马铃薯腐烂茎线虫类毒液过敏原蛋白基因(Dd-vap-1)的克隆与表达定位分析  

Cloning and Localization Analysis of a New Venom Allergen-like Protein Gene(Dd-vap-1) in Ditylenchus destructor

作　　者：周采文[1,2] 胡先奇[2] 彭德良[1] 彭焕[1] 龙海波[1] 黄文坤[1] 

机构地区：[1]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实验室,北京100193 [2]云南农业大学农业生物多样性应用技术国家工程研究中心,昆明650201

出　　处：《农业生物技术学报》2013年第3期299-305,共7页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(No.31071668)

摘　　要：植物寄生线虫食道腺分泌蛋白在寄生致病过程中起到至关重要的作用。用RT-PCR结合RACE的方法从马铃薯腐烂茎线虫(Ditylenchus destructor)中克隆了一个类毒液过敏原蛋白新基因Dd-vap-1。Dd-vap-1cDNA全长1346bp,开放阅读框的长度为1209bp,推导编码一个含有402个氨基酸的蛋白序列(GenBank登录号JQ341057)。推导蛋白Dd-VAP-1预测分子量为45.3kD,等电点为6.68,序列比对分析表明,Dd-VAP-1为富含半胱氨酸分泌蛋白家族成员。原位杂交结果显示,Dd-vap-1在马铃薯腐烂茎线虫的亚腹食道腺细胞特异表达,推测该基因在马铃薯腐烂茎线虫侵染甘薯的早期阶段起重要的作用。Secretory proteins in the oesophageal gland cells of plant-parasitic nematodes have key roles in nematodes parasitism of plant. A new venom allergen-like protein gene （Dd-vap-1） was cloned from Ditylenchus destructor by RT-PCR and RACE. The full length cDNA of Dd-vap-1 was 1 346 bp. The cDNA sequence consisted of a 1 209 bp open reading frame encoding 402 amino acid residues that were franked by a 69 bp 5＇-untranslated region and a 68 bp 3＇-untranslated region （GenBank accession No. JQ341057）. The theoretical molecular weight and isoelectric point of the putative protein were 45.3 kD and 6.68, respectively. The sequencing analysis showed that this gene was classified as a member of venom allergen-like protein. In situ hybridization analysis showed that the transcript of Dd-vap-1 accumulated exclusively within the subventral oesophageal cell of D. destructor. The result suggested that Dd-vop-1 may play an important role in the early stage when infecting host plants.

关 键 词：马铃薯腐烂茎线虫 类毒液过敏原蛋白基因 原位杂交 RACE 

分 类 号：S432.45[农业科学—植物病理学]