URA3 as a Selectable Marker for Disruption and Functional Assessment of PacC Gene in the Entomopathogenic Fungus Isaria javanica

文献类型： 外文期刊

第一作者： Zou, Manling

作者： Zou, Manling;Xin, Bei;Sun, Xin;Lin, Runmao;Lu, Junru;Qi, Jing;Cheng, Xinyue;Xin, Bei;Lin, Runmao;Lu, Junru;Xie, Bingyan;Cheng, Xinyue

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关键词： entomopathogenic fungus; Isaria javanica; ura3 gene; uridine auxotrophy; markerless transformation; PacC gene; the pH response transcription factor

期刊名称：JOURNAL OF FUNGI （ 影响因子：4.7； 五年影响因子：5.2 ）

ISSN：

年卷期： 2023 年 9 卷 1 期

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收录情况： SCI

摘要： An effective selection marker is necessary for genetic engineering and functional genomics research in the post-genomic era. Isaria javanica is an important entomopathogenic fungus with a broad host range and prospective biocontrol potentials. Given that no antibiotic marker is available currently in this fungus, developing an effective selection marker is necessary. In this study, by applying overlap PCR and split-marker deletion strategy, combining PEG-mediated protoplasm transformation method, the uridine auxotrophy gene (ura3) in the I. javanica genome was knocked out. Then, using this transformation system, the pH response transcription factor gene (IjpacC) was disrupted successfully. Loss of IjpacC gene results in an obvious decrease in conidial production, but little impact on mycelial growth. The virulence of the Delta IjpacC mutant on caterpillars is similar to that of the wild-type strain. RT-qPCR detection shows that expression level of an acidic-expressed S53 gene (IF1G_06234) in Delta IjpacC mutant is more significantly upregulated than in the wild-type strain during the fungal infection on caterpillars. Our results indicate that a markerless transformation system based upon complementation of uridine auxotrophy is successfully developed in I. javanica, which is useful for exploring gene function and for genetic engineering to enhance biological control potential of the fungus.

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