scholarly journals Standardized Genetic Transformation Protocol for Chrysanthemum cv. ‘Jinba’ with TERMINAL FLOWER 1 Homolog CmTFL1a

Genes ◽  
2020 ◽  
Vol 11 (8) ◽  
pp. 860
Author(s):  
Saba Haider ◽  
Yaohui Gao ◽  
Yike Gao
Keyword(s):  
Transgenic Plants ◽  
Genetic Transformation ◽  
Model Organism ◽  
Plant Morphology ◽  
Pcr Analysis ◽  
Transformation System ◽  
Transformation Protocol ◽  
Genetic Transformation System ◽  
Chrysanthemum X Morifolium

Chrysanthemum (Chrysanthemum x morifolium Ramat.) cultivar Jinba is a distinctive short-day chrysanthemum that can be exploited as a model organism for studying the molecular mechanism of flowering. The commercial value of Jinba can be increased in global flower markets by developing its proper regeneration and genetic transformation system. By addressing typical problems associated with Agrobacterium-mediated transformation in chrysanthemum, that is, low transformation efficiency and high cultivar specificity, we designed an efficient, stable transformation system. Here, we identify the features that significantly affect the genetic transformation of Jinba and standardize its transformation protocol by using CmTFL1a as a transgene. The appropriate concentrations of various antibiotics (kanamycin, meropenem and carbenicillin) and growth regulators (6-BA, 2,4-D and NAA) for the genetic transformation were determined to check their effects on in vitro plant regeneration from leaf segments of Jinba; thus, the transformation protocol was standardized through Agrobacterium tumefaciens (EHA105). In addition, the presence of the transgene and its stable expression in CmTFL1a transgenic plants were confirmed by polymerase chain reaction (PCR) analysis. The CmTFL1a transgene constitutively expressed in the transgenic plants was highly expressed in shoot apices as compared to stem and leaves. Overexpression of CmTFL1a led to a delay in transition to the reproductive phase and significantly affected plant morphology. This study will help to understand the biological phenomenon of TFL1 homolog in chrysanthemum. Moreover, our findings can explore innovative possibilities for genetic engineering and breeding of other chrysanthemum cultivars.

scholarly journals DEVELOPMENT OF THE DUCKWEED (LEMNA MINOR) GENETIC TRANSFORMATION SYSTEM

2008 ◽  
Vol 6 (4) ◽  
pp. 20-28 ◽  
Author(s):  
Sofia E Gaydukova ◽  
Andrey L Rakitin ◽  
Nikolay V Ravin ◽  
Konstantin G Skryabin ◽  
Anastasia M Kamionskaya
Keyword(s):  
Transgenic Plants ◽  
Genetic Transformation ◽  
Molecular Analysis ◽  
Recombinant Proteins ◽  
Water Body ◽  
Lemna Minor ◽  
Transformation System ◽  
Effective Selection ◽  
Genetic Transformation System

Regeneration parameters were optimized and effective selection conditions were sorted out. Transgenic plants of Lemna minor were obtained and their status was confirmed by molecular analysis. Optimized methodology can be used for obtaining transgenic duckweed plants producing recombinant proteins and water body bioremediators.

scholarly journals Establishment of a Plant Regeneration and Genetic Transformation System of Panax ginseng C.A. Meyer

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 572d-572
Author(s):  
Hak-Tae Lim ◽  
Haeng-Soon Lee ◽  
Tage Eriksson
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Genetic Transformation ◽  
Adventitious Shoots ◽  
Shoot Multiplication ◽  
Regeneration System ◽  
Transformation Protocol ◽  
Genetic Transformation System ◽  
Regeneration Systems

Plant regeneration of ginseng has been known to be difficult, and there are a few reports on plant regeneration of ginseng via somatic embryogenesis. In vitro flowering has, however, been one of the major drawbacks in these regeneration systems in which BA and GA3 were included in germination and shoot multiplication media. Multiplication of adventitious shoots from a single somatic embryo, abnormal morphology, and vitrified shoots were also observed. All these facts have made successful acclimatization of ginseng plantlets difficult. The purposes of this study were 1) to establish the plant regeneration system via organogenesis, 2) to improve normal plant regeneration via somatic embryogenesis, 3) to improve the efficiency of plant regeneration from protoplast culture, 4) to understand the acclimatization process, 5) to develop effective genetic transformation protocol. Data in relation with all these studies are presented in detail.

scholarly journals Genotype Independent Regeneration and Agrobacterium?mediated Genetic Transformation of Sweet Potato (Ipomoea batatas L.)

2013 ◽  
Vol 23 (1) ◽  
Author(s):  
Shubhendu Shekhar ◽  
Lalit Agrawal ◽  
Alak Kumar Buragohain ◽  
Asis Datta ◽  
Subhra Chakraborty ◽  
...  
Keyword(s):  
Genetic Transformation ◽  
Sweet Potato ◽  
Ipomoea Batatas ◽  
Shoot Formation ◽  
Pcr Analysis ◽  
Transcript Accumulation ◽  
Agrobacterium Strain ◽  
Transformation Protocol ◽  
Genetic Transformation System ◽  
Stem Segments

