Hot pepper seedling culture substrate with functions of strengthening seedlings and increasing yield by taking trichoderma asperellum agent and charcoal as materials

Abstract

The invention belongs to the technical field of seedling raising substrates, and particularly relates to a pepper seedling raising substrate which takes a trichoderma asperellum agent and biochar as materials and has the effects of strengthening seedlings and increasing yield. The trichoderma asperellum agent is prepared by mixing a strain LZ35 fermentation broth and a strain X17 fermentation broth in equal mass, wherein the strain LZ35 has a taxonomic name of Trichoderma asperellum, and the preservation number of the strain LZ35 is GDMCC No: 65053; the strain X17 has the taxonomic name of Bacillus subtilis, and the preservation number of the strain X17 is GDMCC No: 64986. According to the invention, a strain LZ35 and a strain X17 which have no antagonistic action are screened from rhizosphere soil of healthy pepper plants. Fermentation liquor of strains LZ35 and X17, wheat straw biochar and a matrix body are mixed to prepare a seedling culture matrix, and when the seedling culture matrix is applied to hot pepper plug seedling culture, it is found that by adding the trichoderma asperellum agent, the germination potential and germination rate of hot pepper seeds and the survival rate of hot pepper seedling plants can be improved, growth of hot pepper seedlings can be promoted, and support can be provided for hot pepper seedling culture and planting.

Description

Technical Field

[0001] The invention belongs to the technical field of seedling cultivation substrates, and particularly relates to a pepper seedling cultivation substrate which takes a Trichoderma acanthosporium agent and biochar as materials and has the functions of strengthening seedlings and increasing yield. Background Art

[0002] With the continuous improvement of people's consumption level, my country's facility vegetable industry has developed rapidly and has become the main direction of vegetable industry development. At present, the annual output of facility vegetables exceeds 265 million, with an output value of nearly 700 billion yuan. The facility vegetable industry has become an important way for Chinese farmers to increase their income and create wealth. Pepper is one of the main vegetables in my country, with a planting area of ​​827,000 hm2 in 2021. 2 , accounting for about 46% of the world's total output. With the increasing planting area of ​​peppers, it has become an important part of the vegetable industry.

[0003] Seedling cultivation is the primary and key technical link in the vegetable industry. However, with the development of large-scale and modern agricultural production, traditional agricultural breeding methods can no longer meet the needs of current agricultural planting because they cannot guarantee the quality of seedlings. Intensive seedling cultivation, represented by plug tray seedling cultivation, has been widely used. Intensive seedling cultivation is easy to operate, saves time and effort, saves seeds and pesticides, can effectively reduce the harm of pests and diseases, overcome the obstacles of continuous cropping, is easy to manage, and the resulting seedlings are strong. The seedling medium is the basis for the growth of the seedling root system and is the key to intensive seedling cultivation technology. Its quality has a huge impact on the quality of the seedlings. The quality of pepper seedlings has a key impact on their yield and fruit quality. Cultivating strong pepper seedlings is the basic prerequisite for increasing pepper yield and revenue.

[0004] The existing commonly used vegetable seedling substrate is a mixture of peat, perlite and vermiculite, or a mixture of peat and vermiculite. In many cases, it only provides a growth environment for vegetable seedlings and has a single function.

[0005] There are a large number of beneficial microorganisms in the soil, among which the rhizosphere growth-promoting bacteria in the rhizosphere micro-domain soil can promote crop growth and increase yield through direct (activating soil nutrients, producing plant hormones and, inducing the enrichment of beneficial microorganisms, etc.) and indirect (producing antibacterial substances, inducing plant systemic resistance and stress, nutrient space competition, etc.) growth-promoting mechanisms, and can effectively alleviate the damage to soil health. For this reason, it is of great significance to study the seedling substrate containing microbial agents.

[0006] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention and should not be regarded as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art. Summary of the invention

[0007] The purpose of the present invention is to provide a pepper seedling raising matrix with the functions of strengthening seedlings and increasing yield, which uses Trichoderma aculeatus and biochar as materials, can improve the germination potential and germination rate of pepper seeds and the survival rate of pepper seedlings, and promote the growth of pepper seedlings.

