CN116724855A - Strawberry seedling and cultivation matrix formula containing trichoderma and bacillus and application thereof - Google Patents

Abstract

The application discloses a formula and application of a strawberry seedling culture and cultivation substrate containing trichoderma and bacillus, wherein the strawberry seedling culture and cultivation substrate is obtained by inoculating trichoderma Guizhou NJAU 4742 solid strain or/and SQR9 solid strain to a basic substrate, is an optimal product in the strawberry seedling culture substrate, and is a novel substrate integrating organic fertilizer and functional microorganisms, not only can be used in a pot plant, but also can be used in a strawberry overhead plant, and different substrates are adopted in different periods of strawberries, so that the growth, development and yield of strawberries are effectively promoted.

Description

A strawberry seedling and cultivation substrate formula containing Trichoderma and Bacillus and its application

Technical field

The invention belongs to the field of biotechnology, specifically relates to the technical field of intensive agricultural production, and more specifically relates to a strawberry seedling and cultivation matrix formula containing Trichoderma and Bacillus and its application.

Background technique

Strawberries are bright in color, soft and juicy, rich in fragrance, sweet and sour. In addition to being rich in vitamins and minerals, they also contain glucose, citric acid, carotene, riboflavin, etc. They are a popular seasonal fruit. Strawberry fruit has delicate flesh and high nutritional value. It is a fruit with extremely high economic benefits. In order to solve a series of problems such as the large area and high labor intensity of current strawberry seedling production, the elevated layered seedling device developed by the Facility Agriculture Research Institute of the Planning and Design Institute of the Ministry of Agriculture is used in planting. This device has the characteristics of high land utilization rate and labor intensity. It has the characteristics of low intensity, simple seedling raising technology, high breeding coefficient and high seedling quality. A large number of strawberry seedlings can be put on the market in a short period of time, but elevated strawberry seedlings are currently mostly grown on substrates.

At present, some scholars have done some research on strawberry-specific elevated substrates. The CN106045791A invention discloses an elevated strawberry-specific substrate that is conducive to a balanced and continuous supply of water and fertilizer for strawberries. The CN111602581A invention discloses an elevated strawberry substrate made of peat, vermiculite, perlite, and biological A special strawberry substrate that uses charcoal as the main raw material to increase strawberry yield and quality. The CN108849390A invention discloses a special matrix for strawberry facilities using Agaricus bisporus residue as raw material, but these inventions do not take into account the growth and reproduction of strawberry sub-strains and the application of beneficial microorganisms in the matrix.

Contents of the invention

The object of the present invention is to provide a strawberry seedling and cultivation substrate formula containing Trichoderma and Bacillus and its application. The strawberry seedlings and cultivation substrate formula containing Trichoderma and Bacillus in the present invention have significant effects in promoting strawberry growth, increasing strawberry yield, and improving the physical and chemical properties of the substrate.

The purpose of the present invention is mainly implemented through the following technical solutions: a strawberry seedling and cultivation substrate containing Trichoderma and Bacillus is obtained through the following steps: inoculating Guizhou Trichoderma NJAU 4742 solid strain or/and SQR9 solid strain into the base Obtained after matrix.

The Guizhou Trichoderma strain NJAU 4742 of the present invention, whose deposit number is CGMCC NO. 12166, is an existing published strain (as disclosed in patent 201810768488.0).

Bacillus amyloliquefaciens SQR9 of the present invention, whose deposit number is CGMCC NO. 5808, is a currently disclosed strain.

The Guizhou Trichoderma NJAU 4742 solid strain of the present invention can be obtained by solid fermentation according to conventional methods in this field. The present invention provides a specific method:

① Inoculate Guizhou Trichoderma NJAU 4742 into PDA liquid culture medium for liquid fermentation to prepare Trichoderma liquid strain;

② The solid fermentation material is inoculated with Trichoderma liquid strain according to 0.05-0.15ml/g. The thickness of the plate fermentation material is 2-4cm, the temperature is 26-28°C, and the humidity is 70-90%. Cultivate for 3-5 days to obtain the solid strain of Trichoderma. .

