| 摘要: |
| 【目的】对花绒寄甲线粒体合成调控基因进行表达分析，为深入揭示花绒寄甲生殖生理活动奠定一定的理论基础。【方法】从花绒寄甲转录组文库和线粒体基因组中筛选出PGC1-α、NRF1、mtTFA、COXⅠ和ATP6基因，对其开放阅读框不完整序列进行5′RACE克隆扩增，对拥有完整开放阅读框的序列直接进行PCR扩增。用实时荧光定量qPCR分析5个线粒体合成基因在花绒寄甲成虫不同组织、不同虫态和不同虫龄中的表达差异。【结果】获得核基因编码序列3条（PGC1-α、NRF1和mtTFA）和线粒体基因编码序列2条（ATP6和COXⅠ）。花绒寄甲各基因的氨基酸序列与其他物种相应氨基酸序列同源性较高，其中mtTFA和NRF1与鞘翅目昆虫赤拟谷盗（Tribolium castaneum）相应序列的同源性高达80%以上。花绒寄甲5个线粒体合成相关基因的表达存在显著的组织特异性和虫态特异性，其在花绒寄甲精巢和脂肪体中的表达量明显高于其他组织；同时，各基因在1龄幼虫中的表达量高于其他幼虫期，在蛹中的表达量最小。花绒寄甲成虫羽化20个月后，各线粒体合成调控基因的表达量随着虫龄的增加呈上升趋势，在30个月达到最大值。【结论】线粒体合成相关基因普遍存在于花绒寄甲不同组织、虫态及虫龄中，但表达量有较大差异。核基因编码的PGC1-α、NRF1和mtTFA表达量高的组织、虫态和虫龄中，相应的线粒体编码基因COXⅠ和ATP6的表达量也相对较高。 |
| 关键词: 花绒寄甲 线粒体基因组 细胞核基因组 定量表达 线粒体合成 |
| DOI： |
| 分类号: |
| 基金项目:国家自然科学基金项目(31170608) |
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| Sequence and spatio-temporal expression of 5 mitochondrial biogenesis related genes in Dastarcus helophoroides |
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ZHANG Zhengqing,LI Menglou
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| Abstract: |
| 【Objective】The expressions of mitochondrial synthesis related genes were analyzed to provide basis for deeply understanding reproductive physiology activity of Dastarcus helophoroides.【Method】PGC1-α,NRF1,mtTFA,COXⅠ and ATP6 genes were selected from D.helophoroides transcriptome and mitochondrial genome,the 5′RACE of sequences with uncompleted open reading frames were cloned and amplified,and the sequences with completed open reading frames were amplified with PCR.The expressions of the five mitochondrial biogenesis related genes in different tissues of adult,different development stages and at different adult ages after emergence were studied by real-time fluorescence quantification qPCR.【Result】Three nDNA coding sequences (PGC1-α,NRF1and mtTFA) and two mtDNA coding sequences (ATP6 and COXⅠ) were obtained.Different amino acid sequences of D.helophoroides genes were clustered in same branch with the corresponding sequences of other species.D.helophoroides mtTFA and NRF1 shared high homology (80%) with Tribolium castaneum.The expressions of five mitochondrial biogenesis related genes existed significant tissue specificity and development stage specificity.The expression levels of these genes in testis and fat body were higher than that in other tissues.The expressions of five mitochondrial biogenesis related genes in 1 instar larva were higher rather than in other instars,with the lowest level in pupa.Twenty months after D.helophoroides adult emergence,expressions of these genes were increased with the extension of emergence,and reached the maximum after 30 months.【Conclusion】Mitochondrial synthesis related genes existed universally in D.helophoroides tissues,stages and adult ages,with great differences.The tissues,stages and adult ages with high nDNA coding genes (PGC1-α,NRF1 and mtTFA) also had high expressions of mtDNA coding genes (COXⅠ and ATP6). |
| Key words: Dastarcus helophoroides mitochondrial genome nuclear genome quantitative expression mitochondrial synthesis |
