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Application of 2-amino-3-methylhexanoic acid in improving quality of tea leaves

A technology of methylhexanoic acid and amino group, applied in the field of agricultural biological pesticides, can solve the problems of no research, reports and patents related to plant immune induction, no inhibitory effect, no natural microorganisms, etc., and achieves easy artificial synthesis and simple structure. Effect

Active Publication Date: 2021-10-26
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Biological activity research shows that 2-amino-3-methylhexanoic acid has obvious inhibitory effect on Bacillus subtilis and Escherichia coli K-12, and has obvious inhibitory effect on Lactobacillus butyricum (Achromobacter butyri ), Arthrobacter urea (A.ureafaciens), Escherichia coli B (E.coli B) and Pseudomonas aeruginosa (Pseudomonasaeruginosa) have slight inhibitory effect, but to Aerobacter aerogenes (Aerobacter aerogenes), Brevibacterium flavum ( Brevibacterium helvolum), Pseudomonas fluorescens (P.fluorescens) and Serratia marcescens (S.marcescens) had no inhibitory effect
In addition, in 2002, Muramatsu et al. found that E. coli engineering bacteria producing hirudin analogues could synthesize 2-amino-3-methylhexanoic acid, but did not study its activity
So far, there are very few studies on 2-amino-3-methylhexanoic acid, all of which focus on the biosynthetic pathway of the substance synthesized by bacterial mutants or recombinant engineered bacteria (non-natural microorganisms) and the direct inhibition of bacterial activity. The report of microorganisms producing this substance does not involve related research, reports and patents on plant immune induction, which is where the innovation of this patent lies

Method used

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  • Application of 2-amino-3-methylhexanoic acid in improving quality of tea leaves
  • Application of 2-amino-3-methylhexanoic acid in improving quality of tea leaves
  • Application of 2-amino-3-methylhexanoic acid in improving quality of tea leaves

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1: 2-amino-3-methylhexanoic acid induces tobacco resistance to tomato spotted wilt virus infection

[0040] 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. For Bacillus transformation, the transformed Agrobacterium solution was spread on the corresponding resistance screening plate, and cultured at 28°C (±1) for 48h. Pick a single colony of Agrobacterium on the transformation plate, place it in 5 mL of ...

Embodiment 2

[0054] Embodiment 2: 2-amino-3-methylhexanoic acid induces wheat resistance to powdery mildew infection

[0055]2-Amino-3-methylhexanoic acid was dissolved in distilled water and then diluted with distilled water to form 1nM, 10nM, 100nM, 1000nM and 10000nM solutions, and a blank control was set up. After the wheat seeds are germinated, they are planted in sterilized soil cultivation pots, and placed in a greenhouse at 23(±1)°C for 12 hours of light for cultivation. When the seedlings grow to the stage of 1 leaf and 1 heart, spray the stems and leaves of the wheat seedlings with the 2-amino-3-methylhexanoic acid solution of the above concentration, and repeat the treatment once every 24 hours, and carry out two treatments in total, 24h Finally, fresh spores of powdery mildew of wheat were sprinkled evenly on the wheat leaves, and 3 pots were treated with 20 plants per pot. After 10 days, investigate the wheat disease level of each treatment, record the disease degree accordin...

Embodiment 3

[0066] Example 3: 2-amino-3-methylhexanoic acid induces Arabidopsis resistance to Pseudomonas syringae infection

[0067] Dissolve 2-amino-3-methylhexanoic acid in sterile water, and then use sterile water to gradiently dilute it into 1nM, 10nM, 100nM, 1000nM and 10000nM solutions. A blank control is also set up, and 0.02% Tween 20 is added at the same time as Surfactant. Spread Pseudomonas syringae PstDC3000 on an LB plate and culture at 28°C for 48 hours; pick a single colony and inoculate it into a 50mL centrifuge tube containing 2mL of medium, culture it on a shaker at 28°C and 250rpm, monitor every 1-2h Bacteria solution OD 600 value change, at OD 600 Stop culturing bacteria before the value reaches 0.8; transfer 1mL of bacterial liquid to a sterile 1.5mL centrifuge tube, centrifuge at 8000rpm for 2min, and collect the precipitate; remove the supernatant, wash the precipitate with 10mM magnesium chloride for 3 times and centrifuge, and finally resuspend PstDC3000 in 10...

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Abstract

The invention discloses an application of 2-amino-3-methylhexanoic acid in improving the quality of tea leaves. The 2-amino-3-methylhexanoic acid solution treatment can effectively relieve the damage of high temperature to the tea leaves, the treatment effect of 100 nM is optimal, and the heat damage rate of the tea leaves treated by the concentration is obviously lower than that of a control group. And the photosynthetic performance index PIABS of the tea leaves is obviously higher than that of a control group, so that the serious inhibition of high temperature on the photosynthesis of the tea leaves can be effectively relieved by spraying the 2-amino-3-methylhexanoic acid. Meanwhile, the total amino acid content of the tea leaves treated by the 2-amino-3-methylhexanoic acid solutions with different concentrations is obviously higher than that of a control group; under the normal temperature condition, the amino acid content of the tea leaves can also be obviously increased through treatment of 2-amino-3-methylhexanoic acid.

Description

[0001] Case Description [0002] This application is a divisional application of the Chinese invention patent application with the application number 2020115494866, the invention name is "2-amino-3-methylhexanoic acid as a plant immune inducer", and the application date is December 24, 2020. . technical field [0003] The invention belongs to the field of agricultural biological pesticides and relates to the application of 2-amino-3-methylhexanoic acid in improving the quality of tea. Background technique [0004] Plants are constantly infected by various pathogenic microorganisms during their growth and development. Once many diseases occur in agricultural production, they cannot be prevented and controlled by existing agricultural technologies. Therefore, disease prevention is particularly important. At present, the prevention and control of plant diseases mainly adopts the strategy of using pesticides to directly kill pathogenic bacteria, but long-term and large-scale us...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N37/44A01P21/00A01P3/00
CPCA01N37/44A01P15/00A01P21/00A01P1/00A01P3/00A01N25/30
Inventor 陈世国李晶晶汪岚王赫强胜房婉萍张裕
Owner NANJING AGRICULTURAL UNIVERSITY
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