Bacillus subtilis strain A97, fungicide, and preparation method and application thereof
The technology of Bacillus subtilis and strains is applied in the field of agricultural microorganisms, which can solve the problems of limited inhibitory effect on disease fungi, drug resistance of pathogenic bacteria and high energy consumption, and achieve the effects of improving international competitiveness, low cost and simple preparation method.
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Embodiment 1
[0033] The screening and separation process of Bacillus subtilis strain A97 of the present invention:
[0034] Bacillus subtilis strain A97 was isolated from the Qinling wild ginkgo tree tissue by the Shaanxi Institute of Microbiology. In 2008, the Shaanxi Institute of Microbiology collected wild ginkgo tissue samples from the Taibai Mountain Nature Reserve. The samples were cleaned with distilled water and then the surface was sterilized. The sterilized branch shears, scalpels and punches were used to remove After surface disinfection, the leaves and bark are punched into small pieces of about 1cm2, and the roots, stems, and branches are cut into small pieces of about 0.5cm to 1cm (cuts are required at both ends). The surface disinfection of the tissue block adopts a three-step method. The surface disinfection method and treatment time are shown in the table below.
[0035]
[0036] The surface-sterilized tissue pieces were ground with a sterile mortar and filtered with 4 layers ...
Embodiment 2
[0051] The specific steps of the preparation method of microbial inocula are as follows:
[0052] Step 1. Strain activation:
[0053] The cryopreserved Bacillus subtilis strain A97 was inoculated on a nutrient agar plate medium for activation culture, and cultured at 33°C for 24 hours; then a single colony was inoculated on nutrient agar slant medium for expansion and culture, at 33°C Cultivate for 24 hours; then wash the bacteria with sterile water, and the obtained eluate is used as the inoculum for seed culture.
[0054] The low temperature mentioned in step 1 refers to the temperature generally used for preservation of bacterial species, and those skilled in the art can know from common knowledge. In this example, the Bacillus subtilis strain A97 was stored at -80°C.
[0055] The pH value of nutrient agar plate medium and nutrient agar slope medium is 7.0~7.4, and the components and their mass percentages are: peptone 1%, beef extract 0.5%, NaCl 0.5%, agar 2%, water 96%, The pre...
Embodiment 3
[0065] The specific steps of the preparation method of microbial inocula are as follows:
[0066] Step 1. Strain activation:
[0067] The cryopreserved Bacillus subtilis strain A97 was inoculated on a nutrient agar plate medium for activation culture, and cultured at 28°C for 20 hours; then a single colony was picked and inoculated on nutrient agar slant medium for expansion culture, at 28°C Cultivate for 20 hours; then wash the bacteria with sterile water, and the eluate is used as the inoculum for seed culture.
[0068] In this example, the Bacillus subtilis strain A97 was stored at -80°C.
[0069] The pH value of nutrient agar plate medium and nutrient agar slope medium is 7.0~7.4, and the components and their mass percentages are: peptone 1%, beef extract 0.5%, NaCl 0.5%, agar 2%, water 96%, The preparation method can refer to the national standard GB / T 4789.28-2003 for preparation.
[0070] Step 2. Preparation of seed liquid:
[0071] Fill two 1L Erlenmeyer flasks with 250mL of se...
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