CAJ | 학술논문

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 HaPR1的 cDNA 全长序列,并进行了表达模式和功能分析。结果表明,该基因 cDNA 全长开放阅读框为489 bp,编码162个氨基酸,分子量为17.52 kD,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, GenBank 登录号为 KR071874。经比较 HaPR1与多种物种 PR1高度同源。实时荧光定量 PCR 检测结果表明, HaPR1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 HaPR1具有抗核盘菌的功能。
병정상관단백(pathogenesis-related proteins)경상피용작식물항병반응적분자표기。본문재핵반균유도향일규전록조문고적기출상극륭1개병정상관단백1기인 HaPR1적 cDNA 전장서렬,병진행료표체모식화공능분석。결과표명,해기인 cDNA 전장개방열독광위489 bp,편마162개안기산,분자량위17.52 kD,등전점위8.19,구유6개보수반광안산,4개보수적 allergen V5/Tpx-1결구역, GenBank 등록호위 KR071874。경비교 HaPR1여다충물충 PR1고도동원。실시형광정량 PCR 검측결과표명, HaPR1상대표체량재향일규협중최고,근중기차,경중최저。간한、염、초산、핵반균급기대사물균가현저유도기표체。이용농간균개도법장해기인도입연초,제고료전기인주계대핵반균적항성。대항성주계연초방어매활성급병이철함량측정발현전기인연초협편재핵반균협박하현저제고료 SOD、POD화 CAT 활성,강저료 MDA 함량。초보추단 HaPR1구유항핵반균적공능。
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 of HaPR1 (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 con-served 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 of transgenic 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.