Application of rhizopus intro-rhizosporium in peanut cultivation

Abstract

The invention provides application of rhizopus intro-rhizosporium in peanut cultivation, and relates to the technical field of microorganisms. According to the application disclosed by the invention, it is proved for the first time that the growth process of peanuts can be promoted by the rhizopus intricadices BGSC-107, and the rhizopus intricadices BGSC-107 comprises the steps of increasing the germination rate of the peanuts, promoting the development of peanut rhizomes, increasing the weight of fresh flowers and the like. In addition, the application also proves that the rhizopus intro-rhizosporium BGSC-107 has an inhibition effect on the pathogenic bacterium Cercospora personata of the peanut black spot, and the occurrence of the peanut black spot can be effectively prevented and treated. The application also proves that the rhizopus intro-rhizosporium BGSC-107 has an inhibition effect on the pathogenic bacterium Cercospora personata. The invention further provides a preparation method of the microbial inoculum containing the rhizopus intro-rhizosporium BGSC-107, and a theoretical basis is provided for industrial production of a product containing the microbial inoculum.

Description

Technical Field

[0001] The present application relates to the field of microbial technology, and in particular to the application of Rhizospora endoradicata in peanut cultivation. Background Art

[0002] Rhizophagus intraradices is an arbuscular mycorrhizal fungus whose hyphae penetrate the epidermal cells of plant roots, so it can form a symbiotic system with 80% of higher plants on land. This symbiotic relationship can provide the mineral nutrients needed by plants, especially phosphorus and nitrogen, while plants provide photosynthetic products as a carbon source for fungi. Rhizophagus intraradices can form a symbiotic relationship with a variety of plants, such as soybeans, peppers, etc. However, there are no reports on the interaction between Rhizophagus intraradices and peanuts.

[0003] Peanut, also known as groundnut, is an annual herbaceous plant belonging to the genus Arachis of the Fabaceae family. It is native to South America. There are more than 100 countries in the world that cultivate peanuts, with Asia being the most common, followed by Africa. According to Chinese literature on peanuts, the history of peanut cultivation is about 100 years earlier than that of Europe. Peanuts are known as "vegetable meat" and have an oil content of up to 50%, excellent quality, and a fragrant smell. In addition to being edible, they are also used in printing and dyeing, papermaking, etc. Peanuts are also a traditional Chinese medicine, suitable for malnutrition, spleen and stomach disorders, coughing, sputum, and lack of milk. However, peanut cultivation is often plagued by a variety of diseases, resulting in reduced yields, and it is urgent to develop drugs and fertilizers that can effectively prevent and control peanut diseases. Summary of the invention

[0004] The present invention firstly finds that Rhizosporon endoradiata BGSC-107 can effectively promote the growth of peanuts and inhibit the growth of peanut pathogens, thereby playing a role in preventing and controlling peanut diseases, and provides a new biofertilizer for peanuts.

[0005] On the one hand, the present application provides the use of Rhizonhagus intraradices in promoting peanut growth and preventing and controlling peanut diseases, wherein the Rhizonhagus intraradices is Rhizonhagus intraradices BGSC-107.

[0006] Preferably, promoting the growth of peanuts includes increasing the germination rate of peanuts, promoting the development of peanut rhizomes, and increasing the fresh weight of peanuts.

[0007] Furthermore, the peanut diseases include peanut white rot and / or peanut black spot.

[0008] Preferably, the peanut disease is peanut black spot disease.

[0009] The control is achieved by inhibiting the pathogenic bacteria Cercospora personata of peanut black spot disease and Sclerotium rolfsii of peanut white rot disease.

[0010] Among them, Rhizospora endoradiata BGSC-107 can be cultured using a general fungal culture method.

[0011] On the other hand, the present application also provides a method for preparing a bacterial agent, wherein the bacterial agent includes Rhizocystis endoradicata BGSC-107.

[0012] Furthermore, the method comprises the following steps:

[0013] Step 1: Sterilize the substrate by mixing, wherein the substrate comprises peat and vermiculite in a mass ratio of (4-6):1;

[0014] Step 2, inoculating the root cyst BGSC-107 in a matrix at an inoculation rate of 1%-5% to obtain a bacterial matrix;

[0015] Step 3: sow alfalfa on the bacterial substrate, culture it for 0.5-1.5 months after germination, take the alfalfa roots, dry them, crush them into granules and obtain the bacterial agent.

