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园艺学报 ›› 2014, Vol. 41 ›› Issue (6): 1218-1226.

• 研究报告 • 上一篇    下一篇

岷江百合中黄瓜花叶病毒诱导的LrPR10的克隆及表达分析

张响玲,张延龙*,牛立新,孙道阳,梁振旭   

  1. (西北农林科技大学林学院,陕西杨凌 712100)
  • 收稿日期:2013-11-20 出版日期:2014-06-25 发布日期:2014-06-25

Cloning and Expression Analysis of LrPR10 Gene from Lilium regale Induced by Cucumber mosaic virus

ZHANG Xiang-ling,ZHANG Yan-long*,NIU Li-xin,SUN Dao-yang,and LIANG Zhen-xu   

  1. (College of Forestry,Northwest A & F University,Yangling,Shaanxi 712100,China)
  • Received:2013-11-20 Online:2014-06-25 Published:2014-06-25

摘要: 为了研究黄瓜花叶病毒(CMV)诱导的岷江百合(Lilium regale)病程相关蛋白PR10基因在抗病毒防御反应中的作用,对岷江百合叶片接种CMV,采用RACE技术获得岷江百合LrPR10的全长cDNA序列,并对其进行生物信息学分析;利用实时荧光定量PCR对LrPR10在各器官特异性以及CMV和水杨酸(SA)处理后的表达模式进行了测定分析。结果表明:LrPR10全长756 bp,可编码由157个氨基酸组成的蛋白质,该蛋白具有病程相关蛋白典型的Bet_v1_like保守结构域;LrPR10在岷江百合鳞茎中相对表达量最高,在嫩叶中最低;该基因可以被CMV和SA诱导上调表达,岷江百合、卷丹(L. lancifolium)和宜昌百合(L. leucanthum)在CMV接种处理后LrPR10的相对表达量分别在4 d、4 d和1 d达到最大值,分别为处理前的58倍、27倍和292倍,在SA处理后LrPR10的相对表达量都在8 h达到最大值,分别为处理前的34倍、8倍和38倍。以上结果表明LrPR10在岷江百合抗黄瓜花叶病毒防御反应过程中发挥作用。

关键词: 岷江百合, 黄瓜花叶病毒, LrPR10, RACE, 表达分析

Abstract: This study aimed to clone PR10 gene which was up-regulated in the leaves of Lilium regale infected by Cucumber mosaic virus(CMV)and examine the role of PR10 gene in defense response against CMV.The full length of LrPR10 was amplified by RACE and analyzed with bioinformatics tools. Real-time PCR was performed to check the transcript levels of LrPR10 in different organs of L. regale,CMV-inoculated and salicylic acid(SA)–treated leaves. The results showed that the length of complete cDNA of LrPR10 was 756 bp,which encoded 157 amino acids consisting of a Bet_v1_like conserved domain of pathogenesis-related proteins. The transcript level of LrPR10 was the highest in bulbs,whereas the lowest in tender leaves. LrPR10 was significantly induced by CMV inoculation and SA treatment. Transcript level of LrPR10 reached a peak at 4 d,4 d and 1d in the CMV-inoculated leaves of L. regale L. lancifolium and L. leucanthum respectively,while the maximum relatively expression values in CMV-inoculated samples were about 58,27 and 292 times higher than the uninoculated ones in these three species. After SA treatment,LrPR10 peaked at 8 h in L. regaleL. lancifolium and L. leucanthum and the maximum relatively expression values in treated samples were about 34,8 and 38 times higher than the untreated ones in three species. We hypothesize that LrPR10 from L. regale probably plays an essential role in plant defenseagainst CMV.

Key words: Lilium regale, Cucumber mosaic virus, LrPR10, RACE, expression analysis

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