Trichoderma strain NAU-TL11-7 and application thereof in oat growth and seed yield increase

Abstract

The invention discloses a trichoderma strain NAU-TL11-7 and an application of the trichoderma strain NAU-TL11-7 in oat growth and seed yield increase. According to the invention, a Trichoderma strain with a remarkable growth promoting effect is screened and separated, the strain NAU-TL11-7 is classified and named as Trichoderma lacumbens, and is preserved in the China Center for Type Culture Collection on November 15, 2024, the preservation address is Wuhan University, Wuhan, China, and the strain preservation number is CCTCC NO: M20242565. The trichoderma fertilizer product prepared by adopting the strain can obviously promote the growth of oat plants, improve the yield of oat seeds, reduce the application of chemical fertilizers in oat fields, realize the reduction and synergism of chemical fertilizers and the safe and efficient production of oats, and is an effective way for realizing the green and high-quality cultivation of artificial oat fields.

Description

Technical Field

[0001] The present invention belongs to the technical field of intensive livestock production, and particularly relates to a Trichoderma strain NAU-TL11-7 and its application. The Trichoderma fertilizer product prepared with this strain is a fertilizer-reducing and efficiency-increasing bacterial fertilizer product that can promote the growth of oats and increase seed yield. Background Art

[0002] Oats (Avena sativa L.) belong to the genus Avena of the family Gramineae, and are important grain and forage dual-purpose crops in northern China. Oat forage has excellent characteristics such as high forage yield and tolerance to barrenness. At the same time, it has high nutrient content, good palatability, and is suitable for green forage or hay making. It plays a huge role in solving the shortage of forage and feed in high-altitude and high-latitude areas of China. Currently, grain and forage dual-purpose oats are important materials for artificial grasslands, and fertilization is an important means for artificial grasslands. However, in the face of the increasing demand for oat cultivation and the rapid development of high-yield artificial grassland construction, the green and efficient development of China's oat forage and seed industries is restricted by the unreasonable application of chemical fertilizers.

[0003] During the cultivation process of oats, excessive application of chemical fertilizers will lead to the loss of soil nutrients in farmland, reduced fertilizer utilization rate, and pose a great threat to the environment. Reducing the application amount of chemical fertilizers and increasing their efficiency is an effective means to control soil nutrient loss without affecting economic benefits. Therefore, researching and utilizing microbial fertilizers is an effective way to achieve the reduction of chemical fertilizer application and increase efficiency, as well as the green and high-quality cultivation of artificial oat fields.

[0004] Trichoderma is a kind of fungus widely existing in the natural environment. It has been reported that it has growth-promoting effects on various plants such as tobacco, cucumber, tomato, banana, melon, wheat, corn, and pepper. Environmentally friendly Trichoderma fertilizers are green and environmentally friendly, ecological-friendly fertilizers. They are not only beneficial to the growth of crops, promoting the improvement of their yield and quality, but also conducive to reducing soil pollution and improving soil infertility and other problems. However, most of the research on Trichoderma fertilizers focuses on farmland and cash crops, and there is less research on artificial grasslands and oats. Screening Trichoderma strains with significant growth-promoting functions and preparing bacterial fertilizer products can replace part of the chemical fertilizers when applied in the field, and increase the yield of oat forage and seeds, achieving the reduction of chemical fertilizer application and increase of efficiency. Summary of the Invention

[0005] The first object of the present invention is to provide a Trichoderma strain NAU-TL11-7 with obvious growth-promoting functions and its application.

[0006] The second object of the present invention is to provide a Trichoderma fertilizer product that promotes the growth of oats and increases seed yield, reduces the application amount of chemical fertilizers and increases efficiency.

[0007] The third object of the present invention is to provide a method for reducing the amount of chemical fertilizer and increasing efficiency to promote the growth of oats and increase seed yield.

[0008] The object of the present invention can be achieved by the following technical solutions:

[0009] In the first aspect, the present invention claims protection for a strain of Trichoderma NAU-TL11-7, which is classified and named as Trichoderma lacuwombatense. It was deposited at the China Center for Type Culture Collection on November 15, 2024. The deposit address is: Wuhan University, Wuhan, China, and the strain deposit number is: CCTCC NO: M20242565.

