Cultivation And Preliminary Analysis Of Transcriptome Differences With And Without Test-tube Fruit Body Primordium On Xerocomus Spadiceus

Boletus is a largest genus in Boletaceae which belongs to Boletales of Agaricomycetes of Basidiomycota. It was widely distributed in temperate regions, subtropical and tropical areas in the world. Porcini, the vast majority of them, symbiose with higher plant root system and form ectotrophic mycorrhiza, were a kind of famous edible fungus deeply loved by people because of their fruiting bodies tasting delicious, with abound nutrition and high medicinal value.Most species of Porcini still could not form fruiting bodies through artificial cultivation in addition to individual saprophytic kind which can domesticated and cultivated under laboratory conditions. At present products on the market totally depend on the wild harvested, but with the destruction of the ecological environment and the excessive development of wild resources, they already can not satisfy the increasing market demand, and is difficult to be utilized sustainably.Xerocomus spadiceus is a typical representative in the genus Boletus, and widely distributed in Yunnan province. In this paper, Xerocomus spadiceus, as the research object, induced successfully and innovatively test tube fruiting body primordium based on pure culture. By high-throughput lllumina sequencing platforms to obtain transcriptome data with and without primordium conditions, based on these datas, to identify candidate genes which may involved in fruiting body formation by differential gene expression analysis, and to reveal the molecular mechanism of test tube fruiting body primordium of Boletus. Those reseach above provides certain theoretical basis for regulation fruiting body development using genetic engineering and scale artificial culture for Boletus and other rare wild edible mycorrhizal fungi in the future.The main research contents and results are as follows:1.Using improved PDA culture medium, Xerocomus spadiceus mycelia and test tube fruiting body primordia were induced successfully and identified by ITS molecular markers, and the results showed that the mycelia and primordia all originated from their fruiting bodies.2. Fruit body、mycelium、and primordium RNA were extracted by OMEGA Fungal RNA KIT R6840-01 and their transcriptome sequencing analysis were carried out. The results showed:to the six samples,48.08G raw data obtained; the percentage of Q30 bases more than 85%; 158488 transcript and 49055 unigenes obtained by software splicing; 34081 annotated Unigenes obtained by blast these unigenes with the sequences of public database NR, Swiss-Prot, GO, COG, KOG, KEGG etc.3.The screening of differentially expressed genes showed that differential genes with and without primordium conditions there were together 689, among which, up-regulated genes 444 and down regulated genes 245. Go classification of differential gene showed that differential genes having relationship with cellular components mainly involved in the cell membrane, extracellular region; with molecular function in catalytic, transporter, structural molecule and electron carrier activity etc.; with biological process rhythms process possessing higher proportion. GOG classification of differential gene showed general functional genes in the majority, amino acid transport and metabolism, carbohydrate transport and metabolism, reproduction, recombination and damage repair in the second. KEGG metabolic pathways classification of differential gene showed,129 different genes, mainly distributed in starch and sucrose metabolism(14), oxidative phosphorylation(10), glycolysis/Gluconeogenesis(7), protein processing in endoplasmic reticulum(7), Spliceosome (7) etc. metabolic pathways.