States)

    Rice has emerged as a model monocot for studies in agriculture and biotechnology, due to its relatively small genome and ready access to plant material. Tissue culture is one of the tools required for genetic transformation, for some breeding programs, and selection of high frequency regenerator ty...

  8. Establishment of a Rapid Plant Regeneration System in Physalis angulata L. through Axillary Meristems

    Directory of Open Access Journals (Sweden)

    Owk ANIEL KUMAR

    2015-12-01

    Full Text Available An optimal plant propagation method of Physalis angulata L., a medicinally important herbaceous plant species has been developed using axillary meristem explants. Shoot bud proliferation was initiated from axillary meristem explants cultured on MS medium supplemented with various concentrations of 0.5-2.5mg/L/(BAP/(Zeatin/(KIN. The maximum in vitro response of shooting frequency of explants (88.1% and shoots per explant (42 was achieved with medium containing 1.0mg/L BAP. Multiple shoot culture was established by repeated subculturing of the shoot buds of axillary meristems on shoot multiplication medium. Among the subculture media BAP in combination with 1.5mg/L (IAA+0.25mg/L(GA3 produced maximum shoots per explant (128±0.29 after two weeks of culture. Effective in vitro shoot elongation and rooting was achieved on 1.0mg/L(GA3 and 1.0mg/L(IBA, respectively. Most of the generated shoots were successfully transferred to soil under field conditions. The survival percentage of the transferred plants on soil was found to be 90 per cent.  This protocol can be used for commercial propagation and for future genetic improvement studies.

  9. How-To-Do-It: Plant Regeneration.

    Science.gov (United States)

    Pietraface, William J.

    1988-01-01

    Describes a procedure for the growth of tobacco plants in flasks. Demonstrates plant tissue culture manipulation, totipotency, and plant regeneration in approximately 12 weeks. Discusses methods, materials, and expected results. (CW)

  10. High-efficiency regeneration of peanut (Arachis hypogaea L.) plants ...

    African Journals Online (AJOL)

    Jane

    2011-10-03

    Oct 3, 2011 ... Somatic embryogenesis from the axillary meristems of peanut (Arachis hypogaea L.). Plant Biotechnol. Rep. 3(4): 333-340. Srinivasan T, Kumar K, Kirti P (2010). Establishment of efficient and rapid regeneration system for some diploid wild species of Arachis. Plant Cell Tissue Org. Cult. 101(3): 303-309.

  11. Adventitious shoots induction and plant regeneration from ...

    African Journals Online (AJOL)

    A highly efficient regeneration system is a prerequisite step for successful genetic transformation of watermelon cultivars (Citrullus lanatus L.). The objective of this study was to establish efficient in vitro plant regeneration for three watermelon cultivars. To achieve optimal conditions for adventitious shoot induction, the ...

  12. Adventitious shoots induction and plant regeneration from ...

    African Journals Online (AJOL)

    Aghomotsegin

    2015-07-08

    Jul 8, 2015 ... A highly efficient regeneration system is a prerequisite step for successful genetic transformation of watermelon cultivars (Citrullus lanatus L.). The objective of this study was to establish efficient in vitro plant regeneration for three watermelon cultivars. To achieve optimal conditions for adventitious.

  13. Plant regeneration protocol of Andrographis paniculata (Burm. f ...

    African Journals Online (AJOL)

    Gyana Sir

    2013-09-25

    Sep 25, 2013 ... Rapid direct plant regeneration of Andrographis paniculata was achieved from leaf and stem explants on Murashige and Skoog (MS) basal medium supplemented with 1.5 to 3.0 mg/l 6-benzyladenine (BA),. 50 mg/l adenine sulfate (Ads) and 3% (m/v) sucrose. Inclusion of 1.0 mg/l 1-naphthalene acetic acid.

  14. An efficient somatic embryogenesis based plant regeneration from ...

    African Journals Online (AJOL)

    An efficient and rapid somatic embryogenesis based plant regeneration from hypocotyls of three Catharanthus roseus cultivars, Pacifica cherry red (PCR), Heatwave mix color (HWMC), and Mediterranean Rose Red (MRR) was standardized. Hypocotyl-derived primary calluses (HPC) were formed on callus induction ...

  15. In Vitro Regeneration of Endangered Medicinal Plant Heliotropium kotschyi (Ramram).

    Science.gov (United States)

    Sadeq, Manal Ahmed; Pathak, Malabika Roy; Salih, Ahmed Ali; Abido, Mohammed; Abahussain, Asma

    2016-01-01

    Heliotropium kotschyi (Ramram) is an important endangered medicinal plant distributed in the Kingdom of Bahrain. Plant tissue culture technique is applied for ex situ conservation study. Nodal stem segments are cultured in modified MS media supplemented with various combination and concentration of plant growth regulators (PGRs). Plants are regenerated via shoot organogenesis from the nodal meristems. Plants are regenerated in three different steps: initial shoot development, shoot multiplication, and rooting. After 4 weeks of culture, 100 % explants respond to shoot initiation on the medium containing 8.88 μM BAP and 5.71 μM IAA. The highest frequency of shoot regeneration is observed in the same media after second subculture of shoots. The highest rooting frequency is observed in the presence of 2.85 μM IAA. After root development, the plantlets are transferred to pots filled with soil and 60 % of plants survived after 45 days. This plant regeneration protocol is of great value for rapid desert plant propagation program.

  16. Direct shoot organogenesis and plant regeneration from ...

    African Journals Online (AJOL)

    Jane

    2010-12-13

    Dec 13, 2010 ... Kenaf cultivar P3B was conserved in the Laboratory of Bastfibre. Crops of Guangxi University. Seeds were ..... Planta,. 47: 585-588. Srivatanakul M, Park SH, Sanders JR, Salas MG, Smith RH (2000). Multiple shoot regeneration of kenaf (Hibiscus cannabinus L.) from a shoot apex culture system. Plant Cell ...

  17. Plant Regeneration and Genetic Transformation in Eggplant ...

    African Journals Online (AJOL)

    Dr Harmander Gill

    2014-02-05

    Feb 5, 2014 ... http://www.academicjournals.org/AJB. African Journal of Biotechnology. Review. Plant regeneration in eggplant (Solanum melongena L.): A review. M. K. Sidhu1*, A. S. Dhatt1 and G. S. Sidhu2 ... economically important genes for the improvement of eggplant. Key words: Callus, somatic embryogenesis, ...

  18. Somatic embryogenesis and plant regeneration of recalcitrant ...

    African Journals Online (AJOL)

    Many restrictive factors still remain in cotton tissue culture such as long duration, unpredictability and a high degree of genotype dependence. The main objective of this study was to develop a protocol allowing consistent somatic embryogenesis and plant regeneration from five recalcitrant cotton cultivars. Our results ...

  19. Plant grafting: insights into tissue regeneration.

    Science.gov (United States)

    Melnyk, Charles W

    2017-02-01

    For millennia, people have cut and joined different plants together through a process known as grafting. The severed tissues adhere, the cells divide and the vasculature differentiates through a remarkable process of regeneration between two genetically distinct organisms as they become one. Grafting is becoming increasingly important in horticulture where it provides an efficient means for asexual propagation. Grafting also combines desirable roots and shoots to generate chimeras that are more vigorous, more pathogen resistant and more abiotic stress resistant. Thus, it presents an elegant and efficient way to improve plant productivity in vegetables and trees using traditional techniques. Despite this horticultural importance, we are only beginning to understand how plants regenerate tissues at the graft junction. By understanding grafting better, we can shed light on fundamental regeneration pathways and the basis for self/non-self recognition. We can also better understand why many plants efficiently graft whereas others cannot, with the goal of improving grafting so as to broaden the range of grafted plants to create even more desirable chimeras. Here, I review the latest findings describing how plants graft and provide insight into future directions in this emerging field.

  20. Development of an efficient plant regeneration protocol for sweet ...

    African Journals Online (AJOL)

    UKZN

    2012-10-18

    Oct 18, 2012 ... reproducible plant transformation protocol (Liu and. Cantliffe, 1984). The ability to regenerate a whole plant from isolated plant cells or tissues which have been genetically transformed underpins most plant transformation systems. Therefore, tissue culture and plant regeneration are integral parts of most ...

  1. Establishment of an efficient and rapid method of multiple shoot regeneration and a comparative phenolics profile in in vitro and greenhouse-grown plants of psophocarpus tetragonolobus (L.) DC

    Science.gov (United States)

    Singh, Vinayak; Chauhan, Namita Singh; Singh, Mohit; Idris, Asif; Madanala, Raju; Pande, Veena; Mohanty, Chandra Sekhar

    2014-01-01

    An in vitro method of multiple shoot induction and plant regeneration in Psophocarpus tetragonolobus (L.) DC was developed. Cotyledons, hypocotyls, epicotyls, internodal and young seedling leaves were used as explants. MS media supplemented with various concentrations of either thidiazuron (TDZ) or N6-benzylaminopurine (BAP) along with NAA or IAA combinations were used to determine their influence on multiple shoot induction. MS media supplemented with TDZ induced direct shoot regeneration when epicotyls and internodal segments were used as explants. TDZ at 3 mg L−1 induced highest rate (89.2 ± 3.28%) of regeneration with (13.4 ± 2.04) shoots per explant. MS media supplemented with BAP in combination with NAA or IAA induced callus mediated regeneration when cotyledons and hypocotyls were used as explants. BAP (2.5 mg L−1) and IAA (0.2 mg L−1) induced highest rate (100 ± 2.66%) of regeneration with (23.2 ± 2.66) shoots per explant. Mature plants produced from regenerated shoots were transferred successfully to the greenhouse. In a comparative study, the phenolics contents of various parts of greenhouse-grown plants with that of in vitro-raised plants showed significant variations. PMID:25482808

  2. Development of an efficient plant regeneration protocol for sweet ...

    African Journals Online (AJOL)

    An efficient and reproducible plant regeneration protocol for the South African sweet potato (Ipomoea batatas Lam.) cultivar Blesbok was developed in this study. The effect of different hormone combinations and type of explant on shoot regeneration was evaluated in order to optimize the regeneration protocol. Explants in ...

  3. High-efficiency regeneration of peanut ( Arachis hypogaea L.) plants ...

    African Journals Online (AJOL)

    A high-efficiency regeneration system for peanut plants was established. The regeneration frequency of leaf discs reached 40.9% on Murashige and Skoog medium supplemented with 0.5 mg l-1 naphthylacetic acid and 0.5 mg l-1 thidiazuron. The regenerated shoots elongated, developed roots and produced seeds.

  4. Somatic embryogenesis and plant regeneration from leaf explants of ...

    African Journals Online (AJOL)

    An attempt was made to study the somatic embryogenesis and plant regeneration from the in vitro leaf explants of Rumex vesicarius L. a renowned medicinal plant, which belongs to polygonaceae family. Effective in vitro regeneration of R. vesicarius was achieved via young leaf derived somatic embryo cultures.

  5. Prolific plant regeneration through organogenesis from scalps of ...

    African Journals Online (AJOL)

    Prolific plant regeneration through organogenesis from scalps of Musa sp cv. Tanduk. SMA Elhory, MA Aziz, AA Rashid, AG Yunus. Abstract. A prolific plant regeneration system using scalps derived from shoot tips of Musa spp. cv. Tanduk was developed. Highly proliferating scalps, produced after four monthly subcultures ...

  6. Genotypic variability in callus induction and plant regeneration ...

    African Journals Online (AJOL)

    Depending on different genotypes, the best plant regeneration was obtained on LS (Linsmaier and Skoog, 1965) based medium supplemented with 2 mgl-1 BAP + 1.5 mg l-1 2,4-D. Large variabilities in callus growth and plant regeneration potential were revealed among the cultivars tested. Cultivar HA-8 formed a high ...

  7. Plant regeneration in wheat mature embryo culture | Aydin | African ...

    African Journals Online (AJOL)

    Success in genetic engineering of cereals depends on the callus formation and efficient plant regeneration system. Callus formation and plant regeneration of wheat mature embryos were assessed by using 12 different methods with 4 genotypes. Genotype significantly affected the formation of callus, embryogenic callus ...

  8. Plant regeneration and genetic transformation in Jatropha

    KAUST Repository

    Sujatha, M.

    2012-07-01

    Jatropha curcas, a non-edible oil bearing species with multiple uses, and considerable economic potential is emerging as a potential biofuel plant. The limited knowledge of this species, low and inconsistent yields, the narrow genetic variability, and vulnerability to insects and diseases are major constraints in successful cultivation of Jatropha as a biofuel crop. Hence, genetic improvement of Jatropha is essential by conventional and modern biotechnological tools to use as a viable alternative source of bio-diesel. Realising its potential as a bio-energy crop, in vitro regeneration methods have been established to meet the demand of large scale supply of superior clones, and also as a prelude for genetic improvement of the species through transgenic approaches. In this chapter, an overview of in vitro tissue culture and genetic transformation of Jatropha is discussed. © 2013 Springer Science+Business Media Dordrecht. All rights are reserved.

  9. Optimization of a plant regeneration protocol for broccoli

    African Journals Online (AJOL)

    user

    2011-05-16

    May 16, 2011 ... African Journal of Biotechnology Vol. 10(20), pp. .... these modifications resulted in a notable improvement in the regeneration rate of the .... Plant Cell Rep. 11: 334-338. Birch RG (1997). Plant transformation: problems and strategies for practical application. Annu. Rev. Plant Physiol. Plant Mol. Biol. 48:.

  10. Factors affecting in vitro plant regeneration from cotyledonary node ...

    African Journals Online (AJOL)

    Factors affecting in vitro plant regeneration from cotyledonary node explant of Senna sophera (L.) Roxb. – A highly medicinal legume. ... Keywords: Senna sophera, fabaceae, cotyledonary node, in vitro shoot regeneration, rooting, acclimatization. African Journal of Biotechnology, Vol. 13(3), pp. 413-422, 15 January, 2014 ...

  11. Prolific plant regeneration through organogenesis from scalps of ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-11-16

    Nov 16, 2009 ... Full Length Research Paper. Prolific plant regeneration through organogenesis from scalps of Musa sp ... include the panama disease or Fusarium wilt (Fusarium oxysporum f. sp. Cubense and the black sigatoka (Myco- ... This paper describes prolific shoot regeneration from scalps of Musa sp. cv. Tanduk.

  12. Competence and regulatory interactions during regeneration in plants

    Science.gov (United States)

    Pulianmackal, Ajai J.; Kareem, Abdul V. K.; Durgaprasad, Kavya; Trivedi, Zankhana B.; Prasad, Kalika

    2014-01-01

    The ability to regenerate is widely exploited by multitudes of organisms ranging from unicellular bacteria to multicellular plants for their propagation and repair. But the levels of competence for regeneration vary from species to species. While variety of living cells of a plant display regeneration ability, only a few set of cells maintain their stemness in mammals. This highly pliable nature of plant cells in-terms of regeneration can be attributed to their high developmental plasticity. De novo organ initiation can be relatively easily achieved in plants by proper hormonal regulations. Elevated levels of plant hormone auxin induces the formation of proliferating mass of pluripotent cells called callus, which predominantly express lateral root meristem markers and hence is having an identity similar to lateral root primordia. Organ formation can be induced from the callus by modulating the ratio of hormones. An alternative for de novo organogenesis is by the forced expression of plant specific transcription factors. The mechanisms by which plant cells attain competence for regeneration on hormonal treatment or forced expression remain largely elusive. Recent studies have provided some insight into how the epigenetic modifications in plants affect this competence. In this review we discuss the present understanding of regenerative biology in plants and scrutinize the future prospectives of this topic. While discussing about the regeneration in the sporophyte of angiosperms which is well studied, here we outline the regenerative biology of the gametophytic phase and discuss about various strategies of regeneration that have evolved in the domain of life so that a common consensus on the entire process of regeneration can be made. PMID:24782880

  13. Plant regeneration from callus cultures of Vitex trifolia (Lamiales: Lamiaceae): a potential medicinal plant.

    Science.gov (United States)

    Samantaray, Sanghamitra; Bishoyi, Ashok Kumar; Maiti, Satyabrata

    2013-09-01

    Vitex trifolia is a shrub species with popular use as a medicinal plant, for which leaves, roots and flowers have been reported to heal different distresses. The increasing exploitation of these plants has endangered its conservation, and has importantly justified the use of biotechnological tools for their propagation. Our aim was to present an efficient protocol for plant regeneration through organogenesis; and simultaneously, to analyze the genetic homogeneity of the established clonal lines by Randomly Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeat (ISSR) markers. Plantlet regeneration was achieved in callus cultures derived from stem, leaf and petiole explants of V. trifolia on a differently supplemented Murashige & Skoog medium, and incubated at 25 +/-2 degrees C under a light intensity of 61 micromol/m2s from cool white fluorescent lamps and a 16 h photoperiod. The rate of shoot bud regeneration was positively correlated with the concentration of hormones in the nutrient media. Shoot buds regenerated more rapidly from stem and petiole explants as compared to leaf explants on medium containing 11.10 microM BAP in combination with 0.54 microMNAA. Addition of 135.74-271.50 microM adenine sulphate (Ads) and 0.72-1.44 microM gibberellic acid (GA3) to the culture medium increased the growth of shoot buds. The highest rate of shoot bud regeneration responses was obtained in stem explants using 11.10 microM BAP in combination with 0.54 microM NAA, 271.50 microM Ads and 1.44 microM GA3. In vitro rooting of the differentiated shoots was achieved in media containing 1.23 microM indole butyric acid (IBA) with 2% (w/v) sucrose. Regenerated plantlets were successfully established in soil with 86% survival under field condition. Randomly Amplified Polymorphic DNA and Inter Simple Sequence Repeat markers analyses have confirmed the genetic uniformity of the regenerated plantlets derived from the second up to fifth subcultures. This protocol may help in

  14. In vitro Plant Regeneration of Cyphomandra betacea through Nodal ...

    African Journals Online (AJOL)

    In vitro Plant Regeneration of Cyphomandra betacea through Nodal Culture. Bancy Waweru, Rodrigue Ishimwe, Joëlle Kajuga, Boniface Kagiraneza, Peter Yao Kanze Sallah, Vedaste Ahishakiye, Safia Kalisa, Theodore Asiimwe, Jane Kahia, Daphrose Gahakwa ...

  15. Development of an efficient plant regeneration protocol for sweet ...

    African Journals Online (AJOL)

    UKZN

    2012-10-18

    Oct 18, 2012 ... Key words: Tissue culture, regeneration, sweet potato, genetic transformation. ... documented using different explants including anthers. (Tsay and .... tuberous roots and morphological observations, including plant stature ...

  16. In Vitro Plant Regeneration from Commercial Cultivars of Soybean

    Directory of Open Access Journals (Sweden)

    Ghulam Raza

    2017-01-01

    Full Text Available Soybean, a major legume crop, is the source of vegetable oil and protein. There is a need for transgenic approaches to breeding superior soybean varieties to meet future climate challenges. Efficient plant regeneration is a prerequisite for successful application of genetic transformation technology. Soybean cultivars are classified into different maturity groups based on photoperiod requirements. In this study, nine soybean varieties belonging to different maturity group were regenerated successfully from three different explants: half split hypocotyl, complete hypocotyl, and cotyledonary node. All the genotypes and explant types responded by producing adventitious shoots. Shoot induction potential ranged within 60–87%, 50–100%, and 75–100%, and regeneration rate ranged within 4.2–10, 2.7–4.2, and 2.6–10.5 shoots per explant using half split hypocotyl, complete hypocotyl, and cotyledonary explants, respectively, among all the tested genotypes. Bunya variety showed the best regeneration response using half split and complete hypocotyl explants and the PNR791 with cotyledonary node. The regenerated shoots were successfully rooted and acclimatized to glasshouse conditions. This study shows that commercial varieties of soybean are amenable to shoot regeneration with high regeneration frequencies and could be exploited for genetic transformation. Further, our results show no correlation between shoots regeneration capacity with the maturity grouping of the soybean cultivars tested.

  17. Transformation and regeneration of the holoparasitic plant Phelipanche aegyptiaca

    Directory of Open Access Journals (Sweden)

    Fernández-Aparicio Mónica

    2011-11-01

    Full Text Available Abstract Background Transformation and subsequent regeneration of holoparasitic plants has never been reported, in part due to challenges in developing transformation protocols, but also because regeneration of obligate parasites is difficult since their survival depends completely on successful haustorium penetration of a host and the formation of vascular connections. The recent completion of a massive transcriptome sequencing project (the Parasitic Plant Genome Project will fuel the use of genomic tools for studies on parasitic plants. A reliable system for holoparasite transformation is needed to realize the full value of this resource for reverse genetics and functional genomics studies. Results Here we demonstrate that transformation of Phelipanche aegyptiaca is achieved by infection of 3 month-old in vitro grown P. aegyptiaca calli with Agrobacterium rhizogenes harboring the yellow fluorescent protein (YFP. Four months later, YFP-positive regenerated calli were inoculated onto tomato plants growing in a minirhizotron system. Eight days after inoculation, transgenic parasite tissue formed lateral haustoria that penetrated the host and could be visualized under UV illumination through intact host root tissue. YFP-positive shoot buds were observed one month after inoculation. Conclusions This work constitutes a breakthrough in holoparasitic plant research methods. The method described here is a robust system for transformation and regeneration of a holoparasitic plant and will facilitate research on unique parasitic plant capabilities such as host plant recognition, haustorial formation, penetration and vascular connection.

  18. Plant regeneration via somatic embryogenesis from root explants of ...

    African Journals Online (AJOL)

    A system for induction of callus and plant regeneration via somatic embryogenesis from root explants of Hevea brasiliensis Muell. Arg. clone Reyan 87-6-62 was evaluated. The influence of plant growth regulators (PGRs) including 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (6-BA) and kinetin (KT) on ...

  19. Plant regeneration of Lotononis bainesii Baker (Fabaceae) through ...

    African Journals Online (AJOL)

    Lotononis bainesii Baker is a promising perennial forage legume for subtropical regions. The development of tissue culture methods for in vitro plant regeneration is useful, for example, for the propagation of selected plants and germplasm conservation. In addition, it could also facilitate crop improvement methods. For this ...

  20. Plant Regeneration and Stimulation of in vitro flowering in Eruca ...

    African Journals Online (AJOL)

    hcl

    2012-04-17

    Apr 17, 2012 ... Plant 38: 513-518. Yaniv Z, Schafferman D, Amar Z (1998). Tradition, Uses, and. Biodiversity of Rocket (Eruca sativa) in Israel. Econ. Bot. 52: 394-. 400. Zhang T, Cao ZY, Wang XY (2005). Induction of somatic embryogenesis and plant regeneration from cotyledon and hypocotyl explants of Eruca sativa Mill.

  1. Efficient callus induction and indirect plant regeneration from various ...

    African Journals Online (AJOL)

    The Jatropha curcas is considered as an important energy plant due to the fact that its seed contains high oil content. Nowadays focus is being placed on J. curcas callus induction and plant regeneration. In this study, explants epicotyl, hypocotyl, petiole and cotyledon of 8-day-old seedlings of J. curcas were utilized for ...

  2. Plant regeneration system from cotyledons-derived calluses cultures ...

    African Journals Online (AJOL)

    Administrator

    2011-09-26

    Sep 26, 2011 ... 3 Key Laboratory of Protection and Development Utilization of Tropical Crop Germplasm Resources (Hainan University),. Ministry of Education ..... Regeneration of isolated mesophyll and cell suspension protoplasts to plants in Stylosanthes guianensis. A tropical forage legume. Plant Cell Rep. 3: 174-177.

  3. Effect of plant growth regulators on regeneration of the endangered ...

    African Journals Online (AJOL)

    The use of in vitro techniques for conserving plant biodiversity and protecting rare and endangered multipurpose plant species is considered as one of the most important ex-situ conservation policies. Development of an efficient in vitro regeneration protocol of Calligonum comosum is important and that has achieved to ...

  4. Efficient plant regeneration from leaf explants of Solanum americanum

    African Journals Online (AJOL)

    A very efficient system for direct plant regeneration from in vitro–derived leaf explants of Solanum americanum was developed. S. americanum is a tropical plant with important medical properties. The in vitro procedure that was established consists of (i) induction of shoots from leaf tissue, (ii) elongation of shoots, and (iii) ...

  5. Full Length Research Paper Plant regeneration of Michelia ...

    African Journals Online (AJOL)

    Michelia champaca L. is a woody ornamental tree species which has high commercial value to be used as a basic material for perfume, cosmetic, and medicine. The development of an efficient plant regeneration system for M. champaca is essential for the production of Champaca planting material and precondition for ...

  6. Factors influencing callus induction and plant regeneration of ...

    African Journals Online (AJOL)

    ajl yemi

    2012-01-12

    Jan 12, 2012 ... transgenic perennial ryegrass (Lolium perenne L.) obtained by. Agrobacterium tumefaciens-mediated transformation of the vacuolar. Na+/H+ antiporter gene. Plant Sci. 169: 65-73. Zhao J, Zhou C, Yang HY (1999). In vitro development of early proembryos and plant regeneration via microculture in Oryza ...

  7. Plant regeneration studies of Jatropha curcas using induced ...

    African Journals Online (AJOL)

    A plant regeneration system for Jatropha curcas, a biofuel- producing plant, was established from the induced embryogenic callus. Cotyledon explants cultured on single auxin media were able to form callus. However, only the callus induced from cotyledon explants using Murashige and Skoog (MS) medium containing 0.8 ...

  8. Adventitious shoots induction and plant regeneration from ...

    African Journals Online (AJOL)

    Aghomotsegin

    2015-07-08

    Jul 8, 2015 ... cultivars were placed on MB5 media supplemented with different concentrations and combinations of growth regulators (1.0 ... Key words: Watermelon (Citrullus lanatus), cotyledon, growth regulator, kanamycin, regeneration. ..... expression of Saccharomyces cerevisiae HAL1 gene increases salt tolerance ...

  9. Plant regeneration through indirect organogenesis of chestnut ...

    African Journals Online (AJOL)

    Mehrcedeh

    2013-12-18

    Dec 18, 2013 ... The highest shoot multiplication (66.9%) was observed on. MS medium with 0.2 mgLˉ¹ TDZ. Regenerated shoots were rooted in vitro on MS containing 1.5 mgLˉ¹. IBA. Also, plantlets with well developed root and shoot systems were acclimatized inside the green house and 80% of the plantlets survived ...

  10. Vegetative Regeneration Capacities of Five Ornamental Plant Invaders After Shredding

    Science.gov (United States)

    Monty, Arnaud; Eugène, Marie; Mahy, Grégory

    2015-02-01

    Vegetation management often involves shredding to dispose of cut plant material or to destroy the vegetation itself. In the case of invasive plants, this can represent an environmental risk if the shredded material exhibits vegetative regeneration capacities. We tested the effect of shredding on aboveground and below-ground vegetative material of five ornamental widespread invaders in Western Europe that are likely to be managed by cutting and shredding techniques: Buddleja davidii (butterfly bush, Scrophulariaceae), Fallopia japonica (Japanese knotweed, Polygonaceae), Spiraea × billardii Hérincq (Billard's bridewort, Rosaceae), Solidago gigantea (giant goldenrod, Asteraceae), and Rhus typhina L. (staghorn sumac, Anacardiaceae). We looked at signs of vegetative regeneration and biomass production, and analyzed the data with respect to the season of plant cutting (spring vs summer), the type of plant material (aboveground vs below-ground), and the shredding treatment (shredded vs control). All species were capable of vegetative regeneration, especially the below-ground material. We found differences among species, but the regeneration potential was generally still present after shredding despite a reduction of growth rates. Although it should not be excluded in all cases (e.g., destruction of giant goldenrod and staghorn sumac aboveground material), the use of a shredder to destroy woody alien plant material cannot be considered as a general management option without significant environmental risk.

  11. High frequency plant regeneration from shoot tip explants of ...

    African Journals Online (AJOL)

    A high frequency and rapid regeneration protocol was developed from shoot tip explants of Citrullus colocynthis on Murashige and Skoog (MS) medium supplemented with N6-benzylamino-purine (BAP, 0.5 mg/l) and α-naphthalene acetic acid (NAA, 0.5 mg/l). Highest number of shoots (23.0 ± 0.567) was obtained on MS ...

  12. In vitro callus induction and plant regeneration from mature seed ...

    African Journals Online (AJOL)

    The method for callus induction, adventitious shoot induction and plant regeneration derived from mature embryos of the seed and young shoots in Dendrocalamus farinosus was examined. For young shoots, the lowest callus induction frequency (21.0 to 29.7%) was obtained and the induced callus became brown and ...

  13. Establishment of a plant regeneration system from callus of ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-11-19

    Nov 19, 2008 ... Establishment of a plant regeneration system from callus of Dendrobium cv. Serdang Beauty. Khosravi, A. R.1, Kadir, M. A.1, Kazemin, S. B.2, Zaman F.Q3 and De Silva, A. E.3 ... An in vitro propagation protocol was established for the Dendrobium Serdang Beauty orchid. ..... (Bar=Standard deviation, n=10).

  14. In vitro plant regeneration from Narbon Vetch ( Vicia narbonensis L ...

    African Journals Online (AJOL)

    Narbon vetch (Vicia narbonensis L.) is an agriculturally important forage plant that widely grows in an area extending from Central Europe to various parts of Asia. The study reports axillary shoot regeneration from cotyledonary node explants obtained from, in vitro raised seeds of 4 - 5 and 14 - 15 days old seedlings on MS ...

  15. Plant regeneration system from cotyledons-derived calluses cultures ...

    African Journals Online (AJOL)

    The objective of this study was to successfully establish plant regeneration system with cotyledons of Stylosanthes guianensis Sw. cv. 'Reyan 2' as explants. In this study, the following results were obtained; (1) the highest rates of callus induction on medium MS with 3.0 mg L-1 2, 4-D with cotyledons as explants were 74%.

  16. Plants regeneration from African cowpea variety ( Vigna unguiculata ...

    African Journals Online (AJOL)

    Vigna unguiculata (L.) Walp. plant was efficiently regenerated from cotyledonary node explants. The shoots multiplication rate was influenced by the presence or the absence of cotyledons. Explants with two entire cotyledons from 5-6-d-old seedling produced the greater number of shoots (8.30) after two weeks on B5 ...

  17. Induction of somatic embryogenesis and plant regeneration in the ...

    African Journals Online (AJOL)

    An in vitro culture system for the large-scale propagation of Phragmites communis Trin. (reed) was established by optimizing culture conditions for callus induction and differentiation together with plant propagation using regenerated plantlets. Callus was induced from stem segments with callus induction medium containing ...

  18. Plant regeneration of Lotononis bainesii Baker (Fabaceae) through ...

    African Journals Online (AJOL)

    María Laura Vidoz

    2012-05-22

    May 22, 2012 ... protocol for different genotypes within a cultivar and from different explants of L. bainesii. Plant regeneration was ..... Physiol. Planta. 94: 268-276. Ozias-Akins P, Gill R (2001). Progress in the development of tissue culture and transformation methods applicable to the production of transgenic peanut. Peanut ...

  19. Establishment of a plant regeneration system from callus of ...

    African Journals Online (AJOL)

    Establishment of a plant regeneration system from callus of Dendrobium cv. Serdang Beauty. AR Khosravi, MA Kadir, SB Kazemin, FQ Zaman, AE De Silva. Abstract. An in vitro propagation protocol was established for the Dendrobium Serdang Beauty orchid. The propagation protocol utilized calli tissues that were ...

  20. Direct and indirect plant regeneration from various explants of ...

    African Journals Online (AJOL)

    Populus species are important resource for certain branches of industry and have special roles for scientific study on biological and agricultural systems. Plant regeneration via direct and indirect organogenesis of four Populus deltoides Bartram ex Marsh. ssp. deltoides × Populus deltoides Bartram ex Marsh. ssp. deltoides ...

  1. Direct and indirect plant regeneration from various explants of ...

    African Journals Online (AJOL)

    user

    2011-04-18

    Apr 18, 2011 ... 1700s from North America. In Turkey, it is an exotic species of poplar and was introduced in the late 1960s from the USA. ..... Balkan H, Tanrıyar H, Calikoglu M, Ogras T, Ozden O, Tulukcu M,. Tank T (1998). Genotype differencies in direct plant regeneration from stem explants of Populus tremula in Turkey.

  2. Establishment of in vitro plant regeneration system for ...

    African Journals Online (AJOL)

    An efficient protocol for plant regeneration of Chimonanthus praecox (L.) Link, was developed using leaves from seedlings of seeds. Shoots induction were influenced by cytokinins and nodal positions. The results show that the highest callus induction frequency was obtained on MS medium supplemented with 2.0 mg l−1 ...

  3. Plant regeneration via somatic embryogenesis from root explants of ...

    African Journals Online (AJOL)

    hope&shola

    2010-08-30

    benzylaminopurine (6-BA) and kinetin (KT) on callus induction of root ... The pH of the medium was adjusted to 5.8 and all media were .... embryos; g, successful soil transfer of regenerated plants. Scale bars represent 0.5 cm (b, e), 1 cm ...

  4. Cryopreservation and plant regeneration of anther callus in Hevea ...

    African Journals Online (AJOL)

    Callus induced from anther of Hevea brasiliensis was successfully cryopreserved in liquid nitrogen (LN) by vitrification method and subsequently regenerated into plants. The effects of different preculture time, loading and dehydration duration on callus viability after cryopreservation were evaluated. The effective ...

  5. Effect of genotype on callus induction and plant regeneration from ...

    African Journals Online (AJOL)

    Nine sugarcane genotypes (CP59-73, CP63-588, CP80-314, SP71-1081, F160, L62-96, CP70-321, CP57- 614 and Clone III) were evaluated for their callus induction capacity, embryogenic callus production and plant regeneration ability. Leaf cylinders were used as explants using Murashige and Skoog (MS) based ...

  6. In vitro regeneration of a common medicinal plant, Ocimum sanctum ...

    African Journals Online (AJOL)

    In vitro regeneration of a common medicinal plant, Ocimum sanctum L. for mass propagation. Mohammad Abu Hena Mostofa Jamal, Imdadul Hoque Sharif, Md. Mostofa Shakil, A.N.M. Rubaiyath-Bin Rahman, Nilufa Akhter Banu, Md. Rezuanul Islam, Md. Nazmuzzaman ...

  7. Optimized system for plant regeneration of watermelon (Citrullus ...

    African Journals Online (AJOL)

    Jane

    2011-08-29

    Aug 29, 2011 ... The objective of this study was to establish an efficient and reproducible in vitro plant regeneration for. Citrullus lanatus cv. Zaojia. .... To determine the regional effects of cotyledon for shoot formation, five explants were tested in this ... micro- and macronutrients in culture tubes. After 3 weeks on rooting ...

  8. Somatic embryogenesis and plant regeneration of recalcitrant cottons

    African Journals Online (AJOL)

    STORAGESEVER

    2009-02-04

    Feb 4, 2009 ... regeneration through somatic embryogenesis for cotton cultivars with Fusarium and/or Verticillium wilts resistance. MATERIALS AND METHODS. Plant materials. Five upland cotton (G. hirsutum) cultivars (Shann724, Zhong6331,. CCRI18, Liaomian12 and Jinmian14), provided by Cotton Genetics.

  9. High frequency plant regeneration from shoot tip explants of ...

    African Journals Online (AJOL)

    USER

    2010-08-02

    Aug 2, 2010 ... Sobhakumari VP, Lalithakumari D (2003). Direct plant regeneration from shoot tip cultures of Capsicum annuum L. cv. PLR-1. Phytomorphology, 53(3&4): 235-242. Suchitra B, Mehar Z, Susil K (1999). In vitro multiplication of Centella asiatica. Curr. Sci. 76: 147-158. Sunandakumari C, MaSurtin KP, Chithra ...

  10. Optimization of callus induction and plant regeneration from ...

    African Journals Online (AJOL)

    USER

    2010-04-19

    Apr 19, 2010 ... on 1/2 MS medium and then transplanted to pots, where they exhibited morphologically normal growth. Key words: Embryogenic callus, optimal media, orthogonal test, plant regeneration, range analysis, Sorghum bicolor Moench. ..... fermentations by Saccharomyces cerevisiae. World J. Microbiol.

  11. Factors affecting in vitro plant regeneration from cotyledonary node ...

    African Journals Online (AJOL)

    SHAHINA

    2014-01-15

    Jan 15, 2014 ... in vitro regeneration protocol, where different factors were optimized for maximum multiplication and propagation. The age of the explant, plant growth regulators, basal medium, pH of the medium and sucrose concentrations markedly influenced in vitro propagation of S. sophera. Among 14, 21 and 28.

  12. Plant regeneration from leaf-derived callus in Plectranthus barbatus ...

    African Journals Online (AJOL)

    RAMA

    2013-05-01

    May 1, 2013 ... shoot multiplication was achieved at the 6th subculture and more than 2000 shoots were produced from callus clump. Roots were induced from shoots of in vitro grown plantlets on basal half strength MS medium and high rooting frequencies were obtained. Regenerated plants were easily acclimatized in.

  13. Efficient regeneration of plants from shoot tip explants of ...

    African Journals Online (AJOL)

    Dendrobium densiflorum Lindl. is one of the horticulturally important orchids of Nepal due to its beautiful yellowish flower and medicinal properties. The present study was carried out for plant regeneration from shoot tip explants of D. densiflorum by tissue culture technique. The shoot tip explants of this species, obtained ...

  14. Regeneration of plants from Fraxinus americana hypocotyls and cotyledons

    Science.gov (United States)

    Kaitlin J. Palla; Paula M. Pijut

    2011-01-01

    A plant regeneration protocol was developed for white ash (Fraxinus americana L.). Hypocotyls and cotyledons excised from embryos were cultured on Murashige and Skoog (MS)mediumsupplementedwith 6-benzylaminopurine (BA) plus thidiazuron (TDZ), and compared for organogenic potential. Sixty-six percent of hypocotyl segments and 10.4% of cotyledon...

  15. Establishment of a high-efficiency plant regeneration and ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-10-05

    Oct 5, 2009 ... differentiated into plants, in spite of high CIP. Maize inbred line 18-599(red) as a ... bombardment, after selection and differentiation culture, partial bombarded calli were regenerated into green plantlets and further .... described in the manufacturer's instructions, with 1100 psi helium pressure and a target ...

  16. High frequency plant regeneration from desiccated calli of indica rice

    African Journals Online (AJOL)

    An efficient and reproducible protocol is required to achieve high frequency transformation from transformed calli. We report here high frequency plant regeneration from mature seed derived embryogenic calli of two recalcitrant indica rice cultivars HKR-46 and HKR-126 after partial desiccation treatment. Embryogenic and ...

  17. Studies on plant regeneration and transformation efficiency of ...

    African Journals Online (AJOL)

    We have standardized the tissue culture media for the regeneration and transformation with the vector LBA 4404 (pCAMBIA 2301), so that in future, this system may be exploited for the expression of antibody fragment (single chain variable fragment) in plants (plantibody). The transformed green shoots tested positive for ...

  18. Genetic variation in colchicine-treated regenerated plants of ...

    Indian Academy of Sciences (India)

    Annual Meetings · Mid Year Meetings · Discussion Meetings · Public Lectures · Lecture Workshops · Refresher Courses · Symposia. Home; Journals; Journal of Genetics; Volume 88; Issue 3. Genetic variation in colchicine-treated regenerated plants of Eucalyptus globulus Labill. X. Z. Liu X. Lin X. Y. Mo T. Long H. Y. Zhang.

  19. In vitro plant regeneration from embryogenic cell suspension culture ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-05-02

    May 2, 2008 ... In vitro plant regeneration was achieved from embryogenic cell suspension culture of Astragalus chrysochlorus. When 30-day-old aseptically ... previous study, cytotoxic activities of stem and root ex-. *Corresponding author. E-mail: ... For callus induction, 30-day-old mesocotyl parts of seedlings were used.

  20. Plant regeneration from callus culture of vetiver (Vetiveria zizanioides Nash

    Directory of Open Access Journals (Sweden)

    Somporn Prasertsongskun

    2003-09-01

    Full Text Available The present research aimed to establish cell suspension culture of vetiver (Vetiveria zizanioides Nash from Surat Thani germplasm source and efficient plant regeneration from callus derived from such cultures. Cell suspension cultures were established from calli derived from inflorescence of vetiver. Optimum cell proliferation occurred in liquid N6 medium supplemented with 10 μM 2,4-dichlorophenoxyacetic acid (2,4-D and 10 mM proline. The cell suspension formed the highest small colonies when plated on solid MS medium containing 0.45 μM 2,4-D. After subsequent transfer to regeneration medium (MS free medium 65% of plantlets were obtained.

  1. Somatic embryogenesis and plant regeneration of Capsicum baccatum L.

    Directory of Open Access Journals (Sweden)

    Peddaboina Venkataiah

    2016-06-01

    Full Text Available A plant regeneration protocol via somatic embryogenesis was achieved in cotyledon and leaf explants of Capsicum baccatum, when cultured on MS medium supplemented with various concentrations of 2,4-dichlorophenoxy acetic acid (2,4-D, 0.5–5.0 mg l−1 in combination with Kinetin (Kn, 0.5 mg l−1 and 3% sucrose. Various stages were observed during the development of somatic embryos, including globular, heart, and torpedo-stages. Torpedo stage embryos were separated from the explants and subcultured on medium supplemented with various concentrations of different plant growth regulators for maturation. Maximum percentage (55% of somatic embryo germination and plantlet formation was found at 1.0 mg l−1 BA. Finally, about 68% of plantlets were successfully established under field conditions. The regenerated plants were morphologically normal, fertile and able to set viable seeds.

  2. Tissue culture and regeneration of an antimalarial plant, Artemisia sieberi Besser

    Directory of Open Access Journals (Sweden)

    A. Sharafi

    2014-10-01

    Full Text Available WHO recommends artemisinin-based combination therapies (ACTs as the most effective choice to treat malaria. For developing transgenic plants with high accumulation of artemisinin (by introducing genes encoding enzymes which regulate the biosynthetic pathway of artemisinin, an efficient protocol for tissue culture and plant regeneration is necessary. In the present study, leaf explants of Artemisia sieberi were cultivated in Murashige & Skoog based medium supplemented by combination of different plant growth regulators including 6-benzyl-aminopurine (BA, α-naphthalene-acetic acid (NAA, indole-3-acetic acid (IAA, picloram (Pic and 2,4-dichlorophenoxyacetic acid (2,4-D. The highest frequency of shoot induction was obtained on MS medium supplemented with 2 mg/L BA plus 0.05 mg/L NAA (95% regeneration and MS medium supplemented with 2 mg/L BA plus 0.5 mg/L IAA (85% regeneration. Rooting was obtained on MS medium supplemented with 0.05 mg/L NAA. The present study has revealed a simple, reliable, rapid and high efficient regeneration system for A. sieberi Besser as a source of artemisinin in short period via adventitious shoot induction procedure.

  3. PLANT REGENERATION THROUGH TISSUE CULTURE OF PEAR ...

    African Journals Online (AJOL)

    AISA

    Pennisetum Glaucum (L) R.) K.TIECOURA 1, L. LEDOUX.2 AND M. DINANT.2. 1 Laboratoire de Génétique et amélioration des plantes, UFR de Biosciences, Université de Cocody,. B.P.582 Abidjan 22, Côte d'Ivoire. 2 Laboratoire de Génétique ...

  4. Somatic Embryogenesis and Plant Regeneration in Eggplant ...

    African Journals Online (AJOL)

    DR SIDHU

    2013-02-20

    Feb 20, 2013 ... Two as well as three way interactions of three eggplant genotypes, media compositions and explants. (hypocotyl, cotyledon and ... hypocotyl of BR-16 was not able to induce somatic embryogenesis on MS media fortified with 1.5 mgl-1 IBA + 1.0 mgl-1 BAP. ..... Manipulation of Plants. Oxford University Press ...

  5. PLANT REGENERATION THROUGH TISSUE CULTURE OF PEAR ...

    African Journals Online (AJOL)

    AISA

    PEAR MILLET (Pennisetum Glaucum (L) R.) K.TIECOURA 1, L. LEDOUX.2 AND M. DINANT.2. 1 Laboratoire de Génétique et amélioration des plantes, UFR de Biosciences, Université de Cocody,. B.P.582 Abidjan 22, Côte d'Ivoire. 2 Laboratoire de Génétique Moléculaire, Département de Botanique, Université de Liège, ...

  6. High frequency plant regeneration from mature seedderived callus ...

    African Journals Online (AJOL)

    In the present study, we have developed a high-frequency plant regeneration system for Italian ryegrass via callus culture using mature seeds as explants. Optimal embryogenic callus induction was found to occur in MS medium containing 5 mg l-1 2,4-D, 0.5 mg l-1 BA, 500 mg l-1 L-proline, 1 g l-1 casein hydrolysate, 30 g ...

  7. High frequency plant regeneration from mature seed- derived callus ...

    African Journals Online (AJOL)

    PRECIOUS

    2009-12-15

    Dec 15, 2009 ... In the present study, we have developed a high-frequency plant regeneration system for Italian ryegrass via callus culture using mature seeds as explants. Optimal embryogenic callus induction was found to occur in MS medium containing 5 mg l-1 2,4-D, 0.5 mg l-1 BA, 500 mg l-1 L-proline, 1 g l-1 casein.

  8. Catharanthus roseus (L. G. Don - plant regeneration and alkaloids content

    Directory of Open Access Journals (Sweden)

    Mirosława Furmanowa

    2014-01-01

    Full Text Available We describe here a regeneration of plantlets of Catharanthus roseus (L. G. Don from shoot tips and axillary buds. Shoot tips were excised from 7-day-old seedlings and were incubated in solid Nitsch and Nitsch (NN medium supplemented with kinetin, benzyladenine (BA, indole-3-butyric acid (IBA and β-indolylacetic acid (IAA in various combinations. After two months in culture, regenerated rooted plantlets were cut and transferred to a new medium; the explants contained shoot tips or axillary buds. Four passages were done. We obtained about 200 rooting plantlets from one seedling. Then the plantlets were transferred to the soil and they grew under a foil tent. After five months of vegetation they were collected, dried and weighed. Chemical investigations of leaves of these plants were done. The vindoline and catharanthine were dominant alkaloids in the juvenile stage of plants (before blooming. Total amount of alkaloids, equal 2.95%, was gravimetrically determined in leaves of plants, after 4th passage, regenerated in vitro on NN medium supplemented with kinetin and IBA.

  9. [Plant regeneration and clonal propagation system of Datura metel].

    Science.gov (United States)

    Wang, Feng-Ying; Sun, Yi-Ming; Zhang, Hong; Cheng, Meng-Qi; Zhang, Lai; Sun, Min

    2014-02-01

    To study the condition of plant regeneration and clonal propagation system of Datura metel. Stems and leaves of Datura metel were used as explants, effects of different hormones for callus induction and plant regeneration of leaves and clonal propagation system of stems were studied and optimized. The optimal way to obtain sterile explant for leaves were sterilized in 75% ethyl alcohol for 6 s then 0.1% HgCl2 for 6 min; Stems were sterilized in 75% ethyl alcohol for 8 s then 0.1% HgCl2 for 7 min. The optimal medium for callus of leaves was MS + 1.0 mg/L 6-BA + 0.1 mg/L NAA; The optimal medium for callus induction of clustered buds was MS + 2.0 mg/L 6-BA + 0.2 mg/L NAA; The optimal medium for clonal propagation system of stems was MS + 3.0 mg/L 6-BA + 0.05 mg/L NAA. The best medium for rooting induction was MS + 0.5 mg/L IBA. Transplant survival rate of plantlet was greater than 90% in humus soil-pearlite (5:1). The condition of plant regeneration and clonal propagation system of Datura metel is established.

  10. Plants for water recycling, oxygen regeneration and food production

    Science.gov (United States)

    Bubenheim, D. L.

    1991-01-01

    During long-duration space missions that require recycling and regeneration of life support materials the major human wastes to be converted to usable forms are CO2, hygiene water, urine and feces. A Controlled Ecological Life Support System (CELSS) relies on the air revitalization, water purification and food production capabilities of higher plants to rejuvenate human wastes and replenish the life support materials. The key processes in such a system are photosynthesis, whereby green plants utilize light energy to produce food and oxygen while removing CO2 from the atmosphere, and transpiration, the evaporation of water from the plant. CELSS research has emphasized the food production capacity and efforts to minimize the area/volume of higher plants required to satisfy all human life support needs. Plants are a dynamic system capable of being manipulated to favour the supply of individual products as desired. The size and energy required for a CELSS that provides virtually all human needs are determined by the food production capacity. Growing conditions maximizing food production do not maximize transpiration of water; conditions favoring transpiration and scaling to recycle only water significantly reduces the area, volume, and energy inputs per person. Likewise, system size can be adjusted to satisfy the air regeneration needs. Requirements of a waste management system supplying inputs to maintain maximum plant productivity are clear. The ability of plants to play an active role in waste processing and the consequence in terms of degraded plant performance are not well characterized. Plant-based life support systems represent the only potential for self sufficiency and food production in an extra-terrestrial habitat.

  11. Exotic weeds and fluctuating microclimate can constrain native plant regeneration in urban forest restoration.

    Science.gov (United States)

    Wallace, K J; Laughlin, Daniel C; Clarkson, Bruce D

    2017-06-01

    thresholds in some ecosystem properties makes conditions suitable for the regeneration of late successional species, which is vital for restoration success and long-term ecosystem sustainability. Abiotic and biotic conditions that promote tree regeneration and epiphyte colonization will likely be present in forests with a basal area ≥27 m2 /ha. We recommend that urban forest restoration plantings be designed to promote rapid canopy closure to reduce light availability, suppress herbaceous weeds, and stabilize the microclimate. © 2017 by the Ecological Society of America.

  12. Plant regeneration from protoplast of Brazilian citrus cultivars

    Directory of Open Access Journals (Sweden)

    GLORIA FERNANDA JANUZZI MENDES DA

    2000-01-01

    Full Text Available A procedure is described to regenerate plants from protoplasts of Brazilian citrus cultivars, after isolation, fusion and culture. Protoplasts were isolated from embryogenic cell suspension cultures and from leaf mesophyll of seedlings germinated in vitro. The enzyme solution for protoplast isolation was composed of mannitol (0.7 M, CaCl2 (24.5 mM, NaH2PO4 (0.92 mM, MES (6.15 mM, cellulase (Onozuka RS - Yakult, 1%, macerase (Onozuka R10 - Yakult, 1% and pectolyase Y-23 (Seishin, 0.2%. Protoplast culture in liquid medium after chemical fusion lead to the formation of callus colonies further adapted to solid medium. Somatic embryo formation occurred spontaneously after two subcultures, on modified MT medium supplemented with 500 mg/L of malt extract. Well defined embryos were germinated in modified MT medium with addition of GA3 (2.0 muM and malt extract (500 mg/L. Plant regeneration was also achieved by adventitious shoots obtained through direct organogenesis of not well defined embryos in modified MT medium with addition of malt extract (500 mg/L, BAP (1.32 muM, NAA (1.07 muM and coconut water (10 mL/L. Plantlets were transferred to root medium. Rooted plants were transferred to a greenhouse for further adaptation and development.

  13. Enrichment of By-Product Materials from Steel Pickling Acid Regeneration Plants (TRP 9942)

    Energy Technology Data Exchange (ETDEWEB)

    Lu Swan, Delta Ferrites LLC

    2009-09-30

    A new process for manufacturing an enriched, iron-based product (strontium hexaferrite) in existing steel pickling acid regeneration facilities was evaluated. Process enhancements and equipment additions were made to an existing acid regeneration plant to develop and demonstrate (via pilot scale testing and partial-capacity production trials) the viability of a patented method to produce strontium-based compounds that, when mixed with steel pickling acid and roasted, would result in a strontium hexaferrite powder precursor which could then be subjected to further heat treatment in an atmosphere that promotes rapid, relatively low-temperature formation of discrete strontium hexaferrite magnetic domains yielding an enriched iron-based product, strontium hexaferrite, that can be used in manufacturing hard ferrite magnets.

  14. Plant regeneration from in vitro leaves of mature black cherry (Prunus serotina)

    Science.gov (United States)

    Xiaomei Liu; Paula M. Pijut

    2008-01-01

    A regeneration system was developed for Prunus serotina from a juvenile (F) and two mature genotypes (#3 and #4). Adventitious shoots regenerated from leaves of in vitro cultures on woody plant medium with thidiazuron (TDZ) and naphthaleneacetic acid (NAA). The best regeneration for genotype F (91.4%) was observed on medium with 9.08 µM TDZ...

  15. Adventitious shoot formation and plant regeneration from leaf ...

    African Journals Online (AJOL)

    High root regeneration was obtained when the explants were cultured on medium with NAA only. Shoot regeneration was associated with callus formation. Regenerated shoots were rooted ex vitro, acclimatized and grown normally in the greenhouse. Keywords: Cytokinins, carnation, multiplication, regeneration, thidiazuron

  16. Gene-modified stem cells combined with rapid prototyping techniques: a novel strategy for periodontal regeneration.

    Science.gov (United States)

    He, Huixia; Cao, Junkai; Wang, Dongsheng; Gu, Bing; Guo, Hong; Liu, Hongchen

    2010-03-01

    Periodontal disease, a worldwide prevalent chronic disease in adults, is characterized by the destruction of the periodontal supporting tissue including the cementum, periodontal ligament and alveolar bone. The regeneration of damaged periodontal tissue is the main goal of periodontal treatment. Because conventional periodontal treatments remain insufficient to attain complete and reliable periodontal regeneration, periodontal tissue engineering has emerged as a prospective alternative method for improving the regenerative capacity of periodontal tissue. However, the potential of periodontal regeneration seems to be limited by the understanding of the cellular and molecular events in the formation of periodontal tissue and by the insufficient collaboration of multi-disciplinary research that periodontal tissue engineering involves. In this paper, we first reviewed the recent advancements in stem cells, signaling factors, and scaffolds that relate to periodontal regeneration. Then we speculate that specific genes would improve regenerative capacity of these stem cells, which could differentiate into cementoblasts, osteoblasts and fibroblasts. In addition, the 3D scaffolds that mimic the different structure and physiologic functions of natural fibro-osseous tissue could be fabricated by rapid prototyping (RP) techniques. It was therefore hypothesized that gene-modified stem cells combined with rapid prototyping techniques would be a new strategy to promote more effective and efficient periodontal regeneration.

  17. Enhanced plant regeneration in grain and sweet sorghum by asparagine, proline and cefotaxime.

    Science.gov (United States)

    Rao, A M; Sree, K P; Kishor, P B

    1995-01-01

    Cefotaxime ( 50 and 100 mg/1 ), a cephalosporin antibiotic and the amino acids asparagine and proline (200 mg/l) enhanced the production of embryogenic callus, increased the frequency of plant regeneration, and delayed the loss of regeneration potential in immature embryo-derived callus cultures ofSorghum bicolor (L.) Moench. Although these compounds did not promote callus induction or growth of callus, they influenced plant regeneration considerably in 10 low responding genotypes of grain and high anthocyanin containing sweet sorghums.

  18. Dynamics of planted cherrybark oak seedlings and yellow-poplar from seed following a regeneration clearcut

    Science.gov (United States)

    Wayne K. Clatterbuck

    2011-01-01

    A bottomland loblolly pine (Pinus taeda) stand originating from an early 1940s planting on a minor stream bottom of the Coastal Plain in west Tennessee was harvested in 1992 and allowed to regenerate to hardwoods. Although the pines had been planted, a few naturally regenerated yellow-poplar (Liriodendron tulipifera) and sweetgum...

  19. In vitro culture and plant regeneration derived from ray florets of ...

    African Journals Online (AJOL)

    STORAGESEVER

    2010-02-22

    Feb 22, 2010 ... significant differences test (LSD). The data were analyzed, using statistical analysis system (SAS) programme, version 6 (1985). RESULTS AND DISCUSSION. In vitro culture and plant regeneration of chrysanthemum cultivars. Establishing reliable in vitro plant regeneration is a pre- requisite step before ...

  20. Somaclonal variation in Asparagus officinalis plants regenerated by organogenesis from long-term callus cultures

    Directory of Open Access Journals (Sweden)

    Pontaroli Ana Clara

    2005-01-01

    Full Text Available Somaclonal variation in plants regenerated by organogenesis from long-term cultured calluses of two diploid staminate genotypes of Asparagus officinalis cv. Argenteuil was characterized by plant phenotype, ploidy, meiotic behavior, pollen viability, fruit and seed set, and AFLP profiles. Phenotypic deviations from the donors were detected in foliage color, flower size, and cladode and flower morphology. Ploidy changes were observed in 37.8% of the 37 regenerants studied. Meiotic alterations in 12 out of 21 regenerants included laggards, dicentric bridges, micronuclei, restitution nuclei and polyads. Of the 408 AFLP markers screened in 43 regenerants and the donors, 2.94% showed polymorphism. High pollen viability was observed in the 22 regenerants analyzed. All crosses between one pistillate plant and 35 regenerants, as well as the controls, produced fruits and seeds; however, no plump seeds resulted in 35.3% of the crosses with regenerants, and no seeds germinated in 12.5% of those with apparently normal seeds. Fruit and seed set was similar in crosses with diploid regenerants with normal meiosis and the controls but was lower in crosses with diploid and polyploid regenerants with abnormal meiosis. Our results show that the regenerated plants exhibited conspicuous somaclonal variation that could be eventually exploited for in vitro selection systems.

  1. Enhancing plant regeneration in tissue culture: a molecular approach through manipulation of cytokinin sensitivity.

    Science.gov (United States)

    Hill, Kristine; Schaller, G Eric

    2013-10-01

    Micropropagation is used for commercial purposes worldwide, but the capacity to undergo somatic organogenesis and plant regeneration varies greatly among species. The plant hormones auxin and cytokinin are critical for plant regeneration in tissue culture, with cytokinin playing an instrumental role in shoot organogenesis. Type-B response regulators govern the transcriptional output in response to cytokinin and are required for plant regeneration. In our paper published in Plant Physiology, we explored the functional redundancy among the 11 type-B Arabidopsis response regulators (ARRs). Interestingly, we discovered that the enhanced expression of one family member, ARR10, induced hypersensitivity to cytokinin in multiple assays, including callus greening and shoot induction of explants. Here we 1) discuss the hormone dependence for in vitro plant regeneration, 2) how manipulation of the cytokinin response has been used to enhance plant regeneration, and 3) the potential of the ARR10 transgene as a tool to increase the regeneration capacity of agriculturally important crop plants. The efficacy of ARR10 for enhancing plant regeneration likely arises from its ability to transcriptionally regulate key cytokinin responsive genes combined with an enhanced protein stability of ARR10 compared with other type-B ARRs. By increasing the capacity of key tissues and cell types to respond to cytokinin, ARR10, or other type-B response regulators with similar properties, could be used as a tool to combat the recalcitrance of some crop species to tissue culture techniques.

  2. Specific changes in rapidly transported proteins during regeneration of the goldfish optic nerve.

    Science.gov (United States)

    Benowitz, L I; Shashoua, V E; Yoon, M G

    1981-03-01

    Double labeling methods were used to identify changes in the complement of proteins synthesized in the retinal ganglion cells and transported down the optic nerve during the process of axonal regeneration. Eight to 62 days after goldfish underwent a unilateral optic nerve crush, one eye was labeled with [3H]-, the other with [14C]proline. Control and regenerating optic nerves were dissected out and homogenized together after 5 hr, a time which allowed us to examine selectively membrane-bound components which migrate in the rapid phase of axoplasmic transport. Proteins from the two sides were so-purified and separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Analysis of the 3H and 14C incorporation patterns along the gels revealed a radical shift away from the normal labeling spectrum during regeneration, with selective changes in labeling at particular molecular weights varying over a 3-fold range. Eight days after crushing the optic nerve, the greatest increases in labeling were seen for material with apparent molecular weights of 24,000 to 27,000, 44,000, and 210,000 daltons. These peaks declined thereafter, and on days 29 to 39, the most prominent increases were at 110,000 to 140,000 daltons. These studies indicate a continuously changing pattern in the synthesis and/or degradation of proteins that are rapidly transported down the optic nerve during regeneration and point to molecular species potential significance in the establishment of the visual map upon the brain.

  3. Specific changes in rapidly transported proteins during regeneration of the goldfish optic nerve

    Energy Technology Data Exchange (ETDEWEB)

    Benowitz, L.I.; Shashoua, V.E.; Yoon, M.G.

    1981-03-01

    Double labeling methods were used to identify changes in the complement of proteins synthesized in the retinal ganglion cells and transported down the optic nerve during the process of axonal regeneration. Eight to 62 days after goldfish underwent a unilateral optic nerve crush, one eye was labeled with (3H)-, the other with (14C)proline. Control and regenerating optic nerves were dissected out and homogenized together after 5 hr, a time which allowed us to examine selectively membrane-bound components which migrate in the rapid phase of axoplasmic transport. Proteins from the two sides were so-purified and separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Analysis of the 3H and 14C incorporation patterns along the gels revealed a radical shift away from the normal labeling spectrum during regeneration, with selective changes in labeling at particular molecular weights varying over a 3-fold range. Eight days after crushing the optic nerve, the greatest increases in labeling were seen for material with apparent molecular weights of 24,000 to 27,000, 44,000, and 210,000 daltons. These peaks declined thereafter, and on days 29 to 39, the most prominent increases were at 110,000 to 140,000 daltons. These studies indicate a continuously changing pattern in the synthesis and/or degradation of proteins that are rapidly transported down the optic nerve during regeneration and point to molecular species potential significance in the establishment of the visual map upon the brain.

  4. Plant MAPK cascades: Just rapid signaling modules?

    KAUST Repository

    Boudsocq, Marie

    2015-08-27

    © 2015 Taylor & Francis Group, LLC. Abscisic acid (ABA) is a major phytohormone mediating important stress-related processes. We recently unveiled an ABA-activated MAPK signaling module constituted of MAP3K17/18-MKK3-MPK1/2/7/14. Unlike classical rapid MAPK activation, we showed that the activation of the new MAPK module is delayed and relies on the MAP3K protein synthesis. In this addendum, we discuss the role of this original and unexpected activation mechanism of MAPK cascades which suggests that MAPKs can regulate both early and longterm plant stress responses.

  5. Plant regeneration and ploidy variation in culture derived plants of Asclepias curassavica L.

    Science.gov (United States)

    Pramanik, T K; Datta, S K

    1986-06-01

    Clonal propagation of medicinal milkweed, Asclepias curassavica L. (Asclepiadaceae) was achieved by culturing excised nodes on MS medium (Murashige and Skoog, 1962) supplemented with different hormone combinations. Both BAP and Kn were found equally effective for shoot initiation. IAA and NAA were found suitable for root induction. Combinations of Kn and NAA induced both roots and shoots after 30 days of culture. Chromosomal variation was observed in the roots of in vitro regenerated plants. Regenerants with higher chromosome number (33; 2n=22) obtained on MS medium in response to 9.2 μM Kn+10.7 μM NAA showed vigorous growth and higher propagation rates in culture than the plants possessing less than the diploid chromosome number (2n-2=20, 2n-4=18). Such variations are more likely due to genetic fitness of different aneuploids grown on a particular nutrient medium.

