CN114431255B - Application of a kind of trichoderma aculeatus and plant growth promoting agent - Google Patents

Abstract

The invention relates to the technical field of microorganisms, in particular to application of trichoderma asperellum and a plant growth promoting microbial agent. Application of trichoderma asperellum in promoting plant growth; and the strain is utilized to prepare the microbial agent. The invention provides the preparation of the trichoderma asperellum-like microbial agent and the application of the trichoderma asperellum-like microbial agent in promoting plant growth.

Description

Application of a kind of Trichoderma aculeatus and plant growth-promoting bacterial agent

technical field

The invention relates to the technical field of microorganisms, in particular to the application of Trichoderma aculeatus and a plant growth-promoting bacterial agent.

Background technique

Trichoderma (Trichoderma spp.) is a filamentous fungus that widely exists in nature. It was first reported and described by Persoon at the end of the 18th century. Trichoderma belongs to the phylum Ascomycota, class Sordariomycetes, order Hypocreales, and family Hypocreaceae, and more than 250 species of Trichoderma have been reported so far. Trichoderma has important economic value and is widely used in various aspects of agriculture, especially as a biological control antagonist and plant growth-promoting bacteria, which has a very broad application prospect.

Wheat is an important source of human food and one of the main food crops after rice, which has a great impact on human life. Wheat is also the main ration crop in my country. In 2019, the sown area and total output of wheat in my country accounted for 20.4% and 20.1% of the total sown area and total output of food crops, respectively. Therefore, it is of great significance to prevent and control wheat diseases and insect pests and increase wheat production and income to ensure the development of national economy.

Contents of the invention

The object of the present invention is to provide an application of Trichoderma aculeatus and a plant growth-promoting bacterial agent.

In order to achieve the above object, the technical solution adopted by the present invention is:

An application of Trichoderma aculeatus, the application of Trichoderma aculeatus in promoting plant growth; wherein, Trichoderma asperelloides has a preservation number of CGMCC No.19914, and a preservation date of June 9, 2020. The preservation unit is the General Microbiology Center of China Microbiological Culture Collection Management Committee, address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and is named as Trichoderma asperelloides (Trichoderma asperelloides).

A plant growth-promoting bacterium agent, which contains the Trichoderma aculeatus.

The number of viable bacteria of Trichoderma aculeatus in the bacterial agent is 10 ⁸ -10 ¹⁰ CFU/g.

The bacterial agent is obtained by adding the activated Trichoderma aculeatus into a solid medium for fermentation and culture.

The fermentation culture condition is 5-40°C, preferably 20-30°C, and cultured for 3-40 days, preferably 5-15 days.

The solid medium is agricultural waste and/or by-products of agricultural product processing; or, a solid medium containing agricultural waste and/or by-products of agricultural product processing.

When the solid medium is agricultural waste and/or agricultural product processing by-products, the agricultural waste and/or agricultural product processing by-products are mixed with water to prepare the medium until the medium is completely wet;

When the solid medium is a solid medium containing agricultural waste and/or by-products of agricultural product processing, the agricultural waste and/or by-products of agricultural product processing are mixed with water to prepare the medium until the medium is completely wet, and nutrients such as glucose, sucrose, peptone, and yeast powder can also be added thereto.

The agricultural waste is one or more of rice husk, crop straw (such as corn, soybean, wheat and other crop straw), peanut shell and corn bran, etc.; the by-products of agricultural product processing are one or more of wheat bran, bean dregs, rice bran, fruit peel, pomace, vegetable dregs and cake.

The solid medium prepared above is prepared by adding one or more of the agricultural waste and/or by-products of agricultural product processing and compounding them with water. The proportion of compounding is adjusted accordingly according to specific needs. Among them, one or more of the agricultural waste and/or by-products of agricultural product processing can be mixed in any proportion, which only provides the energy required for the growth of microorganisms.

The bacterial agent can be powder, suspension or granule.

The invention relates to an application of a bacterial agent, the application of the bacterial agent in promoting plant growth.

The plant is wheat.

