An antagonistic bacteria z-18 for controlling cotton verticillium wilt and its application
A technology of Z-18 and Bacillus, applied in the field of endophytes, can solve the problems such as unseen cotton endophytes, and achieve the effects of good control effect, improved yield and quality, and simple cultivation conditions.
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Embodiment 1
[0022] Example 1: Isolation, screening and identification of Bacillus zhangzhouensis Z-18CGMCC No.15006
[0023] (1) Isolation and screening of strains
[0024] Several strains of bacteria were isolated from cotton tissue by gradient dilution method, and the plate confrontation method was used to judge whether they had antagonistic effect according to the presence or absence of inhibition zone, and the strains with certain antagonistic ability to the pathogen of cotton verticillium dahliat were preliminarily screened out. Further experiments were carried out on the strains with antagonistic effect, and the strains with stronger antagonistic ability were screened out for subsequent experiments.
[0025] A batch of microbial strains of endophytic bacteria were obtained through isolation, screening and cultivation in cotton tissue, and a strain numbered Z-18 was screened out, which belonged to Bacillus zhangzhouensis of the genus Microbacterium through microbiological classificat...
Embodiment 2
[0041] Example 2: Growth Factors of Bacillus zhangzhouensis Z-18 CGMCC No.15006
[0042] (1) According to the characteristics of Bacillus zhangzhouensis Z-18 CGMCC No.15006 provided by the present invention, its specific growth factors were determined, and the strain Z-18 was inoculated and cultured, and the results are shown in Table 2.
[0043] Table 2: Effects of temperature, pH, salt, antibiotics on the growth of strain Z-18
[0044]
[0045]
[0046] (2) Growth curve and culture conditions
[0047] The suitable pH value of strain Z-18 was screened and optimized, and the results are shown in Table 3:
[0048] Table 3: pH optimization results
[0049] 6.0 6.5 7.0* 7.4* 7.5 8.0 Z-18 0.522 0.794 0.864 0.928 0.612 0.001
[0050] Cultivate Bacillus zhangzhouensis Z-18 CGMCC No.15006 according to the method (1). The culture conditions of the bacteria are: the culture medium is TSA solid medium, and the culture conditions are: pH7.4, tempera...
Embodiment 3
[0052] Embodiment three: the fermentation process research of Bacillus zhangzhouensis Z-18 CGMCC No.15006
[0053] The purpose of obtaining preparations by fermentation is achieved through the optimization of carbon, nitrogen sources, pH value, etc.
[0054] (1) Determination of medium pH
[0055] According to the pH value of 6.0, 6.5, 7.0, 7.5, 8.0, ferment at 200r / min and temperature of 35°C, the optimum growth pH value is 7.0.
[0056] (2) Optimization of carbon and nitrogen sources
[0057] Seed liquid medium: glucose 2.0%, yeast extract 0.5%, corn steep liquor 7.5%, KH 2 PO 4 0.1%, K 2 HPO 4 0.1%, MgSO4 7H 2 O0.05%, continue to optimize on the basis of seed culture medium.
[0058] A. Carbon source optimization:
[0059] The carbon sources are glucose, sucrose, and corn steep liquor, and the other components are KH 2 PO 4 0.1%, K 2 HPO 4 0.1%, MgSO 4 ·7H 2 O 0.05%, pH7.0;
[0060] B. Nitrogen source optimization:
[0061] Nitrogen sources were peptone, yea...
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