CN112322501A - Trichoderma africanum Ta97 and application thereof in straw returning - Google Patents
Trichoderma africanum Ta97 and application thereof in straw returning Download PDFInfo
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Abstract
The invention provides a Trichoderma africanum Ta97 strain and application thereof in straw returning, wherein Latin classification of the strain is named as: the Trichoderma afroharizanum strain is preserved in China general microbiological culture Collection center (CGMCC) in 14 days 07-14.2020, and the preservation unit is CGMCC, and the address is as follows: xilu No.1 Hospital, Beijing, Chaoyang, North Chen, with a deposit number: CGMCC No.19930, application of Trichoderma africanum Ta97 in straw returning. The Trichoderma africanum Ta97 is applied to straw returning, in a laboratory test, the average weight loss rate of the straw reaches 80.40% within 30 days, in a field test, the straw can be fully degraded within 30 days to reach the maturity grade 4, nutrition is provided for the growth of next-stubble crops, and the purposes of straw returning recycling and fertilizer cost saving are achieved.
Description
Technical Field
The invention relates to the technical field of microorganism application, in particular to Trichoderma africanum Ta97 and application thereof in straw returning.
Background
Crop straws are one of main solid wastes in agricultural production, and the annual output of China is about 8 hundred million tons. The straws contain a large amount of organic substances such as fiber, protein, fat and the like, and have resource utilization value. In the resource utilization mode of the straws, two modes of animal feed and compost are commonly used, wherein the compost is a main resource utilization mode, however, the compost needs a special compost field, so that the straw treatment is limited, and therefore, the search for a more reasonable resource utilization mode has important significance for the straw treatment.
The return of the straws to the field is a straw treatment mode which saves the cost, however, lignin in the straws is combined with cellulose and hemicellulose through ester bonds and interweaved to form firm tissues to resist the adverse environment, so that the degradation period is too long after the straws are returned to the field on site, nutrient substances cannot be effectively released to the soil, and the straws which are not completely degraded can also block the seedling development of next-stubble crops.
In the prior art, when straw is returned to the field on the spot, the returned straw is degraded at an accelerated speed by using microorganisms containing lignocellulose degrading enzyme systems, but most degrading bacteria have single degrading function and cannot meet the requirement of the straw degradation of the returned straw on the spot, so that a plurality of strains are combined to form a composite bacterial system, but the fermentation conditions of the composite bacterial system are harsh due to different growth and enzyme production conditions of each strain, and in addition, the research cost of the composite bacterial system is high, so that the application of the composite bacterial system in the field on the spot of the straw is difficult to realize really, and the main factor for limiting the field on the spot of the straw is also limited.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a Trichoderma africanum Ta97 strain and application thereof in straw returning, wherein the Trichoderma africanum Ta97 is named as Latin classification: the Trichoderma afroharizanum strain is preserved in China general microbiological culture Collection center (CGMCC) in 14 days 07-14.2020, and the preservation unit is CGMCC, and the address is as follows: xilu No.1 Hospital, Beijing, Chaoyang, North Chen, with a deposit number: CGMCC No. 19930. The trichoderma africanum Ta97 has manganese peroxidase, lignin peroxidase, cellulase and hemicellulase activities simultaneously. The Trichoderma africanum Ta97 is applied to straw returning, the average weight loss rate of the straw in 30 days in a laboratory degradation test on the straw reaches 80.40%, the straw can be fully degraded in 30 days in a field test to reach the maturity level 4, and the straw can be fully degraded in the field returning; and the fertilizer which is beneficial to the absorption of crops is obtained, the nutrition is provided for the growth of the next crops, and the aim of saving the fertilizer cost is fulfilled.
The technical scheme of the invention is as follows:
a strain of Trichoderma africanum Ta97, which is classified and named as Latin: the Trichoderma afroharizanum strain is preserved in China general microbiological culture Collection center (CGMCC) in 14 days 07-14.2020, and the preservation unit is CGMCC, and the address is as follows: xilu No.1 Hospital, Beijing, Chaoyang, North Chen, with a deposit number: CGMCC No. 19930.
Further, the tef1 gene sequence of the Trichoderma africanum Ta97 is as follows:
AGAAGGTAACCTTCAACTGATTTTCGCCTCGATTCTTCCTCTCTTCACATTCAATTGTGCCCGACAATTCTGCAGAGAATTTTCGTGTCGACAATTTTTCATCACCCCGCTTTCCATTACCCCTCCTTTCCAGCGACGCAAATTTTTTTTTCTGTCGTTTGGTTTTTAGTGGGGTTCTCTGTGCAACCCCACTAGCTCCCTGCTTTTTCCTGCTTCACTCTCACTTCCTCGTCATCATTCAACGTGCTCTGCGTCTTTGGTCATTCAGCGACGCTAACCACTTTTCCATCAATAGGAAGCCGCCGAACTCGGTAAGG。
further, the trichoderma africanum Ta97 has manganese peroxidase, lignin peroxidase, carboxymethylcellulase and hemicellulase activities simultaneously; wherein, manganese peroxidase and lignin peroxidase are involved in decomposing lignin, carboxymethyl cellulose is involved in decomposing cellulose, and xylanase is involved in decomposing hemicellulose.
Furthermore, the activity of the manganese peroxidase is 3.52IU/mL, the activity of the lignin peroxidase is 2.43IU/mL, the activity of the carboxymethyl cellulose is 3.76IU/mL, and the activity of the xylanase is 61.65 IU/mL.
The application of the Trichoderma africanum Ta97 in straw returning is provided.
Further, in the application, the application of the microbial inoculum prepared by Trichoderma africanum Ta97 in straw returning is specifically provided.
