灰葡萄孢致病基因BcKMO与病菌MAPK信号途径的关系  

Analysis of the Relationship Between BcKMO Gene and MAPK Signaling Pathway in Botrytis cinerea

作　　者：苏有科 袁雪梅 王敏[1] 藏金萍[1] 曹宏哲 张康 董金皋[1] 张靖[1] 邢继红[1] SU Youke;YUAN Xuemei;WANG Min;ZANG Jinping;CAO Hongzhe;ZHANG Kang;DONG Jingao;ZHANG Jing;XING Jihong(Key Laboratory of Hebei Province for Plant Physiology and Molecular Pathology,Mycotoxin and Molecular Plant Pathology Laboratory of Hebei Agricultural University,Baoding 071001,China)

机构地区：[1]河北省植物生理与分子病理学重点实验室,河北农业大学真菌毒素与植物分子病理学实验室,河北保定071001

出　　处：《华北农学报》2018年第5期89-94,共6页Acta Agriculturae Boreali-Sinica

基　　金：河北省自然科学基金项目(C2018204045);河北省高等学校科学技术研究项目(ZD2016001);河北省留学人员科技活动择优资助项目(0316012);2018年大学生创新创业训练计划项目(201810086064)

摘　　要：为明确灰葡萄孢致病基因BcKMO与病菌MAPK信号途径之间的关系,利用MAPK信号途径特异性的抑制剂U0126处理灰葡萄孢野生型菌株BC22、BcKMO基因的T-DNA插入突变体BCG183和回复突变体BCG183/BcKMO。结果发现,突变体BCG183对抑制剂U0126不敏感,受抑制的程度显著低于野生型和回复突变体。对BcKMO基因与MAPK信号途径关键基因bmp1和bmp3的表达规律进行分析,结果发现,BcKMO与bmp3均是在7 d的菌丝和菌核中表达水平较高,BcKMO与bmp1、bmp3基因均是在含蔗糖和果糖培养基上培养的病菌中表达水平较高。利用Real-time PCR技术,检测BcKMO基因突变对bmp1、bmp3基因表达的影响以及bmp1、bmp3基因突变对BcKMO基因表达的影响,结果发现,突变体BCG183中bmp1的表达水平均显著高于野生型和回复突变体,bmp3的表达水平显著低于野生型和回复突变体; bmp1的RNAi突变体中BcKMO基因表达水平显著高于野生型,而bmp3基因的RNAi突变体中BcKMO基因表达水平显著低于野生型。结果表明,BcKMO基因负调控bmp1的表达,正调控bmp3的表达;而bmp1负调控BcKMO基因的表达,bmp3正调控BcKMO基因的表达。研究结果可为阐明灰葡萄孢BcKMO基因调控病菌生长、发育和致病力的机制奠定基础。To analyzed the relationship between BcKMO gene and MAPK signaling pathway in Botrytis cinerea,the specific inhibitor of MAPK signaling pathways U0126 was used to detect the sensitivity of the BcKMO gene mutants.It was found that the sensitivity of the BCG183 mutant to U0126 was remarkably weak compared to that of the BC22 and BCG183/BcKMO.Expression patterns of BcKMO and MAPK signaling pathway key genes bmp1 and bmp3 were detected.The expression levels of BcKMO and bmp3 were stronger in 7 d mycelia and sclerotia of BC22.The expression levels of BcKMO,bmp1,and bmp3 were stronger in BC22 cultured on medium with sucrose or fructose.Real-time PCR technology was used to detect the expression levels of bmp1 and bmp3 in the BcKMO gene mutants,and the BcKMO expression levels in RNAi mutant of bmp1 and bmp3.The expression level of bmp1 was significantly upregulated in the BCG183 mutant,the expression level of bmp3 was significantly downregulated in the BCG183 mutant.The expression level of BcKMO in the RNAi mutants of bmp1 gene was obviously higher than that of BC22;the expression level BcKMO gene in the RNAi mutants of bmp3 was obviously lower than that of BC22.These results indicated that BcKMO negatively regulated the expression of bmp1 and positively regulated the bmp3 expression,bmp1 negatively regulated the BcKMO gene expression,and bmp3 positively regulated the BcKMO gene expression.Therefore,the results provide a foundation for clarifying the molecular mechanism of the BcKMO gene in growth,development and pathogenicity in Botrytis cinerea.

关 键 词：灰葡萄孢 BcKMO MAPK信号途径 

分 类 号：Q78[生物学—分子生物学] S432.1[农业科学—植物病理学]