短管赤眼蜂Trichogramma pretiosum microRNA鉴定及四环素响应microRNA功能分析
Identification of microRNAs in Trichogramma pretiosum and functional analysis of tetracycline-responsive microRNAs
肖安琪1, 2** 沈家安1 李迎瑞1 吴 萌1 曹 睿3 董 旭1 张海燕1, 2, 3*** 刘聪鹤1, 2, 3*** 李 鹏4
点击：90次 下载：4次
DOI：10.7679/j.issn.2095-1353.2026.080
作者单位：1. 黑龙江八一农垦大学农学院，大庆 163319；2. 农业农村部东北平原农业绿色低碳重点实验室，大庆 163319；3. 黑龙江省秸秆资源化利用工程技术研究中心，大庆 163319；4. 黑龙江省植检植保站，哈尔滨 150090
中文关键词：短管赤眼蜂；microRNA；全转录组；孤雌产雌；生殖机制
英文关键词：Trichogramma pretiosum; microRNA; whole transcriptome; parthenogenesis; reproductive mechanism
中文摘要：【目的】 赤眼蜂是农林害虫的重要卵寄生蜂，感染沃尔巴克氏菌Wolbachia后其生殖方式由两性生殖（Bisexual reproduction）转为孤雌产雌生殖（Thelytoky），四环素（Tetracycline）可以去除赤眼蜂体内的共生菌使孤雌产雌的赤眼蜂恢复两性生殖方式。本研究旨在探究与短管赤眼蜂Trichogramma pretiosum生殖相关的功能性miRNA及其靶基因。【方法】 以2.5 mg/mL盐酸四环素蜂蜜溶液连续饲喂处理短管赤眼蜂孤雌产雌品系3代成蜂，计算寄生率、羽化率和雌性比，利用Illumina平台进行短管赤眼蜂孤雌产雌品系成蜂体内small RNA转录组测序，筛选差异表达基因并进行GO/KEGG/COG分析；利用RT-qPCR检测短管赤眼蜂转录组数据中差异表达的10个基因（novel_miR_124、novel_miR_311、novel_miR_ 176、novel_miR_ 99、novel_miR_510、novel_miR_48、novel_miR_96、novel_miR_315、novel_miR_280、novel_miR_92）的表达量。【结果】 本研究测定了四环素处理前后短管赤眼蜂的全转录组，获得短管赤眼蜂差异表达microRNA 79个，上调表达67个，下调表达12个，共预测靶基因4 129个，其中表达量有差异的基因为1 224个。GO分析表明，靶基因参与了转录调控（Regulation of transcription，DNA-templated）、跨膜运输（Transmembrane transport）、催化活性（Catalytic activity）等功能。KEGG分析表明，ECM、神经活性配体-受体相互作用、Mapk以及TGF-beta和Hippo等信号通路可能在短管赤眼蜂感染沃尔巴克氏菌后的孤雌生殖机制调节中发挥关键作用。【结论】 本研究表明，novel_miR_280、novel_miR_99、novel_miR_48和novel_ miR_311靶基因及信号通路是调控短管赤眼蜂孤雌产雌的关键，并且减数分裂和第一次有丝分裂的异常对其生殖机制的转变起着不可或缺的作用。此外，本研究还为该物种生殖机制改变后的表观遗传研究提供了全面的转录组参考信息。
英文摘要：[Aim] To explore functional miRNAs and their target genes related to the reproduction of Trichogramma pretiosum, an important egg parasitoid of agricultural and forestry pests. [Methods] The third generation of T. pretiosum parthenogenetic strain was continuously fed with 2.5 mg/mL tetracycline hydrochloride honey solution. Parasitic rate, eclosion rate and female ratio were calculated.The Illumina platform was used to sequence the small RNA transcriptome of T. pretiosum parthenogenetic strain adults. Differentially expressed genes were screened and GO/KEGG/COG analysis was performed. The expression levels of 10 differentially expressed genes ( novel_miR_124, novel_miR_311, novel_miR_176, novel_miR_99, novel_miR_510, novel_miR_48, novel_miR_96, novel_miR_315, novel_miR_280, novel_ miR_92) in the transcriptome data of T. pretiosum were detected by RT-qPCR. [Results] The entire transcriptome of T. pretiosum was sequenced and a total of 79 differentially expressed microRNAs identified, 67 of which were up-regulated and 12 down-regulated. A total of 4 129 target genes were predicted, of which, 1 224 were differentially expressed. GO analysis indicated that these target genes were involved in transcriptional regulation, transmembrane transport, catalytic activity, and other functions. KEGG analysis suggests that the ECM, neuroactive ligand-receptor interaction, Mapk, and TGF-beta and Hippo, signaling pathways play key roles in regulating thelytoky in T. pretiosum after Wolbachia infection. [Conclusion] This study demonstrates that the target genes and signaling pathways of novel_miR_280, novel_miR_99, novel_miR_48, and novel_miR_311, are crucial for regulating thelytoky in T. pretiosum, and that abnormal meiosis and first mitosis are indispensable for the transformation of its reproductive mechanism. Additionally, this study provides a comprehensive transcriptome reference information for epigenetic research on the reproductive mechanism of this species after alteration.