Production method of nonpathogenic fusarium oxysporum-containing plant vaccine preparation

Abstract

The invention provides a production method of a nonpathogenic fusarium oxysporum-containing plant vaccine preparation. The method comprises the following steps: preparing liquid strain for production by taking nonpathogenic fusarium oxysporum FJAT-9290 as strain and using a potato dextrose agar (PDA) culture medium and a Czapek yeast culture solution as a culture medium for the liquid strain; preparing a solid culture medium substrate for plant vaccine production by taking pig manure waste as a main component, mixing the prepared solid culture medium substrate with bran and wheat grains, respectively packaging the mixture into polypropylene plastic bags, sterilizing and then inoculating the liquid strain, culturing and fermenting for 18-20 days at the constant temperature of 28-30 DEG C to prepare a solid fermentation product of a plant vaccine, and evenly mixing the product after fermentation to obtain the nonpathogenic fusarium oxysporum-containing plant vaccine. The plant vaccine preparation prepared by the method has effects of promoting growth of crops and resisting diseases, can solve the problem that tomato fusarium wilt is difficult to control, and achieves the aim of reducing the use of chemical pesticides.

Description

Technical field

[0001] The invention relates to a production method of a non-pathogenic Fusarium oxysporum plant vaccine preparation.

Background technique

[0002] Crop wilt caused by Fusarium oxysporum is a worldwide soil-borne disease and one of the main causes of continuous cropping obstacles (Liu Bo et al., 2013). Generally, it causes 10%-30% yield loss, and severe cases can lead to no harvest. Timely and effective prevention and control has become a major demand for safe production of Solanaceae and melon crops (Kaur et al., 2010; Yang Yingying et al., 2012). At present, the main strategies for controlling Fusarium wilt are chemical control and cultivating disease-resistant varieties. Chemical control pollutes the environment and endangers human health. Breeding resistant varieties is difficult and there is a risk of weakening resistance. Therefore, it is necessary to open up more control methods. In recent years, the use of plant immune systems to develop safe, ef...

Examples

preparation example Construction

[0032] Step (1) Preparation of liquid strains for production: use non-pathogenic Fusarium oxysporum FJAT-9290 (Fusariumoxysporum) as strains, and use potato dextrose agar PDA medium and Saccharomyces chabzie nutrient solution as liquid strains for cultivation base, to prepare liquid strains for production;

[0033] Step (2) Preparation of non-pathogenic Fusarium oxysporum plant vaccine preparation: the solid medium for plant vaccine production is prepared with pig manure waste as the main component, mixed with bran and wheat grains, and then packed in polypropylene In a plastic bag, after sterilization, inoculate the liquid strain prepared in step (1), culture and ferment at a constant temperature of 28-30°C for 18-20 days to prepare a solid fermentation product of plant vaccine, and mix it well after the fermentation to obtain non-toxic Pathogenic Fusarium oxysporum plant vaccines;

[0034] The Fusarium oxysporum FJAT-9290 (Fusariumoxysporum) was preserved on March 29, 2012 ...

Embodiment 1

[0065] (1) Preparation of liquid strains for production:

[0066] A. Activation of strains: Potato dextrose agar (PDA) medium was prepared, sterilized at 121°C for 20 minutes, cooled to 45-50°C and poured into a sterilized plate for later use. Take the non-pathogenic Fusarium oxysporum FJAT-9290 (Fusarium oxysporum) strain stored at -70°C and culture it by streaking on a PDA plate, and culture at a constant temperature of 28-30°C for 5-7 days;

[0067] The PDA medium: weigh 200g after peeling the potatoes, cut into small pieces of 1cm×1cm, boil for 30min, add 20g of sucrose and 17-20g of agar after filtering through eight layers of gauze, the pH is natural, fully dissolve and set the volume to 1000ml , according to the bottled volume of 200ml / 250ml, sub-package in Erlenmeyer flasks for sterilization;

[0068] B. Inoculate the mycelium in the picking step A in the potato glucose liquid medium, cultivate for 4 to 5 days at 28 to 30° C., and 170 to 200 rpm on a shaker;

[0069]...

Embodiment 2

[0079] (1) with embodiment 1 step (1)

[0080] (2) Utilize the same pig manure waste of embodiment 1 as main culture substrate to produce plant vaccine preparation:

[0081] C. mix by the pig manure waste of 900g and the bran of 100g as basal medium;

[0082] D. Add 400g of wheat grains, 30g of sucrose and 2g of sodium nitrate for every 1000g of basal medium, stir well, add appropriate amount of water, adjust the water volume ratio to 50%, transfer it to a polypropylene plastic bag, and sterilize ;

[0083] The wheat grains need to be soaked for 12-24 hours, so that the wheat grains can fully absorb water and swell.

