Endophytic fungus J12 for promoting growth of casuarina equisetifolia in low-phosphorus environment
A technology of endophytic fungi and casuarina, which is applied in the field of microorganisms, can solve the problems of phosphorus deficiency in rhizosphere soil and low phosphorus mobility, and achieve the effect of promoting seedling height
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[0017] Example 1 Isolation of endophytic fungi from casuarina
[0018] 1. Main equipment
[0019] Ultra-clean workbench SW-CJ-1FD, constant temperature incubator HH•B11-II, constant temperature culture oscillator zhwy-211B, ten-thousandth balance AR1140, automatic vertical sterilizer LMQ.C-4060, ultrapure water Machine P60-CW etc.
[0020] 2. Main Reagents and Media
[0021] Reagents: 15% sodium hypochlorite, 75% absolute ethanol, primer PAGE 11-59bp OD 1-2, DNA electrophoresis loading buffer, GoodViewTM nucleic acid dye, 2×Tap PCR MasterMix, fungal DNA extraction kit, DNA purification and recovery kit.
[0022] Medium: (1) Modified Martin agar medium: peptone 5.0g, yeast extract powder 2.0g, glucose (C 6 h 12 o 6 •H 2 O) 20.0g, Potassium dihydrogen phosphate (KH 2 PO 4 ) 1.0 g, magnesium sulfate (MgSO 4 •7H 2 O) 0.5g, agar 15.0g, ultrapure water 1000ml, pH 6.2~6.6.
[0023] (2) Improved Martin's liquid medium: peptone 5.0g, yeast extract powder 2.0g, glucose (C 6 ...
Embodiment 2
[0042] Bacterial solution preparation: The screened endophytic fungi (J12) were cultured in potato dextrose agar medium at 28°C for one week, then inserted into 60 mL potato dextrose liquid medium, and cultured in a constant temperature shaker for 72 h (28°C, 160 r min -1 ), and then diluted with 40 mL of sterile water, the concentration of the diluted bacterial solution was 8.75×10 5 cfu / mL, this concentration and dose were used for inoculation in subsequent experiments.
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