Preferably, the microbial inoculum prepared by Trichoderma africanum Ta97 is wettable powder of Trichoderma africanum Ta97, and in the wettable powder, the effective viable count is more than or equal to 5 multiplied by 109cfu/g。
Further, the preparation method of the wettable powder of Trichoderma africanum Ta97 comprises the following steps:
(1) preparing a seed solution: inoculating Ta97 preserved at the temperature of-80 ℃ to a PDA solid plate, activating for 3 days at the temperature of 25 ℃, taking hyphae from the edge of a colony, inoculating the hyphae to the PDA solid plate again, culturing for 3 days at the temperature of 25-28 ℃, and repeatedly taking the hyphae for culturing once to obtain an activated strain; culturing the activated strain on a PDA (personal digital Assistant) plate at 25-28 ℃ for 10 days under 12-hour illumination and 12-hour darkness, and preparing 107Inoculating the conidium suspension in PDB culture solution at a volume ratio of 1:100, shake culturing at 25 deg.C and 180rpm for 40-60 hr to obtain seed solution with viable count of 1 × 104～1×105cfu/mL；
(2) Preparing trichoderma powder: inoculating 12-22% of seed liquid by volume percentage into a fermentation tank filled with a liquid fermentation culture medium for fermentation, wherein the fermentation temperature is 28-30 ℃ before 12 hours, the fermentation temperature is 24-27 ℃ after 12 hours, the initial pH is 3.5-5.5, the stirring speed is 200-300 r/min, the ventilation quantity is 10-15L/min, and the fermentation time is 60-80 hours; the number of viable bacteria in the fermentation product is 1 × 108～1×109cfu/mL; vacuum freeze drying the fermented liquid to obtain trichoderma powder with viable count not less than 1 × 1010cfu/g；
(3) Preparation: mixing trichoderma powder and an auxiliary agent according to the following parts by weight: 30-50 parts of trichoderma powder, 30-50 parts of diatomite, 2-4 parts of a wetting agent, 3-6 parts of a dispersing agent, 0.2-0.5 part of an adhesive and 1-2 parts of zinc sulfate; mixing the above materials, and making into tablet.
Further, in the step (2), the liquid fermentation medium comprises the following components in parts by weight: 50-100 parts of blasting corn straw powder, 50-100 parts of blasting wheat straw powder, 30-50 parts of wheat bran powder, 10-30 parts of glucose, (NH)4)2SO40.5 to 1 part of KH2PO40.02 to 0.04 part by weight of MgSO4·7H20.03-0.05 part of O and 5000-8000 parts of water.
Furthermore, in a degradation test of the straws under laboratory conditions, the average weight loss rate of the straws in 30 days reaches 80.40%, and in a field test, the straws can be fully degraded in 30 days to reach 4-grade maturity.
Compared with the prior art, the invention has the beneficial effects that:
1. the invention provides a Trichoderma africanum Ta97 strain, wherein the Trichoderma africanum Ta97 strain simultaneously has four degrading enzymes of manganese peroxidase, lignin peroxidase, carboxymethyl cellulose and xylanase, the activity of the manganese peroxidase is 3.52IU/mL, the activity of the lignin peroxidase is 2.43IU/mL, the activity of the carboxymethyl cellulose is 3.76IU/mL, and the activity of the xylanase is 61.65 IU/mL.
2. The Trichoderma africanum Ta97 provided by the invention has four enzyme activities simultaneously, wherein manganese peroxidase and lignin peroxidase participate in lignin decomposition, carboxymethyl cellulose participates in cellulose decomposition, and xylanase participates in hemicellulose decomposition; the degradation capability of the strain to the straws is improved by four enzyme activities, the average weight loss rate of the straws in 30 days reaches 80.40% in a laboratory degradation test of the straws, and the straws can be fully degraded in 30 days in a field test to reach the decomposed 4-grade; the problem that the requirement on fermentation conditions is harsh in the fermentation process of a composite bacterial system constructed by adopting multiple strains in the prior art is avoided, and straw returning is easier to realize.
3. The Trichoderma africanum Ta97 microbial inoculum provided by the invention can be directly used when straw is returned to the field on the spot, so that the field limitation of the existing compost is avoided, the field returning on the spot can also reduce the labor output of manual straw carrying and manual fertilization returning, and the invention has the advantages of labor and cost saving.
Drawings
In order to more clearly illustrate the embodiments or technical solutions in the prior art of the present invention, the drawings used in the description of the embodiments or prior art will be briefly described below, and it is obvious for those skilled in the art that other drawings can be obtained based on these drawings without creative efforts.
FIG. 1 is a colony and microscopic morphology of Trichoderma africanum Ta97 cultured in PDA plate at 25 deg.C for 7 days, wherein A is the front side of the colony, B is the back side of the colony, C is the conidiophore morphology, D is the conidiophore morphology, and E is the chlamydospore morphology.
FIG. 2 is a plan view of the enzyme activity assay of Trichoderma africanum Ta97, in which A is a plan view of aniline blue for screening peroxidase, B is a plan view of RB brilliant blue for screening peroxidase, C is a plan view of CMC for screening cellulase, and D is a plan view of xylan for screening hemicellulase.
Detailed Description
In order to make those skilled in the art better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
EXAMPLE 1 isolation and identification of the Strain
(1) The source of the strain
The African trichoderma harzianum Ta97 separating screen is selected from farmland soil for corn and wheat crop rotation, and the sampling depth is 20 centimeters.
Using the collected soil sampleSterile water gradient dilution to 10 containing 0.3% Tween-80-1～10-7mu.L of each of the gradient dilutions was plated on sodium rose agar and incubated at 25 ℃ for 3 days. A single colony was picked from each plate diluted in a gradient, and the species identified by morphological microscopic observation and alignment of the translation elongation factor gene (tef 1), wherein the strain numbered Ta97 was the dominant strain isolated and identified as Trichoderma africanum (Trichoderma africanum).
The colony state of Ta97 is shown in figure 1, wherein A is the front side of the colony, B is the back side of the colony, C is the conidiophores morphology, D is the conidiophores morphology, and E is the chlamydospore morphology.
