【摘要】 为探究甘蔗PROTEOLYSIS 1(PRT1)基因与黑穗病菌的互作关系，以甘蔗割手密种和栽培品种为研究对象，通过RT-PCR技术对ScPRT1(GenBank登录号：MT747433)基因进行克隆，生物信息学分析、定量PCR、亚细胞定位和基因共表达网络分析。生物信息学分析表明，该基因的cDNA全长为1 621 bp,包含一个长度为1 260 bp,编码419个氨基酸的完整开放读码框；其编码蛋白的分子量为46.56 ku,为酸性、不稳定的亲水蛋白，无信号肽，具有核定位信号；该蛋白包含2个RING结构域和1个ZZ结构域；ScPRT1蛋白的高级结构元件多为无规则卷曲。定量PCR结果显示，ScPRT1在皮、叶片、芽中的表达量相差不大，在蔗髓中表达量最高。同时该基因的表达受脱落酸胁迫后显著上调；受黑穗病菌胁迫后，在甘蔗抗黑穗病品种中上调表达，在感黑穗病品种中下调表达，都达到了显著水平。亚细胞定位结果揭示，ScPRT1定位于细胞核。寄主甘蔗和病原黑穗病菌的基因共表达网络中，与甘蔗ScPRT1共表达的黑穗病菌基因中，有3个为N端具有芳香族氨基酸残基的黑穗病菌效应蛋白，这暗示它们之间存在直接的相互作用。以上结果表明，ScPRT1在甘蔗激素信号传导以及响应黑穗病菌侵染过程中发挥重要作用。更多还原

【Abstract】 In order to explore the interaction between ScPRT1 and Sporisorium scitamineum genes in sugarcane, this study used Saccharum spontaneum and Saccharum as research objects.The ScPRT1(GenBank Accession Number: MT747433)gene was cloned by RT-PCR and was analyzed by bioinformatics, quantitative Real-time PCR and subcellular localization.Bioinformatics analysis showed that the full-length cDNA of the ScPRT1 gene was 1 621 bp, including a complete open reading frame with 1 260 bp in length and encodes 419 amino acids.The ScPRT1 protein with two RING domains and one ZZ domain, was 46.56 ku in molecular weight and was an acidic and unstable hydrophilic protein with nuclear localization signal and no signal peptide.The higher structural elements of protein were mainly random coil.The results of qRT-PCR analysis showed that ScPRT1 was higher expressed in stem pith than in bud, epidermis and leaf.And ScPRT1 was significantly up-regulated under abscisic acid(ABA)stress and in the smut-resistant variety but significantly down-regulated in the smut-susceptible variety under Sporisorium scitamineum stress.The result of subcellular localization showed that ScPRT1 was localized in the nucleus.In the co-expression network between ScPRT1 and Sporisorium scitamineum genes, three genes that encoded effectors with aromatic amino acid residues at the N-terminal were found co-expressing with ScPRT1,and their proteins may interact directly.It is speculated that ScPRT1 plays an important role in sugarcane hormone signal transduction and response to smut infection.更多还原