Bacterial strain and application thereof in prevention and treatment of fusarium wilt
A technology of Fusarium wilt bacteria and strains, applied in the field of microorganisms, can solve the problems of economic loss, the inability to achieve the ideal control effect of diseases, and the lack of breakthrough progress in disease resistance breeding research, and achieve the effect of significant antagonistic effect.
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Embodiment 1
[0043] The isolation and screening of embodiment 1 bacterial strain of the present invention
[0044] Canna indica collected from Chenggong District, Kunming City, Yunnan Province was used as a specimen, and the endophytic bacteria contained in it were isolated by surface disinfection grinding method and plate dilution separation method.
[0045] Rinse the recovered specimens with sterile water, blot dry, first soak in 75% alcohol for about 40 seconds, rinse thoroughly with sterile water, then disinfect with 0.1% mercury liter for 1 to 3 minutes, and rinse with sterile water 3 times, blot dry with sterile filter paper.
[0046] Move the sample into a sterile mortar, cut it into pieces with sterilized scissors, add 10mL of sterile water, fully grind the homogenate, after standing still for 20-30min, draw 1mL for gradient dilution, and use sterile water to gradiently dilute to 10 2 、10 3 and 10 4 , take 200 μL of the diluted solution and spread it on the NA plate, set 3 repli...
Embodiment 2
[0050] Example 2 strain identification
[0051] After the GY1 strain was cultured on LB medium for 48 hours, the diameter of the colony was 2-5 cm, it was off-white, and irregular protrusions were formed, the surface was rough and opaque, the edges were irregular, no special smell, and spores formed (see figure 1 ). Depend on figure 1 The characteristics of the colonies shown in the figure show that the strain GY1 is similar to Bacillus subtilis (Bacillus subtilis).
[0052] The 16S rDNA gene fragment of the GY1 strain was amplified by PCR using bacterial universal primers (16s(+): 5'-AGAGTTTGATCATGGCTCAG-3'; 16s(-): 5'-TAC GGTTAC CTT GTT ACG ACT T-3') to obtain The length of the sequence is 1416bp (see figure 2 , such as shown in SEQ ID NO.1).
[0053] Subsequently, the sequence was compared with Blast in the NCBI database, and the results showed that the homology between the GY1 strain and multiple Bacillus subtilis strains reached 99.93%.
[0054] Finally, through the...
Embodiment 3
[0057] Example 3 The present invention provides the application of bacterial strain GY1 in antagonizing Fusarium wilt
[0058] Experimental raw materials:
[0059] (1) Culture of antagonistic bacteria and Fusarium wilt bacteria
[0060] Use an inoculation loop to pick up the preserved bacterial solution of strain GY1 and streak it on the LB solid medium. After the bacterial solution is dry, culture it upside down in a constant temperature incubator at 30°C for 24 hours. A single colony was picked and placed in 800 μL of LB culture medium, and cultured with shaking at 180 rpm for 16 h.
[0061] Pick the hyphae of banana, cucumber, bitter gourd, tomato and Eggplant Fusarium wilt fungus to inoculate on the PDA flat plate, and cultivate it upside down in a 28°C incubator for 5 days.
[0062] experimental method:
[0063] (1) Determination of the inhibitory effect of strain GY1 on spore germination
[0064] The activated bacterial strain GY1 bacterium solution is added in the 2...
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