1Animal Husbandry Quality Standards Institute, Xinjiang Academy of Animal Sciences, Urumqi, China
2College of Animal Science and Technology, Gansu Agricultural University, Lanzhou, China
*Corresponding Authors: Zunongjiang A. Bu La and Shengguo Zhao, Animal Husbandry Quality Standards Institute, Xinjiang Academy of Animal Sciences, Urumqi, China, Email: m18894036031@163.com
**College of Animal Science and Technology, Gansu Agricultural University, Lanzhou, China. zhaosg@gsau.edu.cn
Online published on 15 July, 2022.
DNA barcoding, an emerging approach, is being widely used to accurately and quickly identify species using conserved DNA sequences.
Herein we designed seven universal primers for matK (matKI, 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.
Sequences of each maker loci showed that there were 12, 17 and 6 haplotypes of matKI, matK2 and matK3, respectively and 13 of rbcL. Based on these haplotypes of matKI, matK2, matK3 and rbcL, we established a DNA barcoding database for 20 forage species.
DNA barcoding, Forages, MatK, RbcL, Universal sequences
