Penicillium composition with low-temperature protection effect, low-temperature protective agent, application of low-temperature protective agent and method for improving survival rate of floating seedlings
A cryoprotectant and cryoprotection technology, which are applied in the fields of botanical equipment and methods, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of food safety, environmental pollution, complicated operation, etc., and achieve low cost and safety. No pollution, simple operation effect
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[0028] In the present invention, the preparation method of the metabolite preferably includes the following steps:
[0029] respectively activating and culturing Penicillium citrinum, Penicillium rubrum, Penicillium Thomme and Paecilomyces variotii to obtain the bacterium liquid of each Penicillium;
[0030] The low-temperature protective agent is obtained by sequentially performing high-temperature inactivation, drying and pulverization on the bacterial agent obtained above.
[0031] In the present invention, Penicillium citrinum, Penicillium rubrum, Penicillium thomerii and Paecilomyces variotii are respectively activated and then cultivated to obtain bacterial liquid of each Penicillium.
[0032] In the present invention, when activating Penicillium citrinum, it preferably includes sterile water activation and culture medium activation. In the present invention, the process of said sterile water activation is preferably to scrape the bacterium colony of Penicillium citrinu...
Embodiment 1
[0049] (1) Prepare medium:
[0050] Cha's medium (Penicillium citrinum): 30.0 g of sucrose, NaNO 3 3.0g, MgSO 4 ·7H 2 O 0.5g, KCl 0.5g, FeSO 4 4H 2 O 0.01g, K 2 HPO 4 1.0g, dilute to 1.0L with distilled water, pH 7.0-7.2;
[0051] PDA medium (Penicillium rubrum): 200g of potatoes, 20g of glucose, dilute to 1000mL with deionized water;
[0052] Malt extract medium (Penicillium Thomme): malt extract 20.0g, dilute to 1.0L with tap water, natural pH;
[0053] 5°Bé wort culture medium (Paecilomyces variotii): 1.0L of 5°Bé wort, natural pH.
[0054] Put the medium prepared above into an autoclave to sterilize at 121°C for 20 minutes;
[0055] (2) Activation and cultivation: Scrape the colony of Penicillium rubrum (just pick a small colony with a bamboo stick) and inoculate it in a 250mL triangular flask filled with 100mL sterile water, at a temperature of 25°C and a rotation speed of 120r Under the condition of 30 min / min, vibrate for 30 min to fully disperse the spore mas...
Embodiment 2
[0062] The application method of the low-temperature protective agent for tobacco seedling cultivation in tobacco seedling cultivation:
[0063] (1) The cryoprotectant I prepared in Example 1 is added to the nutrient solution of floating seedlings at a concentration of 15.00g / kg, and the fungicide (80% carbendazim wettable powder, Sichuan Jiade Li Technology Development Co., Ltd.) is added to the nutrient solution of floating seedlings at a concentration of 100 mg / L to obtain a mixed solution; the nutrient solution is an ammoniated potassium sulfate compound fertilizer solution, and the ammonified potassium sulfate compound fertilizer solution is used to compound potassium sulfate. The concentration of fertilizer is 20g / L (Anhui Zhongyuan Chemical Fertilizer Co., Ltd.);
[0064] (2) Tobacco seeds are placed in the floating polystyrene seedling tray, a total of 84 seeds are placed in the water tank containing the above-mentioned mixed solution subsequently;
[0065] (3) Cultur...
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