新疆野兔DNA条形码筛选

The Screening of DNA Barcode for Hares in Xinjiang

旨在筛选一种用于快速鉴定新疆兔属物种的DNA条形码。本研究以新疆4种野兔共计40例样本为研究对象,以CO1、ND4、16S rRNA和ITS2为候选基因,经PCR扩增和测序后,综合分析比较候选序列在种水平上的鉴定能力。结果表明,ITS2候选片段因未获得达到测序要求的PCR扩增产物最先被排除。相较于16S rRNA,CO1和ND4在序列特征、变异特征参数和秩和检验结果上均表现出更强的变异水平,遗传频率分布图有着更宽的gap,系统聚类树的鉴定结果有着更高的鉴定准确率。而从种间变异特征、种间秩和检验结果及遗传频率分布图来看,ND4均略优于CO1。本试验综合分析得到的最优DNA条形码为ND4,候补DNA条形码为CO1。该结论可为新疆兔属物种的分类分布研究和珍稀野兔资源的保护利用等提供一定的参考依据。

The purpose of this study was to screen the optimal DNA barcode for the rapid identification of hares in Xinjiang. A total of 4 hare breeds including 40 samples were collected, CO1, ND4, 16 S rRNA and ITS2 genes were selected as candidate sequences. After PCR amplification and sequencing, the identification ability of candidate sequences at species level were analyzed and compared. The results showed that ITS2 fragments was first eliminated because no PCR amplification product was acquired for sequencing after repeated experiments. Compared with 16 S rRNA, CO1 and ND4 showed stronger variation levels in sequence characteristics, variation characteristic parameters, and rank sum test results, their genetic frequency distribution map had wider gap, and the identification results of the systematic clustering tree had higher accuracy. Further, the characteristics of interspecies variation, interspecies rank sum test results and genetic frequency distribution map all indicated that ND4 was slightly better than CO1. By comprehensive analysis, the optimal DNA barcode obtained in this study was ND4, while CO1 could serve as the alternate one. These results would provide reference for the study of distribution and classification of hares in Xinjiang and their protection and utilization.