Trichoderma viride XJ-3 strain and method for preparing cotton stalk rotten organic fertilizer by using same
A technology of decomposing organic fertilizer and viride trichoderma, which is applied in the field of agricultural microorganism application, green wood enzyme and viride XJ-3 to prepare cotton stalk decomposing organic fertilizer, can solve the problems that have not been reported, increase soil fertility, etc., and achieve strong The effect of growth and reproduction, reducing pollution and increasing fertility
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Embodiment 1
[0036] Example 1: Isolation, screening and identification of Trichodermaviride XJ-3CGMCCNO.10989
[0037] 1. Isolation and screening of strains
[0038] (1) Separation
[0039] The Trichoderma viride used in the present invention is sampled and separated from the soil of the cotton planting field in the Aksu area of southern Xinjiang, and the bacterial strains with certain degrading ability to the cotton stalks are preliminarily screened out according to the analysis of the degradation mechanism of the strains. Use the traditional plate culture method to isolate the microorganisms in the soil layer, purify the strains by the plate streak method, and use different culture temperatures, pH values, and medium as enrichment conditions to screen out a batch of well-growing microbial strains, and select one from them. The strain whose sequence number is XJ-3.
[0040] Separation steps: According to the gradient dilution method, weigh 10g of cotton soil samples in 90mL of sterile...
Embodiment 2
[0050] Example 2: Isolation, screening and identification of Trichodermaviride XJ-3CGMCCNO.10989
[0051] 1. PCR amplification of the rDNAITS segment of Trichoderma viride and its sequencing
[0052] Pick a small amount of single colonies of the XJ-3 strain, put them into an EP tube filled with 25 μL of sterile water, boil at 100°C for 8-10 minutes, and then quickly put them into the ice-water mixture for 5 minutes. Centrifuge at 10000r / min for 5min, store at 4°C, and take the supernatant when used.
[0053] Determination of rDNAITS segment gene sequence and construction of phylogenetic tree: extract the DNA of the bacterial strain according to conventional methods, dilute the universal primer with deionized water, and perform PCR amplification. The primer design is as follows:
[0054] ITS1(F): 5'-TCCGTAGGTGAACCTGCGG-3'
[0055] ITS4(R): 5'-TCCTCCGCTTATTGATATGC-3'
[0056] Determination of gene sequence of rDNAITS segment and construction of phylogenetic tree: Extract the ...
Embodiment 3
[0059] Embodiment three: Utilize Trichodermaviride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare cotton stalk decomposed organic fertilizer
[0060] Utilize Trichodermaviride (Trichodermaviride) XJ-3CGMCCNO.10989 to prepare the concrete preparation method steps of cotton stalk decomposed organic fertilizer as follows:
[0061] (1) Strain activation culture: prepare the potato dextrose agar medium of Trichodermaviride (Trichodermaviride) XJ-3CGMCCNO.10989 strain, carry out strict aseptic operation after sterilization, transfer from the inclined plane to the plate, and cultivate at 28°C for 5 sky.
[0062] (2) Primary culture: Pick a single colony from the potato dextrose agar solid medium in step (1) and transfer it to a 50mL Erlenmeyer flask containing potato dextrose liquid medium, aseptically operate at 28°C, 120r / min culture 24h-48h.
[0063] (3) Secondary expansion culture: Inoculate the above-mentioned primary culture strain into a 500mL Erlenmeyer flask filled with...
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