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脊尾白虾不同发育期mtDNA拷贝数变化特征分析
张培,徐莞媛,李志辉,高焕,张庆起,赖晓芳,陈建华,阎斌伦
1.江苏省海洋生物技术重点实验室 淮海工学院海洋生命与水产学院 连云港 222005;2.江苏省海洋生物产业技术协同创新中心 连云港 222001;3.江苏省海洋资源开发研究院 连云港 222005;4.江苏省农业种质资源保护与利用平台 南京 210014;5.连云港赣榆佳信水产开发有限公司 连云港 222100
摘要:
为了解脊尾白虾(Exopalaemon carinicauda)在不同发育期线粒体基因组(mtDNA)拷贝数的变化特征,本研究共采集其8个时期(受精卵期、溞状幼体Ⅰ期、溞状幼体Ⅱ期、溞状幼体Ⅲ期、溞状幼体Ⅳ期、溞状幼体Ⅴ期、仔虾期和成虾期)的DNA样品,利用TaqMan实时荧光定量PCR技术,对上述各发育期单个细胞中的mtDNA拷贝数进行了测定。检测结果显示,脊尾白虾受精卵期、溞状幼体Ⅰ期、溞状幼体Ⅱ期、溞状幼体Ⅲ期、溞状幼体Ⅳ期、溞状幼体Ⅴ期、仔虾期和成虾期的mtDNA拷贝数的均值分别为2366、2648、2644、2873、3559、9948、6452和8872。进一步统计分析结果显示,mtDNA拷贝数与发育时间存在正相关性,相关系数为0.83。研究表明,随着脊尾白虾不断生长,其单个细胞中mtDNA拷贝数总体呈上升趋势。
关键词:  脊尾白虾  TaqMan探针法  线粒体基因组  不同发育期  拷贝数
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Profiles of mtDNA Copy Number at Different Developmental Stages of Exopalaemon carinicauda
ZHANG Pei1,2,3,4, XU Wanyuan1,2,3,4, LI Zhihui1,2,5, GAO Huan1,2,5, ZHANG Qingqi6, LAI Xiaofang1,2,3,4, CHEN Jianhua1,2,3,4, YAN Binlun1,2,3,4
1.Jiangsu Key Laboratory of Marine Biotechnology, College of Marine Life and Fisheries, Huaihai Institute of Technology, Lianyungang 222005;2.Co-Innovation Center of Jiangsu Marine Bio-Industry Technology, Lianyungang 222001;3.Marine Resource Development Institute of Jiangsu, Lianyungang 222005;4.Jiangsu Provincial Platform for Conservation and Utilization of Agricultural Germplasm, Nanjing 210014;5.Marine Resource Development Iƒƒƒ „„田ᐰ￿￿넵†‡ ¦ĀĀ‰᳀ᏃŠ넵‹ ⯨ቸ;6.Ganyu-Jiaxin Aquaculture Limited Company of Lianyungang, Lianyungang 222100
Abstract:
Mitochondria are important organelles in cells. Mitochondria can produce ATP and provide energy for almost all biological activities. Mitochondria have a relatively independent genetic system, and nuclear genes also encode certain enzymes responsible for mitochondrial DNA (mtDNA) replication and transcription. Therefore, the number and function of mitochondria is essential for embryonic development. Mitochondrial function depends on the integrity of the mitochondrial genome or the regulation of nuclear genome, as well as the mtDNA copy number. Many studies have investigated mtDNA copy numbers in animals, including studies on oocyte maturation, mammalian embryonic development, and human disease, but none have reported the mtDNA copy number in Exopalaemon carinicauda. To study the profiles of mtDNA copy number at different developmental stages of E. carinicauda, and the energy expenditure of tissues at the different stages of development, total DNA from zygote, zoea Ⅰ, zoea Ⅱ, zoea Ⅲ, zoea Ⅳ, zoea Ⅴ, post larvae, and adult shrimps were used to analyze the mtDNA copy number. One TaqMan probe-based real-time PCR was established to assess the mtDNA copy number. Our results showed that the average copy number of mtDNA per cell was 2366, 2648, 2644, 2873, 3559, 9948, 6452, and 8872 in the zygote, zoea Ⅰ, zoea Ⅱ, zoea Ⅲ, zoea Ⅳ, zoea Ⅴ, post larvae, and adults, respectively. These results show that mtDNA copy number is positively correlated with age, and the correlation coefficient (R) was 0.83, i.e. the copy numbers of mtDNA tend to increase with progressing developmental stages of E. carinicauda.
Key words:  Exopalaemon carinicauda  TaqMan real-time PCR  Mitochondrial genome  Different developmental stages  Copy number