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Establishment of polyethylene‐glycol‐mediated protoplast transformation for Lecanicillium lecanii and development of virulence‐enhanced strains against Aphis gossypii
Author(s) -
Zhang YanJun,
Xie Ming,
Zhang XiaoLin,
Peng DeLiang,
Yu WenBin,
Li Qiang,
Li Qian,
Zhao JinJin,
Zhang ZhaoRong
Publication year - 2016
Publication title -
pest management science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.296
H-Index - 125
eISSN - 1526-4998
pISSN - 1526-498X
DOI - 10.1002/ps.4236
Subject(s) - protoplast , biology , aphis gossypii , biopesticide , mycelium , beauveria bassiana , transformation (genetics) , botany , microbiology and biotechnology , horticulture , biological pest control , pest analysis , pesticide , gene , agronomy , aphididae , biochemistry , homoptera
BACKGROUND Lecanicillium lecanii has been developed as a biopesticide and used in biological control of several agricultural insects. To improve fungal virulence, an optimised polyethylene glycol ( PEG )‐mediated protoplast transformation system was established for L. lecanii . Pr1A ‐like cuticle‐degrading protease gene ( Cdep1 ) from Beauveria bassiana was transferred into L. lecanii , and its resulting activity against Aphis gossypii was assessed. RESULTS The optimised protoplast generation yielded 2.5 × 10 8 protoplasts g −1 wet mycelium of fungi, and gave nearly 98% viability and 80% regeneration on plates. Protease activities were increased about fivefold in transformants expressing CDEP1 . The median lethal concentration ( LC 50 ) for transformants expressing CDEP1 was twofold lower than that for the wild type ( WT ). The median survival time ( LT 50 ) for transformants expressing CDEP1 was also 14.2% shorter than that for WT , though no significant difference. There were no significant differences in conidial germination as colony growth and conidial yield on plates between transformants expressing CDEP1 and WT . The transformants expressing CDEP1 grew significantly quicker than WT in insects. The transformants expressing CDEP1 were lower in conidial yields on insect cadavers, but insignificantly different from WT . CONCLUSION The PEG ‐mediated protoplast transformation system was effective for L . lecanii , and the expression of CDEP1 significantly enhanced fungal virulence against cotton aphids. © 2016 Society of Chemical Industry