The tef1 gene sequence of Ta97 is:
AGAAGGTAACCTTCAACTGATTTTCGCCTCGATTCTTCCTCTCTTCACATTCAATTGTGCCCGACAATTCTGCAGAGAATTTTCGTGTCGACAATTTTTCATCACCCCGCTTTCCATTACCCCTCCTTTCCAGCGACGCAAATTTTTTTTTCTGTCGTTTGGTTTTTAGTGGGGTTCTCTGTGCAACCCCACTAGCTCCCTGCTTTTTCCTGCTTCACTCTCACTTCCTCGTCATCATTCAACGTGCTCTGCGTCTTTGGTCATTCAGCGACGCTAACCACTTTTCCATCAATAGGAAGCCGCCGAACTCGGTAAGG。
example 2 Strain screening
(1) The Trichoderma africanum Ta97 obtained in example 1 was cultured on a PDA plate at a constant temperature of 25 ℃ for 3 days;
(2) screening
Plate screening was used, the procedure was as follows:
screening a peroxidase-producing strain by using an aniline blue plate: taking the colony cultured on the PDA plate by using a sterile puncher, inoculating the colony on an aniline blue plate, preparing 3 plates for repeated culture, performing inverted culture at the temperature of 25 +/-1 ℃, recording whether the phenomenon of decoloration and transparence exists on the aniline blue plate, and recording the decolored colony as plus or minus on the aniline blue plate. According to the diameter of the transparent zone, the strains with strong peroxidase-producing ability were selected and the time was recorded.
Aniline blue solid medium: RB Brilliant blue was filtered separately to make 2% mother liquor (200 times) and added to PDA at 0.01% (V/V) under sterile conditions.
After cultivation, T.africana Ta97 decolorized and cleared the dye in aniline blue medium, and the clear zone on day 4 was 4.72cm in diameter, as shown in FIG. 2A.
RB brilliant blue plate screening bacterial strains producing peroxidase: taking the colony cultured on the PDA plate by using a sterile puncher, inoculating the colony on a PDA-RB brilliant blue plate, preparing 3 plates for repeated culture, performing inverted culture at the temperature of 25 +/-1 ℃, and recording whether a yellow ring is generated on the PDA-RB brilliant blue plate or not, and recording as plus or minus if the yellow ring is generated on the PDA-RB brilliant blue plate. According to the size of the yellow circle, selecting the strain with strong capability of producing peroxidase, and recording the color development time.
PDA-RB Brilliant blue solid medium: RB Brilliant blue was filtered separately to make 2% stock solution (200 times) and mixed with PDA medium in a ratio of 625mg per ml under aseptic conditions.
After cultivation, T.africana Ta97 produced yellow rings in PDA-RB brilliant blue medium with a yellow ring diameter of 5.68cm on day 4, as shown in panel B of FIG. 2.
Thirdly, screening strains producing cellulase by using a CMC flat plate: taking the colony cultured on the PDA plate by using a sterile puncher, inoculating the colony on a CMC plate, preparing 3 plates for repeating, carrying out inverted culture for 4 days at the temperature of 25 +/-1 ℃, then covering the plate with 0.1% Congo red dye solution, standing for 30min, decoloring for 1h by using 1mol/L NaCl, and observing whether each strain generates a hydrolysis transparent ring. Selecting the strain with strong cellulase producing capability according to the diameter of the hydrolysis transparent ring.
CMC solid medium: CMC 10g, (NH)4)2SO4 4g，KH2PO4 2g，MgSO4·7H20.5g of O, 1.0g of peptone, 16g of agar, 0.5g of deoxysodium cholate, 1000ml of water and autoclaving at 121 ℃ for 20 min.
After cultivation, Trichoderma africanum Ta97 produced a hydrolytic clearing circle in CMC medium, with a clearing circle diameter of 3.10cm on day 4, see FIG. 2C.
Screening strains producing hemicellulase by using a xylan plate: taking the colony cultured on the PDA plate by using a sterile puncher, inoculating the colony on a xylan plate, preparing 3 plates, repeating, and performing inverted culture at 25 +/-1 ℃ for 4 days; and then covering the flat plate with 0.1% Congo red dye solution, standing for 10min, decoloring for 2-3 times by using 2mol/L NaCl, and observing whether each strain generates a hydrolysis transparent circle. Selecting the strain with strong capability of producing the hemicellulase according to the diameter of the hydrolysis transparent ring.
Xylan solid medium: (NH)4)2SO4 2g，MgSO4·7H2O 0.5g，KH2PO41g, NaCl 0.5g, xylan 2g, agar 20g, distilled water 1000mL, natural pH value, 121 ℃ high pressure sterilization for 20 min.
After cultivation, T.africana Ta97 produced a hydrolytic clearing circle in CMC medium with a diameter of 8.51cm on day 4, as shown in FIG. 2D.
Through plate screening, Trichoderma africanum Ta97 has an enzyme system for simultaneously degrading lignin, cellulose and hemicellulose, and four enzymes in the enzyme system have enzyme activities.
EXAMPLE 3 determination of enzymatic Activity of metabolites of Strain
Detecting the activity of lignin peroxidase: activating Trichoderma africanum Ta97 on PDA plate, picking out Trichoderma africanum Ta97 on plate to prepare 1X 107Inoculating 1% (V/V) of conidium suspension into veratryl alcohol culture solution (culture solution containing starch 0.6g, yeast powder 7.5g, wheat straw powder 0.5g, KH)2PO4 4g，MgSO4·7H2O 0.244g，CaCl20.066g, 2mmol of veratryl alcohol, vitamin B12 mg, 40mL of trace element solution, 0.15% Tween-80 and 1000mL of distilled water), and culturing for 5 days at the temperature of 25 +/-1 ℃ and the rpm of 160 to obtain fermentation liquor; the fermentation broth was centrifuged at 12000rpm for 20min at 4 ℃ to give a crude enzyme supernatant.