  6. Callus induction and plant regeneration in the metallophyte Silene vulgaris (Caryophyllaceae).

    NARCIS (Netherlands)

    Jack, E.M.; Atanasova, S; Verkley, J.A.C.

    2005-01-01

    Zinc tolerant and non-tolerant ecotypes of Silene vulgaris (Moench) Garcke were examined for their suitability to provide an efficient and reproducible callus formation and regeneration system. Successful and rapid regeneration of adventitious shoots from callus was achieved in leaf tissue but not

  7. Plant regeneration from mesophyll protoplasts of Matthiola incana (L.) R. Br.

    Science.gov (United States)

    Siemens, J; Sacristán, M D

    1995-04-01

    A protocol for obtaining regenerated fertile plants from mesophyll protoplasts of three lines of Matthiola incana is described. Protoplasts were isolated from leaves of 21-28 days old Matthiola plants grown in controlled environment. Sustained divisions were achieved when protoplasts were embedded in sodium alginate. Up to 2.0 % of the protoplasts developed into colonies which could be transferred to shoot regeneration media. More than 25 % of the obtained calluses regenerated shoots. About 4 % of these shoots could be rooted and after transfer to soil phenotypically normal plants have been obtained.

  8. Somatic embryogenesis and plant regeneration in Wedelia calendulacea Less. an endangered medicinal plant

    OpenAIRE

    Shamima Akhtar Sharmin; Md. Jahangir Alam; Md. Mominul Islam Sheikh; Kanak Kanti Sarker; Muhammad Khalekuzzaman; Md. Anwarul Haque; Mohammad Firoz Alam; Iftekhar Alam

    2014-01-01

    In this work, plant regeneration via somatic embryogenesis was achieved from leaf and internode derived callus of Wedelia calendulacea, an endangered medicinal plant. Primary callus was induced by culturing leaf disc and internode explant on Murashige and Skoog medium supplemented with 2.0 mg L-1 of 2,4-D under light condition. Transfer of embryogenic callus on a reduced concentration of 2,4-D facilitated somatic embryo development while calluses remained unorganized at the same 2,4-D level. ...

  9. Plants regenerated from tissue culture contain stable epigenome changes in rice.

    Science.gov (United States)

    Stroud, Hume; Ding, Bo; Simon, Stacey A; Feng, Suhua; Bellizzi, Maria; Pellegrini, Matteo; Wang, Guo-Liang; Meyers, Blake C; Jacobsen, Steven E

    2013-03-19

    Most transgenic crops are produced through tissue culture. The impact of utilizing such methods on the plant epigenome is poorly understood. Here we generated whole-genome, single-nucleotide resolution maps of DNA methylation in several regenerated rice lines. We found that all tested regenerated plants had significant losses of methylation compared to non-regenerated plants. Loss of methylation was largely stable across generations, and certain sites in the genome were particularly susceptible to loss of methylation. Loss of methylation at promoters was associated with deregulated expression of protein-coding genes. Analyses of callus and untransformed plants regenerated from callus indicated that loss of methylation is stochastically induced at the tissue culture step. These changes in methylation may explain a component of somaclonal variation, a phenomenon in which plants derived from tissue culture manifest phenotypic variability. DOI:http://dx.doi.org/10.7554/eLife.00354.001.

  10. Pollination and seed dispersal are the most threatened processes of plant regeneration

    Science.gov (United States)

    Neuschulz, Eike Lena; Mueller, Thomas; Schleuning, Matthias; Böhning-Gaese, Katrin

    2016-07-01

    Plant regeneration is essential for maintaining forest biodiversity and ecosystem functioning, which are globally threatened by human disturbance. Here we present the first integrative meta-analysis on how forest disturbance affects multiple ecological processes of plant regeneration including pollination, seed dispersal, seed predation, recruitment and herbivory. We analysed 408 pairwise comparisons of these processes between near-natural and disturbed forests. Human impacts overall reduced plant regeneration. Importantly, only processes early in the regeneration cycle that often depend on plant-animal interactions, i.e. pollination and seed dispersal, were negatively affected. Later processes, i.e. seed predation, recruitment and herbivory, showed overall no significant response to human disturbance. Conserving pollination and seed dispersal, including the animals that provide these services to plants, should become a priority in forest conservation efforts globally.

  11. Rapid release of growth factors regenerates force output in volumetric muscle loss injuries

    Science.gov (United States)

    Grasman, Jonathan M.; Do, Duc M.; Page, Raymond L.; Pins, George D.

    2015-01-01

    A significant challenge in the design and development of biomaterial scaffolds is to incorporate mechanical and biochemical cues to direct organized tissue growth. In this study, we investigated the effect of hepatocyte growth factor (HGF) loaded, crosslinked fibrin (EDCn-HGF) microthread scaffolds on skeletal muscle regeneration in a mouse model of volumetric muscle loss (VML). The rapid, sustained release of HGF significantly enhanced the force production of muscle tissue 60 days after injury, recovering more than 200% of the force output relative to measurements recorded immediately after injury. HGF delivery increased the number of differentiating myoblasts 14 days after injury, and supported an enhanced angiogenic response. The architectural morphology of microthread scaffolds supported the ingrowth of nascent myofibers into the wound site, in contrast to fibrin gel implants which did not support functional regeneration. Together, these data suggest that EDCn-HGF microthreads recapitulate several of the regenerative cues lost in VML injuries, promote remodeling of functional muscle tissue, and enhance the functional regeneration of skeletal muscle. Further, by strategically incorporating specific biochemical factors and precisely tuning the structural and mechanical properties of fibrin microthreads, we have developed a powerful platform technology that may enhance regeneration in other axially aligned tissues. PMID:26344363

  12. Efficient plant regeneration of bittersweet (Solanum dulcamara L., a medicinal plant

    Directory of Open Access Journals (Sweden)

    Arzu Ucar Turker

    2011-01-01

    Full Text Available Solanum dulcamara L. (bittersweet is a medicinal plant that has been used to treat skin diseases, warts, tumors, felons, arthritis, rheumatism, bronchial congestion, heart ailments, ulcerative colitis, eye inflammations, jaundice and pneumonia. A reliable in vitro culture protocol for bittersweet was established. Explants (leaf and petiole segments were cultured on Murashige and Skoog minimal organics (MSMO medium with various plant growth regulator combinations. Leaf explants formed more shoots than petiole explants. Plant regeneration was observed through indirect organogenesis with both explants. Best shoot proliferation was obtained from leaf explants with 3 mg/l BA (benzyladenine and 0.5 mg/l IAA (indole-3-acetic acid. Regenerated shoots were transferred to rooting media containing different levels of IAA (indole-3-acetic acid, IBA (indole-3-butyric acid, NAA (naphthalene acetic acid or 2,4-D (2,4 dichlorophenoxyacetic acid. Most shoots developed roots on medium with 0.5 mg/l IBA. Rooted explants were transferred to vermiculate in Magenta containers for acclimatization and after 2 weeks, they were planted in plastic pots containing potting soil and maintained in the plant growth room.

  13. Effect of sorbitol in callus induction and plant regeneration in wheat

    African Journals Online (AJOL)

    STORAGESEVER

    2009-12-01

    Dec 1, 2009 ... ples of such achievements is green revolution, which resulted in ... activities was focused on in vitro culture and regeneration as a tool of cereal breeding in the recent years. It is well documented that the genetic engineering of cereals currently depends on the use of tissue culture and plant regeneration ...

  14. An efficient plant regeneration and Agrobacterium-mediated genetic transformation of Tagetes erecta.

    Science.gov (United States)

    Gupta, Vijayta; Ur Rahman, Laiq

    2015-07-01

    Tagetes erecta, L. an asteraceous plant of industrial and medicinal value, contains important compounds like pyrethrins, thiophenes and lutein, possessing immense potential for insecticidal, nematicidal and nutraceutical activities. Considering the importance and demand for these natural compounds, genetic manipulation of this crop for better productivity of secondary metabolites holds great significance. A rapid and reproducible direct regeneration and genetic transformation system is the prerequisite for genetic manipulation of any crop. This paper elucidates the establishment of an efficient direct regeneration and transformation protocol of T. erecta using Agrobacterium tumefaciens. Investigation of the effects of different types of explants (Hypocotyls, cotyledonary leaves, rachis and leaf sections) and different BAP and GA3 combinations on the regeneration frequency of T. erecta suggested that the best regeneration frequency (66 %) with an average of 5.08 ± 0.09 shoot buds/explant was observed from hypocotyl explants cultured on media containing 1.5 mg/l BAP and 5 mg/l GA3. The transformation protocol was established using A. tumefaciens strain LBA4404, containing the binary vector pBI121, along with the gusA reporter gene with intron under the transcriptional control of the Cauliflower Mosaic Virus (CaMV) 35S promoter and the neomycin phosphotransferase II (nptII) gene as a kanamycin-resistant plant-selectable marker. Various parameters like optimization of kanamycin concentration (200 mg/l) for selection, standardization of cocultivation time (45 min) and acetosyringone concentration (150 μM) for obtaining higher transformation frequency were established using hypocotyl explants. The selected putative transgenic shoots were subsequently rooted on the Murashige and Skoog medium and transferred to the green house successfully. The plants were characterised by analysing the gus expression, amplification of 600 bp npt II fragment and Southern blot

  15. Somatic embryogenesis and plant regeneration in Anthurium andraeanum hybrids.

    Science.gov (United States)

    Kuehnle, A R; Chen, F C; Sugii, N

    1992-08-01

    A method for the production of somatic embryos and subsequent plant regeneration for Anthurium andraeanum Linden ex André (Monocotyledonae) hybrids is described. Whole leaf blade explants, derived from plantlets grown in vitro, formed translucent embryogénic calli at their basal ends within one month of culture in the dark. Secondary somatic embryos formed frequently and without an intervening callus on surfaces of primary embryos. Embryogenesis was induced with three genotypes using a modified half-strength Murashige and Skoog (MS) medium supplemented with 1.0 to 4.0 mg l(-1) 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.33 to 1.0 mg l(-1) kinetin. A combination of 2% sucrose with 1% glucose in the medium favored embryogenesis over 3% sucrose alone. Whole leaf blades on medium solidified with 0.18% Gelrite produced more somatic embryos than leaves on medium with 0.7% Bacto-agar. Within two to three months after culture initiation, embryos were transferred to modified MS medium containing 0.2 mg l(-1) 6-benzyladenine (BA) and 2% sucrose and placed in the light for conversion into plantlets. Rooted plantlets were recovered and transferred into pots with tree fern fiber medium and grown in the greenhouse.

  16. Regeneration of plants from Fraxinus pennsylvanica hypocotyls and cotyledons

    Science.gov (United States)

    Ningxia Du; Paula M. Pijut

    2008-01-01

    An adventitious shoot regeneration and rooting protocol was developed for green ash (Fraxinus pennsylvanica) seedling explants. The best regeneration medium for freshly isolated hypocotyls and cotyledons was Murashige and Skoog (MS) supplemented with 13.3 µM6-benzylaminopurine (BA) plus 4.5 µM thidiazuron (TDZ), and 22.2 µM...

  17. Factors influencing callus induction and plant regeneration of ...

    African Journals Online (AJOL)

    Establishment of highly efficient and reproducible regeneration system would greatly influence the efforts of improvement of Elymus sp. through gene transfer technology. A suitable callus induction and efficient regeneration protocol for Dahurian wildrye grass (Elymus dahuricus L.) was developed. It consisted of 3.0 mg/L 2 ...

  18. Optimization of a plant regeneration protocol for broccoli | Huang ...

    African Journals Online (AJOL)

    The factors which influence the regeneration of broccoli (Brassica oleracea var. italica) were studied using an orthogonal design. The results showed that the major factor was the explant type, followed by naphthylacetic acid (NAA), benzylaminopurine (BAP), sucrose and AgNO3 in turn. The maximum regeneration was on ...

  19. Production of new 3D scaffolds for bone tissue regeneration by rapid prototyping.

    Science.gov (United States)

    Fradique, R; Correia, T R; Miguel, S P; de Sá, K D; Figueira, D R; Mendonça, A G; Correia, I J

    2016-04-01

    The incidence of bone disorders, whether due to trauma or pathology, has been trending upward with the aging of the worldwide population. The currently available treatments for bone injuries are rather limited, involving mainly bone grafts and implants. A particularly promising approach for bone regeneration uses rapid prototyping (RP) technologies to produce 3D scaffolds with highly controlled structure and orientation, based on computer-aided design models or medical data. Herein, tricalcium phosphate (TCP)/alginate scaffolds were produced using RP and subsequently their physicochemical, mechanical and biological properties were characterized. The results showed that 60/40 of TCP and alginate formulation was able to match the compression and present a similar Young modulus to that of trabecular bone while presenting an adequate biocompatibility. Moreover, the biomineralization ability, roughness and macro and microporosity of scaffolds allowed cell anchoring and proliferation at their surface, as well as cell migration to its interior, processes that are fundamental for osteointegration and bone regeneration.

  20. Development of efficient plant regeneration and transformation system for impatiens using Agrobacterium tumefaciens and multiple bud cultures as explants.

    Science.gov (United States)

    Dan, Yinghui; Baxter, Aaron; Zhang, Song; Pantazis, Christopher J; Veilleux, Richard E

    2010-08-09

    has the advantages of 1) efficient, simple and rapid regeneration and transformation (with no need for sterilization or a greenhouse to grow stock plants), 2) flexibility (available all the time) for in vitro manipulation, 3) uniform and desirable green tissue explants for both nuclear and plastid transformation using Agrobacterium-mediated and biolistics methods, 4) no somaclonal variation and 5) resolution of necrosis of Agrobacterium-inoculated tissues.

  1. Plant regeneration from petiole and leaf of African violet (Saintpaulia ionantha Wendl. cultured in vitro

    Directory of Open Access Journals (Sweden)

    Wichada Sunpui

    2002-07-01

    Full Text Available A regeneration procedure was developed for in vitro grown African violet (Saintpaulia ionantha Wendl.. Adventitious shoot regeneration was compared among petioles, leaves and stems cultured on MS basal medium. High frequency regeneration was obtained from leaves and petioles supplemented with plant growth regulators (BA, KN, 2,4-D, and NAA. Regeneration media that induced the highest number of shoots in petiole explants was either MS medium supplemented with 3 mg/l BA alone or 1 mg/l NAA plus 3 mg/l BA. Adventitious shoots were also regenerated from callus that was derived from leaf explants cultured on MS medium containing several concentrations of TDZ. Callus induced on medium with 0.5 mg/l TDZ showed the best shoot induction. Regenerated shoots could be rooted on 1/2 MS medium or MS medium supplemented with 3 mg/l IBA. Complete plantlets were acclimatized and successfully transplanted to glasshouse conditions.

  2. Regeneration and growth rates of allofragments in four common stream plants

    DEFF Research Database (Denmark)

    Riis, Tenna; Madsen, Tom Vindbæk; Sennels, R. S. H.

    2009-01-01

    Colonisation by stream plants occurs to a large extent from simple stem fragments. Allofragments are stem fragments formed by mechanical breakage. We studied regeneration, colonisation, and growth rates in four common stream plants: Elodea canadensis Michx., Myriophyllum spicatum L., Potamogeton...... perfoliatus L. and Ranunculus baudotii x pseudofluitans. The objectives of this study were to determine (1) if shoots with an apical tip have higher regeneration (growth of new shoots and rhizomes from allofragments) and colonisation (root attachment in sediment) abilities and higher relative growth rates...... (RGR) than shoots without an apical tip, and (2) if fragment size correlates with regeneration and colonisation abilities and with RGR of fragments. For all species, over 60% of fragments regenerated new shoots and colonised. Apical shoots and larger fragments generally had higher regeneration...

  3. Callus Induction and Plant Regeneration from Mature Embryos of Oat (Avena sativa L.)

    OpenAIRE

    BİRSİN, Melahat A.

    2001-01-01

    The aim of this study, which was conducted in Ankara University, Faculty of Agriculture, Department of Field Crops, biotechnology laboratory in 1998-1999, was to determine the callus induction and plant regeneration capacity of oat (Avena sativa L.). The mature embryos of 10 oat genotypes were utilised for callus induction. The statistical analysis of the results revealed that callus induction and plant regeneration capacity were dependent on the oat genotype. However, a significant relations...

  4. An Efficient Plant Regeneration and Transformation System of Ma Bamboo (Dendrocalamus latiflorus Munro) Started from Young Shoot as Explant

    Science.gov (United States)

    Ye, Shanwen; Cai, Changyang; Ren, Huibo; Wang, Wenjia; Xiang, Mengqi; Tang, Xiaoshan; Zhu, Caiping; Yin, Tengfei; Zhang, Li; Zhu, Qiang

    2017-01-01

    Genetic engineering technology has been successfully used in many plant species, but is limited in woody plants, especially in bamboos. Ma bamboo (Dendrocalamus latiflorus Munro) is one of the most important bamboo species in Asia, and its genetic improvement was largely restricted by the lack of an efficient regeneration and transformation method. Here we reported a plantlet regeneration and Agrobacterium-mediated transformation protocol by using Ma bamboo young shoots as explants. Under our optimized conditions, embryogenic calluses were successfully induced from the excised young shoots on callus induction medium and rapidly grew on callus multiplication medium. Shoots and roots were regenerated on shoot induction medium and root induction medium, respectively, with high efficiency. An Agrobacterium-mediated genetic transformation protocol of Ma bamboo was established, verified by PCR and GUS staining. Furthermore, the maize Lc gene under the control of the ubiquitin promoter was successfully introduced into Ma bamboo genome and generated an anthocyanin over-accumulation phenotype. Our methods established here will facilitate the basic research as well as genetic breeding of this important bamboo species. Key achievements: A stable and high efficiency regeneration and Agrobacterium-mediated transformation protocol for Ma bamboo from vegetative organ is established. PMID:28798758

  5. Localization of QTLs for in vitro plant regeneration in tomato

    Directory of Open Access Journals (Sweden)

    Nuez Fernando

    2011-10-01

    Full Text Available Abstract Background Low regeneration ability limits biotechnological breeding approaches. The influence of genotype in the regeneration response is high in both tomato and other important crops. Despite the various studies that have been carried out on regeneration genetics, little is known about the key genes involved in this process. The aim of this study was to localize the genetic factors affecting regeneration in tomato. Results We developed two mapping populations (F2 and BC1 derived from a previously selected tomato cultivar (cv. Anl27 with low regeneration ability and a high regeneration accession of the wild species Solanum pennellii (PE-47. The phenotypic assay indicated dominance for bud induction and additive effects for both the percentage of explants with shoots and the number of regenerated shoots per explant. Two linkage maps were developed and six QTLs were identified on five chromosomes (1, 3, 4, 7 and 8 in the BC1 population by means of the Interval Mapping and restricted Multiple QTL Mapping methods. These QTLs came from S. pennellii, with the exception of the minor QTL located on chromosome 8, which was provided by cv. Anl27. The main QTLs correspond to those detected on chromosomes 1 and 7. In the F2 population, a QTL on chromosome 7 was identified on a similar region as that detected in the BC1 population. Marker segregation distortion was observed in this population in those areas where the QTLs of BC1 were detected. Furthermore, we located two tomato candidate genes using a marker linked to the high regeneration gene: Rg-2 (a putative allele of Rg-1 and LESK1, which encodes a serine/threonine kinase and was proposed as a marker for regeneration competence. As a result, we located a putative allele of Rg-2 in the QTL detected on chromosome 3 that we named Rg-3. LESK1, which is also situated on chromosome 3, is outside Rg-3. In a preliminary exploration of the detected QTL peaks, we found several genes that may be related

  6. Localization of QTLs for in vitro plant regeneration in tomato.

    Science.gov (United States)

    Trujillo-Moya, Carlos; Gisbert, Carmina; Vilanova, Santiago; Nuez, Fernando

    2011-10-20

    Low regeneration ability limits biotechnological breeding approaches. The influence of genotype in the regeneration response is high in both tomato and other important crops. Despite the various studies that have been carried out on regeneration genetics, little is known about the key genes involved in this process. The aim of this study was to localize the genetic factors affecting regeneration in tomato. We developed two mapping populations (F2 and BC1) derived from a previously selected tomato cultivar (cv. Anl27) with low regeneration ability and a high regeneration accession of the wild species Solanum pennellii (PE-47). The phenotypic assay indicated dominance for bud induction and additive effects for both the percentage of explants with shoots and the number of regenerated shoots per explant. Two linkage maps were developed and six QTLs were identified on five chromosomes (1, 3, 4, 7 and 8) in the BC1 population by means of the Interval Mapping and restricted Multiple QTL Mapping methods. These QTLs came from S. pennellii, with the exception of the minor QTL located on chromosome 8, which was provided by cv. Anl27. The main QTLs correspond to those detected on chromosomes 1 and 7. In the F2 population, a QTL on chromosome 7 was identified on a similar region as that detected in the BC1 population. Marker segregation distortion was observed in this population in those areas where the QTLs of BC1 were detected. Furthermore, we located two tomato candidate genes using a marker linked to the high regeneration gene: Rg-2 (a putative allele of Rg-1) and LESK1, which encodes a serine/threonine kinase and was proposed as a marker for regeneration competence. As a result, we located a putative allele of Rg-2 in the QTL detected on chromosome 3 that we named Rg-3. LESK1, which is also situated on chromosome 3, is outside Rg-3. In a preliminary exploration of the detected QTL peaks, we found several genes that may be related to regeneration. In this study we have

  7. Plant regeneration from leaf protoplasts of Solanum torvum.

    Science.gov (United States)

    Guri, A; Volokita, M; Sink, K C

    1987-07-01

    A protocol to obtain regenerated plants from protoplasts of Solanum torvum Sw a wild species of eggplant resistant to Verticillium wilt is reported. Leaf protoplasts were enzymatically isolated from six-week old seedlings grown in a controlled environment chamber. Protoplasts were plated on modified KM medium (0.4 M glucose)+(mg/l): 1.0 p-chlorophenoxyacetic acid (CPA)+1.0 naphthaleneacetic acid (NAA)+0.5 6-benzylaminopurine (BAP) and 0.02 abscisic acid (ABA). The protoplast density was 5×10(4) per ml with 5 ml placed in each of two quadrants in X-dishes (100×15 mm). The reservoir medium was modified KM+(mg/l): 0.1 NAA+0.5 BAP+0.1 M sucrose+0.1 M mannitol+0.6% washed agar+1% activated charcoal. Dishes were initially placed in the dark at 27°C. Protoplast division was initiated in 1-2 weeks and 4 weeks later p-calli were 1-3 mm. Plating efficiency was 11% when measured at 3 weeks. Six-week old p-calli were transferred individually onto Whatman No. 1 filter paper layered on modified KM (0.15 M sucrose)+mg/l: 2.0 indoleacetic acid (IAA)+2.0 zeatin+0.5% washed agar for 2 weeks. Subsequently, shoots occurred within 4 weeks at 70% efficiency on MS+30 g/l sucrose+2 mg/l zeatin. Shoots were rooted on half strength MS+10 g/l sucrose.

  8. Plant regeneration from petiole segments of some species in tissue culture

    Directory of Open Access Journals (Sweden)

    Krystyna Klimaszewska

    2013-12-01

    Full Text Available The regeneration ability of 21 plant species belonging to 14 families was tested. The method of tissue culture in vitro was applied, on basic MS medium with an addition of growth regulators from the auxin and cytokinin groups. From among the investigated plant groups Peperomia scandens and Caladium × hortulanum were capable of plant regeneration, Passiilora coerulea regenerated shoots, Hedera helix, Begonia glabra, Coleus blumei, Fuchsia hybrida, Passiflora suberosa and Peperomia eburnea formed callus and roots, Kalanchoe blossfeldiana, Pelargonium grandiflorum, P. peltatum, P. radula, Coleus shirensis and Magnolia soulangeana produced callus, Philodendron scandens, Rhododendron smirnovii, Hibiscus rosa-sinensis, Coprosma baueri, Cestrum purpureum and Solanum rantonnetii did not exhibit any regeneration reactions.

  9. Plant regeneration from immature embryos of Kenyan maize inbred ...

    African Journals Online (AJOL)

    SERVER

    2008-04-17

    0 - 20 mg L-1) and regenerated on hormone free MS ... for the purposes of genetically manipulating them to express phenotypes of ... excised and plated solid callus induction media with scutellar side in contact with media and ...

  10. Plant regeneration from mature embryo of commercial Indian bread wheat (Triticum aestivum L.) cultivars.

    Science.gov (United States)

    Parmar, Sanjay Singh; Sainger, Manish; Chaudhary, Darshna; Jaiwal, Pawan K

    2012-04-01

    A simple, efficient, reproducible and comparatively genotype-independent in vitro plant regeneration protocol was developed for ten commercial Indian bread wheat cultivars using mature embryos as the explants. Three different auxins and five different combinations of growth regulators in a modified Murashige and Skoog's basal medium were assessed for their effect on callus induction and plant regeneration, respectively, in a high yielding and widely grown cultivar, PBW-343. The optimized conditions were further evaluated with nine other commercial cultivars. A simple novel approach of physical isolation of regenerable calli from non regenerable structures during the early callus phase was used to improve plant regeneration. Callus induced on 2.0 mg(-1) 2,4-dichlorophenoxyacetic acid (2,4-D) showed a regeneration frequency of 86 % with 7.5 shoots per explants on hormone-free medium. A considerable improvement in the regeneration frequency (up to 97 %) and the average of shoots (19 shoots per explants) was obtained with a combination of thidiazuron (TDZ) and 2,4-D.

  11. A valued Indian medicinal plant – Begonia malabarica Lam. : Successful plant regeneration through various explants and field performance

    Directory of Open Access Journals (Sweden)

    Sevanan Rajeshkumar

    2009-05-01

    Full Text Available A cost-effective and efficient protocol has been described in the present work for large-scale and rapid in vitro propagation of a valuable medicinal herb Begonia malabarica Lam. (Begoniaceae by shoot auxillary-bud proliferation and organogenesis on MS medium supplemented with 6-benzylaminopurine (BA; 0.0-8.8 mg/l and indole-3-acetic acid (IAA; 0.0-2.88 mg/l at different concentrations, either alone or in combinations. Initiation of callus formation from the base of the leaf lamina was observed on MS supplemented with BA, IAA and adenine sulphate. Root induction on shoots was achieved on full strength MS with IAA/ indole-3-butyric acid (IBA at different concentrations. MS medium with 4.4 mg/l BA and 1.4 mg/l IAA elicited the maximum number of shoots (10 multiple shoots from nodal explants. Leaf-based callus differentiated into more than 28 shoots on MS with 150 mg/l adenine sulphate. The regenerated shoots were rooted on MS with 1.2 mg/l IBA within ten days. Almost 95% of the rooted shoots survived hardening when transferred to the field. The regenerated plants did not show any morphological change and variation in levels of secondary metabolites when compared with the mother stock. Thus, a reproduction of B. malabarica was established through nodal and leaf explants. This protocol can be exploited for conservation and commercial propagation of this medical plant in the Indian subcontinent and might be useful for genetic improvement programs.

  12. Agrobacterium tumefaciens mediated transformation of Orychophragmus violaceus cotyledon and regeneration of transgenic plants.

    Science.gov (United States)

    Zhou, J; Wei, Z; Xu, Z; Liu, S; Luo, P

    1996-01-01

    Excised cotyledons of Orychophragmus violaceus were used as explants for tissue culture. They were cultured on the MS medium supplemented with BA (3 mg/L) and NAA (0.2 mg/L). When the regenerated buds were 2 cm long, they were excised and transferred onto 1/2 MS medium with IBA (0.03 mg/L), then the whole plants were regenerated. The frequency of plant regeneration was 100%. Subsequently, the genetic transformation of O. violaceus was studied. After 2-3 days of cocultivation with Agrobacterium tumefaciens strain A208se (pTiT37, pROA93), the cotyledons were transferred onto the selection medium containing 25 mg/L Km and 250 mg/L Ap. After shoots emerged, they were excised and transferred onto the rooting medium containing 25 mg/L Km and 100 mg/L Cef. The roots were formed within 4-5 weeks. The whole plants were transplanted into pots and grew well. The frequency of plant regeneration was about 51%. The regenerated plants showed high enzymatic activities of beta-glucuronidase and neomycine phosphotransferase II. Southern blot analysis confirmed that NPTII gene had been stably integrated into the chromosomal genome of O. violaceus. The transformation frequency was 5.6%. The first transgenic plant of O. violaceus is being reported.

  13. High Frequency Plant Regeneration System from Transverse Thin Cell Layer Section of In vitro Derived ‘Nadia’ Ginger Microrhizome

    Directory of Open Access Journals (Sweden)

    Dikash Singh THINGBAIJAM

    2014-03-01

    Full Text Available An efficient and reproducible procedure is outlined for rapid in vitro multiplication of Zingiber officinale var. ‘Nadia’ through high frequency shoot proliferation from transverse thin cell layer (tTCL sections of in vitro derived microrhizome. In vitro derived microrhizome of size 500 μm in thickness was used as initial explants for induction of somatic embryos. Among the different phytohormones tested, tTCL explants shows maximum calli proliferation in medium containing 2 mg/L 2,4-Dichlorophenoxyacetic acid (88.30±0.11%. Reduced concentration of 2,4 Dichlorophenoxyacetic acid was supplemented with different cytokinins for regeneration of callus. Among the different medium tested, optimum redifferentiation of somatic embryos were observed in medium containing 0.2 mg/L 2,4 Dichlorophenoxyacetic acid and 6.0 mg/L BAP (141.08±0.25. Clump of regenerated plantlets were further subculture and transfer into microrhizome inducing medium containing high sucrose concentration (8%. Plantlets with well developed microrhizome were successfully acclimatized and eventually transferred to the field. The application of studying embryo section for regeneration of plants might be useful alternative to ginger improvement programme. Histological analysis showed formation of somatic embryos and regenerated adventitious shoot.

  14. Plant regeneration from leaf protoplasts of Brassica oleracea var. italica CV Green Comet broccoli.

    Science.gov (United States)

    Robertson, D; Earle, E D

    1986-02-01

    A procedure is described for regeneration of plants from leaf protoplasts of the hybrid broccoli cultivar, Green Comet (Brassica oleracea var italica). The totipotency of protoplasts isolated from plants regenerated from hypocotyl explants (GCR) was greater than that of protoplasts from plants grown directly from seed (GC). Using medium B developed by Pelletier et al (1983), division efficiencies greater than 70% were obtained in leaf protoplasts isolated from GCR. Approximately 1% of these protoplasts formed calli on solidified medium; 77% of the calli regenerated shoots. In contrast, protoplasts from seed-grown material showed a lower division efficiency (15-22%) and fewer protoplast-derived calli produced shoots. Some of the 178 protoplast-derived plants grown to maturity had variant phenotypes.

  15. Studies on plant regeneration and somaclonal variation in Saintpaulia ionantha Wendl. (African violet).

    Science.gov (United States)

    Daud, Norhayati; Taha, Rosna Mat; Hasbullah, Nor Azlina

    2008-05-01

    Efficient plant regeneration of Saintpaulia ionantha (African violet) has been obtained in the present study. MS medium supplemented with 1.0 mg L(-1) IAA and 2.0 mg L(-1) Zeatin resulted in 100% shoot regeneration and induced the highest number of shoots (average 15.0 +/- 0.8 shoots per explant) after being cultured for 8 weeks. The above hormone combination was optimum for shoot regeneration. Most of Saintpaulia ionantha plantlets derived from tissue culture system could be hardened and transferred to the greenhouse conditions with 84.0 +/- 1.6% success rate. However, regenerated plantlets of Saintpaulia ionantha (even after 12-months-old) failed to flower. Morphological characters of regenerated plantlets of Saintpaulia ionantha were observed and compared with in vivo (intact) plants. Regenerated plantlets showed some differences in morphological characters, such as height and leaf size, texture and colour, but the plantlets showed no variation in leaf arrangement and leaf margin. However, the morphological characters of the regenerated plantlets were found to be unstable.

  16. Crown expansion following thinning in naturally regenerated and planted longleaf pine

    Science.gov (United States)

    Steven B. Jack; Noah A. Jansen; Robert J. Mitchell

    2015-01-01

    The recent focus on restoration of longleaf pine (Pinus palustris Mill.) forests has frequently led to planting longleaf pine on old-field and cutover sites. While many perceptions regarding response of longleaf pine to management are based upon measurements in naturally regenerated stands, it is generally observed that crown development in planted longleaf stands is...

  17. Contrasting natural regeneration and tree planting in fourteen North American cities

    Science.gov (United States)

    David J. Nowak

    2012-01-01

    Field data from randomly located plots in 12 cities in the United States and Canada were used to estimate the proportion of the existing tree population that was planted or occurred via natural regeneration. In addition, two cities (Baltimore and Syracuse) were recently re-sampled to estimate the proportion of newly established trees that were planted. Results for the...

  18. In vitro plant regeneration of Esmeralda clarkei Rchb.f. via protocorm ...

    African Journals Online (AJOL)

    user

    2012-07-05

    Jul 5, 2012 ... with fragrant flowers. Reliable protocols for in vitro plant regeneration of E. clarkei via ... Plants blooms from fall to winter with three to four fragrant flowers. (Ghimire, 2008). It has high commercial value in ..... Cryopreservation of Dendrobium candidum. Wall. ex Lindl. Protocorm-like bodies by encapsulation.

  19. Effect of a low-level laser on bone regeneration after rapid maxillary expansion.

    Science.gov (United States)

    Cepera, Fernanda; Torres, Fernando C; Scanavini, Marco A; Paranhos, Luiz R; Capelozza Filho, Leopoldino; Cardoso, Mauricio A; Siqueira, Danieli C R; Siqueira, Danilo F

    2012-04-01

    In this study, we evaluated the effects of a low-level laser on bone regeneration in rapid maxillary expansion procedures. Twenty-seven children, aged 8 to 12 years, took part in the experiment, with a mean age of 10.2 years, divided into 2 groups: the laser group (n = 14), in which rapid maxillary expansion was performed in conjunction with laser use, and the no-laser group (n = 13), with rapid maxillary expansion only. The activation protocol of the expansion screw was 1 full turn on the first day and a half turn daily until achieving overcorrection. The laser type used was a laser diode (TWIN Laser; MMOptics, São Carlos, Brazil), according to the following protocol: 780 nm wavelength, 40 mW power, and 10 J/cm(2) density at 10 points located around the midpalatal suture. The application stages were 1 (days 1-5 of activation), 2 (at screw locking, on 3 consecutive days), 3, 4, and 5 (7, 14, and 21 days after stage 2). Occlusal radiographs of the maxilla were taken with the aid of an aluminum scale ruler as a densitometry reference at different times: T1 (initial), T2 (day of locking), T3 (3-5 days after T2), T4 (30 days after T3), and T5 (60 days after T4). The radiographs were digitized and submitted to imaging software (Image Tool; UTHSCSA, San Antonio, Tex) to measure the optic density of the previously selected areas. To perform the statistical test, analysis of covariance was used, with the time for the evaluated stage as the covariable. In all tests, a significance level of 5% (P maxillary expansion, provided efficient opening of the midpalatal suture and influenced the bone regeneration process of the suture, accelerating healing. Copyright © 2012 American Association of Orthodontists. Published by Mosby, Inc. All rights reserved.

  20. TDZ-induced plant regeneration in Astragalus cicer L.

    African Journals Online (AJOL)

    SERVER

    2008-04-17

    Apr 17, 2008 ... Adventitious root formation is very important for the vegetative propagation and a key step in micropropagated systems. Low frequency of rooting has been reported for some forage legumes such as sainfoin. (Pupilli et al., 1989) and alfalfa (Özgen et al., 1998) which appears to be a problem in regenerated ...

  1. Efficient plant regeneration from cotyledonary node explants of ...

    African Journals Online (AJOL)

    An efficient regeneration system was developed using cotyledonary node of two inbred line of melon, Cucumis melo 'CM-15' and 'CM-23'. The induction of shoots was achieved on Murashige and Skoog (MS) solid medium supplemented with different combinations of 6-benzylaminopurine (BA) and indole- 3-acetic acid ...

  2. TDZ-induced plant regeneration in Astragalus cicer L. | Basalma ...

    African Journals Online (AJOL)

    We developed a regeneration protocol using thidiazuron (TDZ) with a high frequency in vitro root induction in Astragalus cicer. High in vitro germination ratio (75%) for hard-seeds of A. cicer was also achieved. For this, hypocotyl and cotyledon explants were cultured on Murashige and Skoog medium supplemented with ...

  3. Optimization of callus induction and plant regeneration from ...

    African Journals Online (AJOL)

    An efficient regeneration system was developed using germinating seeds of two cultivars of sweet sorghum, Sorghum bicolor 'Yuantian No.1' and 'M81E', as explants. We tested different media supplements effects on callus induction. The effects of combinations of 2,4-D, KT, sucrose, agar and proline at different ...

  4. Regeneration of plants from Fraxinus nigra Marsh. hypocotyls

    Science.gov (United States)

    Rochelle R. Beasley; Paula M. Pijut

    2013-01-01

    Fraxinus nigra Marsh. (black ash) is a native North American hardwood tree species that is ecologically important and has ethnobotanical significance to American Indian communities of the eastern United States. Black ash has immature embryos at seed set, combined with complex stratification requirements, making natural regeneration difficult. This,...

  5. Improved embryogenic callus induction and plant regeneration in ...

    African Journals Online (AJOL)

    The objective of this study was to develop an efficient regeneration protocol in big bluestem, a potential feedstock that produces huge biomass. Embryogenic calli were induced from the seeds of cultivars, Kaw and Earl, on Murashige and Skoog (MS) medium with different concentrations of 2,4- dichlorophenoxyacetic acid (2 ...

  6. Establishment of a high-efficiency plant regeneration and ...

    African Journals Online (AJOL)

    further used to establish genetic transformation system, its embryogenic calli, initiated from immature zygotic embryos, were transformed with the plasmid p35SBarNos harboring Bar gene by microprojectile bombardment, after selection and differentiation culture, partial bombarded calli were regenerated into green plantlets ...

  7. Evaluation of somatic embryogenesis and plant regeneration in ...

    African Journals Online (AJOL)

    Optimization of tissue culture conditions for Sorghum bicolor L. through somatic embryogenesis from immature embryos is important for the genetic manipulation and improvement of this agronomically valuable crop. In an attempt to develop a successfully reproducible in vitro regeneration protocol for a group of diverse ...

  8. Plant regeneration in eggplant ( Solanum melongena L.): A review ...

    African Journals Online (AJOL)

    Eggplant is highly responsive to various tissue culture techniques. Somatic embryogenesis and direct organogenesis are widely studied protocols in this crop, but potential of regeneration varies with genotype, explant and culture media supplemented with different combination and concentration of growth hormones.

  9. Hairy root induction and plant regeneration of medicinal plant Dracocephalum kotschyi.

    Science.gov (United States)

    Sharafi, Ali; Sohi, Haleh Hashemi; Azadi, Pejman; Sharafi, Ata Allah

    2014-04-01

    An efficient hairy root induction system for an important endangered medicinal plant, Dracocephalum kotschyi, was developed through Agrobacterium rhizogenes-mediated transformation by modifying the co-cultivation medium using five bacterial strains, A4, ATCC15834, LBA9402, MSU440, and A13 (MAFF-02-10266). A drastic increase in transformation frequency was observed when a Murashige and Skoog medium lacking NH4NO3 KH2PO4, KNO3 and CaCl2 was used, resulting in hairy root induction frequencies of 52.3 %, 69.6 %, 48.6 %, 89.0 %, and 80.0 % by A4, A13, LBA9402, MSU440, and ATCC15834 strains, respectively. For shoot induction, hairy roots and unorganized tumors induced by strain ATCC15834 were placed on an MS media supplemented with 0.1, 0.25, 0.5, and 1 mg/l BA plus 0.1 mg/l NAA. The high frequency of shoot regeneration and number of shoot were obtained in the medium containing 0.25 mg/l BA and 0.1 mg/l NAA. Root induction occurred from the base of regenerated shoots on the MS medium supplemented with 0.5 mg/l IBA after 10 days.

  10. Derepression of the plant Chromovirus LORE1 induces germline transposition in regenerated plants.

    Directory of Open Access Journals (Sweden)

    Eigo Fukai

    2010-03-01

    Full Text Available Transposable elements represent a large proportion of the eukaryotic genomes. Long Terminal Repeat (LTR retrotransposons are very abundant and constitute the predominant family of transposable elements in plants. Recent studies have identified chromoviruses to be a widely distributed lineage of Gypsy elements. These elements contain chromodomains in their integrases, which suggests a preference for insertion into heterochromatin. In turn, this preference might have contributed to the patterning of heterochromatin observed in host genomes. Despite their potential importance for our understanding of plant genome dynamics and evolution, the regulatory mechanisms governing the behavior of chromoviruses and their activities remain largely uncharacterized. Here, we report a detailed analysis of the spatio-temporal activity of a plant chromovirus in the endogenous host. We examined LORE1a, a member of the endogenous chromovirus LORE1 family from the model legume Lotus japonicus. We found that this chromovirus is stochastically de-repressed in plant populations regenerated from de-differentiated cells and that LORE1a transposes in the male germline. Bisulfite sequencing of the 5' LTR and its surrounding region suggests that tissue culture induces a loss of epigenetic silencing of LORE1a. Since LTR promoter activity is pollen specific, as shown by the analysis of transgenic plants containing an LTR::GUS fusion, we conclude that male germline-specific LORE1a transposition in pollen grains is controlled transcriptionally by its own cis-elements. New insertion sites of LORE1a copies were frequently found in genic regions and show no strong insertional preferences. These distinctive novel features of LORE1 indicate that this chromovirus has considerable potential for generating genetic and epigenetic diversity in the host plant population. Our results also define conditions for the use of LORE1a as a genetic tool.

  11. Haploid technology allows for the efficient and rapid generation of homozygous antibody-accumulating transgenic tobacco plants.

    Science.gov (United States)

    Floss, Doreen M; Kumlehn, Jochen; Conrad, Udo; Saalbach, Isolde

    2009-09-01

    The large-scale production of plant-derived recombinant proteins requires the breeding of lines homozygous for the transgene(s). These can be selected by progeny testing over multiple sexual generations, but a more efficient means is to fix homozygosity in a single generation using doubled haploid technology. In this study, transgenic tobacco plants, hemizygous for both of the independently inherited genes encoding the light and heavy chains of the anti-human immunodeficiency virus monoclonal antibody 2F5, were used to establish embryogenic pollen cultures. The improved protocol employed in this study guaranteed a very high regeneration efficiency, with more than 50% of the regenerants being spontaneously doubled haploids. Hence, there was no requirement to chemically induce chromosome doubling to recover sufficient entirely homozygous recombinants. As expected, approximately 25% of the regenerants were homozygous for both transgenes. Thus, the employment of haploid technology allowed for the efficient and rapid generation of true-breeding tobacco lines accumulating functional immunoglobulins.

  12. Rapid neural circuit switching mediated by synaptic plasticity during neural morphallactic regeneration.

    Science.gov (United States)

    Lybrand, Zane R; Zoran, Mark J

    2012-09-01

    The aquatic oligochaete, Lumbriculus variegatus (Lumbriculidae), undergoes a rapid regenerative transformation of its neural circuits following body fragmentation. This type of nervous system plasticity, called neural morphallaxis, involves the remodeling of the giant fiber pathways that mediate rapid head and tail withdrawal behaviors. Extra- and intracellular electrophysiological recordings demonstrated that changes in cellular properties and synaptic connections underlie neurobehavioral plasticity during morphallaxis. Sensory-to-giant interneuron connections, undetectable prior to body injury, emerged within hours of segment amputation. The appearance of functional synaptic transmission was followed by interneuron activation, coupling of giant fiber spiking to motor outputs and overt segmental shortening. The onset of morphallactic plasticity varied along the body axis and emerged more rapidly in segments closer to regions of sensory field overlap between the two giant fiber pathways. The medial and lateral giant fibers were simultaneously activated during a transient phase of network remodeling. Thus, synaptic plasticity at sensory-to-giant interneuron connections mediates escape circuit morphallaxis in this regenerating annelid worm. Copyright © 2011 Wiley Periodicals, Inc.

  13. Metabolic effects of glyphosate change the capacity of maize culture to regenerate plants.

    Science.gov (United States)

    Ulanov, Alexander; Lygin, Anatoliy; Duncan, David; Widholm, Jack; Lozovaya, Vera

    2009-06-01

    Since the presence of glyphosate in maize tissue cultures of proprietary line L2 was very detrimental to plant regeneration, we determined metabolic changes associated with the glyphosate effects on plant regeneration in maize cultures. The polar fraction composition and soluble and cell-wall-bound phenolics were analyzed in the regenerable (R) and non-regenerable (NR) calluses of maize line L2. The tissues with high regeneration capacity had low sugar and 4-aminobutyric acid (GABA) concentrations and increased concentrations of most amino acids, polyamines and indole-3-butenol in the soluble polar fraction and higher ferulic acid/coumaric acid and ferulic acid/diferulic acid ratios and higher levels of the predominant G (guaiacyl) units in the cell wall fraction compared with NR calluses as was found before with H99 and HiII maize R and NR tissues, indicating an association of these metabolites with the capacity of maize cultured tissue to regenerate plants. We also found that di-coumaroyl spermidine and coumaroyl-feruoyl spermidine are present in the soluble fraction of L2 R tissues and are practically absent in NR tissues. However, we did not see such differences in HiII and H99 samples, which indicate that these are genotypic features not related to the capacity to regenerate plants in maize tissue cultures. Glyphosate treatment caused the accumulation of shikimic and quinic acids (not detected in untreated samples) in R and NR calluses (with higher levels found in R tissues) and also decreased the FA/diFA ratio in cell wall phenolics, polyamine and amino acid levels, and increased sugar concentrations in the R L2 tissues, indicating a metabolic shift of R callus to NR tissues.

  14. In vitro plant regeneration from Narbon Vetch (Vicia narbonensis L ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-07-18

    Jul 18, 2008 ... photoperiod. The pH of all cultures was adjusted to 5.6 - 5.8 before adding 0.65% agar (Duchefa) and autoclaving at 121oC, 118 kPa pressure for 20 min. To induce roots, the shoots regenerated on all .... geographic origins for very successful tissue culture in the future. Moreover, it is expected that the ...

  15. Callus production and regeneration of the medicinal plant Papaver ...

    African Journals Online (AJOL)

    Results showed that the best media for callus induction of P. orientale consisted of the MS basic media supplemented with 0.5 mg/l BA and 0.5 or 1 mg/l NAA. Shoots were regenerated in cultures grown on MS medium containing 0.5 mg/l Kin and (0.5 or 1.0 mg/l) NAA and were induced to root on B5 medium containing 1 ...

  16. Somatic embryogenesis and plant regeneration in Wedelia calendulacea Less. an endangered medicinal plant

    Directory of Open Access Journals (Sweden)

    Shamima Akhtar Sharmin

    2014-06-01

    Full Text Available In this work, plant regeneration via somatic embryogenesis was achieved from leaf and internode derived callus of Wedelia calendulacea, an endangered medicinal plant. Primary callus was induced by culturing leaf disc and internode explant on Murashige and Skoog medium supplemented with 2.0 mg L-1 of 2,4-D under light condition. Transfer of embryogenic callus on a reduced concentration of 2,4-D facilitated somatic embryo development while calluses remained unorganized at the same 2,4-D level. A histological analysis confirmed somatic embryo by revealing the presence of a closed vascular system in the developing embryos and lack of a vascularconnection with surrounding callus tissues. Somatic embryos germinated into plantlets upon transfer on MS medium containing 1.0 mg L-1 BAP plus 0.5 mg L-1 GA3. Plantlets were acclimatized successfully and survived under soil condition. This is the first on somatic embryogenesis of W.calendulacea. This result could facilitate genetic transformation of this important medicinal plant.

  17. Efficient culture protocol for plant regeneration from cotyledonary petiole explants of Jatropha curcas L.

    Directory of Open Access Journals (Sweden)

    Ying Liu

    2016-09-01

    Full Text Available A high-frequency and reproducible protocol for induction of adventitious shoot buds and plant regeneration from cotyledonary petiole explants of Jatropha curcas L. has been developed. The cotyledonary petiole explants of J. curcas cultured directly on medium supplemented with thidiazuron (TDZ induce regeneration of poor quality shoot buds that have a low regeneration frequency. However, treating the explants with high concentrations (10–60 mg/L of TDZ solution for certain time periods (5–80 min significantly increased the regeneration frequency and improved the quality of the regenerated shoot buds. The best shoot buds induction (88.42% and number of shoot buds (12.67 per explant were observed when in vitro explants were treated with 20 mg/L TDZ solution for 20 min before being transferred on hormone-free medium after 30 days. Regeneration was also influenced by the orientation (horizontal or vertical of the explants on the medium, and by the origin of the cotyledonary petioles (in vitro or in vivo used for the preparation of explants. We performed subsequent experiments for elongation and rooting of the regenerated shoot buds. Addition of L-arginine to the medium was conducive to the elongation of the shoot buds. A concentration of 7.5 mg/L L-arginine yielded the best results. The elongated shoots could initiate roots to become intact plantlets in half-strength Murashige and Skoog medium containing 0.1 mg/L indole-3-butyric acid. After acclimatization, these plantlets could be transplanted to the soil and the growth was normal. Therefore, application of the methods described here helped to increase plant regeneration efficiency.

  18. Initial response of individual soft mast-producing plants to different forest regeneration methods in the Ouachita Mountains

    Science.gov (United States)

    Roger W. Perry; Ronald E. Thill; Philip A. Tappe; David G. Peitz

    2004-01-01

    Abstract - Recent policy changes have eliminated clearcutting as the primary pine regeneration method on Federal lands in the Southern United States. However, the effects of alternative natural regeneration methods on soft mast production are unknown. We compared plant coverage and mast production of 37 soft mast-producing plants among four...

  19. Regeneration of Dioscorea floribunda plants from cryopreserved encapsulated shoot tips: effect of plant growth regulators.

    Science.gov (United States)

    Mandal, B B; Ahuja-Ghosh, Sangeeta

    2007-01-01

    The encapsulation-dehydration protocol for the cryopreservation of in vitro shoot tips of Dioscorea floribunda was optimized. Maximum survival of 87% was obtained when overnight pretreatment with 0.3 M sucrose was followed by encapsulation, preculture in 0.75 M sucrose for 4 d, dehydration in a laminar air flow for 5.5 h, quenching in liquid nitrogen and thawing at 40 degrees C. During recovery growth, 29% shoot formation was obtained when cryopreserved shoot tips were initially cultured for 25 d on a medium with 1.5 mg per liter (-1) BAP, 0.2 mg per liter(-1) NAA and 0.2 mg per liter(-1) GA3 followed by culturing for 15 d on a medium with reduced BAP (1 mg per liter(-1)) but increased NAA (0.5 mg per liter(-1)) and GA3 (0.3 mg per liter(-1)). Finally, transfer on to a medium with further reduced doses of BAP (0.05 mg per liter(-1)) and NAA (0.15 mg per liter(-1)) but without GA3 stimulated production of fully grown plantlets. All plants regenerated without callus formation. Modification of post-thaw culture media with plant growth regulators was essential for regrowth of shoot tips to plantlets.

  20. Callus induction and plant regeneration in vitro in Actinidia

    Directory of Open Access Journals (Sweden)

    Adam Matkowski

    2014-01-01

    Full Text Available The dependence of callus induction and organogenesis on auxins (2,4-D, IAA and cytokinins (BAP, KIN content in MS medium in Actinidia arguta, A. deliciosa and A. polygama and explants derived from stems, petioles, leaves and roots was investigated. The ability to form callus was good for stem and petiole explants and weaker for leaf and root explants in all three species. The media with low level of 2,4-D (0.01 and 0.1 mg/l and higher of cytokinins (1.0 and 5.0 mg/l promoted callus growth and viability. Higher 2,4-D as well as IAA concentrations caused degeneration of calli. The application of BAP was more advantageous than of KIN in the case of A. arguta and A. polygama. Organogenesis was quick and abundant after passage to media with 0.1-5.0 mg/l of IAA and 1.0-5.0 mg/l of cytokinins only in callus derived from stems. Calli from the other explants showed weaker ability to regenerate. The reactions of the three investigated species, differed slightly, the similarities were however, more significant. Shoots regenerated from callus were rooted and the plantlets were transferred to soil.

  1. High capacity of plant regeneration from callus of interspecific hybrids with cultivated barley (Hordeum vulgare L.)

    DEFF Research Database (Denmark)

    Bagger Jørgensen, Rikke; Jensen, C. J.; Andersen, B.

    1986-01-01

    Callus was induced from hybrids between cultivated barley (Hordeum vulgare L. ssp. vulgare) and ten species of wild barley (Hordeum L.) as well as from one backcross line ((H. lechleri .times. H. vulgare) .times. H. vulgare). Successful callus induction and regeneration of plants were achieved fr...

  2. Somatic embryogenesis and plant regeneration of northern red oak (Quercus rubra L.)

    Science.gov (United States)

    G. Vengadesan; Paula M. Pijut

    2009-01-01

    A somatic embryogenesis protocol for plant regeneration of northern red oak (Quercus rubra) was established from immature cotyledon explants. Embryogenic callus cultures were induced on Murashige and Skoog medium (MS) containing 3% sucrose, 0.24% Phytagel™, and various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) after 4 weeks of...

  3. Effect of Salicylic Acid on Somatic Embryogenesis and Plant Regeneration in Hedychium bousigonianum

    Science.gov (United States)

    The objective of this study was to induce somatic embryogenesis in Hedychium bousigonianum Pierre ex Gagnepain and assess the influence of salicylic acid (S) on somatic embryogenesis. Somatic embryos and subsequently regenerated plants were successfully obtained 30 days after transfer of embryogenic...

  4. Putting community data to work: some understory plants indicate red spruce regeneration habitat

    Science.gov (United States)

    Alison C. Dibble; John C. Brissette; Malcolm L. Hunter

    1999-01-01

    When harvested, red spruce (Picea rubens) at low elevations is vulnerable to temporary displacement by balsam fir (Abies balsamea) and hardwoods. If indicator plants can be found by which to assess spruce regeneration habitat, then biota dependent on red spruce dominance could benefit. Associations between spruce seedlings (0.1-0.5...

  5. In vitro culture and plant regeneration derived from ray florets of ...

    African Journals Online (AJOL)

    Nine cultivars of Chrysanthemum morifolium were screened using the ray floret explants to determine the capability for plant regeneration on four media protocols and subsequently to find out the best genotype source linked with the optimum medium conditions for the high potentiality of shoot formation. The results ...

  6. Isolation of protoplasts and culture and regeneration into plants in Alstroemeria

    NARCIS (Netherlands)

    Kim, J.B.; Bergervoet-van Deelen, J.E.M.; Raemakers, C.J.J.M.; Jacobsen, E.; Visser, R.G.F.

    2005-01-01

    An efficient system for the regeneration of plants from protoplasts was developed in Alstroemeria. Friable embryogenic callus (FEC) proved to be the best source for protoplast isolation and culture when compared with leaf tissue and compact embryogenic callus. Protoplast isolation was most efficient

  7. Development of long-term and reliable in vitro plant regeneration ...

    African Journals Online (AJOL)

    The response to in vitro tissue culture of five important Mexican malting barley cultivars namely 'Armida', 'Esmeralda', 'Adabella', 'Esperanza' and 'Alina', was evaluated. Callus induction and plant regeneration were evaluated in shoot apices and immature barley embryos harvested eight, 12, 16, 20 and 24 days after ...

  8. In vitro plant regeneration of Esmeralda clarkei Rchb.f. via protocorm ...

    African Journals Online (AJOL)

    user

    2012-07-05

    Jul 5, 2012 ... Esmeralda clarkei is an epiphytic native orchid species of Nepal growing under medium amount of light with fragrant flowers. Reliable protocols for in vitro plant regeneration of E. clarkei via protocorm explants were developed. Protocorms obtained from in vitro germinated seeds cultured on Murashige.

  9. Plant regeneration and ploidy level stability in Carica papaya ...

    African Journals Online (AJOL)

    Usuario

    2013-04-11

    Apr 11, 2013 ... Comparatively, hermaphrodite plants exhibit considerable variation in regards to yield, fruit quality, and ..... Renukdas N, Mohan ML, Khuspe SS, Rawal SK (2003). Influence of boron on somatic embryogenesis in papaya. Biol. Plant. 47: 129-132. Reuveni O, Shlesinger DR, Lavi U (1990). In vitro clonal ...

  10. In vitro regeneration of a common medicinal plant, Ocimum sanctum ...

    African Journals Online (AJOL)

    aristo_team

    for further research and germplasm preservation. In this paper, our point is to portray a straightforward and solid convention to increase this pharmaceutically important plant through high-recurrence axillary shoot multiplication. MATERIALS AND METHODS. O. sanctum plants, gathered from an habitation of Hindu religious.

  11. Plant regeneration of Lotononis bainesii Baker (Fabaceae) through ...

    African Journals Online (AJOL)

    María Laura Vidoz

    2012-05-22

    May 22, 2012 ... Crawley, WA 6009, Australia. 4School of Plant Biology, Faculty of Natural and Agricultural Sciences, The University of Western Australia, 35 Stirling. Highway, Crawley, WA 6009, Australia. 5Plant Molecular and Cellular Biology Program, Genetics Institute, University of Florida, Gainesville, FL 32611-0300,.

  12. Regeneration of Centella asiatica plants from non-embryogenic cell lines and evaluation of antibacterial and antifungal properties of regenerated calli and plants

    Science.gov (United States)

    2011-01-01

    Background The threatened plant Centella asiatica L. is traditionallyused for a number of remedies. In vitro plant propagation and enhanced metabolite production of active metabolites through biotechnological approaches has gained attention in recent years. Results Present study reveals that 6-benzyladenine (BA) either alone or in combination with 1-naphthalene acetic acid (NAA) supplemented in Murashige and Skoog (MS) medium at different concentrations produced good quality callus from leaf explants of C. asiatica. The calli produced on different plant growth regulators at different concentrations were mostly embryogenic and green. Highest shoot regeneration efficiency; 10 shoots per callus explant, from non-embryogenic callus was observed on 4.42 μM BA with 5.37 μM NAA. Best rooting response was observed at 5.37 and 10.74 μM NAA with 20 average number of roots per explant. Calli and regenerated plants extracts inhibited bacterial growth with mean zone of inhibition 9-13 mm diameter when tested against six bacterial strains using agar well diffusion method. Agar tube dilution method for antifungal assay showed 3.2-76% growth inhibition of Mucor species, Aspergillus fumigatus and Fusarium moliniformes. Conclusions The present investigation reveals that non-embryogenic callus can be turned into embryos and plantlets if cultured on appropriate medium. Furthermore, callus from leaf explant of C. asiatica can be a good source for production of antimicrobial compounds through bioreactor. PMID:21989222

  13. Regeneration of Centella asiatica plants from non-embryogenic cell lines and evaluation of antibacterial and antifungal properties of regenerated calli and plants

    Directory of Open Access Journals (Sweden)

    Habib Darima

    2011-10-01

    Full Text Available Abstract Background The threatened plant Centella asiatica L. is traditionallyused for a number of remedies. In vitro plant propagation and enhanced metabolite production of active metabolites through biotechnological approaches has gained attention in recent years. Results Present study reveals that 6-benzyladenine (BA either alone or in combination with 1-naphthalene acetic acid (NAA supplemented in Murashige and Skoog (MS medium at different concentrations produced good quality callus from leaf explants of C. asiatica. The calli produced on different plant growth regulators at different concentrations were mostly embryogenic and green. Highest shoot regeneration efficiency; 10 shoots per callus explant, from non-embryogenic callus was observed on 4.42 μM BA with 5.37 μM NAA. Best rooting response was observed at 5.37 and 10.74 μM NAA with 20 average number of roots per explant. Calli and regenerated plants extracts inhibited bacterial growth with mean zone of inhibition 9-13 mm diameter when tested against six bacterial strains using agar well diffusion method. Agar tube dilution method for antifungal assay showed 3.2-76% growth inhibition of Mucor species, Aspergillus fumigatus and Fusarium moliniformes. Conclusions The present investigation reveals that non-embryogenic callus can be turned into embryos and plantlets if cultured on appropriate medium. Furthermore, callus from leaf explant of C. asiatica can be a good source for production of antimicrobial compounds through bioreactor.

  14. Rapid identification of the medicinal plant Taraxacum formosanum ...

    African Journals Online (AJOL)

    Rapid identification of the medicinal plant Taraxacum formosanum and distinguishing of this plant from its adulterants by ribosomal DNA internal transcribed spacer (ITS) based DNA barcode. YC Chiang, WT Chang, MD Chen, GH Lai, HJ Chen, J Chao, MK Lin, YS Chang, YM Chou, MS Lee, MS Lee ...

  15. Thidiazuron-induced high frequency of shoot induction and plant regeneration in protoplast derived pea callus.

    Science.gov (United States)

    Böhmer, P; Meyer, B; Jacobsen, H J

    1995-01-01

    Protoplasts isolated from lateral shoot buds of cotyledon-free pea embryo axes were regenerated to callus. Protoplast derived calluses with a diameter of about 1cm were transferred to shoot induction media, containing different concentrations (1-50µM) of thidiazuron. Shoot formation was observed after 16 weeks up to 12% efficiency. Thidiazuron (10µM) was the most effective concentration in all experiments. Shoot buds elongated in medium supplemented with N-isopentenyl adenine and indole-3-butyric acid. Since rooting was almost impossible in these thidiazuron-induced shoots, shoots were grafted onto young pea seedlings and regenerated to fertile plants.

  16. Plant Regeneration and Somatic Embryogenesis from Immature Embryos Derived through Interspecific Hybridization among Different Carica Species

    Directory of Open Access Journals (Sweden)

    Latifah Amin

    2012-12-01

    Full Text Available Plant regeneration and somatic embryogenesis through interspecific hybridization among different Carica species were studied for the development of a papaya ringspot virus-resistant variety. The maximum fruit sets were recorded from the cross of the native variety C. papaya cv. Shahi with the wild species C. cauliflora. The highest hybrid embryos were recorded at 90 days after pollination and the embryos were aborted at 150 days after pollination. The immature hybrid embryos were used for plant regeneration and somatic embryogenesis. The 90-day-old hybrid embryos from the cross of C. papaya cv. Shahi × C. cauliflora showed the highest percentage of germination, as well as plant regeneration on growth regulators free culture medium after 7 days pre-incubation on half-strength MS medium supplemented with 0.2 mg/L BAP, 0.5 mg/L NAA and 60 g/L sucrose. The 90-day-old hybrid embryos from the cross of C. papaya cv. Shahi × C. cauliflora produced maximum callus, as well as somatic embryos when cultured on half-strength MS medium containing 5 mg/L 2,4-D, 100 mg/L glutamine, 100 mg/L casein hydrolysate and 60 g/L sucrose. The somatic embryos were transferred into half-strength MS medium containing 0.5 mg/L BAP and 0.2 mg/L NAA and 60 g/L sucrose for maturation. The highest number of regenerated plants per hybrid embryo (10.33 was recorded from the cross of C. papaya cv. Shahi × C. cauliflora. Isoenzyme and dendrogram cluster analysis using UPGMA of the regenerated F1 plantlets confirmed the presence of the hybrid plantlets.

