高级检索+

核桃品种及近缘树种鉴定的DNA条形码开发

Development of DNA Barcodes for Identification of Walnut Cultivars and Related Tree Species

  • 摘要: 为筛选适合形态学不易区分的核桃品种及近缘树种鉴定的DNA条形码,提供准确鉴定核桃品种的新途径,确保核桃产业良种化健康发展,以11个核桃品种和5个近缘种为试材,以3个叶绿体基因(rbcL、psbA-trnH、matK)和1个核DNA(ITS1)序列为候选条形码,通过对候选条形码片段的扩增、测序和序列变异分析,选择扩增效率高、品种间变异位点丰富的DNA条形码。结果表明,4条候选DNA条形码在16个供试树种(品种)中的扩增和测序成功率均为100%。rbcL序列在树种(品种)间的差异极小,psbA-trnH序列只在属间有差异。matK和ITS1序列在树种间存在差异,序列一致性分别为99.12%和99.88%,碱基位点变异数分别为18个和8个;不同树种(品种)在基于matK序列构建的系统发育树中能明显区分,验证了利用matK鉴定核桃品种及其近缘种的准确性。从而得到使用matK+ITS1条形码组合可以有效鉴别核桃品种及其近缘种。

     

    Abstract: To screen DNA barcodes suitable for the identification of walnut cultivars and related species that were not easily distinguished by morphology, so as to provide a new way for accurate identification of walnut cultivars and ensure the healthy development of walnut industry, 11 walnut cultivars and 5 relatives were used as test materials, DNA barcodes with high amplification efficiency and abundant variation sites among cultivars were selected by amplification, sequencing and sequence variation analysis in three chloroplast genes(rbcL, psbA-trnH, matK) and one nuclear DNA sequence(ITS1). The results showed that, the success rates of amplification and sequencing of the four candidate DNA barcodes in 16 species(cultivars) were 100%. The rbcL sequences showed minimal differences among species(cultivars), while the psbA-trnH sequences differed only among genera. matK and ITS1 sequences differed among species(cultivars), and their similarities were 99.12% and 99.88%, with 18 and 8 base site variants, respectively. Different species(cultivars) clustered differently in the phylogenetic tree constructed based on matK sequence, which verified the accuracy of using matK to identify walnut cultivars and their relatives. Thus, the combination of matK+ITS1 barcode can effectively identify walnut cultivars and their relatives.

     

/

返回文章
返回