Preparation and application of bacterial strain for accelerating growth speed of agaricus bisporus based on AbChi1 and AbCbm1 gene co-silencing

Abstract

The invention provides preparation and application of a bacterial strain for increasing the growth speed of agaricus bisporus based on AbChi1 and AbCbm1 gene co-silencing, and particularly belongs to the technical field of gene engineering. The invention relates to an application of a co-silent AbChi1 gene and an AbCbm1 gene in improving the growth speed of agaricus bisporus mycelia. The number of the AbChi1 gene is AGABI2DRAFT211936, and the number of the AbCbm1 gene is AGABI2DRAFT193505, and the number of the AbCbm1 gene is AGABI2DRAFT211936. The AbChi1 gene and the AbCbm1 gene are co-silenced in the agaricus bisporus, so that the mycelial growth speed of the agaricus bisporus can be increased, the yield of the agaricus bisporus is further increased, and the economic benefit is improved.

Description

Technical Field

[0001] The invention belongs to the technical field of genetic engineering, and particularly relates to application of co-silencing AbChi1 and AbCbm1 genes in improving the growth rate of Agaricus bisporus mycelium. Background Art

[0002] Agaricus bisporus (Large) Sing. is a fungus of the Agaricus family and the Agaricus genus, also known as white mushroom, mushroom, and foreign mushroom. Agaricus bisporus is edible and delicious. It is an edible fungus with a large cultivation scale and a wide cultivation range. Its protein content is as high as 42% (dry weight), and it is rich in amino acids, nucleotides, and vitamins. Agaricus bisporus can also be used as medicine. Under wild conditions, Agaricus bisporus grows mostly in grasslands, pastures, and composts in spring, summer, and autumn. With the development of artificial cultivation technology, artificial cultivation has spread to most provinces and regions in China. Promoting the growth of Agaricus bisporus hyphae has a good driving effect on increasing the yield of Agaricus bisporus, enhancing disease resistance, and improving economic benefits. However, at present, the measures to promote the growth of Agaricus bisporus hyphae mainly include controlling the appropriate temperature, humidity, ventilation, and preventing diseases and insect pests. At present, there is no effective method based on genetic engineering technology to promote the growth of Agaricus bisporus. Summary of the invention

[0003] The purpose of the present invention is to provide a preparation and application of a strain for accelerating the growth rate of Agaricus bisporus based on co-silencing of AbChi1 and AbCbm1 genes. Co-silencing of AbChi1 gene and AbCbm1 gene in Agaricus bisporus can increase the growth rate of Agaricus bisporus mycelium, thereby increasing the yield of Agaricus bisporus and improving economic benefits.

[0004] The present invention provides the use of co-silencing AbChi1 gene and AbCbm1 gene in improving the growth rate of Agaricus bisporus mycelium; the AbChi1 gene is numbered AGABI2DRAFT_211936, and the AbCbm1 gene is numbered AGABI2DRAFT_193505.

[0005] The present invention also provides the use of co-silencing AbChi1 gene and AbCbm1 gene in improving the yield of Agaricus bisporus; the AbChi1 gene is numbered AGABI2DRAFT_211936, and the AbCbm1 gene is numbered AGABI2DRAFT_193505.

[0006] The present invention also provides the use of co-silencing AbChi1 gene and AbCbm1 gene in preparing Agaricus bisporus strain with fast mycelium growth rate; the AbChi1 gene is numbered AGABI2DRAFT_211936, and the AbCbm1 gene is numbered AGABI2DRAFT_193505.

[0007] The present invention also provides the use of co-silencing AbChi1 gene and AbCbm1 gene in shortening the production cycle of Agaricus bisporus; the AbChi1 gene is numbered AGABI2DRAFT_211936, and the AbCbm1 gene is numbered AGABI2DRAFT_193505.

[0008] Preferably, the Agaricus bisporus comprises Agaricus bisporus A 15 .

[0009] Preferably, the reagent used for co-silencing the AbChi1 gene and AbCbm1 gene includes an AbChi1 gene and AbCbm1 gene co-silencing vector.

[0010] The present invention also provides a vector for improving the growth rate of Agaricus bisporus mycelium, wherein the vector is a co-silencing vector of AbChi1 gene and AbCbm1 gene; the number of the AbChi1 gene is AGABI2DRAFT_211936, and the number of the AbCbm1 gene is AGABI2DRAFT_193505.

