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褐飞虱Nilaparvata lugens(Stål)生殖力的蛋白质组学和转录组学分析。

Proteomic and transcriptomic analyses of fecundity in the brown planthopper Nilaparvata lugens (Stål).

机构信息

State Key Laboratory of Biocontrol and School of Life Sciences, Sun Yat-Sen University , No. 135 Xingang West Road, Guangzhou 510275, China.

出版信息

J Proteome Res. 2013 Nov 1;12(11):5199-212. doi: 10.1021/pr400561c. Epub 2013 Oct 10.

DOI:10.1021/pr400561c
PMID:24083549
Abstract

As an r-strategy insect species, the brown planthopper (BPH) Nilaparvata lugens (Stål) is a serious pest of rice crops in the temperate and tropical regions of Asia and Australia, which may be due to its robust fecundity. Here we combined 2-DE comparative proteomic and RNA-seq transcriptomic analyses to identify fecundity-related proteins and genes. Using high- and low-fecundity populations as sample groups, a total of 54 and 75 proteins were significantly altered in the third and sixth day brachypterous female stages, respectively, and 39 and 54 of these proteins were identified by MALDI-TOF/TOF MS. In addition, 71,966 unigenes were quantified by Illumina sequencing. On the basis of the transcriptomic analysis, 7408 and 1639 unigenes demonstrated higher expression levels in the high-fecundity population in the second day brachypterous female adults and the second day fifth instar nymphs, respectively, and 411 unigenes were up-regulated in both groups. Of these dozens of proteins and thousands of unigenes, five were differentially expressed at both the protein and mRNA levels at all four time points, suggesting that these genes may regulate fecundity. Glutamine synthetase (GS) was chosen for further functional studies. RNAi knockdown of the GS gene reduced the fecundity of N. lugens by 64.6%, disrupted ovary development, and inhibited vitellogenin (Vg) expression. Our results show that a combination of proteomic and transcriptomic analyses provided five candidate proteins and genes for further study. The knowledge gained from this study may lead to a more fundamental understanding of the fecundity of this important agricultural insect pest.

摘要

作为一种 r 策略昆虫物种,褐飞虱(BPH)Nilaparvata lugens(Stål)是亚洲和澳大利亚温带和热带地区水稻作物的严重害虫,这可能是由于其强大的繁殖力。在这里,我们结合 2-DE 比较蛋白质组学和 RNA-seq 转录组学分析来鉴定与繁殖力相关的蛋白质和基因。使用高繁殖力和低繁殖力种群作为样本组,在第三和第六天短翅雌虫阶段,分别有 54 和 75 种蛋白质显著改变,其中 39 和 54 种蛋白质通过 MALDI-TOF/TOF MS 鉴定。此外,通过 Illumina 测序定量了 71966 个基因。基于转录组分析,在高繁殖力种群中,有 7408 和 1639 个基因在第二天短翅雌成虫和第二天第五龄若虫中表达水平较高,而在两组中均有 411 个基因上调。在这些数十种蛋白质和数千个基因中,有 5 种在所有四个时间点在蛋白质和 mRNA 水平上均有差异表达,表明这些基因可能调节繁殖力。谷氨酰胺合成酶(GS)被选为进一步的功能研究。GS 基因的 RNAi 敲低使 N. lugens 的繁殖力降低了 64.6%,破坏了卵巢发育,并抑制了卵黄原蛋白(Vg)的表达。我们的研究结果表明,蛋白质组学和转录组学分析的结合为进一步的研究提供了五个候选蛋白质和基因。这项研究获得的知识可能会导致对这种重要农业害虫繁殖力的更深入理解。

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