Screening A Core Set Of Informative Primers For Identification Of Panax Ginseng And Its Application

Ginseng(Panax ginseng C.A.Meyer),also known as ‘the elite of the ground’,‘the great grass’,or ‘the yellow ginseng’,is a kind of perennial herbaceous medicinal plant that belongs to the Araliaceae family.The raw material of ginseng breeding is its germplasm resources,its rich diversity is the basis for the sustainable development of ginseng industry.Based on the previous research,with the aim of achieving acceleration and precision of ginseng germplasm resources testing,requiring technical support to ginseng seed industry,further improving and optimizing the ginseng variety validation and variety protection and reducing the abundant work of primers sifting during the process of identification,this paper chooses SSR molecular markers to screen out a set of core primers suitable for the study of ginseng germplasm resources,analyzes the ginseng resources of different regions,different varieties and different preservation methods,and validates the effectiveness of core primers.Conclusions drawn as following:1.On the basis previously,the SSR-PCR reaction system and the amplification program were optimized in this experiment using the technology of molecular markers.18 pairs of SSR core primers that were suitable for ginseng germplasm resources studying had been sifted from 121 pairs SSR primers.They were,G03,G05,PG01,gm129,gm175,gm184,P26,P35,P55,P57,pg29,pg281,pg287,pg668,pg1481,pg1663,GB65,and RS10.All of these 18 pairs of primers showed high polymorphism both on PIC value and genetic diversity value.2.The genetic stability of ginseng materials under different preservation methods was studied by using the 18 pairs of SSR core primers.From the genetic level,the results of polyacrylamide gel electrophoresis and capillary electrophoresis showed that plant tissue culture technique could be used as an effective way of ginseng materials conservation,for there were no significant change in genetic materials between generations at the DNA level.Field material performance between the individual plants is very stable.3.The SSR and RAPD markers were used to explore the germplasm resources of ginseng from fifteen kinds.The genetic similarity coefficient and clustering results obtained by RAPD and SSR markers showed that the genetic relationship was related to the geographical distribution in some way,indicating that the two molecular markers both could reveal the genetic diversity and genetic relationship of the ginseng germplasm resources,the core primers selected in this experiment were effective.4.The chosen 18 pairs of SSR core primers were used to detect seven cultivars(line)from inlands and abroad.Among these,one pair of primers(P26)could amplify specific bands of four breeds,another three pairs of primers could amplify specific bands of three cultivars(line).These three pairs of primers were: G05,pg29,pg1481.The primer gm129 could amplify specific bands of two cultivars(line).The seven cultivars(line)of H,XH,S,M,PX1,PX2 and F2 can be distinguished by the combination of the least three pairs of P26,G05 and pg29.P26-b,P26-c,P26-d,P26-e can distinguish S,PX1,PX2,F2 from other cultivars(line).G05-c can distinguish M from other cultivars(line).Pg29-a can distinguish XF from other cultivars(line).G05-a + pg29-a can distinguish H from other cultivars(line).