[0011] Preferably, the vector is based on pCAMBIA1303 as a basic plasmid, containing a GPD promoter and an AbChi1 RNAi fragment expressed by the GPD promoter, and a TEF promoter and an AbCbM1 RNAi fragment expressed by the TEF promoter.

[0012] The present invention also provides a method for constructing a Agaricus bisporus strain with fast mycelium growth rate, comprising the following steps:

[0013] The vector described in the above technical solution is transformed into Agrobacterium to obtain Agrobacterium carrying the vector, and the Agrobacterium carrying the vector is used to infect Agaricus bisporus to obtain an Agaricus bisporus strain with fast mycelium growth rate.

[0014] The present invention also provides an Agaricus bisporus strain with fast mycelium growth rate obtained by the construction method described in the above technical scheme.

[0015] The present invention provides an application of co-silencing AbChi1 and AbCbm1 genes in increasing the growth rate of Agaricus bisporus mycelium. The present invention utilizes gene co-silencing technology to co-silence AbChi1 and AbCbm1 genes in Agaricus bisporus, and has the following beneficial effects:

[0016] 1. Increase the growth rate: Targeted regulation of the AbChi1 gene and the AbCbm1 gene can effectively accelerate the growth rate of mushrooms.

[0017] 2. Increase yield: Regulate the growth rate of mushroom mycelium through co-silencing technology to increase the overall yield of mushrooms.

[0018] 3. Improve the commercial value and market competitiveness of Agaricus bisporus.

[0019] 4. Optimize mycelium growth: By silencing inhibitory genes, the expansion rate of mycelium on the culture medium can be accelerated, the culture and spawning time of the strain can be shortened, thereby speeding up the entire production cycle.

[0020] 5. Support precision breeding: It can achieve precise regulation at the genetic level. Compared with traditional hybrid breeding, it is more efficient and has a shorter cycle, and can quickly obtain new varieties of Agaricus bisporus with excellent traits.

[0021] 6. Reduce production risks: By stabilizing and regulating the AbChi1 gene and the AbCbm1 gene, gene co-silencing technology can improve the stability of mushroom growth and reduce production losses caused by gene mutations or environmental interference.

[0022] The present invention can not only accelerate the production efficiency of mushrooms, but also provide important support for industrial upgrading and personalized product development, and is an important tool for modern agricultural biotechnology. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0024] Figure 1 This is a diagram showing the amplification results of the AbChi1 RNAi fragment provided by the present invention;

[0025] Figure 2 The pcambia1303-FvGPDA-AbCbM1-AbTEF vector map provided by the present invention;

[0026] Figure 3 This is a diagram showing the electrophoresis results of the AbCbm1-AbChi1 RNAi fragment provided by the present invention;

[0027] Figure 4 The pcambia1303-FvGPDA-AbCbM1-AbChi1-AbTEF plasmid image provided by the present invention;

[0028] Figure 5 The electrophoresis diagram of the amplified product of the pcambia1303-FvGPDA-AbCbM1-AbChi1-AbTEF transformant provided by the present invention;

[0029] Figure 6 This is a comparison chart of the growth rates of different types of strains provided by the present invention. DETAILED DESCRIPTION

[0030] The present invention provides an application of co-silencing AbChi1 gene and AbCbm1 gene in improving the growth rate of Agaricus bisporus mycelium; the AbChi1 gene is numbered AGABI2DRAFT_211936, and the AbCbm1 gene is numbered AGABI2DRAFT_193505. In a specific embodiment, the Agaricus bisporus includes Agaricus bisporus A 15 . In a specific embodiment, the reagent used for co-silencing the AbChi1 gene and the AbCbm1 gene includes a co-silencing vector for the AbChi1 gene and the AbCbm1 gene. In the early transcriptome of the present invention, the AGABI2DRAFT_211936 gene in the Agaricus bisporus H97 variety was screened out, and it was cloned in the Agaricus bisporus A15 variety and named AbChi1. gene. AbChi1 is similar to the chitinase gene. The yeast two-hybrid technology was used to screen out the gene AGABI2DRAFT_193505 that interacts with the AGABI2DRAFT_211936 gene, and it was cloned in the Agaricus bisporus A15 variety and named AbCbm1. The gene AbCbm1 is a gene encoding a carbohydrate binding module, which plays a similar role to the AbChi1 gene in some aspects and can enhance the function of AbChi1 to a certain extent. The present invention found that the co-silencing technology is implemented on the chitinase gene AbChi1 and the carbohydrate binding module gene AbCbm1 to regulate the gene expression amount, which can control the growth of Agaricus bisporus, specifically to increase the mycelial growth rate.

