烟粉虱MEAM1隐种漆酶-1基因全长cDNA克隆、序列分析与组织表达

›› 2013, Vol. 56 ›› Issue (2): 111-119.doi:

• 研究论文 • 下一篇

烟粉虱MEAM1隐种漆酶-1基因全长cDNA克隆、序列分析与组织表达

杨春红^1,2，彭露³，郭建洋²，严盈²，万方浩^2,*，王进军¹

（1. 西南大学植物保护学院，昆虫学及害虫控制工程重庆市市级重点实验室，重庆 400716； 2. 中国农业科学院植物保护研究所，植物病虫害生物学国家重点研究室，北京 100193； 3. 福建农林大学生物安全学报编辑部，福州350002）

出版日期:2013-02-20 发布日期:2013-02-20

Molecular cloning, sequence analysis and expression of a laccase-1 gene from Bemisia tabaci Middle East-Asia Minor 1 (Hemiptera: Aleyrodidae)

YANG Chun-Hong^1,2, PENG Lu³, GUO Jian-Yang², YAN Ying², WAN Fang-Hao^2,*, WANG Jin-Jun¹

(1. Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing 400716, China; 2. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China; 3. Editorial Department of Journal of Biosafety, Fujian Agriculture and Forestry University, Fuzhou 350002, China)

Online:2013-02-20 Published:2013-02-20

摘要/Abstract

摘要： 多酚氧化酶是多种刺吸式昆虫唾液中的关键酶类，可以通过干扰寄主植物正常的氧化还原反应来调控食物来源使其更有利于昆虫本身。昆虫体内的漆酶-1属于多酚氧化酶家族，被认为参与昆虫取食过程中的金属离子代谢、降解植物次生有毒物质以及免疫防御等生理活动。为研究漆酶-1基因在烟粉虱MEAM1隐种Bemisia tabaci Middle East-Asia Minor 1组织中的功能，利用RACE技术首次获得了烟粉虱MEAMI隐种漆酶-1（laccase-1，lac-1）基因的cDNA全长序列，命名为Btlac-1 ( GenBank 登录号： JQ966215）。结果表明，该基因含有一个2 733 bp的开放阅读框，编码910个氨基酸，其编码产物含有3个Cu-oxidase功能域，属于蓝多铜氧化酶家族成员。同源性比较分析表明，Btlac-1与黑尾叶蝉Nephotettix cincticeps和豌豆蚜Acyrthosiphon pisum的lac-1编码的氨基酸序列一致性达58%。Btlac-1在不同组织和发育时期的转录表达分析表明, Btlac-1在烟粉虱MEAM1隐种中肠中的转录水平最高，显著高于头胸部和腹部组织中，且在各个发育时期均有表达，在成虫期的表达水平较高。该结果为进一步明确lac-1在烟粉虱取食过程的生理作用奠定基础，也为探讨刺吸式昆虫与植物相互作用机制提供理论依据。

关键词: 烟粉虱MEAM1隐种, 漆酶-1基因, 多酚氧化酶, 表达谱, 荧光定量PCR

Abstract: Polyphenol oxidase is the predominant enzyme in saliva of piercing-sucking insects. It can interfere with the normal oxidation-reduction reaction in host plants of insects and make food source more acceptable for itself. Insect laccase-1 (lac-1) belongs to the polyphenol oxidase family, and it has been verified that it is involved in the iron homeostasis, immune defense and detoxification. But the function of lac-1 in Bemisia tabaci is not clear. To determine the function of lac-1 gene in different tissues of Bemisia tabaci Middle East-Asia Minor 1 (MEAM1), lac-1 gene in B. tabaci MEAM1 was cloned using RACE method and named Btlac-1 (GenBank accession no. JQ966215). Btlac-1 contains an open reading frame (ORF) of 2 733 bp encoding 910 amino acids, and the encoded protein has three typical Cu-oxidase domains in polyphenol oxidase family, suggesting that Btlac-1 belongs to blue copper-containing polyphenol oxidase family. Homology analysis showed that Btlac-1 shares 58% amino acid sequence identity with lac-1 genes from Nephotettix cincticeps and Acyrthosiphon pisum. Gene transcript analysis revealed that lac-1 could be expressed at every developmental stage, and the highest expression level was observed at the adult stage, suggesting that lac-1 might play an important role during the adult stage. In addition, the transcriptional level of Btlac-1 in the midgut was higher than those in cephalothorax and abdomen tissues. These results provide clues to further understanding the physiological role of lac-1 in feeding in B. tabaci, and also a basis for exploring the interaction mechanism between piercing-sucking insects and plants.

Key words: Bemisia tabaci Middle East-Asia Minor 1, lac-1, polyphenol oxidase, expression profile, Real-time quantitative PCR

杨春红, 彭露, 郭建洋, 严盈, 万方浩, 王进军. 烟粉虱MEAM1隐种漆酶-1基因全长cDNA克隆、序列分析与组织表达[J]. , 2013, 56(2): 111-119.

YANG Chun-Hong, PENG Lu, GUO Jian-Yang, YAN Ying, WAN Fang-Hao, WANG Jin-Jun. Molecular cloning, sequence analysis and expression of a laccase-1 gene from Bemisia tabaci Middle East-Asia Minor 1 (Hemiptera: Aleyrodidae)[J]. , 2013, 56(2): 111-119.

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烟粉虱MEAM1隐种漆酶-1基因全长cDNA克隆、序列分析与组织表达

Molecular cloning, sequence analysis and expression of a laccase-1 gene from Bemisia tabaci Middle East-Asia Minor 1 (Hemiptera: Aleyrodidae)

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