褐飞虱核糖体小亚基蛋白S8基因NlRPS8的克隆及其表达分析  

Cloning and expression analysis of ribosomal protein S8 gene,NlRPS8 in the brown planthopper(Nilaparvata lugens)

作　　者：朱家骏[1] 郝培应[1] 陆潮峰 冯娅琳 俞晓平[1] 

机构地区：[1]中国计量学院生命科学学院,浙江省生物计量及检验检疫技术重点实验室,杭州310018

出　　处：《应用昆虫学报》2016年第5期1036-1044,共9页Chinese Journal of Applied Entomology

基　　金：国家自然科学基金项目(31171860,30900944);国家重点基础研究发展计划(“973”计划)项目(2012CB114100,2011CB111602);浙江省公益性技术应用研究计划项目(2012C22041)

摘　　要：【目的】核糖体蛋白在生物体内具有重要作用,本研究旨在探明核糖体蛋白S8在褐飞虱Nilaparvata lugens生长发育过程中的表达规律和功能。【方法】本文根据褐飞虱基因表达谱提供的差异表达基因信息及转录组提供的基因核心序列,结合褐飞虱基因组比对分析,对褐飞虱核糖体S8基因进行了预测,并通过RT-PCR获得了褐飞虱核糖体小亚基蛋白S8的全长c DNA序列,命名为NlRPS8（Gen Bank登录号：KU341337）。【结果】NlRPS8基因全长627 bp,编码208个氨基酸。进化分析表明,褐飞虱与桑粉介壳虫Maconellicoccus hirsutus亲缘关系最为接近。应用荧光定量PCR分析了基因NlRPS8的表达规律,结果表明,NlRPS8基因在褐飞虱怀卵雌虫中表达量最高,而在雄成虫、初羽化雌成虫与1~5龄若虫表达量相对较低;在褐飞虱体内,NlRPS8基因在卵巢中表达量最高,中肠次之,在其他部位表达量较低。在抗性品种ASD7和RHT上,NlRPS8基因表达量分别为感性品种TN1上的1.99倍和2.14倍。NlRPS8基因在经过饥饿处理1 d的褐飞虱组中表达量为正常组的1.6倍。【结论】研究结果显示NlRPS8基因在褐飞虱生长、繁殖中发挥作用,为进一步研究NlRPS8基因在褐飞虱生长发育和对抗性品种适应中的功能提供了线索。[Objectives] Ribosomal proteins play very important roles in the growth and development of organisms. This study aimed to determine the expression pattern and function of a ribosomal protein S8 gene, NlRPS8（Gen Bank No. KU341337） in the rice brown planthopper, Nilaparvata lugens. [Methods] The full length c DNA of NlRPS8 was amplified from N. lugens by RT-PCR and rapid-amplification of c DNA ends using RNA-seq analysis of transcriptome and gene expression profile information. [Results] The results show that NlRPS8 has an open reading frame of 627 bp encoding 208 amino acid residues. A phylogenetic tree based on variation in the NlRPS8 gene shows that N. lugens is most closely related to Maconellicoccus hirsutus. q RT-RCR analysis revealed that the m RNA level was highest in gravid females but relatively low in adult males, newly eclosed female adults, and nymphs. q RT-RCR analysis of the head, thorax, fat body, ovaries, and mid gut, revealed that NlRPS8 was most highly expressed in the ovaries. The relative expression levels of NlRPS8 in the resistant rice varieties ASD7 and RHT were 1.99 and 2.14 times, respectively, that in the susceptible rice variety TN1. The relative expression of NlRPS8 in a treatment group provided with water only was 1.6 times that of a group fed TN1 rice. [Conclusion] This study provides new clues for further research on the function of RPS8 in N. lugens development and its adaptation to resistant rice varieties.

关 键 词：褐飞虱 NlRPS8基因 RT-qRCR 致害性变异 

分 类 号：S435.112.3[农业科学—农业昆虫与害虫防治]