[0031] The present invention also provides the use of co-silencing AbChi1 gene and AbCbm1 gene in increasing the yield of Agaricus bisporus; the AbChi1 gene is numbered AGABI2DRAFT_211936, and the AbCbm1 gene is numbered AGABI2DRAFT_193505. In a specific embodiment, the Agaricus bisporus includes Agaricus bisporus A 15 In a specific embodiment, the reagent used for co-silencing the AbChi1 gene and the AbCbm1 gene includes an AbChi1 gene and AbCbm1 gene co-silencing vector. Co-silencing the AbChi1 gene and the AbCbm1 gene can promote the growth of Agaricus bisporus hyphae, thereby increasing the yield of Agaricus bisporus.

[0032] The present invention also provides the use of co-silencing AbChi1 gene and AbCbm1 gene in preparing Agaricus bisporus strain with fast mycelium growth rate; the AbChi1 gene is numbered AGABI2DRAFT_211936, and the AbCbm1 gene is numbered AGABI2DRAFT_193505. In a specific embodiment, the Agaricus bisporus includes Agaricus bisporus A 15 In a specific embodiment, the reagent used for co-silencing the AbChi1 gene and the AbCbm1 gene includes an AbChi1 gene and AbCbm1 gene co-silencing vector.

[0033] The present invention also provides an application of co-silencing AbChi1 gene and AbCbm1 gene in shortening the production cycle of Agaricus bisporus; the AbChi1 gene is numbered AGABI2DRAFT_211936, and the AbCbm1 gene is numbered AGABI2DRAFT_193505. Co-silencing AbChi1 gene and AbCbm1 gene can promote the growth of Agaricus bisporus hyphae, thereby shortening the production cycle of Agaricus bisporus. In a specific embodiment, the Agaricus bisporus includes Agaricus bisporus A 15 In a specific embodiment, the reagent used for co-silencing the AbChi1 gene and the AbCbm1 gene includes an AbChi1 gene and AbCbm1 gene co-silencing vector.

[0034] The present invention also provides a vector for increasing the growth rate of Agaricus bisporus mycelium, the vector being a co-silencing vector of AbChi1 gene and AbCbm1 gene; the number of the AbChi1 gene is AGABI2DRAFT_211936, and the number of the AbCbm1 gene is AGABI2DRAFT_193505. In a specific embodiment, the vector is based on pCAMBIA1303 as a basic plasmid, contains a GPD promoter and an AbChi1 RNAi fragment expressed by the GPD promoter, and a TEF promoter and an AbCbM1 RNAi fragment expressed by the TEF promoter.

[0035] The present invention also provides a method for constructing a Agaricus bisporus strain with fast mycelium growth rate, comprising the following steps:

[0036] The vector described in the above technical solution is transformed into Agrobacterium to obtain Agrobacterium carrying the vector, and the Agrobacterium carrying the vector is used to infect Agaricus bisporus to obtain an Agaricus bisporus strain with fast mycelium growth rate. The present invention has no special limitation on the methods of transformation and Agrobacterium infection, and conventional methods can be used.

[0037] The present invention also provides an Agaricus bisporus strain with fast mycelium growth rate obtained by the construction method described in the above technical scheme.

[0038] To further illustrate the present invention, the preparation and application of a strain for accelerating the growth rate of Agaricus bisporus based on co-silencing of AbChi1 and AbCbm1 genes provided by the present invention are described in detail below in conjunction with the examples, but they should not be construed as limiting the scope of protection of the present invention.