[0008] To achieve the above object, the present invention provides the following technical solutions:

[0009] A Trichoderma acanthoides agent, wherein the Trichoderma acanthoides agent is a mixture of equal mass of fermentation liquid of strain LZ35 and fermentation liquid of strain X17; wherein,

[0010] The strain LZ35 has a taxonomic name of Trichoderma asperellum, and was deposited in Guangdong Microbiological Culture Collection Center on August 23, 2024, with a deposit number of GDMCC No: 65053;

[0011] The strain X17 has the scientific name Bacillus subtilis, and was deposited in Guangdong Provincial Microbiological Culture Collection Center on August 7, 2024, with the deposit number GDMCC No: 64986.

[0012] Preferably, the preparation method of the fermentation broth of the strain LZ35 is as follows: the strain LZ35 is inoculated on a PDA medium, cultured at a constant temperature of 28°C until spores are produced, spores are washed with sterile water, the bacteria are filtered through 4 layers of gauze, and then the spore concentration is diluted to 1.0×10 7 cfu / g.

[0013] Preferably, the preparation method of the fermentation broth of strain X17 is as follows: strain X17 is inoculated into LB liquid culture medium, placed on a shaker at 37°C and 180 rpm / min and cultured until the logarithmic growth phase, and the bacterial concentration is diluted to 1.0×10 8 cfu / g.

[0014] The invention also provides a seedling cultivation matrix, which contains the Trichoderma acanthosporium agent.

[0015] Preferably, the seedling culture matrix is ​​formed by mixing a matrix body, biochar and Trichoderma aspergillus agent in a mass ratio of 1:0.1:0.4-0.5.

[0016] Preferably, the preparation method of the biochar is: take wheat straw, wash it and dry it to constant weight, crush it into small pieces of 3-5 cm, place it in a carbonization furnace, anaerobic pyrolysis at 450-600°C for 2-3 hours, naturally cool it to room temperature, rinse it with clean water and dry it to constant weight, crush it through a 150-200 mesh sieve to obtain the biochar.

[0017] Preferably, the matrix is ​​formed by mixing peat soil and vermiculite in a mass ratio of 7:3.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] The present invention separates and screens strains from rhizosphere soil of healthy pepper plants, performs a plate confrontation test on strains that grow well, and screens strains LZ35 and strain X17 that have no antagonistic effect between the two strains. Through molecular biological identification, strain LZ35 is Trichoderma aculeatus, and strain X17 is Bacillus subtilis. Fermentation liquid of strains LZ35 and X17, wheat straw biochar and matrix are mixed to prepare a seedling matrix, and when applied to pepper plug seedling cultivation, it is found that adding the Trichoderma aculeatus agent of the present invention can improve the germination potential, germination rate and survival rate of pepper seedlings, promote the growth of pepper seedlings, and provide support for pepper seedling cultivation and planting.

[0020] Collection Information

[0021] The strain LZ35, with the taxonomic name Trichoderma asperellum LZ35, was deposited in Guangdong Microbiological Culture Collection Center on August 23, 2024, with the deposit address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, and the deposit number is GDMCC No: 65053;

[0022] Strain X17, taxonomic name Bacillus subtilis X17, was deposited in Guangdong Provincial Microbiological Culture Collection Center on August 7, 2024. The deposit address is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou City, and the deposit number is GDMCC No: 64986. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 The colony and phylogenetic tree of strain LZ35;

[0024] Figure 2 The colony and phylogenetic tree of strain X17. DETAILED DESCRIPTION

[0025] The following is a clear and complete description of the technical solution of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by technicians in this field without creative work are within the scope of protection of the present invention.

[0026] In the following examples, the PDA medium formula is: 200 g potato, 20 g glucose, 15 g agar, 1000 mL sterile water, pH 7.0.

[0027] The formula of LB liquid culture medium is: 10 g tryptone, 5 g yeast extract, 10 g sodium chloride, 1000 mL sterile water, pH 7.0.

