Construction Of A Rapid Screening System For Mushroom Breeding In Pleurotus Nebrodensis

Pleurotus var. nebrodensis is a kind of rare medicinal edible fungi, of which fruit body is white and unique flavor.It is known as high protein,rich nutrition and low in fat.In addition,it is a natural health food which has the efficacy of anticancer.Therefore, nebrodensis is popular with consumers for its economic and ecological values,which has the broad cooperative prospects. However, people have some trouble in cultivating for its special ecological environment and biological characteristics.The malformation of fruitbody and the low rate of mycelia not only lead to the long production cycle but also seriously low rate in biological conversion.The purpose of the present study is to establish an efficient rapid screening system in mononuclear mycelium mating type and good hybrids screening with molecular markers.The screening system could short the time of cross breeding program and meet the demend of rapidly developmentof maket,which can promote the income of peasants and solve the problem of spare labour force.It is importantl to promote the economic benefit of cultivation.The results obtained in the present study were as follows:1. About 39 monokaryon strains were obtained by protoplast monokaryogensis method and microscopic examination. 24 potive myceliums and 15 negative myceliums were obtained by antagonistic action and confirmation by clamp connections. Random Ly selecting 5 myceliums in the positive and negative myceliums was not only for ISSR and SRAP molecular markers with parents, but also developing SCAR.7 specific pairs of primers were obtained including 3 positive primers and 4 negative primers. The result of multiple PCR was right.2. The parent produced 52 myceliums by protoplast monokaryogensis method.Only one possible rejuvenated strian, numbered as hybrid 45×49,was obtained by antagonistic action and confirmation by clamp connections. Biomass, mycelial growth rate, as well as lingo-lignocellulosic degradation enzymes were assayed. Results indicated that there was no significant difference at P<0.01 in mycelial growth rate between the degenerated strain and rejuvenated strain. Biomass and activities of lingo-lignocellulosic degradation enzymes of the rejuvenated strain was higher than the degenerated strain PDB broth. Methods for rapid differentiation of degenerate strains for cultivated P. nebrodensis was established and the decolorizing ratio reached D<60.3. Collecting spores of three kinds nebrodensis random Ly matched. Many hybrids could be obtained. Regard the different hybrids and parental genome DNA as a template for SRAP and ISSR molecular makers. The NTSYS-sp software was not only applied to analyse the result of ISSR and SRAP molecular makers and establish the UPGMA tree but also analyse the genetic relationship between parents and hybrids. Agronomic characters of hybrids were tested by cultivation.Some conclusion were come by analyzing the relationship between the agronomic characters of hybrids and the UPGMA tree, which can guide the future breeding test.