[0016] Preferably, the matrix comprises peat and vermiculite in a mass ratio of 5:1.

[0017] Preferably, the inoculation amount is 3%.

[0018] Preferably, the culturing is performed for 1 month after germination.

[0019] In a preferred embodiment, the preparation method of the bacterial agent comprises the following steps:

[0020] Step 1: Mix peat and vermiculite in a mass ratio of (4-6):1, sterilize at 121°C for 1-2h to obtain a matrix;

[0021] Step 2, inoculating the root cyst BGSC-107 in a matrix at an inoculation amount of 1%-5% to obtain a bacterial matrix;

[0022] Step 3: sow alfalfa on the bacterial substrate, cultivate the alfalfa using conventional methods, and after germination, cultivate for 0.5-1.5 months, take the alfalfa roots, dry them at 35°C-45°C, and crush and granulate them to obtain the bacterial agent.

[0023] Those skilled in the art may select a specific drying method, crushing and granulation size according to actual needs, and no further limitation is made here.

[0024] On the other hand, the present application also provides a bacterial agent prepared by the method.

[0025] Preferably, the bacterial agent may include:

[0026] A1) Rhizocystis endoradiata BGSC-107;

[0027] A2) dead fungus suspension of Rhizospora endocystis BGSC-107;

[0028] A3) metabolites of Rhizocystis endoradicata BGSC-107;

[0029] A4) Rhizocystis endocystis BGSC-107 extract;

[0030] A5) mycelium of Rhizospora endocystis BGSC-107;

[0031] A6) Spores of Rhizospora endothecis BGSC-107.

[0032] Preferably, the bacterial agent can promote the growth of peanuts and / or prevent and treat peanut diseases.

[0033] On the other hand, the present application also provides a fertilizer comprising Rhizospora endocystis BGSC-107 or the bacterial agent as described.

[0034] Preferably, the fertilizer may include auxiliary materials, which may be appropriate solvents, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, thickeners, inclusion agents, humectants, flocculants and deflocculating agents, filter aids, release retardants, etc.

[0035] Preferably, the fertilizer is a peanut fertilizer, which can promote peanut growth and / or prevent and treat peanut diseases.

[0036] On the other hand, the present application also provides the use of the bacterial agent or the fertilizer in promoting peanut growth and preventing and controlling peanut diseases.

[0037] Furthermore, the peanut diseases include peanut white rot and / or peanut black spot.

[0038] Preferably, the peanut disease is peanut black spot disease.

[0039] It will be appreciated by those skilled in the art that the dosage of the bacterial agent or fertilizer can be adjusted according to actual conditions. In a preferred embodiment, the dosage of the bacterial agent or fertilizer is 2 g / plant.

[0040] The present invention has the following beneficial effects:

[0041] 1. This application first proves that Rhizospora endothelii BGSC-107 can promote the growth process of peanuts, including increasing the germination rate of peanuts, promoting the development of peanut rhizomes, and increasing the fresh weight of peanuts. In addition, this application also proves that Rhizospora endothelii BGSC-107 has an inhibitory effect on the pathogenic bacteria Cercospora globosum of peanut black spot disease, and can effectively prevent and control the occurrence of peanut black spot disease.

[0042] 2. This application also provides a method for preparing a bacterial agent containing Rhizospora endocystis BGSC-107, which provides a theoretical basis for the industrial production of products containing the bacterial agent. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] The drawings described herein are used to provide a further understanding of the present application, constitute a part of the present application, are used to explain the present application, and do not constitute an improper limitation on the present application. In the drawings:

[0044] Figure 1 This is a schematic diagram of peanut culture in Example 2. From left to right, pots 1-3 are blank control groups, and pots 4-6 are BGSC-107 treated groups. DETAILED DESCRIPTION

[0045] In order to more clearly explain the overall concept of the present application, the following is described in detail in conjunction with the accompanying drawings of the specification by way of embodiment. In the following description, a large number of specific details are provided to provide a more thorough understanding of the present invention. However, it is obvious to those skilled in the art that the present invention can be implemented without one or more of these details. In other examples, in order to avoid confusion with the present invention, some technical features well known in the art are not described.

[0046] If no specific conditions are specified in the examples, the experiments were carried out according to conventional conditions or conditions recommended by the manufacturer.

