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基因条码在条斑紫菜(Pyropia yezoensis)不同品系间种质鉴定中的适用性
曹殿嘉1, 严兴洪1,2
1.上海海洋大学水产与生命学院 上海 201306;2.上海海洋大学省部共建水产种质资源发掘与利用教育部重点实验室 上海 201306
摘要:
为寻找适宜的条斑紫菜(Pyropia yezoensis)品系间种质分子标记鉴定法,利用4种基因条码(RUBISCO Spacer、UPA、Cox2-3和ITS-5.8S)对产自中国和日本的6个条斑紫菜品系的亲缘关系进行了分析研究。结果显示,在6个条斑紫菜品系中,上述4种基因条码的序列同源性较高,同源性为97.8%–100%,其中,RUBISCO Spacer序列的同源性为100%。UPA序列的同源性分析结果显示,在中国产的LS、B4、HT和TM18品系中,该序列完全相同,但与日本产的GT和ROS-2两个品系的序列有差异,该序列的同源性为99.2%。在6个品系中,Cox2-3和ITS-5.8S序列的同源性相对较低,分别为97.8%–99.2%和98.4%–99.8%。根据UPA、Cox2-3和ITS-5.8S基因条码通过邻接法构建的系统进化树显示,依据ITS-5.8S和UPA序列均能分别区分中国产的4个品系(LS、B4、HT和TM18)与日本产的两个品系(GT和ROS-2),分别形成两个大分支,其聚类结果与实际亲缘关系一致。但利用ITS-5.8S序列均能对中国产的4个品系和日本产的两个品系进行再区分,而利用UPA序列却无法对中国产的4个品系进行再区分,只能区分日本产的两个品系。使用Cox2-3序列则将GT、ROS-2、LS和TM18这4个品系聚成一个大分支,其聚类结果与实际亲缘关系不一致。上述结果说明,利用ITS-5.8S序列对本研究的6个条斑紫菜品系进行种质鉴定,效果最好。
关键词:  条斑紫菜  基因条码  品系  种质鉴定
DOI:10.11758/yykxjz.20150430002
分类号:
基金项目:国家高科技研究发展计划(863计划)项目(2012AA10A411)、国家自然科学基金项目(31072208)、国家海洋局公益专项(201105008;201105023)、国家农业科技成果转化资金项目(2013GB2C220537)、农业部公益性专项(200903030)、上海市科委重点科技攻关项目(10391901100)、上海高校水产学一流学科建设项目、福建省科技重大专项(2011NZ0001-1)和福建省省长专项基金(2014S1477-10)共同资助
Applicability of DNA Barcoding in the Identification of Different Strains in Pyropia yezoensis
CAO Dianjia1, YAN Xinghong1,2
1.College of Fisheries and Life Sciences, Shanghai Ocean University, Shanghai 201306;2.Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Shanghai Ocean University, Ministry of Education, Shanghai 201306
Abstract:
In order to find out an effective method to identify Pyropia yezoensis germplasm, we tested the efficacy of four common DNA barcodings—RUBISCO Spacer, UPA, Cox2-3, and ITS-5.8S—in determining the genetic relationships between six strains of P. yezoensis from China and Japan. The four DNA barcodings showed high sequence homologies varying from 97.8% to 100%, and the homology of RUBISCO Spacer was 100%. Four Chinese strains (LS, B4, HT, and TM18) shared the same UPA sequence. However, the barcoding was different from that of the Japanese strains GT and ROS-2 and the homology was 99.2%. The homologies of Cox2-3 and ITS-5.8S sequences were 97.8%-99.2% and 98.4%-99.8% respectively, which were lower than those of RUBISCO Spacer and UPA. Phylogenesis of UPA, Cox2-3 and ITS-5.8S sequences revealed that the four Chinese strains clustered into one branch according to their UPA and ITS-5.8S sequences, whereas the two Japanese strains belonged to another branch. These results were consistent with their actual relationships. Moreover, the four Chinese strains could be further distinguished according to their ITS-5.8S sequences, but were identified as in the same branch with the Cox2-3 barcoding. The results confirmed that ITS-5.8S was the most effective barcoding for identifying P. yezoensis strains in this study
Key words:  Pyropia yezoensis  DNA barcodings  Strains  Germplasm identification