摘要
病程相关蛋白(pathogenesis-related proteins)经常被用作植物抗病反应的分子标记。本文在核盘菌诱导向日葵转录组文库的基础上克隆1个病程相关蛋白1基因Ha PR1的c DNA全长序列,并进行了表达模式和功能分析。结果表明,该基因c DNA全长开放阅读框为489 bp,编码162个氨基酸,分子量为17.52 k D,等电点为8.19,具有6个保守半胱氨酸,4个保守的allergen V5/Tpx-1结构域,Gen Bank登录号为KR071874。经比较Ha PR1与多种物种PR1高度同源。实时荧光定量PCR检测结果表明,Ha PR1相对表达量在向日葵叶中最高,根中其次,茎中最低。干旱、盐、草酸、核盘菌及其代谢物均可显著诱导其表达。利用农杆菌介导法将该基因导入烟草,提高了转基因株系对核盘菌的抗性。对抗性株系烟草防御酶活性及丙二醛含量测定发现转基因烟草叶片在核盘菌胁迫下显著提高了SOD、POD和CAT活性,降低了MDA含量。初步推断Ha PR1具有抗核盘菌的功能。
Pathogenesis-related proteins are commonly used as markers of plant defense responses. The full-length cDNA of pathogenesis-related protein 1 （PR1） named HaPR1 in Helianthus annuus was cloned based on the transcriptome of H. annuus induced by Sclerotinia sclerotiorum, and its expression model and function were analyzed in this study. Sequence analysis showed that the cDNA ofHaPR1 （GenBank accession No. KR071874） contained a 489 bp ORF encoding a protein of 162 amino acids residues with the molecular mass of 17.52 kD and theoretical pI of 8.19, HaPR1 possessed six conserved cysteine and four conserved allergen V5/Tpx-1 related domain. The HaPR1 was highly homologous with PR1 in other species. Real-time PCR analysis showed that expression level of HaPR1 was the highest in leaf, and was significantly induced by drought, salt stress, oxalic acid, S. sclerotiorum and its metabolites. Then the HaPR1 gene was transformed into tobacco by Agrobacterium tumefaciens to further verify its function. The results showed that the expression of HaPR1 improved the resistance oftransgenic lines, and significantly increased SOD, POD, and CAT activities and reduced the content of MDA. It suggested that HaPR1 has a function of resistance to S. sclerotiorum.
出处
《作物学报》
CAS
CSCD
北大核心
2015年第12期1819-1827,共9页
Acta Agronomica Sinica
基金
国家现代农业产业技术体系建设专项(CARS-16)
黑龙江省农业科技创新工程项目(QN015)资助~~
关键词
向日葵
病程相关蛋白1
基因克隆
功能分析
Helianthus annuus
Pathogenesis-related protein 1
Gene clone
Function analysis
