大豆抗豆卷叶螟的转录组和蛋白质组关联分析

doi:10.3864/j.issn.0578-1752.2018.07.003

摘要/Abstract

摘要： 【目的】通过对大豆受豆卷叶螟幼虫胁迫下的转录组和蛋白质组结果进行联合分析，筛选出一些与大豆抗豆卷叶螟相关的候选基因，为深入认识大豆抗豆卷叶螟的分子调控机制奠定基础。【方法】以高抗材料赶泰-2-2（HR）和高感材料皖82-178（HS）为研究对象，运用RNA-Seq技术和iTRAQ技术鉴定出豆卷叶螟幼虫取食诱导0和48 h时样品间的差异表达基因（DEGs）和差异表达蛋白（DEPs），将在蛋白水平和转录水平关联到的所有可定量数据进行关联分析，计算蛋白水平和转录水平间的相关系数。【结果】蛋白质鉴定结果表明，HR48/HR0、HS48/HS0、HR0/HS0和HR48/HS48比较组中分别鉴定出236、250、213、211个DEPs；转录组鉴定结果表明，HR48/HR0、HS48/HS0、HR0/HS0和HR48/HS48比较组中分别鉴定出1 064、680、605、468个DEGs。定量关联分析结果表明，HR48/HR0、HS48/HS0、HR0/HS0和HR48/HS48比较组的相关系数r分别为0.156、0.2687、0.1149和0.035；HR48/HR0、HS48/HS0、HR0/HS0和HR48/HS48比较组中分别关联到11、9、3和4个DEPs/DEGs，其中蛋白和mRNA表达趋势相同的基因分别有11、9、2和3个，蛋白和mRNA表达趋势相反的基因分别有0、0、1和1个。生物信息学分析结果表明，这些差异蛋白功能涉及代谢途径、核糖体、类黄酮生物合成、亚油酸代谢、氨基糖-核苷酸代谢、氨酰生物合成、亚麻酸代谢、次生代谢产物的生物合成、苯基丙酸生物合成、RNA运转、谷胱甘肽代谢、抗坏血酸盐和aldarate代谢等。功能关联分析结果表明，HR48/HR0比较组中有27条Pathway通路共关联到101个DEPs和23个DEGs，HS48/HS0比较组中有15条Pathway通路共关联到147个DEPs和16个DEGs，HR0/HS0比较组中有18条Pathway通路共关联到82个DEPs和10个DEGs，HR48/HS48比较组中仅有1条Pathway通路关联到71个DEPs和2个DEGs。同时关联发现胰蛋白酶抑制剂A、K型胰蛋白酶抑制剂、查尔酮异构酶4、脂氧合酶9、α-加氧合酶1、9S-脂氧合酶、植物凝集素前体、POD12、应激蛋白SAM22和cAPX1等可能是大豆抗豆卷叶螟潜在的靶标蛋白（基因）。qRT-PCR表达分析结果表明，5个DEPs/DEGs在HR48/HR0比较组的表达趋势与它们的RNA-Seq和iTRAQ结果基本一致。【结论】确定了一些与豆卷叶螟的抗性相关蛋白（基因）和代谢通路，这些差异蛋白可能直接或间接参与大豆对豆卷叶螟的抗虫反应。

关键词: 大豆, 豆卷叶螟, 转录组, 蛋白质组, 关联分析

Abstract: 【Objective】In order to lay a solid basis for a holistic understanding of the defense molecular mechanism of soybean resistance to Lamprosema indicata in soybean, the correlation analysis of the transcriptomic and proteomic of soybean leaves (the highly resistant line (Gantai-2-2) and the highly susceptible line (Wan 82-178)) under Lamprosema indicate larva feeding for 0 h and 48 h was conducted. 【Method】 In this study, soybeans with high resistance and susceptibility to Lamprosema indicata were selected as the research objects. The gene and protein expression abundance was analyzed for the soybean following the Lamprosema indicata feedings using RNA-Seq and iTRAQ technology, respectively. Then the correlation value between the protein level and the transcription level was calculated.【Result】There were 236 DEPs and 1 064 DEGs, 250 DEPs and 680 DEGs, 213 DEPs and 605 DEGs, 211 DEPs and 468 DEGs identified in HR48/HR0, HS48/HS0, HR0/HS0 and HR48/HS48. The results indicated that the correlations between the quantitative protein and mRNA were 0.156, 0.2687, 0.1149, and 0.035 in HR48/HR0, HS48/HS0, HR0/HS0 and HR48/HS48, and 11, 9, 3 and 4 DEPs/DEGs were identified, of which 11, 9, 2 and 3 with same expression trend, respectively. These differential expression proteins were revealed to be involved in metabolic pathways, ribosome, flavonoid biosynthesis, linoleic acid metabolism, amino sugar and nucleotide sugar metabolism, aminoacyl-RNA biosynthesis, alpha-linoleic acid metabolism, biosynthesis of secondary metabolism, phenylpropanoid biosynthesis, RNA transport, glutathione metabolism, ascorbate andaldarate metabolism. The function correlation analysis results showed that 27 pathways correlated to 101 DEPs and 23 DEGs were identified in HR48/HR0, 15 pathways correlated to 147 DEPs and 16 DEGs were identified in HS48/HS0, 18 pathways correlated to 82 DEPs and 10 DEGs were identified in HR0/HS0, and 1 pathway correlated to 71 DEPs and 2 DEGs were identified in HR48/HS48. Candidate differential expression proteins closely related to insect resistance, such as trypsin inhibitor A-like, kunitz-type trypsin inhibitor, chalcone isomerase 4-like, lipoxygenase-9, alpha-dioxygenase 1-like, linoleate 9S-lipoxygenase 1-like isoform 1, lectin precursor, peroxidase 12-like, stress-induced protein SAM22, scorbate peroxidase 1, cytosolic, and so on. Finally, qRT-PCR analysis confirmed that the two kinds of “omics” results were reliable. 【Conclusion】 In this study, some proteins and metabolic pathways were related to insect-resistant, these differential proteins were directly or indirectly involved in soybean response to Lamprosema indicata.

Key words: soybean, Lamprosema indicata, transcriptomic, proteomic, correlation analysis

曾维英，孙祖东，赖振光，蔡昭艳，陈怀珠，杨守臻，唐向民. 大豆抗豆卷叶螟的转录组和蛋白质组关联分析[J]. 中国农业科学, 2018, 51(7): 1244-1260.

ZENG WeiYing, SUN ZuDong, LAI ZhenGuang, CAI ZhaoYan, CHEN HuaiZhu, YANG ShouZhen, TANG XiangMin. Correlation Analysis on Transcriptomic and Proteome of Soybean Resistance to Bean Pyralid(Lamprosema indicata)[J]. Scientia Agricultura Sinica, 2018, 51(7): 1244-1260.

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