| Ramaria botrytis(Pers.)Ricken is an important wild edible fungi in our country,it also has a variety of medicinal functions,therefore,it has a great prospect of development and application.Compared to the common large fungus,R.botiytis is unable to artificially cultivate and fruitingbodies cannot be acquired in large quantities,which is difficult to meet people's needs.This experiment through separated the strain from field collection of fruiting bodies of R.botrytis,study on growth characteristics of isolated strain of R.botiytis,in order to provide evidences for the artificial cultivation of this rare wild edible and medicinal fungi.This thesis has designed the selective medium of basidiomycetes as strain isolation medium of R.botrytis at first,salicylic acid-Fe3+,salicylic acid-Fe3+-MSL,salicylic acid-Fe3+-HED,salicylic acid-Fe3+-MSL-HED were used as antimicrobial agents,respectively,75?g/mL salicylic acid-Fe3+-MSL-HED selective medium of basidiomycetes was selected as the best culture medium.By means of tissue separation,tissue breaking and spore ejection,the tissue blocks at the junctions of the base,middle,stipe and branch of fruitingbody of R.botiytis were investigated in three different medium to studydifference in mycelial germination,the results show that the isolated strain from R.botrytis could germinated on the optimal selective medium of basidiomycetes and the contamination rate was low,the time needed for germination of mycelia isolated by tissue breaking is the shortest,but the pollution rate is high;the germination and growth of mycelium from the junction of the stipe and branch was the fastest.It was identified by macroscopic and microstructural observation and molecular biological methods that the isolated strain numbered 160710 were isolated from the fruitingbody of R.botrytis.The effects of liquid culture conditions on the mycelial growth of isolated strain-160710 from R.botrytis were studied by single factor and orthogonal experiments,respectively.Eight factors including carbon source,nitrogen source,inorganic ion combination,growth factor,initial pH of culture medium,culture temperature,shaking speed and light were tested.The results showed the optimum culture conditions were as follows:at 23? and shaking speed of 220 rpm,initial pH 6.5,flask volume 50 mL/250 mL erlenmeyer flask,inoculum density 10%(v/V),full darkness culture for 7 days,the optimum liquid culture medium for 1L was 25 g soluble starch,3.5g yeast extract,1.2 g KH2PO4,1.6 g FeSO4·7H2O,1.6 g MgSO4·7H2O and 0.01 g VB1,and under the above conditions the mycelial growth of isolated strain-160710 from R.botrytis was 26.8 g/L.Sawdust,branand and rice were allocated to simulate the wild-type cultures to optimize the growth rate of isolated strain-16071 0 from R.botrytis,so as to shorten the time of culture;the grain strain culture material of rice,wheat and bran was designed to prepare the granular strains of isolated strain-160710 from R.botrytis.obtain a large number of high quality strains for the cultivation of R.botrytis;The growth time of isolated strain-160710 from R.botrytis was the shortest in the culture material with corn core as the main raw material,both different culture after-ripening periods,environmental factor stress and soil-covered cultivation were failed to induce the primordium.The strains of R.botrytis were isolated successfully,which provided the conditions for the protection of the germplasm resources and the cultivation of the mushroom in order to develop and utilize the large edible and medicinal fungi. |
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