利用瞬间表达技术分析小麦几丁质酶和β-1,3葡聚糖苷酶基因的抗病性功能  

Disease-resistance Function Analysis of Wheat Chitinase and β-1,3-Glucanase Genes by a Transient Expression System

作　　者：王华忠[1] 邢莉萍[1] 李万隆[1] 陈佩度[1] 

机构地区：[1]南京农业大学作物遗传与种质创新国家重点实验室,江苏南京210095

出　　处：《麦类作物学报》2006年第1期7-12,共6页Journal of Triticeae Crops

基　　金：国家高技术研究发展计划(863)项目(2001AA222152;2004AA222140)

摘　　要：几丁质酶基因和-β1,3葡聚糖苷酶基因是植物抗病反应过程中的两类重要的病程相关蛋白(PR)基因。为了快速准确地评价此类基因组成性表达后的抗病效果及在分子育种上的应用价值,利用瞬间表达技术分别在感病小麦品种离体叶片表皮细胞中过量表达1个小麦几丁质酶基因Cht4和2个小麦-β1,3葡聚糖苷酶基因G lb3s2和G lb3s6,并以GUS基因的共表达标识阳性转化细胞。基因枪轰击后高密度接种白粉病菌孢子,40h后观察转化阳性表皮细胞及其表面孢子的发育,并以阳性转化细胞中白粉病菌成功侵入的细胞所占比例(侵入频率)为指标分析目标基因的表达对白粉病菌入侵及吸器形成产生的影响(对照为单独导入GUS基因的表皮细胞)。结果表明,小麦几丁质酶基因Cht4(侵入频率为18.3%,对照为25.04%)和小麦-β1,3葡聚糖苷酶基因G lb3s2(侵入频率为19.63%,对照为24.63%)在感病小麦品种叶片表皮细胞中的瞬间表达,对白粉病菌侵入和吸器形成均有抑制作用,在一定程度上增强了表达细胞对白粉病菌的抗性。Chitinase and β-1,3-Glucanase Genes are two important classes of PR genes functioning in plant-pathogen interaction. In order to evaluate their disease-resistance function after constitutive expression rapidly, one wheat ehitinase gene Cht4 and two wheatβ-1,3-glucanase genes Glb3s2 and Glb3s6 were respectively over expressed in powdery mildew-susceptible wheat variety detached leaf epidermal cells by a transient expression system. GUS gene was co-transformed with the target gene to mark the transformed cells. After transformation, leaf surface was heavily inoculated with powdery mildew fungus Blumeria graminis f. sp. tritici conidiospores. 40 h after inoculation, penetration of the fungus and formation of haustoria in transformed cells were observed to evaluate the effect of the target gene＇s product on the invasion of the fungus. The results implied that two of these three genes, Glb3s2 and Cht4, when transiently expressed in leaf epidermal cells of susceptible wheat variety, can partially inhibit penetration of conidiospores and formation of haustoria, and to some extent increase the resistance of expression cells to powdery mildew.

关 键 词：小麦 白粉病 瞬间表达 几丁质酶基因 β-1 3葡聚糖苷酶基因 

分 类 号：S512.1[农业科学—作物学] S336