Antagonistic bacteria of radix pseudostellariae root rot and application thereof
A technology of antagonistic bacteria and heterophylla, applied in the field of bacterial strains and antagonistic bacteria, can solve the problems of farmland ecosystem function degradation, aggravate environmental pollution, increase production cost input, etc., and achieve good rhizosphere colonization effect and strong cellulose degradation ability, the effect of strong phosphorus-dissolving ability
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Embodiment 1
[0022] Example 1 Screening and identification of antagonistic bacteria Burkholderia ambifaria LK-P4
[0023] 1. Screening of antagonistic bacteria
[0024] The antagonistic bacteria Burkholderia ambifaria LK-P4 of the present invention is screened from the rhizosphere soil of Pseudostellaria heterophylla.
[0025] 1.1LB medium preparation
[0026] Weigh 5g LB dry powder (Haibo Biotechnology Co., Ltd., China) and 3g agar powder (BIOSHARP, Japan), heat and dissolve with deionized water and dilute to 200mL, autoclave at 115°C for 20min, and wait until the temperature of the medium drops to 55°C Add natamycin (fungal antibiotic) when the temperature is around ℃ (touchable by hand) to prevent the antibiotic from becoming ineffective due to high temperature, shake well and pour the plate quickly.
[0027] 1.2 Rhizosphere soil bacterial culture
[0028] Weigh 10g of heterophylla rhizosphere soil, dissolve it in 90mL of sterile water, shake it well, and get 10 -1 Diluent. Take 10...
Embodiment 2
[0037] Antibacterial effect detection of embodiment 2 antagonistic bacteria
[0038] Use PDA medium, inoculate activated Burkholderia LK-P4 at a distance of 2.5 cm from the center of the petri dish, place it in a constant temperature incubator at 28°C for 48 hours, and inoculate each Pathogen-like fungi were placed in a constant temperature incubator at 28°C in the dark for several days, and the formation of the inhibition zone was observed in real time. After 4 days of culture, it was found that Burkholderia ambifaria LK-P4 could effectively antagonize the Fusarium oxysporum specialized in heterophylla, Fusarium moniliforme and Fusarium moniliforme, and inhibit the mycelium growth (such as image 3 shown).
Embodiment 3
[0039] Example 3 Optimization of the optimal medium for Burkholderia ambifaria LK-P4
[0040] Prepare the following medium to optimize the optimal growth medium for Burkholderia LK-P4. The medium is as follows: 1) full LB solution; 2) 1 / 4LB solution; 3) 1 / 4LB+1 / 20MS; 4) 1 / 4LB+1 / 40MS; 5) 1 / 4LB+1 / 80MS; 6) 1 / 4LB+1 / 20MS+1wt.% brown sugar; 7) 1 / 4LB+1 / 40MS+1wt.% brown sugar; 8) 1 / 4LB+1 / 80MS+1wt.% brown sugar.
[0041] Among them, the formula of MS medium (no iron salts) is: 1) macroelements: 1900mg / LKNO 3 , 1650mg / LNH 4 NO 3 , 370mg / LMgSO 4 ·7H 2 O. 170mg / LKH 2 PO 4 , 440mg / LCaCl 2 2H 2 O; 2) Trace elements: 22.3mg / LMnSO 4 4H 2 O, 8.6mg / LZnSO 4 ·7H 2 O, 6.2mg / LH 3 BO 3 , 0.83mg / LKI, 0.25mg / LNa 2 MoO 4 ·7H 2 O, 0.025mg / LCuSO 4 ·5H 2 O, 0.025mg / LCoCl, 2.6mg / LH 2 O; 3) Organic matter: 2.0mg / L glycine, 0.5mg / L pyridoxine hydrochloride, 0.1mg / L ammonium sulfate hydrochloride, 0.5mg / L niacin, 100mg / L creatine; LB medium formula is: pancreas Peptone 10g / L, yeast extra...
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