Rhizobium anhuiense V2-2 and application thereof
A rhizobia, preservation number technology, applied in the field of microorganisms, can solve the problem that the effect may not be good, and achieve the effects of obvious yield increase, strong nitrogen fixation ability, and strong resistance to stress.
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Embodiment 1
[0021] Isolation, purification and preservation of embodiment 1 rhizobia V2-2
[0022] From the wild purple sweet potato collected in Fuxing Town, Pujiang County, Chengdu City, Sichuan Province, select the large and full 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 about 5 minutes, pour it out, then soak it in mercuric chloride (0.1%) for about 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...
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 Rhizobium V2-2 into the 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 Rhizobium V2-2 as the 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 V2-2.
[0027] (1) Bacteria culture: Inoculate Rhizobium V2-2 in YMA liquid medium, place on a s...
Embodiment 3
[0037] The stress resistance of embodiment 3 rhizobia V2-2
[0038] The stress resistance of Rhizobium V2-2 was mainly determined by salt tolerance, acid and alkali tolerance. 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 V2-2 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...
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