  17. Plant regeneration and somatic embryogenesis from immature embryos derived through interspecific hybridization among different Carica species.

    Science.gov (United States)

    Azad, Md Abul Kalam; Rabbani, Md Golam; Amin, Latifah

    2012-12-12

    Plant regeneration and somatic embryogenesis through interspecific hybridization among different Carica species were studied for the development of a papaya ringspot virus-resistant variety. The maximum fruit sets were recorded from the cross of the native variety C. papaya cv. Shahi with the wild species C. cauliflora. The highest hybrid embryos were recorded at 90 days after pollination and the embryos were aborted at 150 days after pollination. The immature hybrid embryos were used for plant regeneration and somatic embryogenesis. The 90-day-old hybrid embryos from the cross of C. papaya cv. Shahi × C. cauliflora showed the highest percentage of germination, as well as plant regeneration on growth regulators free culture medium after 7 days pre-incubation on half-strength MS medium supplemented with 0.2 mg/L BAP, 0.5 mg/L NAA and 60 g/L sucrose. The 90-day-old hybrid embryos from the cross of C. papaya cv. Shahi × C. cauliflora produced maximum callus, as well as somatic embryos when cultured on half-strength MS medium containing 5 mg/L 2,4-D, 100 mg/L glutamine, 100 mg/L casein hydrolysate and 60 g/L sucrose. The somatic embryos were transferred into half-strength MS medium containing 0.5 mg/L BAP and 0.2 mg/L NAA and 60 g/L sucrose for maturation. The highest number of regenerated plants per hybrid embryo (10.33) was recorded from the cross of C. papaya cv. Shahi × C. cauliflora. Isoenzyme and dendrogram cluster analysis using UPGMA of the regenerated F(1) plantlets confirmed the presence of the hybrid plantlets.

  18. In vitro plant regeneration from different explants of Cardiospermum ...

    African Journals Online (AJOL)

    A rapid and efficient protocol was developed for inducing indirect organogenesis using stem and leaf explants of Cardiospermum halicacabum L. Explants were cultured on MS medium supplemented with different concentrations of IAA, NAA and 2, 4-D (1.0 to 2.0 mg/l) combined with 0.5 mg/l BAP for callus induction.

  19. Regeneration in bipinnaria larvae of the bat star Patiria miniata induces rapid and broad new gene expression

    Science.gov (United States)

    Oulhen, Nathalie; Heyland, Andreas; Carrier, Tyler J.; Zazueta-Novoa, Vanesa; Fresques, Tara; Laird, Jessica; Onorato, Thomas M.; Janies, Daniel; Wessel, Gary

    2016-01-01

    Background Some metazoa have the capacity to regenerate lost body parts. This phenomenon in adults has been classically described in echinoderms, especially in sea stars (Asteroidea). Sea star bipinnaria larvae can also rapidly and effectively regenerate a complete larva after surgical bisection. Understanding the capacity to reverse cell fates in the larva is important from both a developmental and biomedical perspective; yet, the mechanisms underlying regeneration in echinoderms are poorly understood. Results Here, we describe the process of bipinnaria regeneration after bisection in the bat star Patiria miniata. We tested transcriptional, translational, and cell proliferation activity after bisection in anterior and posterior bipinnaria halves as well as expression of SRAP, reported as a sea star regeneration associated protease (Vickery et al., 2001b). Moreover, we found several genes whose transcripts increased in abundance following bisection, including: vasa, dysferlin, vitellogenin 1 and vitellogenin 2. Conclusion These results show a transformation following bisection, especially in the anterior halves, of cell fate reassignment in all three germ layers, with clear and predictable changes. These results define molecular events that accompany the cell fate changes coincident to the regenerative response in echinoderm larvae. PMID:27555501

  20. [Transformation of sainfoin by Agrobacterium rhizogenes LBA9402 Bin19 and regeneration of transgenic plants].

    Science.gov (United States)

    Xu, Z Q; Ma, H J; Hao, J G; Jia, J F

    2000-03-01

    Hypocotyl segments of Onobrychis viciaefolia were transformed by Agrobacterium rhizogenes LBA9402 which harboured pBin19 and pRi1855. Seedling age and preculture time of hypocotyl segments influenced the transformation frequency. Paper electrophoresis revealed that 70% of single hairy root cultures could synthesize agropine. Calli were induced from hairy root segments on MS medium containing 0-9.05 mumol/L 2,4-D and 0-2.22 mumol/L 6-BA at first, then they were transferred onto MS0 medium without kanamycin for regeneration. Constitution and concentration of phytohormones in callus induction media affected subsequent regeneration of calluses on MS0 medium remarkably. Regeneration frequency and shoot number per callus declined when 2,4-D concentration in callus induction media increased from 4.52 to 9.05 mumol/L, while they ascended when 6-BA in callus induction media increased from 0 to 2.22 mumol/L. On MS medium supplemented with 4.52 mumol/L 2,4-D and 2.22 mumol/L 6-BA, only 14.2% hairy root segments could produce calluses, but the regeneration frequency reached 58.1% and the shoot number per callus was 37.2. In 32 analysed plants regenerated from 8 kanamycin resistant hairy root lines, 25 were nptII positive and showed different copy numbers.

  1. In vitro plant regeneration from leaves and internode sections of sweet cherry cultivars (Prunus avium L.).

    Science.gov (United States)

    Matt, Andrea; Jehle, Johannes A

    2005-10-01

    Regeneration of adventitious shoots from leaves and, for the first time, from internode sections were compared and optimized for five economically important sweet cherry cultivars, i.e. "Schneiders", "Sweetheart", "Starking Hardy Giant", "Kordia" and "Regina" (Prunus avium L.). The influence of basal media, carbon source, combination and dosage of phytohormones, ethylene inhibitor such as silver thiosulfate and a 16 h:8 h light:dark photoperiod versus complete darkness were evaluated. Both, DKW/WPM (1:1) and Quoirin/Lepoivre (QL) basal media stimulated organogenesis more than QL/WPM (1:1), Chee and Pool (CP), Murashige Skoog (MS), Driver and Kuniyuki (DKW) or woody plant (WPM) media did. An induction phase in darkness resulted in lower or zero regeneration rates. The best regeneration efficiencies were generally obtained with thidiazuron in combination with indole-3-butyric-acid. The addition of silver thiosulfate resulted in a similar or reduced regeneration efficiency. Significant genotypic variability in adventitious bud formation was evident for both explant sources, leaf and internode section. Adventitious shoots were obtained from 11% of leaf explants and 50% of internode sections indicating that shoot regeneration from internodes was significantly more efficient than from leaves.

  2. Plant Regeneration Through Tissue Culture Of Pear Millet ...

    African Journals Online (AJOL)

    Many problems were kipt out with immature embryos and shoot explants pieces used ; principally : Calli were initiated in plant flowering periode (every 110 days), and the low percentage of embrogenesis what either the source used. So for Pennisetum Glaucum, new embryogenic calli sources were explored from calli ...

  3. Plant regeneration from immature embryos of Kenyan maize inbred ...

    African Journals Online (AJOL)

    Field grown, self pollinated maize genotypes were planted in KARI (Kiboko and Kabete) research stations between January 2004 and May 2005. Immature maize embryos from twelve parental inbred lines and their respective single cross hybrids were evaluated for their ability form callus, somatic embryos and subsequent ...

  4. Plant regeneration from leaf-derived callus in Plectranthus barbatus ...

    African Journals Online (AJOL)

    A protocol was developed for Plectranthus barbatus for high frequency shoot organogenesis from leaf derived callus of aseptically grown plantlets derived from mature plant explants of high yielding clones (yield of forskolin 1.5 to 1.9%) on Gamborg medium (B5) medium supplemented with 2 mg/l 2,4- dichloro phenoxy ...

  5. Establishment of in vitro plant regeneration system for ...

    African Journals Online (AJOL)

    ajl2

    2012-06-05

    Jun 5, 2012 ... Two new tryptamine-derived alkaloids from Chimonanthus praecoxf concolor. Tetrahedron Lett., 47: 3199-3202. Kozomara B,Vinterhalter B, Radojević LJ, Vinterhalter D (2008). In vitropropagation of Chimonanthus praecox (L.), a winter flowering ornamental shrub. In Vitro Cell. Dev. Biol. Plant, 44: 142-147.

  6. An effective disinfection protocol for plant regeneration from shoot tip ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-06-03

    Jun 3, 2009 ... induced to form adventitious roots. The plantlets were maintained in the same medium and extra time of illumination so that their rooting systems may be deve- loped properly. The in vitro grown plantlets were then planted in 1.5 inches plastic trays with peat moss, vermiculite and perlite mixture (1:1:1 ratio) ...

  7. Plant regeneration of Brassica oleracea subsp. italica (Broccoli) CV ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-06-03

    Jun 3, 2009 ... Hypocotyl and shoot tip explants, 5-8 mm in size, were excised from 6-day-old broccoli seedlings. The explants were cultured on. MS medium incorporated with different concentrations of plant growth regulators for shoot proliferation and root formation. For shoot induction and multiplication from hypocotyl ...

  8. Callus production and regeneration of the medicinal plant Papaver ...

    African Journals Online (AJOL)

    Administrator

    2011-09-19

    Sep 19, 2011 ... Biologia, 65(4):. 647-652. Schuchmann R, Willmann E (1983). Somatic embryogenesis of tissues cultures of Papaver somniferium and Papaver orientale and its relationship to alkaloid and lipid metabolism. Plant Cell Rep. 2(2): 88-. 91. Shafiee A, Lalezari I, Nasseri-Nouri P, Asgharian R (1975). Alkaoids of.

  9. Direct multiple shoot induction and plant regeneration from dormant ...

    African Journals Online (AJOL)

    Administrator

    2011-09-07

    Sep 7, 2011 ... seedlings are raised the year before and transplanted in the next spring, which is labor intensive and time- consuming. Many successful examples for micropropagation and cryopreservation using overground dormant buds can be found in garden plants (Jacobsen and Dohmen 1990;. Mistretta et al., 1991).

  10. Regeneration of transgenic citrus plants under non selective conditions results in high-frequency recovery of plants with silenced transgenes.

    Science.gov (United States)

    Domínguez, A; Fagoaga, C; Navarro, L; Moreno, P; Peña, L

    2002-06-01

    Insertion of foreign DNA into plant genomes frequently results in the recovery of transgenic plants with silenced transgenes. To investigate to what extent regeneration under selective conditions limits the recovery of transgenic plants showing gene silencing in woody species, Mexican lime [ Citrus aurantifolia (Christm.) Swing.] plants were transformed with the p25 coat protein gene of Citrus tristeza virus (CTV) with or without selection for nptII and uidA. Strikingly, more than 30% of the transgenic limes regenerated under non-selective conditions had silenced transgenes, and in all cases silencing affected all the three transgenes incorporated. These results indicate that the frequency of transgene silencing may be greatly underestimated when the rate of silencing is estimated from the number of regenerants obtained under selective conditions. To our knowledge, this is the first report in which the frequency of gene silencing after transformation has been quantified. When the integration pattern of T-DNA was analyzed in silenced and non-silenced lines, it was observed that inverted repeats as well as direct repeats and even single integrations were able to trigger gene silencing. Gene silencing has often been associated with the insertion of DNA sequences as inverted repeats. Interestingly, here, direct repeats and single-copy insertions were found in both silenced and non-silenced lines, suggesting that the presence of inverted-repeat T-DNAs and the subsequent formation of dsRNAs triggering gene silencing cannot account for all silencing events.

  11. Plant regeneration from cell suspension-derived protoplasts of Saintpaulia ionantha Wendl.

    Science.gov (United States)

    Hoshino, Y; Nakano, M; Mii, M

    1995-03-01

    Friable calli were induced on leaf segments of Saintpaulia ionantha Wendl. on B5 medium containing 1 mg l(-1) 2,4-D and 2 g l(-1) casein hydrolysate. Cell suspension cultures were readily established from these friable calli and protoplasts could be isolated from the cells with yields of 1-3×10(7)/g f. wt.. By culturing in 0.1 % gellan gum-solidified B5 medium supplemented with 1 mg l(-1) 2,4-D and 0.1 M each of sucrose and mannitol at a density of 1×10(5)/ml, the protoplasts divided within 6 days and formed macro-colonies after 2 months of culture. Shoot regeneration from protoplast-derived calli was obtained by sequential treatment of the calli with plant growth regulators: initially with 1 mg l(-1) each of NAA and BA for 2 months followed by 0.01 mg l(-1) NAA and 5 mg l(-1) BA for 4 months. Regenerated plants were established after rooting of the shoots on half-strength MS medium, and successfully transferred to the greenhouse. The regenerated plants grew into flowering stage and showed the same phenotype as the parent plant.

  12. Histology, histochemistry and SEM are useful tools to study regeneration processes in plant tissue culture

    Directory of Open Access Journals (Sweden)

    Piotr Żabicki

    2013-04-01

    Full Text Available Tissue cultures in vitroare used for the multiplication of plants via direct and indirect (via callus regeneration. This approach is commonly applied in the protection of endangered species by the introduction of regenerated in vitro plantlets to botanical gardens and to the nature (so called ex situ plant conservation. In vitroconditions, especially the supplementation of tissue culture media with plant growth regulators, cause a somaclonal variation, resulting in genetic differences among regenerated plants. To analyze callus structure, including cell shapes and sizes, cell differentiation (e.g. the presence of xylem vessels and regeneration processes (organogenesis, somatic embryogenesis, the histological, histochemical and SEM techniques are applied. In this study, to obtain regeneration of plants in culture conditions, we have used three Viola species (V. epipsilaLedeb., V. stagnina Kit. and V. uliginosaBesser, indicated to be critically endangered according to Polish Red Book of Plants (Kazmierczakowa & Zarzycki 2001 and two genotypes of a model plant Arabidopsis thaliana(L. Heynh. (Columbia-0 and an insertional cdkg ;2mutant line. An Arabidopsis homozygous cdkg ;2 knock-out originated from a T 3 generation of T-DNA insertional line SALK_090262 (Alonso et al. 2003 and has been selected from a subsequent T 4 generation based on PCR analysis using primers complementary to flanking positions of full-length cDNA of CDKG;2gene product (a clone isolated by Seki et al. 2002. The aims of the study were: 1 to select the most convenient method to obtain regenerated Violaplants with maternal genotype i.e., via direct organogenesis or somatic embryogenesis; 2 to determine the effect of mutation in CDKG;2 gene on the explant response to in vitroconditions, including callus proliferation and regeneration. In three Viola species organogenesis was induced on MS (Murashige and Skoog basal medium supplied with thidiazuron (TDZ in concentrations 0.5 mg

  13. Efficient plant regeneration from leaf explants of Solanum americanum

    African Journals Online (AJOL)

    ONOS

    2010-09-06

    Sep 6, 2010 ... (Solanum dulcamara L.), a medicinal plant. Acta Soc. Bot. Pol. 77(4):. 275-280. Patiño-Torres C, Hoyos-Sánchez R, Afanador-Kafuri L (2007). Selección y regeneración in vitro de somaclones de tomate de árbol. (Solanum betacea cav. Sendt) utilizando filtrados de cultivo de. Colletotrichum acutatum con ...

  14. Slicing across Kingdoms: Regeneration in Plants and Animals

    OpenAIRE

    Birnbaum, Kenneth D; Alvarado, Alejandro Sánchez

    2008-01-01

    Multicellular organisms possessing relatively long life spans are subjected to diverse, constant, and often intense intrinsic and extrinsic challenges to their survival. Animal and plant tissues wear out as part of normal physiological functions and can be lost to predators, disease, and injury. Both kingdoms survive this wide variety of insults by strategies that include the maintenance of adult stem cells or the induction of stem cell potential in differentiated cells. Repatterning mechanis...

  15. Twelve-year responses of planted and naturally regenerating conifers to variable-retention harvest in the Pacific Northwest, USA

    Science.gov (United States)

    Lauren S. Urgenson; Charles B. Halpern; Paul D. Anderson

    2013-01-01

    We studied patterns of conifer regeneration over 12 years as part of a regional-scale experiment in variable-retention harvest in the Pacific Northwest, the DEMO Study. We compared survival and height growth of planted conifers and density and seral composition of natural regeneration among treatments with differing retention levels (15% versus 40%) and patterns (...

  16. The effects of three regeneration harvest methods on plant diversity and soil characteristics in the southern Appalachians

    Science.gov (United States)

    Katherine J. Elliott; Jennifer D. Knoepp

    2005-01-01

    We evaluated the effects of three regeneration harvest methods on plant diversity and soil resource availability in mixedhardwood ecosystems. The study area is in the Wine Spring Creek watershed on the Nantahala National Forest of the Southern Appalachian Mountains in western North Carolina. The regeneration treatments were: an irregular, two-aged shelterwood cut (2A...

  17. Regeneration of transgenic cassava plants (Manihot esculenta Crantz) from microbombarded embryogenic suspension cultures.

    Science.gov (United States)

    Schöpke, C; Taylor, N; Cárcamo, R; Konan, N K; Marmey, P; Henshaw, G G; Beachy, R N; Fauquet, C

    1996-06-01

    A protocol was established for the introduction of DNA into embryogenic suspension-derived tissues of cassava via microparticle bombardment, for the selection of genetically transformed cells, and for the regeneration of fully transgenic plants from these cells. The plasmid DNA used for bombardment contained a gene encoding neomycin phosphotransferase (nptII) and a gene encoding beta-glucuronidase (uidA). Selection of bombarded tissue with paromomycin resulted in the establishment of putative transgenic embryogenic calli. In most of these calli, beta-glucuronidase was detected histochemically. Molecular analysis of paromomycin-resistant embryogenic calli and of plants regenerated from these calli, confirmed the stable integration of bombarded DNA into the cassava genome.

  18. The caudal regeneration blastema is an accumulation of rapidly proliferating stem cells in the flatworm Macrostomum lignano.

    Science.gov (United States)

    Egger, Bernhard; Gschwentner, Robert; Hess, Michael W; Nimeth, Katharina T; Adamski, Zbigniew; Willems, Maxime; Rieger, Reinhard; Salvenmoser, Willi

    2009-07-15

    Macrostomum lignano is a small free-living flatworm capable of regenerating all body parts posterior of the pharynx and anterior to the brain. We quantified the cellular composition of the caudal-most body region, the tail plate, and investigated regeneration of the tail plate in vivo and in semithin sections labeled with bromodeoxyuridine, a marker for stem cells (neoblasts) in S-phase. The tail plate accomodates the male genital apparatus and consists of about 3,100 cells, about half of which are epidermal cells. A distinct regeneration blastema, characterized by a local accumulation of rapidly proliferating neoblasts and consisting of about 420 cells (excluding epidermal cells), was formed 24 hours after amputation. Differentiated cells in the blastema were observed two days after amputation (with about 920 blastema cells), while the male genital apparatus required four to five days for full differentiation. At all time points, mitoses were found within the blastema. At the place of organ differentiation, neoblasts did not replicate or divide. After three days, the blastema was made of about 1420 cells and gradually transformed into organ primordia, while the proliferation rate decreased. The cell number of the tail plate, including about 960 epidermal cells, was restored to 75% at this time point. Regeneration after artificial amputation of the tail plate of adult specimens of Macrostomum lignano involves wound healing and the formation of a regeneration blastema. Neoblasts undergo extensive proliferation within the blastema. Proliferation patterns of S-phase neoblasts indicate that neoblasts are either determined to follow a specific cell fate not before, but after going through S-phase, or that they can be redetermined after S-phase. In pulse-chase experiments, dispersed distribution of label suggests that S-phase labeled progenitor cells of the male genital apparatus undergo further proliferation before differentiation, in contrast to progenitor cells of

  19. Somatic embryogenesis and plant regeneration in Quercus acutissima.

    Science.gov (United States)

    Kim, Y W; Lee, B C; Lee, S K; Jang, S S

    1994-03-01

    Immature embryos of Quercus acutissima were collected weekly beginning 5 weeks post-fertilization and cultured on modified MS(Murashige and Skoog) medium containing 1,000 mg/l glutamine and 5 mM proline with different combinations of IBA(0.5-10.0 mg/l) and BA(0 or 1.0 mg/l) in light. The highest percentage of embryogenic cultures occurred on the medium containing 0.5 mg/l IBA or 1.0 mg/l BA and 0.5 mg/l IBA. Four weeks after initiation, the embryogenic cultures were transferred to MS medium without plant growth regulators and cultured for 4 weeks. The somatic embryos were then transferred to germination medium. The best germination results were achieved from WPM(Woody Plant Medium) containing 0.1 mg/l BA. Plantlets from somatic embryos were incubated on WPM supplemented with 0.2 mg/l BA for 4 weeks and plantlets with well developed shoots and roots were transplanted to perlite and peat moss(1∶1, v/v) mixtures and placed in a culture room. After being hardened off for 8 weeks, they were transferred outdoors where they grew.

  20. Plant Regeneration and Somatic Embryogenesis from Immature Embryos Derived through Interspecific Hybridization among Different Carica Species

    OpenAIRE

    Azad, Md. Abul; Rabbani, Md. Golam; Amin, Latifah

    2012-01-01

    Plant regeneration and somatic embryogenesis through interspecific hybridization among different Carica species were studied for the development of a papaya ringspot virus-resistant variety. The maximum fruit sets were recorded from the cross of the native variety C. papaya cv. Shahi with the wild species C. cauliflora. The highest hybrid embryos were recorded at 90 days after pollination and the embryos were aborted at 150 days after pollination. The immature hybrid embryos were used for pla...

  1. Fact Sheet - Final Air Toxics Rule for Steel Pickling and HCI Process Facilities and Hydrochloric Acid Regeneration Plants

    Science.gov (United States)

    Fact Sheet summarizing the main points of the national emssions standard for hazaradous air pollutants (NESHAP) for Steel Pickling— HCl Process Facilities and Hydrochloric Acid Regeneration Plants as promulgated on June 22, 1999.

  2. Loss of CMD2-mediated resistance to cassava mosaic disease in plants regenerated through somatic embryogenesis.

    Science.gov (United States)

    Beyene, Getu; Chauhan, Raj Deepika; Wagaba, Henry; Moll, Theodore; Alicai, Titus; Miano, Douglas; Carrington, James C; Taylor, Nigel J

    2016-09-01

    Cassava mosaic disease (CMD) and cassava brown streak disease (CBSD) are the two most important viral diseases affecting cassava production in Africa. Three sources of resistance are employed to combat CMD: polygenic recessive resistance, termed CMD1, the dominant monogenic type, named CMD2, and the recently characterized CMD3. The farmer-preferred cultivar TME 204 carries inherent resistance to CMD mediated by CMD2, but is highly susceptible to CBSD. Selected plants of TME 204 produced for RNA interference (RNAi)-mediated resistance to CBSD were regenerated via somatic embryogenesis and tested in confined field trials in East Africa. Although micropropagated, wild-type TME 204 plants exhibited the expected levels of resistance, all plants regenerated via somatic embryogenesis were found to be highly susceptible to CMD. Glasshouse studies using infectious clones of East African cassava mosaic virus conclusively demonstrated that the process of somatic embryogenesis used to regenerate cassava caused the resulting plants to become susceptible to CMD. This phenomenon could be replicated in the two additional CMD2-type varieties TME 3 and TME 7, but the CMD1-type cultivar TMS 30572 and the CMD3-type cultivar TMS 98/0505 maintained resistance to CMD after passage through somatic embryogenesis. Data are presented to define the specific tissue culture step at which the loss of CMD resistance occurs and to show that the loss of CMD2-mediated resistance is maintained across vegetative generations. These findings reveal new aspects of the widely used technique of somatic embryogenesis, and the stability of field-level resistance in CMD2-type cultivars presently grown by farmers in East Africa, where CMD pressure is high. © 2015 The Authors Molecular Plant Pathology Published by British Society for Plant Pathology and John Wiley & Sons Ltd.

  3. [Establishment of in vitro culture, plant regeneration and genetic transformation of Camelina sativa].

    Science.gov (United States)

    Emets, A I; Boĭchuk, Iu N; Shisha, E N; Rakhmetov, D B; Blium, Ia B

    2013-01-01

    The results on in vitro culture establishment, plantlet regeneration and rooting of Camelina sativa cultivar sample Peremozhets and cultivar Mirazh are presented. Effective concentrations of sterilizing agents and duration of plant material treatment were estimated. Phytohormone ratio, sucrose concentration in nutrient medium that induce effective formation of C. sativa shoots and NAA concentration for plantlet rooting have been established. The method of Agrobacterium-mediated transformation of Camelina by using binary vector pGH217 carrying reporter beta-glucoronidase (gus) gene driven under 35S CaMV promoter and nos-terminator, and selective marker hpt gene conferring hygromycin-resistance in transgenic plant was elaborated.

  4. Differences in vole preference, secondary chemistry and nutrient levels between naturally regenerated and planted Norway spruce seedlings.

    Science.gov (United States)

    Virjamo, Virpi; Julkunen-Tiitto, Riitta; Henttonen, Heikki; Hiltunen, Eveliina; Karjalainen, Reijo; Korhonen, Juhani; Huitu, Otso

    2013-10-01

    Field voles (Microtus agrestis) cause severe damage to young Norway spruce (Picea abies) plantations during wintertime in Fennoscandia. We experimentally investigated vole preference for winter-dormant, naturally regenerated seedlings; spring-planted seedlings; or autumn-planted seedlings; and how preference corresponds with seedling chemistry. Voles showed the highest preference for autumn-planted seedlings and the second highest for spring-planted seedlings, while naturally regenerated seedlings were avoided. The stems of the autumn-planted seedlings contained higher concentrations of nitrogen and piperidine alkaloids and lower concentrations of stilbenes than did the other groups. In addition to differences between naturally regenerated and planted seedlings, we investigated seasonal differences in naturally regenerated P. abies needle and bark secondary chemistry. While piperidine alkaloid concentrations did not vary with season, the soluble non-tannin phenolics of needles and the condensed tannins of bark were lower in May than in November or January. At the time of planting, the concentration of bark piperidine alkaloids was higher in autumn-planted than in spring-planted seedlings. We detected two alkaloids not previously found in P. abies, 2-methyl-6-propyl-1,6-piperideine and a tentatively identified pinidine-isomer. Our results demonstrate that vole choice of spruce seedlings is promoted by high nitrogen and low stilbene content, both associated with seedlings planted late in the season. As vole damage is linked to seedling chemistry, damage potentially could be mitigated by advancing planting or by manipulating plant chemistry in nurseries.

  5. Transformation of pecan and regeneration of transgenic plants.

    Science.gov (United States)

    McGranahan, G H; Leslie, C A; Dandekar, A M; Uratsu, S L; Yates, I E

    1993-09-01

    A gene transfer system developed for walnut (Juglans regia L.) was successfully applied to pecan (Carya illinoensis [Wang] K. Koch). Repetitively embryogenic somatic embryos derived from open-pollinated seed of 'Elliott', 'Wichita', and 'Schley' were co-cultivated with Agrobacterium strain EHA 101/pCGN 7001, which contains marker genes for beta-glucuronidase activity and resistance to kanamycin. Several modifications of the standard walnut transformation techniques were tested, including a lower concentration of kanamycin and a modified induction medium, but these treatments had no measurable effect on efficiency of transformation. Nineteen of the 764 viable inoculated embryos produced transgenic subclones; 13 of these were from the line 'Elliott'6, 3 from 'Schley'5/3, and 3 from 'Wichita'9. Transgenic embryos of 'Wichita'9 germinated most readily and three subclones were successfully micropropagated. Three transgenic plants of one of these subclones were obtained by grafting the tissue cultured shoots to seedling pecan rootstock in the greenhouse. Gene insertion, initially detected by GUS activity, was confirmed by detection of integrated T-DNA sequences using Southern analysis.

  6. Carbon dioxide absorber and regeneration assemblies useful for power plant flue gas

    Science.gov (United States)

    Vimalchand, Pannalal; Liu, Guohai; Peng, Wan Wang

    2012-11-06

    Disclosed are apparatus and method to treat large amounts of flue gas from a pulverized coal combustion power plant. The flue gas is contacted with solid sorbents to selectively absorb CO.sub.2, which is then released as a nearly pure CO.sub.2 gas stream upon regeneration at higher temperature. The method is capable of handling the necessary sorbent circulation rates of tens of millions of lbs/hr to separate CO.sub.2 from a power plant's flue gas stream. Because pressurizing large amounts of flue gas is cost prohibitive, the method of this invention minimizes the overall pressure drop in the absorption section to less than 25 inches of water column. The internal circulation of sorbent within the absorber assembly in the proposed method not only minimizes temperature increases in the absorber to less than 25.degree. F., but also increases the CO.sub.2 concentration in the sorbent to near saturation levels. Saturating the sorbent with CO.sub.2 in the absorber section minimizes the heat energy needed for sorbent regeneration. The commercial embodiments of the proposed method can be optimized for sorbents with slower or faster absorption kinetics, low or high heat release rates, low or high saturation capacities and slower or faster regeneration kinetics.

  7. Organogenesis and plant regeneration of Arachis villosa Benth. (Leguminosae) through leaf culture.

    Science.gov (United States)

    Fontana, María Laura; Mroginski, Luis Amado; Rey, Hebe Yolanda

    2009-12-01

    With the aim of developing an efficient plant regeneration protocol, leaflet explants of three accessions of Arachis villosa Benth. (S2866, S2867 and L97) were cultured on basic Murashige and Skoog medium supplemented with different combinations of plant growth regulators: alpha-naphthalenacetic acid, indole-3-butyric acid, 6-benzylaminopurine, kinetin and thidiazuron. The accession L97 was the only one able to differentiate buds through indirect organogenesis. The most suitable combination for bud regeneration was the basic medium added with 13.62 microM thidiazuron and 4.44 microM 6-benzylaminopurine. These results show the important role of the genotype in morphogenetic responses and the organogenetic effect of thidiazuron in Arachis villosa accession L97. A thidiazuron lacking media (only 0.54 microM alpha-naphthalenacetic acid, 13.95 microM kinetin and 13.32 microM 6-benzylaminopurine were added) promoted the elongation of the regenerated buds. Adventitious rooting was achieved 90 days after the isolated shoots were transferred to a rooting medium containing 0.54 microM alpha-naphthalenacetic acid.

  8. Plants survive rapid decompression: Implications for bioregenerative life support

    Science.gov (United States)

    Wheeler, R. M.; Wehkamp, C. A.; Stasiak, M. A.; Dixon, M. A.; Rygalov, V. Y.

    2011-05-01

    Radish (Raphanus sativus), lettuce (Latuca sativa), and wheat (Triticum aestivum) plants were grown at either 98 kPa (ambient) or 33 kPa atmospheric pressure with constant 21 kPa oxygen and 0.12 kPa carbon dioxide in atmospherically closed pressure chambers. All plants were grown rockwool using recirculating hydroponics with a complete nutrient solution. At 20 days after planting, chamber pressures were pumped down as rapidly as possible, reaching 5 kPa after about 5 min and ˜1.5 kPa after about 10 min. The plants were held at 1.5 kPa for 30 min and then pressures were restored to their original settings. Temperature (22 °C) and humidity (65% RH) controls were engaged throughout the depressurization, although temperatures dropped to near 16 °C for a brief period. CO2 and O2 were not detectable at the low pressure, suggesting that most of the 1.5 kPa atmosphere consisted of water vapor. Following re-pressurization, plants were grown for another 7 days at the original pressures and then harvested. The lettuce, radish, and wheat plants showed no visible effects from the rapid decompression, and there were no differences in fresh or dry mass when compared to control plants maintained continuously at 33 or 98 kPa. But radish storage root fresh mass and lettuce head fresh and dry masses were less at 33 kPa compared to 98 kPa for both the controls and decompression treatment. The results suggest that plants are extremely resilient to rapid decompression, provided they do not freeze (from evaporative cooling) or desiccate. The water of the hydroponic system was below the boiling pressure during these tests and this may have protected the plants by preventing pressures from dropping below 1.5 kPa and maintaining humidity near 1.5 kPa. Further testing is needed to determine how long plants can withstand such low pressure, but the results suggest there are at least 30 min to respond to catastrophic pressure losses in a plant production chamber that might be used for life

  9. A simple and rapid method for determining transgenic cotton plants.

    Science.gov (United States)

    Zhang, Baohong; Wang, Hongmei; Liu, Fang; Wang, Qinglian

    2013-01-01

    Determining transgenic events is a critical step for obtaining transgenic plants as well as the later stage of application. Traditional methods, such as Northern blotting and qRT-PCR, for determining transgenic events either require radioactively labeled substrates, expensive instruments, or long-time commitments, which result in lab and time-consuming as well as expensive costs. These methods also require destroying the transgenic events. In this chapter, we present a simple and rapid method for determining transgenic cotton plants in both laboratory and field conditions. This method is based on the sensitivity of transgenic and non-transgenic plants to a specific chemical, such as antibiotics or herbicides. This method will facilitate the screening of transgenic events, save time, reduce cost, and speed up the application of transgenic technology on cotton breeding and production. More important, this is a nondestructive bioassay method; the transgenic plants can be transferred into greenhouse or field for the later study after the detection process.

  10. Rapid, Long-Distance Electrical and Calcium Signaling in Plants.

    Science.gov (United States)

    Choi, Won-Gyu; Hilleary, Richard; Swanson, Sarah J; Kim, Su-Hwa; Gilroy, Simon

    2016-04-29

    Plants integrate activities throughout their bodies using long-range signaling systems in which stimuli sensed by just a few cells are translated into mobile signals that can influence the activities in distant tissues. Such signaling can travel at speeds well in excess of millimeters per second and can trigger responses as diverse as changes in transcription and translation levels, posttranslational regulation, alterations in metabolite levels, and even wholesale reprogramming of development. In addition to the use of mobile small molecules and hormones, electrical signals have long been known to propagate throughout the plant. This electrical signaling network has now been linked to waves of Ca(2+) and reactive oxygen species that traverse the plant and trigger systemic responses. Analysis of cell type specificity in signal propagation has revealed the movement of systemic signals through specific cell types, suggesting that a rapid signaling network may be hardwired into the architecture of the plant.

  11. Shoot regeneration from cotyledonary leaf explants of jatropha curcas: A biodiesel plant

    KAUST Repository

    Kumar, Nitish Chandramohana

    2010-03-07

    A simple, high frequency, and reproducible method for plant regeneration through direct organogenesis from cotyledonary leaf explants of Jatropha curcas was developed using Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ) or 6-benzyl aminopurine (BAP). Medium containing TDZ has greater influence on regeneration as compared to BAP. The induced shoot buds were transferred to MS medium containing 10 lM kinetin (Kn), 4.5 lM BAP, and 5.5 lM a-naphthaleneacetic acid (NAA) for shoot proliferation. The proliferated shoots could be elongated on MS medium supplemented with different concentrations and combinations of BAP, indole-3-acetic acid (IAA), NAA, and indole-3-butyric acid (IBA). MS medium with 2.25 lM BAP and 8.5 lM IAA was found to be the best combination for shoot elongation. However, significant differences in plant regeneration and shoot elongation were observed among the genotypes studied. Rooting was achieved when the basal cut end of elongated shoots were dipped in half strength MS liquid medium containing dif- ferent concentrations and combinations of IBA, IAA, and NAA for 4 days, followed by transfer to growth regulators free half strength MS medium supplemented 0.25 mg l-1 activated charcoal. Elongated shoot treated with 15 lM IBA, 5.7 lM IAA, and 11 lM NAA resulted in highest percent rooting. The rooted plants could be established in soil with more than 90% survival rate. The method developed may be useful in improvement of J. curcas through genetic modification. © Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2010.

  12. Plant regeneration through somatic embryogenesis and genome size analysis of Coriandrum sativum L.

    Science.gov (United States)

    Ali, Muzamil; Mujib, A; Tonk, Dipti; Zafar, Nadia

    2017-01-01

    In the present study, an improved plant regeneration protocol via primary and secondary somatic embryogenesis was established in two Co-1 and Rajendra Swathi (RS) varieties of Coriandrum sativum L. Callus was induced from root explants on 2, 4-D (0.5-2.0 mg/l) supplemented MS. The addition of BA (0.2 mg/l) improved callus induction and proliferation response significantly. The maximum callus induction frequency was on 1.0 mg/l 2, 4-D and 0.2 mg/l BA added MS medium (77.5 % in Co-1 and 72.3 % in RS). The callus transformed into embryogenic callus on 2, 4-D added MS with maximum embryogenic frequency was on 1.0 mg/l. The granular embryogenic callus differentiated into globular embryos on induction medium, which later progressed to heart-, torpedo- and cotyledonary embryos on medium amended with 0.5 mg/l NAA and 0.2 mg/l BA. On an average, 2-3 secondary somatic embryos (SEs) were developed on mature primary SEs, which increased the total embryo numbers in culture. Histology and scanning electron microscopy (SEM) studies are presented for the origin, development of primary and secondary embryos in coriander. Later, these induced embryos converted into plantlets on 1.0 mg/l BA and 0.2 mg/l NAA-amended medium. The regenerated plantlets were cultured on 0.5 mg/l IBA added ½ MS for promotion of roots. The well-rooted plantlets were acclimatized and transferred to soil. The genetic stability of embryo-regenerated plant was analyzed by flow cytometry with optimized Pongamia pinnata as standard. The 2C DNA content of RS coriander variety was estimated to 5.1 pg; the primary and secondary somatic embryo-derived plants had 5.26 and 5.44 pg 2C DNA content, respectively. The regenerated plants were genetically stable, genome size similar to seed-germinated coriander plants.

  13. Plant regeneration of non-toxic Jatropha curcas—impacts of plant growth regulators, source and type of explants

    KAUST Repository

    Kumar, Nitish

    2011-01-28

    Jatropha curcas is an oil bearing species with multiple uses and considerable economic potential as a biofuel plant, however, oil and deoiled cake are toxic. A non-toxic variety of J. curcas is reported from Mexico. The present investigation explores the effects of different plant growth regulators (PGRs) viz. 6-benzyl aminopurine (BAP) or thidiazuron (TDZ) individually and in combination with indole-3-butyric acid (IBA), on regeneration from in vitro and field-grown mature leaf explants, in vitro and glasshouse-grown seedlings cotyledonary leaf explants of non-toxic J. curcas. In all the tested parameters maximum regeneration efficiency (81.07%) and the number of shoot buds per explants (20.17) was observed on 9.08 μM TDZ containing Murashige and Skoog’s (MS) medium from in vitro cotyledonary leaf explants. The regenerated shoot buds were transferred to MS medium containing 10 μM kinetin (Kn), 4.5 μM BAP and 5.5 μM α-naphthaleneacetic acid (NAA) for shoot proliferation. The proliferated shoots could be elongated on MS medium supplemented with 2.25 μM BAP and 8.5 μM IAA. Rooting was achieved when the basal cut end of elongated shoots were dipped in half strength MS liquid medium containing different concentrations and combinations of IBA, IAA and NAA for four days followed by transfer to growth regulators free half strength MS medium supplemented 0.25 mg/l activated charcoal. The rooted plants could be established in soil with more than 90% survival rate.

  14. Influence of nutrient composition and plant growth regulators on callus induction and plant regeneration in glutinous rice (Oryza sativa L.).

    Science.gov (United States)

    Duangsee, K; Bunnag, S

    2014-01-01

    The potential for callus induction and regeneration depends on nutrient composition and plant growth regulators. The aim of the present study was to investigate the effect of nutrient composition and plant growth regulators on callus induction and plant regeneration in the glutinous rice cultivar Khunvang. The effect of 2,4-D concentrations (1, 2, 3, 4 and 5 mg L(-1)) on callus induction and growth were investigated. The results revealed that the highest percentage of callus induction (97%) was observed in MS medium supplemented with 5 mg L(-1) 2,4-D under 16 h Photoperiod. The effects of casein hydrolysate concentrations of casein hydrolysate (0, 300, 500, 700 and 900 mg L(-1)) and proline (0, 300, 500, 700 and 900 mg L(-1)) on callus induction and growth of Khunvang were also observed. The results indicated that the increasing casein hydrolysate and proline concentrations did not show a significant effect on callus growth. However, proline concentration of 900 mg L(-1) yielded 85.67% of callus growth.

  15. Inside out: high-efficiency plant regeneration and Agrobacterium-mediated transformation of upland and lowland switchgrass cultivars.

    Science.gov (United States)

    Liu, Yan-Rong; Cen, Hui-Fang; Yan, Jian-Ping; Zhang, Yun-Wei; Zhang, Wan-Jun

    2015-07-01

    Selection of pre-embryogenic callus from a core structure from mature seed-derived callus is the key for high-efficiency plant regeneration and transformation of switchgrass different cultivars. Switchgrass (Panicum virgatum L.) has been identified as a dedicated biofuel crop. For its trait improvement through biotechnological approaches, we have developed a highly efficient plant regeneration and genetic transformation protocol for both lowland and upland cultivars. We identified and separated a pre-embryogenic "core" structure from the seed-derived callus, which often leads to development of highly regenerative type II calluses. From the type II callus, plant regeneration rate of lowland cultivars Alamo and Performer reaches 95%, and upland cultivars Blackwell and Dacotah, 50 and 76%, respectively. The type II callus was also amenable for Agrobacterium-mediated transformation. Transformation efficiency of 72.8% was achieved for lowland cultivar Alamo, and 8.0% for upland cultivar Dacotah. PCR, Southern blot and GUS staining assays were performed to verify the transgenic events. High regenerative callus lines could be established in 3 months, and transgenic plants could be obtained in 2 months after Agrobacterium infection. To our knowledge, this is the first report on successful plant regeneration and recovery of transgenic plants from upland switchgrass cultivars by Agrobacterium-mediated transformation. The method presented here could be helpful in breaking through the bottleneck of regeneration and transformation of lowland and upland switchgrass cultivars and probably other recalcitrant grass crops.

  16. Methods for Rapid Screening in Woody Plant Herbicide Development

    Directory of Open Access Journals (Sweden)

    William Stanley

    2014-07-01

    Full Text Available Methods for woody plant herbicide screening were assayed with the goal of reducing resources and time required to conduct preliminary screenings for new products. Rapid screening methods tested included greenhouse seedling screening, germinal screening, and seed screening. Triclopyr and eight experimental herbicides from Dow AgroSciences (DAS 313, 402, 534, 548, 602, 729, 779, and 896 were tested on black locust, loblolly pine, red maple, sweetgum, and water oak. Screening results detected differences in herbicide and species in all experiments in much less time (days to weeks than traditional field screenings and consumed significantly less resources (<500 mg acid equivalent per herbicide per screening. Using regression analysis, various rapid screening methods were linked into a system capable of rapidly and inexpensively assessing herbicide efficacy and spectrum of activity. Implementation of such a system could streamline early-stage herbicide development leading to field trials, potentially freeing resources for use in development of beneficial new herbicide products.

  17. In vitro plant regeneration system for tropical butternut squash genotypes (Cucurbita moschata

    Directory of Open Access Journals (Sweden)

    Marta Valdez-Melara

    2009-11-01

    Full Text Available An efficient and reproducible method for regeneration of commercial and pure lines of tropical butternut squash (Cucurbita moschata plants via somatic embryogenesis was developed. The influence of genotype, explant source, N6-benzylaminopurine (BAP, 2,4-dichlorophenoxyacetic acid (2,4-D and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T concentration on somatic embryogenesis induction was investigated. Friable embryogenic calli was produced from zigotic embryos (53-56% and cotyledons from seedlings (70% of C. moschata cv. Sello de Oro cultured on callus induction medium (CIM supplemented with 0.5 mg/l or 3.5 mg/l 2,4-D. No embryogenic calli was obtained from leaf segments of C. moschata cv. Sello de Oro cultured on CIM supplemented with different concentrations of BAP and 2,4-D and cotyledons from seedlings of C. moschata cv. PVG 04 cultured on CIM with BAP and 2,4,5-T. Embryogenic calli induction was achieved in 75% C. moschata pure lines evaluated and calli percentage frequency range from 5% to 34%. Successful acclimatization of squash in vitro plants was achieved in the greenhouse and in the field. Regenerated plants appeared morphologically normal and set flowers and fruits with seeds that could germinate normally. Rev. Biol. Trop. 57 (Suppl. 1: 119-127. Epub 2009 November 30.

  18. The caudal regeneration blastema is an accumulation of rapidly proliferating stem cells in the flatworm Macrostomum lignano

    Directory of Open Access Journals (Sweden)

    Adamski Zbigniew

    2009-07-01

    Full Text Available Abstract Background Macrostomum lignano is a small free-living flatworm capable of regenerating all body parts posterior of the pharynx and anterior to the brain. We quantified the cellular composition of the caudal-most body region, the tail plate, and investigated regeneration of the tail plate in vivo and in semithin sections labeled with bromodeoxyuridine, a marker for stem cells (neoblasts in S-phase. Results The tail plate accomodates the male genital apparatus and consists of about 3,100 cells, about half of which are epidermal cells. A distinct regeneration blastema, characterized by a local accumulation of rapidly proliferating neoblasts and consisting of about 420 cells (excluding epidermal cells, was formed 24 hours after amputation. Differentiated cells in the blastema were observed two days after amputation (with about 920 blastema cells, while the male genital apparatus required four to five days for full differentiation. At all time points, mitoses were found within the blastema. At the place of organ differentiation, neoblasts did not replicate or divide. After three days, the blastema was made of about 1420 cells and gradually transformed into organ primordia, while the proliferation rate decreased. The cell number of the tail plate, including about 960 epidermal cells, was restored to 75% at this time point. Conclusion Regeneration after artificial amputation of the tail plate of adult specimens of Macrostomum lignano involves wound healing and the formation of a regeneration blastema. Neoblasts undergo extensive proliferation within the blastema. Proliferation patterns of S-phase neoblasts indicate that neoblasts are either determined to follow a specific cell fate not before, but after going through S-phase, or that they can be redetermined after S-phase. In pulse-chase experiments, dispersed distribution of label suggests that S-phase labeled progenitor cells of the male genital apparatus undergo further proliferation before

  19. Plant regeneration through callus organogenesis and true-to-type conformity of plants by RAPD analysis in Desmodium gangeticum (Linn.) DC.

    Science.gov (United States)

    Cheruvathur, Meena K; Abraham, Jyothi; Thomas, T Dennis

    2013-03-01

    An efficient plant regeneration protocol was established for an endangered ethnomedicinal plant Desmodium gangeticum (Linn.) DC. Morphogenic calli were produced from 96 % of the cultures comprising the immature leaf explants on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (4.0 mg l(-1)) in combination with 6-benzylaminopurine (BA; 0.8 mg l(-1)). For callus regeneration, various concentrations of BA (1.0-5.0 mg l(-1)) or thidiazuron (TDZ; 1.0-5.0 mg l(-1)) alone or in combination with indole-3-acetic acid (IAA; 0.2-1.0 mg l(-1)) were used. Highest response of shoot regeneration was observed on MS medium fortified with TDZ (4.0 mg l(-1)) and IAA (0.5 mg l(-1)) combination. Here, 100 % cultures responded with an average number of 22.3 shoots per gram calli. Inclusion of indole-3-butyric acid in half MS medium favored rooting of recovered shoots. Out of 45 rooted plants transferred to soil, 40 survived. Total DNA was extracted from the leaves of the acclimatized plants of D. gangeticum. Analysis of random amplified polymorphic DNA using 13 arbitrary decanucleotide primers showed the genetic homogeneity in all the ten plants regenerated from callus with parental plant, suggesting that shoot regeneration from callus could be used for the true-to-type multiplication of this plant.

  20. High Frequency Plant Regeneration System from Transverse Thin Cell Layer Section of In vitro Derived ‘Nadia’ Ginger Microrhizome

    Directory of Open Access Journals (Sweden)

    Dikash Singh THINGBAIJAM

    2014-03-01

    Full Text Available An efficient and reproducible procedure is outlined for rapid in vitro multiplication of Zingiber officinale var. ‘Nadia’ through high frequency shoot proliferation from transverse thin cell layer (tTCL sections of in vitro derived microrhizome. In vitro derived microrhizome of size 500 μm in thickness was used as initial explants for induction of somatic embryos. Among the different phytohormones tested, tTCL explants shows maximum calli proliferation in medium containing 2 mg/L 2,4-Dichlorophenoxyacetic acid (88.30±0.11%. Reduced concentration of 2,4 Dichlorophenoxyacetic acid was supplemented with different cytokinins for regeneration of callus. Among the different medium tested, optimum redifferentiation of somatic embryos were observed in medium containing 0.2 mg/L 2,4 Dichlorophenoxyacetic acid and 6.0 mg/L BAP (141.08±0.25. Clump of regenerated plantlets were further subculture and transfer into microrhizome inducing medium containing high sucrose concentration (8%. Plantlets with well developed microrhizome were successfully acclimatized and eventually transferred to the field. The application of studying embryo section for regeneration of plants might be useful alternative to ginger improvement programme. Histological analysis showed formation of somatic embryos and regenerated adventitious shoot.

  1. Genetic stability and phytochemical analysis of the in vitro regenerated plants of Dendrobium nobile Lindl., an endangered medicinal orchid

    Science.gov (United States)

    Bhattacharyya, Paromik; Kumaria, Suman; Diengdoh, Reemavareen; Tandon, Pramod

    2014-01-01

    An efficient genetically stable regeneration protocol with increased phytochemical production has been established for Dendrobium nobile, a highly prized orchid for its economic and medicinal importance. Protocorm like bodies (PLBs) were induced from the pseudostem segments using thidiazuron (TDZ; 1.5 mg/l), by-passing the conventional auxin–cytokinin complement approach for plant regeneration. Although, PLB induction was observed at higher concentrations of TDZ, plantlet regeneration from those PLBs was affected adversely. The best rooting (5.41 roots/shoot) was achieved in MS medium with 1.5 mg/l TDZ and 0.25% activated charcoal. Plantlets were successfully transferred to a greenhouse with a survival rate of 84.3%, exhibiting normal development. Genetic stability of the regenerated plants was investigated using randomly amplified polymorphic DNA (RAPD) and start codon targeted (SCoT) polymorphism markers which detected 97% of genetic fidelity among the regenerants. The PIC values of RAPD and SCoT primers were recorded to be 0.92 and 0.76 and their Rp values ranged between 3.66 and 10, and 4 and 12 respectively. The amplification products of the regenerated plants showed similar banding patterns to that of the mother plant thus demonstrating the homogeneity of the micropropagated plants. A comparative phytochemical analysis among the mother and the micropropagated plants showed a higher yield of secondary metabolites. The regeneration protocol developed in this study provides a basis for ex-situ germplasm conservation and also harnesses the various secondary metabolite compounds of medicinal importance present in D. nobile. PMID:25606433

  2. Early development of matched planted and naturally regenerated Douglas-fir stands after slash burning in the Cascade Range.

    Science.gov (United States)

    R.E. Miller; R.E. Bigley; S. Webster

    1993-01-01

    We compared matched planted and naturally regenerated plots in 35- to 38- year-old Douglas-fir (Pseudotsuga menziesii var. menziesii) stands at seven locations in western Washington and Oregon. Total number of live stems is similar, but stands planted to Douglas fir average 26 more live stemslac of Douglas-fir and 39 fewer...

  3. Clearcutting and Site Preparation, but Not Planting, Promoted Early Tree Regeneration in Boreal Alaska

    Directory of Open Access Journals (Sweden)

    Miho Morimoto

    2016-12-01

    Full Text Available The stand initiation stage decisively influences future forest structure and composition, particularly in the boreal forest which is a stand replacement disturbance driven system. In boreal Alaska, the conventional forest management paradigm has focused on the production of large-dimension timber, particularly white spruce (Picea glauca. However, energy generation and heating from wood is increasing, and is likely to significantly expand total forest harvest, further shifting management focus to fuelwood production. We evaluated the effects of forest harvest management practices on post-harvest regeneration by examining whether harvest type, site preparation method, and reforestation technique resulted in differences in forest regeneration in terms of species presence, dominance, basal area, and total stem biomass using a stochastic gradient boosting (TreeNet algorithm. We recorded diameter at breast height and height of white spruce, birch (Betula neoalaskana, and aspen (Populus tremuloides in 726 plots from 30 harvest units, distributed across the various harvest and treatment types, harvest years, harvest sizes, and geographical locations. Our results indicate that management practices suitable/acceptable for woody biomass production differ from the more traditional dimensional timber production from white spruce-focused management. Artificial reforestation does not differ from natural regeneration in obtaining more stems or producing greater biomass. Clearcutting and site preparation increased tree regeneration, basal area, and woody biomass when compared to a partial harvest with no site preparation. Planting of white spruce in the Alaskan boreal forest may only be necessary in some specific circumstances, such as years with no/low white spruce seed crop, or in landscapes depleted of seed trees.

  4. Seismomorphogenesis: a novel approach to acclimatization of tissue culture regenerated plants.

    Science.gov (United States)

    Sarmast, Mostafa Khoshhal; Salehi, Hassan; Khosh-Khui, Morteza

    2014-12-01

    Plantlets under in vitro conditions transferred to ex vivo conditions are exposed to biotic and abiotic stresses. Furthermore, in vitro regenerated plants are typically frail and sometimes difficult to handle subsequently increasing their risk to damage and disease; hence acclimatization of these plantlets is the most important step in tissue culture techniques. An experiment was conducted under in vitro conditions to study the effects of shaking duration (twice daily at 6:00 a.m. and 9:00 p.m. for 2, 4, 8, and 16 min at 250 rpm for 14 days) on Sansevieria trifasciata L. as a model plant. Results showed that shaking improved handling, total plant height, and leaf characteristics of the model plant. Forty-eight hours after 14 days of shaking treatments with increasing shaking time, leaf length decreased but proline content of leaf increased. However, 6 months after starting the experiment different results were observed. In explants that received 16 min of shaking treatment, leaf length and area and photosynthesis rate were increased compared with control plantlets. Six months after starting the experiment, control plantlets had 12.5 % mortality; however, no mortality was observed in other treated explants. The results demonstrated that shaking improved the explants' root length and number and as a simple, cost-effective, and non-chemical novel approach may be substituted for other prevalent acclimatization techniques used for tissue culture regenerated plantlets. Further studies with sensitive plants are needed to establish this hypothesis.

  5. Uptake, effects, and regeneration of barley plants exposed to gold nanoparticles.

    Science.gov (United States)

    Feichtmeier, Nadine S; Walther, Paul; Leopold, Kerstin

    2015-06-01

    Within the last years, nanogold has become more and more important in nanotechnology, for example, as catalyst or in medical applications. Its rising production, application, and disposal inevitably lead to an increased emission of gold nanoparticles (Au-NPs) in the environment. However, only little is known about the uptake and effects of Au-NPs on biota. The objective of this study was to investigate the reversibility and effects of citrate-coated Au-NP uptake into the model organism barley (Hordeum vulgare L.). For this purpose, barley seeds were cultivated in Au-NP-containing nutrient solution for 2 weeks before the seedlings were transferred into Au-NP-free media and grown for another 3 weeks. Stability of Au-NPs in the cultivation media was investigated over the 2-week exposure time. Gold content in the leaves and roots of the plants was measured after 2 weeks of exposure and after 7, 14, and 21 days of regeneration by means of total reflection X-ray fluorescence (TXRF) analysis after microwave-assisted digestion. Moreover, Au-NPs within plant material were localized by transmission electron microscopy (TEM) of ultrathin cross sections. The obtained results reveal that Au-NPs accumulate in the plant roots. Concentration-dependent effects on the uptake of macronutrients and micronutrients, as well as on biomass production of exposed plants, in particular, on root growth were observed. Even though exposed barley plants were able to regenerate to a certain extent, their root growth was permanently decreased.

  6. Plant regeneration via somatic embryogenesis and shoot organogenesis from immature cotyledons of Camellia nitidissima Chi.

    Science.gov (United States)

    Lü, Jinfeng; Chen, Rong; Zhang, Muhan; da Silva, Jaime A Teixeira; Ma, Guohua

    2013-09-01

    Camellia nitidissima Chi (Theaceae) is a world-famous economic and ornamental plant with golden-yellow flowers. It has been classified as one of the rarest and most endangered plants in China. Our objective was to induce somatic embryogenesis, shoot organogenesis and plant regeneration for C. nitidissima. Three types of callus (whitish, reddish and yellowish) were induced from immature cotyledons on improved woody plant medium (WPM) with different plant growth regulators (PGRs). Among the callus, whitish callus was induced by 4.5 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and reddish and yellowish callus were induced by strongly active cytokinins, thidiazuron (TDZ) or 6-benzylaminopurine (BAP), singly or combined with weakly active auxin, α-naphthaleneacetic acid (NAA). The embryogenic callus could differentiate into somatic embryos, nodular embryogenic structures (large embryo-like structures) or adventitious shoots depending on the PGR used in WPM. BAP was best for adventitious buds and zeatin was best for somatic embryogenesis while kinetin (Kt) was best for the formation of nodular embryogenic structures. The three regeneration pathways often occurred in the same embryogenic callus clumps. Most shoots (80.0%) developed roots in WPM supplemented with 24.6 μM IBA and 0.3 μM NAA while 47.5% of somatic embryos could germinate directly and develop into plantlets on induction medium supplemented with 0.9 μM BAP and 0.1 μM NAA. The nodular embryogenic structures could be sub-cultured and cyclically developed in one of two differentiation pathways: shoot organogenesis or somatic embryogenesis. Plantlets derived from shoot buds rooted and somatic embryos germinated when transplanted into soil in a greenhouse; 66.7% of plantlets from shoot culture and 78.6% of plantlets from somatic embryos survived after 8 weeks' acclimatization. Copyright © 2013 Elsevier GmbH. All rights reserved.

  7. Integrated bicarbonate-form ion exchange treatment and regeneration for DOC removal: Model development and pilot plant study.

    Science.gov (United States)

    Hu, Yue; Boyer, Treavor H

    2017-05-15

    The application of bicarbonate-form anion exchange resin and sodium bicarbonate salt for resin regeneration was investigated in this research is to reduce chloride ion release during treatment and the disposal burden of sodium chloride regeneration solution when using traditional chloride-form ion exchange (IX). The target contaminant in this research was dissolved organic carbon (DOC). The performance evaluation was conducted in a completely mixed flow reactor (CMFR) IX configuration. A process model that integrated treatment and regeneration was investigated based on the characteristics of configuration. The kinetic and equilibrium experiments were performed to obtain required parameters for the process model. The pilot plant tests were conducted to validate the model as well as provide practical understanding on operation. The DOC concentration predicted by the process model responded to the change of salt concentration in the solution, and showed a good agreement with pilot plant data with less than 10% difference in terms of percentage removal. Both model predictions and pilot plant tests showed over 60% DOC removal by bicarbonate-form resin for treatment and sodium bicarbonate for regeneration, which was comparable to chloride-form resin for treatment and sodium chloride for regeneration. Lastly, the DOC removal was improved by using higher salt concentration for regeneration. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Biotechnological applications in in vitro plant regeneration studies of broccoli (Brassica oleracea L. var. italica), an important vegetable crop.

    Science.gov (United States)

    Kumar, Pankaj; Srivastava, Dinesh Kumar

    2016-04-01

    Biotechnology holds promise for genetic improvement of important vegetable crops. Broccoli (Brassica oleracea L. var. italica) is an important vegetable crop of the family Brassicaceae. However, various biotic and abiotic stresses cause enormous crop yield losses during commercial cultivation of broccoli. Establishment of a reliable, reproducible and efficient in vitro plant regeneration system with cell and tissue culture is a vital prerequisite for biotechnological application of crop improvement programme. An in vitro plant regeneration technique refers to culturing, cell division, cell multiplication, de-differentiation and differentiation of cells, protoplasts, tissues and organs on defined liquid/solid medium under aseptic and controlled environment. Recent progress in the field of plant tissue culture has made this area one of the most dynamic and promising in experimental biology. There are many published reports on in vitro plant regeneration studies in broccoli including direct organogenesis, indirect organogenesis and somatic embryogenesis. This review summarizes those plant regeneration studies in broccoli that could be helpful in drawing the attention of the researchers and scientists to work on it to produce healthy, biotic and abiotic stress resistant plant material and to carry out genetic transformation studies for the production of transgenic plants.

  9. Plant diversity and regeneration in a disturbed isolated dry Afromontane forest in northern Ethiopia

    DEFF Research Database (Denmark)

    Aynekulu, Ermias; Aerts, Raf; Denich, Manfred

    2016-01-01

    collectively interpreted as a moisture gradient, and forest disturbance separated the plant communities. With only 39 of the 79 recorded woody species present in the seedling layer, the forest currently faces an extinction debt of 50 per cent of the total woody species pool. Human disturbance has clearly......We studied the diversity, community composition and natural regeneration of woody species in an isolated but relatively large (> 1,000 ha) dry Afromontane forest in northern Ethiopia to assess its importance for regional forest biodiversity conservation. The principal human-induced disturbance...... regimes affecting this forest include logging and livestock grazing. Vegetation data were collected in 65 plots (50 m × 50 m); seedling species composition and density were determined in 10 m × 10 m nested plots. We used a cluster analysis to identify plant communities and non-metric multidimensional...

  10. High Frequency of Plant Regeneration through Cyclic Secondary Somatic Embryogenesis in Panax ginseng.

    Science.gov (United States)

    Kim, Yu-Jin; Lee, Ok Ran; Kim, Kyung-Tack; Yang, Deok-Chun

    2012-10-01

    Somatic embryogenesis is one of good examples of the basic research for plant embryo development as well as an important technique for plant biotechnology such as medicinally important plants. Single embryos develop into normal plantlets with shoots and roots. Therefore, direct single embryogenesis derived from single cells is highly important for normal plant regeneration. Here we demonstrate that the cyclic secondary somatic embryogenesis in Panax ginseng Meyer is a permanent source of embryogenic material that can be used for genetic manipulations. Secondary somatic embryos were originated directly from the primary somatic embryos on hormone-free Murashige and Skoog medium, and proliferated further in a cyclic manner. EM medium (one third of modified MS medium [MS medium containing half amount of NH4NO3 and KNO3] with 2% to 3% sucrose) favored further development of proliferated secondary somatic embryos into plantlets with root system. The plantlets developed into plants with well-developed taproots in half-strength Schenk and Hildebrandt basal medium supplemented with 0.5% activated charcoal.

