Sequence Analysis of Different Flower Color of Bletilla striata Based on Four DNA Barcoding Markers

doi:10.14088/j.cnki.issn0439-8114.2018.17.027

Abstract

Abstract: Genomic DNA was extracted from the two kinds of color of Bletilla striata by using the CTAB method. To analysis the sequence of different flower color of Bletilla striata based on the four kinds of DNA barcode for further identification. Four DNA barcoding markers（trnH-psbA,rbcL,ITS and LSU D1-D3） were selected for PCR. PCR product was ligated into T vector for sequencing. The sequencing results were analyzed by using DNAMAN software. According to the SNP loci of different color of Bletilla striata, the primers were designed for identifying different color of Bletilla striata. Result showed that：two kinds of flower color of Bletilla striata genomic DNA can be amplified based on 4 pairs primers of plant DNA barcode markers and SNP loci were found. None SNP locus was found in trnH-psbA sequence. Two SNP loci were found in rbcL sequence. Sixty-two SNP loci were found in ITS sequence. Three SNP loci were found in LSU D1-D3 sequence. Primer DUAN5（5′-TAGTAACGGCGAGCGAAC-3′） and JDHH（5′-TTATCAGCTTCCTTGCGCCCG-3′） can be used for identifying the yellow flower color of Bletilla striata according to the SNP loci in LSU D1-D3 sequence. The primers were found for identifying the yellow flower color of Bletilla striata.

Key words: Bletilla striata, DNA barcode, PCR, SNP

CLC Number:

R282

ZENG Li-na, LIU Xian-yi, SHI Jian-long, LIANG Da-yong, ZHANG Jie, ZHANG Ting-ting, REN Jian-guo, LIU Hong-mei. Sequence Analysis of Different Flower Color of Bletilla striata Based on Four DNA Barcoding Markers[J]. HUBEI AGRICULTURAL SCIENCES, 2018, 57(17): 106-110.

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