The PDA liquid culture medium of the present invention can be a conventional PDA liquid culture medium purchased in the market. The inoculation amount can be according to the conventional inoculation amount in this field, for example, 200 ml liquid culture medium can be inoculated with 3-5 Trichoderma blocks with a diameter of 0.3-0.7 cm.

In some embodiments, the conditions for liquid fermentation in step ① are pH 6.8-7.2, temperature 25-30°C, 150-200 rpm, and time 4-5 days.

In some embodiments, the solid fermentation material raw materials in step ② are corn straw and rice bran with a mass ratio of 0.8 to 1.2:1, a moisture content of 50% to 70%, and a pH of 2.5-3.0. The solid fermentation material raw materials can be sterilized by conventional methods, such as sterilization at 115°C for 40 minutes. The pH can be adjusted by conventional methods. In a preferred embodiment, acidolyzed amino acids are used for adjustment.

The SQR9 solid strain of the present invention can be obtained by solid inoculation according to conventional methods in this field. The present invention provides a specific method:

① Inoculate SQR9 strain into LB liquid culture medium for liquid fermentation to prepare SQR9 liquid strain;

② Inoculate the SQR9 liquid strain prepared in step ① into the solid material at a rate of 0.05 to 0.15ml/g to obtain the SQR9 solid strain.

The LB liquid culture medium formula of the present invention can be a conventional formula in this field, for example, peptone 10g/L (imported), yeast powder 5g/L (imported), sodium chloride 5g/L (analytically pure), pH 7.0 . The inoculation amount can be according to the conventional inoculation amount in this field, for example, 200 ml of liquid culture medium is infused with 3-5 SQR9 bacterial blocks with a diameter of 0.3-0.7 cm.

In some embodiments, the conditions for step ① SQR9 liquid fermentation of the present invention are pH 6.8-7.2, temperature 25-30°C, 150-200 rpm, and time 2-3 days.

In some embodiments, the solid material raw materials in step ② are corn straw and rice bran with a mass ratio of 0.8 to 1.2:1.

In a specific embodiment, the Guizhou Trichoderma NJAU 4742 solid strain of the present invention is inoculated into the basic matrix at a ratio of 25 to 35 kg/ton.

In a specific embodiment, the SQR9 solid bacterial strain of the present invention is inoculated into the basic matrix at a ratio of 25 to 35 kg/ton.

The invention also provides the application of the strawberry seedlings and cultivation substrate containing Trichoderma and Bacillus in strawberry seedlings and cultivation.

The present invention also provides a method for raising and cultivating strawberry seedlings. The method specifically uses substrate one for cultivation during the elevated seedling cultivation period of strawberry. After obtaining the sub-strains, the sub-strains are cultivated using substrate two, and after two months, substrate three is used for cultivation. The present invention The first substrate is the strawberry seedling and cultivation substrate containing Trichoderma and Bacillus of the present invention plus 10% organic fertilizer by weight; the second substrate is a basic substrate plus 10% organic fertilizer by weight. Or the second substrate is the strawberry seedling and cultivation substrate containing Trichoderma and Bacillus of the present invention plus 10% organic fertilizer by weight; the third substrate is the basic substrate plus 10% organic fertilizer by weight, or the substrate The third step is the strawberry seedling cultivation and cultivation substrate containing Trichoderma and Bacillus according to the present invention plus 10% by weight of organic fertilizer and 10% by weight of soil.

In some embodiments, the basic matrix of the present invention is vermiculite, perlite, and woody peat mixed in a ratio of 3:3:1 by volume.

The soil described in the present invention is any soil suitable for strawberry growth.

The invention also provides the application of the strawberry seedlings and cultivation substrates containing Bacillus and Trichoderma in promoting strawberry growth, increasing strawberry yield and improving the physical and chemical properties of the substrate.

Compared with strawberry seedling raising and cultivation substrate products currently on the market, the present invention has the following advantages:

1) It is the best product among strawberry seedling cultivation substrates, a new substrate integrating organic fertilizer and functional microorganisms.

2) Compared with ordinary substrates, strawberry-specific seedling substrates can provide organic nutrients for strawberries, permanently improve substrate fertility, and are of great significance to the sustainable utilization of substrates and ecological environment protection.

