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【目的】从农杆菌介导获得的灰葡萄孢Rose BC-3的突变体库中筛选侵染垫缺失突变体菌株,并明确其相关生物学特性。【方法】将菌株接种于洋葱表皮,利用棉兰染色观察侵染垫形成情况,筛选得到一个侵染垫缺失突变体(AT19)。采用形态学方法、离体叶片接种法、钌红染色法、小麦种子幼芽生长抑制法分别对该菌株的菌落培养性状、侵染垫产生情况、致病力、产果胶酶能力以及产植物毒性代谢产物能力进行测定。【结果】筛选灰葡萄孢突变体168株,根据侵染垫形成可分为三类:快速形成侵染垫型(158株)、缓慢形成侵染垫型(9株)和侵染垫形成缺陷型(1株,AT19)。AT19在接种洋葱120 h后依然无法形成成熟侵染垫。该菌株生长较为缓慢,菌落扩展均匀,可以产生分生孢子,对烟草、草莓、蚕豆和豌豆叶片均不能致病,可以产生果胶酶和植物代谢毒性物质。【结论】突变体菌株AT19可以产生果胶酶和植物代谢毒性物质,其致病力缺失与侵染垫产生缺陷相关。研究结果为了解灰葡萄孢侵染垫形成分子机制提供基础材料。
【Objective】 The objective of this study was to screen out mutant strains of mutant of Rose BC-3 from Agrobacterium tumefaciens and identify the related biological characteristics. 【Method】 The strain was inoculated on the onion epidermis, the formation of infection pad was observed by Medan staining, and a mutant with an infection pad deletion (AT19) was screened out. Morphological methods, in vitro leaf inoculation method, ruthenium red staining method, wheat seedling growth inhibition method of the strain colony culture traits, infection pad production, pathogenicity, pectinase production capacity and plant Toxic metabolites were measured for their ability. 【Result】 A total of 168 strains of Botrytis cinerea mutants were screened. According to the formation of infection mats, there are three types: rapid formation of infection mats (158 strains), slow formation of infection mats (9 strains) and infection mats formation defects Type (1 strain, AT19). AT19 still can not form mature infiltration mat after inoculation of onion for 120 h. The strain grows more slowly, the colonies spread evenly, can produce conidia, tobacco, strawberry, broad bean and pea leaves are not pathogenic, can produce pectinase and plant metabolic toxic substances. 【Conclusion】 Mutant strain AT19 can produce pectinase and plant metabolic toxic substances, and its lack of virulence is related to the defect of infection pad. The results provide the basic material for understanding the molecular mechanism of the formation of Botrytis cinerea infection mats.