Mixing 600 μ L of crude enzyme solution to be detected, 3.2mL of 250mmol/L tartaric acid buffer solution and 0.1mL of 10mmol/L veratrum alcohol solution to obtain reaction solution, preheating in 30 deg.C water bath, and adding 10mmol/L H2O2The reaction was started with 0.1mL of the solution, and the absorbance at 310nm was measured quickly and again 1 time after 4 min.
Veratryl alcohol is oxidized within 1min to generate 1 mu moL veratraldehyde, which is 1 enzyme activity unit. Three replicates were set up with sterilized fermentation broth as control. The detection proves that the lignin peroxidase activity of the Trichoderma africanum Ta97 is 2.43 IU/mL.
② detecting the activity of the manganese peroxidase: ta97 strain was activated on PDA plates to prepare 1X 107Inoculating 1% (V/V) of conidium suspension into veratryl alcohol culture solution, and culturing at 25 + -1 deg.C and 160rpm for 5 days to obtain fermentation broth; the fermentation broth was centrifuged at 12000rpm for 20min at 4 ℃ to give a crude enzyme supernatant.
Mixing 400 mu L of crude enzyme solution to be detected, 3.4mL of 50mmol/L sodium lactate buffer solution and 0.1mL of 1.6mmol/L manganese sulfate solution into reaction solution, preheating in water bath at 37 ℃, and then adding 1.6mmol/L H2O2The reaction was started with 0.1mL of the solution, and the absorbance at 240nm was measured quickly, and after 4min, the absorbance was measured again 1 time.
1 mu mol/L of Mn in 1min2+Conversion to Mn3+Is 1 enzyme activity unit. Three replicates were set up with sterilized fermentation broth as control. The manganese peroxidase activity of the Trichoderma africanum Ta97 is detected to be 3.52 IU/mL.
And thirdly, detecting the activity of the carboxymethyl cellulose: activation of Trichoderma africanum Ta97 on PDA plates 1X 10 preparation7Inoculating 1% (V/V) of conidium suspension into CMC culture solution (obtained by removing agar from CMC solid culture medium), and culturing at 25 + -1 deg.C and 160rpm for 7 days to obtain fermentation broth. The fermentation broth was centrifuged at 12000rpm for 10min at 4 ℃ and the supernatant was the crude enzyme. To a 2mL Eppendorf tube was added citric acid-Na containing 1% (w/v) CMC2HPO4400 mu L of buffer solution (pH 4.5), preheating in a 50 ℃ water bath for 3min, adding 100 mu L of crude enzyme solution, preserving the temperature for 30min, adding 1mL of DNS reagent to terminate the reaction, fully shaking, carrying out boiling water bath for 5min, cooling with cold water, and measuring the absorbance of the solution at the wavelength of 540 nm. Sterilized fermentation broth was used as control. Three replicates were set up. The detection proves that the carboxymethyl cellulase activity of the Trichoderma africanum Ta97 is 3.76 IU/mL.
And fourthly, detecting the activity of xylanase: activation of Trichoderma africanum Ta97 on PDA plates 1X 10 preparation7Inoculating 1% (V/V) of conidium suspension into xylan culture medium (obtained by removing agar from xylan solid culture medium), and culturing at 25 + -1 deg.C and 160rpm for 7And (4) obtaining fermentation liquor. 0.5mL of 20-fold diluted fermentation broth is sucked and added into 1% xylan solution prepared by disodium hydrogen phosphate-lemon buffer (pH 4.8), and enzymolysis is carried out for 30min at 50 ℃. Adding 3mL of DNS reagent after enzymolysis, heating in a boiling water bath for 5min, then rapidly cooling with ice water, supplementing distilled water to 25mL, measuring absorbance at 540nm, setting three biological repetitions, and calculating according to the following formula by taking sterilized fermentation liquor as a reference:
in the formula: w is the content of xylose produced by enzymolysis, mg;
n is the dilution multiple of the fermentation liquor;
enzymolysis time 30, min;
v is the volume of the reaction solution, mL.
Through detection, the xylanase activity of Trichoderma africanum Ta97 is 61.65 IU/mL.
Example 4 preparation of Trichoderma africanum Ta97 inoculum
The preparation method of the wettable powder of Trichoderma africanum Ta97 comprises the following steps:
(1) preparing a seed solution: inoculating Ta97 preserved at-80 ℃ to a PDA solid plate, activating for 3 days at 25 ℃, taking hyphae from the edge of a colony, inoculating the hyphae to the PDA solid plate again, culturing for 3 days at 25 ℃, and repeatedly taking the hyphae for culturing once again to obtain an activated strain; culturing the activated strain on PDA plate at 25 deg.C under 12 hr illumination and 12 hr dark condition for 10 days to obtain 107Inoculating conidium suspension at volume ratio of 1:100 into PDB culture solution, shake culturing at 25 deg.C and 180rpm for 50 hr to obtain seed solution with viable count of 5 × 105cfu/mL；
(2) Preparing trichoderma powder: inoculating the seed liquid with the volume percentage of 18% into a fermentation tank filled with a liquid fermentation culture medium for fermentation, wherein the fermentation temperature is 29 ℃ before 12 hours, the fermentation temperature is 25 ℃ after 12 hours, the initial pH is 4.2, the stirring speed is 200-300 r/min, the ventilation volume is 10L/min, and the fermentation time is 70 hours; the number of viable bacteria in the fermentation product is 1 × 109cfu/mL; vacuum freeze drying the fermented liquid to obtain trichoderma powder with viable count not less than 1 × 1010cfu/g；
The liquid fermentation medium comprises the following components in parts by weight: 80 parts of blasting corn straw powder, 80 parts of blasting wheat straw powder, 40 parts of wheat bran powder, 20 parts of glucose, (NH)4)2SO40.7 part of KH2PO40.03 part of MgSO (MgSO)4·7H20.04 part of O and 7000 parts of water;
(3) preparation: mixing trichoderma powder and an auxiliary agent according to the following parts by weight: 40 parts of trichoderma powder, 40 parts of diatomite, 2.6 parts of a wetting agent, 4.8 parts of a dispersing agent, 0.3 part of an adhesive and 1.5 parts of zinc sulfate; mixing the above materials; the effective viable count is more than or equal to 5 multiplied by 109cfu/g。
