A kind of pea rhizobia strain vs5-1 and its application
A technology of arrowhead pea and rhizobia, applied in the field of microorganisms, to achieve the effects of obvious increase in yield, strong nitrogen fixation ability, and good affinity
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Embodiment 1
[0015] Example 1 Isolation, purification and preservation of Pea rhizobia VS5-1
[0016] From the wild arrowhead peas collected in Xingongqiao Village, Zhongjiang County, Deyang City, Sichuan Province, select the large and full red root nodules on the main roots of robust plants, wash the root nodules, and pick off part of the root bark, and carry out surface disinfection: soak in 95% ethanol for 5 minutes Eliminate surface tension, then disinfect the surface with mercury chloride (0.1% m / v) for 5 minutes, and finally rinse with sterile water for 6-8 times. Put the root nodules on a sterilized white porcelain plate under a sterile environment, 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 O0.2g, 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, yeas...
Embodiment 2
[0019] The back graft test of embodiment 2 rhizobia
[0020] The backgrafting test of rhizobia uses the sand culture method, and the arrowhead pea variety used in the test is North Sichuan arrowhead pea, which comes from Xiangshui Village, Yanjiang Town, Yanjiang District, Ziyang City, Sichuan. Carry out in the light room (temperature control 22~24 ℃, light intensity about 2800 lux, sunshine time 14h), plant and harvest 46 days. Supplement the sterilized nitrogen-free nutrient solution regularly, inoculate the rhizobia VS5-1 into the sand culture device, use a 300mL plastic cup for the sand culture device, and use vermiculite as the substrate, so as not to inoculate the rhizobia VS5-1 The plants of the same variety as the control CK, each treatment was repeated three times. After harvesting, the nodule number and dry weight of the plants were used to evaluate the inoculation effect of Rhizobia VS5-1.
[0021] (1) Bacteria culture: inoculate the Rhizobium spp. VS5-1 in YMA li...
Embodiment 3
[0031] Example 3 The Stress Resistance of Pea Arrowhead Rhizobium VS5-1
[0032] The stress resistance of Rhizobium spp. VS5-1 was mainly tested for 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). Scrape and wash the above-mentioned YMA slant culture of Rhizobium spp. VS5-1 with sterile water to make a bacterial suspension for use. The spot inoculation method was used, and each treatment was repeated 3 times. Plates for acid and alkali resistance and salt resistance tests were all cultured at 28°C for 3 days, and the results were observed and recorded.
[0033] (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 .
[0034] (2) Medium formula for salt tolerance determination: take t...
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