  11. Rapid Control Prototyping Plataform for Didactic Plant Motor DC

    Directory of Open Access Journals (Sweden)

    Cristian Bazán-Orobio

    2013-06-01

    Full Text Available In this paper a design, implementation and validation of a Rapid Control Prototype platform for a plant based on a DC motor is proposed. This low-cost prototype provides of an electronic card (with a motor DC and sensors manipulated by PC with free software tools using Linux, Scilab / Scicos and RTAI-Lab. This RCP System allows developing speed -position control trainings by using different types of PID industrial controllers with anti – wind up and bump less transfer schemes. We develop a speed control application structured in four steps: identification, controller design, simulation and real time control, where there are pedagogical advantages of a platform that not only allows simulation but also real-time control of a plant.

  12. Phenotypic and molecular characterization of plants regenerated from non-cryopreserved and cryopreserved wild Solanum lycopersicum mill. Seeds.

    Science.gov (United States)

    Zevallos, B; Cejas, I; Engelmann, F; Carputo, D; Aversano, R; Scarano, M T; Yanes, E; Martinez-Montero, M; Lorenzo, J C

    2014-01-01

    Before cryopreservation is routinely used, its effect on the trueness-to-type of the regenerated plant material needs to be evaluated. In this work, we studied the effect of seed cryopreservation on the phenotypic and molecular characteristics of wild Solanum lycopersicum Mill. plants. Thirty-five morphological traits of plants regenerated from cryopreserved seeds were compared to those measured on plants regenerated from non-cryopreserved seeds. No statistically significant differences were observed between cryopreserved and non-cryopreserved samples, either in the first or in the second generation post-liquid nitrogen exposure. However, at the molecular level, the genetic analyses performed on the second generation plants germinated from control and cryopreserved seeds using 14 nuclear Simple Sequences Repeats (SSR) markers uncovered some changes in microsatellite length between control and cryopreserved samples. These results confirm at the botanical phenotype level the effectiveness of seed cryostorage for conservation and regeneration of true-to-type S. lycopersicum plants. Further experiments are required to clarify potential phenotypic effects of the changes observed in the DNA.

  13. Rapid evolution of manifold CRISPR systems for plant genome editing

    Directory of Open Access Journals (Sweden)

    Yiping Qi

    2016-11-01

    Full Text Available Advanced CRISPR-Cas9 based technologies first validated in mammalian cell systems are quickly being adapted for use in plants. These new technologies increase CRISPR-Cas9’s utility and effectiveness by diversifying cellular capabilities through expression construct system evolution and enzyme orthogonality, as well as enhanced efficiency through delivery and expression mechanisms. Here, we review the current state of advanced CRISPR-Cas9 and Cpf1 capabilities in plants and cover the rapid evolution of these tools from first generation inducers of double strand breaks for basic genetic manipulations to second and third generation multiplexed systems with myriad functionalities, capabilities and specialized applications. We offer perspective on how to utilize these tools for currently untested research endeavors and analyze strengths and weaknesses of novel CRISPR systems in plants. Advanced CRISPR functionalities and delivery options demonstrated in plants are primarily reviewed but new technologies just coming to the forefront of CRISPR development, or those on the horizon, are briefly discussed. Topics covered are focused on the expansion of expression and delivery capabilities for CRISPR-Cas9 components and broadening targeting range through orthogonal Cas9 and Cpf1 proteins.

  14. Cross-resistance of cell lines and plant regenerants of winter triticale to abiotic stressors

    Directory of Open Access Journals (Sweden)

    С. В. Пикало

    2017-12-01

    Full Text Available Purpose. To analyze the level of cross-resistance of obtained salt- and osmotolerant cell lines and plants regenerants of winter triticale to osmotic and salt stresses. Methods. Cultures of tissue and organs in vitro, in vitro breeding, biochemical, statistical analysis. Results. It was established that the stability of cross-resistance trait display to saline and osmotic stresses in obtained cell lines of winter triticale was rather high – from 50 to 76% of calli have survived to the end of the sixth passage. It has been shown that despite the presence of sublethal concentrations of the stress-factor (mannitol/sodium chloride in selective medium, stable cell lines of the triticale actively continued to grow and accumulate biomass. It was found that in the line ‘38/1296’ cell lines 5L/sl and 5L/os respectively were the most resistant to osmotic and salt stresses, and lines 1C/s1 and 1C/os respectively in the ‘Obrii’ variety, since they had the highest percent of living calli and biomass increment under the selective conditions and their plant regenerant – the highest level of survival after the impact of the abiotic stressors complex. The salt-resistant cell lines of both genotypes of winter triticale as compared to the control were also characterized by significantly higher free proline content under the selective factors impact. The results obtained may indicate that the cell lines and triticale plant regenerants have a genetically determined trait of resistance to stress factors. Conclusions. Verification of traits of resistance to abiotic stressors has shown a significantly high level of cross-tolerance of the obtained cell lines of both triticale genotypes for saline and osmotic stresses. Resistance to saline and osmotic stresses of cells separated in vitro was preserved in induced plants and at the organism level has increased tolerance to abiotic environmental factors. It is shown that due to the general non

  15. Studies on Callus Induction and Regeneration of Medicinal Plant Chicory (Cichorium intybus L. from Leaf and Petiole Explants

    Directory of Open Access Journals (Sweden)

    H. Hadizadeh

    2016-07-01

    Full Text Available Introduction: Chicory (Cichorium intybus L. belongs to Asteraceae family is commonly known as witloof chicory. The leaves and the roots of this medicinal plant are edible and commonly used as salad. Some varieties are also cultivated as coffee substitute after roasting the roots. All parts of the plant contain these volatile oils, with the majority of the toxic components concentrated in the plant's root. In folk medicine, the plant is used for the treatment of diarrhea, spleen enlargement, fever, and vomiting. Antihepatotoxic activity on damaged rat’s liver sections and anti-bacterial activity of this crop has been recently reported. In vitro regeneration from leaf explants with various hormonal combinations has been reported previously. Moreover, in vitro regeneration of Chicory from cotyledon explants using different combinations of plant growth regulators has been studied. Also, a protocol for the regeneration of plantlets from leaf and petiole explants of witloof chicory has been developed. The aim of the present investigation was optimization of callus induction and shoot regeneration from leaf and petiole tissues of Chicory (Esfahan genotype. Materials and Methods: In this investigation, Esfahan genotype was used for callus induction and direct shoot regeneration. Seeds were first washed with running tap water for 30 min then seeds were surface sterilized by dipping in 70% ethanol for 90 s and rinsed with sterile distilled water, followed by immersing in 5% sodium hypochlorite solution for 25 min and thereafter rinsed for 30 min with sterile distilled water. The basal medium used in this investigation was MS. For shoot regeneration, leaf and petiole explants (5 mm segments were excised from 4-week-old sterile seedlings and cultured on MS medium containing different combinations of NAA / BA and KIN / BA in two separate experiments. Experiments were performed factorial based on completely randomized design. Cultures were incubated at 25

  16. Effects of high-frequency understorey fires on woody plant regeneration in southeastern Amazonian forests

    Science.gov (United States)

    Balch, Jennifer K.; Massad, Tara J.; Brando, Paulo M.; Nepstad, Daniel C.; Curran, Lisa M.

    2013-01-01

    Anthropogenic understorey fires affect large areas of tropical forest, yet their effects on woody plant regeneration post-fire remain poorly understood. We examined the effects of repeated experimental fires on woody stem (less than 1 cm at base) mortality, recruitment, species diversity, community similarity and regeneration mode (seed versus sprout) in Mato Grosso, Brazil. From 2004 to 2010, forest plots (50 ha) were burned twice (B2) or five times (B5), and compared with an unburned control (B0). Stem density recovered within a year after the first burn (initial density: 12.4–13.2 stems m−2), but after 6 years, increased mortality and decreased regeneration—primarily of seedlings—led to a 63 per cent and 85 per cent reduction in stem density in B2 and B5, respectively. Seedlings and sprouts across plots in 2010 displayed remarkable community similarity owing to shared abundant species. Although the dominant surviving species were similar across plots, a major increase in sprouting occurred—almost three- and fourfold greater in B2 and B5 than in B0. In B5, 29 species disappeared and were replaced by 11 new species often present along fragmented forest edges. By 2010, the annual burn regime created substantial divergence between the seedling community and the initial adult tree community (greater than or equal to 20 cm dbh). Increased droughts and continued anthropogenic ignitions associated with frontier land uses may promote high-frequency fire regimes that may substantially alter regeneration and therefore successional processes. PMID:23610167

  17. Thidiazuron: A potent cytokinin for efficient plant regeneration in Himalayan poplar (Populus ciliata Wall. using leaf explants

    Directory of Open Access Journals (Sweden)

    Gaurav Aggarwal

    2012-11-01

    Full Text Available Populus species are important resource for certain branches of industry and have special roles for scientific study on biological and agricultural systems. The present investigation was undertaken with an objective of enhancing the frequency of plant regeneration in Himalayan poplar (Populus ciliata Wall.. The effect of Thiadizuron (TDZ alone and in combination with adenine and α-Naphthalene acetic acid (NAA were studied on the regeneration potential of leaf explants. A high efficiency of shoot regeneration was observed in leaf (80.00% explants on MS basal medium supplemented with 0.024 mg/l TDZ and 79.7 mg/l adenine. Elongation and multiplication of shoots were obtained on Murashige and Skoog (MS basal medium, containing 0.5 mg/l 6. Benzyl aminopurine (BAP + 0.2mg/l Indole 3-acetic acid (IAA + 0.3 mg/l Gibberellic acid (GA3. High frequency root regeneration from in vitro developed shoots was observed on MS basal medium supplemented with 0.10 mg/l Indole 3-butyric acid(IBA. Maximum of the in vitro rooted plantlets were well accomplished to the mixture of sand: soil (1:1 and exhibited similar morphology with the field plants. A high efficiency plant regeneration protocol has been developedfrom leaf explants in Himalayan poplar (Populus ciliata Wall..

  18. Thidiazuron: A potent cytokinin for efficient plant regeneration in Himalayan poplar (Populus ciliata Wall. using leaf explants

    Directory of Open Access Journals (Sweden)

    Gaurav Aggarwal

    2012-12-01

    Full Text Available Populus species are important resource for certain branches of industry and have special roles for scientific study on biological and agricultural systems. The present investigation was undertaken with an objective of enhancing the frequency of plant regeneration in Himalayan poplar (Populus ciliataWall.. The effect of Thiadizuron (TDZ alone and in combination with adenine and alpha-Naphthalene acetic acid (NAA were studied on the regeneration potential of leaf explants. A high efficiency of shoot regeneration was observed in leaf (80.00% explants on MS basal medium supplemented with 0.024 mg/l TDZ and 79.7 mg/l adenine. Elongation and multiplication of shoots were obtained on Murashige and Skoog (MS basal medium, containing 0.5 mg/l 6. Benzyl aminopurine (BAP + 0.2mg/l Indole 3-acetic acid (IAA + 0.3 mg/l Gibberellic acid (GA3. High frequency root regeneration from in vitro developed shoots was observed on MS basal medium supplemented with 0.10 mg/l Indole 3-butyric acid (IBA. Maximum of the in vitro rooted plantlets were well accomplished to the mixture of sand: soil (1:1 and exhibited similar morphology with the field plants. A high efficiency plant regeneration protocol has been developed from leaf explants in Himalayan poplar (Populus ciliata Wall.. 

  19. Regeneration of viable oil palm plants from protoplasts by optimizing media components, growth regulators and cultivation procedures.

    Science.gov (United States)

    Masani, Mat Yunus Abdul; Noll, Gundula; Parveez, Ghulam Kadir Ahmad; Sambanthamurthi, Ravigadevi; Prüfer, Dirk

    2013-09-01

    Oil palm protoplasts are suitable as a starting material for the production of oil palm plants with new traits using approaches such as somatic hybridization, but attempts to regenerate viable plants from protoplasts have failed thus far. Here we demonstrate, for the first time, the regeneration of viable plants from protoplasts isolated from cell suspension cultures. We achieved a protoplast yield of 1.14×10(6) per gram fresh weight with a viability of 82% by incubating the callus in a digestion solution comprising 2% cellulase, 1% pectinase, 0.5% cellulase onuzuka R10, 0.1% pectolyase Y23, 3% KCl, 0.5% CaCl2 and 3.6% mannitol. The regeneration of protoplasts into viable plants required media optimization, the inclusion of plant growth regulators and the correct culture technique. Microcalli derived from protoplasts were obtained by establishing agarose bead cultures using Y3A medium supplemented with 10μM naphthalene acetic acid, 2μM 2,4-dichlorophenoxyacetic acid, 2μM indole-3-butyric acid, 2μM gibberellic acid and 2μM 2-γ-dimethylallylaminopurine. Small plantlets were regenerated from microcalli by somatic embryogenesis after successive subculturing steps in medium with limiting amounts of growth regulators supplemented with 200mg/l ascorbic acid. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  20. Rapid breeding of parthenocarpic tomato plants using CRISPR/Cas9.

    Science.gov (United States)

    Ueta, Risa; Abe, Chihiro; Watanabe, Takahito; Sugano, Shigeo S; Ishihara, Ryosuke; Ezura, Hiroshi; Osakabe, Yuriko; Osakabe, Keishi

    2017-03-30

    Parthenocarpy in horticultural crop plants is an important trait with agricultural value for various industrial purposes as well as direct eating quality. Here, we demonstrate a breeding strategy to generate parthenocarpic tomato plants using the CRISPR/Cas9 system. We optimized the CRISPR/Cas9 system to introduce somatic mutations effectively into SlIAA9-a key gene controlling parthenocarpy-with mutation rates of up to 100% in the T0 generation. Furthermore, analysis of off-target mutations using deep sequencing indicated that our customized gRNAs induced no additional mutations in the host genome. Regenerated mutants exhibited morphological changes in leaf shape and seedless fruit-a characteristic of parthenocarpic tomato. And the segregated next generation (T1) also showed a severe phenotype associated with the homozygous mutated genome. The system developed here could be applied to produce parthenocarpic tomato in a wide variety of cultivars, as well as other major horticultural crops, using this precise and rapid breeding technique.

  1. Thidiazuron-induced plant regeneration from protoplasts of Vicia faba cv. Mythos.

    Science.gov (United States)

    Tegeder, M; Gebhardt, D; Schieder, O; Pickardt, T

    1995-12-01

    Protoplasts of 10 cultivars of V. faba were isolated from etiolated shoot-tips and tested for their regeneration capacity. After purification, protoplasts were embedded in sodium alginate and cultivated in the medium of Kao and Michayluk (1975) containing 0.5 mg·1(-1) of each 2,4-dichlorophenoxyacetic acid, naphthylacetic acid and 6-benzylaminopurine. Depending on cultivar, division frequencies of up to 40% were obtained. Six weeks after embedding, protoplast-derived calluses were transferred to Gelrite-solidified media with different combinations of growth regulators. A two step protocol (auxin high/low) was tested for its ability to induce somatic embryogenesis. The formation of globular structures was observed, but no embryo formation could be achieved. In contrast, cultivation of protocalluses on medium supplemented with thidiazuron resulted in shoot development in cultivar Mythos. To generate mature plants, the shoots were grafted onto young seedlings. In order to optimize the in vitro-conditions, different concentrations of thidiazuron alone or in combination with naphthylacetic acid were tested, showing that an increase of thidiazuron and the addition of naphthylacetic acid positively affects both the viability of protocalluses and the regeneration frequency.

  2. Area Expansivity Moduli of Regenerating Plant Protoplast Cell Walls Exposed to Shear Flows

    Science.gov (United States)

    Fujimura, Yuu; Iino, Masaaki; Watanabe, Ugai

    2005-05-01

    To control the elasticity of the plant cell wall, protoplasts isolated from cultured Catharanthus roseus cells were regenerated in shear flows of 115 s-1 (high shear) and 19.2 s-1 (low shear, as a control). The surface area expansivity modulus and the surface breaking strength of these regenerating protoplasts were measured by a micropipette aspiration technique. Cell wall synthesis was also measured using a cell wall-specific fluorescent dye. High shear exposure for 3 h doubled both the surface area modulus and breaking strength observed under low shear, significantly decreased cell wall synthesis, and roughly quadrupled the moduli of the cell wall. Based on the cell wall synthesis data, we estimated the three-dimensional modulus of the cell wall to be 4.1± 1.2 GPa for the high shear, and 0.35± 0.2 GPa for the low shear condition, using the surface area expansivity modulus divided by the cell wall thickness, which is identical with the Young’s modulus divided by 2(1-σ), where σ is Poisson's ratio. We concluded that high shear exposure considerably strengthens the newly synthesized cell wall.

  3. Modified AFLP technique for rapid genetic characterization in plants.

    Science.gov (United States)

    Ranamukhaarachchi, D G; Kane, M E; Guy, C L; Li, Q B

    2000-10-01

    The standard amplified fragment-length polymorphism (AFLP) technique was modified to develop a convenient and reliable technique for rapid genetic characterization of plants. Modifications included (i) using one restriction enzyme, one adapter molecule and primer, (ii) incorporating formamide to generate more intense and uniform bands and (iii) using agarose gel electrophoresis. Sea oats (Uniola paniculata L.), pickerel-weed (Pontederia cordata L.), Bermudagrass (Cynodon dactylon L.) and Penstemon heterophyllus Lindl. were used to determine the ability to generate adequate resolution power with both self- and cross-pollinated plant species including cultivars, ecotypes and individuals within populations. Reproducibility of bands was higher in all the AFLP experiments compared to random amplified polymorphic DNA (RAPD). Formamide with or without bovine serum albumin improved band intensities compared to dimethyl sulfoxide and the standard reaction mixture with no organic solvents. Comparison between RAPD and modified AFLP using sea-oats population samples proved that modified AFLP exhibits (i) a low number of faint bands with increased specificity of amplified bands, (ii) a significantly higher number of polymorphic loci per primer, (iii) less primer screening time, (iv) easy scoring associated with fewer faint bands and (v) greatly enhanced reproducibility. The technique described here can be applied with a high degree of accuracy for plant genetic characterization.

  4. Callus induction and plant regeneration from explants of commercial cultivars of leek (Allium ampeloprasum var. porrum L.).

    Science.gov (United States)

    Buiteveld, J; van der Valk, P; Jansen, J; Creemers-Molenaar, J; Colijn-Hooymans, C M

    1993-05-01

    Plant regeneration capacity was studied for 8 cultivars and 4 accessions of leek (A. ampeloprasum var. porrum L.). Compact callus was induced on embryo and leaf explants on three different media. The highest frequency of compact callus formation (up to 90%) was obtained when mature, zygotic embryos were cultured on MS medium, containing 30 g/l sucrose and 1 mg/l 2,4-D. Regeneration occurred through somatic embryogenesis on MS medium, supplemented with 1 mg/l kinetin. Plants could be regenerated from all cultivars and accessions tested. These cultivars and accessions could be classified into three groups with respect to shoot formation frequency. The results suggest a distinct influence of the genotype on the morphogenic response of leek embryo explants in vitro.

  5. Rapid Screening of In-Vitro Regenerated Plantlets of Four Nigerian ...

    African Journals Online (AJOL)

    Prof. Ogunji

    pathogen interactions .... It is left to plant breeders to investigate the heritability of this positive trait. But as advances are made in mutation ... enhanced resistance to fungal pathogen and insect pest. In: Brady, N.C (ed.) Advances in Agronomy.

  6. Regeneration of whole plants of geranium from petioles cultured in vitro

    Directory of Open Access Journals (Sweden)

    Barbara Stefaniak

    2014-01-01

    Full Text Available Studies were made on the morphogenetic potential of petioles of Pelargonium hortorum (1 variety, 2 clones and Pelargonium peltatum (1 variety, 1 clone grown on the medium of Murashige and Skoog (MS supplemented with auxins (IAA, IBA, NAA and cytokinins (KIN, BAP, zeatin. The most intensive growth of callus of both species was observed on medium supplemented with 0.1 mg/l NAA and 10 mg/l KIN, both under constant illumination and in darkness. Differentiation of callus and regeneration of plants occurred in P. peltatum variety "PAC Dresdner Amethyst" and in P. hortorum clone 3766/4 on medium containing 1 mg/l BAP, 1 mg/l IAA and exposed to a photoperiod (L:D = 16:8.

  7. Establishment of Cell Suspension Culture and Plant Regeneration in Abrus precatorius L., a Rare Medicinal Plant

    Directory of Open Access Journals (Sweden)

    Mohammad Serajur RAHMAN

    2012-02-01

    Full Text Available A new protocol has been developed for cell culture and in vitro regeneration of Abrus precatorius that holds enormous potentiality for preparation of medicines. In vitro grown calli were cultured in Murashige and Skoog (MS liquid media in agitated condition fortified with 0.5 mg/l 6-Benzylaminopurine. Growth curve of cells revealed that the cells continued to grow until 12 days of culture and got the highest peak from day 6-8. Isolated cell was found to produce highest 8.2% calli when suspended on MS medium supplemented with 0.5 mg/l 6-Benzylaminopurine and 0.1 mg/l 1-Naphthaleneacetic acid. Callus derived from single cell produced highest number of embryo (25-28% cultured on MS medium fortified with 2.0 mg/l 6-Benzylaminopurine and 0.2 mg/l 1-Naphthaleneacetic acid. The bipolar embryos were selected and optimum shoot formation was recorded on MS medium supplemented with 2.0 mg/l 6-Benzylaminopurine and 0.1 mg/l 1-Naphthaleneacetic acid. The optimum root induction was noticed in MS medium supplemented with 1.0 mg/l 3-Indolebutyric acid. Rooted plantlets were successfully transferred to potting soil and acclimatized to outdoor conditions.

  8. Protoplast isolation and genetically true-to-type plant regeneration from leaf- and callus-derived protoplasts of Albizia julibrissin

    Science.gov (United States)

    Mohammad-Shafie Rahmani; Paula M. Pijut; Naghi Shabanian

    2016-01-01

    Protoplast isolation and subsequent plant regeneration of Albizia julibrissin was achieved from leaf and callus explants. Leaf tissue from 4 to 5-week-old in vitro seedlings was the best source for high-yield protoplast isolation. This approach produced 7.77 × 105 protoplasts (Pp) per gram fresh weight with 94 % viability;...

  9. Genetic fidelity assessment of in vitro-regenerated plants of Albizia julibrissin using SCoT and IRAP fingerprinting

    Science.gov (United States)

    Mohammad-Shafie Rahmani; Paula M. Pijut; Naghi Shabanian; Mona. Nasri

    2015-01-01

    A protocol was established for callus induction and plant regeneration of Albizia julibrissin Durazz., a multipurpose tree. Calli were induced on hypocotyl explants excised from 10- to 14-d-old in vitro seedlings cultured on Murashige and Skoog (MS) medium supplemented with α-naphthaleneacetic acid (NAA) alone or in...

  10. High frequency somatic embryogenesis and plant regeneration from zygotic embryo-derived callus cultures of three Allium species

    NARCIS (Netherlands)

    Valk, P. van der; Scholten, O.E.; Verstappen, F.; Jansen, R.C.; Dons, J.J.M.

    1992-01-01

    The plant regeneration ability of zygotic embryo-derived callus cultures was studied for 12 A. cepa varieties and accessions, two A. fistulosum varieties, one A. fistulosum × A. cepa interspecific hybrid and two A. porrum varieties. Compact embryogenic callus was induced on Murashige and Skoog (MS)

  11. Protoplast isolation and plant regeneration of guava (Psidium guajava L.) using experiments in mixture-amount design

    Science.gov (United States)

    A protocol was established for plant regeneration from leaf protoplasts of guava (Psidium guajava L.) using mixture-amount (concentration) experiments. A protoplast yield of 3.7 × 106 (viability > 90 percent) was obtained when 1 g leaf strips were digested in a solution of approximately 0.75 M osmot...

  12. Regeneration of whole fertile plants from 30,000-y-old fruit tissue buried in Siberian permafrost.

    Science.gov (United States)

    Yashina, Svetlana; Gubin, Stanislav; Maksimovich, Stanislav; Yashina, Alexandra; Gakhova, Edith; Gilichinsky, David

    2012-03-06

    Whole, fertile plants of Silene stenophylla Ledeb. (Caryophyllaceae) have been uniquely regenerated from maternal, immature fruit tissue of Late Pleistocene age using in vitro tissue culture and clonal micropropagation. The fruits were excavated in northeastern Siberia from fossil squirrel burrows buried at a depth of 38 m in undisturbed and never thawed Late Pleistocene permafrost sediments with a temperature of -7 °C. Accelerator mass spectrometry (AMS) radiocarbon dating showed fruits to be 31,800 ± 300 y old. The total γ-radiation dose accumulated by the fruits during this time was calculated as 0.07 kGy; this is the maximal reported dose after which tissues remain viable and seeds still germinate. Regenerated plants were brought to flowering and fruiting and they set viable seeds. At present, plants of S. stenophylla are the most ancient, viable, multicellular, living organisms. Morphophysiological studies comparing regenerated and extant plants obtained from modern seeds of the same species in the same region revealed that they were distinct phenotypes of S. stenophylla. The first generation cultivated from seeds obtained from regenerated plants progressed through all developmental stages and had the same morphological features as parent plants. The investigation showed high cryoresistance of plant placental tissue in permafrost. This natural cryopreservation of plant tissue over many thousands of years demonstrates a role for permafrost as a depository for an ancient gene pool, i.e., preexisting life, which hypothetically has long since vanished from the earth's surface, a potential source of ancient germplasm, and a laboratory for the study of rates of microevolution.

  13. Response of planted northern red oak seedlings to regeneration harvesting, Midstory removal, and prescribed burning

    Science.gov (United States)

    Stacy L. Clark; Scott E. Schlarbaum; Tara L. Keyser; Callie J. Schweitzer; Marty Spetich; Dean Simon; Gordon S. Warburton

    2016-01-01

    Oak (Quercus) is difficult to naturally regenerate in many mature oak stands on productive sites in the southeastern United States, and artificial regeneration alternatives should be considered. Artificial regeneration can potentially restore or enrich the oak component at the stand level. We examined genetic and silvicultural effects on...

  14. Rapid divergence of ecotypes of an invasive plant

    Science.gov (United States)

    Ray, Avik; Ray, Rajasri

    2014-01-01

    Invasive species demonstrate rapid evolution within a very short period of time allowing one to understand the underlying mechanism(s). Lantana camara, a highly invasive plant of the tropics and subtropics, has expanded its range and successfully established itself almost throughout India. In order to uncover the processes governing the invasion dynamics, 218 individuals from various locations across India were characterized with six microsatellites. By integrating genetic data with niche modelling, we examined the effect of drift and environmental selection on genetic divergence. We found multiple genetic clusters that were non-randomly distributed across space. Spatial autocorrelation revealed a strong fine-scale structure, i.e. isolation by distance. In addition, we obtained evidence of inhibitory effects of selection on gene flow, i.e. isolation by environmental distance. Perhaps, local adaptation in response to selection is offsetting gene flow and causing the populations to diverge. Niche models suggested that temperature and precipitation play a major role in the observed spatial distribution of this plant. Based on a non-random distribution of clusters, unequal gene flow among them and different bioclimatic niche requirements, we concluded that the emergence of ecotypes represented by two genetic clusters is underway. They may be locally adapted to specific climatic conditions, and perhaps at the very early stages of ecological divergence. PMID:25165061

  15. Automated regenerable microarray-based immunoassay for rapid parallel quantification of mycotoxins in cereals.

    Science.gov (United States)

    Oswald, S; Karsunke, X Y Z; Dietrich, R; Märtlbauer, E; Niessner, R; Knopp, D

    2013-08-01

    An automated flow-through multi-mycotoxin immunoassay using the stand-alone Munich Chip Reader 3 platform and reusable biochips was developed and evaluated. This technology combines a unique microarray, prepared by covalent immobilization of target analytes or derivatives on diamino-poly(ethylene glycol) functionalized glass slides, with a dedicated chemiluminescence readout by a CCD camera. In a first stage, we aimed for the parallel detection of aflatoxins, ochratoxin A, deoxynivalenol, and fumonisins in cereal samples in a competitive indirect immunoassay format. The method combines sample extraction with methanol/water (80:20, v/v), extract filtration and dilution, and immunodetection using horseradish peroxidase-labeled anti-mouse IgG antibodies. The total analysis time, including extraction, extract dilution, measurement, and surface regeneration, was 19 min. The prepared microarray chip was reusable for at least 50 times. Oat extract revealed itself as a representative sample matrix for preparation of mycotoxin standards and determination of different types of cereals such as oat, wheat, rye, and maize polenta at relevant concentrations according to the European Commission regulation. The recovery rates of fortified samples in different matrices, with 55-80 and 58-79%, were lower for the better water-soluble fumonisin B1 and deoxynivalenol and with 127-132 and 82-120% higher for the more unpolar aflatoxins and ochratoxin A, respectively. Finally, the results of wheat samples which were naturally contaminated with deoxynivalenol were critically compared in an interlaboratory comparison with data obtained from microtiter plate ELISA, aokinmycontrol® method, and liquid chromatography-mass spectrometry and found to be in good agreement.

  16. In vitro plant regeneration of Albizia lebbeck (L. from seed explants

    Directory of Open Access Journals (Sweden)

    S. Perveen

    2013-07-01

    Full Text Available Objectives: An efficient and reproducible regeneration protocol for rapid multiplication of Albizia lebbeck (L. was developed by using intact seed explants.Methods: Murashige and Skoog's (MS medium supplemented with different hormones (BA, Kn, GA3 and TDZ was used for the induction of multiple shoots from the seed explants. Ex-vitro rooting was performed by using pulse treatment method in auxins (IBA and NAA and the complete plantlets were transferred to the field.Results: High frequency direct shoot induction was found in aseptic seed cultures of A. lebbeck on Murashige and Skoog medium supplemented with 5.0 µM TDZ (Thiadiazuron. Seeds were germinated after 7 days of culture and induced maximum 8 shoots from the region adjacent to the apex of the primary shoot of the seedling upto 25 days of incubation. Proliferating shoot cultures with increased shoot length was established by sub-culture of excised sprouting epicotyls on MS medium supplied with reduced concentrations of TDZ. Maximum shoot regeneration frequency (76 % with  highest number of shoots (21 and shoot length (5.1 cm per sprouting epicotyl was observed in the MS medium supplemented with 0.5 µM TDZ after 8 weeks of culture. Different concentrations of Indole-3-butyric acid (IBA and α-naphthalene acetic acid (NAA were tested to determine the optimal conditions for ex-vitro rooting of the microshoots. The best treatment for maximum ex-vitro root induction frequency (81 % was accomplished with IBA (250 µM pulse treatment given to the basal end of the microshoots for 30 min followed by their transfer in plastic cups containing soilrite and eventually established in normal garden soil + soilrite (1:1 with 78 % survival rate. In addition, histological study was undertaken to gain a better understanding of the regenerated shoots from the epicotyl region.Conclusion: The findings will be fruitful in getting a time saving and cost effective protocol for the in vitro propagation of Albizia

  17. Doubled haploid production from Spanish onion (Allium cepa L.) germplasm: embryogenesis induction, plant regeneration and chromosome doubling

    Science.gov (United States)

    Fayos, Oreto; Vallés, María P.; Garcés-Claver, Ana; Mallor, Cristina; Castillo, Ana M.

    2015-01-01

    The use of doubled haploids in onion breeding is limited due to the low gynogenesis efficiency of this species. Gynogenesis capacity from Spanish germplasm, including the sweet cultivar Fuentes de Ebro, the highly pungent landrace BGHZ1354 and the two Valenciana type commercial varieties Recas and Rita, was evaluated and optimized in this study. The OH-1 population, characterized by a high gynogenesis induction, was used as control. Growing conditions of the donor plants were tested with a one-step protocol and field plants produced a slightly higher percentage of embryogenesis induction than growth chamber plants. A one-step protocol was compared with a two-step protocol for embryogenesis induction. Spanish germplasm produced a 2–3 times higher percentage of embryogenesis with the two-step protocol, Recas showing the highest percentage (2.09%) and Fuentes de Ebro the lowest (0.53%). These percentages were significantly lower than those from the OH-1 population, with an average of 15% independently of the protocol used. The effect of different containers on plant regeneration was tested using both protocols. The highest percentage of acclimated plants was obtained with the two-step protocol in combination with Eco2box (70%), whereas the lowest percentage was observed with glass tubes in the two protocols (20–23%). Different amiprofos-methyl (APM) treatments were applied to embryos for chromosome doubling. A similar number of doubled haploid plants were recovered with 25 or 50 μM APM in liquid medium. However, the application of 25 μM in solid medium for 24 h produced the highest number of doubled haploid plants. Somatic regeneration from flower buds of haploid and mixoploid plants proved to be a successful approach for chromosome doubling, since diploid plants were obtained from the four regenerated lines. In this study, doubled haploid plants were produced from the four Spanish cultivars, however further improvements are needed to increase their gynogenesis

  18. In vitro regeneration of Drosera burmannii Vahl.: a carnivorous plant of north-east India.

    Science.gov (United States)

    Yanthan, J Sureni; Kehie, Mechuselie; Kumaria, Suman; Tandon, Pramod

    2017-06-01

    An efficient in vitro regeneration protocol has been developed from shoot tips of Drosera burmannii Vahl., a carnivorous plant of north-east India. Various plant growth regulators were used to study their efficacy in the induction of multiple shoots and roots. Of the various treatments, the maximum number of shoots (28.8 ± 1.5) and roots (9.7 ± 0.6) was observed in one-fourth strength standard medium (MS with 50 mg/l citric acid and 10 mg/l ascorbic acid) supplemented with 4 mg/l 6-benzylaminopurine (BAP) and 4 mg/l α-naphthalene acetic acid (NAA) followed by 26.8 ± 1.4 shoots in one-fourth strength SM fortified with 4 mg/l kinetin (KN) and 4 mg/l NAA. The well-developed plantlets with shoots and roots were potted in small plastic glasses filled with a mixture of sand and farmyard manure (3:1); these plantlets when transferred to a glasshouse for hardening and acclimatization showed 90% survival.

  19. Biological activity of the tzs gene of nopaline Agrobacterium tumefaciens GV3101 in plant regeneration and genetic transformation.

    Science.gov (United States)

    Han, Zhao-Fen; Hunter, David M; Sibbald, Susan; Zhang, Ji-Shu; Tian, Lining

    2013-11-01

    Agrobacterium tumefaciens has been widely used in plant genetic transformation. Hormone-encoding genes residing in the T-DNA region have been removed, resulting in disarmed Agrobacterium strains that are used in various transformation experiments. Nopaline Agrobacterium strains, however, carry another hormone gene, trans-zeatin synthesizing (tzs), that codes for trans-zeatin in the virulence region of the tumor-inducing plasmids. We investigated the activity and function of the tzs gene of a nopaline Agrobacterium sp. strain GV3101 in plant in vitro regeneration. Leaf explants of tobacco and Nicotiana benthamiana co-cultured with strain GV3101 exhibited active shoot regeneration in media without added plant growth regulators. On medium without plant growth regulators, transgenic shoots were also induced from explants co-cultured with GV3101 containing a binary vector. Enzyme-linked immunosorbent assay showed that cell-free extracts of Agrobacterium sp. strain GV3101 culture contained the trans-zeatin at 860 ng/liter. Polymerase chain reaction using tzs-specific primers showed that the tzs gene was present in strain GV3101 but not in other Agrobacterium strains. The study showed that the tzs gene in GV3101 was actively expressed, and that trans-zeatin produced in the Agrobacterium strain can promote plant shoot regeneration.

  20. In vitro plant regeneration of two cucumber (Cucumis sativum L. genotypes: Effects of explant types and culture medium

    Directory of Open Access Journals (Sweden)

    Grozeva Stanislava

    2014-01-01

    Full Text Available The effect of different phytohormone concentrations on callusogenesis and organogenesis in two cucumber genotypes were studied. It was established that the rate of plant regeneration depends on genotype, explant type and culture medium. Hypocotyls were found to be more responsive than cotyledons in morphogenesis. In vitro planlet-regenerants have been obtained in hypocotyls explants on culture medium with 1.0 and 2.0 mgL-1 BA for cultivar Gergana and in 1.0 and 3.0 mgL-1K-line 15B. Induction of regeneration in cotyledons were established only in cultivar Gergana on culture medium supplemented with 3.0 mgL-1 BA and in combination of 0.5 mgL-1IAA.

  1. A novel combination of plant growth regulators for in vitro regeneration of complete plantlets of guar [Cyamopsis tetragonoloba (L.) Taub].

    Science.gov (United States)

    Verma, S; Gill, K S; Pruthi, V; Dhugga, K S; Randhaw, G S

    2013-12-01

    A novel combination of plant growth regulators comprising indole-3-butyric acid (IBA), 6-benzylaminopurine (BA) and gibberellic acid (GA3) in Murashige and Skoog basal medium has been formulated for in vitro induction of both shoot and root in one culture using cotyledonary node explants of guar, (Cyamopsis tetragonoloba). Highest percentages of shoot (92%) and root (80%) induction were obtained in the medium containing (mg/L) 2 IBA, 3 BA and 1 GA3. Shoot regeneration from the cotyledonary node explants was observed after 10-15 days. Regeneration of roots from these shoots occurred after 20 to 25 days. The regenerated plantlets showed successful acclimatization on transfer to soil. This protocol is expected to be helpful in carrying out various in vitro manipulations in this economically and industrially important legume.

  2. The regeneration of epidermal cells of Saintpaulia leaves as a new plant-tissue system for cellular radiation biology.

    Science.gov (United States)

    Engels, F M; van der Laan, F M; Leenhouts, H P; Chadwick, K H

    1980-09-01

    Investigation of the nucleus of epidermal cells of the petioles of Saintpaulia leaves by cytofluorimetry revealed that all cells are in a non-cycling pre DNA synthesis phase. Cultivation of dissected leaves results in a synchronous regeneration process of a defined number of cells. Five days after onset of cultivation the cells reach the first mitosis. The nuclear development during the regeneration process is described. Irradiation of the leaves results in a directly visible inhibition of this regenerating capability which is used to quantify cell survival in a tissue. The data show that the radiation response has a similar shape to that of the survival of single cells in culture. This response can be observed before the first mitosis of the cells and its application as a new plant tissue system for cellular radiation research is discussed.

  3. Plant regeneration from cotyledonary explants of Eucalyptus camaldulensis dehn and histological study of organogenesis in vitro

    Directory of Open Access Journals (Sweden)

    Roberson Dibax

    2010-04-01

    Full Text Available The present work aimed at regenerating plants of Eucalyptus camaldulensis from the cotyledonary explants and describing the anatomy of the tissues during callogenesis and organogenesis processes, in order to determine the origin of the buds. The cotyledonary leaves of E. camaldulensis were cultured in Murashige and Skoog (MS, WPM and JADS media supplemented with 2.7 µM NAA and 4.44 µM BAP. The best results for bud regeneration were obtained on MS and WPM media (57.5 and 55% of calluses formed buds, respectively. Shoot elongation and rooting (80% were obtained on MS/2 medium (with half-strength salt concentration with 0.2% activated charcoal. Acclimatization was performed in the growth chamber for 48 h and then the plants were transferred to a soil:vermiculite mixture and cultured in a greenhouse. Histological studies revealed that the callogenesis initiated in palisade parenchyma cells and that the adventitious buds were formed from the calluses, indicating indirect organogenesis.Este trabalho teve como objetivo a obtenção de plantas de Eucalyptus camaldulensis a partir de folhas cotiledonares e o estudo da anatomia dos tecidos durante a calogênese e organogênese para determinar a origem das gemas. Folhas cotiledonares foram cultivadas em meios de cultura MS, WPM e JADS suplementados com 2,7 µM de ANA e 4,44 µM de BAP. Os melhores resultados para a regeneração de gemas foram obtidos com os meios MS e WPM. Para o alongamento e enraizamento, o meio de cultura MS/2 contendo 0,2% de carvão ativado apresentou-se eficiente para ambas as etapas. A aclimatização foi realizada mediante a abertura dos frascos na sala de crescimento por 48 horas, seguido da transferência para casa-de-vegetação com nebulização intermitente. Estudos histológicos foram conduzidos e revelaram que a calogênese teve início nas células do parênquima paliçádico e que as gemas adventícias formaram-se a partir dos calos, indicando a organogênese indireta.

  4. Embryogenesis induction, callogenesis, and plant regeneration by in vitro culture of tomato isolated microspores and whole anthers.

    Science.gov (United States)

    Seguí-Simarro, José M; Nuez, Fernando

    2007-01-01

    In this work, some of the different in vitro developmental pathways into which tomato microspores or microsporocytes can be deviated experimentally were explored. The two principal ones are direct embryogenesis from isolated microspores and callus formation from meiocyte-containing anthers. By means of light and electron microscopy, the process of early embryogenesis from isolated microspores and the disruption of normal meiotic development and change of developmental fate towards callus proliferation, morphogenesis, and plant regeneration have been shown. From microspores isolated at the vacuolate stage, embryos can be directly induced, thus avoiding non-androgenic products. In contrast, several different morphogenic events can be triggered in cultures of microsporocyte-containing anthers under adequate conditions, including indirect embryogenesis, adventitious organogenesis, and plant regeneration. Both callus and regenerated plants may be haploid, diploid, and mostly mixoploid. The results demonstrate that both gametophytic and sporophytic calli occur in cultured tomato anthers, and point to an in vitro-induced disturbance of cytokinesis and subsequent fusion of daughter nuclei as a putative cause for mixoploidy and genome doubling during both tetrad compartmentalization and callus proliferation. The potential implications of the different alternative pathways are discussed in the context of their application to the production of doubled-haploid plants in tomato, which is still very poorly developed.

  5. AtTCTP2, an Arabidopsis thaliana homolog of Translationally Controlled Tumor Protein, enhances in vitro plant regeneration

    Science.gov (United States)

    Toscano-Morales, Roberto; Xoconostle-Cázares, Beatriz; Cabrera-Ponce, José L.; Hinojosa-Moya, Jesús; Ruiz-Salas, Jorge L.; Galván-Gordillo, Santiago V.; Guevara-González, Ramón G.; Ruiz-Medrano, Roberto

    2015-01-01

    The Translationally Controlled Tumor Protein (TCTP) is a central regulator of cell proliferation and differentiation in animals, and probably also in plants. Arabidopsis harbors two TCTP genes, AtTCTP1 (At3g16640), which is an important mitotic regulator, and AtTCTP2 (At3g05540), which is considered a pseudogene. Nevertheless, we have obtained evidence suggesting that this gene is functional. Indeed, a T-DNA insertion mutant, SALK_045146, displays a lethal phenotype during early rosette stage. Also, both the AtTCTP2 promoter and structural gene are functional, and heterozygous plants show delayed development. AtTCTP1 cannot compensate for the loss of AtTCTP2, since the accumulation levels of the AtTCTP1 transcript are even higher in heterozygous plants than in wild-type plants. Leaf explants transformed with Agrobacterium rhizogenes harboring AtTCTP2, but not AtTCTP1, led to whole plant regeneration with a high frequency. Insertion of a sequence present in AtTCTP1 but absent in AtTCTP2 demonstrates that it suppresses the capacity for plant regeneration; also, this phenomenon is enhanced by the presence of TCTP (AtTCTP1 or 2) in the nuclei of root cells. This confirms that AtTCTP2 is not a pseudogene and suggests the involvement of certain TCTP isoforms in vegetative reproduction in some plant species. PMID:26191065

  6. AtTCTP2, an Arabidopsis thaliana homolog of Translationally Controlled Tumor Protein, enhances in vitro plant regeneration

    Directory of Open Access Journals (Sweden)

    Roberto eToscano-Morales

    2015-07-01

    Full Text Available The Translationally Controlled Tumor Protein (TCTP is a central regulator of cell proliferation and differentiation in animals, and probably also in plants. Arabidopsis harbors two TCTP genes, AtTCTP1 (At3g16640, which is an important mitotic regulator, and AtTCTP2 (At3g05540, which is considered a pseudogene. Nevertheless, we have obtained evidence suggesting that this gene is functional. Indeed, a T-DNA insertion mutant, SALK_045146, displays a lethal phenotype during early rosette stage. Also, both the AtTCTP2 promoter and structural gene are functional, and heterozygous plants show delayed development. AtTCTP1 cannot compensate for the loss of AtTCTP2, since the accumulation levels of the AtTCTP1 transcript are even higher in heterozygous plants than in wild-type plants. Leaf explants transformed with Agrobacterium rhizogenes harboring AtTCTP2, but not AtTCTP1, led to whole plant regeneration with a high frequency. Insertion of a sequence present in AtTCTP1 but absent in AtTCP2 demonstrates that this suppresses the capacity for plant regeneration; also, this phenomenon requires the presence of TCTP (AtTCTP1 or 2 in the nuclei of root cells. This confirms that AtTCTP2 is not a pseudogene and suggests the involvement of certain TCTP isoforms in vegetative reproduction in some plant species.

  7. Establishment of an Efficient In Vitro Regeneration Protocol for Rapid and Mass Propagation of Dendrobium chrysotoxum Lindl. Using Seed Culture

    Science.gov (United States)

    2014-01-01

    An efficient in vitro regeneration protocol from seed culture has been established successfully for Dendrobium chrysotoxum, an epiphytic orchid having tremendous ornamental and medicinal values. Seed germination response was encouraging in Mitra (M) medium enriched with different combinations of auxins and cytokinins. Medium supplemented with 0.4% activated charcoal (AC), 2 mg/L 6-benzyl amino purine (BAP), and 2 mg/L indole-3-acetic acid (IAA) produced best seed germination percentage in 2 weeks of culture. Incorporation of higher concentration of kinetin (KN) or BAP in combination with low auxin in medium induced pronounced shooting and leaf formation. Reduction in leaf development was evident when cytokinins exist singly in medium indicating synergistic effect of auxin and cytokinin in leaf induction. Presence of elevated level of indole-3-butyric acid (IBA) or 1-naphthalene acetic acid (NAA) with low cytokinin content in medium generated more in vitro rooting, though IBA was found to be more effective in rooting induction as compared to NAA. The in vitro protocol for asymbiotic seed germination developed from the present investigation can be used for rapid mass propagation of this highly important Dendrobium orchid species. PMID:25401154

  8. Establishment of an efficient in vitro regeneration protocol for rapid and mass propagation of Dendrobium chrysotoxum Lindl. using seed culture.

    Science.gov (United States)

    Nongdam, Potshangbam; Tikendra, Leimapokpam

    2014-01-01

    An efficient in vitro regeneration protocol from seed culture has been established successfully for Dendrobium chrysotoxum, an epiphytic orchid having tremendous ornamental and medicinal values. Seed germination response was encouraging in Mitra (M) medium enriched with different combinations of auxins and cytokinins. Medium supplemented with 0.4% activated charcoal (AC), 2 mg/L 6-benzyl amino purine (BAP), and 2 mg/L indole-3-acetic acid (IAA) produced best seed germination percentage in 2 weeks of culture. Incorporation of higher concentration of kinetin (KN) or BAP in combination with low auxin in medium induced pronounced shooting and leaf formation. Reduction in leaf development was evident when cytokinins exist singly in medium indicating synergistic effect of auxin and cytokinin in leaf induction. Presence of elevated level of indole-3-butyric acid (IBA) or 1-naphthalene acetic acid (NAA) with low cytokinin content in medium generated more in vitro rooting, though IBA was found to be more effective in rooting induction as compared to NAA. The in vitro protocol for asymbiotic seed germination developed from the present investigation can be used for rapid mass propagation of this highly important Dendrobium orchid species.

  9. Efficient in vitro plant regeneration from immature zygotic embryos of pearl millet [Pennisetum glaucum (L.) R. Br.] and Sorghum bicolor (L.) Moench

    CSIR Research Space (South Africa)

    Oldach, KH

    2001-07-01

    Full Text Available to regeneration medium R or L3 (Table 1) and cultured under light conditions (70?5 ?E/m2/s1) for 16 h per day. Rooting was supported on half-strength, hormone-free L3 medium contain- ing 1.5% sucrose. Subculturing was carried out weekly on callus induction medium...?E, to medium R or to hormone-free L3 medium (Table 2). Regeneration rates were evaluated after 10 weeks of cul- ture. Nearly no plants regenerated when embryos were induced on media I?P, containing 0.5 and 1 mg/l cytoki- nin. The highest numbers of regenerated...

  10. Regeneration of Cytologically Stable Plants Through Dedifferentiation, Redifferentiation, and Artificial Seeds in Spathoglottis plicata Blume. (Orchidaceae

    Directory of Open Access Journals (Sweden)

    Sk Moquammel Haque

    2017-09-01

    Full Text Available Spathoglottis plicata Blume. is a horticulturally important vulnerable ground orchid with beautiful flowers blooming round the year. High-frequency protocorm-like body (PLB formation was established via callus culture from vegetative tissues of in vitro germinated seedlings of S. plicata. Media containing MS salts and Gamborg's B5 vitamins supplemented with 1.0 mg⋅L−1 2,4-dichlorophenoxyacetic acid (2,4-D, 3.0 mg⋅L−1 α-naphthaleneacetic acid (NAA, 1.0 mg⋅L−1 kinetin (KIN, and 10% (v/v ‘Aloe vera gel’ (AvG were effective in fragile calli induction. A maximum of (22.3 ± 0.52 PLBs were induced from about 250 mg callus within 45–55 days in the presence of 2.0 mg⋅L−1 NAA and 3.0 mg⋅L−1 6-benzylaminopurine (BAP. Briefly, 3.0% sodium alginate was found to be most suitable for the formation of an appropriate shape and good germination rates (86.7% of artificial seeds. Out of three different temperatures (4, 15, and 24 °C, the best result was achieved at 4 °C with 66.7% germinability even after 90 days of storage. Plantlets were acclimatized with 86.6% survival rate and 76.3% of these plants produced flowers within 12–15 months of field transfer. Chromosomal studies revealed cytological stability of all regenerants containing 2n = 40 chromosomes as in the parental plants. The present protocol can be applied reliably for the purposes of large-scale commercial propagation and short-term conservation of this orchid.

  11. Direct regeneration of Periwinkle (Catharanthus roseus) via node explants culture and different combinations of plant growth regulators

    OpenAIRE

    M. Talebi; F. Etesam; B.E. Sayed-Tabatabaei; Gh. Khaksar

    2012-01-01

    Periwinkle (Catharanthus roseus L., Apocynaceae) contains more than 130 different terpenoid indole alkaloids (TIAs), of which two dimeric alkaloids, Vinblastine and Vincristine, have antineoplastic activity and are useful in treatment of various cancers. Specific production of some alkaloids in differentiated tissues such as leaf and stem led to use direct regeneration of explants in order to increase the production of these important alkaloids in the plant. In this research, 30 combinations ...

  12. Woody plant regeneration after blowdown, salvage logging, and prescribed fire in a northern Minnesota forest

    Science.gov (United States)

    Brian J. Palik; Doug. Kastendick

    2009-01-01

    Salvage logging after natural disturbance has received increased scrutiny in recent years because of concerns over detrimental effects on tree regeneration and increased fine fuel levels. Most research on tree regeneration after salvage logging comes from fire-prone systems and is short-term in scope. Limited information is available on longer term responses to salvage...

  13. Plant regeneration, genetic fidelity, and active ingredient content of encapsulated hairy roots of Picrorhiza kurrooa Royle ex Benth.

    Science.gov (United States)

    Rawat, Janhvi Mishra; Rawat, Balwant; Mehrotra, Shakti

    2013-06-01

    Among five hairy root lines of Picrorhiza kurrooa that were established through Agrobacterium rhizogenes, one (H7) was selected for encapsulation due to high accumulation of picrotin and picrotoxinin (8.3 and 47.6 μg/g DW, respectively). Re-grown encapsulated roots induced adventitious shoots with 73 % frequency on MS medium supplemented with 0.1 μM 6-benzylaminopurine, following 6 months of storage at 25 °C. Regenerated plantlets had 85 % survival after 2 months. Regenerants were of similar morphotype having increased leaf number and branched root system as compared to non-transformed plants. The transformed nature of the plants was confirmed through PCR and Southern blot analysis. Genetic fidelity analysis of transformed plants using RAPD and ISSR showed 5.2 and 3.6 % polymorphism, respectively. Phytochemical analysis also showed that picrotin and picrotoxinin content were similar in hairy root line and its regenerants.

  14. In vitro plant regeneration system for tropical butternut squash genotypes (Cucurbita moschata

    Directory of Open Access Journals (Sweden)

    Marta Valdez-Melara

    2009-11-01

    Full Text Available An efficient and reproducible method for regeneration of commercial and pure lines of tropical butternut squash (Cucurbita moschata plants via somatic embryogenesis was developed. The influence of genotype, explant source, N6-benzylaminopurine (BAP, 2,4-dichlorophenoxyacetic acid (2,4-D and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T concentration on somatic embryogenesis induction was investigated. Friable embryogenic calli was produced from zigotic embryos (53-56% and cotyledons from seedlings (70% of C. moschata cv. Sello de Oro cultured on callus induction medium (CIM supplemented with 0.5 mg/l or 3.5 mg/l 2,4-D. No embryogenic calli was obtained from leaf segments of C. moschata cv. Sello de Oro cultured on CIM supplemented with different concentrations of BAP and 2,4-D and cotyledons from seedlings of C. moschata cv. PVG 04 cultured on CIM with BAP and 2,4,5-T. Embryogenic calli induction was achieved in 75% C. moschata pure lines evaluated and calli percentage frequency range from 5% to 34%. Successful acclimatization of squash in vitro plants was achieved in the greenhouse and in the field. Regenerated plants appeared morphologically normal and set flowers and fruits with seeds that could germinate normally. Rev. Biol. Trop. 57 (Suppl. 1: 119-127. Epub 2009 November 30.En este estudio se desarrolló un método eficiente y reproducible para la regeneración de líneas puras de la planta tropical Cucurbita moschata mediante la vía de embriogénesis somática. Además se investigó acerca de la influencia del genotipo, transplante, y la concentración de N6-benzylaminopurina (BAP, 2,4-diclorofenoxyacetico ácido (2,4-D y 2,4,5-triclorofenoxyacetico ácido (2,4,5-T en la inducción de embriogénesis somática. Los callos embriogenéticos viables fueron producidos de embriones zigóticos (53-56% y cotiledones de semillas (70% de C. moschata cv. Sello de Oro cultivados en un medio de inducción de callos (CIM suplementado con 0.5 mg/l o 3.5 mg

  15. Regeneration of begonia plantlets by direct organogenesis

    OpenAIRE

    Mendi, Yalçın Y.; Mendi, Yalcin Y.; Curuk, P.; Kocaman, E.; Unek, C.; Eldoğan, S.; Eldogan, S.; Gencel, G.; Çetiner, Selim; Cetiner, Selim

    2009-01-01

    The economic importance of ornamentals worldwide suggests a bright future for ornamental breeding. Rapid progress in plant molecular biology has great potentials to contribute to the breeding of novel ornamental plants utilizing recombinant DNA technology. The plant cell, tissue or organ culture of many ornamental species and their regeneration are essential for providing the material and systems for their genetic manipulation, and this is therefore the first requirement of genetic engineerin...

  16. Effects of invasive alien kahili ginger (Hedychium gardnerianum) on native plant species regeneration in a Hawaiian rainforest

    Science.gov (United States)

    Minden, V.; Jacobi, J.D.; Porembski, S.; Boehmer, H.J.

    2010-01-01

    Questions: Does the invasive alien Hedychium gardnerianum (1) replace native understory species, (2) suppress natural regeneration of native plant species, (3) increase the invasiveness of other non-native plants and (4) are native forests are able to recover after removal of H. gardnerianum. Location: A mature rainforest in Hawai'i Volcanoes National Park on the island of Hawai'i (about 1200 m. a.s.l.; precipitation approximately 2770mm yr-1). Study sites included natural plots without effects of alien plants, ginger plots with a H. gardnerianum-domimted herb layer and cleared plots treated with herbicide to remove alien plants. Methods: Counting mature trees, saplings and seedlings of native and alien plant species. Using nonparametric H-tests to compare impact of H. gardnerianum on the structure of different sites. Results: Results confirmed the hypothesis that H. gardnerianum has negative effects on natural forest dynamics. Lower numbers of native tree seedlings and saplings were found on ginger-dominated plots. Furthermore, H. gardnerianum did not show negative effects on the invasive alien tree species Psidium cattleianum. Conclusions: This study reveals that where dominance of H. gardnerianum persists, regeneration of the forest by native species will be inhibited. Furthermore, these areas might experience invasion by P. cattleianum, resulting in displacement of native canopy species in the future, leading to a change in forest structure and loss of other species dependent on natural rainforest, such as endemic birds. However, if H. gardnerianum is removed the native Hawaiian forest is likely to regenerate and regain its natural structure. ?? 2009 International Association for Vegetation Science.

  17. Rapid identification of the medicinal plant Taraxacum formosanum ...

    African Journals Online (AJOL)

    Administrator

    2011-06-06

    Jun 6, 2011 ... College J. 8: 35-46. Xue CY, Li DZ, Lu JM, Yang JB, Liu JQ (2006). Molecular authentication of the traditional Tibetan medical plant Swertia mussotii. Planta Med. 72: 721-726. Yuan CC (2001). Textual research of material medica Taraxacum mongolicum and varietal identification. Chinese Wild Plant Res.

  18. Loss of CMD2‐mediated resistance to cassava mosaic disease in plants regenerated through somatic embryogenesis

    Science.gov (United States)

    Chauhan, Raj Deepika; Wagaba, Henry; Moll, Theodore; Alicai, Titus; Miano, Douglas; Carrington, James C.; Taylor, Nigel J.

    2016-01-01

    Summary Cassava mosaic disease (CMD) and cassava brown streak disease (CBSD) are the two most important viral diseases affecting cassava production in Africa. Three sources of resistance are employed to combat CMD: polygenic recessive resistance, termed CMD1, the dominant monogenic type, named CMD2, and the recently characterized CMD3. The farmer‐preferred cultivar TME 204 carries inherent resistance to CMD mediated by CMD2, but is highly susceptible to CBSD. Selected plants of TME 204 produced for RNA interference (RNAi)‐mediated resistance to CBSD were regenerated via somatic embryogenesis and tested in confined field trials in East Africa. Although micropropagated, wild‐type TME 204 plants exhibited the expected levels of resistance, all plants regenerated via somatic embryogenesis were found to be highly susceptible to CMD. Glasshouse studies using infectious clones of East African cassava mosaic virus conclusively demonstrated that the process of somatic embryogenesis used to regenerate cassava caused the resulting plants to become susceptible to CMD. This phenomenon could be replicated in the two additional CMD2‐type varieties TME 3 and TME 7, but the CMD1‐type cultivar TMS 30572 and the CMD3‐type cultivar TMS 98/0505 maintained resistance to CMD after passage through somatic embryogenesis. Data are presented to define the specific tissue culture step at which the loss of CMD resistance occurs and to show that the loss of CMD2‐mediated resistance is maintained across vegetative generations. These findings reveal new aspects of the widely used technique of somatic embryogenesis, and the stability of field‐level resistance in CMD2‐type cultivars presently grown by farmers in East Africa, where CMD pressure is high. PMID:26662210

  19. Afforestation by natural regeneration or by tree planting: examples of opposite hydrological impacts evidenced by long-term field monitoring in the humid tropics

    Science.gov (United States)

    Lacombe, G.; Ribolzi, O.; de Rouw, A.; Pierret, A.; Latsachak, K.; Silvera, N.; Pham Dinh, R.; Orange, D.; Janeau, J.-L.; Soulileuth, B.; Robain, H.; Taccoen, A.; Sengphaathith, P.; Mouche, E.; Sengtaheuanghoung, O.; Tran Duc, T.; Valentin, C.

    2015-12-01

    The humid tropics are exposed to an unprecedented modernization of agriculture involving rapid and highly-mixed land-use changes with contrasted environmental impacts. Afforestation is often mentioned as an unambiguous solution for restoring ecosystem services and enhancing biodiversity. One consequence of afforestation is the alteration of streamflow variability controlling habitats, water resources and flood risks. We demonstrate that afforestation by tree planting or by natural forest regeneration can induce opposite hydrological changes. An observatory including long-term field measurements of fine-scale land-use mosaics and of hydro-meteorological variables has been operating in several headwater catchments in tropical Southeast Asia since 2001. The GR2M water balance model repeatedly calibrated over successive 1 year periods, and used in simulation mode with specific rainfall input, allowed the hydrological effect of land-use change to be isolated from that of rainfall variability in two of these catchments in Laos and Vietnam. Visual inspection of hydrographs, correlation analyses and trend detection tests allowed causality between land-use changes and changes in seasonal flows to be ascertained. In Laos, the combination of shifting cultivation system (alternation of rice and fallow) and the gradual increase of teak tree plantations replacing fallow, led to intricate flow patterns: pluri-annual flow cycles induced by the shifting system, on top of a gradual flow increase over years caused by the spread of the plantation. In Vietnam, the abandonment of continuously cropped areas mixed with patches of tree plantations led to the natural re-growth of forest communities followed by a gradual drop in streamflow. Soil infiltrability controlled by surface crusting is the predominant process explaining why two modes of afforestation (natural regeneration or planting) led to opposite changes in flow regime. Given that commercial tree plantations will continue to

  20. Chloroplast ultra structure, photosynthesis and enzyme activities in regenerated plants of Stevia rebaudiana (Bert.) Bertoni as influenced by copper sulphate in the medium.

    Science.gov (United States)

    Jain, Pourvi; Kachhwaha, Sumita; Kothari, S L

    2014-09-01

    Stevia rebaudiana (Bert.) Bertoni is an important medicinal plant used as noncaloric commercial sweetener. Plants regenerated with higher levels of copper sulphate in the medium exhibited enhanced activity of peroxidase and polyphenoloxidase (PPO) enzymes. Transmission electron microscopy (TEM) revealed increase in size and number of electron dense inclusions in the chloroplasts of plants regenerated at optimised level of copper sulphate (0.5 microM) in the medium. There was decrease in chlorogenic acid (CGA) content. Chl-a-fluorescence transient pattern (OJIP) showed that the photosynthesis process was more efficient at 0.5 microM CuSO4 in the medium.

  1. In vitro regeneration of Turkish dwarf chickling (Lathyrus cicera L ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-06-17

    Jun 17, 2008 ... in the developing animal feed market, it is necessary to investigate different approaches for rapid multiplication of the plant for future use in the breeding programmes. In vitro plant regeneration has been reported in L. cicera through cotyledonary node (Malik et al., 1992), L. sativus through cotyledonary ...

  2. Plant regeneration from hypocotyl protoplasts of winter oilseed rape (Brassica napus L.

    Directory of Open Access Journals (Sweden)

    Wacław Orczyk

    2014-01-01

    Full Text Available Protoplasts were isolated from hypocotyls of six breeding lines and two cultivars of winter oilseed rape (B. napus L.. Under presented culture conditions almost all of the protoplasts regenerated cell walls. Division frequency depended on the genotype and was from 50% to 64%. Shoot regeneration (also depended on the genotype was induced with the frequency of 3.6% (for cv Bolko on the medium containing IAA (0.1 mg•dm-3, zeatin (0.5 mg•dm-3 and BAP (0.5 mg•dm-3 . All shoots were rooted on MS basal medium supplemented with sucrose 30 g•dm-3.