3) The special strawberry seedling cultivation substrate of the present invention has the advantage of lower cost.

4) This product can significantly increase the dry weight of the above-ground and underground parts of strawberry; in addition, Guizhou Trichoderma strain NJAU4742 and Bacillus amyloliquefaciens SQR9 can also regulate Jiankang soil microflora. The product of the present invention can promote strawberry growth as a whole.

5) The substrate prepared by the method of the present invention can be used not only in potted plants, but also in strawberry elevated stands, both of which can achieve the effect of promoting strawberry growth.

6) By using different substrates at different stages of strawberry, the growth, development and yield of strawberry can be effectively promoted.

Description of drawings

Figure 1 shows the above-ground dry weight of strawberry plants in Example 3.

Figure 2 is the dry weight of the underground part of the strawberry plant in Example 3.

Figure 3 shows the nitrate nitrogen content of the matrix in Example 3.

Figure 4 shows the ammonium nitrogen content of the matrix in Example 3.

Figure 5 shows the total phosphorus content of the matrix in Example 3.

Figure 6 shows the total potassium content of the matrix in Example 3.

Detailed ways

The following examples facilitate a better understanding of the present invention, but do not limit the present invention. The experimental methods in the following examples are all conventional methods unless otherwise specified. The test materials used in the following examples were all purchased from conventional biochemical reagent stores unless otherwise specified.

Unless otherwise specified, the basic matrix used in the following examples is vermiculite, perlite, and woody peat mixed evenly in a volume ratio of 3:3:1.

Unless otherwise specified, the organic fertilizer described in the present invention is made from aerobic composting of agricultural organic waste according to conventional methods in this field. The waste is mainly livestock and poultry manure, straw, tail vegetables, etc.

Unless otherwise specified, “Trichoderma” in the following examples refers to Trichoderma guizhouense NJAU 4742.

Example 1 Preparation of Guizhou Trichoderma NJAU 4742 solid strain

(1) Trichoderma strain liquid fermentation: Inoculate Trichoderma into PDA liquid culture medium for liquid fermentation to prepare Trichoderma liquid strain. The fermentation conditions are pH 7.0, temperature 28°C, 180 rpm, time 5 days, that is, Trichoderma liquid strain Species (including mycelium and sporophyte);

(2) Trichoderma species solid fermentation: The solid fermentation raw material is 50% corn straw + 50% rice bran. The pH of the material is adjusted to 2.5-3.0 through acidolysis of amino acids. The water content is adjusted to 60% with tap water. The solid sterilization tank is sterilized at 115°C. After 40 minutes, the material is cooled and inoculated with 10% Trichoderma liquid strain. The thickness of the plate fermentation material is 3cm, the temperature is 28-30°C, and the humidity is 70-90%. It is cultured in a class 10,000 clean room for 4 days. The number of Trichoderma spores is higher than 5×10 ^9cfu /g.

When used, it is inoculated into the basic matrix at a weight percentage of 3%.

Example 2 Preparation of SQR9 solid bacterial strain

(1) Trichoderma strain liquid fermentation: Inoculate SQR9 bacteria into LB liquid culture medium for liquid fermentation to prepare SQR9 liquid strain. The fermentation conditions are pH 7.0, temperature 28°C, 180 rpm, and time of 3 days, that is, SQR9 liquid strain;

(2) SQR9 strain solid fermentation: The solid fermentation raw material is 50% corn straw + 50% rice bran, and the SQR9 liquid strain is adsorbed to the material according to 10% inoculation amount, and the pH is 7.0.

When used, it is inoculated into the basic matrix at a weight percentage of 3%.

Example 3 Elevated seedling cultivation test

The experiment was conducted at the Golden Manor strawberry base. The experiment was designed with 5 treatments. On the basis of a 3:3:1 mixture of perlite: vermiculite: wood peat, each matrix was added with 10% amino acid organic fertilizer. 3 different addition examples were designed. 1. Treatment of Guizhou Trichoderma NJAU 4742 solid strain or Example 2 SQR9 solid strain combination, and at the same time, the matrix without adding any solid strain was used as a blank control. A total of 4 processes are set. ①CK: No added strains ②NJAU 4742: Added Trichoderma ③SQR9: Added SQR9

④N+S: Add SQR9 and Trichoderma.

