Microbial mixed inoculant for promoting soybean growth and application thereof
A technology of mixing inoculants and microorganisms, applied in the field of agricultural microorganisms, can solve the problems of high production cost of chemical fertilizers, high cost of chemical fertilizers, poor quality of agricultural products, etc., and achieve the effect of enhancing the ability of crops to absorb water and fertilizer
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Embodiment 1
[0040] Embodiment 1, Azospirillum brasilense (Azospirillum brasilense), Hydrogenophilus papieri Monoclonal Culture of (Hydrogenophaga palleronii)
[0041] The obtained Azospirillum brasilense (Azospirillum brasilense) was stained by Gram staining method, and the Azospirillum brasilense (Azospirillum brasilense) was Gram-negative and had a helical rod-shaped structure by optical microscopy. Colonies observed by naked eyes are characterized by round shape, low protrusions, round and complete edges, and light red color (see figure 1 and figure 2 ).
[0042] Take the nutrient gravy medium numbered (0002) of China Microorganism Culture Collection Center, take a beaker with a specification of 1L and dissolve the medium to 1L, adjust the pH of the medium to between 6.8 and 7.2 before high-temperature sterilization, and put the prepared 200ml Pour the culture medium into a 500ml Erlenmeyer flask, seal the high-temperature-resistant sealing film on the mouth of the Erlenmeyer fl...
Embodiment 2
[0045] Embodiment 2, the preparation of microbial mixed bacterial agent of the present invention
[0046] The fermented liquid containing Azospirillum brasiliensis cultivated in Example 1 and the fermented liquid containing Hydrogenophilus pareii are mixed in a volume ratio of 13:7 to form a microbial mixed bacterial agent (1L) (see Figure 5 ), the optimal concentration of Azospirillum brasiliensis was 1.9×10 8 cfu / ml; the optimal concentration of hydrogenophilic bacteria papierausus is 6.8×10 7 cfu / ml, the total bacterial concentration is not less than 1×10 8 cfu / mL.
[0047] Pour 200ml of improved AGAR medium (formulation as shown in Table 2) or BPY medium (formulation as shown in Table 3) into a 500ml Erlenmeyer flask, seal the high-temperature resistant sealing film on the mouth of the Erlenmeyer flask, and place it in a high temperature extinguisher. The bacteria are sterilized at 121°C for 30 minutes, and the above-mentioned microbial mixture can be added after coo...
Embodiment 3
[0052] Embodiment 3, plate counts the number of colonies of Azospirillum brasiliensis and Hydrogenophilus parethi in the microbial mixed bacterial agent of the present invention quantity
[0053] Get the microbial mixed bacterial agent that 1ml embodiment 2 prepares and carry out gradient dilution to 10 5 , and then take 25ul of the diluted bacterium solution and carry out coating (NA) nutrient agar plate, place it upside down and cultivate it at a biochemical constant temperature for 48h, and obtain the results as follows (see Figure 6 ):
[0054] The white dots are Azospirillum brasilense, and the black dots are Hydrogenophaga palleronii;
[0055] count to get Figure 6 There are 14 black punctuation points, Figure 6 continued There are 20 middle and black punctuation points, and the arithmetic mean is 17;
[0056] count to get Figure 6 There are 50 middle white punctuation points, Figure 6 continued There are 45 middle white punctuation points, and the arithme...
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