scholarly journals Adenine Auxotrophic Mutants ofAspergillus oryzae: Development of a Novel Transformation System with Triple Auxotrophic Hosts

2004 ◽  
Vol 68 (3) ◽  
pp. 656-662 ◽  
Author(s):  
Feng Jie JIN ◽  
Jun-ichi MARUYAMA ◽  
Praveen Rao JUVVADI ◽  
Manabu ARIOKA ◽  
Katsuhiko KITAMOTO
Keyword(s):  
Transformation System ◽  
Auxotrophic Mutants

Development of an Agrobacterium-mediated Transformation System for `Beurre Bosc' Pear

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 461d-461
Author(s):  
Richard L. Bell ◽  
Ralph Scorza ◽  
Chinnathambi Srinivasan
Keyword(s):  
Shoot Proliferation ◽  
Induction Medium ◽  
Shoot Induction ◽  
Transformation System ◽  
Gus Histochemical Assay ◽  
Young Leaves ◽  
Leaf Tissues ◽  
Shoot Induction Medium ◽  
Basal Salts

An efficient regeneration/transformation system was developed for `Beurre Bosc' pear. Young leaves were harvested from in vitro shoots proliferated on a medium containing MS basal salts and 5 BAP, 0.5 μM IBA, and 0.6M3. Shoot regeneration was optimized using a modification of the medium of Chevreau and Leblay (1993). Explants were cultured on shoot induction medium contained 10 μM TDZ and 1 μM IBA for 4 weeks in the dark, and then transfered to a similar, but auxinless, regeneration medium until shoots developed, usually after an additional 4 to 8 weeks. Leaf tissues were transformed by co-cultivation for 3 days with Agrobacterium tumefaciens EHA101 carrying a pGA482 plasmid containing NPTII, GUS, and rolC genes, followed by cultivation on SIM containing 300 mg/L timentin. Putative transgenic plants were selected on shoot induction medium containing 80mg/L kanamycin, and multiplied on shoot proliferation medium. Four clones were confirmed as transgenic using the GUS histochemical assay and Southern blots for the NPTII and rolC genes. Plants of each clone have been rooted and successfully transfered to the greenhouse for further analysis of gene expression.

An efficient transient transformation system for gene function studies in pumpkin (Cucurbita moschata D.)

2021 ◽  
Vol 282 ◽  
pp. 110028
Author(s):  
Xuejin Chen ◽  
Songtao He ◽  
Lina Jiang ◽  
Xinzheng Li ◽  
Weili Guo ◽  
...  
Keyword(s):  
Gene Function ◽  
Transient Transformation ◽  
Cucurbita Moschata ◽  
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.

scholarly journals Biochemical Aspects of Competence in the Bacillus subtilis Transformation System

1963 ◽  
Vol 238 (9) ◽  
pp. 3119-3125 ◽  
Author(s):  
F.E. Young ◽  
John Spizizen ◽  
I.P. Crawford
Keyword(s):  
Bacillus Subtilis ◽  
Transformation System

scholarly journals Survival and mutant production induced by mutagenic agents in Metarhizium anisopliae

1995 ◽  
Vol 52 (3) ◽  
pp. 548-554 ◽  
Author(s):  
V. Kava - Cordeiro ◽  
E.A. Luna - Alves - Lima ◽  
J.L. Azevedo
Keyword(s):  
Gamma Radiation ◽  
Ultraviolet Light ◽  
Metarhizium Anisopliae ◽  
Entomopathogenic Fungus ◽  
Nitrous Acid ◽  
Wild Strain ◽  
Wild Type ◽  
Survival Curves ◽  
Auxotrophic Mutants ◽  
Morphological Mutants

A wild strain of Metarhizium anisopliae, an entomopathogenic fungus, was submitted to three mutagenic agents: gamma radiation, ultraviolet light and nitrous acid. Survival curves were obtained and mutants were selected using different mutagenic doses which gave 1 to 5% survival. Morphological and auxotrophic mutants were isolated. Morphological mutants were grouped in a class with yellow conidia and other with pale vinaceous conidia as opposed to the green wild type conidia. Auxotrophic mutants had requirements for vitamin and aminoacid biosynthesis. More than 58% of the total auxotrophk mutants required proline/aipnine. Gamma radiation showed to be the most efficient mutagenic agent giving 0.2% of auxotrophk mutants followed by ultraviolet light (0.12%) and nitrous acid (0.06%).The conidial colour and auxotrophk mutants isolated until now from M. anisopliae were reviewed.

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