Example 5 preparation of Trichoderma africanum Ta97 inoculum
(1) Preparing a seed solution: inoculating Ta97 preserved at-80 ℃ to a PDA solid plate, activating for 3 days at 25 ℃, taking hyphae from the edge of a colony, inoculating the hyphae to the PDA solid plate again, culturing for 3 days at 27 ℃, and repeatedly taking the hyphae for culturing once again to obtain an activated strain; culturing the activated strain on PDA plate at 27 deg.C under 12 hr illumination and 12 hr dark condition for 10 days to obtain 107Inoculating conidium suspension at volume ratio of 1:100 into PDB culture solution, shake culturing at 27 deg.C and 180rpm for 40 hr to obtain seed solution with viable count of 1 × 104cfu/mL；
(2) Preparing trichoderma powder: inoculating 12% of seed liquid by volume into a fermentation tank filled with a liquid fermentation culture medium for fermentation, wherein the fermentation temperature is 28 ℃ before 12 hours, the fermentation temperature is 24 ℃ after 12 hours, the initial pH is 3.5, the stirring speed is 200-300 r/min, the ventilation volume is 12L/min, and the fermentation time is 60 hours; the number of viable bacteria in the fermentation product is 1 × 108cfu/mL; vacuum freeze drying the fermented liquid to obtain trichoderma powder with viable count not less than 1 × 1010cfu/g；
The liquid fermentation medium comprises the following components in parts by weight: 50 parts of blasting corn straw powder, 50 parts of blasting wheat straw powder, 30 parts of wheat bran powder, 10 parts of glucose, (NH)4)2SO40.5 part of KH2PO40.02 part of MgSO (MgSO)4·7H20.03 part of O and 5000 parts of water;
(3) preparation: mixing trichoderma powder and an auxiliary agent according to the following parts by weight: 30 parts of trichoderma powder, 30 parts of diatomite, 2 parts of wetting agent, 3 parts of dispersing agent, 0.2 part of adhesive and 1 part of zinc sulfate; mixing the above materials; the effective viable count is more than or equal to 5 multiplied by 109cfu/g。
Example 6 preparation of Trichoderma africanum Ta97 microbial inoculum 3
(1) Preparing a seed solution: inoculating Ta97 preserved at-80 ℃ to a PDA solid plate, activating for 3 days at 25 ℃, taking hyphae from the edge of a colony, inoculating the hyphae to the PDA solid plate again, culturing for 3 days at 28 ℃, and repeatedly taking the hyphae for culturing once again to obtain an activated strain; culturing the activated strain on PDA plate at 28 deg.C under 12 hr illumination and 12 hr dark condition for 10 days to obtain 107Inoculating conidium suspension at volume ratio of 1:100 into PDB culture solution, shake culturing at 25 deg.C and 180rpm for 60 hr to obtain seed solution with viable count of 1 × 105cfu/mL；
(2) Preparing trichoderma powder: inoculating the seed liquid with the volume percentage of 22% into a fermentation tank filled with a liquid fermentation culture medium for fermentation, wherein the fermentation temperature is 30 ℃ before 12 hours, the fermentation temperature is 27 ℃ after 12 hours, the initial pH is 5.5, the stirring speed is 200-300 r/min, the ventilation volume is 15L/min, and the fermentation time is 80 hours; the number of viable bacteria in the fermentation product is 5 × 109cfu/mL; vacuum freeze drying the fermented liquid to obtain trichoderma powder with viable count not less than 1 × 1010cfu/g；
The liquid fermentation medium comprises the following components in parts by weight: 100 parts of blasting corn straw powder, 100 parts of blasting wheat straw powder, 50 parts of wheat bran powder, 30 parts of glucose, (NH)4)2SO41 part of KH2PO40.04 part of MgSO 24·7H20.05 part of O and 8000 parts of water;
(3) preparation: mixing trichoderma powder and an auxiliary agent according to the following parts by weight: 50 parts of trichoderma powder, 50 parts of diatomite, 4 parts of wetting agent, 6 parts of dispersing agent and 0.5 part of adhesive2 parts of zinc sulfate; mixing the above materials; the effective viable count is more than or equal to 5 multiplied by 109cfu/g。
Laboratory test under laboratory conditions, the weight loss rate of Trichoderma africanum Ta97 microbial inoculum to corn stalks is measured
Raw materials: the corn stalk, the corn variety is Zhengdan 958, collected from the colored stone farm of Jinan city, Shandong province; the microbial inoculum is the microbial inoculum provided in example 4.
And (3) experimental setting:
1. raw material preparation
(1) Blasting and drying the corn straws for later use;
(2) group setting:
treatment group: 5 treatments, named T1, T2, T3, T4 and T5; the treatment process is that the corn straws are mixed with a Trichoderma africanum Ta97 microbial inoculum; the dosage of the Trichoderma africanum Ta97 microbial inoculum is 1010cfu/kg straw;
comparison group: 2 treatments, named D1 and D2; the treatment process comprises the steps of mixing the corn straws with a commercially available straw-decomposing inoculant (the commercially available straw-decomposing inoculant mainly comprises bacillus subtilis, bacillus licheniformis, saccharomycetes, mould and metabolites thereof); the dosage of the commercial microbial inoculum is 1010cfu/kg straw; control group: 1, only corn stalks;
2. procedure of the test
The corn straws of the treatment group, the comparison group and the comparison group are respectively degraded (the degradation degree is represented by weight loss ratio) according to the following method, nylon mesh bags with the aperture of 20 meshes are respectively filled into the mixture of the treatment group (5), the comparison group (2) and the comparison group (1) for 100g respectively, the mixture is buried in a soil layer with the depth of 5-10 cm, samples are taken on the 10 th, 20 th and 30 th days after the pre-burying, and the weight loss ratio of the corn straws in the nylon bags is measured.