[0010] The NAU-TL11-7 strain was identified as Trichoderma lacuwombatense by morphological, physiological and biochemical characteristics and fungal ITS. Its optimal growth conditions are: culture temperature 28°C, pH 5.0 - 6.0, carbon source is glucose or sucrose, and it is cultured on PDA medium. The Trichoderma strain NAU-TL11-7 grows rapidly. After being cultured for 55h under optimal conditions, the colony can completely cover the petri dish (9cm); the initial colony is slender and cottony, and the color is yellow; when the conidia mature, the colony gradually turns green from the center to the edge; when the mycelium slowly ages, the surface of the medium is densely covered with dark green spores.

[0011] In the second aspect, the present invention claims protection for the application of the above-mentioned Trichoderma strain NAU-TL11-7 in promoting the growth of oats and increasing seed yield.

[0012] In the third aspect, the present invention claims protection for the application of the above-mentioned Trichoderma strain NAU-TL11-7 in the preparation of Trichoderma fertilizer products. Further, the Trichoderma fertilizer product is a Trichoderma fertilizer product that can promote the growth of oats and increase seed yield while reducing the amount of chemical fertilizer and increasing efficiency.

[0013] In the fourth aspect, the present invention claims protection for a special Trichoderma fertilizer product for reducing the amount of chemical fertilizer and increasing efficiency. This fertilizer product contains the solid fermentation product of the above-mentioned Trichoderma strain NAU-TL11-7. The solid fermentation product is a solid fermentation product prepared based on solid shallow pan fermentation technology.

[0014] Further, the content of Trichoderma strain NAU-TL11-7 in the fertilizer product ≥ 4.0×10 9 CFU / g.

[0015] Further, the above-mentioned Trichoderma fertilizer product is prepared by a method including the following steps:

[0016] (1) Prepare the seed liquid of the above Trichoderma strain NAU-TL11-7, and the viable bacteria count of the seed liquid ≥ 2.0×10 8 CFU / mL;

[0017] (2) Inoculate 10% (v / m, ml / 100g) of the seed liquid prepared in step (1) into the crushed and sterilized pure corn straw, and adjust the water content and pH value until the spore production of Trichoderma strain NAU-TL11-7 in the fermentation culture system ≥ 4.0×10 9 spores / g to obtain the Trichoderma solid fermented product;

[0018] (3) Evaporate the water content of the Trichoderma solid fermented product prepared in step (2) to less than 30% under normal temperature conditions, and mechanically extrude and granulate to obtain the Trichoderma fertilizer product. The Trichoderma fertilizer product has a significant growth-promoting effect, and the matrix after sterilization of the Trichoderma fertilizer product has no growth-promoting effect.

[0019] Further, the process of preparing the seed liquid of Trichoderma strain NAU-TL11-7 in step (1) is: inoculate the Trichoderma strain NAU-TL11-7 into the PDB medium, and ferment and culture at 28 - 30 °C and 170 - 200 r / min for 4 - 7 d.

[0020] Further, in step (2), adjust the water content to 60% - 65%, the pH value to 4 - 8, and culture in a shallow tray at 28 °C for 4 - 7 d.

[0021] In the fifth aspect, the present invention claims a method for reducing chemical fertilizer application and increasing efficiency to promote the growth of oats and increase seed yield, applying the above Trichoderma fertilizer product to the oat field to promote the growth of oat plants, improve the oat seed yield, and at the same time reduce the application of chemical fertilizers in the oat field.

[0022] In the specific implementation manner of the present invention, the field test process of the above microbial fertilizer product includes the following steps:

[0023] (1) Fertilization and ploughing: The field test was carried out in Shenggaiying Village, Xincheng District, Hohhot City, Inner Mongolia Autonomous Region (location information: longitude 111.781829, latitude: 40.89902). The tested soil has an organic matter content of 9.39 g / kg, total phosphorus of 1.95 g / kg, total potassium of 11.87 g / kg, available nitrogen of 26.02 mg / kg, available phosphorus of 7.64 mg / kg, available potassium of 44.4 mg / kg, and a pH value of 8.79. The test method is a plot test, with a total of 4 treatments, 3 replicates for each treatment, a total of 12 plots. The blocks are set along the direction of soil fertility variation to ensure that the soil fertility in each block is uniform and randomly distributed. The area of the test plot is 30 m 2 .