[0084] F. After the culture medium is cooled, inoculate 5 volumes of the seed liquid (liquid strain) prepared in step (1), and ferment for 18-20 days at a constant temperature of 28 to 30°C, then the fermentation can be completed, and the fermentation product can be stirred evenly to obtain Non-pathogenic Fusarium oxysporum plant vaccine formulations.

[0085...

Technical field

[0001] The invention relates to a production method of a non-pathogenic Fusarium oxysporum plant vaccine preparation.

Background technique

[0002] Crop wilt caused by Fusarium oxysporum is a worldwide soil-borne disease and one of the main causes of continuous cropping obstacles (Liu Bo et al., 2013). Generally, it causes 10%-30% yield loss, and severe cases can lead to no harvest. Timely and effective prevention and control has become a major demand for safe production of Solanaceae and melon crops (Kaur et al., 2010; Yang Yingying et al., 2012). At present, the main strategies for controlling Fusarium wilt are chemical control and cultivating disease-resistant varieties. Chemical control pollutes the environment and endangers human health. Breeding resistant varieties is difficult and there is a risk of weakening resistance. Therefore, it is necessary to open up more control methods. In recent years, the use of plant immune systems to develop safe, ef...

Examples

preparation example Construction

[0032] Step (1) Preparation of liquid strains for production: use non-pathogenic Fusarium oxysporum FJAT-9290 (Fusariumoxysporum) as strains, and use potato dextrose agar PDA medium and Saccharomyces chabzie nutrient solution as liquid strains for cultivation base, to prepare liquid strains for production;

[0033] Step (2) Preparation of non-pathogenic Fusarium oxysporum plant vaccine preparation: the solid medium for plant vaccine production is prepared with pig manure waste as the main component, mixed with bran and wheat grains, and then packed in polypropylene In a plastic bag, after sterilization, inoculate the liquid strain prepared in step (1), culture and ferment at a constant temperature of 28-30°C for 18-20 days to prepare a solid fermentation product of plant vaccine, and mix it well after the fermentation to obtain non-toxic Pathogenic Fusarium oxysporum plant vaccines;

[0034] The Fusarium oxysporum FJAT-9290 (Fusariumoxysporum) was preserved on March 29, 2012 ...

Embodiment 1

[0065] (1) Preparation of liquid strains for production:

[0066] A. Activation of strains: Potato dextrose agar (PDA) medium was prepared, sterilized at 121°C for 20 minutes, cooled to 45-50°C and poured into a sterilized plate for later use. Take the non-pathogenic Fusarium oxysporum FJAT-9290 (Fusarium oxysporum) strain stored at -70°C and culture it by streaking on a PDA plate, and culture at a constant temperature of 28-30°C for 5-7 days;

[0067] The PDA medium: weigh 200g after peeling the potatoes, cut into small pieces of 1cm×1cm, boil for 30min, add 20g of sucrose and 17-20g of agar after filtering through eight layers of gauze, the pH is natural, fully dissolve and set the volume to 1000ml , according to the bottled volume of 200ml / 250ml, sub-package in Erlenmeyer flasks for sterilization;

[0068] B. Inoculate the mycelium in the picking step A in the potato glucose liquid medium, cultivate for 4 to 5 days at 28 to 30° C., and 170 to 200 rpm on a shaker;

[0069]...

Embodiment 2

[0079] (1) with embodiment 1 step (1)

[0080] (2) Utilize the same pig manure waste of embodiment 1 as main culture substrate to produce plant vaccine preparation:

[0081] C. mix by the pig manure waste of 900g and the bran of 100g as basal medium;

[0082] D. Add 400g of wheat grains, 30g of sucrose and 2g of sodium nitrate for every 1000g of basal medium, stir well, add appropriate amount of water, adjust the water volume ratio to 50%, transfer it to a polypropylene plastic bag, and sterilize ;

[0083] The wheat grains need to be soaked for 12-24 hours, so that the wheat grains can fully absorb water and swell.

[0084] F. After the culture medium is cooled, inoculate 5 volumes of the seed liquid (liquid strain) prepared in step (1), and ferment for 18-20 days at a constant temperature of 28 to 30°C, then the fermentation can be completed, and the fermentation product can be stirred evenly to obtain Non-pathogenic Fusarium oxysporum plant vaccine formulations.

[0085...

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