The weight loss rate of the straws is determined by picking out the un-decomposed, rotten and soft straws in the nylon bag, washing the straws clean, and drying the straws in a 70 ℃ oven until the straws have constant weight, wherein the weight loss rate of the straws is (the mass of the straws before culture-the mass of the straws when the straws are cultured for n days)/the mass of the straws before culture is multiplied by 100 percent. The results are shown in Table 1 below,
TABLE 1 corn stover weight loss ratio (%) -at each period with different inoculants
The combination of table 1 shows that the average weight loss rates of the trichoderma africanum Ta97 microbial inoculum to the straws at 10 th, 20 th and 30 th days are respectively 25.75%, 55.89% and 80.40%, and the decomposition speed is higher than that of the commercially available decomposed microbial inoculum, which indicates that the trichoderma africanum Ta97 microbial inoculum has good degradation capability to the corn straws and good enzyme activity persistence.
In addition, the weight loss rate of 5 treatment groups shows that the Trichoderma africanum Ta97 microbial inoculum provided by the invention has consistent degradation performance on corn straws in the same time, which shows that the enzyme system formed in the metabolite obtained by Trichoderma africanum Ta97 is stable, and the enzyme activity of the enzyme in the metabolite is stable; further illustrates the metabolite of Trichoderma africanum Ta97 of the invention, which is suitable for wide application.
Application effect of field test Trichoderma africanum Ta97 microbial inoculum on degradation of corn straws and wheat straws in field soil
Test site and soil characteristics
The test place is near the Huangwang farmland in the high and new area of Jinwang, Jining, Shandong province, and the corn-wheat rotation farmland area with flat terrain and few artificial influence factors is selected. The soil type is the brown soil, the organic matter content is about 2 percent, the total nitrogen content is about 800mg/kg, the total phosphorus content is about 700mg/kg, the total potassium content is about 18g/kg, and the pH value is about 7.
1. Test method for returning corn straws to field in situ
Test area:
and arranging a plurality of cells, wherein the area of each cell is 15 square meters, all processing cells are randomly arranged, and the processing cells are separated into protection rows.
Group setting:
the microbial inoculum is the microbial inoculum provided in example 4.
(1) Treatment group: the corn straws are returned to the field after being blasted and are mixed with a Trichoderma africanum Ta97 microbial inoculum, and 3 treatment groups are set and named T1, T2 and T3; the dosage of the African trichoderma harzianum Ta97 microbial inoculum per mu is 2.5 kg/mu;
(2) control group: the corn stalks are blasted and then returned to the field;
the specific process of returning the treatment group to the field on site is as follows:
the method comprises the steps of blasting the straws harvested with corn on site, uniformly spreading the blasted straws on the field surface, uniformly spraying an aqueous solution containing a microbial inoculum (the weight ratio of a trichoderma harzianum Ta97 microbial inoculum to water is 0.8:100) on the surfaces of the straws, simultaneously uniformly applying a proper amount of urea to ensure that the carbon-nitrogen ratio of soil is 30:1, pressing and sticking the blasted straws and soil, watering until the straws fully absorb water, and decomposing in situ; before sowing, returning the straws to the field by rotary tillage, and conventionally sowing the wheat of the next crop and managing the field.
The difference between the returning to the field of the control group and the returning to the field of the treatment group is that the Trichoderma africanum Ta97 microbial inoculum is not added.
And (3) field investigation: the field straw rotten effect is investigated on 10 th, 20 th and 30 th days of the treatment group, indexes comprise color, smell and hand feeling, the color is divided into 4 grades of medium yellow, light yellow, brown yellow and black yellow, the smell is divided into 4 grades of musty, ammonia, wine and rotten, and the hand feeling is divided into 4 grades of hard, Microsoft, soft and rotten. The results of the investigation are shown in table 2,
TABLE 2 corn stover decomposition results for control and treatment groups
The combination of the table 1 shows that in the cell applying the trichoderma africanum Ta97 microbial inoculum, the returned corn straws reach the 4 th level in evaluation from three indexes of color, smell and hand feeling during investigation on the 30 th day, the decomposition is completed, and the treatment results of 3 treatment groups are consistent; the cells in the control group do not completely become thoroughly decomposed at day 30, which shows that the metabolites in the trichoderma africanum Ta97 microbial inoculum provided by the invention can degrade the straws in the field of the straws, the degradation time is short, the degradation degrees of 3 treated cells are consistent, and the trichoderma africanum Ta97 microbial inoculum has good stability in a field test.
2. Test method for returning wheat straws to field in situ
The wheat straw is the straw remained in the field after the wheat planted when the previous crop of corn straw is thoroughly decomposed. The microbial inoculum is the microbial inoculum provided in example 4.
The test area and the group setting are the same as those in the test method for returning the corn straws to the field on site, and the position for returning the corn straws to the field on site is the same as that for returning the corn straws to the field on site, so that the test method is used for researching the degradation capability of the Trichoderma africanum Ta97 microbial inoculum on the wheat straws when the field is continuously returned to the field on site. Because there is generally no interval between the harvest of wheat and the sowing of corn, the application method of the microbial inoculum for returning wheat straw to field in situ is different from that of corn straw.
The specific process of returning the treatment group to the field on site is as follows:
and (3) blasting the straws after wheat harvesting on the spot, uniformly spreading the blasted straws on the field surface, and conventionally ploughing and sowing corns in 2 grains per hole with one hole per 25 cm. In the early seedling stage of the corn, an aqueous solution containing 1% Ta97 wettable powder is uniformly sprayed by using conventional spraying equipment, and meanwhile, a proper amount of urea is uniformly applied to the seedlings to ensure that the carbon-nitrogen ratio of soil is 25:1 and the bacterial dose per mu is 2.6 kilograms per mu. Watering to make the water content of the soil 60%. Conventional intertillage operation and subsequent field management are conventional.
