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 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 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

scholarly journals Establishment of a Tractable Genetic Transformation System in Veillonella spp.

2012 ◽  
Vol 78 (9) ◽  
pp. 3488-3491 ◽  
Author(s):  
Jinman Liu ◽  
Zhoujie Xie ◽  
Justin Merritt ◽  
Fengxia Qi
Keyword(s):  
Escherichia Coli ◽  
Genetic Transformation ◽  
Shuttle Vector ◽  
Transformation System ◽  
Genetic Studies ◽  
Content Type ◽  
Genetic Transformation System

ABSTRACTWe have constructed the firstEscherichia coli-Veillonellashuttle vector based on an endogenous plasmid (pVJL1) isolated from a clinicalVeillonellastrain. A highly transformableVeillonellastrain was also identified. Both the shuttle vector and the transformable strain should be valuable tools for futureVeillonellagenetic studies.

scholarly journals Robust Genetic Transformation System to Obtain Non-chimeric Transgenic Chickpea

2019 ◽  
Vol 10 ◽  
Author(s):  
Sudipta Shekhar Das Bhowmik ◽  
Alam Yen Cheng ◽  
Hao Long ◽  
Grace Zi Hao Tan ◽  
Thi My Linh Hoang ◽  
...  
Keyword(s):  
Genetic Transformation ◽  
Transformation System ◽  
Genetic Transformation System

scholarly journals Construction of a Genetic Transformation System for a Freeze-tolerant YeastKluyveromyces thermotolerans

1992 ◽  
Vol 56 (2) ◽  
pp. 228-232 ◽  
Author(s):  
Akihiro Hino ◽  
Chakamas Wongkhalaung ◽  
Shinya Kawai ◽  
Satoru Murao ◽  
Keiji Yano ◽  
...  
Keyword(s):  
Genetic Transformation ◽  
Transformation System ◽  
Freeze Tolerant ◽  
Genetic Transformation System

scholarly journals Agrobacterium tumefaciens-Mediated Transformation for Investigation of Somatic Recombination in the Fungal Pathogen Armillaria mellea

2010 ◽  
Vol 76 (24) ◽  
pp. 7990-7996 ◽  
Author(s):  
Kendra Baumgartner ◽  
Phillip Fujiyoshi ◽  
Gary D. Foster ◽  
Andy M. Bailey
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Genetic Transformation ◽  
Natural Populations ◽  
Reproductive Mode ◽  
Mitochondrial Haplotype ◽  
Transformation System ◽  
Serial Transfer ◽  
Haploid Nucleus ◽  
Somatic Recombination ◽  
Genetic Transformation System

ABSTRACT Armillaria root disease is one of the most damaging timber and fruit tree diseases in the world. Despite its economic importance, many basic questions about the biology of the causal fungi, Armillaria spp., are unanswered. For example, Armillaria undergoes matings between diploid and haploid mycelia, which can result in a recombinant diploid without meiosis. Evidence of such somatic recombination in natural populations suggests that this reproductive mode may affect the pathogen's ecology. Investigations of the mechanisms and adaptive consequences of somatic recombination are, however, hampered by the lack of a method to reliably synthesize somatic recombinants. Here we report the first genetic transformation system for the genus Armillaria. We transformed A. mellea with selective markers for use in diploid-haploid matings to reliably synthesize somatic recombinants. This was accomplished with Agrobacterium tumefaciens carrying pBGgHg, which carries the hygromycin phosphotransferase gene (hph). hph was integrated into transformants, as evidenced by serial transfer to selective media, PCR, reverse transcription-PCR (RT-PCR), and Southern hybridization. Nuclear and mitochondrial markers were developed to genotype synthesized mycelia. In matings between a wild-type diploid and hygromycin-resistant haploids (transgenic), we identified recombinant, hygromycin-resistant diploids and, additionally, hygromycin-resistant triploids, all with the mitochondrial haplotype of the haploid partner. Our approach created no mycelium in which the haploid nucleus was replaced by the diploid nucleus, the typical outcome of diploid-haploid matings in Armillaria. This genetic transformation system, in combination with new markers to track chromosomal and cytoplasmic inheritance in A. mellea, will advance research aimed at characterizing the significance of somatic recombination in the ecology of this important fungus.

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