Application of 2-amino-3-methylhexanoic acid as plant immune inducer
An immune inducer, methylhexanoic acid technology, applied in the field of agricultural biological pesticides, can solve the problems of no inhibitory effect, no natural microorganisms, activity research, etc., and achieve the effect of simple structure
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Embodiment 1
[0042] Example 1: 2-amino-3-methylhexanoic acid induces tobacco resistance to tomato spotted wilt virus infection
[0043] Tomato spotted wilt virus was obtained from Yunnan Province, China. The initial virus source was stored in a -80°C refrigerator. The virus was inoculated on the leaves of Nicotiana benthamiana by friction inoculation to activate the virus, and the virus plasmid was extracted using Escherichia coli competent cells. Transformation, smeared on resistant plates and cultured, picked a single colony for PCR screening, selected positive colonies for sequencing and subsequent plasmid extraction, added the plasmids with normal sequencing to Agrobacterium competent cells, and carried out agronomy by electroshock method. Bacilli were transformed, and the transformed Agrobacterium liquid was spread on the corresponding resistance screening plate, and cultured at 28 °C (±1) for 48 h. Pick a single colony of Agrobacterium on the transformation plate, place it in 5 mL of...
Embodiment 2
[0055] Embodiment 2: 2-amino-3-methylhexanoic acid induces wheat resistance to powdery mildew infection
[0056]2-Amino-3-methylhexanoic acid was dissolved in distilled water and then diluted with distilled water to form 1 nM, 10 nM, 100 nM, 1000 nM and 10000 nM solutions, and a blank control was set up. After the wheat seeds were germinated, they were planted in sterilized soil cultivation pots, and placed in a greenhouse at 23 (±1) °C for 12 h under light for cultivation. When the seedlings grow to the stage of 1 leaf and 1 heart, the 2-amino-3-methylhexanoic acid solution of the above concentration is used to spray the stems and leaves of the wheat seedlings, and the treatment is repeated once every 24 hours, and the treatment is carried out twice, 24 hours After h, fresh spores of powdery mildew of wheat are evenly sprinkled on the wheat blades, and each handles 3 pots, 20 strains per pot. After 10 days, investigate the wheat disease level of each treatment, record the di...
Embodiment 3
[0065] Example 3: 2-amino-3-methylhexanoic acid induces Arabidopsis resistance to Pseudomonas syringae infection
[0066] Take 2-amino-3-methylhexanoic acid dissolved in sterile water and then use sterile water to gradiently dilute to 1 nM, 10 nM, 100 nM, 1000 nM and 10000 nM solutions, set up a blank control, and add 0.02% Tween 20 was used as a surfactant. Spread Pseudomonas syringae PstDC3000 on an LB plate and culture at 28 °C for 48 h; pick a single colony and inoculate it into a 50 mL centrifuge tube containing 2 mL of culture medium, and culture it on a shaker at 28 °C at 250 rpm. -2 h to monitor the OD of the bacterial solution 600 value change, at OD 600 Stop culturing bacteria before the value reaches 0.8; transfer 1 mL of bacterial liquid to a sterile 1.5 mL centrifuge tube, centrifuge at 8000 rpm for 2 min, and collect the precipitate; remove the supernatant, wash the precipitate with 10 mM magnesium chloride for 3 times and centrifuge, and finally PstDC3000 was...
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