Rhizobium v9-2 and its application
A rhizobia, a typical technology, applied in the field of microorganisms, can solve problems such as not necessarily good results, and achieve the effect of strong nitrogen fixation ability and strong stress resistance
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Embodiment 1
[0021] The isolation, purification and preservation of embodiment 1 Rhizobium V9-2
[0022] From the wild purple vetch collected in Shuangshi Village, Meiyu Town, Yanyuan County, Liangshan Prefecture, Sichuan Province, select the large and full red nodules on the main roots of robust plants, wash the nodules, and pick off some root barks, and carry out surface disinfection: use ethanol ( 95%) soaked for 5 minutes, poured out, then soaked in mercuric chloride (0.1%) for 5 minutes, and finally rinsed 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...
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. Regularly replenish sterilized nitrogen-free nutrient solution, inoculate Rhizobium V9-2 into a sand culture device, use a 300mL plastic cup for the sand culture device, use vermiculite as the substrate, and use the same variety of plants that were not inoculated with Rhizobium V9-2 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 V9-2.
[0027] (1) Bacteria culture: Inoculate Rhizobium bacterium V9-2 in YMA liquid medium, pl...
Embodiment 3
[0037] The stress resistance of embodiment 3 rhizobia V9-2
[0038] The stress resistance of rhizobia V9-2 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 Rhizobium V9-2 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. The plates for acid and alkali resistance test, salt resistance test, and suitable growth temperature range determination were all 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 formula for salt t...
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