  3. Rapid identification of the medicinal plant Taraxacum formosanum ...

    African Journals Online (AJOL)

    Administrator

    2011-06-06

    Jun 6, 2011 ... Original identification of medicinal plants is essential for quality control. In this study, the internal transcribed spacer 2 (ITS2) nuclear ribosomal DNA served as a DNA barcode and was amplified by allele-specific PCR. This approach was exploited to differentiate Taraxacum formosanum from five.

  4. Contrasting effects of fire severity on regeneration of the dominant woody species in two coastal plant communities at Wilsons Promontory, Victoria

    OpenAIRE

    Morgan, John W.; Nield, Catharine M.

    2013-01-01

    Following wildfire in 2005 at Wilsons Promontory, Victoria, we asked how fire severity affected the postfire regeneration of dominant woody species in two coastal plant communities. We documented the effects of fire severity (unburned, low, high) on stand mortality and seedling regeneration in shrublands dominated by the obligate seeder Leptospermum laevigatum (Myrtaceae) and woodlands dominated by the resprouting Banksia integrifolia var. integrifolia (Proteaceae). Leptospermum laevigatum is...

  5. An efficient method for Agrobacterium-mediated genetic transformation and plant regeneration in cumin (Cuminum cyminum L.).

    Science.gov (United States)

    Pandey, Sonika; Mishra, Avinash; Patel, Manish Kumar; Jha, Bhavanath

    2013-09-01

    Cumin is an annual herbaceous medicinally important plant having diverse applications. An efficient and reproducible method of Agrobacterium-mediated genetic transformation was herein established for the first time. A direct regeneration method without callus induction was optimised using embryos as explant material in Gamborg's B5 medium supplemented with 0.5-μM 6-benzyladenine and 2.0-μM α-naphthalene acetic acid. About 1,020 embryos (a mean of 255 embryos per batch) were used for the optimisation of transformation conditions. These conditions were an Agrobacterium cell suspension of 0.6 OD600, a co-cultivation time of 72 h, 300-μM acetosyringone and wounding of explants using a razor blade. Pre-cultured elongated embryos were treated using optimised conditions. About 720 embryos (a mean of 180 embryos per batch) were used for transformation and 95 % embryos showed transient β-glucuronidase expression after co-cultivation. Putative transformed embryos were cultured on B5 medium for shoot proliferation and 21 regenerated plants were obtained after selection and allowed to root. T0 plantlets showed β-glucuronidase expression and gene integration was confirmed via PCR amplification of 0.96 and 1.28 kb fragments of the hygromycin-phosphotransferase II and β-glucuronidase genes, respectively. In this study, a transformation efficiency of 1.5 % was demonstrated and a total of 11 transgenic plants were obtained at the hardening stage, however, only four plants acclimatised during hardening. Gene copy number was analysed by Southern blot analysis of hardened plants and single-copy gene integration was observed. This is the first successful attempt of Agrobacterium-mediated genetic transformation of cumin.

  6. Optimized growth and plant regeneration for callus of Lilium longiflorum cv. Nellie White

    Science.gov (United States)

    The rates of growth and regeneration were compared for compact callus, friable callus, and suspension cells of Lilium longiflorum cv. Nellie White to determine the optimal culture conditions. The fresh weight was higher for compact callus induced from bulb scales cultured on Murashige and Skoog’s m...

  7. Effect of sorbitol in callus induction and plant regeneration in wheat ...

    African Journals Online (AJOL)

    Six wheat genotypes were evaluated for their response to callus induction and regeneration on MS medium modified with different concentrations of sorbitol, that is, 0, 10, 20, 30 gL-1 along with optimum (3 mgL-1) concentration of 2,4-D. Variability was observed among different genotypes for callus induction. Highest callus ...

  8. Which stem parts of Slender speedwell (Veronica filiformis) are the most successful in plant regeneration?

    Czech Academy of Sciences Publication Activity Database

    Šerá, Božena

    2012-01-01

    Roč. 67, č. 1 (2012), s. 110-115 ISSN 0006-3088 R&D Projects: GA MŠk OC10032 Institutional research plan: CEZ:AV0Z60870520 Keywords : clonal * invasive species * regeneration * shoot * stem * terminal * vegetative reproduciton Subject RIV: EF - Botanics Impact factor: 0.506, year: 2012

  9. Effect of NaCl on in vitro plant regeneration from embryogenic callus ...

    African Journals Online (AJOL)

    In vitro experiments were conducted to assess the effect of salt stress on callus induction, survival, fresh weight, regeneration, proline level and total protein ... that proline accumulation is an index of salinity tolerance and this important international variety can be genetically manipulated to develop salinity tolerant crop.

  10. Rapid acceleration of plant speciation during the Anthropocene.

    Science.gov (United States)

    Thomas, Chris D

    2015-08-01

    Speciation rates need to be considered when estimating human impacts on the numbers of species on Earth, given that past mass extinctions have been followed by the accelerated origination of new taxa. Here, I suggest that the Anthropocene is already exhibiting a greatly accelerated plant speciation rate due to agriculture, horticulture, and the human-mediated transport of species, followed by hybridisation. For example, more new plant species have come into existence in Europe over the past three centuries than have been documented as becoming extinct over the same period, even though most new hybrid-origin species are likely to remain undetected. Current speciation rates are unusually high and they could be higher than during or after previous mass extinctions. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Mobile zinc increases rapidly in the retina after optic nerve injury and regulates ganglion cell survival and optic nerve regeneration.

    Science.gov (United States)

    Li, Yiqing; Andereggen, Lukas; Yuki, Kenya; Omura, Kumiko; Yin, Yuqin; Gilbert, Hui-Ya; Erdogan, Burcu; Asdourian, Maria S; Shrock, Christine; de Lima, Silmara; Apfel, Ulf-Peter; Zhuo, Yehong; Hershfinkel, Michal; Lippard, Stephen J; Rosenberg, Paul A; Benowitz, Larry

    2017-01-10

    Retinal ganglion cells (RGCs), the projection neurons of the eye, cannot regenerate their axons once the optic nerve has been injured and soon begin to die. Whereas RGC death and regenerative failure are widely viewed as being cell-autonomous or influenced by various types of glia, we report here that the dysregulation of mobile zinc (Zn(2+)) in retinal interneurons is a primary factor. Within an hour after the optic nerve is injured, Zn(2+) increases several-fold in retinal amacrine cell processes and continues to rise over the first day, then transfers slowly to RGCs via vesicular release. Zn(2+) accumulation in amacrine cell processes involves the Zn(2+) transporter protein ZnT-3, and deletion of slc30a3, the gene encoding ZnT-3, promotes RGC survival and axon regeneration. Intravitreal injection of Zn(2+) chelators enables many RGCs to survive for months after nerve injury and regenerate axons, and enhances the prosurvival and regenerative effects of deleting the gene for phosphatase and tensin homolog (pten). Importantly, the therapeutic window for Zn(2+) chelation extends for several days after nerve injury. These results show that retinal Zn(2+) dysregulation is a major factor limiting the survival and regenerative capacity of injured RGCs, and point to Zn(2+) chelation as a strategy to promote long-term RGC protection and enhance axon regeneration.

  12. Collision-induced fragmentation accurate mass spectrometric analysis methods to rapidly characterize plant extracts

    Science.gov (United States)

    The rapid advances in analytical chromatography equipment have made the reliable and reproducible measurement of a wide range of plant chemical components possible. Full chemical characterization of a given plant material is possible with the new mass spectrometers currently available. For phytochem...

  13. Collision-induced fragmentation accurate mass spectrometric analysis methods to rapidly characterize phytochemicals in plant extracts

    Science.gov (United States)

    The rapid advances in analytical chromatography equipment have made the reliable and reproducible measurement of a wide range of plant chemical components possible. Full chemical characterization of a given plant material is possible with the new mass spectrometers currently available. New methods a...

  14. In vitro regeneration in Sarcostemma acidum (Roxb.) -an important medicinal plant of semi-arid ecosystem of Rajasthan, India.

    Science.gov (United States)

    Rathore, Mahender S; Shekhawat, Narpat S

    2013-04-01

    An efficient regeneration protocol for Sarcostemma acidum - an important medicinal plant has been established. Callus initiated from nodal explant on MS medium with 2.0 mg L(-1) of NAA + additives. Callus initiated was subcultured on MS medium containing various concentrations of NAA or 2,4-D. Out of these combinations, MS medium +1.0 mg L(-1) of NAA + additives was found to be effective for the multiplication of callus. Subculture was done after an interval of 20-22 days. For differentiation of callus BAP or Kinetin alone was found to be less effective. Maximum frequency of shoot regeneration recorded on MS medium +1.0 mg L(-1) of BAP + 0.5 mg L(-1) of Kinetin and 0.1 mg L(-1) of NAA + additives. The in vitro differentiated shoots were excised and inoculated on 1/4 strength MS medium +2.0 mg L(-1) of IBA + 0.02 % activated charcoal for in vitro rooting. Maximum response (90 %) was recorded on this medium. In vitro differentiated shoots were inoculated on autoclaved soilrite® after treatment with root inducing auxins. Ex vitro rooting in this plant species has been reported for the first time. Eighty five percent of the shoots rooted under ex vitro conditions. Both in vitro and ex vitro rooted plantlets were hardened in a green house.

  15. Improved plant regeneration from cultured leaf segments in peanut (Arachis hypogaea L.) by limited exposure to thidiazuron.

    Science.gov (United States)

    Akasaka; Daimon; Mii

    2000-07-28

    Bud primordia were induced from leaf segments, which were harvested from young seedlings of Spanish type peanut (Arachis hypogaea L. cv. Chico), on 0.8% agar-solidified medium containing Murashige and Skoog (MS) basal salts supplemented with B5 vitamins, 1 mg/l NAA and various cytokinins such as benzyladenine (BA), isopentenyladenine (2ip), kinetin (KIN), chloropyridylphenylurea (4PU), thidiazuron (TDZ), zeatin (ZTN) in different concentrations. Among the cytokinins tested, TDZ was found to be the most efficient for inducing bud primordia. However, continuous culture on TDZ-containing media induced abnormal development of these primordia, and they failed to grow into plantlets. Histological observations revealed that the malformation most often obtained was a shoot-like structure which lacked shoot apical meristem (SAM) and had disorganized vascular bundles. For normal shoot regeneration, it was necessary to limit the culture period of the explants on TDZ-containing medium to 7 days at 10 mg/l or 21 days at 1 mg/l and then transfer them onto plant growth regulator-free medium. The percentage of conversion from shoot buds to shoots was 34.7%. When shoots were removed from the explants and transferred onto basal medium containing 1 mg/l NAA, all regenerated shoots readily rooted and successfully acclimatized. All of the acclimatized plants produced viable seeds in the greenhouse condition.

  16. Field evaluation of regenerated plants by somatic embryogenesis from shoots apexes of axillary buds in ´Navolean’ (Musa spp., AAB.

    Directory of Open Access Journals (Sweden)

    Jorge López

    2005-04-01

    Full Text Available The use of shoots apexes from axilary buds for callus induction with embryogenic structures in plantain ‘Navolean’ (Group AAB permitted to develop a plant regeneration method through out somatic embryogenesis. In order to know the phenotypic variants that may be produced with the previously mentioned method , 1000 plants were planted in field conditions in comparison to those coming from somatic embryos obtained from multibuds as initial explants and organogenesis-derived plants (shoot tipsand conventionally derived plants (corms, during two growing cycles. The main morphological characters and yield components were evaluated. The total frequency of somaclonal variation during the first growing cycle in plants coming from somatic embryos obtained from shoots apexes from axilary buds as initial explants were 1.1%, and 8,6% in regenerated plants from somatic embryos obtained from multi-buds as initial explants. Later, in this same growing cycle, plants regenerated from somatic embryos (both sources showed a similar performance between them and they were significantly superior in all evaluated variants in comparison to corm-derived plants. In the second growing cycle, significant differences were not observed in yield components of suckers from evaluated plants, in spite of the propagation method used. With regard to somaclonal variation, the best performance was obtained with shoots apexes from axilary buds as explants. Finally, the feasibility of using the new method was shown. Key words: embryogenic cell suspensions, somaclonal variation

  17. Biomécanique de mouvements rapides chez les plantes

    OpenAIRE

    Llorens, Coraline

    2014-01-01

    In this PhD work, we focus on the biomechanics of two motions among the fastest in plant kingdom. The first part is a theoretical study of the motion leading to a prey capture by the bladderwort’s traps, elastic millimeter-sized bladders closed by a flexible door. A dynamical model, based on mechanical, elastic and hydrodynamic ingredients, links the pressure difference between the trap and its surroundings with the door position by the means of two coupled ordinary differential equations. Th...

  18. NLR-parser: rapid annotation of plant NLR complements.

    Science.gov (United States)

    Steuernagel, Burkhard; Jupe, Florian; Witek, Kamil; Jones, Jonathan D G; Wulff, Brande B H

    2015-05-15

    The repetitive nature of plant disease resistance genes encoding for nucleotide-binding leucine-rich repeat (NLR) proteins hampers their prediction with standard gene annotation software. Motif alignment and search tool (MAST) has previously been reported as a tool to support annotation of NLR-encoding genes. However, the decision if a motif combination represents an NLR protein was entirely manual. The NLR-parser pipeline is designed to use the MAST output from six-frame translated amino acid sequences and filters for predefined biologically curated motif compositions. Input reads can be derived from, for example, raw long-read sequencing data or contigs and scaffolds coming from plant genome projects. The output is a tab-separated file with information on start and frame of the first NLR specific motif, whether the identified sequence is a TNL or CNL, potentially full or fragmented. In addition, the output of the NB-ARC domain sequence can directly be used for phylogenetic analyses. In comparison to other prediction software, the highly complex NB-ARC domain is described in detail using several individual motifs. © The Author 2015. Published by Oxford University Press.

  19. Conceptual design of a coal-fired MHD retrofit plant. Topical report, Seed Regeneration System Study 2

    Energy Technology Data Exchange (ETDEWEB)

    1992-11-01

    Westinghouse Advanced Energy Systems (WAES), through Contract No. DE-AC22-87PC79668 funded by US DOE/PETC, is conducting a conceptual design study to evaluate a coal-fired magnetohydrodynamic (MHD) retrofit of a utility plant of sufficient size to demonstrate the technical and future economic viability of an MHD system operating within an electric utility environment. The objective of this topical report is to document continuing seed regeneration system application studies and the definition of will system integration requirements for the Scholz MHD retrofit plant design. MHD power plants require the addition of a seeding material in the form of potassium to enhance the ionization of the high temperature combustion gas in the MHD channel. This process has an added environmental advantage compared to other types of coal-fired power plants in that the potassium combines with the naturally occurring sulfur in the coal to form a potassium sulfate flyash (K{sub 2}SO{sub 4}) which can be removed from the process by appropriate particulate control equipment. Up to 100% of the Sulfur in the coal can be removed by this process thereby providing environmentally clean power plant operation that is better than required by present and anticipated future New Source Performance Standards (NSPS).

  20. Ability of Callus Induction and Plant Regeneration in White Kashan and Red Rey Onions Cultivars (Allium cepa L. Using Root - tip Culture under Invitro Conditions

    Directory of Open Access Journals (Sweden)

    A. Goravanchi

    2011-01-01

    Full Text Available This present study has been carried out in order to evaluate the effect of culture medium and genotype on callus induction and regeneration of two onion varieties through root tip culture. For callus induction, micro samples from 1-3 mm root tip of 2 day old in vitro germinated plants laied on four culture media (1mg/l 2, 4-D, 0/5mg/l 2, 4-D, 0/5mg/l 2, 4-D with 0/5mg/l kinetin, 1mg/l 2,4-D with 1mg/l kinetin in darkness conditions. Subculture of samples was done after four weeks. The  produced calluses were laied in embryogenic culture medium for 4 weeks and after formation of embryoes, they were put in plant regeneration culture medium for eight weeks. Result showed that callus induction percentage is affected significantly by cultivar and culture medium, but reciprocal effect of both factors was not meaningful on callus induction percentage. Plant regeneration percentage and number of embryo per callus is affected meaningfully by cultivar, but type of culture medium and reciprocal effect of both factors was not meaningful on plant regeneration percentage and number of embryoes per callus. Embryogenic callus percentage is affected meaningfully by cultivar, but type of culture medium had not meaningful effect on embryogenic callus percentage. Callus induction percentage, plant regeneration percentage, number of embryo per callus and embryogenic callus percentage, are higher in white Kashan cultivar in relation to red Rey one and in culture medium involving hormonal combination of 2,4-D and kinetin, application of 0/5mg/l 2, 4-D can be usful. Considering the callus induction percentage, embryogenic and plant regeneration percentage, white Kashan cultivar and  culture medium containing 0/5mg/l 2, 4-D is appropriate as compared with other cultures.

  1. Immature tassels as alternative explants in somatic embryogenesis and plant regeneration in south Brazilian maize genotypes - doi: 10.4025/actasciagron.v35i1.15545

    Directory of Open Access Journals (Sweden)

    Magali Ferrari Grando

    2012-08-01

    Full Text Available Somatic embryogenesis and in vitro plant regeneration are fundamental processes in the obtainment of transgenic maize plants. Explant, genotype and culture medium are determining factors in these processes. Immature embryo explants and the American Hi-II genotype have been widely employed to acquire genetically modified plants in this species. However, the use of more readily available explants is desired as well as the development of genetic transformation protocols for productive genotypes adapted to local conditions. This study provides an evaluation of immature tassel explants in relation to embryogenic callus production and plant regeneration in South Brazilian maize genotypes for their use in genetic transformation experiments. Immature tassels from 5 hybrids were cultivated in different callus-induction media. The frequency and the fresh mass of embryogenic calli were evaluated. The frequency was influenced by genotype, and the fresh mass was influenced by genotype and culture medium. In plant regeneration, shoots, complete seedlings and acclimatized and fertile plants were quantified. Treatments producing long term embryogenic calli from immature tassels of South Brazilian genotypes with the capacity to regenerate were identified.

  2. Role of TDZ in the quick regeneration of multiple shoots from nodal explant of Vitex trifolia L.--an important medicinal plant.

    Science.gov (United States)

    Ahmed, Md Rafique; Anis, Mohammad

    2012-11-01

    The effect of thidiazuron (TDZ) has been investigated in shoot multiplication for a simple, efficient, rapid, and commercially applicable regeneration protocol of an important medicinal plant, Vitex trifolia. Multiple shoots were induced in nodal explants obtained from a mature tree on Murashige and Skoog (MS) medium supplemented with TDZ in various concentrations (0.5, 1.0, 2.5, 5.0, 7.5, or 10.0 μM). Prolonged exposure of the culture to TDZ had an adverse affect. To avoid this, the cultures were transferred to TDZ-free MS medium or MS medium fortified with various concentrations of 6-benzyladenine (BA) alone or in combination with α-naphthalene acetic acid (NAA) to enhance multiplication, proliferation, and elongation of induced shoots. Optimum shoot multiplication and elongation was achieved when TDZ-exposed explants were repeatedly subcultured on MS media containing a combination of 1.0 μM BA and 0.5 μM NAA. The highest shoot regeneration frequency (90 %) and maximum number (22.3 ± 0.2) of shoots per explant with shoot length of (5.2 ± 0.2 cm) was recorded on MS medium fortified with 5.0 μM TDZ. In vitro rooting of isolated shoots was achieved best in half-strength MS medium containing 0.5 μM NAA. Properly rooted plantlets were successfully hardened off and acclimatized in thermocol cups containing sterile Soilrite. These plantlets were then transferred to pots containing different potting substrate; percentage survival of the plantlets was highest in vermiculite/garden soil mixture (1:1) and successfully transfer to greenhouse under sunlight.

  3. An efficient in vitro plantlet regeneration from shoot tip cultures of Curculigo latifolia, a medicinal plant.

    Science.gov (United States)

    Babaei, Nahid; Abdullah, Nur Ashikin Psyquay; Saleh, Ghizan; Abdullah, Thohirah Lee

    2014-01-01

    A procedure was developed for in vitro propagation of Curculigo latifolia through shoot tip culture. Direct regeneration and indirect scalp induction of Curculigo latifolia were obtained from shoot tip grown on MS medium supplemented with different concentrations and combinations of thidiazuron and indole-3-butyric acid. Maximum response for direct regeneration in terms of percentage of explants producing shoot, shoot number, and shoot length was obtained on MS medium supplemented with combination of thidiazuron (0.5 mg L(-1)) and indole-3-butyric acid (0.25 mg L(-1)) after both 10 and 14 weeks of cultures. Indole-3-butyric acid in combination with thidiazuron exhibited a synergistic effect on shoot regeneration. The shoot tips were able to induce maximum scalp from basal end of explants on the medium with 2 mg L(-1) thidiazuron. Cultures showed that shoot number, shoot length, and scalp size increased significantly after 14 weeks of culture. Transferring of the shoots onto the MS medium devoid of growth regulators resulted in the highest percentage of root induction and longer roots, while medium supplemented with 0.25 mg L(-1) IBA produced more numbers of roots.

  4. Plant Thioredoxin CDSP32 Regenerates 1-Cys Methionine Sulfoxide Reductase B Activity through the Direct Reduction of Sulfenic Acid*

    Science.gov (United States)

    Tarrago, Lionel; Laugier, Edith; Zaffagnini, Mirko; Marchand, Christophe H.; Le Maréchal, Pierre; Lemaire, Stéphane D.; Rey, Pascal

    2010-01-01

    Thioredoxins (Trxs) are ubiquitous enzymes catalyzing the reduction of disulfide bonds, thanks to a CXXC active site. Among their substrates, 2-Cys methionine sulfoxide reductases B (2-Cys MSRBs) reduce the R diastereoisomer of methionine sulfoxide (MetSO) and possess two redox-active Cys as follows: a catalytic Cys reducing MetSO and a resolving one, involved in disulfide bridge formation. The other MSRB type, 1-Cys MSRBs, possesses only the catalytic Cys, and their regeneration mechanisms by Trxs remain unclear. The plant plastidial Trx CDSP32 is able to provide 1-Cys MSRB with electrons. CDSP32 includes two Trx modules with one potential active site 219CGPC222 and three extra Cys. Here, we investigated the redox properties of recombinant Arabidopsis CDSP32 and delineated the biochemical mechanisms of MSRB regeneration by CDSP32. Free thiol titration and 4-acetamido-4′-maleimidyldistilbene-2,2′-disulfonic acid alkylation assays indicated that the Trx possesses only two redox-active Cys, very likely the Cys219 and Cys222. Protein electrophoresis analyses coupled to mass spectrometry revealed that CDSP32 forms a heterodimeric complex with MSRB1 via reduction of the sulfenic acid formed on MSRB1 catalytic Cys after MetSO reduction. MSR activity assays using variable CDSP32 amounts revealed that MSRB1 reduction proceeds with a 1:1 stoichiometry, and redox titrations indicated that CDSP32 and MSRB1 possess midpoints potentials of −337 and −328 mV at pH 7.9, respectively, indicating that regeneration of MSRB1 activity by the Trx through sulfenic acid reduction is thermodynamically feasible in physiological conditions. PMID:20236937

  5. High frequency plant regeneration from mature seed of elite, recalcitrant Malaysian indica rice ( Oryza sativa L.) CV. MR 219.

    Science.gov (United States)

    Sivakumar, P; Law, Y S; Ho, C-L; Harikrishna, Jennifer Ann

    2010-09-01

    An efficient in vitro plant regeneration system was established for elite, recalcitrant Malaysian indica rice, Oryza sativa L. CV. MR 219 using mature seeds as explant on Murashige and Skoog and Chu N6 media containing 2,4-dichlorophenoxy acetic acid and kinetin either alone or in different combinations. L-proline, casein hydrolysate and L-glutamine were added to callus induction media for enhancement of embryogenic callus induction. The highest frequency of friable callus induction (84%) was observed in N6 medium containing 2.5 mg l(-1) 2,4-dichlorophenoxy acetic acid, 0.2 mg l(-1) kinetin, 2.5 mg l(-1) L-proline, 300 mg l(-1) casein hydrolysate, 20 mg l(-1) L-glutamine and 30 g l(-1) sucrose under culture in continuous lighting conditions. The maximum regeneration frequency (71%) was observed, when 30-day-old N6 friable calli were cultured on MS medium supplemented with 3 mg l(-1) 6-benzyl aminopurine, 1 mg l(-1) naphthalene acetic acid, 2.5 mg l(-1) L-proline, 300 mg l(-1) casein hydrolysate and 3% maltose. Developed shoots were rooted in half strength MS medium supplemented with 2% sucrose and were successfully transplanted to soil with 95% survival. This protocol may be used for other recalcitrant indica rice genotypes and to transfer desirable genes in to Malaysian indica rice cultivar MR219 for crop improvement.

  6. Analysis of somaclonal variation in transgenic and regenerated plants of Arabidopsis thaliana using methylation related metAFLP and TMD markers.

    Science.gov (United States)

    Coronel, Carlos J; González, Ana I; Ruiz, María L; Polanco, Carlos

    2018-01-01

    We provide evidence that nucleotide sequence and methylation status changes occur in the Arabidopsis genome during in vitro tissue culture at a frequency high enough to represent an important source of variation. Somaclonal variation is a general consequence of the tissue culture process that has to be analyzed specifically when regenerated plants are obtained in any plant species. Currently, there are few studies about the variability comprising sequence changes and methylation status at the DNA level, generated by the culture of A. thaliana cells and tissues. In this work, two types of highly reproducible molecular markers, modified methylation sensitive AFLP (metAFLP) and transposon methylation display (TMD) have been used for the first time in this species to analyze the nucleotide and cytosine methylation changes induced by transformation and tissue culture protocols. We found significantly higher average methylation values (7.5%) in regenerated and transgenic plants when compared to values obtained from seed derived plants (3.2%) and that the main component of the somaclonal variation present in Arabidopsis clonal plants is genetic rather than epigenetic. However, we have found that the Arabidopsis regenerated and transgenic plants had a higher number of non-fully methylated sites flanking transposable elements than the control plants, and therefore, their mobilization can be facilitated. These data provide further evidence that changes in nucleotide sequence and methylation status occur in the Arabidopsis genome during in vitro tissue culture frequently enough to be an important source of variation in this species.

  7. Evaluation of antimicrobial activity of selected plant extracts by rapid XTT colorimetry and bacterial enumeration.

    Science.gov (United States)

    Al-Bakri, Amal G; Afifi, Fatma U

    2007-01-01

    The aim of this study was to screen and evaluate the antimicrobial activity of indigenous Jordanian plant extracts, dissolved in dimethylsulfoxide, using the rapid XTT assay and viable count methods. XTT rapid assay was used for the initial screening of antimicrobial activity for the plant extracts. Antimicrobial activity of potentially active plant extracts was further assessed using the "viable plate count" method. Four degrees of antimicrobial activity (high, moderate, weak and inactive) against Bacillus subtilis, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa, respectively, were recorded. The plant extracts of Hypericum triquetrifolium, Ballota undulata, Ruta chalepensis, Ononis natrix, Paronychia argentea and Marrubium vulgare had shown promising antimicrobial activity. This study showed that while both XTT and viable count methods are comparable when estimating the overall antimicrobial activity of experimental substances, there is no strong linear correlation between the two methods.

  8. Carbon source dependent somatic embryogenesis and plant regeneration in cotton, Gossypium hirsutum L. cv. SVPR2 through suspension cultures.

    Science.gov (United States)

    Ganesan, M; Jayabalan, N

    2005-10-01

    Highly reproducible and simple protocol for cotton somatic embryogenesis is described here by using different concentrations of maltose, glucose, sucrose and fructose. Maltose (30 g/l) is the best carbon source for embryogenic callus induction and glucose (30 g/l) was suitable for induction, maturation of embryoids and plant regeneration. Creamy white embryogenic calli of hypocotyl explants were formed on medium containing MS basal salts, myo-inositol (100 mg/l), thiamine HCI (0.3 mg/l), picloram (0.3 mg/l), Kin (0.1 mg/l) and maltose (30 g/l). During embryo induction and maturation, accelerated growth was observed in liquid medium containing NH3NO4 (1 g/l), picloram (2.0 mg/l), 2 ip (0.2 mg/l), Kin (0.1 mg/l) and glucose (30 g/l). Before embryoid induction, large clumps of embryogenic tissue were formed. These tissues only produced viable embryoids. Completely matured somatic embryos were germinated successfully on the medium fortified with MS salts, myo-inositol (50 mg/l), thiamine HCl (0.2 mg/l), GA3 (0.2 mg/l), BA (1.0 mg/l) and glucose (30 g/l). Compared with earlier reports, 65% of somatic embryo germination was observed. The abnormal embryo formation was highly reduced by using glucose (30 g/l) compared to other carbon sources. The regenerated plantlets were fertile but smaller in height than the seed derived control plants.

  9. The development of an efficient cultivar-independent plant regeneration system from callus derived from both apical and non-apical root segments of garlic (Allium sativum L.)

    NARCIS (Netherlands)

    Zheng, S.J.; Henken, G.; Krens, F.A.; Kik, C.

    2003-01-01

    Callus induction and later plant regeneration were studied in four widely grown garlic (Allium sativum L.) cultivars from Europe. Root segments from in vitro plantlets were used as starting material. In addition to cultivar effects, the effects of auxin and cytokinin levels and the position of the

  10. Agrobacterium tumefaciens-mediated transgenic plant and somaclone production through direct and indirect regeneration from leaves in Stevia rebaudiana with their glycoside profile.

    Science.gov (United States)

    Khan, Shamshad Ahmad; Ur Rahman, Laiq; Shanker, Karuna; Singh, Manju

    2014-05-01

    Agrobacterium tumefaciens (EHA-105 harboring pCAMBIA 1304)-mediated transgenic plant production via direct regeneration from leaf and elite somaclones generation through indirect regeneration in Stevia rebaudiana is reported. Optimum direct regeneration frequency along with highest transformation frequency was found on MS + 1 mg/l BAP + 1 mg/l NAA, while indirect regeneration from callus was obtained on MS + 1 mg/l BAP + 2 mg/l NAA. Successful transfer of GUS-positive (GUS assay and PCR-based confirmation) transgenic as well as four somaclones up to glasshouse acclimatization has been achieved. Inter-simple sequence repeat (ISSR) profiling of transgenic and somaclonal plants showed a total of 113 bands, out of which 49 were monomorphic (43.36 %) and 64 were polymorphic (56.64 %). Transgenic plant was found to be closer to mother plant, while on the basis of steviol, stevioside, and rebaudioside A profile, somaclone S2 was found to be the best and showed maximum variability in ISSR profiling.

  11. Efficient callus induction and plant regeneration from anther of Chinese narcissus (Narcissus tazetta L. var. chinensis Roem).

    Science.gov (United States)

    Chen, LinJiao; Zhu, XueYi; Gu, Li; Wu, Jian

    2005-09-01

    Callus culture has, to date, been reported only in a few species of Narcissus. We used anthers of Chinese narcissus (Narcissus tazetta L. var. chinensis Roem) as explants for callus induction and plant regeneration. A high percentage of anthers at the early- to mid-uninucleate microspore stage were responsive on the basal MS medium supplemented with 0.5-1 mg l(-1) 2,4-dichlorophenoxyacetic acid and 0.5-2 mg l(-1) 6-benzyladenine under dark conditions. Calli were initiated from anther connective tissue or anther wall tissue, and no division of microspores occurred during callus formation, as determined by histological observation. Using 20 random amplified polymorphic DNA primers, we verified the genetic integrity of the anther-derived plants of Chinese narcissus with respect to the donor plants. These results suggest that anther culture in vitro can provide an efficient new micropropagation technique for Chinese narcissus as well as a new strategy for in vitro mass propagation of other daffodils.

  12. Rapid and reliable extraction of genomic DNA from various wild-type and transgenic plants

    Directory of Open Access Journals (Sweden)

    Yang Moon-Sik

    2004-09-01

    Full Text Available Abstract Background DNA extraction methods for PCR-quality DNA from calluses and plants are not time efficient, since they require that the tissues be ground in liquid nitrogen, followed by precipitation of the DNA pellet in ethanol, washing and drying the pellet, etc. The need for a rapid and simple procedure is urgent, especially when hundreds of samples need to be analyzed. Here, we describe a simple and efficient method of isolating high-quality genomic DNA for PCR amplification and enzyme digestion from calluses, various wild-type and transgenic plants. Results We developed new rapid and reliable genomic DNA extraction method. With our developed method, plant genomic DNA extraction could be performed within 30 min. The method was as follows. Plant tissue was homogenized with salt DNA extraction buffer using hand-operated homogenizer and extracted by phenol:chloroform:isoamyl alcohol (25:24:1. After centrifugation, the supernatant was directly used for DNA template for PCR, resulting in successful amplification for RAPD from various sources of plants and specific foreign genes from transgenic plants. After precipitating the supernatant, the DNA was completely digested by restriction enzymes. Conclusion This DNA extraction procedure promises simplicity, speed, and efficiency, both in terms of time and the amount of plant sample required. In addition, this method does not require expensive facilities for plant genomic DNA extraction.

  13. Indirect regeneration from in vitro leaf tissue of periwinkle (Catharanthus roseus L.) in response to different treatments of plant growth regulators

    OpenAIRE

    B.E. Sayed-Tabatabaei; F. Eatesam; M. Talebi

    2012-01-01

    Periwinkle (Catharanthus roseus L.) belongs to the Apocynaceae family and accumulates more than 130 terpenoid indole alkaloids (TIAs), of which two dimeric alkaloids Vinblastine and Vincristine have antineoplastic activity and are useful for treatment of various cancers. Therefore, the production of these drugs has been emphasized in plant tissue culture. In this research, 25 treatments of plant growth regulators to produce callus from leaf explants and seven treatments for regeneration of ca...

  14. Callus induction and plant regeneration from different explant types of Miscanthus x ogiformis Honda 'Giganteus'

    DEFF Research Database (Denmark)

    Holme, Inger Bæksted; Petersen, Karen Koefoed

    1996-01-01

    Different explants of Miscanthus x ogiformis Honda 'Giganteus' were tested in order to develop an efficient tissue culture system. Shoot apices, leaf and root sections from in vitro-propagated plants, and leaf and immature inflorescence sections from 6-month-old greenhouse-grown plants were used....

  15. Factors influencing plant regeneration from seedling explants of Hairy nightshade (Solanum sarrachoides)

    Science.gov (United States)

    A good model plant to investigate plant – pathogen interactions would be easy to grow, have a short life cycle, be a natural host of many pathogens, and be easy to manipulate genetically. Hairy nightshade (Solanum sarrachoides) is a ubiquitous, fast growing weed that produces copious amounts of see...

  16. Evaluating different planting stocks for oak regeneration on Hurricane Katrina disturbed lands

    Science.gov (United States)

    Damon B. Hollis; Andrew W. Ezell; Emily B. Schultz; John D. Hodges; Andrew B. Self; Derek K. Alkire

    2012-01-01

    Three oak planting stocks were evaluated to determine their influence on survival and initial growth. Planting stocks utilized included conventional containerized seedlings in 240 centimeter3 (cm3) containers, 1-0, bareroot seedlings, and Root Production Method (RPM™) seedlings in 11.4 liter (L) containers. Initial height, groundline diameter (...

  17. Plant regeneration from protoplasts of Panax ginseng (C.A. Meyer) through somatic embryogenesis.

    Science.gov (United States)

    Arya, S; Liu, J R; Eriksson, T

    1991-09-01

    Protoplasts of Panax ginseng were isolated from embryos obtained from the 4-year old embryogenic cell line KCTC PCL 49031 which was derived from a zygotic embryo. High protoplast yields of 22-25 × 10(6) protoplast / g tissue were obtained following 5-6 h digestion with 2% Cellulysin, 1% Pectinase and 1% Macerasae in half strength Murashige and Skoog's medium containing 12% mannitol. A plating density of 1×10(5) protoplasts /ml was found optimal for protoplast culture. An initial division frequency of 10% was obtained in an agarosegelled defined medium. Myo-inositol (6%) was found to be the most suitable osmoticum. Somatic embryos were formed from protoplast derived embryogenic callus, which regenerated into plantlets.

  18. Evaluation of peanut genotypes for in vitro plant regeneration using thidiazuron.

    Science.gov (United States)

    Matand, Kanyand; Prakash, C S

    2007-06-15

    A major limitation with the available protocols for in vitro regeneration of peanut (Arachis hypogaea L.) is their narrow application to very few select genotypes. Here, we report a protocol that can be applied across a broad spectrum of peanut market types, explant types and geographic regions using thidiazuron (TDZ). The effect of the timing of TDZ application to the culturing of both zygotic embryos and subsequent plantlet explants on MS medium is also reported. An extended use of TDZ and at a higher concentration (30 m/l) resulted in the greatest explant shoot average (approximately 13). However, a limited application of TDZ (10 d) was sufficient to induce shoot formation in peanut. Hypocotyl was the best explant type that induced the greatest shoot average (15) across market types followed by lamina (7.4). Spanish and Valencia were the most efficient market groups that induced shoots across explant types, consistently.

  19. Effects of N6-benzylaminopurine and Indole Acetic Acid on In Vitro Shoot Multiplication, Nodule-like Meristem Proliferation and Plant Regeneration of Malaysian Bananas (Musa spp.)

    Science.gov (United States)

    Sipen, Philip; Davey, Michael R

    2012-01-01

    Different concentrations of N6-benzylaminopurine (BAP) and indole acetic acid (IAA) in Murashige and Skoog based medium were assessed for their effects on shoot multiplication, nodule-like meristem proliferation and plant regeneration of the Malaysian banana cultivars Pisang Mas, Pisang Nangka, Pisang Berangan and Pisang Awak. BAP at 1–14 mg L−1 with or without 0.2 mg L−1 IAA, or BAP at 7–14 mg L−1 with the same concentration of IAA, was evaluated for shoot multiplication from shoot tips and the proliferation of nodule-like meristems from scalps, respectively. Plant regeneration from scalps was assessed using 1 mg L−1 BAP and 0.2 mg L−1 IAA separately, or a combination of these two growth regulators. Data on shoot multiplication, the proliferation of nodule-like meristems with associated plant regeneration were recorded after 30 days of culture. A maximum of 5 shoots per original shoot tip was achieved on medium supplemented with BAP at 5 mg L−1 (Pisang Nangka), 6 mg L−1 (Pisang Mas and Pisang Berangan), or 7 mg L−1 (Pisang Awak), with 0.2 mg L−1 IAA. BAP at 11 mg L−1 with 0.2 mg L−1 IAA induced the most highly proliferating nodule-like meristems in the four banana cultivars. Plant regeneration from scalps was optimum in all cases on medium containing 1 mg L−1 BAP and 0.2 mg L−1 IAA. This is the first report on the successful induction of highly proliferating nodule-like meristems and plant regeneration from scalps of the Malaysian banana cultivars Pisang Mas, Pisang Nangka, Pisang Berangan and Pisang Awak. PMID:24575235

  20. Assessment of Methods for Rapid Intraoperative Concentration and Selection of Marrow-Derived Connective Tissue Progenitors for Bone Regeneration Using the Canine Femoral Multidefect Model

    Science.gov (United States)

    Luangphakdy, Viviane; Boehm, Cynthia; Pan, Hui; Herrick, James; Zaveri, Phil

    2016-01-01

    Treatment of large bone defects remains an unsolved clinical challenge, despite a wide array of existing bone graft materials and strategies. Local deficiency in osteogenic connective tissue progenitors (CTP-Os) due to tissue loss is one of the central biological barriers to bone regeneration. Density separation (DS) and selective retention (SR) represent two promising methods that can be used intraoperatively to rapidly concentrate cells and potentially select CTP-Os. This project was designed to compare DS and SR using the canine femoral multidefect (CFMD) model. Mineralized cancellous allograft (MCA) was used as a standardized scaffold for cell transplantation. Two experiments were performed using a cohort of six animals in each comparison. In Cohort I, unprocessed bone marrow aspirate (BMA) clot was compared to DS processing. MCA combined with raw BMA or DS processed cells produced a robust and advanced stage of bone regeneration throughout the defect in 4 weeks with reconstitution of hematopoietic marrow. However, the retention of DS processed cells and CTP-Os in the MCA matrix was low compared to BMA clot. In Cohort II, MCA with DS-T cells (addition of calcium chloride thrombin to induce clotting and enhance cell and CTP-O retention) was compared to MCA with SR cells. A mean of 276 ± 86 million nucleated cells and 29,030 ± 10,510 CTP-Os were implanted per defect in the DS-T group. A mean of 76 ± 42 million nucleated cells and 30,266 ± 15,850 CTP-Os were implanted in the SR group. Bone formation was robust and not different between treatments. Histologically, both groups demonstrated regeneration of hematopoietic marrow tissue. However, SR sites contained more hematopoietic vascular tissues, less fibrosis, and less residual allograft, particularly in the intramedullary cavity, suggesting a more advanced stage of remodeling (p = 0.04). These data demonstrate excellent overall performance of DS and SR processing methods. Both methods

  1. Plant regeneration and Agrobacterium-mediated transformation of Vacuolar H+-ATPase c Subunit Gene in hybrid poplar populus davidiana Dode × P. bollena Lauche

    Directory of Open Access Journals (Sweden)

    Han Xue

    2017-01-01

    Full Text Available An efficient regeneration and transformation system was developed for hybrid poplar populus davidiana Dode × P. bollena Lauche. Several factors, such as a competent regeneration protocol, antibiotics, bacterial concentration, infection time have been shown to affect transformation efficiency. It is the first report describing an efficient protocol for Vacuolar H+-ATPase c Subunit Gene from salt tolerant plant Puccinellia chinampoensis, which transformed into populus. Stable transgene integration was confirmed by the expression of GFP fusion proteins in leaves, stem and root for populus.

  2. In vitro regeneration and Agrobacterium tumefaciens-mediated genetic transformation in asakura-sanshoo (Zanthoxylum piperitum (L.) DC. F. inerme Makino) an important medicinal plant

    Science.gov (United States)

    Zeng, Xiaofang; Zhao, Degang

    2015-01-01

    Context: Asakura-sanshoo (Zanthoxylum piperitum [L.] DC. f. inerme Makino) is an important medicinal plant in East Asia. Transgenic technique could be applied to improve plant traits and analyze gene function. However, there is no report on regeneration and genetic transformation in Asakura-sanshoo. Aims: To establish a regeneration and Agrobacterium tumefaciens-mediated genetic transformation system in Asakura-sanshoo, which could be used for cultivar improvement and gene function analysis. Settings and Design: The various combinations of indole-3-butyric acid (IBA), 6-benzylaminopurine (BA) and naphthalene acetic acid (NAA) were explored for the optimal plant regeneration from petiole and stem of Asakura-sanshoo. The half-strength woody plant medium (WPM) with different concentrations of NAA and IBA was used to induce root. For genetic transformation, A. tumefaciens strain EHA-105 harboring the plasmid pBin-Ex-H-ipt which carries the isopentenyl transferase (ipt) gene, β-glucuronidase (GUS) gene and kanamycin resistance gene neomycin phosphotransferase II (NPTII) were used. The transformation efficiency was detected by the kanamycin resistant frequency. Materials and Methods: Petioles and stems were obtained from the in vitro cultured Asakura-sanshoo. The petiole and stem segments were precultured for 3 days, and then inflected using the bacterium at the concentration of OD600 0.5–0.8 for 10 min, followed by 3 days co-cultivation. Selection of the transgenic plants was carried out after 7 days the regeneration using gradient kanamycin at 30 mg/L and 50 mg/L, respectively. Successful transformed plants were confirmed by GUS histochemical assays, polymerase chain reaction (PCR), reverse transcription-PCR (RT-PCR), and Southern blotting analysis. Results: The highest shoots regeneration was obtained on WPM supplement with 0.5 mg/L BA and 0.2 mg/L NAA. The optimal rooting medium was half strength macro-element WPM. The kanamycin resistant frequency of petiole and

  3. Effects of Boreal Well Site Reclamation Practices on Long-Term Planted Spruce and Deciduous Tree Regeneration

    Directory of Open Access Journals (Sweden)

    Laurie A. Frerichs

    2017-06-01

    Full Text Available Well site development associated with oil sands exploration is common in boreal mixedwood forests of northern Alberta, Canada, and necessitates reforestation to accommodate other land uses. Little is known about the impact of soil and debris handling strategies during well site construction on long-term forest regeneration. This study addresses the impact of soil disturbance intensity, debris treatment, soil storage, and planting on the reforestation of 33 well sites reclaimed prior to 2006. Data on the survival and growth of planted white spruce (Picea glauca (Moench Voss and the regeneration density of deciduous trees, including trembling aspen (Populus tremuloides Michx, are presented from 2014 to 2015. The survival of planted spruce increased from 81% to 88% at well sites with a high relative to low soil disturbance. The total tree densities were lower in most treatments (≤2.69 stems m−2 than those in clear cuts (5.17 stems m−2, with the exception of root salvage areas where clear cuts had greater balsam poplar (Populus balsamifera L. densities (2.05 stems m−2 vs. <0.71 stems m−2 on all other treatments. Aspen densities were up to five times greater at well sites with low disturbance when compared to those with high disturbance, and this was further aided by shallow mulch at low disturbance sites. Spruce growth did not respond to well site treatments. Aspen growth (diameter and height remained similar between well site disturbance regimes; aspen exposed to high disturbance underperformed relative to low disturbance well sites and clear cut controls. With high disturbance, progressive soil piling led to increases in the density of aspen and birch (Betula papyrifera Marshall. Few long-term changes in soil were found due to well site development, with a greater soil pH in high disturbance sites compared to low disturbance sites. Overall, these results indicate that the nature of well site construction, including the extent of soil

  4. High-frequency shoot regeneration of nodal explants from ...

    African Journals Online (AJOL)

    Jane

    2011-06-29

    Jun 29, 2011 ... develop a rapid and efficient in vitro multiplication and regeneration system using nodal explants. MATERIALS AND METHODS. Plant material and initiation of in vitro shoot cultures. Young in vivo shoots with six to eight nodes of T. hemsleyanum were collected from wild population in Zhejiang Province, ...

  5. High-frequency plant regeneration through cyclic secondary somatic embryogenesis in black pepper (Piper nigrum L.).

    Science.gov (United States)

    Nair, R Ramakrishnan; Dutta Gupta, S

    2006-01-01

    A high-frequency plantlet regeneration protocol was developed for black pepper (Piper nigrum L.) through cyclic secondary somatic embryogenesis. Secondary embryos formed from the radicular end of the primary somatic embryos which were originally derived from micropylar tissues of germinating seeds on growth regulator-free SH medium in the absence of light. The process of secondary embryogenesis continued in a cyclic manner from the root pole of newly formed embryos resulting in clumps of somatic embryos. Strength of the medium and sucrose concentration influenced the process of secondary embryogenesis and fresh weight of somatic embryo clumps. Full-strength SH medium supplemented with 1.5% sucrose produced significantly higher fresh weight and numbers of secondary somatic embryos while 3.0 and 4.5% sucrose in the medium favored further development of proliferated embryos into plantlets. Ontogeny of secondary embryos was established by histological analysis. Secondary embryogenic potential was influenced by the developmental stage of the explanted somatic embryo and stages up to "torpedo" were more suitable. A single-flask system was standardized for proliferation, maturation, germination and conversion of secondary somatic embryos in suspension cultures. The system of cyclic secondary somatic embryogenesis in black pepper described here represents a permanent source of embryogenic material that can be used for genetic manipulations of this crop species.

  6. In vitro study on regeneration of Gladiolus grandiflorus corm calli as affected by plant growth regulators.

    Science.gov (United States)

    Torabi-Giglou, Mousa; Hajieghrari, Behzad

    2008-04-15

    In this study, in vitro organogenesis of Gladiolus grandiflorus cultivar pink corm segments were evaluated by culturing corm calli in modified MS medium supplemented with 3% sucrose and 0.7% agar with different concentration of BAP (0, 1, 2 and 4 mg L(-1) medium) and NAA (0, 0.5, 1 and 2 mg L(-1) medium) in factorial experiment of Completely Randomized Design (CRD). In order to obtain Gladiolus calli, corm segments (Aprox. 5 x 5 x 1 mm in size) were kept in modified MS medium (Murashige and Skoog, 1962) that was supplemented with 1 mg L(-1) 2, 4-D, 3% sucrose and 0.7% agar. The results showed that increasing the concentration of BAP from 0 to 2 mg L(-1) medium simulated plantlet regeneration but no significantly effect was obtained on shoot and cormel organogenesis between 2 and 4 mg L(-1) BAP concentration in medium. Increasing of NAA content in media without BAP developed rootlet significantly. Interaction results showed that increasing BAP content against decreasing of NAA concentration stimulates the shoot and cormel proliferation.

  7. Thidiazuron promotes high frequency regeneration of peanut (Arachis hypogaea) plants in vitro.

    Science.gov (United States)

    Kanyand, M; Dessai, A P; Prakash, C S

    1994-11-01

    Multiple shoots were induced on Valenciatype peanut (Arachis hypogaea L.) explants cultured in vitro on a nutrient medium supplemented with thidiazuron. Zygotic embryos excised from mature seeds were germinated on Murashige-Skoog nutrient medium, and the resulting plantlets (8 days-old) were used as a source of explants. When cultured on a nutrient medium with increasing levels of thidiazuron (0.5 to 30 mg/l), expiants from various parts of the peanut plant (except the root) produced multiple shoot primordia which subsequently developed into individual shoots. Hypocotyl and cotyledon explants produced shoots in higher numbers than other explants (20 shoots per hypocotyl explant at all thidiazuron concentrations and 15 shoots per cotyledon explant at 30 mg/l). Shoots rooted normally on a basal Murashige-Skoog medium containing charcoal and developed into healthy and fertile plants when planted in soil.

  8. Seedling Survival and Natural Regeneration For a Bottomland Hardwood Planting on Sites Differing in Site Preparation

    Science.gov (United States)

    Daniel T. Johns; Brett Williams; Hans M. Williams; Matthew Stroupe

    1999-01-01

    In January 1998, three tracts in Hardin County, TX, were hand-planted with seven species of 1-0 bareroot bottomland hardwood seedlings. The tracts, managed by The Nature Conservancy of Texas, were previously 20-year-old pine plantations. The tracts are located within the floodplain of Village Creek. An objective for this conversion is the restoration of a bottomland...

  9. In vitro plant regeneration from protocorms-like bodies (PLBs) and ...

    African Journals Online (AJOL)

    Phalaenopsis, with long arching sprays of flowers, are among the most beautiful flowers in the world. Phalaenopsis is an important genus and one of the most popular epiphytic monopodial orchids, grown commercially for the production of cut flowers and potted plants. Most of them have different and interesting ...

  10. An efficient and reproducible method for regeneration of whole plants from mature seeds of a high yielding Indica rice (Oryza sativa L.) variety PAU 201.

    Science.gov (United States)

    Wani, Shabir H; Sanghera, Gulzar S; Gosal, Satbir S

    2011-07-01

    Tissue culture is one of the tools necessary for genetic engineering and many other breeding programs. Moreover, selection of high regenerating rice varieties is a pre-requisite for success in rice biotechnology. In this report we established a reproducible plant regeneration system through somatic embryogenesis. The explants used for regeneration were embryogenic calli derived from mature seeds cultured on callus induction media. For callus induction mature seeds were cultured on MS medium containing 30 g/l sucrose combined with 560 mg/l proline and 1.5-3.5 mg/l 2,4-D and 0.5-1.5 mg/l Kin. For plant regeneration, embryogenic calli were transferred to MS medium containing 30 g/l sucrose, supplemented with 1.0-3.0 mg/l BAP, 0.5-1.5 mg/l Kin and 0.5-1.5 mg/l NAA. The highest frequency of callus induction (44.4%) was observed on the MS medium supplemented with 2.5 mg/l 2,4-D, 0.5 mg/l Kin, 560 mg/l proline and 30 g/l sucrose. The highest frequency of shoot regeneration (42.5%) was observed on the MS medium supplemented with 2.0 mg/l BAP, 0.5 mg/l NAA and 0.5 mg/l Kin. The plantlets were hardened and transferred to soil in earthen pots. The developed method was highly reproducible. The in vitro developed plants showed normal growth and flowering under glasshouse conditions. Copyright © 2011 Elsevier B.V. All rights reserved.

  11. C/sub 4/ photosynthesis in Euphorbia degeneri and E. remyi: a comparison of photosynthetic carbon metabolism in leaves, callus cultures and regenerated plants

    Energy Technology Data Exchange (ETDEWEB)

    Ruzin, S.E.

    1984-04-01

    Based on analysis of /sup 14/CO/sub 2/ fixation kinetics and assays of enzymes related to C/sub 4/ metabolism (NAD-ME, NADP-ME, NAD-MDH, NADP-MDH, AST, ALT), leaves and regenerated plants of Euphorbia degeneri exhibit a modified NADP-ME-type photosynthesis. Apparently, both aspartate and malate are used for transport of CO/sub 2/ to bundle sheath cells. Callus grown on either non-shoot-forming or shoot-forming media fixes CO/sub 2/ into RPP-cycle intermediates and sucrose, as well as malate and aspartate. /sup 14/CO/sub 2/ pulse/chase kinetics show no significant loss of label from C/sub 4/ acids throughout a one minute chase. Analysis of PEPCase revealed the presence of 2 isoenzymes in both leaf and regenerated plant tissues (K/sub m/ (PEP) = 0.080 and 0.550) but only one isoenzyme in callus (K/sub m/ = 0.100). It appears that C/sub 4/ photosynthesis does not occur in callus derived from this C/sub 4/ dicot but is regenerated concomitant with shoot regeneration, and ..beta..-carboxylation of PEP in callus, mediated by the low K/sub m/ isoenzyme of PEPCase, produces C/sub 4/ acids that are not involved in the CO/sub 2/ shuttle mechanism characteristic of C/sub 4/ photosynthesis. 161 references, 19 figures, 12 tables.

  12. Loss of CMD2?mediated resistance to cassava mosaic disease in plants regenerated through somatic embryogenesis

    OpenAIRE

    Beyene, Getu; Chauhan, Raj Deepika; Wagaba, Henry; Moll, Theodore; Alicai, Titus; Miano, Douglas; Carrington, James C.; Taylor, Nigel J.

    2016-01-01

    Summary Cassava mosaic disease (CMD) and cassava brown streak disease (CBSD) are the two most important viral diseases affecting cassava production in Africa. Three sources of resistance are employed to combat CMD: polygenic recessive resistance, termed CMD1, the dominant monogenic type, named CMD2, and the recently characterized CMD3. The farmer?preferred cultivar TME 204 carries inherent resistance to CMD mediated by CMD2, but is highly susceptible to CBSD. Selected plants of TME 204 produc...

  13. The Effect of Plant Growth Regulators on Callus Induction and Regeneration of Amygdalus communis

    Directory of Open Access Journals (Sweden)

    Naimeh SHARIFMOGHADAM

    2011-08-01

    Full Text Available The Almond (Amygdalus communis is one of the most important and oldest commercial nut crops, belonging to the Rosaceae family. Almond has been used as base material in pharmaceutical, cosmetic, hygienically and food industry. Propagation by tissue culture technique is the most important one in woody plants. In the current research, in vitro optimization of tissue culture and mass production of almond was investigated. In this idea, explants of actively growing shoots were collected and sterilized, then transferred to MS medium with different concentrations and combinations of plant growth regulators. The experiment was done in completely randomized blocks design, with 7 treatment and 30 replications. After 4 weeks, calli induction, proliferation, shoot length and number of shoot per explants were measured. Results showed that the best medium for shoot initiation and proliferation was MS + 0.5 mg/l IAA (Indol-3-Acetic Acid + 1 mg/l BA (Benzyl Adenine. Autumn was the best season for collecting explants. The shoots were transferred to root induction medium with different concentrations of plant growth regulators. The best root induction medium was MS + 0.5 mg/l IBA (Indol Butyric Acid.

  14. Loop-mediated Isothermal Amplification Assay to Rapidly Detect Wheat Streak Mosaic Virus in Quarantined Plants

    Directory of Open Access Journals (Sweden)

    Siwon Lee

    2015-12-01

    Full Text Available We developed a loop-mediated isothermal amplification (LAMP method to rapidly diagnose Wheat streak mosaic virus (WSMV during quarantine inspections of imported wheat, corn, oats, and millet. The LAMP method was developed as a plant quarantine inspection method for the first time, and its simplicity, quickness, specificity and sensitivity were verified compared to current reverse transcription-polymerase chain reaction (RT-PCR and nested PCR quarantine methods. We were able to quickly screen for WSMV at quarantine sites with many test samples; thus, this method is expected to contribute to plant quarantine inspections.

  15. In vitro propagation and genetic fidelity study of plant regenerated from inverted hypocotyl explants of eggplant (Solanum melongena L.) cv. Arka Shirish.

    Science.gov (United States)

    Padma Mallaya, N; Ravishankar, G A

    2013-02-01

    Genetic variation due to somaclonal variation in micropropagated plants is a beneficial phenomenon for crop improvement. Genetic integrity of the plants derived through micropropagation becomes crucial if genetic transformation studies have to be carried out. Somaclonal variation in tissue culture is a common phenomenon which makes it mandatory to check for genetic stability of plants. Hypocotyl explants of Solanummelongena L. cv. Arka Shirish inoculated with inverted polarity in MS media supplemented with 0.5 mg L(-1) thidiazuron (TDZ) gave maximum number of shoot buds. Elongation of the shoot buds was achieved on MS medium supplemented with 0.5 mg L(-1) 2, 3, 5-triiodobenzoic acid (TIBA) and 0.1 mg L(-1) gibberellic acid (GA3). The elongated shoots were rooted in MS with 1 mg L(-1) indole-3-butyric acid (IBA), and the rooted plants were hardened in the greenhouse. Morphological characteristics were similar in both seed-propagated and micropropagated plants. Random amplified polymorphic DNA analysis carried out with 10 primers for genetic stability studies of the regenerated plants generated 96 scorable bands with a total of 1,056 bands for the primers. Comparison of the bands with the mother plant revealed the monomorphic nature and true-to-type clones. The above regeneration protocol will be useful for micropropagation and genetic transformation studies of S.melongena L. cv. Arka Shirish.

  16. Fungal phytopathogens encode functional homologues of plant rapid alkalinization factor (RALF) peptides.

    Science.gov (United States)

    Thynne, Elisha; Saur, Isabel M L; Simbaqueba, Jaime; Ogilvie, Huw A; Gonzalez-Cendales, Yvonne; Mead, Oliver; Taranto, Adam; Catanzariti, Ann-Maree; McDonald, Megan C; Schwessinger, Benjamin; Jones, David A; Rathjen, John P; Solomon, Peter S

    2017-08-01

    In this article, we describe the presence of genes encoding close homologues of an endogenous plant peptide, rapid alkalinization factor (RALF), within the genomes of 26 species of phytopathogenic fungi. Members of the RALF family are key growth factors in plants, and the sequence of the RALF active region is well conserved between plant and fungal proteins. RALF1-like sequences were observed in most cases; however, RALF27-like sequences were present in the Sphaerulina musiva and Septoria populicola genomes. These two species are pathogens of poplar and, interestingly, the closest relative to their respective RALF genes is a poplar RALF27-like sequence. RALF peptides control cellular expansion during plant development, but were originally defined on the basis of their ability to induce rapid alkalinization in tobacco cell cultures. To test whether the fungal RALF peptides were biologically active in plants, we synthesized RALF peptides corresponding to those encoded by two sequenced genomes of the tomato pathogen Fusarium oxysporum f. sp. lycopersici. One of these peptides inhibited the growth of tomato seedlings and elicited responses in tomato and Nicotiana benthamiana typical of endogenous plant RALF peptides (reactive oxygen species burst, induced alkalinization and mitogen-activated protein kinase activation). Gene expression analysis confirmed that a RALF-encoding gene in F. oxysporum f. sp. lycopersici was expressed during infection on tomato. However, a subsequent reverse genetics approach revealed that the RALF peptide was not required by F. oxysporum f. sp. lycopersici for infection on tomato roots. This study has demonstrated the presence of functionally active RALF peptides encoded within phytopathogens that harbour an as yet undetermined role in plant-pathogen interactions. © 2016 BSPP AND JOHN WILEY & SONS LTD.

  17. Regeneration of plants from leaves of Chrysanthemum morifolium Ram. cv. Bronze Bornholm in in vitro cultures

    Directory of Open Access Journals (Sweden)

    Aurelia Ślusarkiewicz-Jarzina

    2014-01-01

    Full Text Available Plants were obtained from cultured in vitro leaves of Chrysanthemum morifolium Ram. cv. Bronze Bornholm. The leaves were inoculated on Murashige and Skoog medium (MS supplemented with cytokinins (kinetin - KIN, zeatin - ZEA, 6-benzyloaminopurine - BAP and auxins (2,4-dichlorophenoxyacetic acid - 2,4-D, α-naphtaleneacetic acid - NAA, 3-indolilacetic acid - IAA, p-fluorophenylalanine - PFA in various combinations and concentra-tions. The most suitable medium was that one which contained 4 mg/l KIN, 2 mg/l NAA and 50 mg/l PFA.

  18. CALLUS INDUCTION AND PLANT REGENERATION IN PUNICA GRANATUM L. ?NANA' FROM LEAF EXPLANTS

    Directory of Open Access Journals (Sweden)

    Alireza Bonyanpour

    2013-09-01

    Full Text Available ABSTRACT In this investigation, leaf explants of a local cultivar of dwarf pomegranate were placed on Murashige and Skoog (1962 (MS medium supplemented with various concentrations of 6-benzyl adenin (BA and naphthalene acetic acid (NAA for callus induction. After 40 days, maximum callus induction was observed on a media containing 1 mg L-1 BA and 0.2 to 0.4 mg L-1 NAA. However, the highest callus growth was obtained on a medium containing 1 mg L-1 BA and 1 mg L-1 NAA. The highest number of shoots (7 shoots per explants was obtained by transferring the calli to the media containing 5 mg L-1 BA with 0.1 mg L-1 NAA. Maximum shoot proliferation was observed when shoots were cultured on woody plant medium (WPM supplemented with 5 mg L-1 kinetin (Kin. In this treatment, after 4 subcultures, 36 shoots were produced from one original explant. Among treatments used in rooting experiments, shoots cultured on WPM medium containing 0.2 mg L-1 indol butyric acid (IBA had the maximum root percentage (100% and good root growth (2.06 cm mean length and 2 roots in each explants. Rooted plantlets were cultured in a soil mixture containing vermiculite (60%, perlite (30% and coco peat (10% v/v. After 2 months, 80% of plants survived and transferred to the greenhouse.