The difference between the returning to the field of the control group and the returning to the field of the treatment group is that the Trichoderma africanum Ta97 microbial inoculum is not added.
And (3) field investigation: (1) investigating field decomposition effect, wherein the investigation standard is the same as the above, investigating field straw decomposition effect in 10 th, 20 th and 30 th days, and the table is 3; (2) corn emergence rate was investigated, the number of control seedlings and seedlings for each treatment were investigated on the 10 th day (peak emergence period), and the rate of emergence was calculated, as shown in table 4, where the rate of emergence% = number of seedlings/number of sowns × 100.
TABLE 3 decomposition effect of wheat straw in control and treatment groups
TABLE 4 corn emergence rate on day 10 of control and treatment groups
| T1 | T2 | T3 | CK | |
| The rate of emergence% | 90.41 | 90.75 | 89.90 | 85.11 |
The combination of the table 3 shows that when the field is continuously returned to the field in the same position, the trichoderma africanum Ta97 microbial inoculum can still fully degrade the wheat straws, and the wheat straws returned to the field reach the 4 th level through evaluation of three indexes of color, smell and hand feeling during investigation on the 30 th day, so that the decomposition is completed;
it can be seen from the combination of table 4 that the application of the trichoderma africanum Ta97 microbial inoculum accelerates the degradation of the returned wheat straws, and improves the emergence rate of the next crop of corn. The good degradation performance of the Trichoderma africanum Ta97 microbial inoculum to the wheat straws when the Trichoderma africanum Ta97 microbial inoculum is continuously returned to the field on the spot is fully demonstrated, the degradation performance of the straws by metabolites in the microbial inoculum is further demonstrated to be free from the influence of seasonal environment and crops, and further, the strong adaptability and the strong degradation performance of enzymes in the microbial inoculum to the environment and the small influence of the degradation environment can be further shown, so that the method is suitable for wide popularization and application.
Although the present invention has been described in detail by referring to the preferred embodiments, the present invention is not limited thereto. Various equivalent modifications or substitutions can be made on the embodiments of the present invention by those skilled in the art without departing from the spirit and scope of the present invention, and these modifications or substitutions are within the scope of the present invention/any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
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Claims (10)
1. A strain of Trichoderma africanum Ta97, wherein the strain is classified and named as Latin: the Trichoderma afroharizanum strain is preserved in China general microbiological culture Collection center (CGMCC) in 14 days 07-14.2020, and the preservation unit is CGMCC, and the address is as follows: xilu No.1 Hospital, Beijing, Chaoyang, North Chen, with a deposit number: CGMCC No. 19930.
2. The trichoderma africanum Ta97 of claim 1, wherein the sequence of the tef1 gene of trichoderma africanum Ta97 is:
AGAAGGTAACCTTCAACTGATTTTCGCCTCGATTCTTCCTCTCTTCACAT
TCAATTGTGCCCGACAATTCTGCAGAGAATTTTCGTGTCGACAATTTTTC
ATCACCCCGCTTTCCATTACCCCTCCTTTCCAGCGACGCAAATTTTTTTT
TCTGTCGTTTGGTTTTTAGTGGGGTTCTCTGTGCAACCCCACTAGCTCCC
TGCTTTTTCCTGCTTCACTCTCACTTCCTCGTCATCATTCAACGTGCTCT
GCGTCTTTGGTCATTCAGCGACGCTAACCACTTTTCCATCAATAGGAAGC
CGCCGAACTCGGTAAGG。
3. the trichoderma africanum Ta97 of claim 1 or 2, wherein the trichoderma africanum Ta97 has manganese peroxidase, lignin peroxidase, carboxymethylcellulase and hemicellulase activities simultaneously.
4. The trichoderma africanum Ta97 of claim 3, wherein the manganese peroxidase activity is 3.52IU/mL, the lignin peroxidase activity is 2.43IU/mL, the carboxymethyl cellulase activity is 3.76IU/mL, and the xylanase activity is 61.65 IU/mL.
5. Use of Trichoderma africanum Ta97 as claimed in any one of claims 1 to 4 in straw returning.
6. The application of the microbial inoculum prepared by the Trichoderma africanum Ta97 in straw returning is characterized in that the application of the microbial inoculum is realized by the application of the microbial inoculum.
7. The use of claim 6, wherein the microbial inoculum is a wettable powder of Trichoderma africanum Ta97, in particular Trichoderma africanum Ta97, in which the effective viable count is not less than 5 x 109cfu/g。
8. The use according to claim 7, wherein the wettable powder of Trichoderma africanum Ta97 is prepared by the following process:
(1) preparing a seed solution: inoculating Ta97 preserved at the temperature of-80 ℃ to a PDA solid plate, activating for 3 days at the temperature of 25 ℃, taking hyphae from the edge of a colony, inoculating the hyphae to the PDA solid plate again, culturing for 3 days at the temperature of 25-28 ℃, and repeatedly taking the hyphae for culturing once to obtain an activated strain; culturing the activated strain on a PDA (personal digital Assistant) plate at 25-28 ℃ for 10 days under 12-hour illumination and 12-hour darkness, and preparing 107Inoculating the conidium suspension in PDB culture solution at a volume ratio of 1:100, shake culturing at 25 deg.C and 180rpm for 40-60 hr to obtain seed solution with viable count of 1 × 104～1×105cfu/mL；
(2) Preparing trichoderma powder: inoculating 12-22% of seed liquid by volume percentage into a fermentation tank filled with a liquid fermentation culture medium for fermentation, wherein the fermentation temperature is 28-30 ℃ before 12 hours, the fermentation temperature is 24-27 ℃ after 12 hours, the initial pH is 3.5-5.5, the stirring speed is 200-300 r/min, the ventilation quantity is 10-15L/min, and the fermentation time is 60-80 hours; the number of viable bacteria in the fermentation product is 1 × 108～1×109cfu/mL; vacuum freeze drying the fermented liquid to obtain trichoderma powder with viable count not less than 1 × 1010cfu/g；
(3) Preparation: mixing trichoderma powder and an auxiliary agent according to the following parts by weight: 30-50 parts of trichoderma powder, 30-50 parts of diatomite, 2-4 parts of a wetting agent, 3-6 parts of a dispersing agent, 0.2-0.5 part of an adhesive and 1-2 parts of zinc sulfate; mixing the above materials, and making into tablet.