[0039] Example 1

[0040] 1. Vector construction:

[0041] 1.AbChi1 RNAi fragment amplification

[0042] According to the RNAi target sequence selection principle, a fragment was selected from the AbChi1 gene ORF sequence and a specific primer sequence was designed. AbChi1 RNAi fragment amplification used AbChi1 cDNA as a template. The primer sequence, reaction system and PCR amplification procedure are shown in Table 1, Table 2 and Table 3:

[0043] Table 1 Primer sequences

[0044]

[0045] Table 2 Reaction system

[0046]

[0047] Table 3 PCR amplification program

[0048]

[0049] The PCR product was subjected to 1% agarose gel electrophoresis to obtain a single band. The gene fragment corresponding to this band was consistent with the size of the target fragment expected to be amplified. The electrophoresis results are shown in Figure 1 As shown, lane 1: AbChi1 RNAi fragment amplification product; lane 2: Trans2KplusII DNA marker.

[0050] The target gene band was precisely cut off under UV light and placed in a 2 ml sterile enzyme-free centrifuge tube, and DNA was recovered from agarose gel using the OMEGAD2000-1 PCR product recovery kit. AbChi1 RNAi fragments were amplified and recovered successfully.

[0051] 2. Linearization of pCambia1303-FvGPDA-AbCbM1-AbTEF plasmid

[0052] Eco91I digested the pcambia1303-FvGPDA-AbCbM1-AbTEF vector backbone (such as Figure 2 As shown, the vector backbone is based on pCAMBIA1303 as the basic plasmid, and the RNAi sequences of FvGPDA and AbCbM1 and the AbTEF fragment are inserted by conventional methods to construct the plasmid. The synthesis of this plasmid was entrusted to Wuhan Ruizhi Magic Cube Biotechnology Co., Ltd.), and the enzyme cutting system is shown in Table 4.

[0053] Table 4 Enzyme digestion system

[0054] name Addition amount Endonuclease Eco91I 1ul Buffer(10x) 2ul pcambia1303-FvGPDA-AbCbM1-AbTEF vector backbone 1ug <![CDATA[ddH2O]]> Make up to 20ul

[0055] The enzyme digestion reaction was carried out at 37℃ for 1h and inactivated at 80℃ for 10min.

[0056] The recovered PCR product was recombined and ligated with the pcambia1303-FvGPDA-AbCbM1-AbTEF vector backbone recovered after enzyme digestion. The recombination system is shown in Table 5.

[0057] Table 5 Recombination system

[0058] name Volume(ul) AbChi1 RNAi amplified fragment 0.5 pcambia1303-FvGPDA-AbCbM1-AbTEF linearized vector 1 5×CEIIbuffer 2 ExnaseII 1 <![CDATA[ddH2O]]> Make up to 10

[0059] After reacting at 37°C for 30 min, the mixture was quickly cooled on ice to obtain a co-silencing plasmid.

[0060] Transformation of recombinant products:

[0061] 1) Thaw TranT1 competent cells on ice.

[0062] 2) Take 10ul of the recombinant product (recombinant system mixture) and add it to 100ul of competent cells. Mix by gently tapping the tube wall (do not oscillate to mix), and let it stand on ice for 30min.

[0063] 3) After heat shock in a 42°C water bath for 45 seconds, immediately cool on ice for 2 to 3 minutes.

[0064] 4) Add LB medium (without antibiotics) and incubate at 37°C for 1 h (speed 200-250 rpm).

[0065] 5) Preheat the LB (kanamycin 50ug / ml) solid culture medium plates of the corresponding resistance in a 37°C incubator.

[0066] 6) Centrifuge at 5000 rpm for 5 min, discard 900 ul of supernatant, resuspend the cells with the remaining culture medium, and spread evenly on a plate containing 50 ug / ml kanamycin using a sterile spreader.

[0067] 7) Incubate the cells upside down in a 37°C incubator for 12 to 16 hours.

[0068] 3. Identification of recombinant products:

[0069] Several clones on the recombination reaction transformation plate were selected for colony PCR identification. The primers, amplification system and reaction conditions are shown in Table 6, Table 7 and Table 8.

[0070] Table 6 PCR identification primers

[0071]

[0072] Table 7 Amplification system

[0073] name Volume(ul) 2×TaqPlusMasterMixⅡ(DyePlus) 10 Primer AbCbm1F 1 Primer AbChi1-RNAiR 1 Bacterial liquid 1 <![CDATA[ddH2O]]> 7

[0074] Table 8 Reaction conditions

[0075]

[0076] The electrophoresis results are as follows Figure 3 As shown, according to Figure 3 It can be seen that all lanes have obvious bands around 700bp, indicating that they all carry the AbCbm1-AbChi1 RNAi fragment that silences two genes. After the colony in lane 1 was selected for expansion and culture, the plasmid was extracted for sequencing. The sequencing results were compared and the sequence was found to be correct, indicating that pCAMBIA1303-AbCbm1-AbChi1-RNAi was successfully constructed.