[0024] For treatment CK1, no chemical fertilizer was applied. For treatment CK2, chemical fertilizer was applied according to local custom at a rate of 15 kg / mu. For treatment CK3 (15% substrate + 85% chemical fertilizer), the application rate of chemical fertilizer was reduced by 15%, and the chemical fertilizer and substrate were mixed and broadcast at a rate of 12.75 kg / mu of chemical fertilizer + 2.25 kg / mu of substrate. For treatment CL (15% Trichoderma fertilizer + 85% chemical fertilizer), Trichoderma fertilizer replaced 15% of the chemical fertilizer, and the chemical fertilizer and Trichoderma fertilizer were mixed and broadcast at a rate of 12.75 kg / mu of chemical fertilizer + 2.25 kg / mu of Trichoderma fertilizer. After the fertilizers were broadcast, the soil was plowed.

[0025] The chemical fertilizer was a commercially available conventional 18-46-0 diammonium phosphate compound fertilizer. The tested oat variety was Longyan No. 4. After sterilization, the Trichoderma fertilizer product could be used as a field control substrate.

[0026] (2) Sowing: The sowing method was broadcasting, and the seeding rate was 12 kg / mu.

[0027] (3) Index measurement: The plant height and forage yield were measured from the jointing stage to the heading stage of oats, and the seed yield was measured at maturity.

[0028] Advantages of the present invention:

[0029] (1) A Trichoderma strain NAU-TL11-7 was screened and isolated in situ from Inner Mongolia region, and the Trichoderma fertilizer product prepared therefrom had obvious growth-promoting function and competitive effect, and could effectively promote the rooting and growth of plants.

[0030] (2) Since the Trichoderma fertilizer product was based on solid shallow pan fermentation technology, the obtained Trichoderma fertilizer product had good biological safety of the strain, was easy to ferment, had low production cost, was green and pollution-free, was easy to promote and apply, was beneficial to the pollution-free production of oats, and was also beneficial to the export of oat products, with good application prospects.

[0031] (3) The Trichoderma fertilizer product of the present invention had a wide application range: it not only had obvious root-promoting and growth-promoting effects on leguminous forage alfalfa, but also could be applied to the cultivation of gramineous forage oat for both grain and forage, and had obvious promoting effects on the forage yield and seed yield of oats.

[0032] (4) The Trichoderma fertilizer product of the present invention played a role in reducing chemical fertilizer application and increasing efficiency in artificial oat fields, reducing the application rate of chemical fertilizer, lowering the agricultural planting cost, reducing chemical fertilizer pollution, increasing the forage and seed yields of oats, and increasing the income of farmers. Description of the drawings

[0033] Figure 1 Colony morphology diagram of growth-promoting Trichoderma NAU-TL11-7.

[0034] Figure 2 18S rDNA / ITS identification result diagram of Trichoderma NAU-TL11-7.

[0035] Figure 3 Promoting growth effect diagram of Trichoderma NAU-TL11-7 in greenhouse pot experiments.

[0036] Figure 4 Data analysis of dry weight and fresh weight promoted by NAU-TL11-7.

[0037] Figure 5 Analysis of field data (plant height, seed yield) of Trichoderma fertilizer on reducing chemical fertilizer and increasing efficiency in artificial oat fields.

[0038] Figure 6 Effect diagram of field experiment of Trichoderma fertilizer on reducing chemical fertilizer and increasing efficiency in artificial oat fields. Specific implementation manners

[0039] The present invention will be further described below in conjunction with embodiments. For experimental methods without specific conditions indicated in the following embodiments, they are generally carried out according to the well-known means in the art.

