Application of methylotrophic Bacillus zbl-1 in the control of cotton Verticillium wilt

A ZBL-1, methylotrophic technology, applied in the fields of application, bacteria, metabolic diseases, etc., can solve the problems of poor prevention and control technology effects, and achieve the effect of strong antibacterial and good application prospects

Active Publication Date: 2018-09-11
CHINA AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 methylotrophic Bacillus zbl-1 in the control of cotton Verticillium wilt
  • Application of methylotrophic Bacillus zbl-1 in the control of cotton Verticillium wilt
  • Application of methylotrophic Bacillus zbl-1 in the control of cotton Verticillium wilt

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Isolation, screening and identification of 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 roots in Shihezi City, Xinjiang, by the Plant Protection Institute of Xinjiang Academy of Agricultural Sciences, and prepared after shaking in sterile distilled water. Suspension, use the traditional plate culture method to isolate the bacteria in the soil layer, purify the strains by the plate streak method, use different culture temperatures, pH values, and medium as enrichment conditions, and optimize and screen out a batch of well-growing bacterial strains , from which a strain numbered ZBL-1 was selected.

[0035] Separation steps:

[0036] (1) The separation medium used: LB medium, peptone 10g, yeast extract 5g, NaCl 5g, distilled water 1L, pH natural.

[0037] (2) Separati...

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 carried out using bacterial 16S rDNA universal primers to obtain an amplified product with a length of about 1.4 kb. The amplified product was recover...

Embodiment 3

[0060] Example 3: Growth Factors 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] Methylotrophic Bacillus (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, and the culture conditions were pH7.0 and cultured at a temperature of 30°C. 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-1CGMCC No.11636 as seeds is 48 hours, the optimum growth pH is 7.0, and the time for mass production of spores is 48 hours.

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Abstract

The invention discloses the application of a methylotrophic bacillus ZBL-1 in the prevention and treatment of cotton verticillium wilt, and a strain against dahliae is obtained by separating, screening, breeding and acclimating from soil samples collected in seriously diseased cotton fields in Shihezi City Methylotrophic Bacillus (Bacillus methylotrophicus) ZBL‑1 CGMCC No.11636, which has a high inhibitory effect on Verticillium, is significantly different from common methylotrophic Bacillus strains. At the same time, the screened strain ZBL‑1 1. The control effect on cotton verticillium wilt in the indoor pot test is 73.0%; in the field experiment, the control effect is 43.1%. Bacteria agent, this bacterium has very good application prospect, has practical significance and effect.

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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Patent Type & Authority Patents(China)
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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