Gene Clone And Analysis Of MYB Transcript Factors In Purple V-marking White Clover And Tatary Buckwheat

White clove(Trifolium repens L.)is one of the most popular leguminous forages in the world and widely spread in China,which has a lot of advantages including soft stems and leaves,rich leaves,rich in nutrition(especially in protein),good palatability,easy recover after clipping,trampling resistance,good regeneration and so on.As one of the major grass for mixed seeding pasture and ornamental lawn planted in the temperate zone,the white clover will play a more and more important role in the ecological animal husbandry,environmental greening and soil and water conservation in our country.Purple V-marking white clover is a nature mutant we bred after many years’ selecting,it is the first dual-purpose white clover variety for landscaping and feeding,which not only enrich the resource bank of white clover in China,but also creating considerable economic value in production,Meanwhile,it also provides new research materials for studying the molecular mechanism of leaf color mutation,lays a solid foundation for further research.Buckwheat is native to China,it is an ancient annual or perennial crop belonging to the family Polygonaceae,genus Fagopyrum,which can be used for food,medicine and feeding.Buckwheat has wide adaptability and high barren resistance,easy to manage and suitable for interplanting,so buckwheat planting has become an important means to improve production and income for farmers.Compared to the main crops such as rice,corn,wheat and soybean,buckwheat is well known for its rich content of rutin(belong to flavonoids)and other active substances and balanced nutrients.Therefore,further illustrate of the molecular mechanism of rutin synthesis in buckwheat may provide genetic resources and theoretical support for high rutin content buckwheat breeding.Transcription factors regulate the expression of anthocyanin biosynthetic pathway by binding with corresponding cis acting elements in promoters of key enzyme genes.MYB is the most widely studied regulatory factor in the regulation of anthocyanin biosynthesis.In this study,a candidate MYB factor TrMYB44 was screened from the transcriptome data both in purple V-marking white clover and normal white clover,and the TrMYB44 gene was cloned,the total length of the gene was 1041 bp and 346 amino acids were encoded.qRT-PCR was used to measure the different expression levels of Tr MYB44 in different organs of purple V-marking white clove,the results show that the leaves had the highest expression level,and the level of purple part of leaves were lower than that of non-purple part,so we indicated that TrMYB44 inhabited the anthocyanin metabolism in leaves.Meanwhile,the plant expression vector pCAMBIA1307-TrMYB44 was successfully constructed,and the expression vector was transferred into the hairy root of Tartary Buckwheat using Agrobacterium tumefaciens infection method,and the genetic transformation was analyzed.What’s more,we cloned 4 MYB transcription factors gene FtMYB13,FtMYB14,FtMYB15,FtMYB16 from Tartary buckwheat,qRT-PCR was used to measure the different expression level of the 4 gene in different organs of Tartary buckwheat,the results show that the 4 gene has different expression mode in different organs.