Research On Identifying Auricularia Auricula Strains With RAMP Method

Auricularia auricular is a kind of edible fungi which firstly cultivated in China. The production of Auricularia auricular is the seventh compared to the world production of fungi. Auricularia auricular, which stand a significant place for the export of edible fungi mainly export to Japan, Southeast Asia, Western Europe, North America and so on. With the increase of "breeder authority" consciousness, many countries begin to legislate some institution to protect germplasm resource. During the protective current of germplasm resource, our country should enhance the protection. The key point of identical strains is to exploiting a reliable and stabile method. The research chooses 22 production strains of Auricularia auricular which is widespread at production in Jilin province to identify. The purpose is to exploit the rule of hereditary, suitable method of identifying the strains of Auricularia auricular and the reasons of different strains have the same name. This study can offer some base on correlate research. There are three parts in this thesis.First, exploit a method of extracting high quality genome DNA from Auricularia auricular. Because of much jell in mycelium, which combined with DNA when extracting DNA interfere the precipitation of DNA. The study improved a commonly used SDS method and compared with CTAB method, kit and unimproved SDS method. The results indicated that the improved method could well remove polysaccharide and jell in the mycelium when extracting DNA. Through ultraviolet scanning DNA, we found that the improved method obviously elevate the quality of DNA.Second, study the hereditary stability of Auricularia auricular. The mycelium of edible fungi can easily generate degeneration when applying and conserving. The phenomenon of mycelium degeneration is possible due to the variation of hereditary substance. In order to certify this potential variation whether or not generating different strains, this study use the same strain of Auricularia auricular to continual cultivated for 12 generation, than analysis the distinction between the second generation and the twelve generation with RAMP method. The result showed that inheritance of Auricularia auricular is very stability and the conventional machinery damage will not lead to the variation on strain level.Last, using the method of RAMP to identify 22 strains of Auricularia auricular which is widespread produced in Jilin province. This research analysis 40 pair primers, 21 pair primers of which can identify strains of Auricularia auricular, 9 pair primers of the 21 pair primers show well polymorphism, 3 pair primers show well repeatability and polymorphism which can be used for utility primers on strain identify. We can find the phenomenon that same strain have different name through the analysis of the 22 strains, especially in the breeding by farmers.In this research, we find that the primers suited for identifying different strain of Auricularia auricular with RAMP method and find some identical strains which widespread at production in Jilin province have different names. We further proved that Auricularia auricular do not occur variation at strain level. So the phenomenon of "identical-strain-different-name" is caused by human. We improved the DNA isolation methods that purify the quality of DNA significantly.