  19. Development of Rapid Isothermal Amplification Assays for Detection of Phytophthora spp. in Plant Tissue.

    Science.gov (United States)

    Miles, Timothy D; Martin, Frank N; Coffey, Michael D

    2015-02-01

    Several isothermal amplification techniques recently have been developed that are tolerant of inhibitors present in many plant extracts, which can reduce the need for obtaining purified DNA for running diagnostic assays. One such commercially available technique that has similarities with real-time polymerase chain reaction (PCR) for designing primers and a labeled probe is recombinase polymerase amplification (RPA). This technology was used to develop two simple and rapid approaches for detection of Phytophthora spp.: one genus-specific assay multiplexed with a plant internal control and the other species-specific assays for Phytophthora ramorum and P. kernoviae. All assays were tested for sensitivity (ranging from 3 ng to 1 fg of DNA) and specificity using DNA extracted from more than 136 Phytophthora taxa, 21 Pythium spp., 1 Phytopythium sp., and a wide range of plant species. The lower limit of linear detection using purified DNA was 200 to 300 fg of DNA in all pathogen RPA assays. Six different extraction buffers were tested for use during plant tissue maceration and the assays were validated in the field by collecting 222 symptomatic plant samples from over 50 different hosts. Only 56 samples were culture positive for Phytophthora spp. whereas 91 were positive using the Phytophthora genus-specific RPA test and a TaqMan real-time PCR assay. A technique for the generation of sequencing templates from positive RPA amplifications to confirm species identification was also developed. These RPA assays have added benefits over traditional technologies because they are rapid (results can be obtained in as little as 15 min), do not require DNA extraction or extensive training to complete, use less expensive portable equipment than PCR-based assays, and are significantly more specific than current immunologically based methods. This should provide a rapid, field-deployable capability for pathogen detection that will facilitate point-of-sample collection processing

  20. The practical influence of rapid mixing on coagulation in a full-scale water treatment plant.

    Science.gov (United States)

    Allerdings, Demitri; Förster, Gerrit; Vasyukova, Ekaterina; Uhl, Wolfgang

    2015-01-01

    This study focuses on the effect of rapid mixing on the coagulation efficiency in a full-scale drinking-water treatment plant and discusses the mechanisms involved in the floc-formation process. The results refer to three periods of operation of the waterworks when no mechanical mixing was provided in the tanks for coagulant dosing due to mechanical failure of the rapid mixers. Although a certain deterioration of the subsequent flocculation process was observed, as assessed using the data for suspended solids, turbidity, and chemical oxygen demand, the overall water treatment performance was not affected. This suggests an insignificant role for intense rapid mixing in sweep flocculation during full-scale water treatment and reveals the potential to reduce the required energy costs for mechanical mixers.

  1. Effects of Different Media, Type and Concentrations of Auxin on Callus Induction and Plant Regeneration from Young Inflorescences of Yellow Bluestem (Bothriochloa ischaemum(L.) Keng)

    OpenAIRE

    CAN, Ersin

    2000-01-01

    This study was conducted to determine the effects of different media, auxin types and concentrations on callus induction and plant regeneration from young inflorescences of 15 ecotypes of yellow bluestem ( Bothriochloa ischaemum). The young inflorescences were cultured on two different basal media (LS and SH) containing different concentrations (2, 4, 6, and 8 mg/l) of two different auxins (2, 4-D and Dicamba). The results showed that the ecotypes were significantly different from each ot...

  2. The impact of Cu treatment on phenolic and polyamine levels in plant material regenerated from embryos obtained in anther culture of carrot

    Czech Academy of Sciences Publication Activity Database

    Górecka, K.; Cvikrová, Milena; Kowalska, U.; Eder, Josef; Szafrańska, K.; Górecki, R.; Janas, K. M.

    2007-01-01

    Roč. 45, č. 1 (2007), s. 54-61 ISSN 0981-9428 R&D Projects: GA MŠk 1P05OC052 Institutional research plan: CEZ:AV0Z50380511 Source of funding: V - iné verejné zdroje Keywords : carrot culture * copper ions * embryo regeneration Subject RIV: GE - Plant Breeding Impact factor: 1.669, year: 2007

  3. A simple electrochemical method for the rapid estimation of antioxidant potentials of some selected medicinal plants.

    Science.gov (United States)

    Amidi, Salimeh; Mojab, Faraz; Bayandori Moghaddam, Abdolmajid; Tabib, Kimia; Kobarfard, Farzad

    2012-01-01

    Clinical and Epidemiological studies have shown that a diet rich in fruits and vegetables is associated with a decreased risk of cardiovascular diseases, cancers and other related disorders. These beneficial health effects have been attributed in part to the presence of antioxidants in dietary plants. Therefore screening for antioxidant properties of plant extracts has been one of the interests of scientists in this field. Different screening methods have been reported for the evaluation of antioxidant properties of plant extracts in the literature. In the present research a rapid screening method has been introduced based on cyclic voltammetry for antioxidant screening of some selected medicinal plant extracts. CYCLIC VOLTAMMETRY OF METHANOLIC EXTRACTS OF SEVEN MEDICINAL PLANTS: Buxus hyrcana, Rumex crispus, Achillea millefolium, Zataria multiflora, Ginkgo biloba, Lippia citriodora and Heptaptera anisoptera was carried out at different scan rates. Based on the interpretation of voltammograms, Rumex crispus, Achillea millefolium and Ginkgo biloba showed higher antioxidant capability than the others while Lippia citriodora contained the highest amount of antioxidants. Cyclic voltammetry is expected to be a simple method for screening antioxidants and estimating the antioxidant activity of foods and medicinal plants.

  4. Conversion of oat (Avena sativaL.) haploid embryos into plants in relation to embryo developmental stage and regeneration media.

    Science.gov (United States)

    Noga, Angelika; Skrzypek, Edyta; Warchoł, Marzena; Czyczyło-Mysza, Ilona; Dziurka, Kinga; Marcińska, Izabela; Juzoń, Katarzyna; Warzecha, Tomasz; Sutkowska, Agnieszka; Nita, Zygmunt; Werwińska, Krystyna

    2016-01-01

    Obtaining oat DH lines is only effective via wide crossing with maize. Seven hundred haploid embryos from 21 single F 1 progeny obtained from wide crosses with maize were isolated, divided into four groups according to their size (<0.5 mm, 0.5-0.9 mm, 1.0-1.4 mm, and ≥1.5 mm), and transferred into 190-2 regeneration medium with different growth regulators: 0.5 mg L -1 kinetin (KIN) and 0.5 mg L -1 1-naphthaleneacetic acid (NAA); 1 mg L -1 zeatin (ZEA) and 0.5 mg L -1 NAA; or 1 mg L -1 dicamba (DIC), 1 mg L -1 picloram (PIC), and 0.5 mg L -1 kinetin (KIN). Among all isolated embryos, approximately 46.1% were between 1.0-1.4 mm, while the smallest group of embryos (7.1%) were those <0.5 mm. The ability of haploid embryos to germinate varied depending on oat genotypes and the size of embryos. Haploid embryos <0.5 mm were globular and did not germinate, whereas embryos ≥1.5 mm had clearly visible coleoptiles, radicles, and scutella, and were able to germinate. Germination of oat haploid embryos varied depending on growth regulators in the regeneration medium. Most haploid embryos germinated on medium with 0.5 mg L -1 NAA and 0.5 mg L -1 KIN, while the fewest germinated on medium with 1 mg L -1 DIC, 1 mg L -1 PIC, and 0.5 mg L -1 KIN. One hundred thirty germinated haploid embryos converted into haploid plants. Fifty oat DH lines were obtained after colchicine treatment.

  5. High-efficiency Agrobacterium-mediated transformation of chickpea (Cicer arietinum L.) and regeneration of insect-resistant transgenic plants.

    Science.gov (United States)

    Mehrotra, Meenakshi; Sanyal, Indraneel; Amla, D V

    2011-09-01

    To develop an efficient genetic transformation system of chickpea (Cicer arietinum L.), callus derived from mature embryonic axes of variety P-362 was transformed with Agrobacterium tumefaciens strain LBA4404 harboring p35SGUS-INT plasmid containing the uidA gene encoding β-glucuronidase (GUS) and the nptII gene for kanamycin selection. Various factors affecting transformation efficiency were optimized; as Agrobacterium suspension at OD(600) 0.3 with 48 h of co-cultivation period at 20°C was found optimal for transforming 10-day-old MEA-derived callus. Inclusion of 200 μM acetosyringone, sonication for 4 s with vacuum infiltration for 6 min improved the number of GUS foci per responding explant from 1.0 to 38.6, as determined by histochemical GUS assay. For introducing the insect-resistant trait into chickpea, binary vector pRD400-cry1Ac was also transformed under optimized conditions and 18 T(0) transgenic plants were generated, representing 3.6% transformation frequency. T(0) transgenic plants reflected Mendelian inheritance pattern of transgene segregation in T(1) progeny. PCR, RT-PCR, and Southern hybridization analysis of T(0) and T(1) transgenic plants confirmed stable integration of transgenes into the chickpea genome. The expression level of Bt-Cry protein in T(0) and T(1) transgenic chickpea plants was achieved maximum up to 116 ng mg(-1) of soluble protein, which efficiently causes 100% mortality to second instar larvae of Helicoverpa armigera as analyzed by an insect mortality bioassay. Our results demonstrate an efficient and rapid transformation system of chickpea for producing non-chimeric transgenic plants with high frequency. These findings will certainly accelerate the development of chickpea plants with novel traits.

  6. Fabrication of novel Si-doped hydroxyapatite/gelatine scaffolds by rapid prototyping for drug delivery and bone regeneration.

    Science.gov (United States)

    Martínez-Vázquez, F J; Cabañas, M V; Paris, J L; Lozano, D; Vallet-Regí, M

    2015-03-01

    Porous 3-D scaffolds consisting of gelatine and Si-doped hydroxyapatite were fabricated at room temperature by rapid prototyping. Microscopic characterization revealed a highly homogeneous structure, showing the pre-designed porosity (macroporosity) and a lesser in-rod porosity (microporosity). The mechanical properties of such scaffolds are close to those of trabecular bone of the same density. The biological behavior of these hybrid scaffolds is greater than that of pure ceramic scaffolds without gelatine, increasing pre-osteoblastic MC3T3-E1 cell differentiation (matrix mineralization and gene expression). Since the fabrication process of these structures was carried out at mild conditions, an antibiotic (vancomycin) was incorporated in the slurry before the extrusion of the structures. The release profile of this antibiotic was measured in phosphate-buffered saline solution by high-performance liquid chromatography and was adjusted to a first-order release kinetics. Vancomycin released from the material was also shown to inhibit bacterial growth in vitro. The implications of these results for bone tissue engineering applications are discussed. Copyright © 2014 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  7. Agrobacterium-mediated genetic transformation and regeneration of transgenic plants using leaf midribs as explants in ramie [Boehmeria nivea (L.) Gaud].

    Science.gov (United States)

    An, Xia; Wang, Bo; Liu, Lijun; Jiang, Hui; Chen, Jie; Ye, Shengtuo; Chen, Leiyu; Guo, Pingan; Huang, Xing; Peng, Dingxiang

    2014-05-01

    In this study, leaf midribs, the elite explants, were used for the first time to develop an efficient regeneration and transformation protocol for ramie [Boehmeria nivea (L.) Gaud.] via Agrobacterium-mediated genetic transformation. Sensitivity of leaf midribs regeneration to kanamycin was evaluated, which showed that 40 mg l(-1) was the optimal concentration needed to create the necessary selection pressure. Factors affecting the ramie transformation efficiency were evaluated, including leaf age, Agrobacterium concentration, length of infection time for the Agrobacterium solution, acetosyringone concentration in the co-cultivation medium, and the co-cultivation period. The midrib explants from 40-day-old in vitro shoots, an Agrobacterium concentration at OD600 of 0.6, 10-min immersion in the bacteria solution, an acetosyringone concentration of 50 mg l(-1) in the co-cultivation medium and a 3-day co-cultivation period produced the highest efficiencies of regeneration and transformation. In this study, the average transformation rate was 23.25%. Polymerase chain reactions using GUS and NPTII gene-specific primers, Southern blot and histochemical GUS staining analyses further confirmed that the transgene was integrated into the ramie genome and expressed in the transgenic ramie. The establishment of this system of Agrobacterium-mediated genetic transformation and regeneration of transgenic plants will be used not only to introduce genes of interest into the ramie genome for the purpose of trait improvement, but also as a common means of testing gene function by enhancing or inhibiting the expression of target genes.

  8. Efficient plant regeneration protocol through callus for Saussurea obvallata (DC.) Edgew. (Asteraceae): effect of explant type, age and plant growth regulators.

    Science.gov (United States)

    Dhar, Uppeandra; Joshi, Mitali

    2005-06-01

    A callus induction and in vitro plantlet regeneration system for the endangered state flower of Uttaranchal (Saussurea obvallata) was optimized by studying the influence of explant type (root, hypocotyl, cotyledon and leaf), age and different concentrations of plant growth regulators. Explants from 10 to 15-day-old seedlings showed maximum callus induction. Callus formation and shoot differentiation was initiated on Murashige-Skoog (MS) medium containing 6-benzyladenine (BA) and alpha-naphthalene acetic acid (NAA) in all explant types. The best results were obtained using leaf explants: 100% callusing was achieved in MS medium supplemented with 2.5 microM BA and 1.0 microM NAA, and 100% differentiation along with a multiplication rate of 12 shoots per explant with a combination of 5.0 microM BA and 1.0 microM NAA. However, the results reflected the existence of high inter-explant variability in response to growth regulators. In vitro rooting of shoots was achieved at an efficiency of 100% in one-half strength MS medium supplemented with 2.5 microM indole-3-butyric acid. Application of this protocol has potential for mass multiplication of the target species in a limited time period.

  9. Protocol: a rapid and economical procedure for purification of plasmid or plant DNA with diverse applications in plant biology

    Directory of Open Access Journals (Sweden)

    Li Li

    2010-01-01

    Full Text Available Abstract Research in plant molecular biology involves DNA purification on a daily basis. Although different commercial kits enable convenient extraction of high-quality DNA from E. coli cells, PCR and agarose gel samples as well as plant tissues, each kit is designed for a particular type of DNA extraction work, and the cost of purchasing these kits over a long run can be considerable. Furthermore, a simple method for the isolation of binary plasmid from Agrobacterium tumefaciens cells with satisfactory yield is lacking. Here we describe an easy protocol using homemade silicon dioxide matrix and seven simple solutions for DNA extraction from E. coli and A. tumefaciens cells, PCR and restriction digests, agarose gel slices, and plant tissues. Compared with the commercial kits, this protocol allows rapid DNA purification from diverse sources with comparable yield and purity at negligible cost. Following this protocol, we have demonstrated: (1 DNA fragments as small as a MYC-epitope tag coding sequence can be successfully recovered from an agarose gel slice; (2 Miniprep DNA from E. coli can be eluted with as little as 5 μl water, leading to high DNA concentrations (>1 μg/μl for efficient biolistic bombardment of Arabidopsis seedlings, polyethylene glycol (PEG-mediated Arabidopsis protoplast transfection and maize protoplast electroporation; (3 Binary plasmid DNA prepared from A. tumefaciens is suitable for verification by restriction analysis without the need for large scale propagation; (4 High-quality genomic DNA is readily isolated from several plant species including Arabidopsis, tobacco and maize. Thus, the silicon dioxide matrix-based DNA purification protocol offers an easy, efficient and economical way to extract DNA for various purposes in plant research.

  10. Callus formation and plant regeneration from protoplasts of sunflower calli and hypocotyls

    Directory of Open Access Journals (Sweden)

    Conceição Santos

    2014-01-01

    Full Text Available Sunflower (cv. Girapac SH222 protoplasts were obtained from 4-7 day-old hypocotyls and cotyledons and from two-month old calli. Higher yields of protoplasts were achieved with medium El (KCl 25g dm-3, CaCl2 2g dm-3, MES 0.7 g• dm-3, pH 5.5 and the combination of Driselase Fluka 0.2%, Macerozyme Onozuka 0.2% and Cellulase Onozuka R10 0.2%. Hypocotyls gave the highest yields of protoplasts, followed by cotyledons and calli. Protoplasts were cultivated in liquid and on solid media using both L4M (Burrus et al., 1991 and V-KM (Bokelman and Roest, 1983 media. Culture on solid M1 medium (L4M medium supplemented with NAA 3.0 mg•dm- 3, 2,4-D 0.1 mg•dm-3 and BA 1.0 mg•dm -3 gave a good planting efficiency with the development of many white-green colonies. These colonies gave rise to small calli which were transferred to MSmod medium (MS medium supplemented with KCI 5 g•dm-', and polyvinylpyrrolidone (PVP, 4 g• dm-3 containing benziladenine (BA, 0.5 mg•dm-3, naphtaleneacetic acid (NAA, 0.5 mg•dm-3 and giberelic acid (GA3 0.1 mg•dm-3. After two weeks, calli were transferred to MSmod medium containing BA 1.0 mg•dm-3, NAA 0.1 mg•dm-3, and GA3 0.1 mg•dm-3 for shoot formation. Shoots were excised and induced to root in MSmod supplemented with BA 0.1 mg•dm-3, NAA 1.0 mg•dm-3, and GA3 0.1 mg•dm-3. Plantlets were then transferred to sterilised vermiculite for greenhouse acclimation.

  11. REGENERATION OF LEACHATES AFTER SULPHATE COOKINGS OF PLANT RAW MATERIALS IN PRESENCE OF METAL SALT WITH TRANSITION VALENCY

    Directory of Open Access Journals (Sweden)

    I. Karpunin

    2013-01-01

    Full Text Available The paper proposes schemes for introduction and regeneration of added salt during wood alkaline pulping under industrial conditions. Reactions proceeding with blue copper and leachates at their burning have been determined in the paper.

  12. Rapid in vitro propagation system through shoot tip cultures of Vitex trifolia L.-an important multipurpose plant of the Pacific traditional Medicine.

    Science.gov (United States)

    Ahmed, Rafique; Anis, Mohammad

    2014-07-01

    A rapid and efficient plant propagation system through shoot tip explants was established in Vitex trifolia L., a medicinally important plant belonging to the family Verbenaceae. Multiple shoots were induced directly on Murashige and Skoog (MS) medium consisting of different cytokinins, 6-benzyladenine (BA), kinetin (Kin) and 2-isopentenyl adenine (2-iP), BA at an optimal concentration of 5.0 μM was most effective in inducing multiple shoots where 90 % explants responded with an average shoot number (4.4±0.1) and shoot length (2.0±0.1 cm) after 6 weeks of culture. Inclusion of NAA in the culture medium along with the optimum concentration of BA promoted a higher rate of shoot multiplication and length of the shoot, where 19.2±0.3 well-grown healthy shoots with an average shoot length of 4.4±0.1 cm were obtained on completion of 12 weeks culture period. Ex vitro rooting was achieved best directly in soilrite when basal portion of the shoots were treated with 500 μM indole-3-butyric acid for 15 min which was the most effective in inducing roots, as 95 % of the microshoots produced roots. Plantlets went through a hardening phase in a controlled plant growth chamber, prior to ex-vitro transfer. Micropropagated plants grew well, attained maturity and flowered with 92 % survival rate. The results of this study provide the first report on in vitro plant regeneration of Vitex trifolia L. using shoot tip explants.

  13. High frequency microcloning of Aloe vera and their true-to-type conformity by molecular cytogenetic assessment of two years old field growing regenerated plants.

    Science.gov (United States)

    Haque, Sk Moquammel; Ghosh, Biswajit

    2013-12-01

    Aloe vera (L.) Burm.f is an important industrial crop, which has enormous application in pharmaceutical, cosmetic and food industries. Thereby, the demand for quality planting material of A. vera is increasing worldwide. Micropropagation is the widely accepted practical application of plant biotechnology that has gained the status of a multibillion-dollar industry throughout the world and this techniques can be used to meet the industrial demand of A. vera. Present studies aim to develop a proficient methods of high-frequency true-to-type plantlet regeneration without intermediate callus phase for A. vera. Nodal portion of rhizomatous stem of A. vera were cultured on Murashige and Skoog (MS) medium (Physiol. Plant. 15:473 - 497, 1962) supplemented with various cytokinin and A. vera leaf gel (AvG) as organic supplement. Number of proliferated shoots per explant was increased along with the regeneration cycles and on MS medium supplemented with 2.5 mg/L 6-benzylaminopurine and 10.0% (v/v) AvG, only 17.8 ± 0.35 shoots per explant were induced on 1(st) regeneration cycle whereas on 3(rd) regeneration cycle these number increase to 38.5 ± 0.44 shoots per explant on the same medium composition. AvG have an encouraging role to increase the proliferation rate and on 3(rd) regeneration cycle 27.6 ± 0.53 shoot per explant induced on 2.5 mg/L BAP, but these number increase to 38.5 ± 0.44 shoots per explant when 10.0% (v/v) AvG was added along with 2.5 mg/L BAP. After transfer of individual excised shoots to a one-third strength MS medium containing 20.0% (v/v) AvG, all the shoots formed whole plantlets with maximum number (9.6 ± 0.29) of roots per shoot. 95.0% of the regenerated plantlets survived on poly-green house. Normal flower appeared in 84.2% field growing micropropagated plants after 18 to 20 months of field transfer. Further, clonal fidelity of the two years old micropropagated plants was established by studying mitotic and meiotic

  14. Automated electrical impedance technique for rapid enumeration of fecal coliforms in effluents from sewage treatment plants.

    Science.gov (United States)

    Silverman, M P; Munoz, E F

    1979-01-01

    Fecal coliforms growing in a selective lactose-based broth medium at 44.5 degrees C generate a change in the electrical impedance of the culture relative to a sterile control when populations reach 10(6) to 10(7) per ml. The ratio of these changes was measured automatically, and the data were processed by computer. A linear relation was found between the log10 of the number of fecal coliforms in an inoculum and the time required for an electrical impedance ratio signal to be detected. Pure culture inocula consisting of 100 fecal coliforms in log phase or stationary phase were detected in 6.5 and 7.7 h, respectively. Standard curves of log10 fecal coliforms in wastewater inocula versus detection time, based on samples collected at a sewage treatment plant over a 4-month period, were found to vary from one another with time. Nevertheless, detection times were rapid and ranged from 5.8 to 7.9 h for 200 fecal coliforms to 8.7 to 11.4 h for 1 fecal coliform. Variations in detection times for a given number of fecal coliforms were also found among sewage treatment plants. A strategy is proposed which takes these variations into account and allows for rapid, automated enumeration of fecal coliforms in wastewater by the electrical impedance ratio technique. PMID:378128

  15. Rapid in vivo analysis of synthetic promoters for plant pathogen phytosensing

    Directory of Open Access Journals (Sweden)

    Liu Wusheng

    2011-11-01

    Full Text Available Abstract Background We aimed to engineer transgenic plants for the purpose of early detection of plant pathogen infection, which was accomplished by employing synthetic pathogen inducible promoters fused to reporter genes for altered phenotypes in response to the pathogen infection. Toward this end, a number of synthetic promoters consisting of inducible regulatory elements fused to a red fluorescent protein (RFP reporter were constructed for use in phytosensing. Results For rapid analysis, an Agrobacterium-mediated transient expression assay was evaluated, then utilized to assess the inducibility of each synthetic promoter construct in vivo. Tobacco (Nicotiana tabacum cv. Xanthi leaves were infiltrated with Agrobacterium harboring the individual synthetic promoter-reporter constructs. The infiltrated tobacco leaves were re-infiltrated with biotic (bacterial pathogens or abiotic (plant defense signal molecules salicylic acid, ethylene and methyl jasmonate agents 24 and 48 hours after initial agroinfiltration, followed by RFP measurements at relevant time points after treatment. These analyses indicated that the synthetic promoter constructs were capable of conferring the inducibility of the RFP reporter in response to appropriate phytohormones and bacterial pathogens, accordingly. Conclusions These observations demonstrate that the Agrobacterium-mediated transient expression is an efficient method for in vivo assays of promoter constructs in less than one week. Our results provide the opportunity to gain further insights into the versatility of the expression system as a potential tool for high-throughput in planta expression screening prior to generating stably transgenic plants for pathogen phytosensing. This system could also be utilized for temporary phytosensing; e.g., not requiring stably transgenic plants.

  16. A Method to Teach Age-Specific Demography with Field Grown Rapid Cycling "Brassica rapa" (Wisconsin Fast Plants)

    Science.gov (United States)

    Kelly, Martin G.; Terrana, Sebastian

    2004-01-01

    In this paper, we demonstrate that rapid cycling "Brassica rapa" (Wisconsin Fast Plants) can be used in inquiry-based, student ecological fieldwork. We are the first to describe age-specific survival for field-grown Fast Plants and identify life history traits associated with individual survival. This experiment can be adapted by educators as a…

  17. Rapid transfer of photosynthetic carbon through the plant-soil system in differently managed species-rich grasslands

    NARCIS (Netherlands)

    Deyn, de G.B.; Quirk, H.; Oakley, S.; Ostle, N.J.; Bartgett, R.D.

    2011-01-01

    Plant-soil interactions are central to short-term carbon (C) cycling through the rapid transfer of recently assimilated C from plant roots to soil biota. In grassland ecosystems, changes in C cycling are likely to be influenced by land use and management that changes vegetation and the associated

  18. Fast Plants for Finer Science--An Introduction to the Biology of Rapid-Cycling Brassica Campestris (rapa) L.

    Science.gov (United States)

    Tomkins, Stephen P.; Williams, Paul H.

    1990-01-01

    Rapid-cycling brassicas can be used in the classroom to teach concepts such as plant growth, tropisms, floral reproduction, pollination, embryonic development, and plant genetics. Directions on how to obtain them for classroom use and how they may be grown are included. Practical physiology and genetics exercises are listed. (KR)

  19. Indirect rapid prototyping of sol-gel hybrid glass scaffolds for bone regeneration - Effects of organic crosslinker valence, content and molecular weight on mechanical properties.

    Science.gov (United States)

    Hendrikx, Stephan; Kascholke, Christian; Flath, Tobias; Schumann, Dirk; Gressenbuch, Mathias; Schulze, F Peter; Hacker, Michael C; Schulz-Siegmund, Michaela

    2016-04-15

    We present a series of organic/inorganic hybrid sol-gel derived glasses, made from a tetraethoxysilane-derived silica sol (100% SiO2) and oligovalent organic crosslinkers functionalized with 3-isocyanatopropyltriethoxysilane. The material was susceptible to heat sterilization. The hybrids were processed into pore-interconnected scaffolds by an indirect rapid prototyping method, described here for the first time for sol-gel glass materials. A large panel of polyethylene oxide-derived 2- to 4-armed crosslinkers of molecular weights ranging between 170 and 8000Da were incorporated and their effect on scaffold mechanical properties was investigated. By multiple linear regression, 'organic content' and the 'content of ethylene oxide units in the hybrid' were identified as the main factors that determined compressive strength and modulus, respectively. In general, 3- and 4-armed crosslinkers performed better than linear molecules. Compression tests and cell culture experiments with osteoblast-like SaOS-2 cells showed that macroporous scaffolds can be produced with compressive strengths of up to 33±2MPa and with a pore structure that allows cells to grow deep into the scaffolds and form mineral deposits. Compressive moduli between 27±7MPa and 568±98MPa were obtained depending on the hybrid composition and problems associated with the inherent brittleness of sol-gel glass materials could be overcome. SaOS-2 cells showed cytocompatibility on hybrid glass scaffolds and mineral accumulation started as early as day 7. On day 14, we also found mineral accumulation on control hybrid glass scaffolds without cells, indicating a positive effect of the hybrid glass on mineral accumulation. We produced a hybrid sol-gel glass material with significantly improved mechanical properties towards an application in bone regeneration and processed the material into macroporous scaffolds of controlled architecture by indirect rapid prototyping. We were able to produce macroporous materials

  20. Regenerable Photovoltaic Devices with a Hydrogel-Embedded Microvascular Network

    Science.gov (United States)

    Koo, Hyung-Jun; Velev, Orlin D.

    2013-01-01

    Light-driven degradation of photoactive molecules could be one of the major obstacles to stable long term operation of organic dye-based solar light harvesting devices. One solution to this problem may be mimicking the regeneration functionality of a plant leaf. We report an organic dye photovoltaic system that has been endowed with such microfluidic regeneration functionality. A hydrogel medium with embedded channels allows rapid and uniform supply of photoactive reagents by a convection-diffusion mechanism. A washing-activation cycle enables reliable replacement of the organic component in a dye-sensitized photovoltaic system. Repetitive restoration of photovoltaic performance after intensive device degradation is demonstrated. PMID:23912814

  1. A modified MS2 bacteriophage plaque reduction assay for the rapid screening of antiviral plant extracts.

    Science.gov (United States)

    Cock, Ian; Kalt, F R

    2010-07-01

    Traditional methods of screening plant extracts and purified components for antiviral activity require up to a week to perform, prompting the need to develop more rapid quantitative methods to measure the ability of plant based preparations to block viral replication. We describe an adaption of an MS2 plaque reduction assay for use in S. aureus. MS2 bacteriophage was capable of infecting and replicating in B. cereus, S. aureus and F + E. coli but not F- E. coli. Indeed, both B. cereus and S. aureus were more sensitive to MS2 induced lysis than F+ E. coli. When MS2 bacteriophage was mixed with Camellia sinensis extract (1 mg/ml), Scaevola spinescens extract (1 mg/ml) or Aloe barbadensis juice and the mixtures inoculated into S. aureus, the formation of plaques was reduced to 8.9 ± 3.8%, 5.4 ± 2.4% and 72.7 ± 20.9% of the untreated MS2 control values respectively. The ability of the MS2 plaque reduction assay to detect antiviral activity in these known antiviral plant preparations indicates its suitability as an antiviral screening tool. An advantage of this assay compared with traditionally used cytopathic effect reduction assays and replicon based assays is the more rapid acquisition of results. Antiviral activity was detected within 24 h of the start of testing. The MS2 assay is also inexpensive and non-pathogenic to humans making it ideal for initial screening studies or as a simulant for pathogenic viruses.

  2. An efficient in vitro shoot regeneration from immature inflorescence and ex vitro rooting of Arnebia hispidissima (Lehm). DC. - A red dye (Alkannin) yielding plant.

    Science.gov (United States)

    Phulwaria, Mahendra; Shekhawat, N S

    2013-07-01

    Arnebia hispidissima, which belongs to the family Boraginaceae, is an important medicinal and dye yielding plant. The alkannin, a red dye, are root-specific secondary metabolites of A. hispidissima. Shoots were regenerated from callus derived from immature inflorescence explants obtained from field grown plants. MS medium containing 4.52 μM 2, 4-D and 3.33 μM BAP was found to be most effective for the proliferation of callus, induced on medium containing 4.52 μM 2, 4-D. Maximum number (43.1 ± 0.25) with average length (5.2 ± 0.23) of shoots regenerated when callus was transferred to MS medium supplemented with 1.11 μM BAP, 1.16 μM Kin and 0.57 μM IAA. About 75.5 % of in vitro regenerated shoots were rooted on half-strength MS medium supplemented with 9.84 μM of IBA and 200 mg l(-1) of activated charcoal. In comparison to in vitro, higher percent (90.2 %) of shoots were rooted under ex vitro conditions when treated with IBA (0.98 mM) for 5 min. Plantlets rooted in vitro as well as ex vitro were acclimatized successfully under the green house conditions. Ex vitro rooted plants exhibited higher survival percentage (75 %) as compared to in vitro rooted plantlets (60 %). Present study may be applicable in the large-scale root-specific red dye (alkannin) production via root induction under ex vitro condition.

  3. Multicellular genesis of leaf primordium was demonstrated via chimaeric transgenic plant of maize (Zea mays L.) regenerated from Type II calli.

    Science.gov (United States)

    Xu, Zi-Qin; Huang, Xuan; Feng, Chao; Tian, Na; Xu, Dan; Feng, Shu-Zhen

    2010-10-01

    Type-II embryonic calli were induced from immature embryos of maize (Zea mays L.) genotype YD and bombarded with beta-glucuronidase gene. Bombarded calli were proliferated on normal N6 medium for 2 weeks at 26°C in the dark and selected on N6 medium containing 1 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 5 mg/l phosphinothricin (PPT) but without casamino acids and proline under the same conditions for 14 days. Regeneration was carried out on hormone-free MS medium containing 5 mg/l phosphinothricin at 26°C under 3000 lux illumination. Plants over 8 cm were transplanted into soil and sprayed with 250 mg/l phosphinothricin when two new leaves appeared. Except normal transgenic plants, chimaeric transgenics also were regenerated in the present work. The expression pattern of beta-glucuronidase gene in leaves of chimaeric transgenic plant revealed that more than one cell formed leaf primordium at the initial stage, and filial cells stemed from each cell in leaf primordium arranged in a row longitudinally from leaf base to leaf apex. There was a clear boundary as a straight line between the area formed by transformed cells and the area formed by normal cells. A hypothesis was put forward that the primitive cells in leaf primordium divided in a longitudinal style, resulted in leaf elongation, then the filial cells divided transversally and synchronously toward the outside to broaden the leaf.

  4. Realizing the potential of rapid-cycling Brassica as a model system for use in plant biology research

    Science.gov (United States)

    Musgrave, M. E.

    2000-01-01

    Rapid-cycling Brassica populations were initially developed as a model for probing the genetic basis of plant disease. Paul Williams and co-workers selected accessions of the six main species for short time to flower and rapid seed maturation. Over multiple generations of breeding and selection, rapid-cycling populations of each of the six species were developed. Because of their close relationship with economically important Brassica species, rapid-cycling Brassica populations, especially those of B. rapa (RCBr) and B. oleracea, have seen wide application in plant and crop physiology investigations. Adding to the popularity of these small, short-lived plants for research applications is their extensive use in K-12 education and outreach.

  5. Regeneration of Pelargonium in vitro

    Directory of Open Access Journals (Sweden)

    Agnieszka Wojtania

    2013-12-01

    Full Text Available Pelargonium sp. has been a subject of numerous studies to deterimine the effec tiveness of in vitro techniques to produce a large number of pathogen-free plants. Regeneration of pelargonium plants from the different initial explants as well via organogenesis as via somatic embryogenesis has been obtained. The most effective adventitious shoot formation has been achieved from shoot tips and axillary buds using cytokinin or cytokinin/auxin combinations. Leaf explants, whose general have lower organogenic potency, regenerate better in the presence of thidiazuron. This growth regulator stimulate the somatic embryos production from hypocotyl and cotyledone explants too. The main problem in tissue culture propagation of Pelargonium has been the high tendency to formation of vigorously growing callus with low organogenic potency and rapid senescence of cultures. Moreover, the significant differen ces in requirements to the medium composition (minerals, organic compounds and growth regulators between Pelargonium cultivars has been observed. This makes difficult to develop an universaI method of Pelargonium micropropagation.

  6. Limb regeneration.

    Science.gov (United States)

    Simon, András; Tanaka, Elly M

    2013-01-01

    Limb regeneration is observed in certain members of the animal phyla. Some animals keep this ability during their entire life while others lose it at some time during development. How do animals regenerate limbs? Is it possible to find unifying, conserved mechanisms of limb regeneration or have different species evolved distinct means of replacing a lost limb? How is limb regeneration similar or different to limb development? Studies on many organisms, including echinoderms, arthropods, and chordates have provided significant knowledge about limb regeneration. In this focus article, we concentrate on tetrapod limb regeneration as studied in three model amphibians: newts, axolotls, and frogs. We review recent progress on tissue interactions during limb regeneration, and place those findings into an evolutionary context. Copyright © 2012 Wiley Periodicals, Inc.

  7. A Rapid and Reliable Method for Total Protein Extraction from Succulent Plants for Proteomic Analysis.

    Science.gov (United States)

    Lledías, Fernando; Hernández, Felipe; Rivas, Viridiana; García-Mendoza, Abisaí; Cassab, Gladys I; Nieto-Sotelo, Jorge

    2017-08-01

    Crassulacean acid metabolism plants have some morphological features, such as succulent and reduced leaves, thick cuticles, and sunken stomata that help them prevent excessive water loss and irradiation. As molecular constituents of these morphological adaptations to xeric environments, succulent plants produce a set of specific compounds such as complex polysaccharides, pigments, waxes, and terpenoids, to name a few, in addition to uncharacterized proteases. Since all these compounds interfere with the analysis of proteins by electrophoretic techniques, preparation of high quality samples from these sources represents a real challenge. The absence of adequate protocols for protein extraction has restrained the study of this class of plants at the molecular level. Here, we present a rapid and reliable protocol that could be accomplished in 1 h and applied to a broad range of plants with reproducible results. We were able to obtain well-resolved SDS/PAGE protein patterns in extracts from different members of the subfamilies Agavoideae (Agave, Yucca, Manfreda, and Furcraea), Nolinoideae (Dasylirion and Beucarnea), and the Cactaceae family. This method is based on the differential solubility of contaminants and proteins in the presence of acetone and pH-altered solutions. We speculate about the role of saponins and high molecular weight carbohydrates to produce electrophoretic-compatible samples. A modification of the basic protocol allowed the analysis of samples by bidimensional electrophoresis (2DE) for proteomic analysis. Furostanol glycoside 26-O-β-glucosidase (an enzyme involved in steroid saponin synthesis) was successfully identified by mass spectrometry analysis and de novo sequencing of a 2DE spot from an Agave attenuata sample.

  8. Plant regeneration through somatic embryogenesis in root-derived callus of ginseng (Panax ginseng C. A. Meyer).

    Science.gov (United States)

    Chang, W C; Hsing, Y I

    1980-05-01

    Callus culture was initiated from expiants of mature root tissues of ginseng (Panax ginseng C.A. Meyer) on MS medium enriched with 2,4-D. The ageing callus produced numerous embryoids in this medium. Reculture of these embryoids in media (1/2 MS or B5) supplemented with benzyladenine and gibberellic acid resulted in profuse plantlet regeneration.

  9. Effect of low temperature on dormancy breaking and growth after planting in lily bulblets regenerated in vitro

    NARCIS (Netherlands)

    Langens-Gerrits, M.M.; Miller, W.B.; Croes, A.; Klerk, de G.J.M.

    2003-01-01

    Lilies regenerating on scale segments may develop dormancy in vitro depending on the culture conditions. The dormancy is broken by storage for several weeks at a low temperature ( 5 degreesC). The effect of the low temperature on sprouting, time of leaf emergence and further bulb growth was studied.

  10. Influence of Removal of a Non-native Tree Species Mimosa caesalpiniifolia Benth. on the Regenerating Plant Communities in a Tropical Semideciduous Forest Under Restoration in Brazil

    Science.gov (United States)

    Podadera, Diego S.; Engel, Vera L.; Parrotta, John A.; Machado, Deivid L.; Sato, Luciane M.; Durigan, Giselda

    2015-11-01

    Exotic species are used to trigger facilitation in restoration plantings, but this positive effect may not be permanent and these species may have negative effects later on. Since such species can provide a marketable product (firewood), their harvest may represent an advantageous strategy to achieve both ecological and economic benefits. In this study, we looked at the effect of removal of a non-native tree species ( Mimosa caesalpiniifolia) on the understory of a semideciduous forest undergoing restoration. We assessed two 14-year-old plantation systems (modified "taungya" agroforestry system; and mixed plantation using commercial timber and firewood tree species) established at two sites with contrasting soil properties in São Paulo state, Brazil. The experimental design included randomized blocks with split plots. The natural regeneration of woody species (height ≥0.2 m) was compared between managed (all M. caesalpiniifolia trees removed) and unmanaged plots during the first year after the intervention. The removal of M. caesalpiniifolia increased species diversity but decreased stand basal area. Nevertheless, the basal area loss was recovered after 1 year. The management treatment affected tree species regeneration differently between species groups. The results of this study suggest that removal of M. caesalpiniifolia benefited the understory and possibly accelerated the succession process. Further monitoring studies are needed to evaluate the longer term effects on stand structure and composition. The lack of negative effects of tree removal on the natural regeneration indicates that such interventions can be recommended, especially considering the expectations of economic revenues from tree harvesting in restoration plantings.

  11. [Study on tissue culture and plant regeneration of the stem-tips and buds of Periploca forrestii].

    Science.gov (United States)

    Liu, Zhi-Fei; Gao, Jie; Niu, Ya-Hui; Shi, Lei; Chen, Tao-Tao

    2011-11-01

    To establish the rapid propagation systems of the stem-tips and buds of Periploca forrestii. Inserted the stem-tips and buds of Periploca forrestii into MS medium with different concentrations of 6-BA, NAA and 2.4-D and induced them growing into complete plants. The optimal culture medium for bud induction is MS + 6-BA 1.0 mg/L + NAA 0.3 mg/L and the bud induction rate can reach 86.29%. The optimal culture medium for stem-tips induction is MS +6-BA 2.0 mg/L + NAA 0.5 mg/L and the bud induction rate can reach 86.29%. The optimal culture medium for bud multiplication is MS + 6-BA 2.0 mg/L + NAA 0.1 mg/L and the multiplication coefficient can reach 2.10. The best rooting medium is 1/2 MS + IBA 0.5 mg/L and the rooting rate is 53.33%.

  12. Molecular characterization and expression analysis of SERK1 and SERK2 in Brassica napus L.: implication for microspore embryogenesis and plant regeneration.

    Science.gov (United States)

    Ahmadi, Behzad; Masoomi-Aladizgeh, Farhad; Shariatpanahi, Mehran E; Azadi, Pejman; Keshavarz-Alizadeh, Mehdi

    2016-01-01

    The BnSERK1 and BnSERK2 are involved in the process of microspore embryogenesis induction, development, and plantlet regeneration. Little is known about regulatory role of somatic embryogenesis-related kinase (SERK) genes family in the induction of microspore embryogenesis, development and plant regeneration. In this study, the expression of two SERK genes (SERK1 and SERK2) was assessed during the microspore embryogenesis and plantlet regeneration in Brassica napus L. The BnSERK1 was severely up-regulated 1-5 days following microspore culture and its expression drastically decreased in the globular-heart and also torpedo staged microspore-derived embryos (MDEs). In addition, high levels of BnSERK1 transcript were detected in the MDE maturation phase and in the roots and shoots of the regenerated plantlets which indicates a broader role(s) of BnSERK1 in the organ formation, rather than being specific to the embryogenesis. Results of partial sequencing indicated that the BnSERK1 shares a conserved serine-threonine kinase catalytic domain and exhibited 95 % similarity with AtSERK1, CsSERK1, BrSERK1, NaSERK1, and NbSERK1. A steady increase in the expression of BnSERK2 was observed during the MDE initiation and development so that, the highest expression was noted in the MDE maturation phase i.e., late cotyledonary MDEs. Our results also indicated low amounts of BnSERK2 transcript at the onset of rhyzogenesis but significantly higher expression in the developing roots. In contrast, the BnSERK2 strongly up-regulated during the both initially and developed shoots. The BnSERK2 shares highly conserved LRR-RLK domain when compared with different species tested so that, high homology (100 %) was noticed with BrSERK2. Based on our findings, MDE formation and plantlet regeneration seem to be correlated with both BnSERK1 and BnSERK2 expression.

  13. A novel method for rapid and non-invasive detection of plants senescence using delayed fluorescence technique

    Science.gov (United States)

    Zhang, Lingrui; Xing, Da; Wang, Junsheng; Zeng, Lizhang; Li, Qiang

    2007-05-01

    Plants senescence is a phase of plants ontogeny marked by declining photosynthetic activity that is paralleled by a decline in chloroplast function. The photosystem II ( PSII ) in a plant is considered the primary site where light-induced delayed fluorescence (DF) is produced. With the leaves of Catharanthus roseus (Catharanthus roseus (L.) G.Don) as testing models, we have studied the effects of plants senescence induced by dark and/or exogenous hormones treatments on characteristics of DF by using a home-made portable DF detection system, which can enable various DF parameters, such as DF decay kinetic curve and DF intensity, to be rapidly produced for the plants in a short time. The results show that the changes in DF intensity of green plants can truly reflect the changes in photosynthetic capacity and chlorophyll content. Therefore, DF may be used an important means of evaluating in vivo plants senescence physiology. The changes in DF intensity may provide a new approach for the rapid and early detection of plants senescence caused by age or other senescence-related factors. DF technique could be potential useful for high throughput screening and less time-consuming and automated identifying the interesting mutants with genetic modifications that change plants senescence progress.

  14. Salts and nutrients present in regenerated waters induce changes in water relations, antioxidative metabolism, ion accumulation and restricted ion uptake in Myrtus communis L. plants.

    Science.gov (United States)

    Acosta-Motos, José R; Alvarez, Sara; Barba-Espín, Gregorio; Hernández, José A; Sánchez-Blanco, María J

    2014-12-01

    The use of reclaimed water (RW) constitutes a valuable strategy for the efficient management of water and nutrients in landscaping. However, RW may contain levels of toxic ions, affecting plant production or quality, a very important aspect for ornamental plants. The present paper evaluates the effect of different quality RWs on physiological and biochemical parameters and the recovery capacity in Myrtus communis L. plants. M. communis plants were submitted to 3 irrigation treatments with RW from different sources (22 weeks): RW1 (1.7 dS m(-1)), RW2 (4.0 dS m(-1)) and RW3 (8.0 dS m(-1)) and one control (C, 0.8 dS m(-1)). During a recovery period of 11 weeks, all plants were irrigated with the control water. The RW treatments did not negatively affect plant growth, while RW2 even led to an increase in biomass. After recovery,only plants irrigated with RW3 showed some negative effects on growth, which was related to a decrease in the net photosynthesis rate, higher Na accumulation and a reduction in K levels. An increase in salinity was accompanied by decreases in leaf water potential, relative water content and gas exchange parameters, and increases in Na and Cl uptake. Plants accumulated Na in roots and restricted its translocation to the aerial part. The highest salinity levels produced oxidative stress, as seen from the rise in electrolyte leakage and lipid peroxidation. The use of regenerated water together with carefully managed drainage practices, which avoid the accumulation of salt by the substrate, will provide economic and environmental benefits.

  15. Rapid genetic transformation of sweetpotato ( Ipomoea batatas (L ...

    African Journals Online (AJOL)

    It indicated an auxin to cytokinin treatment could improve the regeneration of transgenic calluses. This rapid organogenesis-based transformation strategy represents an important improvement over existing methods and will facilitate producing large-scale transgenic sweetpotato plants the genetic improvement of a crop ...

  16. Rapid demise and recovery of plant ecosystems across the end-Permian extinction event

    Science.gov (United States)

    Hochuli, Peter A.; Hermann, Elke; Vigran, Jorunn Os; Bucher, Hugo; Weissert, Helmut

    2010-12-01

    The end-Permian extinction event was the most pronounced biotic and ecological crisis in the history of the Earth. It is assumed that over 80% of marine genera disappeared, and that this event had a major impact on the evolution of marine organisms. The impact of this event on terrestrial biota is poorly known and a matter of controversial discussions. In contrast to the fundamental changes in marine fauna most major groups of plants range from the Late Palaeozoic into the Mesozoic. Consequently the impact of the end-Permian extinction event on the evolution of plants was often regarded as minor. However, major changes in the composition of the plant communities have been documented and a number of catastrophic scenarios have been envisioned — including the almost total destruction of plant ecosystems. Based on expanded sections from the Southern Barents Sea (Northern Norway) we trace mid-latitudinal terrestrial ecosystems across the Permo-Triassic transition with a time resolution in the order of 10 kyr, based on a high resolution C org-isotope stratigraphy. Our results show that the floral turnovers are linked with major changes in the C-isotope record and hence with global carbon cycling. The palynological records document the successive steps in the evolution of terrestrial ecosystems. After gradual changes during the latest Permian, plant ecosystems suffered from a major environmental perturbation leading to a rapid turnover from gymnosperm dominated ecosystems to assemblages dominated by lycopods. The dominance of the lycopods, expressed in a spore-spike, represents a relatively short-lived event in the order of 10 kyr. This perturbation of the terrestrial ecosystems preceded the globally recognized negative δ 13C org isotope spike by up to 100 kyr. It coincides with a first end-Permian negative shift of the C-isotope curve and was probably induced by a first major perturbation of the chemistry of the atmosphere, related to the onset of the volcanic

  17. Combining Near-Infrared Spectroscopy and Chemometrics for Rapid Recognition of an Hg-Contaminated Plant

    Directory of Open Access Journals (Sweden)

    Bang-Cheng Tang

    2016-01-01

    Full Text Available The feasibility of rapid recognition of an Hg-contaminated plant as a soil pollution indicator was investigated using near-infrared spectroscopy (NIRS and chemometrics. The stem and leave of a native plant, Miscanthus floridulus (Labill. Warb. (MFLW, were collected from Hg-contaminated areas (n1=125 as well as from regular areas (n2=116. The samples were dried and crushed and the powders were sieved through an 80-mesh sieve. Reference analysis of Hg levels was performed using inductively coupled plasma-atomic emission spectrometry (ICP-AES. The actual Hg contents of contaminated and normal samples were 16.2–30.5 and 0.0–0.1 mg/Kg, respectively. The NIRS measurements of impacted sample powders were collected in the mode of reflectance. The DUPLEX algorithm was utilized to split the NIRS data into representative training and test sets. Different spectral preprocessing methods were performed to remove the unwanted and noncomposition-correlated spectral variations. Classification models were developed using partial least squares discrimination analysis (PLSDA based on the raw, smoothed, second-order derivative (D2, and standard normal variate (SNV data, respectively. The prediction accuracy obtained by PLSDA with each data preprocessing option was 100%, indicating pattern recognition of Hg-contaminated MFLW samples using NIRS data was in perfect consistence with the ICP-AES results. NIRS combined with chemometrics will provide a tool to screen the Hg-contaminated MFLW, which can be potentially used as an indicator of soil pollution.

  18. Plant regeneration of Korean wild ginseng (Panax ginseng Meyer) mutant lines induced by γ-irradiation (60Co) of adventitious roots

    Science.gov (United States)

    Zhang, Jun-Ying; Sun, Hyeon-Jin; Song, In-Ja; Bae, Tae-Woong; Kang, Hong-Gyu; Ko, Suk-Min; Kwon, Yong-Ik; Kim, Il-Woung; Lee, Jaechun; Park, Shin-Young; Lim, Pyung-Ok; Kim, Yong Hwan; Lee, Hyo-Yeon

    2014-01-01

    An efficient in vitro protocol has been established for somatic embryogenesis and plantlet conversion of Korean wild ginseng (Panax ginseng Meyer). Wild-type and mutant adventitious roots derived from the ginseng produced calluses on Murashige and Skoog (MS) medium supplemented with 0.5 mg/L 2,4-dichlorophenoxyacetic acid and 0.3 mg/L kinetin; 53.3% of the explants formed callus. Embryogenic callus proliferation and somatic embryo induction occurred on MS medium containing 0.5 mg/L 2,4-dichlorophenoxyacetic acid. The induced somatic embryos further developed to maturity on MS medium with 5 mg/L gibberellic acid, and 85% of them germinated. The germinated embryos were developed to shoots and elongated on MS medium with 5 mg/L gibberellic acid. The shoots developed into plants with well-developed taproots on one-third strength Schenk and Hildebrandt basal medium supplemented with 0.25 mg/L 1-naphthaleneacetic acid. When the plants were transferred to soil, about 30% of the regenerated plants developed into normal plants. PMID:25378998

  19. Plant regeneration of Korean wild ginseng (Panax ginseng Meyer) mutant lines induced by γ-irradiation ((60)Co) of adventitious roots.

    Science.gov (United States)

    Zhang, Jun-Ying; Sun, Hyeon-Jin; Song, In-Ja; Bae, Tae-Woong; Kang, Hong-Gyu; Ko, Suk-Min; Kwon, Yong-Ik; Kim, Il-Woung; Lee, Jaechun; Park, Shin-Young; Lim, Pyung-Ok; Kim, Yong Hwan; Lee, Hyo-Yeon

    2014-07-01

    An efficient in vitro protocol has been established for somatic embryogenesis and plantlet conversion of Korean wild ginseng (Panax ginseng Meyer). Wild-type and mutant adventitious roots derived from the ginseng produced calluses on Murashige and Skoog (MS) medium supplemented with 0.5 mg/L 2,4-dichlorophenoxyacetic acid and 0.3 mg/L kinetin; 53.3% of the explants formed callus. Embryogenic callus proliferation and somatic embryo induction occurred on MS medium containing 0.5 mg/L 2,4-dichlorophenoxyacetic acid. The induced somatic embryos further developed to maturity on MS medium with 5 mg/L gibberellic acid, and 85% of them germinated. The germinated embryos were developed to shoots and elongated on MS medium with 5 mg/L gibberellic acid. The shoots developed into plants with well-developed taproots on one-third strength Schenk and Hildebrandt basal medium supplemented with 0.25 mg/L 1-naphthaleneacetic acid. When the plants were transferred to soil, about 30% of the regenerated plants developed into normal plants.

  20. Plant regeneration of Korean wild ginseng (Panax ginseng Meyer mutant lines induced by γ-irradiation (60Co of adventitious roots

    Directory of Open Access Journals (Sweden)

    Jun-Ying Zhang

    2014-07-01

    Full Text Available An efficient in vitro protocol has been established for somatic embryogenesis and plantlet conversion of Korean wild ginseng (Panax ginseng Meyer. Wild-type and mutant adventitious roots derived from the ginseng produced calluses on Murashige and Skoog (MS medium supplemented with 0.5 mg/L 2,4-dichlorophenoxyacetic acid and 0.3 mg/L kinetin; 53.3% of the explants formed callus. Embryogenic callus proliferation and somatic embryo induction occurred on MS medium containing 0.5 mg/L 2,4-dichlorophenoxyacetic acid. The induced somatic embryos further developed to maturity on MS medium with 5 mg/L gibberellic acid, and 85% of them germinated. The germinated embryos were developed to shoots and elongated on MS medium with 5 mg/L gibberellic acid. The shoots developed into plants with well-developed taproots on one-third strength Schenk and Hildebrandt basal medium supplemented with 0.25 mg/L 1-naphthaleneacetic acid. When the plants were transferred to soil, about 30% of the regenerated plants developed into normal plants.

  1. An efficient in vitro shoot regeneration from leaf petiolar explants and ex vitro rooting of Bixa orellana L.- A dye yielding plant.

    Science.gov (United States)

    Mohammed, Arifullah; Chiruvella, Kishore K; Namsa, Nima D; Ghanta, Rama Gopal

    2015-07-01

    Bixa orellana L. (Bixaceae) is a multipurpose tree grown for the production of commercially important dyes. In the present study, an efficient, reproducible protocol was developed for direct plant regeneration from in vitro derived petiole explants of Bixa orellana L. Murashige and Skoog medium (MS) supplemented with 2-isopentenyl adenine (9.8 μM) and naphthalene acetic acid (10.7 μM) was found to be optimum for production of high frequency of shoot organogenesis. Subculturing of the shoots onto the fresh MS medium containing similar concentrations of 2-iP (9.8 μM) and NAA (10.7 μM) produced elongated shoots. Elongated shoots when placed onto MS medium supplemented with 1.7 μM indole-3-acetic acid and 14.7 μM 2-iP produced optimal rooting. Rooted plantlets were acclimatized and transplanted to the field successfully. Histological investigation revealed the origin of shoot primordia, from sub-epidermal cells of petiole explants. The regeneration protocol developed in this study can be useful for mass in vitro propagation and effective genetic transformation of commercially important edible dye yielding tree species.

  2. Development of a Rapid and Simple Method to Remove Polyphenols from Plant Extracts

    Directory of Open Access Journals (Sweden)

    Imali Ranatunge

    2017-01-01

    Full Text Available Polyphenols are secondary metabolites of plants, which are responsible for prevention of many diseases. Polyvinylpolypyrrolidone (PVPP has a high affinity towards polyphenols. This method involves the use of PVPP column to remove polyphenols under centrifugal force. Standards of gallic acid, epigallocatechin gallate, vanillin, and tea extracts (Camellia sinensis were used in this study. PVPP powder was packed in a syringe with different quantities. The test samples were layered over the PVPP column and subjected to centrifugation. Supernatant was tested for the total phenol content. The presence of phenolic compounds and caffeine was screened by HPLC and measuring the absorbance at 280. The antioxidant capacity of standards and tea extracts was compared with the polyphenol removed fractions using DPPH scavenging assay. No polyphenols were found in polyphenolic standards or tea extracts after PVPP treatment. The method described in the present study to remove polyphenols is simple, inexpensive, rapid, and efficient and can be employed to investigate the contribution of polyphenols present in natural products to their biological activity.

  3. Plant Growth and Development: An Outline for a Unit Structured Around the Life Cycle of Rapid-Cycling Brassica Rapa.

    Science.gov (United States)

    Becker, Wayne M.

    This outline is intended for use in a unit of 10-12 lectures on plant growth and development at the introductory undergraduate level as part of a course on organismal biology. The series of lecture outlines is structured around the life cycle of rapid-cycling Brassica rapa (RCBr). The unit begins with three introductory lectures on general plant…

  4. Rapid transfer of photosynthetic carbon through the plant-soil system in differently managed species-rich grasslands

    Directory of Open Access Journals (Sweden)

    G. B. De Deyn

    2011-05-01

    Full Text Available Plant-soil interactions are central to short-term carbon (C cycling through the rapid transfer of recently assimilated C from plant roots to soil biota. In grassland ecosystems, changes in C cycling are likely to be influenced by land use and management that changes vegetation and the associated soil microbial communities. Here we tested whether changes in grassland vegetation composition resulting from management for plant diversity influences short-term rates of C assimilation and transfer from plants to soil microbes. To do this, we used an in situ 13C-CO2 pulse-labelling approach to measure differential C uptake among different plant species and the transfer of the plant-derived 13C to key groups of soil microbiota across selected treatments of a long-term plant diversity grassland restoration experiment. Results showed that plant taxa differed markedly in the rate of 13C assimilation and concentration: uptake was greatest and 13C concentration declined fastest in Ranunculus repens, and assimilation was least and 13C signature remained longest in mosses. Incorporation of recent plant-derived 13C was maximal in all microbial phosopholipid fatty acid (PLFA markers at 24 h after labelling. The greatest incorporation of 13C was in the PLFA 16:1ω5, a marker for arbuscular mycorrhizal fungi (AMF, while after 1 week most 13C was retained in the PLFA18:2ω6,9 which is indicative of assimilation of plant-derived 13C by saprophytic fungi. Our results of 13C assimilation and transfer within plant species and soil microbes were consistent across management treatments. Overall, our findings suggest that plant diversity restoration management may not directly affect the C assimilation or retention of C by individual plant taxa or groups of soil microbes, it can impact on the fate of recent C by changing their relative abundances

  5. Rapid and sensitive hormonal profiling of complex plant samples by liquid chromatography coupled to electrospray ionization tandem mass spectrometry

    Directory of Open Access Journals (Sweden)

    Müller Maren

    2011-11-01

    Full Text Available Abstract Background Plant hormones play a pivotal role in several physiological processes during a plant's life cycle, from germination to senescence, and the determination of endogenous concentrations of hormones is essential to elucidate the role of a particular hormone in any physiological process. Availability of a sensitive and rapid method to quantify multiple classes of hormones simultaneously will greatly facilitate the investigation of signaling networks in controlling specific developmental pathways and physiological responses. Due to the presence of hormones at very low concentrations in plant tissues (10-9 M to 10-6 M and their different chemistries, the development of a high-throughput and comprehensive method for the determination of hormones is challenging. Results The present work reports a rapid, specific and sensitive method using ultrahigh-performance liquid chromatography coupled to electrospray ionization tandem spectrometry (UPLC/ESI-MS/MS to analyze quantitatively the major hormones found in plant tissues within six minutes, including auxins, cytokinins, gibberellins, abscisic acid, 1-amino-cyclopropane-1-carboxyic acid (the ethylene precursor, jasmonic acid and salicylic acid. Sample preparation, extraction procedures and UPLC-MS/MS conditions were optimized for the determination of all plant hormones and are summarized in a schematic extraction diagram for the analysis of small amounts of plant material without time-consuming additional steps such as purification, sample drying or re-suspension. Conclusions This new method is applicable to the analysis of dynamic changes in endogenous concentrations of hormones to study plant developmental processes or plant responses to biotic and abiotic stresses in complex tissues. An example is shown in which a hormone profiling is obtained from leaves of plants exposed to salt stress in the aromatic plant, Rosmarinus officinalis.

  6. Plant regeneration from cotyledonary explants of Eucalyptus camaldulensis Regeneração de plantas de Eucalyptus camaldulensis a partir das explantes cotiledonares

    Directory of Open Access Journals (Sweden)

    Roberson Dibax

    2005-08-01

    Full Text Available Breeding methods based on genetic transformation techniques need to be implemented for Eucalyptus camaldulensis to shorten the long breeding cycles and avoid manipulation of adult trees; that requires the development of plant regeneration protocols enabling development of plants from transformed tissues. The present work aimed to optimise the regeneration process already established for the species. Cotyledonary leaves of E. camaldulensis were cultured in MS medium supplemented with naphthaleneacetic acid (NAA and 6-benzylaminopurine (BAP combinations. The most efficient treatment for bud indirect regeneration (2.7 µmol L-1 NAA and 4.44 µmol L-1 BAP was used for further experiments. When explants were kept in the dark during the first 30 days, the percentage of explants forming calluses increased and explant necrosis was reduced in comparison with light-cultured explants. Mineral medium modifications were compared and half-strength MS mineral medium turned out to be as efficient as full-strength medium, producing 54% and 47% of explants with buds, respectively. For shoot elongation, MS medium with half-strength nitrate and ammonium salts, and 0.2% activated charcoal yielded rooted shoots 1 to 8 cm high after one month. The procedure is an efficient protocol for E. camadulensis plant regeneration, reducing the stages necessary for the obtention of complete plants.A implementação, para espécies florestais, de técnicas de melhoramento baseadas em métodos de transformação genética, permitirá reduzir os longos ciclos de melhoramento e evitar a manipulação de árvores adultas. Isto implica dispor de um protocolo de regeneração que permita o desenvolvimento de plantas a partir de tecidos transformados. Este trabalho teve como objetivo otimizar este protocolo de regeneração para Eucalyptus camaldulensis. Folhas cotiledonares foram cultivadas em meio de cultura MS suplementado com combinações de ácido naftalenoacético (ANA e 6

  7. Plant regeneration by somatic embryogenesis from cultured immature embryos of oak (Querem robur L.) and linden (Tilia cordata Mill.).

    Science.gov (United States)

    Chalupa, V

    1990-11-01

    Embryogenic cultures and somatic embryos were obtained from immature zygotic embryos of oak (Quercus robur L.) cultured on a modified MS medium and WPM containing BAP (1 mg·l(-1)) and GA3 (1 mg·l(-1)) or BAP and IBA. Germination and conversion of oak somatic embryos into plantlets was achieved on WPM containing a reduced concentration of cytokinin. Linden (Tilia cordata Mill.) somatic embryos developed in embryogenic tissues initiated from immature zygotic embryos cultured on a modified MS medium supplemented with 2,4-D (0.3-2.0 mg·l(-1)). Germination of linden somatic embryos and plantlet formation occurred on MS medium containing a low concentration of IBA. Oak and linden plantlets produced from somatic embryos were successfully established in soil. Somatic embryos and plantlets were also regenerated from embryogenic cultures of Quercus petraea and Tilia platyphyllos.