[0028] 1. Isolation and purification of strains

[0029] Rhizosphere soil of healthy pepper plants was collected and prepared into soil suspension. The gradient dilution plate method was used to spread 0.1 mL 10 -3 , 10 -4 , 10 -5 The dilutions were used to screen fungi on PDA medium, with 3 replicates for each gradient, and cultured in a 28°C constant temperature incubator for 3-10 days. -5 , 10 -6 , 10 -7 The dilutions were plated on LB medium to screen bacteria, with 3 replicates per gradient, and placed in a 37°C constant temperature incubator for 1-2 days. The fungi and bacteria that grew well were then isolated, purified, cultured, and numbered.

[0030] 2. Screening and identification of fungi and bacteria

[0031] The isolated and purified strains with good growth were subjected to plate confrontation test to screen out the strains with no antagonistic effect between each other. The DNA was extracted using PowerSoil DNA kit (Mobio Laboratories, Carlsbad, CA, USA), and the taxonomic status of the strains was clarified by 16S / ITS rDNA amplicon sequencing.

[0032] After screening, one fungus strain LZ35 and one bacterial strain X17 were obtained. Molecular biological identification showed that the two strains were: strain LZ35 was Trichoderma asperellum, and strain X17 was Bacillus subtilis. The morphological characteristics and phylogenetic tree of the two strains are shown in Figure 1 and 2 The base sequences are shown in the sequence listings SEQ ID No.1 and SEQ ID No.2.

[0033] 3. Preparation of LZ35 and X17 fermentation broth

[0034] The strain LZ35 was inoculated on PDA medium and cultured at 28°C until spores were produced. The spores were washed with sterile water, filtered through 4 layers of gauze, and then diluted with sterile water to a spore concentration of 1.0×107 cfu / g.

[0035] The strain X17 was inoculated into LB liquid medium and cultured on a shaker at 37°C and 180 rpm / min until the logarithmic growth phase. The bacterial concentration was diluted to 1.0×10 8 cfu / g.

[0036] 4. Effects of different seedling medium on pepper plug seedling cultivation

[0037] Prepare the seedling medium in Table 1, mix the seedling medium evenly and put it into a 72-hole tray. In Table 1, the matrix is ​​made of peat soil and vermiculite mixed in a mass ratio of 7:3; the preparation method of biochar is as follows: take wheat straw, wash it, dry it to constant weight, crush it into small pieces of 5 cm, place it in a carbonization furnace, pyrolyze it at 00℃ for 2.5h, cool it naturally to room temperature, rinse it with clean water, dry it to constant weight, crush it through a 150-mesh sieve, and obtain the biochar;

[0038] Table 1 Different seedling medium

[0039] Element Seedling medium 1 Seedling medium 2 Seedling medium 3 Seedling medium 4 Matrix body 2000g 2000g 2000g 2000g Biochar 200g 200g 200g 200g Strain LZ35 fermentation broth -- 800g -- 400g Strain X17 fermentation broth -- -- 800g 400g Sterile water 800g -- -- --

[0040] Take the uniform and full "Tian Deng Zhi Tian Jiao" pepper seeds, disinfect them with 75% ethanol for 2 minutes, rinse them with sterile water 3 times, take them out and soak them in sterile water for 24 hours. Sow the treated pepper seeds in 72-hole trays, cover them with 0.5 cm of seedling medium after sowing, sow 1 seed per hole, repeat 4 times for each seedling medium, and repeat 25 pepper seeds per time. Record the number of germinations on the 7th day and the 14th day, and measure the number of plants, plant height, stem diameter, fresh weight and dry weight on the 50th day, and calculate the germination potential, seed germination rate, survival rate, leaf area and seedling index. The results are shown in Table 2.

