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苗期水稻响应褐飞虱取食的基因差异表达分析
Change in the gene expression of seedling stage rice in response to feeding by the brown planthopper Nilaparvata lugens (St?l)(Hemiptera: Delphacidae)
曹征鸿;贺 康;徐 乐;汤沈杨;王亚琴;李 飞
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DOI:10.7679/j.issn.2095-1353.2020.093
作者单位:浙江大学昆虫科学研究所;水稻生物学国家重点实验室;浙江大学生物技术研究所
中文关键词:水稻;飞虱;转录组;分子响应
英文关键词:rice; Nilaparvata lugens; rranscriptome analysis; molecular response
中文摘要:
【目的】 褐飞虱Nilaparvata lugens (Stål) 是水稻的重要害虫之一,主要刺吸水稻韧皮部汁液为害,对水稻生产造成严重的产量和经济损失。为研究水稻响应褐飞虱取食的分子机制,对褐飞虱取食6 h后的苗期水稻进行转录组测序及基因差异表达分析。【方法】 采用illumina二代测序技术获得褐飞虱取食前后水稻组织的转录组数据,利用RSEM软件进行基因表达定量和DEseq2进行差异表达分析;从差异表达基因中随机选取20个基因采用荧光定量PCR技术进行验证;采用GeneMerge软件对差异表达基因进行KEGG和GO富集分析。【结果】 褐飞虱取食后,水稻转录组中的1 104个基因出现了差异表达,其中435个基因表达上调,669个基因表达下调。荧光定量PCR结果显示,20个差异表达基因中18个基因的表达变化趋势和测序结果一致,证明了转录组分析结果可靠。GO和KEGG富集分析表明,表达上调基因主要与水稻氧化应激、海藻糖合成及次生化合物代谢有关,显著富集在14个KEGG通路和30个GO功能分类中;而表达下调基因主要参与水稻纤维素、蛋白质及脂肪酸合成过程,显著富集在29个KEGG通路和26个GO功能分类。在差异表达基因中,分别有61个转录因子和13个水杨酸和茉莉酸信号通路相关基因。【结论】 褐飞虱取食激发了水稻的应激反应和保护机制,同时还降低了营养合成的过程,是飞虱为害造成水稻减产的原因之一。本研究初步揭示了苗期水稻响应褐飞虱取食的差异表达基因,为研究水稻-褐飞虱互作机制以及褐飞虱抗性水稻品种培育提供了参考和依据。
英文摘要:
[Objectives]  To investigate the molecular mechanisms used by plants in response to attack by the brown planthopper BPH; [Nilaparvata lugens (Stål)], a piercing-sucking insect that is now one of the most globally important rice pests. [Methods]  Transcriptome data of seedling stage rice, with, or without, BPH were obtained using Illumina next-generation sequencing technology. Gene expression levels were obtained using RSEM software and DEseq2 used to analyze differential gene expression. Twenty differentially expressed genes were randomly selected for verification with fluorescent quantitative real-time PCR (qPCR) and GeneMerge software was used to analyze KEGG and GO enrichment of differentially expressed genes. [Results]  A total of 1 104 differentially expressed genes were identified in rice plants infested by BPH, of which 435 were up-regulated and 669 were down-regulated. Twenty differentially expressed genes were selected for validation with qPCR, among which 18 were consistent with RNA-Seq estimation thereby confirming the reliability of transcriptome analysis. GO and KEGG enrichment analysis show that the up-regulated genes were enriched into 14 KEGG pathways and 30 GO terms, that are mainly involved in oxidative stress, trehalose synthesis and secondary metabolism, whereas the down-regulated genes were enriched into 29 KEGG pathways and 26 GO terms that are mainly involved in the synthesis of cellulose, protein and fatty acids. In addition, 61 transcription factors and 13 genes related to the salicylic acid and jasmonic acid pathways were enriched in the differentially expressed genes. [Conclusion]  Feeding by the BPH stimulates the stress and defense responses of rice plants, and also reduces nutrient synthesis, which suggests a possible impact of BPH on rice yield. These results reveal changes in gene expression in rice plants in response to feeding by BPH, the possible impact of BPH on rice yield, and also provide theorical support for developing rice cultivars resistant to BPH.
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