| Fagopyrm tatarium,belongs to Polygonaceae,Fagopyrum Mill,a kind of dietotherapeutic crop.Tartary buckwheat is widely cultivated in China,and evolved into abundant resource reservation and genetic diversity.Because of the differences of the genotype and the phenotype between the samples from different geographic origin,so it is significant to analysis the genetic diversity.Tartary buckwheat has a high content of flavoind compounds and the PAL is the first enzyme in the flavoinds synthesis pathway.So in this study,PAL gene diversity in tartary buckwheat was studied in order to provide a basis for selecting cultivated variety with high-content flavonoids.According to the Genbank,we have known the sequence of the phenylalanine ammonia-lyase of tartary buckwheat.So in order to accomplish the amplification of the PAL gene,3 pairs of primers were used,then checked and aligned the sequence by using software and finally analyzed the genetic diversity.The results showed that the length of the phenylalanine ammonia lyase gene was 2011 bp.The variable sites was160,while the parsimony informative sites was 33,accounting for 7.9% and 1.64% of the total length,respectively.Totally 46 SNPs were identified in 67 accessions with the frequency of one SNP per 43.7 bp,including 23 base transition and 23 base transversion,and the ratio of them is 1:1.The intra-specific mean distance within the group ranged from 0.002-0.016.The Sichuan group was 0.016,while the tartary buckwheat from the Inner Mongolia group showed that the intra-specific mean distance was 0.002.And it revealed that there is a significant difference among the tartary buckwheat resources in genetics diversity from different geographic origins.While the inter-specific mean between the groups showed that the value between Sichuan and other groups was more than the others.We checked the 67 PAL sequences using DNASP.And it showed that the average value of the Nucleotide polymorphism(?)and the average heterozygosity(?)were0.0034 and 0.0148,respectively.The Nucleotide polymorphism(?)value in samples from Sichuan was the highest(0.0148).And according to the genetic distances and thegenetic diversity parameters,we could concluded that the samples from Sichuan regions had abundant genetic diversity and this may be related with the southwest region of China is the origin of tartary buckwheat.From the phylogenetic tree,it indicated that the 67 samples from the different province could be clustered into seven category.And the 5 sample in Tibet were clustered into another category.These results showed that the PAL sequences of the majority tartary buckwheat samples was stable and the differences between the most samples was not significant.The samples in Sichuan province had abundant genetic diversity.But only in samples from Tibet it revealed many SNPs site,and this should be the new mutation spots in the PAL coding regions.The buckwheat is a kind of crop which contains abundant flavoinds compounds.To compare inhibitory effect of different extracts from buckwheat and some flavonoid compounds on the non-enzymatic glycosylation of protein in vitro.The inhibitory activity of buckwheat extracts and the flavonoid compounds against the formation of advanced glycation end-products(AGEs)was evaluated by the bovine serum albumin-glyoxal(BSA-GO)model.The results indicated that the inhibitory rate of the2#buckwheat ethanol extract is the highest.followed by the the 1# buckwheat ethanol extract,the metformin hydrochloride and the buckwheat water extract.While the buckwheat water extract group presents the lowest inhibitory rate.Both of the ethanol extracts show no significance on the inhibitory rate comparing to the standard flavoinds compounds(i.e.rutin,quecetin and isoquecetin).The buckwheat ethanol extract and the flavonoid compounds could inhibit the non-enzymatic protein-glycated reaction in vitro,the buckwheat might be applied to drug use for the non-enzymatic protein-glycated reaction. |
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