Paenibacillus polymyxa and application thereof
A polymyx spore-like and bacillus technology is applied in the field of plant disease biological control to achieve the effects of short production cycle, simple use and broad application prospects
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Embodiment 1
[0041] Rice sheath blight fungus, watermelon wilt fungus, and walnut root rot fungus described in this embodiment are provided by the Plant Pathology Laboratory of Agricultural College, Changjiang University, and rice sheath blight fungus, watermelon wilt fungus, and walnut root rot fungus obtained in other ways are also applicable to the present invention.
[0042] 1. Isolation, screening and identification of Paenibacillus polymyxa WK-3
[0043] Separation: Collect vigorously growing rice plants from paddy fields at the tillering stage around Jingzhou City, Hubei Province, take 15 cm stalks near the roots, wash them with clean water, cut them into small pieces of about 1 cm, place them in sterilized petri dishes, and use Soak in 75% alcohol for 1min, then soak in 0.1% mercuric chloride for 30s, then rinse with sterile water for 4 times, cut the stem longitudinally with sterilized scissors, place in LB liquid medium at 28-30℃, shake at 160r / min Cultivate for 2-3 days, take 1...
Embodiment 2
[0089] Embodiment 2 is basically the same as Embodiment 1, the difference is:
[0090] The present embodiment completes the preparation of Paenibacillus polymyxa WK-3 bacterial agent according to the following main steps:
[0091] (1) Inoculate the Paenibacillus polymyxa WK-3 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, sodium chloride 10g, 18g agar powder, adjust the pH to 7.2 with NaOH, and autoclave at 121°C for 20min.
[0092] (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.
[0093] (3) Inoculate the seed fermentation broth obtained in step (2) at 5% by volume in LB liquid fermentation medium, shake and culture at 160r / min at 30°C for 46h, a...
Embodiment 3
[0097] Embodiment 3 is basically the same as Embodiment 1, the difference is:
[0098] The present embodiment completes the preparation of Paenibacillus polymyxa WK-3 bacterial agent according to the following main steps:
[0099] (1) Inoculate the Paenibacillus polymyxa WK-3 on LB solid medium, cultivate at 29°C for 23h, and activate the strain; LB solid medium preparation: 5g of yeast extract, 10g of tryptone, sodium chloride 10g, 18g agar powder, adjust the pH to 7.1 with NaOH, and autoclave at 121°C for 20min.
[0100] (2) Insert the strain activated in step (1) into LB liquid medium, shake and culture at 155r / min at 29°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.
[0101] (3) Inoculate the 5% seed fermentation liquid obtained in step (2) into LB liquid fermentation medium, shake and culture at 160r / min at 29°C for 47h, and then centrifuge a...
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