[0040] Embodiment 1

[0041] 1. Isolation, screening and identification of Trichoderma strains

[0042] Isolation of Trichoderma strains: The soil used was collected from Jarud Banner, Tongliao City, Inner Mongolia Autonomous Region (44°41'40"N, 120°31'24"E). The method of dilution coating was used to screen Trichoderma strains, and all strains were stored in glycerol at -80°C. The Trichoderma strains used were identified as Trichoderma lacuwombatense NAU-TL11-7 through morphological, physiological and biochemical characteristics and fungal ITS. Its optimal growth conditions are: culture temperature 28°C, pH 5.0 - 6.0, carbon source is glucose or sucrose, and it is cultured on PDA medium. The Trichoderma strain NAU-TL11-7 grows rapidly. After culturing for 55 h under the optimal conditions, the colony can completely cover the petri dish (9 cm); the initial colony is slender and cottony, and the color is yellow; when the conidia mature, the colony gradually turns green from the center to the edge; when the mycelium gradually ages, the surface of the medium is densely covered with dark green spores (such as Figure 1 ). The ITS identification result of NAU-TL11-7 is as Figure 2 . This strain was deposited in the China Center for Type Culture Collection on November 15, 2024, and the accession number of the strain is CCTCC NO: M20242565.

[0043] 2. Determination of the growth-promoting effect of NAU-TL11-7 in a greenhouse pot system

[0044] The spore suspension of Trichoderma strain NAU-TL11-7 was applied to the roots of alfalfa seedlings, with distilled water as the control. The growth-promoting effect of the Trichoderma strain was evaluated by observing the growth of alfalfa seedlings. The results showed that NAU-TL11-7 could significantly promote the growth of alfalfa. Under the treatment of NAU-TL11-7, the dry weight, fresh weight, and plant height of alfalfa increased by 242.16%, 207.14%, and 88.42% respectively compared with the control, with a significant increase (P≤0.05)( Figure 3 , 4).

[0045] 3. Production of Trichoderma NAU-TL11-7 biofertilizer product

[0046] (1) The growth-promoting strain NAU-TL11-7 was inoculated into PDB culture medium for liquid fermentation production. The fermentation conditions of the liquid fermentation were as follows: the fermentation temperature was 28°C, the rotation speed was 200 rpm / min, and the fermentation time was 4 - 5 d, ensuring that the viable count of the growth-promoting bacteria seed liquid ≥ 2.0×10 8 CFU / mL.

[0047] (2) 6.5% (v / m, ml / 100 g) amino acid solution was added to the crushed and sterilized pure corn straw (2 - 5 mm), and 10% of the prepared growth-promoting bacteria fermentation broth (v / m, ml / 100 g) was inoculated. The water content was adjusted to 60 - 65% (preferably 65%), and the pH value was adjusted to 4.5 - 6.0. It was cultured in a shallow tray at 28°C for 5 d, so that the spore production of Trichoderma strain NAU-TL11-7 in the fermentation culture system ≥ 2.0×10 10 spores / g, and the Trichoderma solid ferment was obtained.

[0048] (3) Under the condition of normal temperature (25±5°C), the water content of the corn straw mixture of the growth-promoting bacteria was evaporated to less than 30%, and it was mechanically extruded into pellets to obtain a biofertilizer product of Trichoderma that can promote the growth of oats and increase seed yield and reduce fertilizer input.

[0049] 4. Experiment on reducing fertilizer input and increasing efficiency of Trichoderma NAU-TL11-7 product on oats

[0050] The experiment was carried out from May 17, 2024 to August 9, 2024 in Shenggaiying Village, Xincheng District, Hohhot City, Inner Mongolia Autonomous Region (location information: longitude 111.781829, latitude: 40.89902).

[0051] The organic matter of the tested soil was 9.39 g / kg, total phosphorus was 1.95 g / kg, total potassium was 11.87 g / kg, available nitrogen was 26.02 mg / kg, available phosphorus was 7.64 mg / kg, available potassium was 44.4 mg / kg, and the pH value was 8.79. The tested oat variety was Longyan No. 4.

[0052] The test method was a plot test. A total of 4 treatments were set, with 3 replicates for each treatment, resulting in 12 plots in total. The blocks were set along the direction of soil fertility gradient to ensure that the soil fertility within each block was uniform and randomly distributed. The sowing method was broadcasting, and the area of each test plot was 30 m 2 . For the specific details of the fertilization treatments, please refer to Table 1 below. The chemical fertilizer was a commercially available 18-46-0 diammonium phosphate compound fertilizer; after sterilization, the Trichoderma fertilizer product was used as the substrate and applied as the field control.

[0053] Sowing: The sowing method was broadcasting, and the seeding rate was 12 kg / mu.

