【摘要】 【目的】筛选葡萄离体茎段和叶片的最适表面消毒条件、离体培养基和接种方法,建立生防菌株E26防治葡萄根癌病效果的离体检测方法。【方法】通过比较6种表面消毒方法,4种培养基和2种接种方法,建立最优的E26生防效果离体检测方法;并通过与温室、大田生防效果的比较以及瘤组织切片观察、冠瘿碱纸电泳检测验证该离体检测方法的准确性。【结果】在1/2B5培养基的基础上改良出适合于葡萄离体茎段培养用的MSC1/2B5培养基;筛选出葡萄离体茎段的最佳消毒条件和接种方法,建立了检测E26生防效果的离体茎段接种法。该方法的检测结果与葡萄活体植株的检测结果基本一致,能够快速准确地反映E26的防效;并且,瘤组织切片观察和冠瘿碱纸电泳检测结果证实了该离体检测方法的可靠性。叶圆片法检测时,根癌瘤只能在葡萄叶圆片的叶脉处形成,瘤组织较小,难以准确反映E26的生防效果。【结论】本研究建立的离体茎段检测E26的生防效果是一种比较理想的离体检测E26生防效果的方法。更多还原

【Abstract】 ObjectiveTo determine suitable surface sterilization methods,media and inoculation methods for grape stem and leaf explants and then establish testing methods for the effect of biocontrol Agrobacterium vitis E26 against grape crown gall in vitro. MethodBy means of comparing effects of six sterilization methods on stem and leaf explants and effects of four different media on explants’ development,and evaluating individual biocontrol effect of two methods for E26 inoculating on stem and leaf explants,we attempt to acquire testing methods for biocontrol effect of E26. Meanwhile,by the comparison of biocontrol effects in vitro with that in greenhouse and field tests,observation of tumor tissue morphology,and detection of opine in tumors through paper electrophoresis,the veracity of this acquired testing method for biocontrolling effect of E26 was to be confirmed.ResultMSC 1/2B5 medium,suited for culture of grape stem in vitro ,was achieved with modification of 1/2B5 medium. The best methods of sterilization and inoculation were also screened in this experiment. The stem explants in vitro test method was established and it was demonstrated the same effect when compared with the in vivo tests,and could be the quick and accurate evaluation to indicate the effect of E26. Observation of tissue morphology and detection of opine in tumors confirmed the veracity of this esting method While the grape leaf in vitro test could not accurately reflect the effect of E26,because crown galls formed only on leaf veins and were too small to be examined. ConclusionThe grape stem in vitro test can be a promising method to detect the effect of E26.更多还原