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Screening method of medicine for treating citrus huanglongbing

A technology for citrus huanglongbing and citrus huanglongbing bacteria, which is applied in the field of plant protection, can solve the problems of slow screening of control agents, etc., and achieve the effects of saving costs and improving screening efficiency

Active Publication Date: 2020-03-27
BEIJING UNION UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, Huanglongbing bacteria cannot be successfully cultured in vitro, so the work of screening control agents is slow

Method used

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  • Screening method of medicine for treating citrus huanglongbing
  • Screening method of medicine for treating citrus huanglongbing
  • Screening method of medicine for treating citrus huanglongbing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 obtains candidate compounds by virtual screening

[0049] Using the Haemophilus influenza (Haemophilus influenza) succinyldiaminopimelate desuccinylase gene to compare the whole genome of Candidatus Liberibacter asiaticus (Las) through the GenBank database, a similar sequence was obtained (GenBank: ACT57255. 1)

[0050] Since the protein structure of Haemophilus influenzae succinyldiaminopimelate desuccinylase has been studied most deeply, its protein sequence is similar to the gene protein of succinyldiaminopimelate desuccinylase gene of H. citri. It reached 41.44%, so it was selected as a template to construct the three-dimensional structure of the succinyldiaminopimelate desuccinylase gene protein of citrus Huanglongbing The enzyme gene protein is a dimeric protein with two active centers;

[0051] Named as succinyldiaminopimelic acid desuccinylase protein (abbreviated as: dapE) of H. citriensis Asiatic species, the amino acid sequence encoded by it is sh...

Embodiment 2

[0070] Example 2 Field Screening Compounds for the Treatment of Citrus Huanglongbing

[0071] Since Huanglongbing bacteria cannot be cultivated in the laboratory, the four candidate compounds with higher scores listed in Table 1 were field tested to determine the antibacterial effect of the drug.

[0072] Purchase the screened four candidate compound molecules from Sigma-Aldrich.

[0073] 1. The candidate compound molecule is prepared with equimolar concentration NaOH to make a medicament for watering citrus huanglongbing diseased trees;

[0074] The preparation method is:

[0075] Weigh 1 molar mass of the candidate compound and 1 molar mass of NaOH solid, add it to a small amount of water, dissolve and set the volume to 1 liter, and then make the mother liquor of the medicament;

[0076] 2. Select 12 diseased fruit trees that have grown for 5 years, and the treatment doses are as follows: 0 (control), treatment 1 (50mM), treatment 2 (100mM), treatment 3 (200mM), watering 1 e...

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Abstract

The invention provides a screening method of a medicine for treating citrus huanglongbing. The method comprises the following steps: 1) performing database comparison to obtain a succinyl diaminoheptanedioic acid desuccinylase gene sequence of Asia species of citrus huanglongbing; 2) carrying out mutation on amino acids at the fifth site and the fifteenth site of the succinyl diaminoheptanedioic acid desuccinylase protein sequence of the Asia species of citrus huanglongbing; 3) constructing a dimer protein with two active centers; 4) scoring molecular docking; 5) screening alternative compoundmolecules; 6) preparing a medicament for transfusion of citrus huanglongbing trees; 7) treating the sick trees; 8) taking phloem tissues of diseased branches, and extracting total DNA; and 9) selecting the alternative compound molecules with the most reduced germ concentration as target compound molecules by a PCR method. According to the screening method of the medicine for treating the citrus huanglongbing, the compound capable of treating the plant huanglongbing is obtained by molecular biological analysis and field screening.

Description

technical field [0001] The invention belongs to the field of plant protection, and relates to a method for screening medicines for plant diseases, in particular to a method for screening medicines for treating citrus huanglongbing. Background technique [0002] Citrus fruit is one of the four most productive fruits in the world, and it is also an important economic crop in southern my country. In recent years, Citrus Huanglongbing (HLB) has caused huge losses to local economies and fruit farmers. [0003] Citrus huanglongbing is the number one killer of the world's citrus industry. The disease can infect almost all citrus cultivars, and there is currently no effective cure. Its pathogen is tentatively determined to be α-Proteobacteria and Liberibacter, which are difficult-to-culture Gram-negative bacteria obligately parasitic on the phloem. [0004] Candidatus Liberibacter asiaticus (Las) is the most harmful pathogen of Huanglongbing. It is a Gram-negative bacterium that p...

Claims

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Application Information

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IPC IPC(8): C12Q1/02C12Q1/06C12Q1/6851C12N9/80G16B15/30A01N43/08A01N43/54A01N43/90A01P1/00C12R1/01
CPCC12Q1/025C12Q1/6851C12N9/80G16B15/30A01N43/08A01N43/54A01N43/90C12Y305/01018G01N2333/195C12Q2531/113
Inventor 黄迎春张元朱培林黄丽莉陈金龙
Owner BEIJING UNION UNIVERSITY
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