[0054] Index measurement: The plant height and forage yield were measured from the jointing stage to the heading stage of oats, and the seed yield was measured at maturity.

[0055] Table 1 Fertilizer application table for each treatment

[0056]

[0057] Note: The substrate refers to the material directly sterilized from the Trichoderma fertilizer.

[0058] The results of the field growth promotion test showed that the treatment of 15% Trichoderma fertilizer + 85% chemical fertilizer (CL) with the application of Trichoderma NAU-TL11-7 could effectively increase the seed yield, forage plant height and yield of oats on the basis of reducing the chemical fertilizer application by 15%. The seed yield of the 15% Trichoderma fertilizer + 85% chemical fertilizer (CL) treatment reached 150.6742 kg / mu, showing significant differences (P≤0.05) compared with the blank control (CK1), conventional fertilization (CK2) and 15% substrate + 85% chemical fertilizer (CK3) treatments ( Figure 6 ), with yield increases of 153.05%, 125.79% and 120.64% respectively. The forage yield of the 15% Trichoderma fertilizer + 85% chemical fertilizer (CL) treatment reached 486.35765 kg / mu, showing significant differences (P≤0.05) compared with the blank control (CK1), conventional fertilization (CK2) and 15% substrate + 85% chemical fertilizer (CK3) treatments ( Figure 6 ), with yield increases of 200.695%, 101.178% and 112.06% respectively. It is recommended to promote the application of this Trichoderma fertilizer product in oat cultivation.

Claims

1. A Trichoderma strain NAU-TL11-7, which is classified and named as Trichoderma lacuwombatense. It was deposited at the China Center for Type Culture Collection on November 15, 2024. The deposit address is: Wuhan University, Wuhan, China, and the accession number of the strain is: CCTCC NO: M 20242565.

2. Use of the Trichoderma strain NAU-TL11-7 according to claim 1 in promoting the growth of oats and increasing seed yield.

3. Use of the Trichoderma strain NAU-TL11-7 according to claim 1 in the preparation of a Trichoderma fertilizer product.

4. The application according to claim 3, characterized in that The Trichoderma fertilizer product is a fertilizer-reducing and efficiency-increasing Trichoderma fertilizer product that can promote the growth of oats and increase seed yield.

5. A Trichoderma fertilizer product, characterized in that, The fertilizer product contains the solid fermentation product of the Trichoderma strain NAU-TL11-7 according to claim 1.

6. The Trichoderma fertilizer product according to claim 5, wherein The content of Trichoderma strain NAU-TL11-7 in the described microbial fertilizer product ≥ 4.0×10 9 CFU / g.

7. The Trichoderma fertilizer product according to claim 5 or 6, characterized in that, It is prepared by a method including the following steps: (1) Prepare the seed liquid of the Trichoderma strain NAU-TL11-7 described in claim 1, and the viable count of the seed liquid is ≥ 2.0×10 8 CFU / mL; (2) Inoculate 10% of the seed solution prepared in step (1) into the pulverized and sterilized pure corn straw, and adjust the water content and pH value until the spore production of the Trichoderma strain NAU-TL11-7 in the fermentation culture system is ≥ 4.0×10 9 spores / g to obtain a Trichoderma solid fermented product; (3) Under normal temperature conditions, evaporate the water content of the Trichoderma solid fermentation product prepared in step (2) to less than 30%, and mechanically extrude and granulate to obtain the Trichoderma fertilizer product.

8. The Trichoderma fertilizer product according to claim 7, characterized in that In step (1), the process of preparing the seed liquid of the Trichoderma strain NAU-TL11-7 is: inoculate the Trichoderma strain NAU-TL11-7 into a PDB medium and ferment and culture at 28-30 °C and 170-200 r / min for 4-7 d.

9. The Trichoderma fertilizer product according to claim 7, wherein In step (2), adjust the water content to 60%-65% and the pH value to 4-8, and perform shallow pan culture at 28 °C for 4-7 d.

10. A method for reducing chemical fertilizer input and increasing efficiency in promoting the growth of oats and increasing seed yield, characterized in that, Applying the Trichoderma fertilizer product according to any one of claims 3-6 to an oat field can promote the growth of oat plants, increase the oat seed yield, and at the same time reduce the application of chemical fertilizers in the oat field.

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