Table 1 Effects of different treatments on strawberry crop biomass and soil physical and chemical properties

Note: Different letters represent significant differences between different treatments, P<0.05.

As shown in Figure 1 and Table 1, the substrate adding SQR9 and Trichoderma can significantly promote the growth of the aboveground part of strawberry. Compared with CK, the dry weight of the aboveground part of the substrate adding SQR9 and Trichoderma increased by 77%, as shown in Figure 2 It shows that the substrate adding SQR9 and Trichoderma can also significantly promote the growth of the underground part of strawberry. Compared with CK, the dry weight of the underground part of the substrate adding SQR9 and Trichoderma increased by 79%, as shown in Figure 3 and Figure 4. The ammonium nitrate nitrogen content of the matrix of SQR9 and Trichoderma increased significantly. Compared with CK, the nitrate nitrogen of the matrix of SQR9 and Trichoderma increased by 70%, and the ammonium nitrogen content increased by 107.4%, as shown in the figure. As shown in Figure 5, the total phosphorus content of the matrix added with SQR9 and Trichoderma has been improved to a certain extent, and it has increased by 12% compared with CK. As shown in Figure 6, the total potassium content of the matrix added with SQR9 and Trichoderma has also been improved to a certain extent, compared with CK. compared to an increase of 20.6%.

In summary, the strawberry-specific seedling substrate containing Trichoderma and Bacillus can improve the physical and chemical properties of the substrate during strawberry seedling cultivation and promote strawberry growth.

Example 4 Potted test on seedling root promotion

The test was arranged at the Yixing test base. During the strawberry rooting period, the test set up 64 treatments. Perlite: vermiculite: woody peat was based on 3:3:1. Different proportions of soil and different contents of organic fertilizers were added on the basis. And different combinations of Trichoderma Guizhou NJAU 4742 solid strain in Example 1 or SQR9 solid strain in Example 2, a total of 3 different factors, each factor has a total of 4 different treatments, and each factor is matched with each other, for a total of 64 treatments , different soil proportions are divided into A0 (0% soil), A1 (15% soil), A2 (30% soil), A3 (45% soil); different contents of organic fertilizer are divided into B0 (0% organic fertilizer), B1 ( 2.5% organic fertilizer), B2 (5% organic fertilizer), B3 (10% organic fertilizer); adding different microbial strains is divided into C0 (no added strains), C1 (added SQR9), C2 (added Trichoderma), C3 (SQR9+Trichoderma added). Each treatment was repeated 6 times, and strawberry crop biomass was calculated 2 months after transplanting the seedlings.

Table 2 Effects of different substrate treatments on strawberry crop biomass

It can be seen from Table 2 that during the rooting stage of strawberry seedlings, the substrates A0B3C2, A0B3C1, A0B3C0, and A0B3C3 can best promote the growth of strawberries, that is, the soil proportion content is 0%, the organic fertilizer content is 10%, Trichoderma is added, SQR9 is added, or no microorganisms are added , has a very good effect on the rooting of seedlings.

Example 5 Elevated cultivation test

The test was conducted at the Golden Manor strawberry base. Each treatment used an elevated rack with a length of 17m, and each elevated rack was 20cm wide. The yield described in the test was the yield of the elevated rack with an area of ^3.4m2 . A total of 4 treatments were designed in the experiment, in which the basic matrix was a matrix of perlite: vermiculite: woody peat mixed in a ratio of 3:3:1, and Nannong matrix was the basic matrix with 10% by weight of amino acid organic fertilizer added. The experimental processing is as follows:

①T1: Golden Manor commercial matrix ②T2: Nannong matrix ③T3: Nannong matrix + 10% soil ④ T4: Nannong matrix + 10% soil + Trichoderma + SQR9.

