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Establishment of a DNA Barcoding Database for Legume and Grass Species Identification

DOI: 10.18805/AF-727    | Article Id: AF-727 | Page : 659-668
Citation :- Establishment of a DNA Barcoding Database for Legume and Grass Species Identification.Legume Research.2022.(45):659-668
Yongqing Li, Zunongjiang A. Bu La, Lijun Cao, Shengguo Zhao m18894036031@163.com
Address : Animal Husbandry Quality Standards Institute, Xinjiang Academy of Animal Sciences, Urumqi, China.
Submitted Date : 12-02-2022
Accepted Date : 3-03-2022

Abstract

Background: DNA barcoding, an emerging approach, is being widely used to accurately and quickly identify species using conserved DNA sequences. 
Methods: Herein we designed seven universal primers for matK (matK1, matK2, matK3 and matK4), rbcL, psbA-trnH and ITS based on their nucleotide sequences in GenBank to amplify 40 species of leguminosae and grass forages. Sequence alignment was performed using MEGA 5.0 and haplotype and mutation sites were analyzed with DnaSP 5.10. PCR amplification efficiency on using the primers designed for psbA-trnH and ITS was relatively low, making these sequences unsuitable for DNA barcoding. Further, we optimized target fragment amplification conditions for all 40 species analyzed in this study. On purifying, sequencing and analysing amplification products, we selected 5′ - and 3′ -end conserved fragments in four marked fragments. 
Result: Sequences of each maker loci showed that there were 12, 17 and 6 haplotypes of matK1, matK2 and matK3, respectively and 13 of rbcL. Based on these haplotypes of matK1, matK2, matK3 and rbcL, we established a DNA barcoding database for 20 forage species.

Keywords

DNA barcoding Forages MatK RbcL Universal sequences

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