割手密2C型蛋白磷酸酶ScPP2C基因的克隆与表达分析

Cloning and Expression Analysis of the Sc PP2C Gene in S. spontaneum

割手密由于具有抗病、抗虫、抗逆等潜在的有利基因,历来被用于甘蔗的抗性遗传改良。在割手密中克隆编码2C型蛋白磷酸酶(protein phosphatase 2C, PP2C)的基因,并分析其在干旱胁迫下的差异表达。本研究利用RT-PCR方法从割手密叶片中克隆得到PP2C基因的CDS全长序列,利用生物信息学在线软件对PP2C蛋白的理化性质、蛋白结构进行预测分析,并采用实时荧光定量的方法分析该基因在不同干旱处理下的差异表达。结果显示从割手密中克隆到一个全长951 bp编码316个氨基酸的2C型蛋白磷酸酶基因,命名为ScPP2C,GenBank登录号为MG322120。荧光定量分析表明,随着干旱胁迫时间的延长,其表达量呈先上调后下调的表达模式,说明该基因是干旱胁迫诱导型基因。本研究通过挖掘甘蔗野生种割手密中的抗旱新基因ScPP2C,为甘蔗野生种质资源开发利用、转基因抗旱甘蔗新品种的选育提供了参考依据。

S. spontaneum L. has always been an important germplasm resource for sugarcane genetic improvement due to its potential beneficial genes such as disease resistance, insect resistance and stress resistance, etc. In this study, a PP2 C gene designated as ScPP2 C(GenBank Accession: MG322120), which including an ORF of 951 bp coding sequence and coding 316 amino acids, was amplified from Saccharum spontaneum L., Realtime-PCR analysis revealed that the expression level of ScPP2 C was up-down-regulated under different drought conditions,indicating that the gene is a drought stress-inducible gene. In this study, a novel gene of drought-resistance was found in S. spontaneum L., which was to develope the excellent resources of wild species, as well as to explore the drought resistance of these resources, so as to provide the basis for the selection of the parents of the fine and drought resistance in the future.