水稻离体叶片抗纹枯病接种方法的研究  

Studies on the detached-leaf inoculation method for determination of rice resistance to sheath blight(Rhizoctonia solani)

作　　者：马晨燕[1,2] 袁正杰[2] 杨海河[1,2] 曲海艳 何海燕[2] 张玉[2] 瞿绍洪[2] 

机构地区：[1]浙江师范大学化学与生命科学学院,浙江金华321004 [2]浙江省农业科学院病毒学与生物技术研究所,浙江杭州310021

出　　处：《浙江农业学报》2016年第10期1730-1737,共8页Acta Agriculturae Zhejiangensis

基　　金：转基因生物新品种培育重大专项(2012ZX08009001);浙江省农业科学院科技创新能力提升工程项目(2015CX07);浙江省农业科学院省部共建国家重点实验室培育基地开放基金(2010DS700124-KF1210;2010DS700124-KF1406)

摘　　要：水稻纹枯病是影响水稻生产的重要病害之一。水稻纹枯病抗性属于数量遗传性状,目前尚未发现高抗或免疫的水稻种质材料。为了提高水稻抗纹枯病种质的筛选和研究效率,对水稻离体叶片抗纹枯病接种的实验条件进行了更精确的控制,并采用了量化的病情调查方法。取纹枯菌接种离体水稻叶片组织进行qRT-PCR分析,水稻病程相关基因呈诱导表达,验证了纹枯病菌对水稻的侵染。采用离体叶片接种、大田成株期接种、苗期"微室"接种法,对抗纹枯病水稻品种和敏感品种进行测试。抗性品种和敏感品种纹枯病病级的统计检验呈显著差异,3种方法的结果表现一致。改进的离体叶片接种方法具有易于操作和重复性好的优点,适用于大规模筛选抗纹枯病水稻种质材料。Rice sheath blight（ Rhizoctonia solani） is one of the important diseases causing large loss to rice production. Rice resistance to R. solani belongs to quantitative genetic trait. Rice germplasm with high resistance or immunity to sheath blight has not yet been identified. In order to improve the efficiency of screening and characterizing sheath blight-resistant rice germplasm,we made precise control of the experimental conditions for the detached-leaf inoculation method,and developed a quantitative protocol for the evaluation of disease severity. The detached leaf tissues inoculated with R. solani were subjected to quantitative reverse-transcription qRT-PCR analysis. The induced expression of rice pathogenesis-related genes verified the infection of the pathogen into rice. A sheath blight-resistant rice cultivar and a sensitive cultivar were tested using the detached-leaf method,the field inoculation method and the micro-chamber inoculation method,respectively. The three methods showed similar results,and disease scores of the resistant and sensitive cultivars were significantly different based on statistical tests. Taken together,the improveddetached-leaf inoculation method had the advantages of easy operation and good repeatability,and could be used for large-scale screening of sheath blight-resistant rice germplasm.

关 键 词：水稻 纹枯病 RHIZOCTONIA SOLANI 离体叶片法 病程相关基因 

分 类 号：S511[农业科学—作物学] S41-30