[0077] Figure 4 shown.

[0078] 2. Agrobacterium Transformation

[0079] 1. Transform Agrobacterium with pcambia1303-FvGPDA-AbCbm1-AbChi1-AbTEF vector

[0080] 1) Take the LBA4404 competent cell stored at -80℃ and wait for it to partially melt at room temperature. When it is in an ice-water mixture state, insert it into ice.

[0081] 2) Add 0.1 μg of plasmid DNA (pcambia1303-FvGPDA-AbCbm1-AbChi1-AbTEF (for Agaricus bisporus)) to 100 μl competent medium, stir the bottom of the tube by hand to mix, and place on ice for 5 min, in liquid nitrogen for 5 min, in a 37°C water bath for 5 min, and in an ice bath for 5 min.

[0082] 3) Add 700 μl of LB liquid culture medium without antibiotics and culture at 28°C with shaking for 2 to 3 hours.

[0083] 4) Collect the bacteria by centrifugation at 6000 rpm for 1 min, take about 100 μl of the supernatant, gently blow to resuspend the bacteria, and spread it on the LB plate containing the corresponding antibiotics (50 μg / ml kanamycin, 20 μg / ml rifampicin), and invert it in a 28°C incubator for 2 to 3 days.

[0084] 5) Pick the positive clones and transfer them to 10 ml LB medium (50 μg / ml kanamycin, 20 μg / ml rifampicin) and culture at 28°C with shaking until OD 600 When the pH is about 0.8, collect the cells by centrifugation, resuspend them in 10 ml PDB (As 20 mg / L) and set aside.

[0085] 2. Agrobacterium infection of Agaricus bisporus

[0086] 1) The Agaricus bisporus A15 strain was punched and inoculated into a potato dextrose agar medium (PDA), and cultured in a 25° C. incubator until the mycelium covered the entire plate (multiple point inoculation reduced the culture time).

[0087] 2) Punch a hole to obtain fresh mycelium, and add the mycelium to the Agrobacterium culture liquid carrying the pcambia1303-FvGPDA-AbCbM1-AbChi1-AbTEF plasmid prepared in the previous step, and co-cultivate at 25°C for 48 hours.

[0088] 3) After co-cultivation, the mycelium block was dried on sterile filter paper to remove excess Agrobacterium and then spread onto a PDA plate containing 40ug / ml hygromycin B and 200ug / ml cephalosporin. The plate was cultured in a 25°C incubator until mycelium began to extend significantly and resistant colonies were obtained, which lasted about 2 to 3 weeks.

[0089] 4) The resistant colonies obtained in step 3) were transferred to fresh PDA plates containing 40ug / ml hygromycin B and 200ug / ml cephalosporin and cultured at 25°C.

[0090] 3. T-DNA integration verification

[0091] 1) Take part of the resistant transformant mycelium obtained in step 4), grind it with liquid nitrogen, and extract genomic DNA according to the operating instructions of the tissue genomic DNA small-scale extraction kit.

[0092] 2) Primer design and synthesis.

[0093] The primers are AbCbm1F and AbChi1-RNAiR, and the primer sequences are shown in Table 6.

[0094] 3) PCR amplification

[0095] The amplification system and reaction conditions are shown in Table 9 and Table 10 respectively:

[0096] Table 9 Amplification system

[0097] name Volume(ul) 2×TaqPlusMasterMixⅡ(DyePlus) 10 Primer AbCbm1F (10uM) 1 Primer AbChi1-RNAiR (10uM) 1 Genomic DNA 1 <![CDATA[ddH2O]]> 7

[0098] Table 10 Reaction conditions

[0099]

[0100] The electrophoresis results are as follows Figure 5 As shown, lane 1: trans 2KplusII DNA marker; lane 2: amplification product of the wild-type control of Agaricus bisporus; lanes 3 to 5: amplification products of the pcambia1303-FvGPDA-AbCbM1-AbChi1-AbTEF transformant. The electrophoresis results showed that the plasmid was successfully transformed into Agaricus bisporus.

