《植物生理学报》 2012, 48(1): 57-62

葡萄病程相关蛋白1基因的克隆和表达分析

侯丽霞, 高超, 车永梅, 赵方贵, 刘新*

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

通信作者：刘新；E-mail: liuxin6080@yahoo.com.cn；Tel: 0532-8030224

摘　要：

以葡萄品种‘左优红’组培苗叶片为材料, 利用同源克隆法获得其病程相关蛋白1基因VvPR1的cDNA全长序列。扩增片段大小为486 bp, 编码161个氨基酸, 分子量17.5 kDa, 等电点PI=8.69, 含有6个保守半胱氨酸, 4个allergen V5/Tpx-1 related保守结构域。VvPR1与多种植物PR1高度同源。实时定量PCR检测结果表明VvPR1在葡萄叶片中相对表达量最高; 霜霉病菌、低温、盐和干旱胁迫均可显著诱导其表达; 水杨酸、脱落酸、茉莉酸、一氧化氮、过氧化氢和硫化氢等亦可诱导其大量表达, 据此推测, VvPR1参与了多种生物胁迫和非生物胁迫过程。

关键词：葡萄; 病程相关蛋白1; 基因克隆; 表达分析

收稿：2011-07-27   修定：2011-10-14

资助：农业部‘948’项目(2006-G26)和青岛市自然基金(10-3-4-5-5-jch)。

Gene Cloning and Expression Analysis of Pathogenesis-Related Protein 1 in Vitis vinifera L.

HOU Li-Xia, GAO Chao, CHE Yong-Mei, ZHAO Fang-Gui, LIU Xin*

College of Life Sciences, Qingdao Agricultural University, Key Lab of Plant Biotechnology in Universities of Shangdong Province, Qingdao, Shandong 266109, China

Corresponding author: LIU Xin; E-mail: liuxin6080@yahoo.com.cn; Tel: 0532-8030224

Abstract:

Using homology cloning method, the full-length cDNA of pathogenesis-related protein 1 (PR1) named VvPR-1 was cloned from leaves of Vitis vinifera cultivar ‘Zuoyouhong’ tissue culture seedling. Bioinformatic analysis indicated that VvPR-1 consisted of 486 nucleotides encoding 161 amino acid with molecular weight 17.5 kDa, isoelectric point 8.69, VvPR-1 possessed six conserved cysteine and four conserved allergen V5/Tpx-1 related domain. The VvPR-1 was highiy homologous to PR1 in other plants. Real-time PCR analysis showed that expression level of VvPR-1 was the highest in leaves, VvPR1 was significantly induced by Plasmopara viticola inoculation, salt stress and osmotic stress, signaling molecules such as salicylic acid (SA), jasmonate (JA), abscisic acid (ABA) as well as nitric oxide (NO), hydrogen peroxide (H2O2), hydrogen sulfide (H2S) also had inductive effects on VvPR-1 expression. It suggested that VvPR1 was concerned with various biotic stresses and abiotic stresses.

Key words: Vitis vinifera; pathogenesis-related protein 1; gene clone; expression analysis