【摘要】：本文針對沙縣金農(nóng)菌業(yè)有限公司提供的金針菇"江山一號"菌株,進行了營養(yǎng)要素的篩選和液體菌種及培養(yǎng)條件的優(yōu)化研究,并完成了液體菌種與常規(guī)固體菌種栽培對比試驗。首先,本文對金針菇"江山一號"菌株的rDNA-ITS序列進行分析,并從GenBank中下載國內(nèi)不同地區(qū)主栽金針菇品種的ITS序列,分析"江山一號"與國內(nèi)主栽金針菇品種的遺傳差異;運用單因素實驗篩選出了最適合金針菇"江山一號"生長的碳氮源、碳氮比以及生長所需的常量元素、微量元素、維生素,再從三種防腐劑中篩選出了能有效促進菌絲生長的雙乙酸鈉,然后采用L9(34)型正交實驗進一步優(yōu)化培養(yǎng)基各組分含量,得到一個適合"江山一號"菌株液體發(fā)酵的營養(yǎng)配方;在得到營養(yǎng)配方的基礎上,以生物量為主要評價指標,通過單因素實驗,對培養(yǎng)溫度、培養(yǎng)時間、接種量、裝瓶量、搖床轉速以及初始pH值進行優(yōu)化,得出了"江山一號"菌株液體菌種的最佳培養(yǎng)條件。最后在沙縣金農(nóng)菌業(yè)有限公司進行初步應用,并和傳統(tǒng)固體菌種生產(chǎn)進行比較。為推廣"江山一號"菌株液體菌種栽培提供參考依據(jù)。主要研究結果如下:(1)"江山一號"菌株與國內(nèi)主栽的金針菇品種都存在一定程度的差異,與其它品種的平均遺傳距離為0.008,最大遺傳距離為0.017。(2)"江山一號"生長營養(yǎng)要素:最適碳源為面粉,其次為玉米粉;最適氮源為黃豆粉,其次為酵母粉;菌絲在碳氮比30:1的培養(yǎng)基生長速度最快;常量元素中,鉀、磷、鈣、鎂離子能顯著地影響金針菇菌絲的生長,其中對金針菇菌絲體生長影響最大的是鉀離子;微量元素中鋅、鐵離子對菌絲生長有促進作用;6種維生素(生物素、核黃素、葉酸、抗壞血酸、硫胺素、尼克酸)對金針菇菌絲體生長影響不顯著;促進劑中濃度為0.02%的雙乙酸鈉能夠極顯著地促進菌絲生長。(3)經(jīng)正交實驗優(yōu)化后,"江山一號"生長最佳營養(yǎng)配方為:玉米粉2%、黃豆粉0.1%、酵母粉0.1%、磷酸二氫鉀0.15%、雙乙酸鈉0.02%。(4)"江山一號"液體菌種最適培養(yǎng)條件為:溫度25℃、裝液量100mL(三角瓶容量250mL)、接種量10%、搖床轉速160r/min、培養(yǎng)周期6 d、起始pH 6.0。(5)"江山一號"液體菌種和固體菌種出菇試驗對比結果:與固體菌種相比,液體菌種菌絲的萌發(fā)時間提前2d左右,菌絲滿袋時間提前10 d左右,子實體現(xiàn)蕾時間提前9 d左右。子實體的平均單產(chǎn)量和生物學效率方面,液體菌種均優(yōu)于固體菌種。
[Abstract]:In this paper, the screening of nutrient elements and optimization of liquid bacteria and culture conditions of "Jiangshan No. 1" strain of Flammulina velutipes provided by Shaxian Jinnong Bacteriological Industry Co., Ltd. were carried out, and a comparative experiment of cultivation of liquid and conventional solid strains was completed. Firstly, the rDNA-ITS sequence of Flammulina velutipes strain "Jiangshan No. 1" was analyzed, and the ITS sequence of Flammulina velutipes cultivated in different regions of China was downloaded from GenBank, and the genetic difference between "Jiangshan No. 1" and domestic Flammulina velutipes was analyzed. The single factor experiment was used to select the most suitable carbon and nitrogen source, C / N ratio and elements, trace elements and vitamins for the growth of Flammulina velutipes "Jiangshan No. 1". Sodium diacetate, which can effectively promote the growth of mycelium, was selected from three preservatives, and then L9 (34) orthogonal experiment was used to further optimize the content of each component in the culture medium, and a nutrient formula suitable for liquid fermentation of "Jiangshan No. 1" strain was obtained. On the basis of the nutritional formula, the biomass was taken as the main evaluation index, and the temperature, time, inoculation amount, bottling quantity, rotating speed of shaking bed and initial pH value were optimized by single factor experiment. The optimum culture conditions of liquid strain of "Jiangshan No. 1" strain were obtained. Finally, preliminary application was carried out in Shaxian Jinnong Bacteriology Co., Ltd, and compared with traditional solid bacteria production. It provides reference basis for popularizing liquid strain cultivation of Jiangshan 1 strain. The main results are as follows: (1) there are some differences between "Jiangshan No. 1" strain and Flammulina velutipes cultivated in China. The average genetic distance with other varieties was 0.008, and the maximum genetic distance was 0.017. (2) the most suitable carbon source was flour, followed by corn flour, and the most suitable nitrogen source was soybean powder and yeast powder. The growth rate of hyphae was the fastest in the medium with 30:1 C / N ratio. Potassium, phosphorus, calcium and magnesium ions in constant elements could significantly affect the growth of Flammulina velutipes hyphae, among which potassium ion was the most important factor in the growth of Flammulina velutipes mycelium, and zinc in trace elements was the most important factor in the growth of Flammulina velutipes mycelia. Iron ions can promote the mycelium growth of Flammulina velutipes. Six vitamins (biotin, riboflavin, folic acid, ascorbic acid, thiamine, nicotinic acid) have no significant effect on the mycelium growth of Flammulina velutipes (Flammulina velutipes). The concentration of 0.02% sodium diacetate in accelerator could significantly promote the growth of hyphae. (3) after orthogonal experiment, the optimum nutrition formula of "Jiangshan No. 1" growth was: corn flour 2, soybean powder 0.1, yeast powder 0.1, potassium dihydrogen phosphate 0.15, diethyldiethylbenzene 0.15. (4) the optimum culture conditions of "Jiangshan No.1" liquid bacteria were as follows: temperature 25 鈩，

本文編號：2245016