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Method and application of compound microorganism treatment to control phloem bacillus pathogen in seedlings

A compound microbial agent and compound microbial technology, applied in the field of citrus virus-free seedling breeding, to improve the efficiency of detoxification and solve the bottleneck problem

Active Publication Date: 2021-09-28
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Since there is still no very effective control method for citrus huanglongbing, cutting down diseased trees, killing psyllids to control the vector and planting healthy and virus-free citrus seedlings are usually used for prevention and control. Among them, the application of virus-free seedlings is very important. One of the countermeasures, therefore, all the excellent varieties that are constantly selected and bred need to be detoxified, and then popularized and applied

Method used

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  • Method and application of compound microorganism treatment to control phloem bacillus pathogen in seedlings
  • Method and application of compound microorganism treatment to control phloem bacillus pathogen in seedlings
  • Method and application of compound microorganism treatment to control phloem bacillus pathogen in seedlings

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Effect test

Embodiment 1

[0040] Example 1 strain identification

[0041] The microbial strains involved in this application were provided by the State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University. In this example, through 16S rDNA amplification, construction of expression vectors and sequencing, it was found that strain TD2-1 had a high degree of homogeneity with Bacillus Velez. Origin, TD2-2 has a high degree of homology with Bacillus amyloliquefaciens.

[0042] 1. Molecular biological identification of strains

[0043] 1) Extraction of bacterial genomic DNA

[0044] After the activation of the strain, take 2ml of the bacterial liquid and centrifuge to collect the bacterial cells, add 350μl of TE buffer solution containing 10mg / ml lysozyme to the pellet, mix gently, and incubate at 37°C for 30min. Then refer to the CTAB method to extract bacterial genomic DNA.

[0045] 2) Amplification of the 16S rDNA fragment of the strain

[0046] Using bacterial genomic DNA as ...

Embodiment 2

[0066] Example 2 Study on the inhibitory effect of compound bacterial agent treatment on the pathogenic source of citrus Huanglongbing

[0067] Preparation of microbial agent by mixed fermentation: pick single colonies of TD2-1 and TD2-2, respectively in LB medium, 200rpm, 37 ° C culture, the OD 600nm =1 Equal amount of TD2-1 and TD2-2 respectively get 10ml and add 20g brown sugar, add in autoclaved sterilized wheat bran (1KG)+rice bran (1KG)+corn starch (0.4KG)+soybean meal (0.2KG), After mixing according to the material-to-water ratio of 1:1, solid fermentation at 30°C for 7 days, during which, the culture was stirred once a day, and the culture was crushed. After drying at 60°C, the culture was crushed into fine powder with a pulverizer to detect live bacteria. The number is 20 billion / g, and it is ready for use. When used, dissolve in water according to the required dosage, 0.5L per plant.

[0068] 1. Bacteria treatment plan

[0069] Shatangju and Nanfeng tangerine seed...

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Abstract

The invention belongs to the technical field of citrus virus-free seedling breeding, and in particular relates to a method and application of compound microorganism treatment and control of phloem bacillus pathogen in seedlings. Citrus huanglongbing is a devastating disease in citrus production all over the world, and its pathogen is caused by the hard-to-culture phloem bacteria. This technology involves reducing the concentration of pathogens or controlling the pathogens of phloem bacilli in citrus seedlings, so as to provide scions for cultivating virus-free seedlings or provide non-toxic or low-toxic shoot tip materials for grafting and detoxification of shoot tip microbuds, which can greatly improve detoxification Efficiency, providing an effective means to solve the bottleneck problem of citrus virus-free breeding. Through the high-intensity application of compound bacillus, this application seeks a method to control and reduce the pathogenic concentration of Huanglongbing, laying an important foundation for the follow-up microbud grafting and detoxification work, and achieving the goal of reducing the pathogenic concentration of the female plant and improving the detoxification effect The purpose is to quickly provide safe and healthy stem-tip scions for the industry, and then realize the purpose of disease-free improved citrus varieties.

Description

technical field [0001] The invention belongs to the technical field of citrus virus-free seedling breeding, and in particular relates to a method and application of compound microorganism treatment and control of phloem bacillus pathogen in seedlings. Background technique [0002] Citrus Huanglongbing (HLB) has seriously hindered the development of the global citrus industry. Due to the difficulty in culturing its pathogenic bacteria in vitro, the research on the pathogenic mechanism of HLB has slowed down. At present, there is no targeted control method. [0003] Citrus Huanglongbing (HLB), also known as citrus huanglongbing (HLB), is a plant disease caused by bacteria of the genus Candidatus Liberibacter. At present, citrus huanglongbing has occurred and spread in more than 50 countries in Asia, Africa, Oceania, South America, and North America, and it is still expanding. In the United States and Brazil, Huanglongbing has had a devastating impact on the development of the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01G13/00A01G2/30A01G17/00C12N1/20C12R1/07
CPCA01G13/00A01G17/005A01G2/30C12N1/20C12N1/205C12R2001/07
Inventor 姜玲徐媛媛胡党振南京
Owner HUAZHONG AGRI UNIV
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