The advantages that the present invention has:

The Trichoderma aculeatus in the present invention belongs to Trichoderma and is a kind of beneficial bacteria, which can colonize the root surface of plants and promote the increase of plant roots; the culture medium for making the bacterial agent is agricultural waste or by-products of agricultural product processing, and the material is cheap and easy to obtain. At the same time, it solves the problem of processing these wastes/by-products, is very friendly to the environment, and is conducive to the development of green and environmentally friendly agriculture.

Detailed ways:

Specific embodiments of the present invention will be described in detail below. It should be understood that the specific embodiments described here are only used to illustrate and explain the present invention, and are not intended to limit the present invention.

The bacterial strain adopted in the present invention is Trichoderma, the Trichoderma is isolated from marine pine knotweed, and the fungus is identified as Trichoderma asperelloides. The strain was preserved in the General Microbiology Center of the China Committee for the Preservation of Microbial Strains. The preservation date is June 9, 2020, and the preservation number is CGMCC No.19914.

The microbial inoculum of the present invention is obtained by culturing the Trichoderma in the culture medium, and the amount of thalline contained in the inoculum can be changed in a wide range. For example, in each gram of the microbial inoculum, the number of viable bacteria ^of Trichoderma aculeatus with the preservation number CGMCC No.19914 is ^108-1010 CFU; ^{9-10 10} CFU. During the cultivation process, the concentration of viable bacteria can be obtained by conventional methods, such as blood counting or OD value observation.

The type of the medium can be various mediums that can be used to cultivate Trichoderma aculeatus, for example, the medium is one or more of agricultural waste such as chaff, crop stalks (such as corn, soybean, wheat and other crop stalks), peanut shells and corn bran, etc.; the agricultural product processing by-products are one or more mixed solid medium in wheat bran, bean dregs, rice bran, fruit peels, pomace, vegetable dregs and cakes.

Wherein, the various mediums mentioned above can be sterilized according to conventional sterilization methods and then used for later use, for example, sterilized at 115-125° C. and 1.5-2 standard atmospheric pressure for 10-30 minutes.

Said further that the preparation method of the microbial inoculant can include: inoculating the Trichoderma aculeatus on the potato dextrose agar solid medium (PDA), cultivating in an incubator at 25-28° C. for 5-7 days, then cutting the aforementioned bacterium-containing potato dextrose agar solid medium (PDA) into small pieces and adding it to an Erlenmeyer flask equipped with a solid medium. After large-scale fermentation and culture at 5-40°C for 3-40 days, the culture medium with Trichoderma aculeatus is taken out, dried, and then pulverized with a powder machine to make microbial powder.

The application of the Trichoderma aculeatus or microbial agent in promoting plant growth.

Optionally, the plant is wheat.

Furthermore, Trichoderma aculeatus or the microbial agent containing Trichoderma aculeatus is diluted 200-25000 times with sterile water and used for soaking the seeds for 1-12 hours before sowing.

The present invention is further described in detail below in conjunction with the examples, but the scope of the present invention is not limited to the following examples.

Example 1

Trichoderma aculeatus was inoculated into potato dextrose agar solid medium (PDA), cultured in an incubator at 25°C for 5 days, and then the aforementioned potato dextrose agar solid medium (PDA) containing bacteria was cut into small pieces and added to an Erlenmeyer flask filled with solid medium. After 6 days of large-scale fermentation and culture at 25°C, the bacteria with solid medium were taken out, dried, and ground into powder with a powder machine to make microbial powder. The viable count of Trichoderma aculeatus in the obtained microbial powder was 5×10 ⁹ CFU/g, and this microbial powder was the microbial agent of this embodiment.

The solid medium is a mixture of chaff and wheat bran in a mass ratio of 2:1.

Example 2

Investigation on the growth-promoting effect of Trichoderma aculeatus fungus on wheat.

