香蕉枯萎镰刀菌致病性快速测定方法的建立

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建立致病性变异突变体库是研究香蕉枯萎镰刀菌致病机理的有效方法,此方法需要对大量突变体进行致病性测定,经典的根部接种测定方法耗时长达40 d,工作量十分巨大。因此,建立一种简便快速的致病性测定方法是高效建立香蕉枯萎病菌致病性变异突变体库的关键。本研究以香蕉巴西蕉和粉蕉叶片为材料,采用香蕉离体叶片分别接种香蕉枯萎病菌1号小种、4号小种、致病性丧失突变体、致病性严重减弱突变体,分别置于20℃、25℃、30℃、35℃、37℃条件下进行保湿培养3 d、5 d、7 d,观察发病情况,测定病斑大小。并通过根部接种法对离体叶片接种法的结果进行验证。结果表明,离体叶片测定法和根部接种法测定的结果一致,最佳致病温度30℃。从而建立了一种简便快速的香蕉枯萎病突变体致病性测定方法(离体叶片接种法)。运用该法在适宜的条件下可以准确、可靠、快速和简便测出香蕉枯萎病菌致病性,大大提高了突变体致病性测定筛选的效率。 The establishment of a pathogenic mutant library is an effective method to study the pathogenic mechanism of Fusarium solani in banana. This method requires pathogenicity determination of a large number of mutants. The classical method of root inoculation assay takes up to 40 days and the workload is huge . Therefore, establishing a simple and rapid pathogenicity determination method is the key to establish efficient mutant library of Fusarium wilt pathogen. In this study, Banana and Banana banana leaves were used as materials. The banana leaves were inoculated with Fusarium oxysporum f. Sp. # 1, # 4, pathogenicity mutants and pathogenic severely weakened mutants, respectively Moisture culture was carried out at 20 ℃, 25 ℃, 30 ℃, 35 ℃, 37 ℃ for 3 days, 5 days and 7 days respectively. The incidence of lesions was measured and the lesion size was determined. The results of inoculation method of leaves in vitro were verified by root inoculation method. The results showed that the results of in vitro leaf assay and root inoculation method were consistent, and the optimal pathogenicity temperature was 30 ℃. Thus, a simple and rapid method for determination of pathogenicity of banana wilt disease mutant (in vitro leaf inoculation method) was established. The method can be used to detect the pathogenicity of Fusarium oxysporum fumigatus accurately, reliably, rapidly and simply under the appropriate conditions and greatly improve the efficiency of the mutant pathogenicity test.
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