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烟粉虱MED隐种蜕皮激素受体基因cDNA克隆、转录与组织表达

引用本文：杨春红,龙楚云,郭建洋,程登发,万方浩.烟粉虱MED隐种蜕皮激素受体基因cDNA克隆、转录与组织表达.植物保护学报,2016,43(1):40-47

DOI：10.13802/j.cnki.zwbhxb.2016.01.006

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作者	单位	E-mail
杨春红	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
龙楚云	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
郭建洋	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
程登发	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	dfcheng@ippcaas.cn
万方浩	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 青岛农业大学农学与植物保护学院, 青岛 266109	wanfanghao@caas.cn

中文摘要:蜕皮激素受体(ecdysone receptor,EcR)是调控昆虫蜕皮、变态和生殖等过程中基因表达的重要调控因子.为研究EcR在烟粉虱Bemisia tabaci生长发育中的作用,利用RT-PCR和RACE等技术扩增了烟粉虱MED隐种EcR基因的cDNA全长序列,并利用实时荧光定量PCR技术检测其在烟粉虱不同发育时期相对表达量的变化.BtEcR cDNA序列全长2 844 bp,含有一个1 518 bp的开放阅读框,共编码505个氨基酸,预测蛋白分子量为56 kD,等电点为6.43,含有特殊结构功能域:DNA结合域(DBD_EcR)和配体结合域(LBD_EcR),该序列编码的蛋白质序列与其它已报道的昆虫EcR蛋白序列具有很高的相似性,命名为BtEcR(GenBank登录号: KR534774).BtEcR在MED隐种若虫、雌成虫各发育时期均有表达,若虫期在伪蛹前期达到最高值,成虫期呈现先升高后降低的趋势,且在羽化后第7天达到最高值,约为羽化第1天的23倍.研究结果为揭示BtEcR在烟粉虱整个生长发育过程中的作用提供了重要依据.

中文关键词:烟粉虱蜕皮激素受体核受体表达谱荧光定量PCR

Molecular cloning,sequence analysis and expression of ecdysone receptor gene from Bemisia tabaci Mediterranean (Hemiptera: Aleyrodidae)

Author Name	Affiliation	E-mail
Yang Chunhong	State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China
Long Chuyun	State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China
Guo Jianyang	State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China
Cheng Dengfa	State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China	dfcheng@ippcaas.cn
Wan Fanghao	State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China Qingdao Agricultural University, Qingdao 266109, Shandong Province, China	wanfanghao@caas.cn

Abstract:Ecdysone receptor (EcR) is a dominant regulator of development, metamorphosis and reproduction in insects. In order to define the function of EcR gene in Bemisia tabaci Mediterranean (MED), the full-length cDNA sequence of an EcR gene was cloned by both reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods, and the relative expression levels at different developmental stages of MED was analyzed using real-time quantitative PCR (qPCR). The complete cDNA (2 844 bp) of EcR gene in B. tabaci MED species complex contained an open reading frame (ORF) of 1 518 bp, encoding 505 amino acids, with a molecular weight of 56 kD and a theoretical isoelectric point of 6.43. This gene had two special domains: the DNA-binding domain of ecdysone receptor (DBD_EcR) and the ligand-binding domain of ecdysone receptor (LBD_EcR). Multiple sequence alignment revealed that the deduced amino acid sequence had high identity with EcR from other insect species. This cDNA was named as BtEcR (GenBank accession no. KR534774). qPCR results indicated that BtEcR transcripts were detected at all instars of nymph and developmental stages of female adults. The highest relative expression level of EcR gene was detected at pseudo-pupae stage among different instar nymphs. Peak level was detected at 7 d after eclosion, which was about 23 times the level at 1 d after eclosion. The present results provided valuable information for revealing the role of BtEcR underlying the development of whitefly species.

keywords:Bemisia tabaci ecdysone receptor nuclear receptor expression profile real-time quantitative PCR

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