Rhizobium v14-2 and its application
A rhizobia, CCTCCM2017611 technology, applied in the field of microorganisms, can solve problems such as the effect may not be good, and achieve the effect of strong nitrogen fixation ability and strong resistance to stress
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Embodiment 1
[0021] Isolation, purification and preservation of embodiment 1 rhizobia V14-2
[0022] From the wild purple sweet potato collected in Gongping Town, Wenjiang District, Chengdu City, Sichuan Province, select the large and plump red root nodules on the main roots of robust plants, wash the root nodules, and take part of the root bark, and carry out surface disinfection: soak in ethanol (95%) After 5 minutes, pour it out, then soak it in mercuric chloride (0.1%) for 5 minutes, and finally rinse it with sterile water for 6-8 times. Put the root nodules on the sterilized white porcelain plate in the ultra-clean workbench, clip the root nodules with a bamboo stick and smear them on the plate of YMA medium, perform streaking and purification, and culture in a 28°C incubator. YMA medium formula: KH 2 PO 4 0.25g, MgSO 4 `7H 2 O 0.2g, CaCl 2` 6H 2 O 0.1g, NaCl 0.1g, ammonium molybdate (1%) 2mL, boric acid (1%) 2mL, Congo red (1%) 2.5mL, mannitol 10g, yeast powder 0.8g, agar 18-20...
Embodiment 2
[0025] The back graft test of embodiment 2 rhizobia
[0026] The sand culture method was used for the backgrafting test of rhizobium. The variety of purple sweet potato used in the test was purple sweet potato, which was purchased from Chengdu Green Grass Garden Seed Industry Co., Ltd. Carry out in the light room (temperature control 22~24 ℃, light intensity about 2800 lux, sunshine time 14h), plant and harvest 46 days. Replenish sterilized nitrogen-free nutrient solution regularly, inoculate rhizobia V14-2 into a sand culture device, use a 300mL plastic cup for sand culture device, use vermiculite as the substrate, and use the same variety of plants that were not inoculated with rhizobia V14-2 as a control CK, each treatment was repeated three times. After harvesting, the number of nodules and the dry weight of plants were used to evaluate the inoculation effect of Rhizobium V14-2.
[0027] (1) Bacteria culture: inoculate rhizobia V14-2 in YMA liquid medium, place on a shak...
Embodiment 3
[0037] The stress resistance of embodiment 3 rhizobia V14-2
[0038] The stress resistance of Rhizobium V14-2 was mainly tested for salt tolerance, acid and alkali resistance. The YMA medium was used as the basal medium, and the YMA plate cultured at pH 7 and 28°C for 7 days was used as the positive control (CK). The above-mentioned YMA slant culture of rhizobia V14-2 was scraped and washed with sterile water to make a bacterial suspension for use. The spot inoculation method was used, and each treatment was repeated 3 times. For the tests of acid and alkali resistance and salt resistance, the plates for the determination of the suitable growth temperature range were observed and recorded after being cultured at 28°C for 4 days.
[0039] (1) Medium formula for acid and alkali resistance determination: take YMA medium as the basic medium, adjust the pH value with HCl and NaOH, so that the pH value of the medium is 4.0, 5.0, 6.0, 8.0, 9.0, 10.0, 11.0 .
[0040] (2) Medium fo...
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