Functional characterization of CgPBS(2), a MAP kinase kinase in Colletotrichum gloeosporioides, using osmotic stress sensitivity as a selection marker

文献类型： 外文期刊

第一作者： Lin, Chunhua

作者： Lin, Chunhua;Zheng, Fucong;Miao, Weiguo;Lin, Chunhua;Huang, Guixiu

作者机构：

关键词： CgPBS(2); MAP kinase kinase; Osmotic stress; Selection marker; Colletotrichum gloeosporioides; Hevea brasiliensis

期刊名称：EUROPEAN JOURNAL OF PLANT PATHOLOGY （ 影响因子：1.907； 五年影响因子：2.022 ）

ISSN： 0929-1873

年卷期： 2018 年 152 卷 3 期

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收录情况： SCI

摘要： Colletotrichum leaf disease of Hever brasiliensis (rubber tree) caused by C. gloeosporioides is one of the major causes of declining rubber tree yields. Little is known about the fungal molecular characters that are important for pathogenicity on rubber tree and fungicide resistance. In this study, we cloned the CgPBS(2) gene, the key component of the Hog1 pathway which controls various aspects of osmoregulation and fungicide resistance in various fungal pathogens, including the causal agent of Colletotrichum leaf disease of rubber tree. We characterized the function of the CgPBS(2) gene by reverse genetics. Because the Hog1 pathway plays an important role in stress responses, we obtained a CgPBS(2) gene deletion mutant by PEG-mediated transformation of protoplasts after reducing the concentration of sucrose in the screening medium from 1.0M to 0.2M. Then, the complemented transformants and GFP-labelled CgPBS(2) gene transformants were selected directly under highly hyperosmotic medium (PDA+1.5M sorbitol) without using other selectable gene markers. Phenotypic observations showed that the CgPBS(2) protein was mainly localized in the conidial cytoplasm of the CgPBS(2)-GFP transformants. In addition, disruption of CgPBS(2) led to sensitivity to hyperosmosis and high salt concentration as well as resistance to the fungicide fludioxonil. No obvious difference in virulence was observed between the null mutant and the wild-type strain. These results provide insights into the role of the CgPBS(2) gene in osmotic stress, salt stress and fludioxonil resistance and suggest that osmotic stress sensitivity can be used as a selection marker.

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