In this example, the wheat variety Jimai 22 was used as the experimental crop. Select 600 wheat seeds with plump and complete grains, soak them in 75% alcohol for 1 minute to sterilize the seeds, rinse them with sterile water 5-6 times, and then scald them with 50-60°C sterile water for 10 minutes. Take by weighing 0.1g of the Trichoderma aculeatus fungus agent in the above-mentioned embodiment 1, add 100mL sterile water and dilute 1000 times, obtain the agent dilution, soak the wheat seeds with the prepared agent dilution to be the experimental group, soak the wheat seeds with an equal amount of sterile water as the control group, and the soaking time is 1 hour. Place glass petri dishes (1500 mm in diameter) on an ultra-clean workbench, put 2 sheets of filter paper that have been sterilized and fully wet in advance in each petri dish, and place the seeds face up on the filter paper in a single layer with tweezers, and put 100 seeds in each petri dish. There were three repetitions in each of the experimental group and the control group. Cover the petri dish with a lid and place it indoors at a temperature of 26°C. After cultivating to the 7th day, 10 wheat seedlings were randomly selected from each of the experimental group and the control group, and the root number, root length and stem length of the wheat seedlings were recorded, and their average values were recorded (see Table 1).

The number of roots, root length and stem length of seedlings on the seventh day of wheat seed germination when table 1 Class 1 Trichoderma aculeatus agent was diluted 1000 times and soaked for 1 hour

Example 3

In this example, the wheat variety Jimai 22 was used as the experimental crop. Select 600 wheat seeds with plump and complete grains, soak them in 75% alcohol for 1 minute to sterilize the seeds, rinse them with sterile water 5-6 times, and then scald them with 50-60°C sterile water for 10 minutes. Take by weighing 0.02g of the Trichoderma aculeatus fungus agent in the above-mentioned embodiment 1, add 100mL sterile water and dilute 5000 times, obtain the agent dilution, soak the wheat seeds with the prepared agent dilution to be the experimental group, soak the wheat seeds with an equal amount of sterile water as the control group, and the soaking time is 3 hours. Place glass petri dishes (1500 mm in diameter) on an ultra-clean workbench, put 2 sheets of filter paper that have been sterilized and fully wet in advance in each petri dish, and place the seeds face up on the filter paper in a single layer with tweezers, and put 100 seeds in each petri dish. There were three repetitions in each of the experimental group and the control group. Cover the petri dish with a lid and place it indoors at a temperature of 26°C. After cultivating to the 7th day, 10 wheat seedlings were randomly selected from each of the experimental group and the control group, and the root number, root length and stem length of the wheat seedlings were recorded, and their average values were recorded (see Table 2).

Root number, root length and stem length of seedlings on the seventh day of wheat seed germination when table 2 class Trichoderma aculeatus fungal agent was diluted 5000 times and soaked seeds for 3h

Example 4

In this example, the wheat variety Jimai 22 was used as the experimental crop. Select 600 wheat seeds with plump and complete grains, soak them in 75% alcohol for 1 minute to sterilize the seeds, rinse them with sterile water 5-6 times, and then scald them with 50-60°C sterile water for 10 minutes. Take by weighing 0.1g of the Trichoderma aculeatus fungus agent in the above-mentioned embodiment 1, add 100mL concentration and be 3‰ sterile salt (sodium chloride) water and dilute 1000 times, obtain the bacterial agent dilution, soak the wheat seeds with the prepared bacterial agent dilution to be the experimental group, soak the wheat seeds with an equivalent amount of 3‰ sterile saline to be the control group, and the soaking time is 1 hour. Place glass petri dishes (1500 mm in diameter) on an ultra-clean workbench, put 2 sheets of filter paper that have been sterilized and fully wet in advance in each petri dish, and place the seeds face up on the filter paper in a single layer with tweezers, and put 100 seeds in each petri dish. There were three repetitions in each of the experimental group and the control group. Cover the petri dish with a lid and place it indoors at a temperature of 26°C. After cultivating to the 7th day, 10 wheat seedlings were randomly selected from each of the experimental group and the control group, and the root number, root length and stem length of the wheat seedlings were recorded, and their average values were recorded (see Table 3).

