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园艺学报 ›› 2013, Vol. 40 ›› Issue (8): 1583-1590.

• 研究报告 • 上一篇    下一篇

牡丹病程相关蛋白1基因的克隆及表达分析

杨德翠,张玉喜,郑国生*   

  1. (青岛农业大学生命科学学院,山东省高校植物生物技术重点实验室,山东青岛 266109)
  • 收稿日期:2012-11-30 出版日期:2013-08-25 发布日期:2013-08-25

Gene Cloning and Expression Analysis of Pathogenesis-related Protein 1 of Paeonia suffruticosa

YANG De-cui,ZHANG Yu-xi,and ZHENG Guo-sheng*   

  1. (College of Life Sciences,Qingdao Agricultural University,Key Lab of Plant Biotechnology in Universities of Shangdong Province,Qingdao,Shandong 266109,China)
  • Received:2012-11-30 Online:2013-08-25 Published:2013-08-25

摘要: 以牡丹柱枝孢叶斑病病原菌Cylindrocladium canadense处理的牡丹‘鲁菏红’叶片为材料,根据已发表植物PR1的保守区域设计兼并引物,利用RT-PCR和RACE技术,获得病程相关蛋白1基因的cDNA,命名为PsPR1。该序列全长为744 bp,5′和3′非翻译区分别有42 bp和225 bp。该基因有477 bp的开放阅读框(ORF),编码158个氨基酸,成熟蛋白分子量14.8 kD,等电点(pI)6.19,具有典型的SCP_PR1_like保守结构域。通过Blast比对发现该基因与多种植物病程相关蛋白1基因具有高度同源性。通过实时荧光定量PCR检测,发现该基因在牡丹萼片中相对表达量最高,在正常生长的叶片中最少。该基因受病原菌C. canadense和信号物质SA的显著诱导,分别在处理的24 h和12 h达到最高峰,说明PsPR1可能参与了牡丹的抗病防御过程。

关键词: 牡丹, 病程相关蛋白1, 基因克隆, 表达分析

Abstract: One pair of degenerate primer was designed according to the conservative domains of the pathogenesis-related protein PR1 genes of other plants and using RT-PCR and RACE technology,a full length cDNA of pathogenesis-related protein 1,named PsPR1,was obtained from leaves of Paeonia suffruticosa‘Luhehong’,treated by Cylindrocladium canadense. Sequence analysis indicated that PsPR1 consisted of 744 bp and the 5′ UTR and 3′ UTR were 42 bp and 225 bp,respectively. The length of open reading frame(ORF)was 477 nucleotides encoding 158 amino acids with molecular weight 14.8 kD,isoelectric point 6.19. The deduced amino acids possessed SCP_PR1_like conserved domain. The PsPR1 was highly homologous to PR1,which have been isolated from other plants. Real-time PCR analysis indicated that expression level of PsPR1 was the highest in sepal,but the lowest in leaves. PsPR1 was significantly induced by C. canadense and signaling molecule salicylic acid(SA)and expression level of PsPR1 reached the highest peak in 24 h and 12 h,respectively,which showed that PsPR1 might involved in the disease defense.

Key words: Paeonia suffruticosa, pathogenesis-related protein 1, gene clone, expression analysis