Bacillus methylotrophicus and application thereof
A methylotrophic, bacillus technology, applied in the application, bacteria, fungicides and other directions, to achieve the effects of simple use, short production cycle and low cost
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Embodiment 1
[0041] Rapeseed Sclerotinia, rice sheath blight, watermelon Fusarium wilt, and walnut root rot described in this embodiment are provided by the Plant Pathology Research Office of the Agricultural College of Changjiang University. Rapeseed Sclerotinia and rice sheath blight obtained in other ways , Fusarium wilt of watermelon, root rot of walnut are also applicable to the present invention.
[0042] 1. Isolation, screening and identification of methylotrophic Bacillus XK-4
[0043] Isolation: Vigorously growing rapeseed plants were collected from the rapeseed fields around Jingzhou City, Hubei Province, and the rhizosphere soil adhered to the roots was taken and placed in a 250mL triangular flask. Pack the Sclerotinia sclerotiorum sclerotia collected in the previous growing season into 5cm 2 Size, mesh is in the nylon pouch of 1.5mm2, every bag 10, then nylon bag is buried in the above-mentioned triangular bottle that fills rhizosphere soil. Three weeks later, take out the sc...
Embodiment 2
[0095] Embodiment 2 is basically the same as Embodiment 1, the difference is:
[0096] This embodiment completes the preparation of the methylotrophic bacillus XK-4 inoculum according to the following main steps:
[0097] (1) Inoculate the methylotrophic Bacillus XK-4 on LB solid medium, cultivate at 30°C for 22h, and activate the strain; LB solid medium preparation: 5g of yeast extract, 10g of tryptone, chlorine Sodium chloride 10g, agar powder 20g, NaOH to adjust the pH to 7.1, 121 ℃, 20min autoclave.
[0098] (2) Insert the strain activated in step (1) into LB liquid medium, shake and culture at 155r / min at 30°C for 22h to make seed liquid; LB liquid medium preparation: 5g of yeast extract, 10g of tryptone , sodium chloride 10g, NaOH to adjust the pH to 7.1, 121 ℃, 20min autoclaving.
[0099] (3) Inoculate the seed fermentation broth obtained in step (2) at 5% by volume in LB liquid fermentation medium, and shake and cultivate at 30° C. at 140 r / min for 42 hours.
[0100...
Embodiment 3
[0102] Embodiment 3 is basically the same as Embodiment 1, the difference is:
[0103] This embodiment completes the preparation of the methylotrophic bacillus XK-4 inoculum according to the following main steps:
[0104] (1) Inoculate the methylotrophic Bacillus XK-4 on LB solid medium, cultivate at 30°C for 24h, and activate the strain; LB solid medium preparation: 5g of yeast extract, 10g of tryptone, chlorine Sodium chloride 10g, agar powder 20g, NaOH to adjust the pH to 7.2, 121 ℃, 20min autoclaving.
[0105] (2) Insert the activated bacterial strain in step (1) into LB liquid medium, shake and culture at 165r / min at 30°C for 24h to make seed liquid; LB liquid medium preparation: 5g of yeast extract, 10g of tryptone , sodium chloride 10g, NaOH to adjust the pH to 7.2, 121 ℃, 20min autoclaving.
[0106] (3) Inoculate the seed fermentation broth obtained in step (2) at 5% by volume in LB liquid fermentation medium, and shake and cultivate at 30° C. at 140 r / min for 44 hou...
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