实时荧光定量PCR检测凡纳滨对虾和罗氏沼虾卵黄蛋白原mRNA在卵巢和肝胰腺中的表达

Quantitative analysis of vitellogenin mRNA expression in Litopenaeus vannamei and Macrobrachium rosenbergii

  • 摘要: 本研究应用SYBR GreenⅠ荧光定量PCR法检测凡纳滨对虾和罗氏沼虾卵巢不同发育时期卵巢和肝胰腺两种组织中卵黄蛋白原mRNA的表达水平。根据卵巢发育情况把凡纳滨对虾的卵巢分为:卵原细胞增殖期,卵黄发生前期,初级卵黄发生期,次级卵黄发生期,成熟期和恢复期。罗氏沼虾的卵巢分为:卵黄发生前期,初级卵黄发生期,次级卵黄发生期,成熟期和抱卵期。研究结果表明:凡纳滨对虾和罗氏沼虾的卵巢和肝胰腺中都有卵黄蛋白原mRNA的表达。随着卵巢的发育,凡纳滨对虾卵巢中卵黄蛋白原mRNA的相对表达量在前5个阶段不断增加,分别为:1.1, 5.9, 10.4, 26.9, 85.0,恢复期急剧减少,为1.6。肝胰腺中的相对表达量也不断增加,分别为:1.3, 3.3, 7.1, 37.3, 51.6,恢复期急剧减少,为1.0。罗氏沼虾肝胰腺中卵黄蛋白原mRNA的相对表达量在前四个阶段不断增加,分别为:3.4, 12.6, 15.2,38.9,抱卵期急剧减少,为2.9,而卵巢在整个发育过程中对卵黄蛋白原合成的贡献比较小,分别为1.0, 1.3, 1.7, 4.8, 1.5。

     

    Abstract: The accurate site for vitellogenin synthesis was investigated in two representative shrimp species Litopenaeus vannamei and Macrobrachium rosenbergii with real-time PCR in this experiment. Based on histological observation, the ovarian development was divided into six stages in L. vannamei (oogonium proliferation, previtellogenic, primary vitellogenic, secondary vitellogenic, mature, and recovery stages) and five stages in M. rosenbergii (previtellogenic, primary vitellogenic, secondary vitellogenic, mature, and gravid stages). It was found that both L. vannamei and M. rosenbergii expressed Vg-mRNA in hepatopancreas and ovary. In L. vannamei, the relative quantity of Vg-mRNA expression of each developmental stage was 1.1, 5.9, 10.4, 26.9, 85.3, and 1.5 in ovary, and 1.3, 3.2, 7.1, 37.2, 51.6, and 1.0 respectively in hepatopancreas. In M. rosenbergii, the relative quantity of Vg-mRNA expression of hepatopancreas in each ovarian developmental stage was 3.4, 12.6, 15.2, 38.9, and 2.9 compared to ovary 1.0, 1.3, 1.7, 4.8, and 1.5, respectively. In all examined shrimps the Vg-mRNA expression decreased sharply to the minimum at the last development stage.

     

/

返回文章
返回