Cloning And Analysis Of The Gene Fragments Related To Fructification Of Pleurotus Eryngii Var. Tuoliensis

Pleurotus eryngii var. tuoliensis is popular with consumers for its nutritional andmedicinal values. However, the adverse agronomic characters such as the long growthperiod, the poor developing synchrotron of primordium, and the malformation of fruitbodynot only lead to the reduction in its economic benefit of cultivation but also seriously affectits industry development. Therefore, it is important to breed new varieties with theproperty of that primordium forms early and regularly through a combination of thetraditional and molecular breeding methods. Understanding the molecular mechanism offruitbody formation of P. eryngii var. tuoliensis is vital to molecular breeding, and findingthe genes related to fruitbody formation is the first key step of this study.The difference of gene expression between the two developmental stages of primordiaand mycelia of P. eryngii var. tuoliensis was investigated through mRNA differentialdisplay. Total RNAs were extracted from mycelia and primordia with the method of LiClprecipitation.78primer combinations (3anchor primers and26random primers) wereused in differential display PCR amplification and the differential fragments wereseparated by polyacrylamidegelelectrophoresis (PAGE). The authenticity of the differentialfragments was identified by semi-quantitative RT-PCR. The homology analysis of thetarget genes was carried out by bioinformatics.The results showed that the primer combinations that consisted of H-T11A anchorprimer generated the maximum number of the differential and specific bands.A total of eight cDNAs that expressed differentially between mycelia and primordiawere selected finally. One of them had higher expression in mycelia than in primordia,which shared40%homology with a hypothetical protein, and the other seven cDNAs hadhigher expression in primordia, which shared63%,32%,88%,38%,83%,45%,34%homology with the genes coding for cytochrome P450, glycosyltransferase-related protein,non-catalytic module family EXPN protein, glycoside hydrolase family3, ribosomalprotein L29, homocysteine S-methyltransferase and hypothetical protein, respectively.These differential genes were inferred to involve in several metabolic pathways such as energy metabolism, carbohydrate metabolism, cell wall repair and reconstruction andprotein synthesis. This study played a fundamental role in understanding the mechanism offructification and breeding in edible mushrooms.