葡萄根癌生防菌株E26生防效果的离体检测方法研究.pdfVIP

中国农业科学 2007,40(7):1395-1402 Scientia Agricultura Sinica 葡萄根癌生防菌株E26生防效果的离体检测方法研究 王 靖，郭岩彬，王建辉，李金云，王慧敏 （中国农业大学植物病理学系，北京 100094 ） 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 1/2 B5 培养基的基础上改良出适合于葡萄离体茎段培养用的 MSC 1/2B5 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。 关键词：葡萄根癌病；生防效果；离体检测 A Comparative Study of Testing Methods for the Effect of Biocontrolling Agrobacterium vitis E26 Against Grape Crown Gall in vitro WANG Jing, GUO Yan-bin, WANG Jian-hui, LI Jin-yun, WANG Hui-min （Department of Plant Pathology, China Agricultural University, Beijing 100094 ） Abstract: 【Objective 】To 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. 【Method 】By 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