“黄海1号”中国明对虾抗逆性状SRAP标记  被引量:3

Sequence-related amplified polymorphism markers related to stress resistance traits in "Huanghai No.1" Fenneropenaeus chinensis

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作  者:陈华增[1,2] 李健[1] 王清印[1] 何玉英[1] 李吉涛[1] 戴芳钰[1] 王学忠 

机构地区:[1]中国水产科学研究院黄海水产研究所,山东青岛266071 [2]上海海洋大学水产与生命学院,上海201306 [3]山东省昌邑市海丰水产养殖有限责任公司,山东潍坊262600

出  处:《中国水产科学》2011年第6期1243-1249,共7页Journal of Fishery Sciences of China

基  金:国家自然科学基金资助项目(40706052);国家科技支撑计划专题(2006BAD01A13);农业部公益性行业专项(nyhyzx07-042);国家虾产业技术体系资助项目(nycytx-46)

摘  要:采用序列相关扩增多态性(sequence-related amplified polymorphism,SRAP)分子标记技术,结合分群分析法(bulk segregate analysis,BSA)对中国明对虾(Fenneropenaeus chinensis)高氨氮和高pH胁迫敏感组和耐受组进行分析研究,筛选出与高氨氮或高pH耐受性相关的遗传标记。应用110对SRAP引物进行筛选研究,根据在BSA基因池产生的差异,筛选出与高氨氮耐受相关引物77对,与高pH耐受相关引物102对,以用于验证分析。根据片段在群体中出现频率和变化规律,筛选出6个可能与高氨氮耐受性相关的分子遗传标记,其中耐受高氨氮负相关标记1个,正相关标记5个;7个可能与高pH耐受性相关的分子遗传标记,其中耐受高pH负相关标记2个,正相关标记5个。对获得的13个序列片段进行回收,连接于pMD-18T载体后转化于大肠杆菌TOP10感受态细胞,进行了克隆和测序。将测序结果进行了BLAST分析比对,发现测得片段序列与数据库中序列同源性较低(一般都低于15%),未找到与之同源性较高的功能基因,推论这些特异性序列片段标记可能与高氨氮或高pH耐受性性状密切相关,后续群体验证工作正在进行,以期筛选出与中国明对虾抗逆性状密切相关的序列特征性片段扩增区域(sequence characterized amplified region,SCAR)分子标记,为分子标记辅助育种提供技术支持。The sequence-related amplified polymorphism (SRAP) molecular marker technique and BSA were used to investigate molecular markers related to stress resistance traits in Fenneropenaeus chinensis. The markers, which were based on differences between sensitive and tolerant samples, will be used to provide information and tools for the protection of the F. chinensis germplasm resource and for molecular marker-assisted breeding. We used 110 SRAP primers to screen for molecular markers. We selected 77 and 102 SRAP primers to screen ammonia nitrogen and pH-BSA gene pools, respectively. According to the frequency and pattern of bands in the samples, six markers associated with resistance to high ammonia nitrogen were selected; one from sensitive samples and five from tolerant samples. These markers ranged in length from 166 to 571 bp. Similarly, seven markers associated with resistance to high pH were selected; two from sensitive samples and five from tolerant samples. These 13 genetic markers were cloned into the pMD-18T vector, transformed into Escherichia coli TOP10, and sequenced. The sequence data has been submitted to GenBank under the accession numbers GU570681–GU570693. Sequence and BLAST analyses showed that there was low similarity (〈15%) between the markers and known functional genes, indicating that these markers are tightly linked to stress resistant traits but are not functional genes. Further research has been carried out to validate these results. These markers can be used to screen for stress resistance traits in F. chinensis, and provide a valuable tool for marker-assisted selection breeding.

关 键 词:中国明对虾 氨氮 PH SRAP 分群分析法 抗逆性状 

分 类 号:S96[农业科学—水产养殖]

 

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