Relationship between expression level of hygromycin B-resistant gene and<i>Agrobacterium tumefaciens</i>-mediated transformation efficiency in<i>Beauveria bassiana</i>JEF-007

Nai, Yu‐Shin; Lee, M.R.; Kim, Seokhyeon; Lee, S.J.; Kim, J.C.; Yang, Yiting; Kim, J.S.

doi:10.1111/jam.13529

Cited by 7 publications

(2 citation statements)

References 20 publications

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“…In present study, the C. lindemuthianum transformants showed different resistance levels to hygromycin B which could possibly be attributed to integration site of T‐DNA leading to chromosomal positioning effect or integration of different T‐DNA copies (Nai et al . 2017). Eighty‐four per cent of the transformants showed single copy integrations suggesting that chromosomal positioning has predominant effect on differential response to hygromycin B.…”

Section: Resultsmentioning

confidence: 99%

Optimizing the Agrobacterium tumefaciens-mediated transformation conditions in Colletotrichum lindemuthianum: a step forward to unravel the functions of pathogenicity arsenals

Nabi

Banoo

Rasool

et al. 2022

Letters in Applied Microbiology

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Significance and Impact of the Study: In the present study, we for the first time, report an efficient Agrobacterium tumefaciens-mediated transformation (ATMT) system for a hemibiotrophic fungus, Colletotrichum lindemuthianum. This optimized ATMT protocol is versatile and can be used for the functional genetic studies. Insertional mutagenesis studies produce thousands of mutant lines that need to be evaluated for pathogenicity loss and the traditional methods of pathogenicity assay in bean-anthracnose pathosystem are time consuming and laborious. Hence, we developed a quick and robust screening method that produce consistent results similar to the established bioassay methods. Our optimized ATMT and virulence analysis protocols will prove useful in understanding the bean-anthracnose interface.

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Section: Resultsmentioning

confidence: 99%

Optimizing the Agrobacterium tumefaciens-mediated transformation conditions in Colletotrichum lindemuthianum: a step forward to unravel the functions of pathogenicity arsenals

Nabi

Banoo

Rasool

et al. 2022

Letters in Applied Microbiology

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“…The first step in establishing the transformation system is to determine the appropriate dominant selection marker. Among selection markers, hygromycin B, Geneticin (G418), and phleomycin are widely used for the establishment of fungal transformation systems (Campbell et al, 2002;Shao et al, 2015;Nai et al, 2017;He et al, 2020). At present, the genetic transformation systems of fungi in the genus Cordyceps mainly include the Agrobacterium-mediated genetic transformation system and the polyethylene glycol (PEG)-mediated protoplast genetic transformation system (Huang et al, 2016;Lou et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

Development of Cordyceps javanica BE01 with enhanced virulence against Hyphantria cunea using polyethylene glycol-mediated protoplast transformation

Wang

Chen

2022

Front. Microbiol.

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Cordyceps javanica has promising application prospects as an entomopathogenic fungus with a wide range of hosts. To enhance the virulence of C. javanica, a polyethylene glycol (PEG)-mediated protoplast genetic transformation system was constructed. Strains overexpressing the subtilisin-like protease genes CJPRB and CJPRB1 and the tripeptidyl peptidase gene CJCLN2-1 were constructed with this system, and the effects of these strains on Hyphantria cunea were tested. The aminoglycoside G418 was used at 800 μg ml−1 to screen the transformants. C. javanica hyphae were degraded with an enzyme mixture to obtain protoplasts at 1.31 × 107 protoplasts ml−1. The transformation of 2 μg of DNA into 1,000 protoplasts was achieved with 20% PEG2000, and after 6 h of recovery, the transformation efficiency was 12.33 ± 1.42 transformants μg−1 plasmid. The LT50 values of CJPRB, CJPRB1, and CJCLN2-1-overexpressing C. javanica strains were 1.32-fold, 2.21-fold, and 2.14-fold higher than that of the wild-type (WT) strain, respectively. The three overexpression strains showed no significant differences from the WT strain in terms of colony growth, conidial yield, and conidial germination rate. However, the infection rate of the CJPRB1 strain was faster than that of the WT strain, with infection occurring within 4–5 days. The CJCLN2-1 strain had a significantly higher mortality rate than the WT strain within 4–10 days after infection. A C. javanica genetic transformation system was successfully constructed for the first time, and an overexpression strain exhibited enhanced virulence to H. cunea compared with the WT strain.

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Improved production of β-glucan by a T-DNA–based mutant of Aureobasidium pullulans

Chen

Wang

et al. 2021

Appl Microbiol Biotechnol

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Relationship between expression level of hygromycin B-resistant gene andAgrobacterium tumefaciens-mediated transformation efficiency inBeauveria bassianaJEF-007

Abstract: These results provide useful information for determining AtMT effectiveness in B. bassiana isolates, particularly antibiotic susceptibility and the role of promoters.

Cited by 7 publications

References 20 publications

Optimizing the Agrobacterium tumefaciens-mediated transformation conditions in Colletotrichum lindemuthianum: a step forward to unravel the functions of pathogenicity arsenals

Optimizing the Agrobacterium tumefaciens-mediated transformation conditions in Colletotrichum lindemuthianum: a step forward to unravel the functions of pathogenicity arsenals

Development of Cordyceps javanica BE01 with enhanced virulence against Hyphantria cunea using polyethylene glycol-mediated protoplast transformation

Improved production of β-glucan by a T-DNA–based mutant of Aureobasidium pullulans

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