禾谷缢管蚜水通道蛋白基因RpAQP1的克隆、分子特性和表达分析  

Molecular cloning,characterization and expression analysis of aquaporin gene RpAQP1 in bird cherry-oat aphid,Rhopalosiphum padi

作　　者：左亚运 李晓叶[1] 李玉婷[1] 王康[1] 乔宪凤[1] 陈茂华[1] 

机构地区：[1]西北农林科技大学植物保护学院,农业部西北黄土高原作物有害生物综合治理重点实验室,陕西杨凌712100

出　　处：《植物保护学报》2014年第6期657-664,共8页Journal of Plant Protection

基　　金：国家自然科学基金(31272036;31471766);教育部高等学校博士学科点专项科研基金(20110204110001)

摘　　要：为探究禾谷缢管蚜Rhopalosiphum padi(Linnaeus)水通道蛋白基因Rp AQP1的序列特征及其在不同龄期的表达,利用RT-PCR和RACE技术克隆了Rp AQP1基因的c DNA全序列,采用生物信息学软件分析了Rp AQP1编码蛋白的特性,利用qRT-PCR分析了Rp AQP1在不同龄期的表达量。结果显示:禾谷缢管蚜水通道蛋白基因Rp AQP1的c DNA全长为1 216 bp,其750 bp的开放阅读框编码250个氨基酸,蛋白分子量为27.36 k D。Rp AQP1属于水通道蛋白亚家族DRIP(果蝇内嵌蛋白Drosophila integral protein)的一员,具有6个跨膜区,2个保守的NPA结构单元和1个压缩区域Ar/R;qRT-PCR结果显示,Rp AQP1在整个发育历期均有表达,其在2龄若蚜中表达水平最高,是成蚜表达量的1.432倍;在4龄若蚜中表达量最低,为成蚜表达量的0.444倍,显著低于其它龄期,而其余各龄期Rp AQP1表达量无显著差异。To amplify the sequence of aquaporin gene Rp AQP1 and analyze the expression pattern of Rp AQP1 at different developmental stages of bird cherry-oat aphid,Rhopalosiphum padi,the reverse transcription PCR( RT-PCR) and rapid-amplification of c DNA ends( RACE) were used to amplify the full c DNA of Rp AQP1 gene; real time PCR( qRT-PCR) method was used to analyze the expression profile of the gene at different stages of the pest. The results showed that the full-length Rp AQP1 c DNA was 1 216 bp,which included an open reading frame of 750 bp encoding a protein of 250 amino acids with a predicted molecular mass of 27. 36 k D. Rp AQP1 was a member of the DRIP protein family.Rp AQP1 exhibited a typical structure with six membrane-spanning domains,two highly conserved AsnPro-Ala( NPA) motifs,and the Ar / R selectivity regions of AQP1 family. qRT-PCR analysis showed that this gene was expressed at all developmental stages of R. padi. The highest expression level of Rp AQP1 was found in the 2nd instar,which was 1. 432 times of the expression level in adult aphid. The lowest expression level of the gene was found in the 4th instar,0. 444 times of the expression level in adultaphid and significantly lower than those in all other developmental stages. There was no significant difference between the Rp AQP1 expression levels in other instars.

关 键 词：禾谷缢管蚜 水通道蛋白 基因克隆 分子特性 表达分析 

分 类 号：S433.89[农业科学—农业昆虫与害虫防治]