A bacterial strain for preventing and controlling plant nematodes and its application
A plant line and strain technology, applied in the field of microbiology, can solve the problems that the research on plant nematode diseases has not been reported, and achieve good effects on the control of plant nematode activity, ecological environment and human health, and reduce the degree of pollution
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Embodiment 1
[0029] Embodiment 1: Isolation and identification of Marseilles strain Y149
[0030] (1) Separation and purification
[0031] Beef extract peptone agar medium: 3g beef extract, 10g peptone, 10g sodium chloride, 15g agar, 1000mL deionized water, pH7.4, sterilized at 121°C for 30min.
[0032] The present inventor collected plant rhizosphere soil samples from Changsha City, Hunan Province, and after natural air drying, weighed 10 g of the samples and dissolved them in 90 ml of sterile water to make a soil suspension. Then use 10-fold gradient dilution with sterile water, draw 10 -3 and 10 -4 100 μl of the soil suspension was evenly spread on the beef extract peptone medium plate. Put the above plate into the incubator at 30°C for 36h-48h, pick different colonies according to the shape, color and size of the colony, and purify them on the new beef extract peptone medium plate until the pure colonies are obtained and numbered for preservation .
[0033] (2) Morphological and p...
Embodiment 2
[0046] Embodiment 2: the preparation of microbial bacterial agent
[0047]Beef extract peptone broth (NB): 3g beef extract, 10g peptone, 10g sodium chloride, 1000mL deionized water, pH 7.4, sterilized at 121°C for 30min.
[0048] (1) Cultivation of slant seeds: Aseptically streak the thallus of Massilia sp. Y149 onto the slant of beef extract peptone agar medium, culture at 30°C for 36 hours to obtain slant seeds;
[0049] (2) Liquid seed culture: Pick an activated single colony from the slope and inoculate it in NB medium for culture, 200r / min, 30°C, culture time is 24h, to obtain the seed culture solution;
[0050] (3) Fermentation culture: insert the seed culture liquid into the NB culture medium according to the inoculation amount of 5%, at 30°C, the rotating speed is 200r / min, and the fermentation time is 72h to obtain a fermented liquid with OD600=4, and the number of viable bacteria is 3.5× 10 8 cfu / ml;
[0051] (4) Preparation of microbial agent: collect the ferment...
Embodiment 3
[0052] Example 3: Detection of the activity of lethal plant nematodes by Marseilles strain Y149
[0053] Potato dextrose agar medium (PDA): 200g of potatoes, 20g of glucose, 15g-20g of agar, 1000mL of water.
[0054] (1) Preparation of fermentation supernatant: Marseilles strain Y149 was fermented and cultured for 72 hours according to the method in Example 2 to obtain a fermentation system suspension with OD600=4, centrifuged at 10000r / min for 10min, and the supernatant was collected for later use.
[0055] (2) Preparation of nematode suspension: D. destructor potato used in this example was preserved by the Plant Nematode Laboratory of Hunan Agricultural University. Before culturing, inoculate Fusarium half naked on a PDA plate, and incubate at 25°C for 7-10 days. After the mycelia cover the entire plate, insert 100 μL of D. destructor suspension under aseptic conditions, and incubate at 25°C for 7-10 days. After the half-naked Fusarium hyphae disappeared, the nematodes cov...
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