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| 东方蜜蜂微孢子虫lncRNA5334及其顺式调控基因的表达模式 |
| Expression patterns of lncRNA5334 and its cis-acting genes in Nosema ceranae |
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| DOI： |
| 中文关键词: 东方蜜蜂微孢子虫 长链非编码RNA 顺式作用 意大利蜜蜂 上下游基因 |
| 英文关键词:Nosema ceranae long-stranded noncoding RNA cis-acting Apis mellifera ligustica upstream and downstream genes |
| 基金项目: |
| Author Name | Affiliation | | KANG Jing1#, DONG Shu-Nan1#, LI Zheng-Yuan1, WANG Yong-Jie1, MI Shi-Yu1, QIU Jian-Feng1,2,3, ZHANG Rong-Hua2,4, CHEN Da-Fu1,2,3, YAN Ti-Zhen2,4, LUO Qing-Ming2,4*, GUO Rui1,2,3* | |
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| 中文摘要: |
| 【目的】本研究旨在揭示东方蜜蜂微孢子虫Nosema ceranae侵染意大利蜜蜂Apis mellifera ligustica工蜂过程中lncRNA5334的表达模式和调控作用，为进一步探究lncRNA5334的作用机制提供基础。【方法】通过RT-PCR验证东方蜜蜂微孢子虫lncRNA5334的表达。利用相关软件预测lncRNA5334顺式调控的靶基因。采用RT-qPCR检测lncRNA5334及其靶基因在侵染过程中的相对表达量。【结果】结果显示，在接种东方蜜蜂微孢子虫的意大利蜜蜂工蜂1dpi(1 day post inoculation)、2 dpi，3dpi和4 dpi的中肠样品及东方蜜蜂微孢子虫孢子样品中均扩增出符合预期大小（约185 bp）的目的片段，而在未接种孢子的工蜂中肠中未检测到目的片段；相较于1 dpi，lncRNA5334的表达量在2 dpi下调（P>0.05），在3 dpi和 4 dpi均显著下调（P<0.05），总体呈持续下降的表达趋势；lncRNA5334潜在调控4个上下游基因（Pol II，Trx，ACS和GPDH）的转录；相较于1 dpi，lncRNA5334的表达水平在2 dpi下调（P>0.05），在3 dpi和4 dpi均显著下调（P<0.05）；Pol II的表达量在2 dpi和3 dpi上调（P>0.05），在4 dpi显著上调（P<0.05）；Trx的表达量在2 dpi，3 dpi和4 dpi皆显著上调（P<0.05）；ACS的表达量在2 dpi和4 dpi均上调（P>0.05），在3 dpi显著上调（P<0.05）；GPDH的表达量在2 dpi下调（P>0.05），在3 dpi和4 dpi均显著上调（P<0.05）。【结论】研究结果表明lncRNA5334在东方蜜蜂微孢子虫孢子及侵染过程中均表达，lncRNA5334可能通过顺式作用调控Pol II，Trx，ACS和GPDH的转录进而调节东方蜜蜂微孢子虫侵染意大利蜜蜂工蜂的过程。 |
| 英文摘要: |
| 【Aim】The aim of this study is to reveal the expression pattern and regulatory role of lncRNA5334 during the process of Nosema ceranae infection of Apis mellifera ligustica worker, providing a basis for further investigation of the regulatory mechanism of lncRNA5334. 【Methods】The expression of lncRNA5334 in Nosema ceranae was verified by RT-PCR. Relevant software were utilized to predict the target genes regulated by lncRNA5334 in cis-acting manner. RT-qPCR was used to detect the relative expression of lncRNA5334 and its target genes during the infestation process. 【Results】The results showed that the target fragment of the expected size (approximately 185 bp) was amplified from the midgut samples of Apis mellifera ligustica workers at 1 dpi (1 day post inoculation), 2 dpi, 3 dpi, and 4 dpi with Nosema ceranae, while no target fragment was detected in the midgut samples of uninoculated workers; as compared with that at 1 dpi, the expression level of lncRNA5334 was downregulated at 2 dpi (P>0.05) and significantly downregulated at 3 dpi and 4 dpi (P<0.05), presenting an overall trend of continuous decrease in expression; lncRNA5334 potentially regulated the transcription of four up- and down-stream genes (Pol II, Trx, ACS, and GPDH); as compared with that at 1 dpi, the expression level of lncRNA5334 was down-regulated at 2 dpi (P>0.05) and significantly down-regulated at both 3 dpi and 4 dpi (P<0.05), the expression level of Pol II was up-regulated at 2 dpi and 3 dpi (P>0.05) and significantly up-regulated at 4 dpi (P<0.05), the expression level of Trx was significantly up-regulated at 2 dpi, 3 dpi and 4 dpi (P<0.05), the expression level of ACS was up-regulated at 2 dpi and 4 dpi (P>0.05) and significantly up-regulated at 3 dpi (P<0.05), the expression level of GPDH was down-regulated at 2 dpi (P>0.05) while significantly up-regulated at both 3 dpi and 4 dpi (P<0.05). 【Conclusion】These results suggest that lncRNA5334 is expressed in the spore and during the infection process of Nosema ceranae, and lncRNA5334 may potentially modulate the process of Nosema ceranae infection in Apis mellifera ligustica through cis-acting manner through modulating the transcription of Pol II, Trx, ACS, and GPDH. |
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