利用DNA条形码技术鉴别22种金花茶种质资源  

Identification of 22 Kinds of Camellia nitidissima Germplasm Resources by DNA Barcoding Technology

作　　者：姚涵雅 李耀燕[1,2] 白燕远 高慧[1] 闫国跃 谢阳姣[1,2] YAO Hanya;LI Yaoyan;BAI Yanyuan;GAO Hui;YAN Guoyue;XIE Yangjiao(Guangxi University of Chinese Medicine,Nanning 530200,China;Guangxi Talent Highland for Zhuang&Yao Medicine and Combination of Medical Care and Elderly Care,Nanning 530200,China)

机构地区：[1]广西中医药大学,南宁530200 [2]广西壮瑶医药与医养结合人才小高地,南宁530200

出　　处：《种子》2023年第10期144-153,共10页Seed

基　　金：广西自然科学基金重点项目(2020GXNSFDA297029);中药学广西一流学科(桂教科研[2018]12号)。

摘　　要：收集22种金花茶新鲜叶片,运用离心柱型植物基因组试剂盒的方法提取DNA,使用ITS2、matK、rbcL、trnL-trnF、psbA-trnH序列通用引物进行PCR扩增、电泳检测后双向测序,应用核糖体DNA第二内部转录间隔区(ITS2)、RNA转录体Ⅱ型内含子剪切酶基因(matK)、叶绿体核酮糖-1,5-二磷酸羧化酶大亚基(rbcL)、叶绿体非编码区(trnL-trnF、psbA-trnH)序列对22种金花茶进行DNA条形码(DNA barcoding)分子鉴定,并确定其亲缘关系。结果表明,22种金花茶原植物样品的DNA均能成功提取,ITS2引物扩增失败,其他引物扩增成功且测序成功率均达到100%。matK、rbcL、trnL-trnF、psbA-trnH及其组合序列能将不同种金花茶成功区分。matK、rbcL、trnL-trnF、psbA-trnH及其组合序列在22种金花茶分子鉴定及确定亲缘关系方面具有较好的应用价值。22 kinds of fresh leaves of Camellia nitidissima were collected,and DNA was extracted by centrifugal column plant genome kit.ITS2,matK,rbcL,trnL-trnF,psbA-trnH sequence universal primers were used for PCR amplification,electrophoresis detection and biaxial sequencing.The sequences of ribosomal DNA second internal transcriptional spacer(ITS2),RNA transcriptor typeⅡintron cut enzyme gene(matK),chloroplast ribulose-1,5-diphosphate carboxylase large subunit(rbcL),and chloroplast non-coding region(trnL-trnF,psbA-trnH)were used to study 22 kinds of Camphor tea DNA barcoding molecular identification,and to determine the relationship.The results showed that the DNA of 22 species of Camellia nitidissima samples could be extracted successfully,the amplification of ITS2 primer failed,and the amplification of other primers was successful and the sequencing success rate reached 100%.matK,rbcL,trnL-trnF,psbA-trnH and their combination sequences can distinguish different kinds of golden tea successfully.matK,rbcL,trnL-trnF,psbA-trnH and their combination sequences had good application value in molecular identification and genetic relationship determination of 22 Camellia nitidissima species.

关 键 词：金花茶 DNA条形码 分子鉴定 亲缘关系 

分 类 号：S685.14[农业科学—观赏园艺]