Application of limonene in preparation of plant resistance inducer
A plant induction and limonene technology, applied in plant growth regulators, plant growth regulators, applications, etc., can solve the problems of disease resistance activity and induced plant stress resistance activity without relevant research reports
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Embodiment 1
[0036] Embodiment 1: Determination of limonene anti-TMV activity
[0037] (1) Protective activity
[0038] Prepare limonene solutions with a concentration of 0.02mg / mL, 0.1mg / mL and 0.5mg / mL, select healthy tobacco leaves at the 5-6 leaf stage with consistent growth, and inoculate 2000-fold diluted TMV solution 48 hours after spraying the drug, and blank control For clear water treatment, the positive control is 0.1mg / mL chitosan oligosaccharide. Each treatment was inoculated with 3 leaves and repeated 3 times. After 3 days, the number of dead spots was counted to calculate the inhibition rate.
[0039] (2) in vitro passivation activity
[0040] Prepare limonene solutions with concentrations of 0.02mg / mL, 0.1mg / mL and 0.5mg / mL, and mix them with equal volumes of 1000-fold diluted TMV solution. Place it at room temperature for 1 hour and inoculate it in the same-growing, healthy 5-6-leaf stage tobacco leaves. The blank control was water treatment, and the positive control w...
Embodiment 2
[0049] Embodiment 2: Determination of limonene-induced tobacco anti-TMV activity
[0050] The heart leaf tobacco was selected to test the induced disease resistance activity. The heart leaf tobacco can form virus scabs, and the symptom on the common tobacco is mosaic. Different symptoms use different methods to count the disease resistance. Spray 0.02 mg / mL, 0.1 mg / mL and 0.5 mg / mL limonene to the lower three leaves of tobacco at the 6-7 leaf stage with consistent growth. After 48 hours, TMV was inoculated to the unsprayed upper leaves. The blank control was water treatment, and the positive control was chitosan oligosaccharide solution. Each plant was inoculated with 2-3 leaves, each treatment included 10 tobacco plants, and the whole experiment was repeated 3 times. After 3 days, the disease index of the number of dead spots of Nicotiana cordata was counted, the formula for calculating the inhibition rate was as above, and the formula for calculating the control effect was...
Embodiment 3
[0058] Example 3: Limonene causes changes in the activity of tobacco defense enzyme POD
[0059] After spraying 0.02mg / mL, 0.1mg / mL and 0.5mg / mL LD-limonene and L-limonene to the leaves, the leaves were taken on the 1st, 3rd, 5th, 7th, 9th, and 11th days, and the defense enzymes PAL and POD were detected. Activity changes. Weigh 5 g of plant leaves, cut them into pieces and place them in a frozen mortar, add a small amount of quartz sand, and add a total of 5 mL of 0.1 mol / L, pH 5.5 acetic acid-sodium acetate buffer in 2 to 3 times. After being ground into a homogenous slurry, centrifuge at 12,000 r / min for 15 minutes at 4°C, and the supernatant is the crude enzyme extract. Take the test tube and add 3mL 25mmol / L guaiacol and 5mL enzyme extract, then add 200μL, 5mol / L H 2 o 2 The solution mixes rapidly to initiate the reaction. Using distilled water as a reference, start to record the value of the reaction system at a wavelength of 470nm at 15s of reaction, record once eve...
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