Application Of Chloroplast MatK And RbcL Sequences To Preliminarily Study On Genetic Diversity Of Vaccinium Uliginosum Germplasm Resources

Vaccinium uliginosum is a deciduous shrub or small shrub of Vaccinium in Ericaceae.The contents of anthocyanins and flavonol glycosides in ripe fruit are higher than common fruits and vegetables,and have outstanding antioxidant activity.'In this study,40 Vaccinium uliginosum samples from different wild populations were used as materials to PCR amplification the chloroplast matK and rbcL gene fragments,sequence alignments and haplotype analysis were conducted to investigate the application of two gene fragments for the feasibility of the analysis of genetic diversity of Vaccinium uliginosum,the results show:1.Two pairs of DNA barcode primers were designed and synthesized.40 samples from different wild populations Vaccinium uliginosum were used for PCR amplification.The sequence lengths of 40 samples matK sequence from Vaccinium uliginosum all were 883 bp,and the sequence length of rbcL all were 599 bp,and the homology consistency of the Vaccinium uliginosum obtained from GeneBank reached 99%,indicating that the matk and rbcL gene fragments of Vaccinium uliginosum were successfully cloned.2.Sixteen variable sites and eight parsimony informative sites were found in the 40 matK sequences of Vaccinium uliginosum,which could be divided into 15 haplotypes.The sequence average nucleotide diversity was 0.00357,the haplotype diversity was 0.8551.The Hd value of each population ranged from 0.400 to 1.000.Among them,the haplotype diversity of N population located at DongFangHong Forest Farm of ChangBai Mountain was the highest,with a value of 1.000.H2 haplotypes accounted for 5 samples in a single population were the largest,with a value of 80%,H2 haplotype accounting for 27.5%of the 40 samples in different populations,indicating that the H2 haplotype has a higher proportion within the populations inside and between populations.3.Five variable sites and two parsimony informative sites were found in 40 rbcL sequences of Vaccinium uliginosum,which could be divided into 7 haplotypes.The sequence average nucleotide diversity was 0.00095,the haplotype diversity was 0.465.The Hd value of each population ranged from 0.000 to 0.700.Among them,the haplotype diversity of the S population located at WangQingLanJia Forest Farm is the highest,with a value of 0.700.H1 haplotype accounts for 100%of N population and P population,H1 haplotypes were distributed among 8 populations,and H1 haplotype accounted for 70%of the 40 populations in different populations,indicating that H1 haplotypes all have a higher proportion within the populations inside and between populations.4.For the study of DNA sequences,the genetic diversity of the population is usually measured by the number of haplotypes,nucleotide diversity,and haplotype diversity.Comprehensive analysis shows that the genetic diversity of Vaccinium uliginosum is at a low level.In this experiment,the matK sequence of Vaccinium uliginosum had higher genetic diversity and the rbcL sequence had lower genetic diversity.Therefore,the matK sequence was more suitable for the genetic diversity of Vaccinium uliginosum germplasm resources.