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Application of Bacillus methylotrophicus ZBL-1 in control of cotton verticillium wilt

A ZBL-1, methyl nutritional type technology, applied in the application, bacteria, metabolic diseases and other directions, can solve the problem of poor control technology effect, and achieve the effect of strong antibacterial property and good application prospect.

Active Publication Date: 2016-03-30
CHINA AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Aiming at the relevant reports in the prior art about methylotrophic bacillus and its application in the prevention and treatment of cotton verticillium wilt in different regions, and the technical status quo of existing control technology with poor effect and many deficiencies, the present invention aims to provide Cotton verticillium wilt provides a new resistant strain with relatively high control ability and stable performance

Method used

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  • Application of Bacillus methylotrophicus ZBL-1 in control of cotton verticillium wilt
  • Application of Bacillus methylotrophicus ZBL-1 in control of cotton verticillium wilt
  • Application of Bacillus methylotrophicus ZBL-1 in control of cotton verticillium wilt

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Isolation, screening and identification of methylotrophic Bacillus (Bacillus methylotrophicus) ZBL-1CGMCCNo.11636

[0033] 1. Isolation and screening of strains

[0034] The methylotrophic bacillus (Bacillus methylotrophicus) used in the present invention is sampled from a large number of uninfected cotton root systems in Shihezi City, Xinjiang, by the Plant Protection Institute of Xinjiang Academy of Agricultural Sciences, and the bacterial suspension is prepared after shaking in sterile distilled water. Bacteria in the soil layer were isolated using the traditional plate culture method, the strains were purified by the plate streaking method, and a batch of well-growing bacterial strains were screened out through optimization using different culture temperatures, pH values, and culture media as enrichment conditions. A strain numbered ZBL-1 was selected out of them.

[0035] Separation steps:

[0036] (1) The separation medium used: LB medium, peptone 10g...

Embodiment 2

[0050] Example 2: Molecular level identification of Bacillus methylotrophicus ZBL-1CGMCCNo.11636

[0051] The bacterial DNA genome kit was used to extract the DNA of the target strain.

[0052] Bacterial 16SrDNA universal primer sequence:

[0053] 27F: AGAGTTTGATCMTGGCTCAG;

[0054] 1492R:GGTTACCTTGTTACGACTT.

[0055] The primers, markers and reaction system used in this study were purchased from Beijing Dingguo Changsheng Biological Company.

[0056] Table 2: PCR reaction system used to extract genomic DNA

[0057]

[0058] As shown in Table 2, pre-denaturation at 94°C for 5 min; entering cycle, denaturation at 94°C for 30 s, annealing at 55°C for 30 s, extension at 72°C for 1 min, 35 cycles; extension at 72°C for 10 min, and sequencing.

[0059] Using the extracted total DNA of ZBL-1 as a template, PCR amplification was performed using bacterial 16SrDNA universal primers to obtain an amplified product with a length of about 1.4kb. The amplified product was recovered a...

Embodiment 3

[0060] Example 3: Growth Factor of Methylotrophic Bacillus (Bacillus methylotrophicus) ZBL-1CGMCCNo.11636

[0061] Table 3: Effects of temperature, pH, salt, and antibiotics on the growth of strain ZBL-1

[0062]

[0063]

[0064] Bacillus methylotrophicus (Bacillus methylotrophicus) ZBL-1CGMCCNo.11636 was cultured according to the above method, the culture conditions of the bacteria were: the culture medium was LB solid medium, the culture conditions: pH7.0, cultured at 30°C for 48h .

[0065] From Table 3, the strain ZBL-1 is the most suitable growth factor.

[0066] Based on the above results, it can be concluded that the optimal culture time of Bacillus methylotrophicus ZBL-1CGMCCNo.11636 as seeds is 48h, the optimum growth pH is 7.0, and the time for mass spore production is 48h.

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Abstract

The invention discloses an application of Bacillus methylotrophicus ZBL-1 in control of cotton verticillium wilt. The Bacillus methylotrophicus ZBL-1 with CGMCC No.11636 and higher inhibition effect on verticillium dahlia is obtained through separation, screening, breeding and domestication of soil samples collected in the cotton fields with severe diseases in Shihezi city and is obviously different from common Bacillus methylotrophicus strains, and meanwhile, the control effect of the screened strain ZBL-1 on the cotton verticillium wilt in indoor pot culture experiments is 73.0%; the control effect of the screened strain ZBL-1 in field plot experiments is 43.1%, and whether in development of a new biopesticide or biological control agent, the Bacillus methylotrophicus ZBL-1 as a biological control material for the cotton verticillium wilt has very good application prospect and has realistic significance and functions.

Description

technical field [0001] The invention relates to the technical field of agricultural biological biocontrol, in particular to a methylotrophic bacillus and the technical field of its application in the prevention and treatment of cotton diseases. Background technique [0002] Cotton verticillium wilt is one of the world's important diseases. Since 1935, it was introduced into China by the introduction of the American cotton seed, and it has spread year by year. After the 1990s, the disease has spread to various cotton areas including Xinjiang. Especially in 1993, the disease was prevalent in various cotton areas in my country, with an incidence area of ​​2.67 million hm 2 , resulting in a loss of 100 million kg of lint (Jian Guiliang et al., 2003; Xu Li et al., 2012; Zhu Heqin et al., 2012). At present, the harm of Verticillium wilt has become the first major disease in cotton production since Fusarium wilt was effectively controlled in my country. It seriously affects the yi...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A61P3/00C12R1/07
CPCA01N63/00C12N1/205C12R2001/07
Inventor 刘海洋姚举王琦史应武王伟张仁福李春韩万里
Owner CHINA AGRI UNIV
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