基于DNA条形码技术构建王朗国家级自然保护区陆生脊椎动物遗传资源数据库及物种鉴定

邢超, 林依, 周智强, 赵联军, 蒋仕伟, 林蓁蓁, 徐基良, 詹祥江

The establishment of terrestrial vertebrate genetic resource bank and species identification based on DNA barcoding in Wanglang National Nature Reserve

Chao Xing, Yi Lin, Zhiqiang Zhou, Lianjun Zhao, Shiwei Jiang, Zhenzhen Lin, Jiliang Xu, Xiangjiang Zhan

表1 用于PCR扩增和DNA测序的引物

Table 1 Primers used for PCR amplification and DNA sequencing

类别 Category 目标片段所在基因(引物名称) Target gene (primers' names) 上游引物序列5'-3' Forward primer sequence 5'-3' 下游引物序列5'-3' Reverse primer sequence 5'-3' 退火温度 Annealing temperature (℃) 循环 次数 Cycles 参考文献 Reference 兽类 Mammals COI (COIung) GTACCGCTAATAATTGGTGCTCC GGGTGGCCAAAGAATCAGAACAAGTG 56 35 Kumar et al, 2017 16S (16SF/R) GAGAAGACCCTATGGAGC ATAGAAACCGACCTGGAT 50 30 Xiong et al, 2016 COI (BatL/R6036R) CCTACTCRGCCATTTTACCTATG ATCTCTGGGTGTCCAAAGAATCA 55 35 Robins et al, 2007 ATP6 (ATP6-DF3/ATP6-DR2) AACGAAAATCTATTCGCCTCT TGGATGGACAGTATTTGTTTTGAT 60/50 10/30 Chaves et al, 2012 D-loop (BEDL225/H470) ATGTACATACTGTGCTTGGC GTCATTAGTCCATCGAGATG 53 38 Zhang et al, 2002 鸟类 Avians COI (COI-K_Bird_F/R) CCCCAGACATAGCATTYCC TTGTGATAGTGGTGGGGTTTTAT 46/53 5/45 Parejo-Farnés et al, 2018 Cytb (Cytb-L14816/H15173) CCATCCAACATCTCAGCATGATGAAA CCCCTCAGAATGATATTTGTCCTCA 50 30 Parson et al, 2000 COI (L6697Bird_F/H7390Thrush_R) TCAACYAACCACAAAGAYATCGGYAC ACGTGGGARATRATTCCAAATCCTG 48/51 5/30 Saitoh et al, 2015 COI (PasserF/R) CCAACCACAAAGACATCGGAACC GTAAACTTCTGGGTGACCAAAGAATC 58 35 Lohman et al, 2009 两栖类 Amphibians COI (LCO1490/HCO2198) GGTCAACAAATCATAAAGATATTGG TAAACTTCAGGGTGACCATCA 50 35 Folmer et al, 1994 COI (LepF/R) ATTCAACCAATCATAAAGATATTGG TAAACTTCTGGATGTCCATCA 46 35 Hebert et al, 2004a 16S (P7/8) CGCCTGTTTACCCAT CCGGTCTGAACTCAGATCACGT 55 36 Simon et al, 1994 16S (Sar/Sbr) CGCCTGTTTATCCAT CCGGTCTGAACTCAGATCACGT 48 40 Palumbi et al, 2002 12S-16S (1602L/2571H) GTATACCGGAAGGTGTACTTGGAACAG TACCTTCGCACGGTCAGAATACCGC 50 35 Fu et al, 2007 Cytb (FrogGlu-f/Thr-r) TGATCTGCCACCGTTG CTCCATTCTTCGRCTTACAAG 46 40 Hillis et al, 1996 RAG1 (RAG1-F/R) AGCTGCAGYCARTACCAYAARATGTA GCAAAGTTTCCGTTCATTCTCAT 50 35 Fu et al, 2007