Method for promoting the growth of clover and/or increasing the yield of clover and inoculum used therefor
A technology of clover and yield, applied in the field of agricultural microorganisms, can solve the problems of soil nitrogen consumption and soil fertility decline, and achieve the effects of increasing plant fresh weight and dry weight, increasing hay yield, and increasing fresh grass yield
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Embodiment 1
[0038] Example 1, Isolation and Identification of Rhizobium trifolii (Rhizobium trifolii) 92-29CGMCC No.15533
[0039] 1. Isolation of strains
[0040]Collect wild white clover root nodules from Qujing area, Yunnan Province, use a flat plate (get mannitol 5g, glycerol 5g, yeast extract powder 1g, dipotassium hydrogen phosphate 0.5g, calcium sulfate 0.2g, magnesium sulfate 0.2g, sodium chloride 0.1g , the mass content is 1% ammonium molybdate aqueous solution 1ml and the mass content is 1% boric acid aqueous solution 1ml, 0.5% Congo red 1ml, 20g agar, distilled water 1L, pH value is 6.8-7.0) streak separation obtains bacterial strain 92-29 .
[0041] 2. Identification of strains
[0042] 2.1. Morphological identification
[0043] The strain 92-29, which is in the logarithmic growth phase and the colony size is stable, is isolated and purified in the above step 1, and the single colony state is observed, mainly including the size, color, transparency, humidity, surface state ...
Embodiment 2
[0050] Embodiment 2, the preparation of Rhizobium clover 92-29 inoculum
[0051] 1. Incline culture of Rhizobium clover 92-29
[0052] The Rhizobium cloverii 92-29 of Example 1 was picked and inoculated on a solid medium for slant culture, and cultured at 28° C. for 56 hours to obtain the Rhizobia cloverii 92-29 cultured on a slant.
[0053] 2. Activation of bacteria
[0054] Pick the Rhizobia cloverii 92-29 cultured on the slant in step 1, inoculate it in 200 mL of liquid medium, and culture it with shaking at 28° C. and 150 rpm for 48 hours to obtain the Rhizobia cloverii 92-29 bacterium liquid.
[0055] 3. Seed tank cultivation
[0056] Get 3L of the Rhizobium clover 92-29 bacterium liquid of step 2, insert it into a 100L seed tank (containing 60L liquid medium) and carry out seed culture, and cultivate it with aeration and stirring at 28°C and 150rpm for 48 hours to obtain the Rhizobium clover 92 -29 seed liquid.
[0057] 4. Fermentation tank culture
[0058] Accordin...
Embodiment 3
[0062] Embodiment 3, the growth situation comparison (tolerance test) of Rhizobium clover 92-29 at different pHs on the acid medium
[0063] After the above liquid medium is sterilized, use sterile 1% NaOH solution and 1% HCl solution to adjust the pH of the medium to 4.2, 4.4, 4.5, 4.6, 4.8, 5.0, 5.5, 6.0, 7.0 to obtain different pH values The liquid medium of each treatment was repeated three times. The strains of Rhizobium cloverii 92-29 and R. trifolii ACCC 18019 were respectively inoculated in liquid medium with different pH values. After 48 hours of shaking culture at 150rpm and 28°C, , using the liquid culture medium without inoculation of each treatment as a blank control to determine its OD 600 value. The results are shown in Table 1, showing that Rhizobium clover 92-29 can grow in the range of pH 4.2-7.0, Rhizobium clover ACCC 18019 can only grow in the range of pH 4.8-7.0, and can grow in the pH range of 4.8-7.0. In the range of 4.2-7.0, the growth status of Rhizo...
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