9. The use of claim 8, wherein in step (2), the liquid fermentation culture is carried outThe nutrient medium comprises the following components in parts by weight: 50-100 parts of blasting corn straw powder, 50-100 parts of blasting wheat straw powder, 30-50 parts of wheat bran powder, 10-30 parts of glucose, (NH)4)2SO40.5 to 1 part of KH2PO40.02 to 0.04 part by weight of MgSO4·7H20.03-0.05 part of O and 5000-8000 parts of water.
10. The use of claim 8, wherein the straw has an average weight loss of 80.40% in a laboratory degradation test for straw within 30 days, and the straw is sufficiently degraded to maturity level 4 in a field test for 30 days.
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| CN113789268A (en) * | 2021-08-26 | 2021-12-14 | 西南交通大学 | Composite microbial inoculum for efficiently degrading straws as well as preparation method and application thereof |
| CN114410483A (en) * | 2022-01-27 | 2022-04-29 | 山东农业大学 | Trichoderma harzianum and application thereof in degradation of waste orchard branches |
| WO2022105128A1 (en) * | 2020-11-20 | 2022-05-27 | 山东省科学院生态研究所(山东省科学院中日友好生物技术研究中心) | Trichoderma afroharzianum ta97 and use thereof in straw returning |
| CN115812376A (en) * | 2022-11-24 | 2023-03-21 | 石河子大学 | Method for directly crushing cotton straws and returning the crushed cotton straws to field |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107245456A (en) * | 2017-06-26 | 2017-10-13 | 中国科学院微生物研究所 | One plant of Trichoderma harzianum and its application |
| CN110527634A (en) * | 2019-08-08 | 2019-12-03 | 伽蓝（集团）股份有限公司 | One plant of Tibet source produces trichoderma harzianum strain and its application of cellulase |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018183978A1 (en) * | 2017-03-30 | 2018-10-04 | Advanced Biological Marketing, Inc. | Alleviation of corn rootworm damage with microbial seed treatments |
| CN109182137B (en) * | 2018-09-13 | 2020-03-31 | 北京市农林科学院 | Disease-preventing growth-promoting trichoderma africanum and application thereof |
| CN110699289A (en) * | 2019-10-31 | 2020-01-17 | 潍坊科技学院 | Preparation method and application of straw degradation composite microbial inoculum |
| CN111172045B (en) * | 2020-02-19 | 2021-08-31 | 中国农业科学院农业资源与农业区划研究所 | Trichoderma africanum MU153 and application thereof |
| CN112322501B (en) * | 2020-11-20 | 2022-04-15 | 山东省科学院生态研究所(山东省科学院中日友好生物技术研究中心) | Trichoderma africanum Ta97 and application thereof in straw returning |
| CN112322502B (en) * | 2020-11-20 | 2022-04-15 | 山东省科学院生态研究所(山东省科学院中日友好生物技术研究中心) | Microbial inoculum prepared from Trichoderma africanum Ta97 and application thereof in preventing and treating continuous cropping diseases |
-
2020
- 2020-11-20 CN CN202011309961.2A patent/CN112322501B/en active Active
-
2021
- 2021-04-29 WO PCT/CN2021/090911 patent/WO2022105128A1/en active Application Filing
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107245456A (en) * | 2017-06-26 | 2017-10-13 | 中国科学院微生物研究所 | One plant of Trichoderma harzianum and its application |
| CN110527634A (en) * | 2019-08-08 | 2019-12-03 | 伽蓝（集团）股份有限公司 | One plant of Tibet source produces trichoderma harzianum strain and its application of cellulase |
Non-Patent Citations (3)
| Title |
|---|
| YUPING LYU: "Selection of reliable reference genes for gene expression studies in Trichoderma afroharzianum LTR-2 under oxalic acid stress", 《JOURNAL OF MICROBIOLOGICAL METHODS》 * |
| 张家麟: "哈茨木霉产纤维素酶菌株筛选及培养基优化", 《生物技术》 * |
| 李立波: "固态发酵中2种微生物降解玉米秸秆效果的对比研究", 《农业环境科学学报》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022105128A1 (en) * | 2020-11-20 | 2022-05-27 | 山东省科学院生态研究所(山东省科学院中日友好生物技术研究中心) | Trichoderma afroharzianum ta97 and use thereof in straw returning |
| CN112877323A (en) * | 2021-03-09 | 2021-06-01 | 厦门大学 | Method for screening high-yield cellulase filamentous fungi through self-adaptive mutagenesis |
| CN112877323B (en) * | 2021-03-09 | 2023-11-24 | 厦门大学 | Method for screening high-yield cellulase filamentous fungi by self-adaptive mutagenesis |
| CN113789268A (en) * | 2021-08-26 | 2021-12-14 | 西南交通大学 | Composite microbial inoculum for efficiently degrading straws as well as preparation method and application thereof |
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| CN117004495A (en) * | 2023-08-04 | 2023-11-07 | 山东省科学院生态研究所(山东省科学院中日友好生物技术研究中心) | Trichoderma atroviride T280, screening method and application thereof |
| CN117004495B (en) * | 2023-08-04 | 2024-06-11 | 山东省科学院生态研究所(山东省科学院中日友好生物技术研究中心) | Trichoderma atroviride T280, screening method and application thereof |
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