[0101] It was detected that pcambia1303-FvGPDA-AbCbM1-AbChi1-AbTEF transformants 1 to 3 were positive integration strains.

[0102] PCR experiments were performed to detect the expression levels of the gene AbCbm1 and the gene AbChi1 in the above three positive integration strains. The results showed that the silencing effect of co-silencing transformants 1 and 2 was very good, and the silencing effect of co-silencing transformant 3 was relatively weak. The present invention selects positive integration strains 1 and 2 for subsequent mycelium culture experiments.

[0103] The present invention utilizes the Agrobacterium transformation method to infect the Agaricus bisporus A15 strain, thereby realizing the construction of the AbChi1-AbCbm1 gene co-silencing strain.

[0104] After obtaining the positive integrated strains 1 and 2, the present invention inoculated the wild strain, the co-silenced strain and the AbChi1-RNAi strain (the construction method is the same as the co-silenced strain, only AbChi1 is silenced) and the AbCbm1-RNAi strain (the construction method is the same as the co-silenced strain, only AbCbm1 is silenced) cultured in the same period into the PDA culture medium using an Oxford cup (about 6 mm in diameter) of the same specifications and cultured for 14 days. Starting from the 6th day, photos of the mycelium growth on the plate were taken every two days to observe the mycelium growth status and record the mycelium growth rate.

[0105] Conclusion: According to Figure 6 It can be seen that the growth rate of the co-silenced strain is significantly faster than that of the AbChi1-RNAi strain and the AbCbm1-RNAi strain, and the speed is higher than that of the wild strain; the hyphae grow sparsely and irregularly when silenced alone, while the hyphae of the co-silenced strain are dense and healthy. The growth state of the co-silenced strain is significantly better than that of the wild strain, the AbChi1-RNAi strain, and the AbCbm1-RNAi strain.

[0106] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Application of co-silencing AbChi1 gene and AbCbm1 gene in improving the growth rate of Agaricus bisporus mycelium; the AbChi1 gene is numbered AGABI2DRAFT_211936, and the AbCbm1 gene is numbered AGABI2DRAFT_193505.

2. Application of co-silencing AbChi1 gene and AbCbm1 gene in improving the yield of Agaricus bisporus; the AbChi1 gene is numbered AGABI2DRAFT_211936, and the AbCbm1 gene is numbered AGABI2DRAFT_193505.

3. Application of co-silencing AbChi1 gene and AbCbm1 gene in preparing Agaricus bisporus strain with fast mycelium growth rate; the AbChi1 gene is numbered AGABI2DRAFT_211936, and the AbCbm1 gene is numbered AGABI2DRAFT_193505.

4. Application of co-silencing AbChi1 gene and AbCbm1 gene in shortening the production cycle of Agaricus bisporus; the AbChi1 gene is numbered AGABI2DRAFT_211936, and the AbCbm1 gene is numbered AGABI2DRAFT_193505.

5. The use according to any one of claims 1 to 4, characterized in that: The Agaricus bisporus includes Agaricus bisporus A 15 .

6. The use according to any one of claims 1 to 4, characterized in that: The reagent used for co-silencing the AbChi1 gene and the AbCbm1 gene includes an AbChi1 gene and an AbCbm1 gene co-silencing vector.

7. A carrier for increasing the growth rate of Agaricus bisporus mycelium, characterized in that: The vector is a co-silencing vector of the AbChi1 gene and the AbCbm1 gene; the number of the AbChi1 gene is AGABI2DRAFT_211936, and the number of the AbCbm1 gene is AGABI2DRAFT_193505.

8. The carrier according to claim 7, characterized in that The vector is based on pCAMBIA1303 as a basic plasmid, containing a GPD promoter and an AbChi1 RNAi fragment driven by the GPD promoter to express, and a TEF promoter and an AbCbM1 RNAi fragment driven by the TEF promoter to express.

9. A method for constructing a Agaricus bisporus strain with fast mycelium growth rate, characterized in that: The following steps are involved: The vector according to claim 7 or 8 is transformed into Agrobacterium to obtain Agrobacterium carrying the vector, and the Agrobacterium carrying the vector is used to infect Agaricus bisporus to obtain an Agaricus bisporus strain with fast mycelium growth rate.

10. The Agaricus bisporus strain with fast mycelial growth rate obtained by the construction method of claim 9.

Images