Rhizobium for promoting growth of vicia sativa L. and application of rhizobium
A technology of pea rhizobia and arrowhead pea is applied in the field of agricultural microorganisms, which can solve the problems of consumption of soil nitrogen and decline of soil fertility.
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Embodiment 1
[0028] Example 1, Isolation and Identification of Pea Rhizobium (Rhizobium leguminosarum) JW1401 CGMCC11876
[0029] 1. Isolation of strains
[0030] From Huzhu Tu Autonomous County and Ping'an District, Haidong City, Qinghai Province, wild arrowhead pea root nodule samples were collected, and on a flat plate (get mannitol 10g, yeast extract powder 1g, dipotassium hydrogen phosphate 0.5g, calcium sulfate 0.2g, magnesium sulfate 0.2g, 0.1g of sodium chloride, 1ml of ammonium molybdate aqueous solution with a mass content of 1% and 1ml of boric acid aqueous solution with a mass content of 1%, 1ml of 0.5% Congo red, 20g of agar, dilute to 1L with water, and the pH value is 6.8-7.0) Strain JW1401 was isolated by streaking.
[0031] 2. Identification of strains
[0032] 2.1. Morphological identification
[0033] The strain JW1401, which is in the logarithmic growth phase and has a stable colony size, and is isolated and purified in the above step 1, is described for the state of...
Embodiment 2
[0040] Embodiment 2, the preparation of pea rhizobia JW1401 bacterial agent
[0041] 1. Slope culture of pea rhizobia JW1401
[0042] Rhizobia pea JW1401 of Example 1 was picked and inoculated on a solid medium for slant culture, and cultured at 28° C. for 56 hours to obtain Rhizobia pea JW1401 cultured on a slant.
[0043] 2. Activation of bacteria
[0044] Pick the Rhizobia pea JW1401 cultured on the slant in step 1, inoculate it in 500 mL of liquid medium, and culture it at 28° C. for 48 hours to obtain the Rhizobia pea JW1401 bacterial liquid.
[0045] 3. Seed jar cultivation
[0046] Take 3L of the pea rhizobium JW1401 bacterial liquid in step 2, insert it into a 100L seed tank (containing 60L liquid medium) for seed culture, and cultivate it by shaking at 30°C and 120rpm for 56 hours to obtain the pea rhizobium JW1401 seed liquid .
[0047] 4. Fermentation tank culture
[0048] Put the pea rhizobium JW1401 seed liquid in step 3 into a 1000L fermenter (containing 600...
Embodiment 3
[0112] Embodiment 3, the hydroponic test of pea rhizobia JW1401 bacterial agent inoculation arrowhead pea
[0113] Arrowhead pea adopts the filter paper bridge test tube hydroponics method. Filter paper is cut into strips, and its width is 1.6cm respectively, and is made into M type, and the central recess is made into V-shaped aperture according to the size of seedlings, and the height of M type filter paper is the length of test tube minus 4cm. The test tube used is 2.0cm×20cm.
[0114] Put the prepared nitrogen-free nutrient solution into test tubes with filter paper supports, the height of which is located in the recess of the M-shaped filter paper, repeat 4 test tubes for each treatment, and set up 4 control tubes, after filling, use durable After the high-temperature and high-pressure plastic film is sealed, it is sterilized at 121°C for 1 hour for later use.
[0115] Select the Qinghai local arrowhead pea variety Qinghai Huijian seeds with uniform size, soak them in 9...
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