引用本文:	逄锦菲,孔杰,孟宪红,栾生,罗坤,曹宝祥,刘宁.中国对虾(Fenneropenaeus chinensis) “黄海2号”人工感染WSSV的荧光定量分析.海洋与湖沼,2013,44(3):782-787.

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*中国对虾(Fenneropenaeus chinensis) “黄海2号”人工感染WSSV的荧光定量分析*
逄锦菲^1,2, 孔杰², 孟宪红², 栾生², 罗坤², 曹宝祥², 刘宁²
1.上海海洋大学水产与生命学院;2.中国水产科学研究院黄海水产研究所农业部海洋渔业可持续发展重点实验室

摘要:

采用人工投喂的方法对人工选育的104个“黄海2号”中国对虾(Fenneropenaeus chinensis)家系的5557尾个体[体重(2.1±0.7)g]进行WSSV感染实验, 分别选取存活时间最长(342h)和存活时间最短(45h)的48尾个体, 定义为抗病组和敏感组。用荧光定量PCR方法检测两组对虾体内WSSV含量, 结果表明, 抗病组和敏感组对虾体内WSSV含量范围分别为(2.64×10⁴)—(3.13×10⁶)和23.2—1.92×10⁶ copies/ng DNA, 平均值分别为8.71×10⁵和3.19×10⁵ copies/ng DNA, 抗病组对虾WSSV含量显著高于敏感组(P<0.05), 显示抗病组比敏感组具有更强的抗病性能。对感染测试的所有对虾的死亡情况进行统计分析, 结果表明, 其死亡曲线不符合数量性状的正态分布特征, 提示中国对虾对WSSV的抗性可能由少数几个主基因决定。本研究可为揭示中国对虾抗WSSV性状的基因决定机制提供依据。

关键词: 中国对虾 WSSV 人工感染荧光定量PCR

DOI：10.11693/hyhz201303037037

分类号:

基金项目:国家自然科学基金资助项目, 31072206 号, 31172402 号; 青岛市关键技术攻关类项目, 11-1-1-11-hy 号

附件

REAL-TIME PCR ASSAY FOR QUANTIFYING WSSV LOAD IN FENNEROPENAEUS CHINENSIS“HUANGHAI No.2”

PANG Jin-Fei^1,2, KONG Jie², MENG Xian-Hong², LUAN Sheng², LUO Kun², CAO Bao-Xiang², LIU Ning²

1.College of Aqua-life Science and Technology, Shanghai Ocean University;2.Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences

Abstract:

A total of 5557 shrimp individuals [(2.1±0.7)g] from 104 families of Fenneropenaeus chinensis “Huanghai No.2” obtained by selective breeding were artificially infected with WSSV. 48 samples with the longest survival time were defined as the WSSV-resistant group and 48 samples with the shortest were defined as the WSSV-susceptible group. The average survival time of the resistant and the susceptible group was 342h and 45h, with the difference in time of 297h. The WSSV loads in shrimp were quantified by real time PCR. The WSSV loads in the resistant and susceptible group were (2.64×10⁴)—(3.13×10⁶) and 23.2—1.92×10⁶ copies/ng DNA, with the average of 8.71×10⁵ and 3.19×10⁵ copies/ng DNA, respectively. WSSV loads of the resistant group was significantly higher than the susceptible group (P<0.05), which showed higher disease resistance ability of resistant group. The death curve of the shrimp didn’t accord with the normal distribution of quantitative trait, suggesting that the trait of WSSV resistance may be controlled by several major genes. This study will be helpful to reveal the genetically determined mechanism of WSSV-resistance trait of F. chinensis.

Key words: Fenneropenaeus chinensis WSSV artificial infection, Real-time PCR