Table 3 Effects of different treatments on strawberry yield

As shown in Table 3, the yield of Nannong substrate increased by 26.2% compared with the commercial substrate used in the Golden Manor base. The Nannong substrate added 10% soil increased the yield by 50.9% compared with the commercial substrate. The Nannong substrate added 10% soil and Trichoderma and SQR9 increased the yield by 61.5% compared to the commercial substrate.

In summary, adding 10% soil to Nannong's matrix formula, and adding Trichoderma and SQR9 to the matrix can increase strawberry yield.

In summary, it can be seen that this application has achieved good growth, development and yield improvement effects by using different substrates for strawberries at different stages.

Claims (10)

1. A strawberry seedling culture and cultivation substrate containing trichoderma and bacillus is characterized in that the substrate is obtained by inoculating trichoderma Guizhou NJAU 4742 solid strain or/and SQR9 solid strain to a basic substrate.

2. The strawberry seedling culture and cultivation medium according to claim 1, wherein the preparation method of the trichoderma guizhou NJAU 4742 solid strain comprises the following steps:

(1) inoculating trichoderma Guizhou NJAU 4742 into a PDA liquid culture medium for liquid fermentation to prepare trichoderma liquid strains;

(2) inoculating the solid fermentation material with trichoderma liquid strain in 0.05-0.15 ml/g, and culturing at 26-28 deg.c and 70-90% humidity for 3-5 days to obtain trichoderma solid strain.

3. The strawberry seedling and cultivation medium according to claim 2, wherein the liquid fermentation condition in the step (1) is pH 6.8-7.2, the temperature is 25-30 ℃, the speed is 150-200 rpm, and the time is 4-5 days; the solid fermentation material in the step (2) comprises the following raw materials in percentage by mass of 0.8-1.2: 1, corn stalk and rice bran with water content of 50-70% and pH of 2.5-3.0.

4. The strawberry seedling and cultivation medium according to claim 1, wherein the preparation method of the SQR9 solid strain is as follows:

(1) inoculating SQR9 strain into LB liquid culture medium for liquid fermentation to prepare SQR9 liquid strain;

(2) inoculating the SQR9 liquid strain prepared in the step (1) to a solid material according to the ratio of 0.05-0.15 ml/g to obtain the SQR9 solid strain.

5. The strawberry seedling and cultivation medium according to claim 4, wherein the SQR9 liquid fermentation condition of the step (1) is pH 6.8-7.2, the temperature is 25-30 ℃, the speed is 150-200 rpm, and the time is 2-3 days;

the solid material raw material in the step (2) is 0.8-1.2 in mass ratio: 1 corn stalks and rice bran.

6. The strawberry seedling and cultivation medium according to claim 1, wherein the trichoderma Guizhou NJAU 4742 solid strain is inoculated into the basic medium according to the proportion of 25-35 kg/ton; the SQR9 solid strain is inoculated into the basic matrix according to the proportion of 25-35 kg/ton.

7. The strawberry seedling and cultivation medium according to claim 1, wherein the basic medium is vermiculite, perlite and wood peat according to volume 3:3: 1.

8. The use of the strawberry seedling and cultivation medium according to any one of claims 1-7 in strawberry seedling and cultivation.

9. Use of the strawberry seedling and cultivation substrate of any one of claims 1-7 for promoting strawberry growth, increasing strawberry yield and improving physical and chemical properties of the substrate.

10. A strawberry seedling and cultivation method specifically comprises the steps of culturing strawberry in an overhead seedling stage by adopting a first substrate, culturing the obtained seed strain by adopting a second substrate, and culturing the seed strain by adopting a third substrate after 2 months; the first substrate is the strawberry seedling raising and cultivating substrate according to any one of claims 1-7, and the weight percentage of the first substrate is 10% of organic fertilizer; adding 10% of organic fertilizer by weight into a base matrix serving as a second matrix, or adding 10% of organic fertilizer by weight into a strawberry seedling raising and cultivating matrix according to any one of claims 1-7; the third substrate is a basic substrate added with 10 percent of organic fertilizer by weight, or the third substrate is a strawberry seedling raising and cultivation substrate added with 10 percent of organic fertilizer by weight according to any one of claims 1 to 7, and then added with 10 percent of soil by weight.