  8. Assessing the Potential of Low-Cost 3D Cameras for the Rapid Measurement of Plant Woody Structure

    Directory of Open Access Journals (Sweden)

    Charles Nock

    2013-11-01

    Full Text Available Detailed 3D plant architectural data have numerous applications in plant science, but many existing approaches for 3D data collection are time-consuming and/or require costly equipment. Recently, there has been rapid growth in the availability of low-cost, 3D cameras and related open source software applications. 3D cameras may provide measurements of key components of plant architecture such as stem diameters and lengths, however, few tests of 3D cameras for the measurement of plant architecture have been conducted. Here, we measured Salix branch segments ranging from 2–13 mm in diameter with an Asus Xtion camera to quantify the limits and accuracy of branch diameter measurement with a 3D camera. By scanning at a variety of distances we also quantified the effect of scanning distance. In addition, we also test the sensitivity of the program KinFu for continuous 3D object scanning and modeling as well as other similar software to accurately record stem diameters and capture plant form (<3 m in height. Given its ability to accurately capture the diameter of branches >6 mm, Asus Xtion may provide a novel method for the collection of 3D data on the branching architecture of woody plants. Improvements in camera measurement accuracy and available software are likely to further improve the utility of 3D cameras for plant sciences in the future.

  9. Adventitious shoot regeneration from in vitro stem explants of ...

    African Journals Online (AJOL)

    An efficient in vitro plant regeneration system from stem explants was established in Phellodendron amurense. Factors influencing shoot regeneration from stems including culture medium type, combinations of plant growth regulators and carbon source in the medium were investigated. Adventitious shoot regeneration was ...

  10. Solasodine accumulation in regenerated plants of Solanum torvum Sw Acúmulo de solasodina em plantas micropropagadas de Solanum torvum Sw

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    C.B Moreira

    2010-03-01

    Full Text Available A nodal segment culture was developed in order to assess Solanum torvum Sw. regeneration and solasodine levels. The influence of auxins (indoleacetic acid, 1-Naphthaleneacetic acid and benzyl adenine on S. torvum growth in micropropagation was investigated. A nodal segment culture was initiated with seeds germinated in MS basal medium added of GA3 and grown in different concentrations of IAA, IAA + BAP and NAA + BAP. Sixty-day-old plants from the in vitro culture were collected, frozen and lyophilized; then, the methyl orange method was used to quantify solasodine for the spectrophotometric assay. The best results regarding plant regeneration and solasodine accumulation were obtained by using the MS basal medium without addition of plant growth regulators; however, there was great production of calluses presenting friable bases. Based on these results, cell cultures can be initiated from such calluses with application of other auxins and cytokinins to enhance solasodine production, besides different elicitors, light intensities and sucrose concentrations.A regeneração de Solanum torvum e a avaliação do conteúdo de solasodina foram os objetivos de cultura de segmentos nodais. A influência de auxinas (ácido 3-indolacético, ácido naftalenoacético e de 6-benzilaminopurina no crescimento de S. torvum na micropropagação foi investigado. Cultura de segmentos nodais foi iniciada por sementes germinadas em meio básico MS acrescido de GA3 e cultivadas em diferentes concentrações de AIA, AIA + BAP e ANA + BAP. Plantas da cultura in vitro com 60 dias foram coletadas, congeladas e liofilizadas e o método de alaranjado de metila foi utilizado para quantificação de solasodina para o ensaio espectrofotométrico. Os melhores resultados para regeneração vegetal e acúmulo de solasodina foram alcançados no meio MS sem adição de reguladores de crescimento havendo, porém grande produção de calos de base friáveis. Esses resultados mostram

  11. Esterase polymorphism and the analysis of genetic diversity and structure in cactus populations descended from Cereus peruvianus plants regenerated in vitro.

    Science.gov (United States)

    Sala, Juliana; Mangolin, Claudete Aparecida; Franzoni, Juliana; Machado, Maria de Fátima Pires da Silva

    2011-04-01

    The genetic structure of Cereus peruvianus populations descended from cultivated plants (F(1) populations) and from plants regenerated in vitro (R(1) populations) was analyzed using α- and β-esterase isozymes in native PAGE. The estimated proportion of polymorphic loci was higher (50%) in the R(1) populations than the F(1) populations (42.85%). The mean observed (0.5599) and expected (0.5620) heterozygosity in R(1) descendents was also higher than the rates in F(1) descendents (H (o) = 0.4142; H (e) = 0.4977). A low level of population differentiation was detected in R(1) descendents (F (st) = 0.05). In contrast, population differentiation was high in F(1) descendents (0.2583). Esterase analysis using PAGE showed that artificial selection by silvicultural management provides high genetic diversity and a large genetic basis for C. peruvianus, whereas in vitro selection from callus tissue culture involves an increase of heterozygosity levels in descendents from somaclones and a low level of interpopulational divergence.

  12. New protocol for the fast induction of adventicious buds and plants regeneration of banana cv. ‘Grande Naine’ (Musa AAA

    Directory of Open Access Journals (Sweden)

    Lourdes R. García

    2006-01-01

    Full Text Available A protocol for the quick formation of adventitious buds in banana cv. ‘Grande Naine’ and the development of plants was developed. Several concentrations of 6-Benzylaminopurine (6-BAP and 1-phenyl-3-(-1,2,3-thidiazol-5-ylurea (TDZ were studied in the culture media for the induction of these structures, evaluating some explants sizes for multiplication. The highest number of adventitious buds was obtained when TDZ was used in the culture medium, with only two subcultures needed to develop explants with adventitious buds. The explants of 1mm3 were selected for the multiplication and a medium of MS salts with 1.0 mg.l-1 thiamine HCl and 2.0% sucrose for plant regeneration, reaching the values of 96%. The use of this protocol could be a very useful alternative in genetic improvement programs with the cv. ´Grande naine` using the mutation induction and somaclonal variation. Key words: organogenesis, thidiazuron, tissue culture

  13. Rapid plant identification using species- and group-specific primers targeting chloroplast DNA.

    Directory of Open Access Journals (Sweden)

    Corinna Wallinger

    Full Text Available Plant identification is challenging when no morphologically assignable parts are available. There is a lack of broadly applicable methods for identifying plants in this situation, for example when roots grow in mixture and for decayed or semi-digested plant material. These difficulties have also impeded the progress made in ecological disciplines such as soil- and trophic ecology. Here, a PCR-based approach is presented which allows identifying a variety of plant taxa commonly occurring in Central European agricultural land. Based on the trnT-F cpDNA region, PCR assays were developed to identify two plant families (Poaceae and Apiaceae, the genera Trifolium and Plantago, and nine plant species: Achillea millefolium, Fagopyrum esculentum, Lolium perenne, Lupinus angustifolius, Phaseolus coccineus, Sinapis alba, Taraxacum officinale, Triticum aestivum, and Zea mays. These assays allowed identification of plants based on size-specific amplicons ranging from 116 bp to 381 bp. Their specificity and sensitivity was consistently high, enabling the detection of small amounts of plant DNA, for example, in decaying plant material and in the intestine or faeces of herbivores. To increase the efficacy of identifying plant species from large number of samples, specific primers were combined in multiplex PCRs, allowing screening for multiple species within a single reaction. The molecular assays outlined here will be applicable manifold, such as for root- and leaf litter identification, botanical trace evidence, and the analysis of herbivory.

  14. [Effect of the extremely weak alternating magnetic fields on the regeneration of planarians and the gravitropic response of plants].

    Science.gov (United States)

    Belova, N A; Ermakov, A M; Znobishcheva, A V; Skrebnitskaia, L K; Lednev, V V

    2010-01-01

    The influence of extremely weak alternating magnetic fields (EW AMF) directed colinearly to the static Earth magnetic field on the rate of regeneration of planarians and the rate of gravitropic response in the stem segments of flax has been studied. The value of bioeffects of EW AMF is determined by the parameter gamma B(AC)/f, where y is the gyromagnetic ratio of the magnetic moments induced by the orbital movements of electrons in atoms, and B(AC) and f correspond to magnetic induction and frequency of the alternating magnetic component. It was shown that the magnitude of bioeffects depends on the amplitude (at fixed 1000 Hz - frequency) and frequency (at fixed 192 nT - amplitude) of the alternating component. Maxima of bioeffects are observed at gamma B(AC)/f = 0.9; 2.75, and minor maxima gamma B(AC)/f = 4.5; 6.1. The bioeffects are absent at gamma B(AC)/f = 1.8, 3.8, 5.3, 6.7. The positions of the maxima and minima of bioeffects correspond to the theoretical prediction (at gamma = 14000 Hz/microT). Primary targets for the EW AMF of this type are the magnetic moments induced by the orbital movements of electrons in atoms.

  15. Rapid identification of a narcotic plant Papaver bracteatum using flow cytometry

    OpenAIRE

    Aragane, Masako; Watanabe, Daisuke; Nakajima, Jun’ichi; YOSHIDA, Masao; Yoshizawa, Masao; Abe, Tomohiro; Nishiyama, Rei; Suzuki, Jin; Moriyasu, Takako; Nakae, Dai; Sudo, Hiroshi; Sato, Hiroyuki; Hishida, Atuyuki; Kawahara, Nobuo; Makabe, So

    2014-01-01

    In May 2011, numerous poppy plants closely resembling Papaver bracteatum Lindl., a type of narcotic plant that is illegal in Japan, were distributed directly from several large flower shops or through online shopping throughout Japan, including the Tokyo Metropolitan area. In order to better identify the narcotic plants, the relative nuclear DNA content at the vegetative stage was measured by flow cytometric (FCM) analysis in 3 closely-related species of the genus Papaver section Oxytona, nam...

  16. Plant Lesions Promote the Rapid Multiplication of Escherichia coli O157:H7 on Postharvest Lettuce▿

    OpenAIRE

    Brandl, M. T.

    2008-01-01

    Several outbreaks of Escherichia coli O157:H7 infections have been associated with minimally processed leafy vegetables in the United States. Harvesting and processing cause plant tissue damage. In order to assess the role of plant tissue damage in the contamination of leafy greens with E. coli O157:H7, the effect of mechanical, physiological, and plant disease-induced lesions on the growth of this pathogen on postharvest romaine lettuce was investigated. Within only 4 h after inoculation, th...

  17. A simple, rapid, and reliable protocol to localize hydrogen peroxide in large plant organs by DAB-mediated tissue printing.

    Science.gov (United States)

    Liu, Yong-Hua; Offler, Christina E; Ruan, Yong-Ling

    2014-01-01

    Hydrogen peroxide (H2O2) is a major reactive oxygen species (ROS) and plays diverse roles in plant development and stress responses. However, its localization in large and thick plant organs (e.g., stem, roots, and fruits), other than leaves, has proven to be challenging due to the difficulties for the commonly used H2O2-specific chemicals, such as 3,3'-diaminobenzidine (DAB), cerium chloride (CeCl3), and 2',7'-dichlorofluorescin diacetate (H2DCF-DA), to penetrate those organs. Theoretically, the reaction of endogenous H2O2 with these chemicals could be facilitated by using thin organ sections. However, the rapid production of wound-induced H2O2 associated with this procedure inevitably disturbs the original distribution of H2O2 in vivo. Here, by employing tomato seedling stems and fruits as testing materials, we report a novel, simple, and rapid protocol to localize H2O2 in those organs using DAB-mediated tissue printing. The rapidity of the protocol (within 15 s) completely avoided the interference of wound-induced H2O2 during experimentation. Moreover, the H2O2 signal on the printing was stable for at least 1 h with no or little background produced. We conclude that DAB-mediated tissue printing developed here provide a new feasible and reliable method to localize H2O2 in large plant organs, hence should have broad applications in studying ROS biology.

  18. Rapid high-level production of functional HIV broadly neutralizing monoclonal antibodies in transient plant expression systems.

    Directory of Open Access Journals (Sweden)

    Yvonne Rosenberg

    Full Text Available Passive immunotherapy using anti-HIV broadly neutralizing monoclonal antibodies (mAbs has shown promise as an HIV treatment, reducing mother-to-child-transmission (MTCT of simian/human immunodeficiency virus (SHIV in non-human primates and decreasing viral rebound in patients who ceased receiving anti-viral drugs. In addition, a cocktail of potent mAbs may be useful as mucosal microbicides and provide an effective therapy for post-exposure prophylaxis. However, even highly neutralizing HIV mAbs used today may lose their effectiveness if resistance occurs, requiring the rapid production of new or engineered mAbs on an ongoing basis in order to counteract the viral resistance or the spread of a certain HIV-1 clade in a particular region or patient. Plant-based expression systems are fast, inexpensive and scalable and are becoming increasingly popular for the production of proteins and monoclonal antibodies. In the present study, Agrobacterium-mediated transient transfection of plants, utilizing two species of Nicotiana, have been tested to rapidly produce high levels of an HIV 89.6PΔ140env and several well-studied anti-HIV neutralizing monoclonal antibodies (b12, 2G12, 2F5, 4E10, m43, VRC01 or a single chain antibody construct (m9, for evaluation in cell-based viral inhibition assays. The protein-A purified plant-derived antibodies were intact, efficiently bound HIV envelope, and were equivalent to, or in one case better than, their counterparts produced in mammalian CHO or HEK-293 cells in both neutralization and antibody dependent viral inhibition assays. These data indicate that transient plant-based transient expression systems are very adaptable and could rapidly generate high levels of newly identified functional recombinant HIV neutralizing antibodies when required. In addition, they warrant detailed cost-benefit analysis of prolonged incubation in plants to further increase mAb production.

  19. Rapid high-level production of functional HIV broadly neutralizing monoclonal antibodies in transient plant expression systems.

    Science.gov (United States)

    Rosenberg, Yvonne; Sack, Markus; Montefiori, David; Forthal, Donald; Mao, Lingjun; Hernandez-Abanto, Segundo; Urban, Lori; Landucci, Gary; Fischer, Rainer; Jiang, Xiaoming

    2013-01-01

    Passive immunotherapy using anti-HIV broadly neutralizing monoclonal antibodies (mAbs) has shown promise as an HIV treatment, reducing mother-to-child-transmission (MTCT) of simian/human immunodeficiency virus (SHIV) in non-human primates and decreasing viral rebound in patients who ceased receiving anti-viral drugs. In addition, a cocktail of potent mAbs may be useful as mucosal microbicides and provide an effective therapy for post-exposure prophylaxis. However, even highly neutralizing HIV mAbs used today may lose their effectiveness if resistance occurs, requiring the rapid production of new or engineered mAbs on an ongoing basis in order to counteract the viral resistance or the spread of a certain HIV-1 clade in a particular region or patient. Plant-based expression systems are fast, inexpensive and scalable and are becoming increasingly popular for the production of proteins and monoclonal antibodies. In the present study, Agrobacterium-mediated transient transfection of plants, utilizing two species of Nicotiana, have been tested to rapidly produce high levels of an HIV 89.6PΔ140env and several well-studied anti-HIV neutralizing monoclonal antibodies (b12, 2G12, 2F5, 4E10, m43, VRC01) or a single chain antibody construct (m9), for evaluation in cell-based viral inhibition assays. The protein-A purified plant-derived antibodies were intact, efficiently bound HIV envelope, and were equivalent to, or in one case better than, their counterparts produced in mammalian CHO or HEK-293 cells in both neutralization and antibody dependent viral inhibition assays. These data indicate that transient plant-based transient expression systems are very adaptable and could rapidly generate high levels of newly identified functional recombinant HIV neutralizing antibodies when required. In addition, they warrant detailed cost-benefit analysis of prolonged incubation in plants to further increase mAb production.

  20. In vitro regeneration of sweet potato (Ipomoea batatas (L.) Lam ...

    African Journals Online (AJOL)

    Ipomoea batatas (L.) Lam.) yield. The establishment of in vitro regeneration protocol for this plant is essential to improve it through tissue culture and genetic engineering. The objective of this study was to establish in vitro regeneration protocol ...

  1. [Rapid diagnostics of early phosphorus deficiency in mini-cucumber plants under protected cultivation by near infrared spectroscopy].

    Science.gov (United States)

    Shi, Ji-yong; Zou, Xiao-bo; Zhao, Jie-wen; Mao, Han-ping; Wang, Kai-liang; Chen, Zheng-wei; Huang, Xiao-wei

    2011-12-01

    The morphological symptom of phosphorus deficiency at early stage is similar to the appearance of leaf aging process in preliminary phase, so that visual diagnostics of phosphorus deficiency in mini-cucumber plants at early stage is practically impossible. Near infrared reflectance spectra contain information about differences in compositions of leaf tissues between phosphorus-deficient plants and healthy plants. In the present paper, near infrared reflectance spectroscopy was used to provide diagnostic information on phosphorus deficiency of mini-cucumber plants grown under non-soil conditions. Near infrared spectra was collected from 90 leaves of mini-cucumber plants. Raw cucumber spectra was preprocessed by SNV and divided into 27 intervals. The top 10 principal components (PCs) were extracted as the input of BP-ANN classifiers by principal component analysis (PCA) while the values of nutrient deficient were used as the output variables of BP-ANN and three layers BP-ANN discrimination model was built. The best experiment results were based on the top 3 principal components of No. 7 interval when the spectra was divided into 27 intervals and identification rates of the ANN model are 100% in both training set and the prediction set. The overall results show that NIR spectroscopy combined with BP-ANN can be efficiently utilized for rapid and early diagnostics of phosphorus deficiency in mini-cucumber plants.

  2. Rapid transcriptional plasticity of duplicated gene clusters enables a clonally reproducing aphid to colonise diverse plant species.

    Science.gov (United States)

    Mathers, Thomas C; Chen, Yazhou; Kaithakottil, Gemy; Legeai, Fabrice; Mugford, Sam T; Baa-Puyoulet, Patrice; Bretaudeau, Anthony; Clavijo, Bernardo; Colella, Stefano; Collin, Olivier; Dalmay, Tamas; Derrien, Thomas; Feng, Honglin; Gabaldón, Toni; Jordan, Anna; Julca, Irene; Kettles, Graeme J; Kowitwanich, Krissana; Lavenier, Dominique; Lenzi, Paolo; Lopez-Gomollon, Sara; Loska, Damian; Mapleson, Daniel; Maumus, Florian; Moxon, Simon; Price, Daniel R G; Sugio, Akiko; van Munster, Manuella; Uzest, Marilyne; Waite, Darren; Jander, Georg; Tagu, Denis; Wilson, Alex C C; van Oosterhout, Cock; Swarbreck, David; Hogenhout, Saskia A

    2017-02-13

    The prevailing paradigm of host-parasite evolution is that arms races lead to increasing specialisation via genetic adaptation. Insect herbivores are no exception and the majority have evolved to colonise a small number of closely related host species. Remarkably, the green peach aphid, Myzus persicae, colonises plant species across 40 families and single M. persicae clonal lineages can colonise distantly related plants. This remarkable ability makes M. persicae a highly destructive pest of many important crop species. To investigate the exceptional phenotypic plasticity of M. persicae, we sequenced the M. persicae genome and assessed how one clonal lineage responds to host plant species of different families. We show that genetically identical individuals are able to colonise distantly related host species through the differential regulation of genes belonging to aphid-expanded gene families. Multigene clusters collectively upregulate in single aphids within two days upon host switch. Furthermore, we demonstrate the functional significance of this rapid transcriptional change using RNA interference (RNAi)-mediated knock-down of genes belonging to the cathepsin B gene family. Knock-down of cathepsin B genes reduced aphid fitness, but only on the host that induced upregulation of these genes. Previous research has focused on the role of genetic adaptation of parasites to their hosts. Here we show that the generalist aphid pest M. persicae is able to colonise diverse host plant species in the absence of genetic specialisation. This is achieved through rapid transcriptional plasticity of genes that have duplicated during aphid evolution.

  3. Pneumatic hydrodynamics influence transplastomic protein yields and biological responses duringin vitroshoot regeneration ofNicotiana tabacumcallus: Implications for bioprocess routes to plant-made biopharmaceuticals.

    Science.gov (United States)

    Barretto, Sherwin S; Michoux, Franck; Hellgardt, Klaus; Nixon, Peter J

    2017-01-15

    Transplastomic plants are capable of high-yield production of recombinant biopharmaceutical proteins. Plant tissue culture combines advantages of agricultural cultivation with the bioprocess consistency associated with suspension culture. Overexpression of recombinant proteins through regeneration of transplastomic Nicotiana tabacum shoots from callus tissue in RITA ® temporary immersion bioreactors has been previously demonstrated. In this study we investigated the hydrodynamics of periodic pneumatic suspension of liquid medium during temporary immersion culture (4 min aeration every 8 h), and the impact on biological responses and transplastomic expression of fragment C of tetanus toxin (TetC). Biomass was grown under a range of aeration rates for 3, 20 and 40-day durations. Growth, mitochondrial activity (a viability indicator) and TetC protein yields were correlated against the hydrodynamic parameters, shear rate and energy dissipation rate (per kg of medium). A critical aeration rate of 440 ml min -1 was identified, corresponding to a shear rate of 96.7 s -1 , pneumatic power input of 8.8 mW kg -1 and initial 20-day pneumatic energy dissipation of 127 J kg -1 , at which significant reductions in biomass accumulation and mitochondrial activity were observed. There was an exponential decline in TetC yields with increasing aeration rates at 40 days, across the entire range of conditions tested. These observations have important implications for the optimisation and scale-up of transplastomic plant tissue culture bioprocesses for biopharmaceutical production.

  4. Rapid Py-GC/MS assessment of the structural alterations of lignins in genetically modified plants

    NARCIS (Netherlands)

    Rencoret, Jorge; Del Río, José Carlos; Nierop, Klaas G J; Gutiérrez, Ana; Ralph, John

    Genetic modifications for perturbing the lignin pathway in three different species of angiosperm plants, including non-woody (Arabidopsis and alfalfa) and woody (poplar) plants, were readily evaluated by analytical pyrolysis coupled to gas chromatography-mass spectrometry (Py-GC/MS). Pyrolysis

  5. Estimation of plant-available nitrogen in soils using rapid chemical and biological methods

    NARCIS (Netherlands)

    Velthof, G.L.; Oenema, O.

    2010-01-01

    The relationships between potential laboratory indices for plant-available nitrogen (N) and the plant N uptake in a pot experiment with ryegrass were assessed for 13 mineral soils and 2 peat soils. The methods included aerobic soil incubation, soil incubation in a bioreactor, hot potassium chloride

  6. Rapid identification of a narcotic plant Papaver bracteatum using flow cytometry.

    Science.gov (United States)

    Aragane, Masako; Watanabe, Daisuke; Nakajima, Jun'ichi; Yoshida, Masao; Yoshizawa, Masao; Abe, Tomohiro; Nishiyama, Rei; Suzuki, Jin; Moriyasu, Takako; Nakae, Dai; Sudo, Hiroshi; Sato, Hiroyuki; Hishida, Atuyuki; Kawahara, Nobuo; Makabe, So; Nakamura, Ikuo; Mii, Masahiro

    2014-10-01

    In May 2011, numerous poppy plants closely resembling Papaver bracteatum Lindl., a type of narcotic plant that is illegal in Japan, were distributed directly from several large flower shops or through online shopping throughout Japan, including the Tokyo Metropolitan area. In order to better identify the narcotic plants, the relative nuclear DNA content at the vegetative stage was measured by flow cytometric (FCM) analysis in 3 closely-related species of the genus Papaver section Oxytona, namely P. orientale, P. pseudo-orientale, and P. bracteatum, based on the difference between the chromosome numbers of these species. The results showed that the nuclear DNA content differed between these 3 species, and that most of the commercially distributed plants examined in this study could be identified as P. bracteatum. The remaining plants were P. pseudo-orientale, a non-narcotic plant. In addition, the FCM results for the identification of P. bracteatum completely agreed with the results obtained by the morphological analysis, the inter-genic spacer sequence of rpl16-rpl14 (PS-ID sequence) of chloroplast DNA, and the presence of thebaine. These results clearly indicate the usefulness of FCM analysis for the identification of P. bracteatum plants, including when they are in their vegetative stage.

  7. The relative importance of rapid evolution for plant-microbe interactions depends on ecological context.

    Science.gov (United States)

    Terhorst, Casey P; Lennon, Jay T; Lau, Jennifer A

    2014-06-22

    Evolution can occur on ecological time-scales, affecting community and ecosystem processes. However, the importance of evolutionary change relative to ecological processes remains largely unknown. Here, we analyse data from a long-term experiment in which we allowed plant populations to evolve for three generations in dry or wet soils and used a reciprocal transplant to compare the ecological effect of drought and the effect of plant evolutionary responses to drought on soil microbial communities and nutrient availability. Plants that evolved under drought tended to support higher bacterial and fungal richness, and increased fungal : bacterial ratios in the soil. Overall, the magnitudes of ecological and evolutionary effects on microbial communities were similar; however, the strength and direction of these effects depended on the context in which they were measured. For example, plants that evolved in dry environments increased bacterial abundance in dry contemporary environments, but decreased bacterial abundance in wet contemporary environments. Our results suggest that interactions between recent evolutionary history and ecological context affect both the direction and magnitude of plant effects on soil microbes. Consequently, an eco-evolutionary perspective is required to fully understand plant-microbe interactions.

  8. Rapid plant-cover establishment on gold mine tailings in southern New Zealand: glasshouse screening trials.

    Science.gov (United States)

    Schroeder, K; Rufaut, C G; Smith, C; Mains, D; Craw, D

    2005-01-01

    The use of a short-term vegetation cover to temporarily control the negative environmental effects of inactive tailings ponds is notfrequently practiced during operational mining, but could have some merit This article reports on a glasshouse trial designed to examine some of the issues associated with short-term vegetation: fast germination of a high proportion of seed, the ability of seedlings to survive in unamended substrates, and potentially toxic substrate. Five nonindigenous plant species were tested--barley (Hordeum vulgare), rye corn (Secale cereale), Italian ryegrass (Lolium multifiorum), red clover (Trifolium pratense), and lucerne (Medicago sativa)--in five different types of substrate: unamended tailings, tailings and fertilizer, tailings and greenwaste, biosolid-blend compost, and local topsoil. The nutrient and heavy metal status (As, Cu, Cd, Ni, Pb) of each substrate type was determined Plant species performance was monitored over 14 wk Substrate metal concentrations were low except for As, which was elevated in all substrate types. Plants in unamended tailings grew less vigorously than plants in tailings and compost or in topsoil. Plant performance in tailings and fertiliser was greatly suppressed following a high fertilization rate. Metal uptake in plants was highest for As (0.4-77 mg kg(-1) DW) and Cu (5.3-50.3 mg kg(-1) DW). Future field trials are necessary to authenticate findings, but barley and rye corn are promising species for a short-term tailings cover.

  9. Rapid, high-yield production in plants of individualized idiotype vaccines for non-Hodgkin's lymphoma.

    Science.gov (United States)

    Bendandi, M; Marillonnet, S; Kandzia, R; Thieme, F; Nickstadt, A; Herz, S; Fröde, R; Inogés, S; Lòpez-Dìaz de Cerio, A; Soria, E; Villanueva, H; Vancanneyt, G; McCormick, A; Tusé, D; Lenz, J; Butler-Ransohoff, J-E; Klimyuk, V; Gleba, Y

    2010-12-01

    Animal and clinical studies with plant-produced single-chain variable fragment lymphoma vaccines have demonstrated specific immunogenicity and safety. However, the expression levels of such fragments were highly variable and required complex engineering of the linkers. Moreover, the downstream processing could not be built around standard methods like protein A affinity capture. We report a novel vaccine manufacturing process, magnifection, devoid of the above-mentioned shortcomings and allowing consistent and efficient expression in plants of whole immunoglobulins (Igs). Full idiotype (Id)-containing IgG molecules of 20 lymphoma patients and 2 mouse lymphoma models were expressed at levels between 0.5 and 4.8 g/kg of leaf biomass. Protein A affinity capture purification yielded antigens of pharmaceutical purity. Several patient Igs produced in plants showed specific cross-reactivity with sera derived from the same patients immunized with hybridoma-produced Id vaccine. Mice vaccinated with plant- or hybridoma-produced Igs showed comparable protection levels in tumor challenge studies. This manufacturing process is reliable and robust, the manufacturing time from biopsy to vaccine is antibodies in plants, providing 50- to 1000-fold higher yields than alternative plant expression methods.

  10. Rapid modulation of ultraviolet shielding in plants is influenced by solar ultraviolet radiation and linked to alterations in flavonoids.

    Science.gov (United States)

    Barnes, Paul W; Tobler, Mark A; Keefover-Ring, Ken; Flint, Stephan D; Barkley, Anne E; Ryel, Ronald J; Lindroth, Richard L

    2016-01-01

    The accumulation of ultraviolet (UV)-absorbing compounds (flavonoids and related phenylpropanoids) and the resultant decrease in epidermal UV transmittance (TUV ) are primary protective mechanisms employed by plants against potentially damaging solar UV radiation and are critical components of the overall acclimation response of plants to changing solar UV environments. Whether plants can adjust this UV sunscreen protection in response to rapid changes in UV, as occurs on a diurnal basis, is largely unexplored. Here, we use a combination of approaches to demonstrate that plants can modulate their UV-screening properties within minutes to hours, and these changes are driven, in part, by UV radiation. For the cultivated species Abelmoschus esculentus, large (30-50%) and reversible changes in TUV occurred on a diurnal basis, and these adjustments were associated with changes in the concentrations of whole-leaf UV-absorbing compounds and several quercetin glycosides. Similar results were found for two other species (Vicia faba and Solanum lycopersicum), but no such changes were detected in Zea mays. These findings reveal a much more dynamic UV-protection mechanism than previously recognized, raise important questions concerning the costs and benefits of UV-protection strategies in plants and have practical implications for employing UV to enhance crop vigor and quality in controlled environments. © 2015 John Wiley & Sons Ltd.

  11. The effect of growth regulators (BAP and IBA on regeneration, proliferation and rooting of Natanz pears plant using in vitro technique

    Directory of Open Access Journals (Sweden)

    Abbas Safarnejad

    2016-09-01

    Full Text Available Pear (Pyrus sp. is an important fruit growing in temperate and cold areas that due to suave and high economic value were cultured since ancient in Iran. The purpose of this study was to identify the best media for fast propagation of healthy plants of Pyrus sp. using tissue culture technique. At first, the pears samples were collected from the garden of Astan Quds Razavi and lateral and terminal buds were used as explants. For sterilization 0.1% mercuric chloride for 3 minutes, 70% ethanol for 1 minute was the best treatment. A Complete Randomized Design experiment was carried out with 3 replications. Results showed that Regeneration initiated on MSbasal medium supplemented with 3 mg/l BAP + 0.01 mg/l IBA. The most multiplication was on MS medium supplemented with 0.1 mg/l IBA+ 3 mg/l BAP. The highest shoot length was observed in MS medium supplemented with 0.01 mg/l IBA+ 3 mg/l BAP. The results of ANOVA showed no significant differences between rooting treatment. The rooted plantlets transferred to Jf pot for adaptation and then transferred to soil.

  12. Gas purge-microsyringe extraction: a rapid and exhaustive direct microextraction technique of polycyclic aromatic hydrocarbons from plants.

    Science.gov (United States)

    Wang, Juan; Yang, Cui; Li, Huijie; Piao, Xiangfan; Li, Donghao

    2013-12-17

    Gas purge-microsyringe extraction (GP-MSE) is a rapid and exhaustive microextraction technique for volatile and semivolatile compounds. In this study, a theoretical system of GP-MSE was established by directly extracting and analyzing 16 kinds of polycyclic aromatic hydrocarbons (PAHs) from plant samples. On the basis of theoretical consideration, a full factorial experimental design was first used to evaluate the main effects and interactions of the experimental parameters affecting the extraction efficiency. Further experiments were carried out to determine the extraction kinetics and desorption temperature-dependent. The results indicated that three factors, namely desorption temperature (temperature of sample phase) Td, extraction time t, and gas flow rate u, had a significantly positive effect on the extraction efficiency of GP-MSE for PAHs. Extraction processes of PAHs in plant samples followed by first-order kinetics (relative coefficient R(2) of simulation curves were 0.731-1.000, with an average of 0.958 and 4.06% relative standard deviation), and obviously depended on the desorption temperature. Furthermore, the effect of the matrix was determined from the difference in Eapp,d. Finally, satisfactory recoveries of 16 PAHs were obtained using optimal parameters. The study demonstrated that GP-MSE could provide a rapid and exhaustive means of direct extraction of PAHs from plant samples. The extraction kinetics were similar that of the inverse process of the desorption kinetics of the sample phase. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. Intercellular communication in plants: evidence for two rapidly transmitted systemic signals generated in response to electromagnetic field stimulation in tomato.

    Science.gov (United States)

    Beaubois, Elisabeth; Girard, Sebastien; Lallechere, Sebastien; Davies, Eric; Paladian, Françoise; Bonnet, Pierre; Ledoigt, Gerard; Vian, Alain

    2007-07-01

    Exposing all of a wild-type tomato plant to electromagnetic radiation evoked rapid and substantial accumulation of basic leucine-zipper transcription factor (bZIP) mRNA in the terminal leaf (#4) with kinetics very similar to that seen in response to wounding, while in the abscisic acid (ABA) mutant (Sitiens), the response was more rapid, but transient. Submitting just the oldest leaf (#1) of a wild-type plant to irradiation evoked bZIP mRNA accumulation both locally in the exposed leaf and systemically in the unexposed (distant) leaf #4, although systemic accumulation was delayed somewhat. Accumulation of Pin2 mRNA was less than bZIP in both the exposed and distant leaves in wild type, but there was no delay in the systemic response. In Sitiens, bZIP mRNA accumulation was far less than in wild type in both local and distant leaves, while Pin2 mRNA accumulation was stronger in the exposed leaf, but totally prevented in the systemic leaf. In the jasmonic acid (JA) mutant (JL-5) and in wild-type plants treated with the ABA biosynthesis inhibitor, naproxen, responses were similar to those in the ABA mutant, while treatment of the exposed leaf with calcium antagonists totally abolished both local and systemic increases in bZIP transcript accumulation.

  14. [A hydroponic cultivation system for rapid high-yield transient protein expression in Nicotiana plants under laboratory conditions].

    Science.gov (United States)

    Mo, Qianzhen; Mai, Rongjia; Yang, Zhixiao; Chen, Minfang; Yang, Tiezhao; Lai, Huafang; Yang, Peiliang; Chen, Qiang; Zhou, Xiaohong

    2012-06-01

    To develop a hydroponic Nicotiana cultivation system for rapid and high-yield transient expression of recombinant proteins under laboratory conditions. To establish the hydroponic cultivation system, several parameters were examined to define the optimal conditions for the expression of recombinant proteins in plants. We used the green fluorescent protein (GFP) and the geminiviral plant transient expression vector as the model protein/expression vector. We examined the impact of Nicotiana species, the density and time of Agrobacterium infiltration, and the post-infiltration growth period on the accumulation of GFP. The expression levels of GFP in Nicotiana leaves were then examined by Western blotting and ELISA. Our data indicated that a hydroponic Nicotiana cultivation system with a light intensity of 9000 LX/layer, a light cycle of 16 h day/8 h night, a temperature regime of 28 degrees celsius; day/21 degrees celsius; night, and a relative humidity of 80% could support the optimal plant growth and protein expression. After agroinfiltration with pBYGFPDsRed.R/LBA4404, high levels of GFP expression were observed in both N. benthamiana and N. tobaccum (cv. Yuyan No.5) plants cultured with this hydroponic cultivation system. An optimal GFP expression was achieved in both Nicotiana species leaves 4 days after infiltration by Agrobacterium with an OD(600) of 0.8. At a given time point, the average biomass of N. tobaccum (cv. Yuyan No.5) was significantly higher than that of N. benthamiana. The leaves from 6-week-old N. benthamiana plants and 5-week-old N. tobaccum (cv. Yuyan No.5) plants could be the optimal material for agroinfiltration. We have established a hydroponic cultivation system that allows robust growth of N. benthamiana and N. tobaccum (cv. Yuyan No.5) plants and the optimal GFP expression in the artificial climate box.

  15. High-frequency regeneration via multiple shoot induction of an elite recalcitrant cotton (Gossypium hirsutum L. cv Narashima) by using embryo apex

    Science.gov (United States)

    Pathi, Krishna Mohan; Tuteja, Narendra

    2013-01-01

    Cotton (Gossypium hirsutum L.) is one of the most commercially important fiber crops in the world. Compared with other crops, cotton represents a recalcitrant species for regeneration protocols. The development of efficient and rapid regeneration protocol for elite Indian cotton variety could help improve the quality characteristics and biotic or abiotic stress tolerance. Here we report a novel regeneration protocol in Indian cotton cultivar Narashima. The maximum number of multiple shoots obtained was 16 per explants, performance which has never been achieved in any prior reports. The embryo apex explants were isolated from 2 d old in vitro growing seedlings. Explants were cultured on MS medium containing different plant growth regulator combinations in order to induce multiple shoots. Among the tested combinations, the 2 mg/l of 6-benzylaminopurine (BAP) and 2 mg/l kinetin (KIN) proved to be most suited for achieving the maximum number of multiple shoots. The elongation of multiple shoots was obtained in media supplemented with gibberellic acid (GA3). The regenerated plants were successfully hardened in earthen pots after adequate acclimatization. This method avoids callus tissue, the stage of regeneration which may lead to somaclonal variation. The important feature of the presented method is shortening of regeneration time, as well as the induction of a high number of multiple shoots per explants. The present protocol may provide an efficient and rapid regeneration tool for obtaining more stable transformants from embryo apex explants of Indian cotton cultivar Narashima. PMID:23221745

  16. Low Cost, High Capacity Regenerable Sorbent for Carbon Dioxide Capture from Existing Coal-fired Power Plants

    Energy Technology Data Exchange (ETDEWEB)

    Alptekin, Gokhan [TDA Research, Inc., Wheat Ridge, CO (United States); Jayaraman, Ambalavanan [TDA Research, Inc., Wheat Ridge, CO (United States); Dietz, Steven [TDA Research, Inc., Wheat Ridge, CO (United States)

    2016-03-03

    In this project TDA Research, Inc (TDA) has developed a new post combustion carbon capture technology based on a vacuum swing adsorption system that uses a steam purge and demonstrated its technical feasibility and economic viability in laboratory-scale tests and tests in actual coal derived flue gas. TDA uses an advanced physical adsorbent to selectively remove CO2 from the flue gas. The sorbent exhibits a much higher affinity for CO2 than N2, H2O or O2, enabling effective CO2 separation from the flue gas. We also carried out a detailed process design and analysis of the new system as part of both sub-critical and super-critical pulverized coal fired power plants. The new technology uses a low cost, high capacity adsorbent that selectively removes CO2 in the presence of moisture at the flue gas temperature without a need for significant cooling of the flue gas or moisture removal. The sorbent is based on a TDA proprietary mesoporous carbon that consists of surface functionalized groups that remove CO2 via physical adsorption. The high surface area and favorable porosity of the sorbent also provides a unique platform to introduce additional functionality, such as active groups to remove trace metals (e.g., Hg, As). In collaboration with the Advanced Power and Energy Program of the University of California, Irvine (UCI), TDA developed system simulation models using Aspen PlusTM simulation software to assess the economic viability of TDA’s VSA-based post-combustion carbon capture technology. The levelized cost of electricity including the TS&M costs for CO2 is calculated as $116.71/MWh and $113.76/MWh for TDA system integrated with sub-critical and super-critical pulverized coal fired power plants; much lower than the $153.03/MWhand $147.44/MWh calculated for the corresponding amine based systems. The cost of CO2 captured for TDA’s VSA based system is $38

  17. A simple, rapid and reliable protocol to localize hydrogen peroxide in large plant organs by DAB-mediated tissue printing

    Directory of Open Access Journals (Sweden)

    Yonghua eLiu

    2014-12-01

    Full Text Available Hydrogen peroxide (H2O2 is a major reactive oxygen species (ROS and plays diverse roles in plant development and stress responses. However, its localization in large and thick plant organs (e.g. stem, roots and fruits, other than leaves, has proven to be challenging due to the difficulties for the commonly used H2O2-specific chemicals, such as 3, 3’-diaminobenzidine (DAB, cerium chloride (CeCl3 and 2’, 7’-dichlorofluorescin diacetate (H2DCF-DA, to penetrate those organs. Theoretically, the reaction of endogenous H2O2 with these chemicals could be facilitated by using thin organ sections. However, the rapid production of wound-induced H2O2 associated with this procedure inevitably disturbs the original distribution of H2O2 in vivo. Here, by employing tomato seedling stems and fruits as testing materials, we report a novel, simple and rapid protocol to localize H2O2 in those organs using DAB-mediated tissue printing. The rapidity of the protocol (within 15 s completely avoided the interference of wound-induced H2O2 during experimentation. Moreover, the H2O2 signal on the printing was stable for at least 1 h with no or little background produced. We conclude that DAB-mediated tissue printing developed here provide a new feasible and reliable method to localize H2O2 in large plant organs, hence should have broad applications in studying ROS biology.

  18. A rapid and reliable procedure for extraction of cellular polyamines and inorganic ions from plant tissues

    Science.gov (United States)

    Rakesh Minocha; Walter C. Shortle; Stephanie L. Long; Subhash C. Minocha

    1994-01-01

    A fast and reliable method for the extraction of cellular polyamines and major inorganic ions (Ca, Mg, Mn, K, and P) from several plant tissues is described. The method involves repeated freezing and thawing of samples instead of homogenization. The efficiency of extraction of both the polyamines and inorganic ions by these two methods was compared for 10 different...

  19. COLORFUL-Circuit: a platform for rapid multigene assembly, delivery and expression in plants

    Directory of Open Access Journals (Sweden)

    Hassan eGhareeb

    2016-03-01

    Full Text Available Advancing basic and applied plant research requires the continuous innovative development of the available technology toolbox. Essential components of this toolbox are methods that simplify the assembly, delivery and expression of multiple transgenes of interest. To allow simultaneous and directional multigene assembly on the same plant transformation vector, several strategies based on overlapping sequences or restriction enzymes have recently been developed. However, the assembly of homologous and repetitive DNA sequences can be inefficient and the frequent occurrence of target sequences recognized by commonly used restriction enzymes can be a limiting factor. Here, we noted that recognition sites for the restriction enzyme SfiI are rarely occurring in plant genomes. This fact was exploited to establish a multigene assembly system called COLORFUL-Circuit. To this end, we developed a set of binary vectors which provide a flexible and cost efficient cloning platform. The gene expression cassettes in our system are flanked with unique SfiI sites, which allow simultaneous multi-gene cassette assembly in a hosting binary vector. We used COLORFUL-Circuit to transiently and stably express up to four fluorescent organelle markers in addition to a selectable marker and analyzed the impact of assembly design on coexpression efficiency. Finally, we demonstrate the utility of our optimized COLORFUL-Circuit system in an exemplary case study, in which we monitored simultaneously the subcellular behavior of multiple organelles in a biotrophic plant-microbe interaction by Confocal Laser Scanning Microscopy.

  20. A rapid HPLC-APCI-MS method to detect fluoroacetate in plants

    Science.gov (United States)

    Many plant species worldwide can cause sudden death of grazing livestock. One diagnostic differential is the presence of monofluoroacetate (MFA) that is metabolised to fluorocitrate that subsequently inhibits the Kreb’s Cycle (the tricarboxylic acid cycle) leading to cellular respiration dysfunction...

  1. Rapid and accurate analyses of silicon and phosphorus in plants using a portable X-ray fluorescence spectrometer.

    Science.gov (United States)

    Reidinger, Stefan; Ramsey, Michael H; Hartley, Susan E

    2012-08-01

    The elemental analysis of plant material is a frequently employed tool across biological disciplines, yet accurate, convenient and economical methods for the determination of some important elements are currently lacking. For instance, digestion-based techniques are often hazardous and time-consuming and, particularly in the case of silicon (Si), can suffer from low accuracy due to incomplete solubilization and potential volatilization, whilst other methods may require large, expensive and specialised equipment. Here, we present a rapid, safe and accurate procedure for the simultaneous, nonconsumptive analysis of Si and phosphorus (P) in as little as 0.1 g dried and ground plant material using a portable X-ray fluorescence spectrometer (P-XRF). We used certified reference materials from different plant species to test the analytical performance of P-XRF and show that the analysis suffers from very little bias and that the repeatability precision of the measurements is as good as or better than that of other methods. Using this technique we were able to process and analyse 200 ground samples a day, so P-XRF could provide a particularly valuable tool for plant biologists requiring the simultaneous nonconsumptive analysis of multiple elements, including those known to be difficult to measure such as Si, in large numbers of samples. © 2012 The Authors. New Phytologist © 2012 New Phytologist Trust.

  2. Periodontal regeneration.

    Science.gov (United States)

    Ivanovski, S

    2009-09-01

    The ultimate goal of periodontal therapy is the regeneration of the tissues destroyed as a result of periodontal disease. Currently, two clinical techniques, based on the principles of "guided tissue regeneration" (GTR) or utilization of the biologically active agent "enamel matrix derivative" (EMD), can be used for the regeneration of intrabony and Class II mandibular furcation periodontal defects. In cases where additional support and space-making requirements are necessary, both of these procedures can be combined with a bone replacement graft. There is no evidence that the combined use of GTR and EMD results in superior clinical results compared to the use of each material in isolation. Great variability in clinical outcomes has been reported in relation to the use of both EMD and GTR, and these procedures can be generally considered to be unpredictable. Careful case selection and treatment planning, including consideration of patient, tooth, site and surgical factors, is required in order to optimize the outcomes of treatment. There are limited data available for the clinical effectiveness of other biologically active molecules, such as growth factors and platelet concentrates, and although promising results have been reported, further clinical trials are required in order to confirm their effectiveness. Current active areas of research are centred on tissue engineering and gene therapy strategies which may result in more predictable regenerative outcomes in the future.

  3. Pilot plant study of alternative filter media for rapid gravity filtration.

    Science.gov (United States)

    Davies, P D; Wheatley, A D

    2012-01-01

    Sand has been the main filter media used in rapid gravity filtration since its introduction. The dominance of sand has been due to its low cost and availability. Extensive experience has led to sand filters with a dependable and predictable performance. Sand remains the preferred filter medium but usually with a larger sized anthracite capping to reduce the onset of head loss. Other approved filter media are now commercially available and this paper compares sand with recycled glass, Filtralite(®) and slate at pilot scale. The results have reaffirmed the basic importance of particle size on head loss and turbidity performance rather than surface activity or specific surface area. The results did suggest, however, that particle shape and packing exerted a stronger influence on performance than previously acknowledged. These could be used to improve the design and the contribution to sustainability made by rapid gravity filters.

  4. Interagency partnering for weed prevention--progress on development of a National Early Detection and Rapid Response System for Invasive Plants in the United States

    Science.gov (United States)

    Westbrooks, R.; Westbrooks, R.

    2011-01-01

    Over the past 50 years, experience has shown that interagency groups provide an effective forum for addressing various invasive species issues and challenges on multiple land units. However, more importantly, they can also provide a coordinated framework for early detection, reporting, identification and vouchering, rapid assessment, and rapid response to new and emerging invasive plants in the United States. Interagency collaboration maximizes the use of available expertise, resources, and authority for promoting early detection and rapid response (EDRR) as the preferred management option for addressing new and emerging invasive plants. Currently, an interagency effort is underway to develop a National EDRR System for Invasive Plants in the United States. The proposed system will include structural and informational elements. Structural elements of the system include a network of interagency partner groups to facilitate early detection and rapid response to new invasive plants, including the Federal Interagency Committee for the Management of Noxious and Exotic Weeds (FICMNEW), State Invasive Species Councils, State Early Detection and Rapid Response Coordinating Committees, State Volunteer Detection and Reporting Networks, Invasive Plant Task Forces, and Cooperative Weed Management Areas. Informational elements and products being developed include Regional Invasive Plant Atlases, and EDRR Guidelines for EDRR Volunteer Network Training, Rapid Assessment and Rapid Response, and Criteria for Selection of EDRR Species. System science and technical support elements which are provided by cooperating state and federal scientists, include EDRR guidelines, training curriculum for EDRR volunteers and agency field personnel, plant identification and vouchering, rapid assessments, as well as predictive modeling and ecological range studies for invasive plant species.

  5. Somatic Embryogenesis and Plant Regeneration in Sapindus mukorossi Gaertn. from Leaf-Derived Callus Induced with 6-Benzylaminopurine.

    Science.gov (United States)

    Singh, Reetika; Rai, Manoj Kumar; Kumari, Nishi

    2015-09-01

    A somatic embryogenesis system was developed for Sapindus mukorossi Gaertn. from leaf explants obtained from fresh flushes of a mature tree. Callus was induced from the midrib region of leaf explants on Murashige and Skoog (MS) medium containing different concentrations of 2,4-dichlorophenoxyacetic acid or 6-benzylaminopurine. Callus induction and somatic embryogenesis was significantly influenced by the size, physiological age, and orientation of leaf explants on the culture medium and plant growth regulators. Adaxial-side-up orientation of leaf explants significantly promoted embryogenesis in comparison with abaxial-side-up orientation. Maximum number of somatic embryos was induced on MS medium supplemented with 8.88 μM 6-benzylaminopurine. Scanning electron microscopy of embryogenic callus revealed somatic embryo origin and the development of globular-, heart-, and cotyledonary-stage somatic embryos. The frequency of maturation as well as germination of somatic embryos was higher on MS medium containing 8.88 μM 6-benzylaminopurine than on medium without 6-benzylaminopurine. Plantlets which developed from somatic embryos were acclimatized successfully with 90 % survival.

  6. Portable plant chlorophyll fluorimeter based on blue LED rapid induced technology

    Science.gov (United States)

    Zheng, Yibo; Mi, Ting; Zhang, Lei; Zhao, Jun

    2018-01-01

    Fluorimeter is an effective device for detecting chlorophyll a content in plants. In order to realize real-time nondestructive detection of plant blades, a camera based fluorescence instrument based on two color mirrors has been developed. The blue light LED is used as the excitation light source, and the lens is used for shaping and focusing the excitation light to ensure the excitation intensity and uniform illumination of the light source. The device uses a 45 degree two color mirror to separate the chlorophyll a excited light path and the fluorescence receiving light path. Finally, the fluorescent signal is collected by the silicon photocell, and the signal is processed by the circuit to transmit the digital information to the display. Through the analysis of the experimental data, the device has the advantages of small size, easy to carry, fast induction, etc., and can be widely applied in outdoor teaching and field investigation.

  7. [Establishment of embryogenic cell suspension culture and plant regeneration of edible banana Musa acuminata cv. Mas (AA)].

    Science.gov (United States)

    Wei, Yue-Rong; Huang, Xue-Lin; Li, Jia; Huang, Xia; Li, Zhe; Li, Xiao-Ju

    2005-01-01

    Conventional breeding for dual resistance of disease and pest of Musa cultivars remains a difficult endeavor, as the plant is polyploidic and high in sterility. Biotechnological techniques, eg., genetic engineering, in vitro mutation breeding, or protoplast fusion, may overcome the difficulties and improve the germplasm. Establishment of a stable embryogenic cell suspension (ECS) is a prerequisite for any of the biotechnological breeding methods. In this study an embryogenic cell suspension was established from immature male flower of Musa acuminata cv. Mas (AA), a popular commercial variety of banana in the South-East Asian region. After culture for 5-6 months on callus induction media, which consisted of MS salts, different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D), 4.1 micromol/L biotin, 5.7 micromol/L indoleacetic acid (IAA), 5.4 micromol/L naphthaleneacetic acid (NAA), other vitamins, 87 mmol/L sucrose, and solidified with 7 g/L agarose, meristematic globules and yellow, friable embryogenic cultures were induced from the explants of 1-15th row young floral hands of immature male flowers. Of the four treatments of 2,4-D, 9 micromol/L was the most effective on the callus induction, it transformed 40.96% and 7.45% of the cultivated male floral hands into callus and embryogenic callus respectively. The explants to produce highest frequency of the embryogenic calli were floral hands of 6 to 12th rows, which generated 5.79% of the embryogenic calli. Suspension cultures were initiated from these embryogenic calli in liquid medium supplemented with 4.5 micromol/L 2, 4-D. After sieving selection of the cultures using a stainless steel metallic strainer with pore sizes of 154 microm at 15 day intervals for 3 months, homogeneous and yellow embryogenic cell suspensions, composed of single cells and small cell aggregates, were established. Based upon the growth quantity and growth rate of ECS, it was determined that the appropriate inoculum was 2.0 mL PCV

  8. Rapid atmospheric transport and large-scale deposition of recently synthesized plant waxes

    Science.gov (United States)

    Nelson, Daniel B.; Ladd, S. Nemiah; Schubert, Carsten J.; Kahmen, Ansgar

    2018-02-01

    Sedimentary plant wax 2H/1H ratios are important tools for understanding hydroclimate and environmental changes, but large spatial and temporal uncertainties exist about transport mechanisms from ecosystem to sediments. To assess atmospheric pathways, we collected aerosol samples for two years at four locations within a ∼60 km radius in northern Switzerland. We measured n-alkane distributions and 2H/1H ratios in these samples, and from local plants, leaf litter, and soil, as well as surface sediment from six nearby lakes. Increased concentrations and 2H depletion of long odd chain n-alkanes in early summer aerosols indicate that most wax aerosol production occurred shortly after leaf unfolding, when plants synthesize waxes in large quantities. During autumn and winter, aerosols were characterized by degraded n-alkanes lacking chain length preferences diagnostic of recent biosynthesis, and 2H/1H values that were in some cases more than 100‰ higher than growing season values. Despite these seasonal shifts, modeled deposition-weighted average 2H/1H values of long odd chain n-alkanes primarily reflected summer values. This was corroborated by n-alkane 2H/1H values in lake sediments, which were similar to deposition-weighted aerosol values at five of six sites. Atmospheric deposition rates for plant n-alkanes on land were ∼20% of accumulation rates in lakes, suggesting a role for direct deposition to lakes or coastal oceans near similar production sources, and likely a larger role for deposition on land and transport in river systems. This mechanism allows mobilization and transport of large quantities of recently produced waxes as fine-grained material to low energy sedimentation sites over short timescales, even in areas with limited topography. Widespread atmospheric transfer well before leaf senescence also highlights the importance of the isotopic composition of early season source water used to synthesize waxes for the geologic record.

  9. Plant Lesions Promote the Rapid Multiplication of Escherichia coli O157:H7 on Postharvest Lettuce▿

    Science.gov (United States)

    Brandl, M. T.

    2008-01-01

    Several outbreaks of Escherichia coli O157:H7 infections have been associated with minimally processed leafy vegetables in the United States. Harvesting and processing cause plant tissue damage. In order to assess the role of plant tissue damage in the contamination of leafy greens with E. coli O157:H7, the effect of mechanical, physiological, and plant disease-induced lesions on the growth of this pathogen on postharvest romaine lettuce was investigated. Within only 4 h after inoculation, the population sizes of E. coli O157:H7 increased 4.0-, 4.5-, and 11.0-fold on lettuce leaves that were mechanically bruised, cut into large pieces, and shredded into multiple pieces, respectively. During the same time, E. coli O157:H7 population sizes increased only twofold on leaves that were left intact after harvest. Also, the population size of E. coli O157:H7 was 27 times greater on young leaves affected by soft rot due to infection by Erwinia chrysanthemi than on healthy middle-aged leaves. Confocal microscopy revealed that leaf tip burn lesions, which are caused by a common physiological disorder of lettuce, harbored dense populations of E. coli O157:H7 cells both internally and externally. Investigation of the colonization of cut lettuce stems by E. coli O157:H7 showed that the pathogen grew 11-fold over 4 h of incubation after its inoculation onto the stems, from which large amounts of latex were released. The results of this study indicate that plant tissue damage of various types can promote significant multiplication of E. coli O157:H7 over a short time and suggest that harvesting and processing are critical control points in the prevention or reduction of E. coli O157:H7 contamination of lettuce. PMID:18641153

  10. Plant lesions promote the rapid multiplication of Escherichia coli O157:H7 on postharvest lettuce.

    Science.gov (United States)

    Brandl, M T

    2008-09-01

    Several outbreaks of Escherichia coli O157:H7 infections have been associated with minimally processed leafy vegetables in the United States. Harvesting and processing cause plant tissue damage. In order to assess the role of plant tissue damage in the contamination of leafy greens with E. coli O157:H7, the effect of mechanical, physiological, and plant disease-induced lesions on the growth of this pathogen on postharvest romaine lettuce was investigated. Within only 4 h after inoculation, the population sizes of E. coli O157:H7 increased 4.0-, 4.5-, and 11.0-fold on lettuce leaves that were mechanically bruised, cut into large pieces, and shredded into multiple pieces, respectively. During the same time, E. coli O157:H7 population sizes increased only twofold on leaves that were left intact after harvest. Also, the population size of E. coli O157:H7 was 27 times greater on young leaves affected by soft rot due to infection by Erwinia chrysanthemi than on healthy middle-aged leaves. Confocal microscopy revealed that leaf tip burn lesions, which are caused by a common physiological disorder of lettuce, harbored dense populations of E. coli O157:H7 cells both internally and externally. Investigation of the colonization of cut lettuce stems by E. coli O157:H7 showed that the pathogen grew 11-fold over 4 h of incubation after its inoculation onto the stems, from which large amounts of latex were released. The results of this study indicate that plant tissue damage of various types can promote significant multiplication of E. coli O157:H7 over a short time and suggest that harvesting and processing are critical control points in the prevention or reduction of E. coli O157:H7 contamination of lettuce.

  11. Factors influencing rapid clonal propagation of Chlorophytum arundinaceum (Liliales: Liliaceae), an endangered medicinal plant

    OpenAIRE

    Sanghamitra Samantaray; Satyabrata Maiti

    2011-01-01

    Chlorophytum arundinaceum is an important medicinal plant and its tuberous roots are used for various health ailment treatments. It has become an endangered species in the Eastern Ghats, and a rare medicinal herb in India, due to its excessive collection from its natural habitat and its destructive harvesting techniques, coupled with poor seed germination and low vegetative multiplication ratio. In order to contribute to its production systems, an efficient protocol was developed for in vitro...

  12. Adjustment of a rapid method for quantification of Fusarium spp. spore suspensions in plant pathology.

    Science.gov (United States)

    Caligiore-Gei, Pablo F; Valdez, Jorge G

    2015-01-01

    The use of a Neubauer chamber is a broadly employed method when cell suspensions need to be quantified. However, this technique may take a long time and needs trained personnel. Spectrophotometry has proved to be a rapid, simple and accurate method to estimate the concentration of spore suspensions of isolates of the genus Fusarium. In this work we present a linear formula to relate absorbance measurements at 530nm with the number of microconidia/ml in a suspension. Copyright © 2014 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  13. Artemisinin production by shoot regeneration of Artemisia annua L. using thidiazuron.

    Science.gov (United States)

    Lualon, Wanwimon; De-Eknamkul, Wanchai; Tanaka, Hiroyuki; Shoyama, Yukihiro; Putalun, Waraporn

    2008-01-01

    An efficient in vitro method for multiple shoot bud induction and regeneration has been developed in Artemisia annua L. using leaf and stem explants in various concentrations and combinations of plant growth regulators to evaluate the frequency of regeneration. The sources of explants as well as plant growth regulators in the medium were found to influence the multiple shoot induction. The result shows that the stem segment cultured on Murashige and Skoog (MS) medium supplemented with 0.1 mg/l thidiazuron (TDZ) gave a perfect shoot formation (100%) and good shoot multiplication (57 shoots/explant) after 2 weeks of culture. Healthy regenerated shoots were elongated and rooted in MS medium without hormones. The artemisinin content in plants regenerated from stem explants using 0.1 mg/l TDZ was (3.36 +/- 0.36) microg/mg dry weight and two-fold higher than that of in vitro grown plants of the same age [(1.73 -/+ 0.23) microg/mg DW]. This system exhibited a potential for a rapid propagation of shoots from the stem explant and makes it possible to develop a clonal propagation of A. annua.

  14. Impact of application of zinc oxide nanoparticles on callus induction, plant regeneration, element content and antioxidant enzyme activity in tomato (Solanum lycopersicum Mill. under salt stress

    Directory of Open Access Journals (Sweden)

    Alharby Hesham F.

    2016-01-01

    Full Text Available The properties of nanomaterials and their potential applications have been given considerable attention by researchers in various fields, especially agricultural biotechnology. However, not much has been done to evaluate the role or effect of zinc oxide nanoparticles (ZnO-NP in regulating physiological and biochemical processes in response to salt-induced stress. For this purpose, some callus growth traits, plant regeneration rate, mineral element (sodium, potassium, phosphorous and nitrogen contents and changes in the activity of superoxide dismutase (SOD and glutathione peroxidase (GPX in tissues of five tomato cultivars were investigated in a callus culture exposed to elevated concentrations of salt (3.0 and 6.0 g L-1NaCl, and in the presence of zinc oxide nanoparticles (15 and 30 mg L-1. The relative callus growth rate was inhibited by 3.0 g L-1 NaCl; this was increased dramatically at 6.0 g L-1. Increasing exposure to NaCl was associated with a significantly higher sodium content and SOD and GPX activities. Zinc oxide nanoparticles mitigated the effects of NaCl, and in this application of lower concentrations (15 mg L-1 was more effective than a higher concentration (30 mg L-1. This finding indicates that zinc oxide nanoparticles should be investigated further as a potential anti-stress agent in crop production. Different tomato cultivars showed different degrees of tolerance to salinity in the presence of ZnO-NP. The cultivars Edkawy, followed by Sandpoint, were less affected by salt stress than the cultivar Anna Aasa.