[0041] in,

[0042]

[0043]

[0044]

[0045]

[0046] Table 2 Effects of different seedling medium on pepper seedling tray cultivation

[0047] Measurement indicators Seedling medium 1 Seedling medium 2 Seedling medium 3 Seedling medium 4 Number of seeds germinated and broke through the soil on the 7th day 6 14 11 27 Number of germinated seeds on the 14th day 79 91 84 97 Number of surviving seedlings on the 50th day (plants) 68 84 79 93 Plant height (cm) 9.34 12.08 10.13 18.99 Stem diameter (mm) 1.93 2.49 2.65 4.07 Fresh weight (g) 2.09 9.84 7.50 21.11 Dry weight (g) 0.14 0.23 0.21 0.46 Germination potential (%) 6 14 11 27 Seed germination rate (%) 79 91 84 97 Survival rate (%) 68 84 79 93 Seedling index 0.0289 0.0474 0.0549 0.0986

[0048] As shown in Table 1, compared with the seedling matrix 1, the plant height, fresh weight, number of leaves and germination potential, seed germination rate, survival rate and seedling index of the seedling matrix 2-4 are significantly improved, and the improvement of the seedling matrix 4 in all aspects is significantly better than that of the seedling matrix 2 and the seedling matrix 3, and the difference is significant. On the whole, adding the Trichoderma acanthosporum agent of the present invention to the matrix body can significantly promote the germination of pepper seeds and the growth of seedlings.

[0049] In summary, adding the Trichoderma spinulosum agent of the present invention into the matrix can improve the germination potential and germination rate of pepper seeds and the survival rate of pepper seedlings, and promote the growth of pepper seedlings.

[0050] The foregoing description of specific exemplary embodiments of the present invention is for the purpose of illustration and demonstration. These descriptions are not intended to limit the present invention to the precise form disclosed, and it is clear that many changes and variations can be made based on the above teachings. The purpose of selecting and describing the exemplary embodiments is to explain the specific principles of the present invention and its practical application, so that those skilled in the art can realize and utilize various different exemplary embodiments of the present invention and various different selections and changes. The scope of the present invention is intended to be limited by the claims and their equivalents.

Claims

1. A Trichoderma acanthosporium agent, characterized in that The Trichoderma spinulosa agent is a mixture of equal amounts of fermentation broth of strain LZ35 and fermentation broth of strain X17; wherein, The strain LZ35 has a taxonomic name of Trichoderma asperellum, and was deposited in Guangdong Microbiological Culture Collection Center on August 23, 2024, with a deposit number of GDMCC No: 65053; The strain X17 has the scientific name Bacillus subtilis, and was deposited in Guangdong Provincial Microbiological Culture Collection Center on August 7, 2024, with the deposit number GDMCC No: 64986.

2. The Trichoderma spinulosa agent according to claim 1, characterized in that The preparation method of the fermentation liquid of the strain LZ35 is as follows: the strain LZ35 is inoculated on a PDA medium, cultured at a constant temperature of 28°C until spores are produced, spores are washed off with sterile water, the bacteria are filtered through 4 layers of gauze, and then the spore concentration is diluted to 1.0×10 7 cfu / g.

3. The Trichoderma aspergillus agent according to claim 1, characterized in that The preparation method of the fermentation broth of strain X17 is as follows: strain X17 is inoculated into LB liquid culture medium, placed on a shaker at 37°C and 180 rpm / min and cultured until the logarithmic growth phase, and the bacterial concentration is diluted to 1.0×10 8 cfu / g.

4. A seedling culture medium, characterized in that: The seedling culture medium comprises the Trichoderma aspergillus agent according to claim 1.

5. The seedling raising matrix according to claim 4, characterized in that The seedling culture matrix is ​​formed by mixing a matrix body, biochar and a Trichoderma spinulosa agent in a mass ratio of 1:0.1:0.4-0.

5.

6. The seedling culture medium according to claim 5, characterized in that: The preparation method of the biochar is as follows: wheat straw is taken, washed and dried to constant weight, crushed into small pieces of 3-5 cm, placed in a carbonization furnace, anaerobic cracked at 450-600° C. for 2-3 hours, naturally cooled to room temperature, rinsed with clean water and dried to constant weight, crushed to pass through a 150-200 mesh sieve, and the biochar is obtained.

7. The seedling culture medium according to claim 5, characterized in that: The matrix is ​​formed by mixing peat soil and vermiculite in a mass ratio of 7:3.

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