甘蔗几丁质酶基因ScChiⅦ1的克隆与表达分析  

Molecular Cloning and Expression Analysis of Chitinase Gene ScChiⅦ1 in Sugarcane

作　　者：王珊珊[1] 苏亚春[1] 杨玉婷[1] 郭晋隆[1] 许莉萍[1] 

机构地区：[1]福建农林大学农业部福建甘蔗生物学与遗传育种重点实验室,福建福州350002

出　　处：《热带作物学报》2014年第2期289-298,共10页Chinese Journal of Tropical Crops

基　　金：现代农业产业技术体系建设专项资金(No.CARS-20)

摘　　要：以甘蔗(Saccharum spp.hybrid)抗黑穗病基因型崖城05-179为材料,采用同源克隆法获得了1个几丁质酶基因全长cDNA序列(GenBank登录号为KF279661),命名为ScChiⅦ1,序列长907 bp,编码299个氨基酸,属于糖苷水解酶第19家族,预测为胞外分泌蛋白且兼具溶菌酶活性。聚类分析显示,该基因与其他植物ClassⅦ几丁质酶基因聚为一类。基因组序列(GenBank登录号为KF279662)分析显示,该基因含有2个长度分别为615 bp和86 bp的内含子。实时荧光定量分析结果表明,甘蔗黑穗病菌胁迫下,ScChiⅦ1在抗病(崖城05-179)和感病基因型(柳城03-182)中差异表达,推测ScChiⅦ1可能参与黑穗病菌胁迫的防御反应。组织特异性表达分析结果显示,ScChiⅦ1在甘蔗叶、芽、皮、肉、根中均有表达,但在芽中表达量最高,推测与该病原从蔗芽侵入有关。研究结果为进一步研究甘蔗几丁质酶基因功能及甘蔗与黑穗病菌互作机制提供了有益的信息。Using homologous cloning method, one full length chitinase cDNA sequence（GenBank accession number KF279661）, named ScChiⅦ1, was isolated from smut resistant sugarcane genotype Yacheng05-179. The results showed that ScChiⅦ1 was 907bp long, encoding a polypeptide of 299 amino acids. The deduced protein belonged to glycoside hydrolase family 19 and was predicted to be an extracellular chitinase. Also, there was a similar conservative structure domain of the gene of lysozyme like family, which indicating that ScChiVUl maybe has the lysozyme activity. Clustering analysis showed high homology was found among the ScChiⅦ land some Class Ⅶ chitinase genes from other plants. Genome sequence （GenBank accession number KF279662）analysis revealed that ScChiⅦ1 contained two introns which were 615 bp and 86 bp, respectively. Based on the quantitative real-time PCR analysis, the different expression patterns of ScChiⅦ1 in smut resistant genotype Yacheng05-179 and in susceptible one Liucheng03-182 were found in sugarcane buds challenged with Sporisorium scitamineum. It suggested that ScChiⅦ1 might play a role in response to the stress of S. scitamineum. When GAPDH was used as internal control, the relative expression of ScChiⅦ1 gene in buds was significantly higher than that in roots, stems, leaves and epidermis. These results could provide the foundation for further study the function of sugarcane chitinase genes and the mechanism of interaction in sugarcane-S, scitamineum biosystem.

关 键 词：甘蔗 几丁质酶 克隆 生物信息学分析 黑穗病菌 定量表达 

分 类 号：S566.1[农业科学—作物学] Q943.2[生物学—植物学]