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NlMet基因表达及其对褐飞虱繁殖力的影响

发表时间:2017-11-15  浏览量:984  下载量:123
全部作者: 魏志霜,陈丽娥,张文庆
作者单位: 中山大学生命科学学院
摘 要: 克隆褐飞虱保幼激素受体基因(NlMet),分析该基因在褐飞虱中的表达模式,并探究其对褐飞虱繁殖力的影响。提取褐飞虱的总RNA,利用反转录PCR(reverse transcription PCR,RT-PCR)及RACE(rapid-amplification of cDNA ends)法克隆得到NlMet的全长序列;利用实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)分析NlMet在褐飞虱中的龄期表达和组织分布情况,利用RNAi技术研究NlMet对褐飞虱繁殖力的影响。结果表明,褐飞虱NlMet基因全长2 895 bp,编码646个氨基酸。NlMet在若虫阶段的表达量明显低于成虫阶段的表达量,成虫7~11 d时达到表达高峰。组织表达分析表明,NlMet在脂肪体中的表达量最高。RNAi NlMet之后,导致卵黄原蛋白基因(Vg)、卵黄原蛋白受体基因(VgR)等繁殖力相关基因的表达量下调,卵巢中成熟卵数量显著减少,产卵量和后代若虫数也显著减少。研究表明,NlMet可通过调控繁殖力相关基因的表达影响褐飞虱的繁殖力。
关 键 词: 植物保护学;褐飞虱;NlMet;基因克隆;RNAi;繁殖力
Title: Gene expression and effects of methoprene-tolerant on the fecundity of the brown planthopper
Author: WEI Zhishuang, CHEN Li’e, ZHANG Wenqing
Organization: School of Life Sciences, Sun Yat-sen University
Abstract: The full-length cDNA of NlMet gene was successfully cloned from the brown planthopper, then its expression and effects on the fecundity of the brown planthopper were analyzed and explored. The full-length sequence of NlMet was cloned by using reverse transcription PCR (RT-PCR) and rapid-amplification of cDNA ends (RACE) method. Quantitative real-time PCR (qRT-PCR) and RNAi were carried out to compare the expression of NlMet in different stages and tissues, and to study the effect of NlMet on the fecundity of the brown planthopper, respectively. Sequence analysis shows that the cDNA of NlMet gene has a full-length of 2 895 bp which encodes a 646-amino acid protein. The results showed that mRNA expression level of NlMet in larva was much lower than in the counterpart of adult brown planthopper, which reached to a peak at day 7 to 11. qRT-PCR also showed that NlMet was distributed in cuticle, midgut, head, ovary and fat body, and was exhibited the highest expression in fat body. After the RNAi of NlMet gene was conducted, the mRNA expression of fecundity-related genes, such as vitellogenin (Vg) and vitellogenin receptor (VgR), was decreased. After injected dsNlMet, the number of mature eggs in ovary dropped significantly. In addition, number of eggs and offspring nymph per female also reduced significantly compared with the control. Finally, these results suggested that NlMet might regulate the fecundity of brown planthoppers by affecting some fecundity-related genes.
Key words: plant protection; brown planthopper; NlMet; gene clone; RNAi; fecundity
发表期数: 2017年11月第21期
引用格式: 魏志霜,陈丽娥,张文庆. NlMet基因表达及其对褐飞虱繁殖力的影响[J]. 中国科技论文在线精品论文,2017,10(21):2389-2397.
 
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