The Mechanism Of Stimulation Hyphal Growth And Basidiome Initiation Of Agaricus Bisporus By Pseudomonas Pudita

The involvement of Pseudomonas putidaUW4 in the process of basidiome initiation of the cultivated mushroom Agaricus bisporus was investigated. Pseudomonas putida was shown to be responsible for initiating basidiome morphogenesis. Pseudomonas putida and 1-aminocyclopropane-1-carboxylate deaminase (Acds) mutant with Agricus bisporus hypha are incubated together in this research which Pseudomonas putida was shown to be responsible for initiating basidiome morphogenesis. We try to construct an effective genetic transformation system for transformation Acds gene into Agaricus bisporus to enhance its capacity of decomposing ACC from Agaricus bisporus and the biological efficiency. The results were as follows:1 P.putida UW4 or UW4-AcdS- together inoculated with Agricus bisporus on the plating and in the tube, P.putida were shown to be positively attracted toward exudates of mushroom mycelium and adherence of these bacteria to hyphae was observed. Quantitative adherence studies, in conjunction with electron microscopy, revealed the ability of P.putida cells to adhere rapidly and firmly to hyphae. Differences in the chemotactic and adhesive abilities of P.putida UW4 and UW4-AcdS- colonial forms were observed.2 In order to research the effect of P. putida UW4 and UW4-AcdS- on the formation of sporocarp, we used adding P.putida UW4 or UW4-AcdS- to the sterilization earthing, no sterilization earthing and sterilization earthing as the earthing material. The formation time of Agricus bisporus primordium is earlier and larger which use P.putida UW4 to the sterilization earthing than UW4- to the sterilization earthing as the earthing material.3 A PCR product of 1.07 kb was obtained from the total DNA of P.putida UW4 strain, using oligonucleotide primers designed based on the reported sequence of 1-aminocyclopropane-1-carboxylic acid deaminase gene of P. putida.The product was ligated to PET21a vector and sequenced.4 The expression vector PAN7-Acds, carrying gpd promoter, hygromycin selectable gene, was transferred into intact mycelia of Agricus bisporus by particle bombardment. In summary, Agaricus bisporus have the pathway of ACC to ethylene, and ethylene inhibit the growth of its mycelium and primordia formation. The dominant bacteria of casing is the ACC deaminase bacteria. The ACC deaminase bacteria in casing can promote the formation of fruiting bodies, but the ACC deaminase mutant in casing is no effects. The mechanism may be that ACC deaminase degrades ACC and decrease ethylene synthesis by edible fungi, subsequently relieves the inhibitory action of ethylene to mycelia growth and fruit body development of edible fungi.