Bacillus amyloliquefaciens for preventing and curing tomato bacterial wilt and application of bacillus amyloliquefaciens

A technology for dissolving starch spores and bacilli is applied in the field of crop disease prevention and smelting, and can solve problems such as unstable effect, food safety, and environmental pollution.

Active Publication Date: 2016-09-21
NINGBO ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the methods commonly used to prevent and control tomato bacterial wilt mainly include: chemical control, soil improvement, breeding of disease-resistant varieties, grafting and crop rotation, etc., but the effects are not...

Method used

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  • Bacillus amyloliquefaciens for preventing and curing tomato bacterial wilt and application of bacillus amyloliquefaciens
  • Bacillus amyloliquefaciens for preventing and curing tomato bacterial wilt and application of bacillus amyloliquefaciens
  • Bacillus amyloliquefaciens for preventing and curing tomato bacterial wilt and application of bacillus amyloliquefaciens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 The acquisition of biocontrol strain W118 for preventing and treating tomato bacterial wilt

[0024] Screening and Isolation of Bacillus amyloliquefaciens

[0025] (1) Soil samples were collected from the rhizosphere soil of healthy plants in fields with severe tomato bacterial wilt disease. The healthy plants were uprooted and shaken gently. After most of the soil was shaken, the remaining soil close to the roots was collected together with some fibrous roots. Pack it in a sealed bag and store it in a 4°C refrigerator for later use.

[0026] (2) Add 10 g of rhizosphere soil of healthy tomato plants into a conical flask filled with 90 mL of sterile water, vibrate vigorously on a shaker at 30° C. for 30 min, and let stand for 30 min;

[0027] (3) The supernatant was serially diluted and spread on the modified medium of beef extract peptone, each gradient was repeated 3 times, and after culturing at 30°C for 24 hours, the medium was filled with 1.0×10 8 The th...

Embodiment 2

[0030] Example 2 Identification of biocontrol strain W118 for preventing and treating tomato bacterial wilt

[0031] The strain W118 was inoculated on beef extract peptone modified medium, cultured at 30°C for 48 hours, the genomic DNA of the antagonistic strain was extracted with a bacterial genomic DNA extraction kit, and amplified with bacterial 16S rRNA primers. (F: 5'-AGAGTTTGATCCTGGCTCAG-3'R: 5'-TACCTTGTTACGACTT-3'). The reaction system is: Mg 2+ 2.5μl, 10×buffer 2.5μl, dNTP (10mmol·L -1 ) 2 μl, upstream and downstream primers (10mmol L -1 ) each 1 μl, template DNA 1 μl, Taq enzyme (5U·μl -1 )0.3μl, ddH 2 O 14.7 μl, the total system is 25 μl. All reagents used were purchased from Dalian Bao Biological Engineering Co., Ltd.

[0032] PCR reaction conditions: pre-denaturation at 95°C for 5min, denaturation at 95°C for 30s, annealing at 53°C for 1min, extension at 72°C for 2min, holding at 72°C for 10min, and a total of 30 cycles. The PCR product was detected by 1% (w...

Embodiment 3

[0035] Example 3 Antagonism of Bacillus amyloliquefaciens W118 to tomato bacterial wilt pathogen

[0036] Spot the purified strain W118 on beef extract peptone modified medium with a sterile toothpick, culture at 30°C for 12h, and use 1.0×10 8cfu / m of R. solanacearum pathogen liquid spray liquid, measure the diameter of the inhibition zone after 48h, repeat 3 times, observe the size of the inhibition zone, by the attached figure 1 and 2 It can be seen that Bacillus amylobacter W118 has a stronger inhibitory effect on tomato R. solanacearum.

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Abstract

The invention relates to bacillus amyloliquefaciens W118. The bacillus amyloliquefaciens W118 is collected in China General Microbiological Culture Collection Center (CGMCC) on March 10, 2016, and the collection number is CGMCC No.12200. The invention further provides application of the bacillus amyloliquefaciens W118 in prevention and cure of pathogenic bacteria of tomato bacterial wilt, cotton verticillium wilt, tomato early blight and cucumber corynespora cassiicola. Compared with the prior art, the bacillus amyloliquefaciens W118 and the application thereof have the advantages that the bacillus amyloliquefaciens W118 has a certain prevention and cure effect on the tomato bacterial wilt, and the highest prevention and cure rate can reach 59.2%; the highest prevention and cure rates for the cotton verticillium wilt, the tomato early blight and the cucumber corynespora cassiicola can reach 47.8%, 43.4% and 38.3% respectively.

Description

technical field [0001] The invention belongs to the technical field of crop disease prevention and treatment, and in particular relates to a bacillus amyloliquefaciens used for preventing and treating tomato bacterial wilt and its application. Background technique [0002] Tomato bacterial wilt is a devastating soil-borne bacterial disease caused by Ralstonia solanacearum. The output has declined seriously, seriously restricting the development of the tomato industry and the improvement of economic benefits. R. solanaceae has a complex strain and a wide range of hosts. It has tenacious vitality in high-temperature and high-humidity environments. It can infect more than 450 types of plants in more than 50 families. [0003] At present, the methods commonly used to control tomato bacterial wilt mainly include: chemical control, soil improvement, breeding of disease-resistant varieties, grafting and crop rotation, etc., but the effects are not stable. [3-4] . In addition, th...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/02A01N63/00A01P1/00A01P3/00C12R1/07
CPCA01N63/00A01N63/10C12N1/205C12R2001/07
Inventor 王丽丽袁赛飞吴凯李国安姚红燕金树权汪峰孙梅梅周金波谌江华
Owner NINGBO ACAD OF AGRI SCI
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