Development of an efficient genotype independent regeneration and genetic transformation system in sweet potato continues to be of great interest. Agrobacterium?mediated genetic transformation protocol was established in two different cultivars of sweet potato using Agrobacterium strain EHA105 harbouring binary plasmid pBI121 containing GUS and nptII genes. The internodal stem segments from 30?day?old micropropogated plants were used as explant with different combinations of media and hormones. MS and LS media with various concentrations of growth regulators proved to be non?responsive and the infecundity was severe with the addition of cytokinins. Nonetheless, MS with 2,4?D and TDZ gave a good percentage of callusing but with low differentiation. In different concentrations of NAA, significant amount of callusing was observed but percentage of rooting remained low in both the genotypes. Gamborg’s B5 supplemented with NAA proved to be the most suitable media and hormone combination, which yielded shoot formation after 8 ? 10 weeks with a regenera? tion efficiency of 40 ? 70%. Stable integration of transgene was confirmed by PCR analysis. Furthermore, qRT?PCR analysis was performed to assess the transcript accumulation in addition to the GUS enzymatic assay in the transgenic lines.Plant Tissue Cult. & Biotech. 23(1): 87?100, 2013 (June)DOI: http://dx.doi.org/10.3329/ptcb.v23i1.15565D.O.I. 10.3329/ptcb.v23i1.15565

scholarly journals Agrobacterium-mediated Genetic Transformation in Lentil (Lens culinaris Medik.) followed by In vitro Flowering and Seed Formation

2012 ◽  
Vol 22 (1) ◽  
pp. 13-26 ◽  
Author(s):  
Subroto K. Das ◽  
Kishwar Jahan Shethi ◽  
M. I. Hoque ◽  
R. H. Sarker
Keyword(s):  
Genetic Transformation ◽  
Lens Culinaris ◽  
Multiple Shoots ◽  
Cotyledonary Node ◽  
In Vitro Flowering ◽  
Pcr Analysis ◽  
Seed Formation ◽  
Half Strength ◽  
Genetic Transformation System

Genetic transformation system was developed for two microsperma varieties of lentil (Lens culinaris Medik.), namely Bari Masur-4 (BM-4) and Bari Masur-5 (BM-5) using Agrobacterium tumefaciens strain LBA4404 harbouring binary plasmid pBI121, containing GUS and nptII genes. Three different types of embryo explants, namely cotyledonary node (CN), decapitated embryo (DE) and cotyledone attached decapitated embryo (CADE) were used. Highest GUS positive expression was found in DE followed by CADE as detected by transient assays. Following Agrobacterium infection CADE showed better response in developing multiple shoots on MS supplemented with 2.22 µM BAP, 2.32 µM Kn, 0.29 µM GA3 and 30.35 µM tyrosine. Selection of the transformed shoots was carried out by gradually increasing the concentration of kanamycin up to 200 mg/l. Transgenic lentil shoots were produced with an overall frequency of 1.009%. In vitro rooting appeared to have a limitation in obtaining complete plantlets in lentil, therefore in vitro flowering and seed formation were induced in transformed shoots of lentil with a view to recovering of the transgenic progenies.  GUS positive shoots were found to produce in vitro flowers and pods on half-strength MS containing 98.4 µM IBA and 2.69 µM NAA. Expression of gene was detected in various tissues of the transformed shoots. Stable integration of GUS gene was also confirmed through PCR analysis. Plant Tissue Cult. & Biotech. 22(1): 13-26, 2012 (June) DOI: http://dx.doi.org/10.3329/ptcb.v22i1.11243 

scholarly journals Establishment of a Genetic Transformation System in Guanophilic Fungus Amphichorda guana

2021 ◽  
Vol 7 (2) ◽  
pp. 138
Author(s):  
Min Liang ◽  
Wei Li ◽  
Landa Qi ◽  
Guocan Chen ◽  
Lei Cai ◽  
...  
Keyword(s):  
Genetic Transformation ◽  
Genetic Manipulation ◽  
Major Facilitator Superfamily ◽  
Protoplast Regeneration ◽  
Transformation System ◽  
Regeneration Rate ◽  
Bioactive Natural Products ◽  
Genetics Research ◽  
Genetic Transformation System ◽  
Major Facilitator

Fungi from unique environments exhibit special physiological characters and plenty of bioactive natural products. However, the recalcitrant genetics or poor transformation efficiencies prevent scientists from systematically studying molecular biological mechanisms and exploiting their metabolites. In this study, we targeted a guanophilic fungus Amphichorda guana LC5815 and developed a genetic transformation system. We firstly established an efficient protoplast preparing method by conditional optimization of sporulation and protoplast regeneration. The regeneration rate of the protoplast is up to about 34.6% with 0.8 M sucrose as the osmotic pressure stabilizer. To develop the genetic transformation, we used the polyethylene glycol-mediated protoplast transformation, and the testing gene AG04914 encoding a major facilitator superfamily transporter was deleted in strain LC5815, which proves the feasibility of this genetic manipulation system. Furthermore, a uridine/uracil auxotrophic strain was created by using a positive screening protocol with 5-fluoroorotic acid as a selective reagent. Finally, the genetic transformation system was successfully established in the guanophilic fungus strain LC5815, which lays the foundation for the molecular genetics research and will facilitate the exploitation of bioactive secondary metabolites in fungi.

Establishment of efficient callus genetic transformation system for Pyrus armeniacaefolia

2021 ◽  
Vol 289 ◽  
pp. 110429
Author(s):  
Xinhui Wang ◽  
Fengli Zhou ◽  
Jianlong Liu ◽  
Wenqian Liu ◽  
Shaoling Zhang ◽  
...  
Keyword(s):  
Genetic Transformation ◽  
Transformation System ◽  
Genetic Transformation System
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