  15. Arabidopsis seedling flood-inoculation technique: a rapid and reliable assay for studying plant-bacterial interactions

    Directory of Open Access Journals (Sweden)

    Uppalapati Srinivasa R

    2011-10-01

    Full Text Available Abstract Background The Arabidopsis thaliana-Pseudomonas syringae model pathosystem is one of the most widely used systems to understand the mechanisms of microbial pathogenesis and plant innate immunity. Several inoculation methods have been used to study plant-pathogen interactions in this model system. However, none of the methods reported to date are similar to those occurring in nature and amicable to large-scale mutant screens. Results In this study, we developed a rapid and reliable seedling flood-inoculation method based on young Arabidopsis seedlings grown on MS medium. This method has several advantages over conventional soil-grown plant inoculation assays, including a shorter growth and incubation period, ease of inoculation and handling, uniform infection and disease development, requires less growth chamber space and is suitable for high-throughput screens. In this study we demonstrated the efficacy of the Arabidopsis seedling assay to study 1 the virulence factors of P. syringae pv. tomato DC3000, including type III protein secretion system (TTSS and phytotoxin coronatine (COR; 2 the effector-triggered immunity; and 3 Arabidopsis mutants affected in salicylic acid (SA- and pathogen-associated molecular pattern (PAMPs-mediated pathways. Furthermore, we applied this technique to study nonhost resistance (NHR responses in Arabidopsis using nonhost pathogens, such as P. syringae pv. tabaci, pv. glycinea and pv. tomato T1, and confirmed the functional role of FLAGELLIN-SENSING 2 (FLS2 in NHR. Conclusions The Arabidopsis seedling flood-inoculation assay provides a rapid, efficient and economical method for studying Arabidopsis-Pseudomonas interactions with minimal growth chamber space and time. This assay could also provide an excellent system for investigating the virulence mechanisms of P. syringae. Using this method, we demonstrated that FLS2 plays a critical role in conferring NHR against nonhost pathovars of P. syringae, but not to

  16. Multichannel microfluidic chip for rapid and reliable trapping and imaging plant-parasitic nematodes

    Science.gov (United States)

    Amrit, Ratthasart; Sripumkhai, Witsaroot; Porntheeraphat, Supanit; Jeamsaksiri, Wutthinan; Tangchitsomkid, Nuchanart; Sutapun, Boonsong

    2013-05-01

    Faster and reliable testing technique to count and identify nematode species resided in plant roots is therefore essential for export control and certification. This work proposes utilizing a multichannel microfluidic chip with an integrated flow-through microfilter to retain the nematodes in a trapping chamber. When trapped, it is rather simple and convenient to capture images of the nematodes and later identify their species by a trained technician. Multiple samples can be tested in parallel using the proposed microfluidic chip therefore increasing number of samples tested per day.

  17. Rapid evolution and range expansion of an invasive plant are driven by provenance-environment interactions.

    Science.gov (United States)

    Zenni, Rafael D; Bailey, Joseph K; Simberloff, Daniel

    2014-06-01

    To improve our ability to prevent and manage biological invasions, we must understand their ecological and evolutionary drivers. We are often able to explain invasions after they happen, but our predictive ability is limited. Here, we show that range expansions of introduced Pinus taeda result from an interaction between genetic provenance and climate and that temperature and precipitation clines predict the invasive performance of particular provenances. Furthermore, we show that genotypes can occupy climate niche spaces different from those observed in their native ranges and, at least in our case, that admixture is not a main driver of invasion. Genotypes respond to climate in distinct ways, and these interactions affect the ability of populations to expand their ranges. While rapid evolution in introduced ranges is a mechanism at later stages of the invasion process, the introduction of adapted genotypes is a key driver of naturalisation of populations of introduced species. © 2014 John Wiley & Sons Ltd/CNRS.

  18. Rapid estimation of nutrients in chicken manure during plant-field composting using physicochemical properties.

    Science.gov (United States)

    Huang, Guangqun; Wang, Xiaoyan; Han, Lujia

    2011-01-01

    Regression equations which relate livestock and poultry manure nutrient content to its several physicochemical properties have been reported by previous researchers. This study explores the feasibility and efficiency to determine the nutrients (TN; TP; TK; Cu and Zn) in chicken manure during composting using physicochemical properties (pH, EC and DM), and compares the performances of regression equations in this study with those in the literature. The results show that DM is the best predictor to construct the single linear regressions for all the nutrients (R2≥0.84, p<0.001). In addition, the multiple linear regression equations based on DM and pH are all notable. These findings show the potential of physicochemical models for TN, TP, TK, Cu and Zn with more convenience and rapidness, but further research is needed to develop better models with higher accuracy for the above and other more nutrients. Copyright © 2010 Elsevier Ltd. All rights reserved.

  19. Flower litters of alpine plants affect soil nitrogen and phosphorus rapidly in the eastern Tibetan Plateau

    Science.gov (United States)

    Wang, Jinniu; Xu, Bo; Wu, Yan; Gao, Jing; Shi, Fusun

    2016-10-01

    Litters of reproductive organs have rarely been studied despite their role in allocating nutrients for offspring reproduction. This study determines the mechanism through which flower litters efficiently increase the available soil nutrient pool. Field experiments were conducted to collect plant litters and calculate biomass production in an alpine meadow of the eastern Tibetan Plateau. C, N, P, lignin, cellulose content, and their relevant ratios of litters were analyzed to identify their decomposition features. A pot experiment was performed to determine the effects of litter addition on the soil nutrition pool by comparing the treated and control samples. The litter-bag method was used to verify decomposition rates. The flower litters of phanerophyte plants were comparable with non-flower litters. Biomass partitioning of other herbaceous species accounted for 10-40 % of the aboveground biomass. Flower litter possessed significantly higher N and P levels but less C / N, N / P, lignin / N, and lignin and cellulose concentrations than leaf litter. The litter-bag experiment confirmed that the flower litters of Rhododendron przewalskii and Meconopsis integrifolia decompose approximately 3 times faster than mixed litters within 50 days. Pot experiment findings indicated that flower litter addition significantly increased the available nutrient pool and soil microbial productivity. The time of litter fall significantly influenced soil available N and P, and soil microbial biomass. Flower litters fed the soil nutrition pool and influenced nutrition cycling in alpine ecosystems more efficiently because of their non-ignorable production, faster decomposition rate, and higher nutrient contents compared with non-flower litters. The underlying mechanism can enrich nutrients, which return to the soil, and non-structural carbohydrates, which feed and enhance the transitions of soil microorganisms.

  20. Rapid Growth and Apparent Total Nitrogen Increases in Rice and Corn Plants following Applications of Triacontanol.

    Science.gov (United States)

    Knowles, N R; Ries, S K

    1981-12-01

    Triacontanol (TRIA) increased fresh and dry weight and total reducible nitrogen (total N) of rice (Oryza sativa L.) seedlings within 40 minutes. Increases in total N in the supernatants from homogenates of corn (Zea mays L.) and rice leaves treated with TRIA for one minute before grinding occurred within 30 and 80 minutes, respectively. The source for the increase was investigated utilizing atmospheric substitution and enrichment and depletion studies with (15)N. The increase in total N in seedlings was shown to be independent of method of N analysis and the presence of nitrate in the plants. Automated Kjeldahl determinations showing apparent increases in N composition due to TRIA were shown to be correlated with hand Kjeldahl, elemental analysis, and chemiluminescent analysis in three independent laboratories. TRIA did not alter the nitrate uptake or endogenous levels of nitrate in corn and rice seedlings. Enrichment experiments revealed that the total N increases in rice seedlings, in vivo, and in supernatants of corn leaf homogenates, in vitro, are not due to atmospheric N(2). TRIA increased the soluble N pools of the plants, specifically the free amino acid and soluble protein fractions. No differences in depletion or enrichment of (15)N incorporated into soluble and insoluble N fractions of rice seedlings could be detected on an atom per cent (15)N basis. The apparent short-term total N increases cannot be explained by current knowledge of major N assimilation pathways. TRIA may stimulate a change in the chemical composition of the seedlings, resulting in interference with standard methods of N analysis.

  1. Application of barcode high-resolution melting for rapid authentication of the medicinal plant Psammosilene tunicoides

    Directory of Open Access Journals (Sweden)

    Jingjian Li

    2016-07-01

    Full Text Available Psammosilene tunicoides is an important herb used in traditional Chinese medicine. It has been proved to benefit the stomach and treat rheumatism and gout. On the market, this herbal medicine is frequently adulterated by the related species Silene viscidula. Correct identification of P. tunicoides is important to ensure herbal quality, safety, authenticity and health for consumers. However, the identification of the P. tunicoides and S. viscidula species is complicated because of their morphological similarities. Therefore, a reliable method to authenticate these two medicinal plants is needed. In this study, the ITS2 barcode region coupled with high-resolution melting (Bar-HRM was evaluated as a novel approach for differentiating P. tunicoides from its adulterant S. viscidula. Our findings showed that Bar-HRM not only detected the adulteration, but also quantified the most common admixture. Bar-HRM sensitivity in adulterant detection was assessed by analysing samples mixed with different proportions of S. viscidula and P. tunicoides control. The results are presented as a linear regression with R2 = 0.9852, which implied the capability of the method to detect adulteration. This study is significant to verify the authenticity for better quality control of this herbal species.

  2. Ball mill assisted rapid mechanochemical extraction method for natural products from plants.

    Science.gov (United States)

    Wang, Man; Bi, Wentao; Huang, Xiaohua; Chen, David Da Yong

    2016-06-03

    A ball mill assisted mechanochemical extraction method was developed to extract compounds of natural product (NP) from plant using ionic liquid (IL). A small volume ball mill, also known as PastPrep(®) Homogenizer, which is often used for high-speed lysis of biological samples and for other applications, was used to dramatically increase the speed, completeness and reproducibility of the extraction process at room temperature to preserve the chemical integrity of the extracted compounds. In this study, tanshinones were selected as target compounds to evaluate the performance of this extraction method. Factors affecting the extraction efficiency, such as the duration, IL concentration and solid/liquid ratio were systematically optimized using the response surface methodology. Under the optimized conditions, the described method was more efficient and much faster than the conventional extraction methods such as methanol based ultrasound assisted extraction (UAE) and heat reflux extraction (HRE) that consumes a lot more organic solvent. In addition, the natural products of interest were enriched by anion metathesis of ionic liquids, combining extraction and preconcentration in the same process. The extractant was analyzed by HPLC and LC-MS. The reproducibility (RSD, n=5), correlation coefficient (r(2)) of the calibration curve, and the limit of detection, were determined to be in the range of 4.7-5.2%, 0.9992-0.9995, and 20-51ng/mL, respectively. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Interaction rewiring and the rapid turnover of plant-pollinator networks.

    Science.gov (United States)

    CaraDonna, Paul J; Petry, William K; Brennan, Ross M; Cunningham, James L; Bronstein, Judith L; Waser, Nickolas M; Sanders, Nathan J

    2017-03-01

    Whether species interactions are static or change over time has wide-reaching ecological and evolutionary consequences. However, species interaction networks are typically constructed from temporally aggregated interaction data, thereby implicitly assuming that interactions are fixed. This approach has advanced our understanding of communities, but it obscures the timescale at which interactions form (or dissolve) and the drivers and consequences of such dynamics. We address this knowledge gap by quantifying the within-season turnover of plant-pollinator interactions from weekly censuses across 3 years in a subalpine ecosystem. Week-to-week turnover of interactions (1) was high, (2) followed a consistent seasonal progression in all years of study and (3) was dominated by interaction rewiring (the reassembly of interactions among species). Simulation models revealed that species' phenologies and relative abundances constrained both total interaction turnover and rewiring. Our findings reveal the diversity of species interactions that may be missed when the temporal dynamics of networks are ignored. © 2017 John Wiley & Sons Ltd/CNRS.

  4. Recent origin and rapid speciation of Neotropical orchids in the world's richest plant biodiversity hotspot.

    Science.gov (United States)

    Pérez-Escobar, Oscar Alejandro; Chomicki, Guillaume; Condamine, Fabien L; Karremans, Adam P; Bogarín, Diego; Matzke, Nicholas J; Silvestro, Daniele; Antonelli, Alexandre

    2017-07-01

    The Andean mountains of South America are the most species-rich biodiversity hotspot worldwide with c. 15% of the world's plant species, in only 1% of the world's land surface. Orchids are a key element of the Andean flora, and one of the most prominent components of the Neotropical epiphyte diversity, yet very little is known about their origin and diversification. We address this knowledge gap by inferring the biogeographical history and diversification dynamics of the two largest Neotropical orchid groups (Cymbidieae and Pleurothallidinae), using two unparalleled, densely sampled orchid phylogenies (including more than 400 newly generated DNA sequences), comparative phylogenetic methods, geological and biological datasets. We find that the majority of Andean orchid lineages only originated in the last 20-15 million yr. Andean lineages are derived from lowland Amazonian ancestors, with additional contributions from Central America and the Antilles. Species diversification is correlated with Andean orogeny, and multiple migrations and recolonizations across the Andes indicate that mountains do not constrain orchid dispersal over long timescales. Our study sheds new light on the timing and geography of a major Neotropical diversification, and suggests that mountain uplift promotes species diversification across all elevational zones. © 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.

  5. [Rapid determination of eight organic acids in plant tissue by sequential extraction and high performance liquid chromatography].

    Science.gov (United States)

    Huang, Tianzhi; Wang, Shijie; Liu Xiuming; Liu, Hong; Wu, Yanyou; Luo Xuqiang

    2014-12-01

    A sequential extraction method was developed to determine different forms of oxalate and seven oxalate-metabolism-related organic acids (glyoxylic acid, tartaric acid, glycolic acid, malic acid, acetic acid, citric acid, succinic acid) in plant tissue. The ultra-pure water was used as the extraction medium to obtain water-soluble oxalic acid and the other seven water-soluble organic acids. After the extraction of the water-soluble organic acids, the residues were extracted by dilute hydrochloric acid successively to get the acid-soluble oxalate which entered the liquid phase. A Hypersil ODS column was used with 5 mmol/L potassium dihydrogen phosphate buffer solution (pH 2. 8) as the mobile phase. The diode array detector was set at 210 nm and the column temperature at 30 °C with the injection volume of 5 µL. The flow rate was controlled at different times which allowed a good and rapid separation of the organic acids and hydrochloric acid. Under these conditions, the linear ranges of the method were 1-2000 mg/L for oxalic acid, 25-2,000 mg/L for acetic acid, and 10-2,000 mg/L for glyoxylic acid, tartaric acid, glycolic acid, malic acid, citric acid and succinic acid, with the correlation coefficients of the eight organic acids ≥ 0. 9996. The average recoveries of the eight organic acids in leaves and roots were 93. 5%-104. 4% and 85. 3%-105. 4% with RSDs of 0. 15% -2.43% and 0. 31%-2. 9% (n=7), respectively. The limits of detection ranged from 1 to 10 ng (S/N=3). The results indicated that the method is accurate, rapid and reproducible for the determination of organic acids in plant samples.

  6. Bleach Plant Capital Reduction with Rapid DO Bleaching and Simplified (D/E/D) Stages

    Energy Technology Data Exchange (ETDEWEB)

    T. J. McDonough; C. E. Courchene; J-C. Baromes

    2000-08-01

    The objective of this work was to demonstrate the capabilities of a bleaching sequence that combined a short retention time initial chlorine dioxide stage, referred to as rapid D0, (D0R), with simplified bleaching stages, (D1/E/D2), that required only one final bleach washer. The test sequence DR(EPO)(D/E/D/) was compared to a control sequence, D(EPO)D, for both hardwood and softwood pulps. The capabilities of the DR(EPO)(D/E/D) sequence were successfully demonstrated. An existing three- or four-stage bleach plan can be converted to the more powerful DR(EPO)(D/E/D) sequence without the major capital cost of additional washers. The results from this study showed that the DR(EPO)(D/E/D) sequence can reach 85 brightness on SW with 2.8% total C1O2, while the control sequence, D(EPO)D, required 3.9% C1O2. There was a corresponding decrease in AOX for the test sequence. The strength of pulp bleached in the test sequence was similar to or slightly higher than the control. For the HW pu lp, the test sequence reached 88 brightness with 2.2% C1O2 compared to 3.3% C1O2 for the control. There was a corresponding decrease in AOX generation with the lower chemical requirements. The final viscosity and pulp strength for the test sequence on HW was significantly higher than the corresponding values for the control sequence.

  7. Factors influencing rapid clonal propagation of Chlorophytum arundinaceum (Liliales: Liliaceae, an endangered medicinal plant

    Directory of Open Access Journals (Sweden)

    Sanghamitra Samantaray

    2011-03-01

    Full Text Available Chlorophytum arundinaceum is an important medicinal plant and its tuberous roots are used for various health ailment treatments. It has become an endangered species in the Eastern Ghats, and a rare medicinal herb in India, due to its excessive collection from its natural habitat and its destructive harvesting techniques, coupled with poor seed germination and low vegetative multiplication ratio. In order to contribute to its production systems, an efficient protocol was developed for in vitro clonal propagation through shoot bud culture. For this, multiple shoots were induced from shoot bud explants on Murashige and Skoog’s medium supplemented with 2.5-3.0mg/L BAP, 0.01-0.1mg/L NAA and 3% (w/v sucrose. Inclusion of Adenine Sulphate (25mg/L in the culture medium improved the frequency of multiple shoot production and recovered the chlorotic symptoms of the leaves. Media having pH 5.9 and 4% sucrose showed significant improvement on shoot bud multiplication and growth. In vitro flowering was observed when the subcultures were carried out for over four months in the same multiplication media. Rooting was readily achieved upon transferring the shoots on to half- strength MS medium supplemented with 0.1mg/L IBA and 2% (w/v sucrose. Micropropagated plantlets were hardened in the green house, successfully established, and flowered in the field. This method could effectively be applied for the conservation and clonal propagation to meet the demand of planting materials. Rev. Biol. Trop. 59 (1: 435-445. Epub 2011 March 01.Chlorophytum arundinaceum es una planta medicinal importante y sus raíces se utilizan en diversos tratamientos contra enfermedades. Se ha convertido en una especie en peligro de extinción en el Ghats Oriental y una hierba medicinal rara en la India, debido a la recolecta excesiva en su hábitat natural y la manera destructiva de cosecharla, asociado con una mala germinación y pobre multiplicación vegetativa. Para contribuir con

  8. Rapid plant invasion in distinct climates involves different sources of phenotypic variation.

    Directory of Open Access Journals (Sweden)

    Arnaud Monty

    Full Text Available When exotic species spread over novel environments, their phenotype will depend on a combination of different processes, including phenotypic plasticity (PP, local adaptation (LA, environmental maternal effects (EME and genetic drift (GD. Few attempts have been made to simultaneously address the importance of those processes in plant invasion. The present study uses the well-documented invasion history of Senecio inaequidens (Asteraceae in southern France, where it was introduced at a single wool-processing site. It gradually invaded the Mediterranean coast and the Pyrenean Mountains, which have noticeably different climates. We used seeds from Pyrenean and Mediterranean populations, as well as populations from the first introduction area, to explore the phenotypic variation related to climatic variation. A reciprocal sowing experiment was performed with gardens under Mediterranean and Pyrenean climates. We analyzed climatic phenotypic variation in germination, growth, reproduction, leaf physiology and survival. Genetic structure in the studied invasion area was characterized using AFLP. We found consistent genetic differentiation in growth traits but no home-site advantage, so weak support for LA to climate. In contrast, genetic differentiation showed a relationship with colonization history. PP in response to climate was observed for most traits, and it played an important role in leaf trait variation. EME mediated by seed mass influenced all but leaf traits in a Pyrenean climate. Heavier, earlier-germinating seeds produced larger individuals that produced more flower heads throughout the growing season. However, in the Mediterranean garden, seed mass only influenced the germination rate. The results show that phenotypic variation in response to climate depends on various ecological and evolutionary processes associated with geographical zone and life history traits. Seeing the relative importance of EME and GD, we argue that a "local

  9. Microwave assisted extraction-solid phase extraction for high-efficient and rapid analysis of monosaccharides in plants.

    Science.gov (United States)

    Zhang, Ying; Li, Hai-Fang; Ma, Yuan; Jin, Yan; Kong, Guanghui; Lin, Jin-Ming

    2014-11-01

    Monosaccharides are the fundamental composition units of saccharides which are a common source of energy for metabolism. An effective and simple method consisting of microwave assisted extraction (MAE), solid phase extraction (SPE) and high performance liquid chromatography-refractive index detector (HPLC-RID) was developed for rapid detection of monosaccharides in plants. The MAE was applied to break down the structure of the plant cells and release the monosaccharides, while the SPE procedure was adopted to purify the extract before analysis. Finally, the HPLC-RID was employed to separate and analyze the monosaccharides with amino column. As a result, the extraction time was reduced to 17 min, which was nearly 85 times faster than soxhlet extraction. The recoveries of arabinose, xylose, fructose and glucose were 85.01%, 87.79%, 103.17%, and 101.24%, with excellent relative standard deviations (RSDs) of 1.94%, 1.13%, 0.60% and 1.67%, respectively. The proposed method was demonstrated to be efficient and time-saving, and had been applied to analyze monosaccharides in tobacco and tea successfully. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. Screening for in vitro shoot-forming capacity of seedling explants in bell pepper (Capsicum annuum L.) genotypes and efficient plant regeneration using thidiazuron.

    Science.gov (United States)

    Szász, A; Nervo, G; Fári, M

    1995-07-01

    In vitro shoot regeneration ability of 17 (7 Italian and 10 Hungarian) bell pepper genotypes was investigated using excised cotyledons and rooted hypocotyls as explants. Most of the Italian genotypes and two of the Hungarian genotypes responded well, producing shoots from rooted hypocotyls. Only two genotypes (one Italian and one Hungarian) gave a weak response using cotyledons. For direct shoot induction in these explants, in addition to the methods cited in the relevant papers, a new method was applied using thidiazuron as a cytokinin. Shoots were successfully regenerated from cotyledons of two Italian and two Hungarian genotypes using thidiazuron which were considered to be non responsive to the usual methods.

  11. Gene therapy for tissue regeneration.

    Science.gov (United States)

    Cutroneo, Kenneth R

    2003-02-01

    Tissue repair and regeneration are the normal biological responses of many different tissues in the body to injury. During the healing process, profound changes occur in cell composition and extracellular matrix (ECM) formation. Fibroblasts and equivalent reparative cells migrate to the wounded area and subsequently proliferate. These cells and reparative cells from the surrounding tissue are responsible for the rapid repair which results in tissue regeneration. Growth factors, one of which is transforming growth factor-beta (TGF-beta), stimulate fibroblasts and smooth muscle cells to proliferate and synthesize ECM proteins. This process of early repair provides a rapid way to restore new tissue and mechanical integrity. This early tissue repair process is normally followed by involution, which requires the production and activation of proteases, tissue maturation and remodeling, reorganization and finally regeneration. Alternately, failure to replace the critical components of the ECM, including elastin and basement membrane, results in abnormal regeneration of the epithelial cell layer. Although remodeling should occur during healing, provisional repair may be followed by excessive synthesis and deposition of collagen, which results in irreversible fibrosis and scarring. This excessive fibrosis which occurs in aberrant healing is at least in part mediated by persistent TGF-beta. Because of the central role of collagen in the wound healing process, the pharmacological control of collagen synthesis has been of paramount importance as a possible way to abrogate aberrant healing and prevent irreversible fibrosis. Fibrosis is an abnormal response to tissue injury. Copyright 2002 Wiley-Liss, Inc.

  12. Applied and fundamental aspects of BABY BOOM-mediated regeneration

    NARCIS (Netherlands)

    Heidmann, I.A.

    2015-01-01

    Keywords: Somatic embryogenesis, Transcription factor, AINTEGUMENTA-LIKE, BABY BOOM, BBM, Sweet Pepper Transformation Title: Applied and Fundamental Aspects of BBM-mediated Regeneration Author: Iris Heidmann Catergories: Plant regeneration, Plant

  13. Reproducible and expedient rice regeneration system using in vitro ...

    African Journals Online (AJOL)

    Yomi

    2012-01-03

    Jan 3, 2012 ... Inevitable prerequisite for expedient regeneration in rice is the selection of totipotent explant and developing an apposite combination of growth hormones. Here, we reported a reproducible regeneration protocol in which basal segments of the stem of the in vitro grown rice plants were used as ex-plant.

  14. Reproducible and expedient rice regeneration system using in vitro ...

    African Journals Online (AJOL)

    Inevitable prerequisite for expedient regeneration in rice is the selection of totipotent explant and developing an apposite combination of growth hormones. Here, we reported a reproducible regeneration protocol in which basal segments of the stem of the in vitro grown rice plants were used as ex-plant. Using the protocol ...

  15. High efficiency regeneration and genetic stability analysis of somatic ...

    African Journals Online (AJOL)

    Administrator

    2011-09-07

    Sep 7, 2011 ... Genetic stability of regenerants. In order to confirm genetic integrity, the DNA of 40 random-selected regenerated plants was compared with the DNA of the mother plant. The 36 RAPD primers used in this analysis gave rise to 150 scorable band classes, ranging from 150 bp to 2.5 kb in size. The number of.

  16. PLETHORA genes control regeneration by a two-step mechanism

    NARCIS (Netherlands)

    Kareem, Abdul; Durgaprasad, Kavya; Sugimoto, Kaoru; Du, Yujuan; Pulianmackal, Ajai J.; Trivedi, Zankhana B.; Abhayadev, Pazhoor V.; Pinon, Violaine; Meyerowitz, Elliot M.; Scheres, Ben; Prasad, Kalika

    2015-01-01

    Summary Regeneration, a remarkable example of developmental plasticity displayed by both plants and animals, involves successive developmental events driven in response to environmental cues. Despite decades of study on the ability of the plant tissues to regenerate a complete fertile shoot

  17. Restriction fragment length polymorphism of the 5S-rRNA-NTS region: a rapid and precise method for plant identification.

    Science.gov (United States)

    Bertea, Cinzia Margherita; Gnavi, Giorgio

    2012-01-01

    Molecular genetic methods have several advantages over classical morphological and chemical analyses. The genetic method requires genotype instead than phenotype, therefore PCR-based techniques have been widely used for a rapid identification of plant species, varieties and chemotypes. Recently, the molecular discrimination of some higher plant species has been evaluated using sequences of a 5S-rRNA gene spacer region. The variation in the nontranscribed sequence (NTS) region has been used in a number of plant species for studying intraspecific variation, genome evolution, and phylogenetic reconstruction. Here, we describe a rapid method based on the use of the 5S-rRNA-NTS region as a tool for plant DNA fingerprinting, which combines PCR, sequencing and restriction fragment length polymorphism analyses.

  18. Rapid compositional change and significant loss of plant species diversity among Triassic-Jurassic palynofloras in East Greenland

    Science.gov (United States)

    Mander, Luke; Kürschner, Wolfram; McElwain, Jennifer

    2010-05-01

    The Triassic-Jurassic (Tr-J; 200Ma) transition coincides with the eruption of massive flood basalts associated with the opening of the Atlantic Ocean. This is thought to have lead to a fourfold increase in palaeoatmospheric carbon dioxide, a consequent rise in global temperatures of between 3 and 6 degrees Celsius, and a rise in atmospheric pollutants such as sulphur dioxide. Recent work has employed either plant macrofossils (mostly leaves) or sporomorphs (pollen and spores) to reconstruct the response of terrestrial vegetation to this episode of major environmental change. Investigations of the macrofossil record at Astartekloft in East Greenland indicate a rapid loss of plant diversity in the Late Rhaetian, culminating in an 80% species turnover at the Tr-J boundary interval. However, evidence for such catastrophic diversity loss is conspicuously absent from the sporomorph record. This fossil group indicates that the Tr-J boundary interval in central and northwest Europe is characterized by compositional change and a transient shift from gymnosperm forests to fern-dominated vegetation. In order to address this uncertainty regarding Tr-J vegetation change according to macrofossils versus sporomorphs, we present an analysis of sporomorph diversity and compositional change across the Tr-J at Astartekloft, East Greenland. Sporomorph diversity was estimated using individual and sample-based rarefaction techniques, and compositional differences between sporomorph samples were assessed using non-metric multidimensional scaling. These analyses reveal that sporomorph assemblages from the Tr-J boundary interval at Astartekloft are between 23 and 27% less taxonomically diverse than other Triassic assemblages, and that this interval is characterized by a dramatic shift in the composition of the standing vegetation. These results are statistically significant and are also unrelated to changes in the environment of deposition. These results indicate that the magnitude of

  19. Mixture screening and mixture-amount designs to determine plant growth regulator effects on shoot regeneration from grapefruit (Citrus paradisi macf.) epicotyls

    Science.gov (United States)

    The objective of this study was to improve shoot regeneration from grapefruit epicotyl explants since some important in vitro applications in citrus, such as Agrobacterium-mediated transformation, commonly use epicotyl segments from in vitro seedlings; adequate adventitious shoot production is thus...

  20. Regeneration in Echinoderms: repair, regrowth, cloning

    OpenAIRE

    MD Candia Carnevali

    2006-01-01

    Regenerative potential is expressed to a maximum extent in echinoderms. It is a commonphenomenon in all the classes, extensively employed to reconstruct external appendages and internalorgans often subjected to amputation, self-induced or traumatic, rapidly followed by completesuccessful re-growth of the lost parts. Regeneration has been studied in adult individuals as well as inlarvae. In armed echinoderms, regeneration of arms is obviously frequent: in many cases, thedetached body fragments...

  1. Plant regeneration from proroplasts of alfalfa (Medicago sativa via somatic embryogenesis Regeneração de plantas a partir de protoplastos de alfafa (Medicago sativa via embriogênese somática

    Directory of Open Access Journals (Sweden)

    Mariza Monteiro

    2003-12-01

    Full Text Available Alfalfa is one of the most frequently studied species from the production of tissue culture-derived embryos point of view. In this study, five alfalfa cultivars were analyzed with reference to their ability to regenerate plants from protoplast cultures via somatic embryogenesis. Plant regeneration from leaf-derived protoplasts isolated from the cultivar Rangelander was achieved using a protocol defined for alfalfa cell suspension-derived embryogenesis. Because of its high efficiency, this procedure is recommended for protoplast electroporation-mediated genetic transformation of alfalfa.A alfafa é uma das espécies mais freqüentemente estudadas do ponto de vista da produção de embriões somáticos derivados da cultura de tecidos. Neste trabalho, cinco cultivares de alfafa foram analisados com referência à capacidade de regenerar plantas a partir de culturas de protoplastos via embriogênese somática. Regeneração de plantas a partir de protoplastos isolados de folhas da cultivar Rangelander foi obtida usando-se um protocolo definido para embriogênese somática derivada de suspensões celulares de alfafa. Em função da sua alta eficiência, recomenda-se o uso deste procedimento para transformação genética de alfafa mediada por eletroporação de protoplastos.

  2. In vitro regeneration in Allium species.

    Science.gov (United States)

    Rauber, M; Grunewaldt, J

    1988-10-01

    An attempt to induce shoot regeneration from leaf disc explants from Allium sativum L., A. porrum L., and A. schoenoprasum L. and the induction of shoot regeneration from single flower-bud receptacles in A. porrum is presented. While the regeneration rate from leaf disc explants was low, an efficient method for propagating A. porrum in vitro was obtained by cultivating single flower-bud receptacles. The shoot regeneration ability was strongly controlled by the genotype. Up to 294 shoots per leek plant could be harvested. Simultaneously the same plant could be used for seed production and bulbil formation in vivo. The efficiency of the in vitro multiplication method described allows the integration of this procedure into breeding programmes of A. porrum.

  3. Terrestrial Plant Biomarkers Preserved in Cariaco Basin Sediments: Records of Abrupt Tropical Vegetation Response to Rapid Climate Changes

    Science.gov (United States)

    Hughen, K. A.; Eglinton, T. I.; Makou, M.; Xu, L.; Sylva, S.

    2004-12-01

    Organic-rich sediments from the anoxic Cariaco Basin, Venezuela, preserve high concentrations of biomarkers for reconstruction of terrestrial environmental conditions. Molecular-level investigations of organic compounds provide a valuable tool for extracting terrestrial signals from these annually laminated marine sediments. Differences in hydrogen isotopic fractionation between C16-18 and C24-30 n-alkanoic acids suggest a marine source for the shorter chain lengths and a terrestrial source for the longer chains. Records of carbon and hydrogen isotopes, as well as average carbon chain length (ACL), from long-chain n-alkanoic acids parallel millennial-scale changes in vegetation and climate between the late Glacial and Preboreal periods, 15,000 to 10,000 years ago. Data from all terrestrial chain lengths were combined to produce single δ D and δ 13C indices through deglaciation, exhibiting enrichment during the late Glacial and Younger Dryas and depletion during the Bolling-Allerod and Preboreal periods. δ D reflects the hydrogen isotopic composition of environmental water used for plant growth, combined with evaporative enrichment within leaf spaces, and as such may act as a proxy for local aridity. Leaf wax δ 13C, which is a proxy for C3 versus C4 metabolic pathways, indicates that C3 plants predominated in the Cariaco watershed during warm/wet Bolling-Allerod and Holocene periods, and C4 plant biomass proliferated during cool/dry Glacial and Younger Dryas intervals. Coupled carbon and hydrogen isotopic measurements together clearly distinguish deglacial climatic periods as wetter with C3 vegetation versus drier with C4 vegetation. High resolution biomarker records reveal the rapidity of vegetation changes in northern South America during the last deglaciation. The leaf wax data reveal that local vegetation biomass, although not necessarily entire assemblages, shifted between arid grassland and wetter forest taxa on timescales of decades. Comparison of ACL

  4. POTENTIAL OF UAV BASED CONVERGENT PHOTOGRAMMETRY IN MONITORING REGENERATION STANDARDS

    Directory of Open Access Journals (Sweden)

    U. Vepakomma

    2015-08-01

    Full Text Available Several thousand hectares of forest blocks are regenerating after harvest in Canada. Monitoring their performance over different stages of growth is critical in ensuring future productivity and ecological balance. Tools for rapid evaluation can support timely and reliable planning of interventions. Conventional ground surveys or visual image assessments are either time intensive or inaccurate, while alternate operational remote sensing tools are unavailable. In this study, we test the feasibility and strength of UAV-based photogrammetry with an EO camera on a UAV platform in assessing regeneration performance. Specifically we evaluated stocking, spatial density and height distribution of naturally growing (irregularly spaced stems or planted (regularly spaced stems conifer regeneration in different phases of growth. Standard photogrammetric workflow was applied on the 785 acquired images for 3D reconstruction of the study sites. The required parameters were derived based on automated single stem detection algorithm developed in-house. Comparing with field survey data, preliminary results hold promise. Future studies are planned to expand the scope to larger areas and different stand conditions.

  5. Residual effect of growth regulators in etiolation and regeneration of in vitro pineapple plants Efeito residual de reguladores de crescimento no estiolamento e regeneração de plantas de abacaxi in vitro

    Directory of Open Access Journals (Sweden)

    Fernanda Vidigal Duarte Souza

    2010-06-01

    Full Text Available This work aimed to evaluate the influence of naphthaleneacetic acid (NAA and gibberellic acid (GA3 plant regulators in in vitro etiolation and subsequent regeneration of the PE x SC-60 pineapple hybrid. Nodal segments of in vitro plants with approximately 5-7 cm height were incubated in basic MS culture medium supplemented with 0.0; 0.5 and 1.0 mg L-1 of naphthaleneacetic acid (NAA in combination with gibberellic acid (GA3 in concentrations of 0.0; 0.5 and 1.0 mg L-1, and maintained at 27 ºC under dark condition. Evaluations were carried out at 90 and 180 days after incubation period. The best results for length of etiolated stems were obtained with 1.0 mg L-1 of NAA. In the experiment followed by the regeneration, stems with 3 cm from the etiolation treatment, were cultivated in proliferation medium and the number of regenerated plants per treatment was evaluated at 60 days of cultivation. The treatment that promoted the best etiolation of plants also promoted the worst regeneration rates, demonstrating the residual effect of the auxin used in the previous step in the regeneration of plants of the pineapple hybrid evaluated.O objetivo deste trabalho foi avaliar a influência dos reguladores vegetais ácido naftalenoacético (ANA e ácido giberélico (AG3 no estiolamento in vitro e posterior regeneração de plantas do abacaxizeiro híbrido PE x SC-60. Segmentos nodais de plantas produzidas in vitro, com aproximadamente 5-7 cm de comprimento, foram incubados em meio de cultura MS em sua concentração básica, suplementados com 0,0; 0,5 e 1,0 mg L-1 de ácido naftalenoacético (ANA em combinação com ácido giberélico (AG3, nas concentrações de 0,0; 0,5 e 1,0 mg L-1, e mantidos a 27ºC e sob condições de escuro. As avaliações foram realizadas aos 90; 180 e 270 dias de incubação. Os melhores resultados foram obtidos com a concentração de 1,0 mg L-1 de ANA. No experimento posterior de regeneração, hastes com 3 cm, oriundas dos

  6. DNA-based genetic markers for Rapid Cycling Brassica rapa (Fast Plants type designed for the teaching laboratory.

    Directory of Open Access Journals (Sweden)

    Eryn E. Slankster

    2012-06-01

    Full Text Available We have developed DNA-based genetic markers for rapid-cycling Brassica rapa (RCBr, also known as Fast Plants. Although markers for Brassica rapa already exist, ours were intentionally designed for use in a teaching laboratory environment. The qualities we selected for were robust amplification in PCR, polymorphism in RCBr strains, and alleles that can be easily resolved in simple agarose slab gels. We have developed two single nucleotide polymorphism (SNP based markers and 14 variable number tandem repeat (VNTR-type markers spread over four chromosomes. The DNA sequences of these markers represent variation in a wide range of genomic features. Among the VNTR-type markers, there are examples of variation in a nongenic region, variation within an intron, and variation in the coding sequence of a gene. Among the SNP-based markers there are examples of polymorphism in intronic DNA and synonymous substitution in a coding sequence. Thus these markers can serve laboratory exercises in both transmission genetics and molecular biology.

  7. Rapid ammonia gas transport accounts for futile transmembrane cycling under NH3/NH4+ toxicity in plant roots.

    Science.gov (United States)

    Coskun, Devrim; Britto, Dev T; Li, Mingyuan; Becker, Alexander; Kronzucker, Herbert J

    2013-12-01

    Futile transmembrane NH3/NH4(+) cycling in plant root cells, characterized by extremely rapid fluxes and high efflux to influx ratios, has been successfully linked to NH3/NH4(+) toxicity. Surprisingly, the fundamental question of which species of the conjugate pair (NH3 or NH4(+)) participates in such fluxes is unresolved. Using flux analyses with the short-lived radioisotope (13)N and electrophysiological, respiratory, and histochemical measurements, we show that futile cycling in roots of barley (Hordeum vulgare) seedlings is predominately of the gaseous NH3 species, rather than the NH4(+) ion. Influx of (13)NH3/(13)NH4(+), which exceeded 200 µmol g(-1) h(-1), was not commensurate with membrane depolarization or increases in root respiration, suggesting electroneutral NH3 transport. Influx followed Michaelis-Menten kinetics for NH3 (but not NH4(+)), as a function of external concentration (Km = 152 µm, Vmax = 205 µmol g(-1) h(-1)). Efflux of (13)NH3/(13)NH4(+) responded with a nearly identical Km. Pharmacological characterization of influx and efflux suggests mediation by aquaporins. Our study fundamentally revises the futile-cycling model by demonstrating that NH3 is the major permeating species across both plasmalemma and tonoplast of root cells under toxicity conditions.

  8. DNA-Based Genetic Markers for Rapid Cycling Brassica Rapa (Fast Plants Type) Designed for the Teaching Laboratory

    Science.gov (United States)

    Slankster, Eryn E.; Chase, Jillian M.; Jones, Lauren A.; Wendell, Douglas L.

    2012-01-01

    We have developed DNA-based genetic markers for rapid cycling Brassica rapa (RCBr), also known as Fast Plants. Although markers for B. rapa already exist, ours were intentionally designed for use in a teaching laboratory environment. The qualities we selected for were robust amplification in PCR, polymorphism in RCBr strains, and alleles that can be easily resolved in simple agarose slab gels. We have developed two single nucleotide polymorphism (SNP) based markers and 14 variable number tandem repeat (VNTR)-type markers spread over four chromosomes. The DNA sequences of these markers represent variation in a wide range of genomic features. Among the VNTR-type markers, there are examples of variation in a non-genic region, variation within an intron, and variation in the coding sequence of a gene. Among the SNP-based markers there are examples of polymorphism in intronic DNA and synonymous substitution in a coding sequence. Thus these markers can serve laboratory exercises in both transmission genetics and molecular biology. PMID:22675329

  9. Rapid detection by direct analysis in real time-mass spectrometry (DART-MS) of psychoactive plant drugs of abuse: the case of Mitragyna speciosa aka "Kratom".

    Science.gov (United States)

    Lesiak, Ashton D; Cody, Robert B; Dane, A John; Musah, Rabi A

    2014-09-01

    Mitragyna speciosa, also known commonly as "Kratom" or "Ketum", is a plant with psychoactive properties that have been attributed to the presence of various indole alkaloids such as mitragynine and 7-hydroxymitragynine. M. speciosa use is gaining popularity internationally as a natural and legal alternative to narcotics. As a drug of abuse, its detection and identification are not straightforward, since M. speciosa plant material is not particularly distinctive. Here, we show that direct analysis in real time-mass spectrometry (DART-MS) can be used not only to rapidly identify M. speciosa plant material and distinguish it from other plants, but also to distinguish between M. speciosa plant varieties, based on differences between their chemical profiles. The method is rapid and the analysis expeditious. Plant material such as that found at a crime scene can be analyzed directly with no sample pre-preparation steps. Furthermore, we show that the basis set of principal components that permit characterization of the plant material can be used to positively identify M. speciosa. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  10. Development of a triple hyphenated HPLC-radical scavenging detection-DAD-SPE-NMR system for the rapid identification of antioxidants in complex plant extracts

    NARCIS (Netherlands)

    Pukalskas, A.; Beek, van T.A.; Waard, de P.

    2005-01-01

    A rapid method for the simultaneous detection and identification of radical scavenging compounds in plant extracts was developed by combining an HPLC with on-line radical scavenging using DPPH as a model radical and an HPLC¿DAD¿SPE¿NMR system. Using this method a commercial rosemary extract was

  11. Development of a rapid and sensitive battery of bioassays for risk assessment of cyanobacterial microcystin-LR in drinking water of rural water treatment plants, South Africa

    CSIR Research Space (South Africa)

    Oberholster, Paul J

    2009-09-01

    Full Text Available become necessary to assess and develop additional methods that are rapid, yet realistic and cheap for the detection of cyanobacterial toxins in drinking water of rural conventional water treatment plants in South Africa. A well-known cyanobacterial...

  12. A rapid and robust method for simultaneously measuring changes in the phytohormones ABA, JA and SA in plants following biotic and abiotic stress

    Directory of Open Access Journals (Sweden)

    Mansfield John W

    2008-06-01

    Full Text Available Abstract We describe an efficient method for the rapid quantitative determination of the abundance of three acidic plant hormones from a single crude extract directly by LC/MS/MS. The method exploits the sensitivity of MS and uses multiple reaction monitoring and isotopically labelled samples to quantify the phytohormones abscisic acid, jasmonic acid and salicylic acid in Arabidopsis leaf tissue.

  13. A rapid and robust method for simultaneously measuring changes in the phytohormones ABA, JA and SA in plants following biotic and abiotic stress.

    OpenAIRE

    Forcat, S; Bennett, M; Mansfield, J; Grant, M

    2008-01-01

    Abstract We describe an efficient method for the rapid quantitative determination of the abundance of three acidic plant hormones from a single crude extract directly by LC/MS/MS. The method exploits the sensitivity of MS and uses multiple reaction monitoring and isotopically labelled samples to quantify the phytohormones abscisic acid, jasmonic acid and salicylic acid in Arabidopsis leaf tissue.

  14. Teaching Human Genetics with Mustard: Rapid Cycling "Brassica rapa" (Fast Plants Type) as a Model for Human Genetics in the Classroom Laboratory

    Science.gov (United States)

    Wendell, Douglas L.; Pickard, Dawn

    2007-01-01

    We have developed experiments and materials to model human genetics using rapid cycling "Brassica rapa", also known as Fast Plants. Because of their self-incompatibility for pollination and the genetic diversity within strains, "B. rapa" can serve as a relevant model for human genetics in teaching laboratory experiments. The experiment presented…

  15. An improved protocol for efficient transformation and regeneration of diverse indica rice cultivars

    Directory of Open Access Journals (Sweden)

    Sahoo Khirod K

    2011-12-01

    Full Text Available Abstract Background Rice genome sequencing projects have generated remarkable amount of information about genes and genome architecture having tremendous potential to be utilized in both basic and applied research. Success in transgenics is paving the way for preparing a road map of functional genomics which is expected to correlate action of a gene to a trait in cellular and organismal context. However, the lack of a simple and efficient method for transformation and regeneration is a major constraint for such studies in this important cereal crop. Results In the present study, we have developed an easy, rapid and highly efficient transformation and regeneration protocol using mature seeds as explants and found its successful applicability to a choice of elite indica rice genotypes. We have optimized various steps of transformation and standardized different components of the regeneration medium including growth hormones and the gelling agent. The modified regeneration medium triggers production of large number of shoots from smaller number of calli and promotes their faster growth, hence significantly advantageous over the existing protocols where the regeneration step requires maximum time. Using this protocol, significantly higher transformation efficiency (up to 46% and regeneration frequency (up to 92% for the untransformed calli and 59% for the transformed calli were achieved for the four tested cultivars. We have used this protocol to produce hundreds of independent transgenic lines of different indica rice genotypes. Upon maturity, these transgenic lines were fertile thereby indicating that faster regeneration during tissue culture did not affect their reproductive potential. Conclusions This speedy, yet less labor-intensive, protocol overcomes major limitations associated with genetic manipulation in rice. Moreover, our protocol uses mature seeds as the explant, which can easily be obtained in quantity throughout the year and kept

  16. Thidiazuron-induced regeneration of Echinacea purpurea L.: micropropagation in solid and liquid culture systems.

    Science.gov (United States)

    Jones, Maxwell P A; Yi, Zhijun; Murch, Susan J; Saxena, Praveen K

    2007-01-01

    The goals of this study were to investigate thidiazuron (TDZ)-induced morphogenesis of Echinacea purpurea L. and to assess the possibility of developing a liquid-based protocol for rapid micropropagation. Callus development and root organogenesis were observed on leaf explants cultured on media containing 2,4-dicholorophenoxyacetic acid or dicamba, but no plantlets were regenerated. Addition of TDZ to the culture medium as the sole growth regulator resulted in the production of regenerable callus cultures. The highest rate of regeneration was observed for explants cultured on medium with TDZ at 2.5 microM or higher. Tissue derived from 1.0 microM TDZ treatments was used to initiate liquid cultures. All liquid treatments produced a similar number of regenerants but significantly more healthy plants were obtained from cultures grown in the presence of 0.1 and 1.0 microM TDZ. This TDZ-based micropropagation system is the first liquid, large-scale propagation protocol developed for the mass production of E. purpurea plants.

  17. Injuries on Seedlings Caused by Potential Weed in Tropical Rain Forest Regeneration Areas

    Directory of Open Access Journals (Sweden)

    Sumardi Sumardi

    2000-07-01

    Full Text Available The experiment aimed to assess the injuries on Shorea seedlings caused by weed in artificial regeneration of tropical rain forest in Jambi. Four planting systems, strip nurse planting (using Acacia mangium, Paraserianthes falcataria and Gmelina arborea, line planting, gap planting and natural regeneration, were used. Seedling injuries were assessed based on part of seedling suppressed. Results indicated that Shorea seedling suffered from varying degrees of injuries, depending on weed species and part of the seedling suppressed. The dominance of weed and damage intensity were determined by the level of canopy opening on the planting systems. Ground cover dominated rapidly in open canopy, causing up to 55.27% injuries on the seedlings in the strips of G. arborea and P. falcataria. Whereas creepers and vines became dominant in moderate canopy opening. The injury of Shorea seedling planted under nurse tree was determined by the species and planting density of nurse tree used. Light canopy nurse tree such as P. falcataria failed to suppress ground weed, but in the contrary, A. mangium with heavy canopy still allowed creepers and vines to grow.

  18. Helping the Retina Regenerate

    Science.gov (United States)

    ... report summarized two possible therapeutic strategies for RGC regeneration. The first would use stem cells to grow ... recommendations in the report include systematic comparisons of animal models that do and do not regenerate RGCs, ...

  19. Regeneration of plantlets under NaCl stress from NaN3 treated ...

    African Journals Online (AJOL)

    Maximum of 8.41 ± 0.36 plantlets callus-1 were regenerated in MS4 (control) culture of Thatta-10, and 4.94 ± 0.05 plantlets of CPF-237 in 25 mol m-3 NaCl stressed plant regeneration (MS4a) medium. Plant regeneration on MS4b (2.21 ± 0.17 plantlets callus-1) was observed in CPF-237 only. Regenerated plantlets were ...

  20. Subalpine vegetation pattern three decades after stand-replacing fire: Effects of landscape context and topography on plant community composition, tree regeneration, and diversity

    Science.gov (United States)

    Jonathan D. Coop; Robert T. Massatti; Anna W. Schoettle

    2010-01-01

    These subalpine wildfires generated considerable, persistent increases in plant species richness at local and landscape scales, and a diversity of plant communities. The findings suggest that fire suppression in such systems must lead to reduced diversity. Concerns about post-fire invasion by exotic plants appear unwarranted in high-elevation wilderness settings.

  1. A Protocol for Rapid, Measurable Plant Tissue Culture Using Stem Disc Meristem Micropropagation of Garlic ("Allium Sativum L.")

    Science.gov (United States)

    Peat, Gerry; Jones, Meriel

    2012-01-01

    Plant tissue culture is becoming an important technique for the mass propagation of plants. Problems with existing techniques, such as slow growth and contamination, have restricted the practical work in plant tissue culture carried out in schools. The new protocol using garlic meristematic stem discs explained in this article addresses many of…

  2. An efficient callus initiation and direct regeneration of Stevia ...

    African Journals Online (AJOL)

    The experiment was carried out to establish an efficient callus induction system of Stevia rebaudiana from a variety of explants as well as direct regeneration from nodes. Callus initiation was done to extract secondary metabolites. However direct regeneration is essential for rapid large-scale clonal propagation. Among leaf ...

  3. Highly efficient in vitro adventitious shoot regeneration of Adenosma ...

    African Journals Online (AJOL)

    Yomi

    2012-04-10

    Apr 10, 2012 ... perfume plant Adenosma buchneroides Bonati. In: Proceed. Seventh. Perfume Plant Congress, pp. 26-29. Zhou HC, Li M, Zhao X, Fan XC, Guo AG (2010). Plant regeneration from in vitro leaves of the peach rootstock 'Nemaguard' (Prunus persica × P. davidiana). Plant Cell, Tiss. Org. Cult. 101: 79-87.

  4. MHD (Magnetohydrodynamics) recovery and regeneration

    Energy Technology Data Exchange (ETDEWEB)

    McIlroy, R. A. [Babcock and Wilcox Co., Alliance, OH (United States). Research Center; Probert, P. B. [Babcock and Wilcox Co., Alliance, OH (United States). Research Center; Lahoda, E. J. [Westinghouse Electric Corp., Pittsburgh, PA (United States); Swift, W. M. [Argonne National Lab. (ANL), Argonne, IL (United States); Jackson, D. M. [Univ. of Tennessee Space Inst. (UTSI), Tullahoma, TN (United States); Prasad, J. [Univ. of Tennessee Space Inst. (UTSI), Tullahoma, TN (United States); Martin, J. [Hudson Engineering (United States); Rogers, C. [Hudson Engineering (United States); Ho, K. K. [Babcock and Wilcox Co., Alliance, OH (United States). Research Center; Senary, M. K. [Babcock and Wilcox Co., Alliance, OH (United States). Research Center; Lee, S. [Univ. of Akron, OH (United States)

    1988-10-01

    A two-phase program investigating MHD seed regeneration is described. In Phase I, bench scale experiments were carried out to demonstrate the technical feasibility of a proposed Seed Regeneration Process. The Phase I data has been used for the preliminary design of a Proof-of-Concept (POC) plant which will be built and tested in Phase II. The Phase I data will also be used to estimate the costs of a 300 Mw(t) demonstration plant for comparison with other processes. The Seed Regeneration Process consists of two major subprocesses; a Westinghouse Dry Reduction process and a modified Tampella (sulfur) Recovery process. The Westinghouse process reduces the recovered spent seed (i.e., potassium sulfate) to potassium polysulfide in a rotary kiln. The reduction product is dissolved in water to form green liquor, clarified to remove residual coal ash, and sent to the Tampella sulfur release system. The sulfur is released using carbon dioxide from flue gas in a two stage reaction. The sulfur is converted to elemental sulfur as a marketable by product. The potassium is crystallized from the green liquor and dried to the anhydrous form for return to the MHD unit.

  5. Regeneration of periodontal tissues: guided tissue regeneration.

    Science.gov (United States)

    Villar, Cristina C; Cochran, David L

    2010-01-01

    The concept that only fibroblasts from the periodontal ligament or undifferentiated mesenchymal cells have the potential to re-create the original periodontal attachment has been long recognized. Based on this concept, guided tissue regeneration has been applied with variable success to regenerate periodontal defects. Quantitative analysis of clinical outcomes after guided tissue regeneration suggests that this therapy is a successful and predictable procedure to treat narrow intrabony defects and class II mandibular furcations, but offers limited benefits in the treatment of other types of periodontal defects.

  6. Mechanisms of lymphatic regeneration after tissue transfer.

    Directory of Open Access Journals (Sweden)

    Alan Yan

    2011-02-01

    Full Text Available Lymphedema is the chronic swelling of an extremity that occurs commonly after lymph node resection for cancer treatment. Recent studies have demonstrated that transfer of healthy tissues can be used as a means of bypassing damaged lymphatics and ameliorating lymphedema. The purpose of these studies was to investigate the mechanisms that regulate lymphatic regeneration after tissue transfer.Nude mice (recipients underwent 2-mm tail skin excisions that were either left open or repaired with full-thickness skin grafts harvested from donor transgenic mice that expressed green fluorescent protein in all tissues or from LYVE-1 knockout mice. Lymphatic regeneration, expression of VEGF-C, macrophage infiltration, and potential for skin grafting to bypass damaged lymphatics were assessed.Skin grafts healed rapidly and restored lymphatic flow. Lymphatic regeneration occurred beginning at the peripheral edges of the graft, primarily from ingrowth of new lymphatic vessels originating from the recipient mouse. In addition, donor lymphatic vessels appeared to spontaneously re-anastomose with recipient vessels. Patterns of VEGF-C expression and macrophage infiltration were temporally and spatially associated with lymphatic regeneration. When compared to mice treated with excision only, there was a 4-fold decrease in tail volumes, 2.5-fold increase in lymphatic transport by lymphoscintigraphy, 40% decrease in dermal thickness, and 54% decrease in scar index in skin-grafted animals, indicating that tissue transfer could bypass damaged lymphatics and promote rapid lymphatic regeneration.Our studies suggest that lymphatic regeneration after tissue transfer occurs by ingrowth of lymphatic vessels and spontaneous re-connection of existing lymphatics. This process is temporally and spatially associated with VEGF-C expression and macrophage infiltration. Finally, tissue transfer can be used to bypass damaged lymphatics and promote rapid lymphatic regeneration.

  7. Accelerated cell divisions drive the outgrowth of the regenerating spinal cord in axolotls

    National Research Council Canada - National Science Library

    Rost, Fabian; Rodrigo Albors, Aida; Mazurov, Vladimir; Brusch, Lutz; Deutsch, Andreas; Tanaka, Elly M; Chara, Osvaldo

    2016-01-01

    .... Previously, we showed that regenerating stem cells in the axolotl spinal cord revert to a molecular state resembling embryonic neuroepithelial cells and functionally acquire rapid proliferative divisions...

  8. Rapid Transient Production in Plants by Replicating and Non-Replicating Vectors Yields High Quality Functional Anti-HIV Antibody

    OpenAIRE

    Frank Sainsbury; Markus Sack; Johannes Stadlmann; Heribert Quendler; Rainer Fischer; Lomonossoff, George P.

    2010-01-01

    Background The capacity of plants and plant cells to produce large amounts of recombinant protein has been well established. Due to advantages in terms of speed and yield, attention has recently turned towards the use of transient expression systems, including viral vectors, to produce proteins of pharmaceutical interest in plants. However, the effects of such high level expression from viral vectors and concomitant effects on host cells may affect the quality of the recombinant product. Meth...

  9. A reproducible protocol for regeneration and transformation in ...

    African Journals Online (AJOL)

    The objective of the present study is to develop an efficient protocol for shoot and plant regeneration using five commercial canola cultivars grown under the Egyptian agricultural conditions. The regeneration efficiency from hypocotyl explants was examined. The data indicated that embryonic calli were formed within two ...

  10. Highly efficient in vitro adventitious shoot regeneration of Adenosma ...

    African Journals Online (AJOL)

    Adenosma glutinosum (Linn.) Druce is an important aromatic plant, but no information is available regarding its regeneration, callus induction and proliferation from leaf explants. In this study, an in vitro shoot regeneration procedure was developed for native A. glutinosum using leaf explants. Callus induction and shoots ...

  11. Reproducible in vitro regeneration system for purifying sugarcane ...

    African Journals Online (AJOL)

    Genome purification of a selected clone of sugarcane is the key to developing homogenous lines. Generally, regenerated plants after transformation are heterogeneous at genome level, and several successive rounds of selection on antibiotic-containing medium and regeneration cycles are required to purify the genome to ...

  12. In vitro regeneration, detection of somaclonal variation and ...

    African Journals Online (AJOL)

    Enzyme linked immunosorbent assay (ELISA) test was performed to detect the presence of sugarcane mosaic virus (SCMV) in the regenerated plantlets and simple sequence repeat (SSR) markers were used to evaluate the genetic variation at DNA level between the parent's plants and regenerated somaclones of the ...

  13. Germination and In Vitro Regeneration Response of Local Nigerian ...

    African Journals Online (AJOL)

    Prof. Ogunji

    Abstract. In vitro regeneration of plants is an important tool in both basic and applied studies as well as commercial applications. It can be of use in overcoming the constraints of tomato production. Assessment of seed sterilization procedures and in vitro regeneration of tomato cultivar was attempted using different explant ...

  14. Germination and In Vitro regeneration response of local Nigerian ...

    African Journals Online (AJOL)

    In vitro regeneration of plants is an important tool in both basic and applied studies as well as commercial applications. It can be of use in overcoming the constraints of tomato production. Assessment of seed sterilization procedures and in vitro regeneration of tomato cultivar was attempted using different explant sources.

  15. Embriogênese somática e regeneração de plantas a partir de embrião maduro de aveia Somatic embryogenesis and plant regeneration derived from mature embryos of oat

    Directory of Open Access Journals (Sweden)

    Caren Regina Cavichioli Lamb

    2002-02-01

    Full Text Available Calo embriogênico tem sido o tecido-alvo mais utilizado para transformação genética de cereais. O objetivo deste trabalho foi investigar o estabelecimento de calos embriogênicos e a regeneração de plantas in vitro a partir de embriões maduros de genótipos de aveia (Avena sativa L.. Embriões maduros foram retirados das sementes e colocados em meio MS (Murashige & Skoog, contendo 30,0 g L-1 de sacarose e 2,0 mg L-1 de ácido 2,4-diclorofenoxiacético (2,4-D. Após o período de indução de calos, agregados embriogênicos foram isolados e subcultivados a cada 21 dias para meio fresco. Os calos embriogênicos foram então transferidos para meio de indução de parte aérea, e, na seqüência, as partes aéreas foram transferidas para meio de indução de raízes. Houve diferenças entre genótipos quanto à capacidade de embriogênese somática e regeneração de plantas in vitro a partir de embrião maduro. Este explante permitiu a indução de calos embriogênicos, que se multiplicaram, e que regeneraram in vitro um grande número de plantas de genótipos como UFRGS 7 e UFRGS 19, o que o faz passível de ser utilizado na transformação genética da aveia.Embryogenic callus has been the most used target tissue for cereal genetic transformation. Therefore, the objective of this study was to investigate the establishment of embryogenic calli and the in vitro plant regeneration from mature embryos of oat genotypes (Avena sativa L.. Mature embryos were taken out of the seeds and placed on a culture medium MS (Murashige & Skoog, containing 30,0 mg L-1 of sucrose and 2,0 mg L-1 of 2,4-dichlorophenoxyacetic acid (2,4-D. From the induction period, embryogenic aggregates were isolated and subcultivated each 21 days into a fresh medium. After this period, embryogenic calli were transferred to a medium for shoot regeneration. Subsequently, the shoot was transferred to a medium for root induction. There was variability among genotypes for somatic

  16. A Novel System for Rapid and Cost-Effective Production of Detection and Diagnostic Reagents of West Nile Virus in Plants

    Directory of Open Access Journals (Sweden)

    Junyun He

    2012-01-01

    Full Text Available The threat of West Nile virus (WNV epidemics necessitates the development of a technology platform that can produce reagents to support detection and diagnosis rapidly and inexpensively. A plant expression system is attractive for protein production due to its low-cost and high-scalability nature and its ability to make appropriate posttranslational modifications. Here, we investigated the feasibility of using plants to produce two WNV detection and diagnostic reagents to address the current cost and scalability issues. We demonstrated that WNV DIII antigen and E16 monoclonal antibody are rapidly produced at high levels in two plant species and are easily purified. Furthermore, they are effective in identifying WNV and in detecting human IgM response to WNV infection. E16 mAb does not cross-react with other flaviviruses, therefore, is valuable for improving diagnostic accuracy. This study provides a proof of principle for using plants as a robust and economical system to produce diagnostic reagents for arboviruses.

  17. Rapid Analytical Method for the Determination of Aflatoxins in Plant-Derived Dietary Supplement and Cosmetic Oils

    Science.gov (United States)

    Consumption of edible oils derived from conventional crop plants is increasing because they are generally regarded as more healthy alternatives to animal based fats and oils. More recently there has been increased interest in the use of alternative specialty plant-derived oils, including those from...

  18. Plant lesions promote the rapid multiplication of Escherichia coli O157:H7 on post-harvest lettuce

    Science.gov (United States)

    Several outbreaks of Escherichia coli O157:H7 (EcO157) infections have been associated with minimally processed leafy vegetables in the U.S. Harvesting and processing cause plant tissue damage. In order to assess the role of plant tissue damage in the contamination of leafy greens with EcO157, the e...

  19. Desulfurization sorbent regeneration

    Science.gov (United States)

    Jalan, V.M.; Frost, D.G.

    1982-07-07

    A spent solid sorbent resulting from the removal of hydrogen sulfide from a fuel gas flow is regenerated with a steam-air mixture. The mixture of steam and air may also include additional nitrogen or carbon dioxide. The gas mixture contacts the spent sorbent containing metal sulfide at a temperature above 500/sup 0/C to regenerate the sulfide to metal oxide or carbonate. Various metal species including the period four transition metals and the lanthanides are suitable sorbents that may be regenerated by this method. In addition, the introduction of carbon dioxide gas permits carbonates such as those of strontium, barium and calcium to be regenerated. The steam permits regeneration of spent sorbent without formation of metal sulfate. Moreover, the regeneration will proceed with low oxygen concentrations and will occur without the increase in temperature to minimize the risk of sintering and densification of the sorbent. This method may be used for high-temperature fuel cells.

  20. Artificial regeneration of major oak (Quercus) species in the eastern United States - a review of the literature

    Science.gov (United States)

    Daniel C. Dey; Douglass Jacobs; Ken McNabb; Gary W. Miller; V. Baldwin; G. Foster

    2008-01-01

    Although natural regeneration is often the best method for establishing new oak (Quercus spp.) stands, there are increasingly more situations in which high potential for oak regeneration failure dictates the use of artificial regeneration including direct seeding and planting seedlings. Additionally, afforestation planting programs frequently...