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Trichoderma afroharzianum Ta97 and application thereof in straw returning

A ta97 and straw technology, which is applied to Trichoderma harzianum Ta97 in Africa and its application field in straw returning, can solve the problems of high research cost of compound bacterial strains, difficult and unsatisfactory application, and reduce manual handling of straw. , the effect of saving labor and cost, avoiding site restrictions

Active Publication Date: 2021-02-05
BIOTECH CENT OF SHANDONG ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among the resource utilization methods of straw, animal feed and compost are commonly used. Among them, compost is the main resource utilization method. However, compost requires a special composting site, which limits straw treatment. Therefore, seeking A more reasonable way of resource utilization is of great significance to straw treatment
[0003] Straw returning to the field is a cost-effective method of straw treatment. However, the lignin in the straw combines with cellulose and hemicellulose through ester bonds, and interweaves to form a strong tissue to resist adversity, resulting in a long degradation cycle after the straw is returned to the field. , not only cannot effectively release nutrients to the soil, but the undegraded straw will also hinder the development of the next crop at the seedling stage
[0004] In the prior art, when the straw is returned to the field, microorganisms containing lignocellulose-degrading enzymes are often used to accelerate the degradation of the returned straw. Therefore, multiple strains of bacteria are usually combined to form a composite strain. However, due to the different growth and enzyme production conditions of each strain, the fermentation conditions of the composite strain are harsh. In addition, the research cost of the complex strain is high, thus It is difficult to realize the application of compound bacteria system in straw returning to field, which is also the main factor restricting straw returning to field

Method used

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  • Trichoderma afroharzianum Ta97 and application thereof in straw returning
  • Trichoderma afroharzianum Ta97 and application thereof in straw returning
  • Trichoderma afroharzianum Ta97 and application thereof in straw returning

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Embodiment 1

[0029] The isolation and identification of embodiment 1 bacterial strain

[0030] (1) Source of strain

[0031]The Trichoderma harzianum Ta97 of the present invention is isolated and screened from the farmland soil of the corn-wheat rotation, and the sampling depth is 20 cm.

[0032] The collected soil samples were diluted with sterile water containing 0.3% Tween-80 to 10 -1 ~10 -7 times, 100 μL was taken from each gradient dilution to coat the rose bengal sodium agar medium plate, and cultured at a constant temperature of 25°C for 3 days. A single colony was picked from each serially diluted plate, and the species was identified by morphological microscope observation and translation elongation factor 1-alpha gene (tef1) comparison, and the strain numbered Ta97 was isolated The dominant strain was identified as Trichoderma afroharzianum.

[0033] For the colony status of Ta97, see figure 1 , in the figure, A is the front view of the colony, B is the reverse view of the c...

Embodiment 2

[0036] Example 2 strain screening

[0037] (1) Trichoderma harzianum Ta97 obtained in Example 1 was cultivated at a constant temperature of 25° C. for 3 days on a PDA plate;

[0038] (2) screening

[0039] Using plate screening, the process is as follows:

[0040] ① Screen peroxidase-producing strains on an aniline blue plate: use a sterile puncher to take the cultured colonies on the PDA plate, inoculate them on the aniline blue plate, prepare 3 replicate plates, and culture them upside down at 25±1°C , Record whether there is decolorization and transparency on the aniline blue plate, the decolorization is recorded as +, otherwise it is recorded as -. According to the diameter of the transparent area, select the strain with strong peroxidase-producing ability, and record the time.

[0041] Aniline blue solid medium: RB brilliant blue was filtered separately to make 2% mother solution (200 times), and added to PDA at 0.01% (V / V) under sterile conditions.

[0042] After cul...

Embodiment 3

[0053] The enzyme activity assay of embodiment 3 bacterial strain metabolites

[0054] ① Detection of lignin peroxidase activity: Activate Trichoderma harzianum Ta97 on a PDA plate, pick Trichoderma harzianum Ta97 on the plate to prepare 1×10 7 The conidia suspension of each / mL was inoculated in the veratrol culture solution by 1% (V / V) (the culture solution included: starch 0.6g, yeast powder 7.5g, wheat straw powder 0.5g, KH 2 PO 4 4g, MgSO 4 ·7H 2 O 0.244g, CaCl 2 0.066g, veratrol 2mmol, vitamin B1 2mg, trace element solution 40mL, 0.15% Tween-80, distilled water 1000mL), in 25 ± 1 ℃, 160rpm condition, cultivate 5 days, obtain fermented liquid; Centrifuge at 4°C and 12000rpm for 20min, and the supernatant is the crude enzyme solution.

[0055] Mix 600 μL of crude enzyme solution to be tested, 3.2 mL of 250 mmol / L tartrate buffer solution and 0.1 mL of 10 mmol / L veratrol solution to form a reaction solution, preheat in a water bath at 30 °C, and then add 10 mmol / L H 2...

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Abstract

The invention provides Trichoderma afroharzianum Ta97 and application thereof in straw returning. The strain is classified and named as Trichoderma afroharzianum in Latin, the strain is preserved in China General Microbiological Culture Collection Center on July 14th, 2020, the preservation unit is called CGMCC for short, the address is Courtyard 1, Beichen West Road, Chaoyang District, Beijing City, and the preservation number is CGMCC No.19930. The invention further discloses the application of the Trichoderma afroharzianum Ta97 in the straw returning. The Trichoderma afroharzianum Ta97 is applied to the straw returning, in a laboratory test, the average weight loss rate of straw reaches 80.40% within 30 days, and in a field test, the straw can be fully degraded within 30 days to reach the fourth level of decomposition, nutrition is provided for growth of next-stubble crops, and the purposes of straw returning cyclic utilization and fertilizer cost saving are achieved.

Description

technical field [0001] The invention relates to the technical field of microorganism application, in particular to a African Trichoderma harzianum Ta97 and its application in straw returning to field. Background technique [0002] Crop straw is one of the main solid wastes in agricultural production, with an annual output of about 800 million tons in my country. Straw contains a large amount of organic substances such as fiber, protein and fat, and has resource utilization value. Among the resource utilization methods of straw, animal feed and compost are commonly used. Among them, compost is the main resource utilization method. However, compost requires a special composting site, which limits straw treatment. Therefore, seeking A more reasonable way of resource utilization is of great significance to straw treatment. [0003] Straw returning to the field is a cost-effective method of straw treatment. However, the lignin in the straw combines with cellulose and hemicellul...

Claims

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Application Information

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IPC IPC(8): C12N1/14C05F17/20C12R1/885
CPCC12N1/14C05F17/20C12R2001/885C12N1/145Y02W30/40
Inventor 吴晓青张新建周方园赵晓燕张广志范素素王加宁
Owner BIOTECH CENT OF SHANDONG ACAD OF SCI
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