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The Ifchit1 chitinase gene acts as a critical virulence factor in the insect pathogenic fungus Isaria fumosorosea

  • Applied genetics and molecular biotechnology
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Abstract

The filamentous fungus, Isaria fumosorosea, is a promising insect biological control agent. Chitinases have been implicated in targeting insect cuticle structures, with biotechnological potential in insect and fungal control. The I. fumosorosea chitinase gene, Ifchit1, was isolated and determined to encode a polypeptide of 423 amino acids (46 kDa, pI = 6.53), present as a single copy in the I. fumosorosea genome. A split marker transformation system was developed and used to construct an Ifchit1 gene knockout. The ΔIfchit1 strain displayed minor alterations in mycelial growth on diverse media at 26 °C compared to the wild type and complemented (ΔIfchit1::Ifchit1) strains; however, colony morphology was affected, and the mutant strain had a temperature sensitive phenotype (32 °C). Although sporulation was delayed for the mutant, overall conidial production was almost twice than that of wild type. Biochemical assays indicated decreased chitinase activity during growth in Czapek-Dox liquid media for the ΔIfchit1 strain. Insect bioassays using diamondback moth, Plutella xylostella, larvae revealed decreased infectivity, i.e., increased LC 50 (threefold to fourfold) and a significantly delayed time to death, LT 50 from 3 to 6 days, for the ΔIfchit1 strain compared to the wild type and complemented strains. These data indicate an important role for the Ifchit1 chitinase as a virulence factor in I. fumosorosea.

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Funding

This research was funded by grants from the “863” Program of China (2011AA10A204-5), the National Natural Science Foundation of China (31170391), and the National Science Foundation grant IOS-1121392 to NOK.

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Authors and Affiliations

  1. Engineering Research Center of Biological Control, Ministry of Education, College of Agriculture, South China Agricultural University, Guangzhou, 510642, China

    Zhen Huang, Yongfen Hao, Tianni Gao, Yü Huang & Shunxiang Ren

  2. Department of Microbiology and Cell Science, Institute of Food and Agricultural Sciences, University of Florida, Bldg 981, Museum Rd, Gainesville, FL, 32611, USA

    Zhen Huang & Nemat O. Keyhani

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  1. Zhen Huang
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  2. Yongfen Hao
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  3. Tianni Gao
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  4. Yü Huang
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  6. Nemat O. Keyhani
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Correspondence to Zhen Huang, Shunxiang Ren or Nemat O. Keyhani.

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Yongfen Hao, Joint first author.

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Huang, Z., Hao, Y., Gao, T. et al. The Ifchit1 chitinase gene acts as a critical virulence factor in the insect pathogenic fungus Isaria fumosorosea . Appl Microbiol Biotechnol 100, 5491–5503 (2016). https://doi.org/10.1007/s00253-016-7308-z

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