Analysis Of ITS Sequence-Polymorphism In Rubus And Breeding Of Superior Lines

Rubus germplasm resources have a wide range of species and distribution.It has great value in medicine and food.DNA molecular markers are widely used in the identification of medicinal plants.We collected different regions of Rubus plants.The botanical characteristics of the collected Rubus germplasm resources were described.In addition,the phylogenetic analysis of the germplasm resources of Rubus was carried out by DNA barcoding.For special Chinese medicinal and edibla homologous Rubus chingii Hu of the genus Rubus,we have carried out the breeding of excellent strains.1.359 Rubus germplasm resources from home and abroad were collected.They were planted in dayangdain experimental field of Anhui Agricultural University-.And detailed botanical characteristics of different Rubus species were recorded in this paper.2.Thirteen Rubus plants were used as materials for internal transcribed spacer(ITS)Sequence amplification by PCR methods.Sequence alignment analysis revealed that the full length of ITS sequences of sixteen Rubus plants ranged from 462 to 464bp.All the 5.8S sequences were 164bp with one variable site and content GC in 54.27%or 54.88%.The sequences of ITS2 is either 254 or 255bp with 13 variable sites,27 parsimony-informative sites and 14 single nucletide polymorphism sites and content GC ranged from 53.73%to 55.12%.Phylogenetic tree analysis based on ITS2 sequence revealed that the germplasm resources collected can be divided into four clusters.Among these specials,the genetic distance between A9,A29 and GH14 of both Rubus chingii Hu is very small divided into cluster I.HH2,W5 and JF9 of both Rubus corchonfolius L.f.divided into cluster ?.And red raspberries,yellow raspberries and black raspberries were divided into ? cluster are Rubus.caesius.L sp..The ? cluster is TW1,GA1,S and GB3 are Rubus coreanus Miq..The ITS2 sequence of thirteen species of Rubus and the peripheral standard were used of species identification.3.Excellent strain were selected of palm leaf Rubus chingii Hu.160 germplasm resources were collected from the mountains of southern Anhui,and nine strains with good characters were selected.Four characteristic compounds of immature fruit,Rutin,Tiliroside,Quercetin 3-O-?-D-glucopyranoside and Quercetin 3-O-?-D-galactopyranoside,were detected by HPLC for some strains with market promotion.It was preliminarily determined that Momei no.12 and no.13 strains had large fruit and high contents of characteristic components,which were the excellent strains for further promotion.