Genetic transformation system for Bacillus velezensis NSZ-YBGJ001 and curing of the endogenous plasmid pBV01

2021 ◽  
Author(s):  
Hui Tian ◽  
Bo Liu ◽  
Juan Yang ◽  
Chengfang Zhou ◽  
Xinxin Xu ◽  
...  
Keyword(s):  
Genetic Transformation ◽  
Transformation System ◽  
Bacillus Velezensis ◽  
Genetic Transformation System

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.

Efficient plant regeneration and genetic transformation system of the precious fast-growing tree Toona ciliata

2021 ◽  
Vol 172 ◽  
pp. 114015
Author(s):  
Wenmai Mao ◽  
Huiyun Song ◽  
Yue Li ◽  
Yueyang Wang ◽  
Huijuan Lin ◽  
...  
Keyword(s):  
Plant Regeneration ◽  
Genetic Transformation ◽  
Transformation System ◽  
Fast Growing ◽  
Toona Ciliata ◽  
Genetic Transformation System

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.

Establishment of regeneration system and transformation of Zm401 gene in Lilium longiflorum×L. formosanum

2008 ◽  
Vol 5 (2) ◽  
pp. 113-119 ◽  
Author(s):  
Li Qiu-Hua ◽  
Hong Bo ◽  
Tong Zheng ◽  
Ma Chao ◽  
Guan Ai-Nong ◽  
...  
Keyword(s):  
Genetic Transformation ◽  
Lilium Longiflorum ◽  
Bacterial Suspension ◽  
Specific Gene ◽  
Naphthaleneacetic Acid ◽  
Transformation System ◽  
Regeneration System ◽  
Regeneration Rate ◽  
Maize Pollen ◽  
Genetic Transformation System

AbstractIn vitro bulb scales of Lilium longiflorum×L. formosanum were used as explants to develop a highly efficient regeneration system. A high regeneration rate (100%) was reached through organogenesis on basal Murashige and Skoog (MS) medium supplemented with 1.0 mg/l 6-benzylaminopurine (6-BA) and 1.0 mg/l naphthaleneacetic acid (NAA). A genetic transformation system for the lily was developed using an Agrobacterium tumefaciens-mediated method. An improved genetic transformation rate (12‰) was obtained when the explants were pre-cultured for 3 days, immersed in bacterial suspension (OD600≈0.8) for 5 min, and co-cultivated for 5 days. The binary vector pBI121 containing Zm401, a maize pollen-specific gene, was introduced into the Agrobacterium strain LBA4404 and transformed into the explants using the genetic transformation system. Gene integration into the lily genome was confirmed by polymerase chain reaction (PCR) and PCR–Southern analysis. These results could lead to the production of new pollenless lily plants.

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