The Study On Genetic Diversity Of Genus Ligulariain Changbai Mountain

the paper on-the-spot investigation and collect five pieceses genus Ligularia, observe them in every detail. use the teconology of paraffin section to observe the structure about the five genus Ligularia. use the teconology of RAPD markers to find the genetic diversity of the 20 pieces material genus Ligularia. it can be provide the reference about Ligularia in molecular biology and paraffin section, in order to reveal the relation-nshape, origin and evolution.1. Morphology and Anatomy Structures of five typs genus Ligularia(1) According to theresult of documentary records specimen adjusting andon-the-spot investigation, formation the key of five pieceses genus. Ligularia from Changbai Mountain.(2) five pieceses genus Ligularia has bifacial leaf that their mesophyll differentiation is obvious, the leaf consists of epidermis, mesophyll and vein, the palisade tissue is well developed. There is someglandular hair in epidermis of the Ligularia jasesii and the Ligularia mongolica; There are some pores in vein of the Ligularia fischeri,the Ligularia intermedia and the Ligularia jaluens; Five kinds of genus Ligularias have a lot of vascular bundle, the typeis bicollateral vascul bundlein mainvein, The vascular bundle of the Ligularia jasesii and the Ligularia mongolica in lateral nerve is very developed.(3) The Major vein diameter,vane thickness and Sponge tissue thickness of the Ligularia jasesii are Significantly less than the Ligularia intermedia,the Ligularia jaluens,Ligularia jasesii and the Ligularia mongolica;The Epidermis thickness of the Ligularia jasesii is Significantly greater than the other four types of Ligularias;the epidermis thickness of the Ligularia fischeri is Significantly greater than the other four types of Ligularias; The Stockade tissue thickness of the Ligularia fischeri is Significantly less than the other four Ligularias.2.The research of genetic diversity by RAPD(1) The genetic DNA of five genus Ligulariause from Changbai Mountain of 20 pieces material can be extracted by the SDS(I),improved CTAB I (II) and improved CTAB II (III). But the quantity, concentration and the purity are different. The sequence of purity from high to low isⅢ> I> II. The concentration isⅢ>Ⅰ> II; the quantity isⅢ> I>Ⅱ. So the improved CTAB(Ⅱ) is the best method(2) Use theLigularia fischeri achieve PCR reaction system for five genus Ligulariause from Changbai Mountain of 20 pieces material was as follows:the optimize system was 20 ng/μLDNA, lOpmol/L primer,1.5Unit Taq polymerase, dNTP300μmol/L, Mg2+1.5mmol/L,10×Buffer 2.5μL in 20 u L reaction system, and the lack part complement with DW. The reaction system was as follows:predenature at 94℃for 5 min, and then 94℃1 min,40℃1 min,72℃1 min30s, for 40 cycles, finally extended at 72℃for 10mins.(3) 12 random primers combinations which combinations polymorphism in five genus Ligularia from Changbai Mountain of 20 pieces material were screened from RAPD primer combination, and 52 diversity strips were obtained, and the polymorphism rate is 71.43%.The genetic distance is 0.08 to 0.49, and on average is 0.29; the minimum genetic distance between materials is 0.08, It appears in the cultivar Ligularia fischeri-1 and the cultivar Ligularia fischeri-2; the maximum distance is 0.49, appears in the cultivar Ligularia fischeri-1 and the Ligularia mongolica-20. so the shorter the distance is, the closer the genetic relationship is.(4) five genus Ligularia from Changbai Mountain of 20 pieces material can be divided into three categories in the level of 0.248 in genetic distance. The first categories is the Ligularia fischeri;the second is the Ligularia intermedia,the Ligularia jaluens and the Ligularia jasesii;the third is the Ligularia mongolica. the Ligularia intermedia and the Ligularia jaluens first clustering, after that the Ligularia jasesii, then the Ligularia mongolica, the Ligularia fischer ilast clustering.