亚洲玉米螟中stuxnet基因的时空表达与功能分析
Spatiotemporal expression and functional analysis of the Ostrinia furnacalis stuxnet gene
吴小宇 杨兴卓 梁 佳 赵贤国 赵章武 杜 娟
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DOI：10.7679/j.issn.2095-1353.2021.091
作者单位：中国农业大学植物保护学院，农业部作物有害生物监测与绿色防控重点实验室，北京 100193；中国农业大学植物保护学院，农业部作物有害生物监测与绿色防控重点实验室，北京 100193；中国农业大学植物保护学院，农业部作物有害生物监测与绿色防控重点实验室，北京 100193
中文关键词：亚洲玉米螟；stuxnet；多梳蛋白；时空表达；功能分析
英文关键词：Ostrinia furnacalis; stuxnet; polycomb-group; spatiotemporal expression; functional analysis
中文摘要：
【目的】 本研究旨在鉴定亚洲玉米螟Ostrinia furnacalis中stuxnet (stx)同源基因Ofstuxnet (Ofstx)，进一步明确Ofstx基因的功能，探索基于表观遗传调控的害虫防治新策略。【方法】 基于反转录技术获取亚洲玉米螟Ofstx基因的全长cDNA序列，进行cDNA全长测序，分析其生物学信息；分别提取亚洲玉米螟不同发育阶段（3-5龄期幼虫）及不同组织（头、中肠、脂肪体）中RNA，并通过RT-qPCR技术检测Ofstx基因在亚洲玉米螟不同发育阶段及不同组织中的相对表达量。【结果】 亚洲玉米螟Ofstx基因全长2 112 bp，编码蛋白质含703个氨基酸，预测分子量76.98 ku，等电点8.30。比对其它物种Stx同源蛋白氨基酸序列发现其都含有UBL保守结构域。系统发育树分析发现与草地贪夜蛾Spodoptera frugiperda所属的夜蛾科同源性较高。qPCR结果显示Ofstx在脑内相对表达量最高，在3-5龄幼虫中，4龄幼虫体内总体表达量最高。【结论】 Ofstx基因主要在亚洲玉米螟脑内表达，幼虫阶段的第4龄期相较第3、第5龄期相对表达高，可能参与调控昆虫生长发育，具体功能有待进一步研究。
英文摘要：
[Objectives] The aim
of this study was to identify the Asian corn borer Ostrinia furnacalis stuxnet (stx) gene, Ofstuxnet (Ofstx), clarify its
function and investigate the potential of using this gene to develop a novel
pest control strategy based on epigenetic regulation. [Methods] The full-length cDNA sequence of the Ofstx gene
was obtained using the reverse transcription technique, and its gene sequence
obtained by full-length cDNA sequencing. RNA was extracted from 3rd to 5th instar larvae and from different tissues (head, midgut, fat
body), and the relative expression of the Ofstx gene in these age groups
and tissues was detected using RT-qPCR. [Results] The total length of the Ofstx gene was
2 112 bp, The encoding protein contained 703 amino acids, has a predicted
molecular weight of 76.98 ku, and an isoelectric point of 8.30. Amino acid
sequence comparison of the Ofstx gene and homologous
proteins from other species showed that they all contained an UBL conserved
domain. A phylogenetic tree shows that the Ofstx gene is highly
homologous with Noctuid stx genes，including that of Spodoptera frugiperda. qPCR
results show that relative expression of Ofstx was highest in the head
and that overall expression was highest in 4th instar larvae. [Conclusion] The stx gene is mainly expressed
in the head of O. furnacalis, and in fourth instar larvae than in third and
fifth instar larvae. This suggests that this gene may be involved in the
regulation of growth and development. Further research is required to reveal
its specific functions.