Table 3 class Trichoderma aculeatus fungal agent diluted 1000 times 3 ‰ salt water soaking seeds 1h root number, root length and stem length of wheat seed germination on the seventh day

Example 5

In this example, the wheat variety Jimai 22 was used as the experimental crop. Select 600 wheat seeds with plump and complete grains, soak them in 75% alcohol for 1 minute to sterilize the seeds, rinse them with sterile water 5-6 times, and then scald them with 50-60°C sterile water for 10 minutes. Weigh 0.02g of the Trichoderma aculeatus fungus agent in the above-mentioned embodiment 1, add 100mL concentration and be 10‰ sterile saline (sodium chloride) and dilute 5000 times, obtain the inoculant dilution, soak the wheat seeds with the inoculant dilution of configuration as the experimental group, soak the wheat seeds with an equal amount of 10‰ sterile saline as the control group, and the soaking time is 3 hours. Place glass petri dishes (1500 mm in diameter) on an ultra-clean workbench, put 2 sheets of filter paper that have been sterilized and fully wet in advance in each petri dish, and place the seeds face up on the filter paper in a single layer with tweezers, and put 100 seeds in each petri dish. There were three repetitions in each of the experimental group and the control group. Cover the petri dish with a lid and place it indoors at a temperature of 26°C. After cultivating to the 7th day, 10 wheat seedlings were randomly selected from each of the experimental group and the control group, and the root number, root length and stem length of the wheat seedlings were recorded, and their average values were recorded (see Table 4).

Table 4 Trichoderma aculeatus fungal agent diluted 5000 times 10‰ salt water soaking seeds 3h root number, root length and stem length of wheat seed germination on the seventh day

As can be seen from the data in Table 1-4, no matter in fresh water or salt water, compared with the control group in the experimental group, the use of the microbial inoculum containing the Trichoderma aculeatus with the preservation number CGMCC No.19914 in Example 1 has the effect of promoting the growth of wheat, and the use of the microbial inoculant with the Trichoderma aculeatus with the preservation number CGMCC No.19914 in Example 1 can promote rooting of wheat and increase the root number of wheat.

The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited to the specific details in the above-mentioned embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

In addition, it should be noted that the various specific technical features described in the above specific implementation manners may be combined in any suitable manner if there is no contradiction. In order to avoid unnecessary repetition, various possible combinations are not further described in the present invention.

In addition, various combinations of different embodiments of the present invention can also be combined arbitrarily, as long as they do not violate the idea of the present invention, they should also be regarded as the content of the present invention.

Claims (9)

1. An application of trichoderma asperellum, which is characterized in that: application of trichoderma asperellum in promoting rooting of wheat; wherein, the trichoderma asperellum is with a preservation number of CGMCC No.19914, a preservation date of 2020, 6 and 9 days, a preservation unit of China general microbiological culture Collection center, and an address: the classification name of the trichoderma asperellum is Trichoderma asperellum (Trichoderma asperellum) of North Chen Xili No.1, 3 in the Chaoyang area of Beijing cityTrichoderma asperelloides)。

2. The application of a plant growth promoting microbial agent in promoting wheat rooting is characterized in that: the microbial inoculum comprises trichoderma asperellum-like according to claim 1.

3. Use according to claim 2, characterized in that: the number of viable bacteria of the trichoderma asperellum in the microbial inoculum is 10 ⁸ -10 ¹⁰ CFU/g。

4. Use according to claim 2 or 3, characterized in that: the microbial inoculum is obtained by adding activated trichoderma asperellum into a solid culture medium for fermentation culture.

5. The use according to claim 4, wherein: the fermentation culture condition is 5-40 ℃, and the culture is carried out for 3-40 days.

6. The use according to claim 4, wherein: the solid culture medium is agricultural waste and/or agricultural product processing byproducts.

7. The use according to claim 4, wherein: the solid medium contains agricultural waste and/or agricultural product processing byproducts.

8. Use according to claim 6 or 7, characterized in that: the agricultural waste is one or more of crop straw, peanut shell or corn husk; the agricultural product processing by-product is one or more of wheat bran, bean dreg, rice bran, pericarp, fruit dreg, vegetable dreg or cake dreg.

9. Use according to claim 2, characterized in that: the microbial inoculum is powder, suspension or granule.