[0047] Among them, Rhizonhagus intraradices BGSC-107 was purchased from the Subtropical Arbuscular Mycorrhizal (AM) Fungi Resource Collection Center; the peanut variety was Qihua No. 1.

[0048] Unless otherwise specified, in the following embodiments, the reagents or instruments used without indicating the manufacturer are all conventional products that can be purchased commercially.

[0049] Unless otherwise stated, the experimental methods, detection methods, and preparation methods disclosed in the present invention all adopt conventional techniques in microbiology, biochemistry, analytical chemistry, cell culture, and related fields conventional in the technical field.

[0050] In addition, the "water" mentioned in the present invention includes any feasible water that can be used in the art, such as deionized water, distilled water, ion exchange water, double distilled water, high-purity water, and purified water.

[0051] In the following examples, unless otherwise specified, % means wt %, i.e. weight percentage.

[0052] Example 1 Cultivation of Rhizospora intraradicata

[0053] In this embodiment, alfalfa is used to ferment the root cyst BGSC-107, which specifically includes the following steps: sterilize the substrate at 121°C for 1h, put it into a flower pot (10cmx10cmx10cm), add the root cyst BGSC-107 at 2 / 3 of the substrate of the flower pot, and sow alfalfa at about 2cm above the substrate (the alfalfa seeds are sterilized by soaking in 10% sodium hypochlorite solution for 5min). Sow 30 seeds per pot, thin out the seedlings to 15 plants / pot one week after germination, water the Hogland nutrient solution twice a week, balance the water once a week, and finally remove the stem and leaf parts of the plant after germination for 1 month, dry the alfalfa roots, the root cyst BGSC-107 and the substrate at 35°C-45°C at low temperature, crush and granulate, and obtain the root cyst BGSC-107 bacterial agent. The blood cell counting method was used to detect the (live) spore content in the bacterial agent. The test results and specific conditions are shown in Table 1.

[0054] Table 1

[0055]

[0056]

[0057] Note: The ratio of matrix components is by mass.

[0058] From the results in Table 1, it can be seen that when peat and vermiculite with a mass ratio of 5:1 are used as the matrix material and the inoculation amount is 3%, the root cyst BGSC-107 inoculum with high spore content can be efficiently cultured within 1 month.

[0059] Example 2 Study on the effect of Rhizocystis endothelii on peanut growth promotion

[0060] In this example, the bacterial agent prepared by the optimized method of Example 1 was applied to the rhizosphere of peanuts to explore the promoting effect of Rhizosporium endothelialum on the growth of peanuts.

[0061] Specifically including: using nutrient soil as the peanut culture matrix, adding the root endocystis BGSC-107 bacterial agent obtained in Example 1 to 2 / 3 of the matrix of the flower pot (10cmx10cmx10cm), 2g / plant, and applying an equal amount of water to the blank control group. The peanut seeds were disinfected and sterilized by soaking in 75% alcohol for 1 minute and then in 10% sodium hypochlorite solution for 15 minutes, and then inoculated into the flower pots, sowing one seed in each pot, for a total of 60 pots. The germination rate of peanuts was counted after emergence. Watering with Hogland nutrient solution twice a week, and each pot was watered with the same amount of nutrient solution. The flower pots were arranged in random blocks, and the order was changed every week. After culturing for 21 days, the peanut plants were taken out, and the root length, plant height, and fresh weight of the peanuts were counted, and the trypan blue staining method was used for detection. The results and specific conditions are shown in Tables 2 and Figure 1 All data shown are averages.

[0062] Table 2

[0063] Group Germination rate (%) Root length (cm) Plant height (cm) Fresh weight(g) Mycorrhizal infection rate (%) BGSC-107 treatment group 94.25 11.59 15.41 5.60 75.26 Control group 73.76 7.23 6.57 2.59 -

[0064] From Table 2 and Figure 1 The results showed that Rhizospora intraradicata could effectively infect the rhizosphere of peanut and promote the germination and growth of peanut.

[0065] Example 3 Study on the effect of Rhizocystis endothelii on the prevention and treatment of common peanut diseases

[0066] To further explore the interaction between Rhizospora intraradicata and peanut, the preventive and therapeutic effects of Rhizospora intraradicata on common peanut diseases were tested in this example.

[0067] The plate confrontation method was used to inoculate equal amounts of root cysts of Rhizosporium endophyticum and the pathogen of peanut white rot disease, Sclerotium rolfsii (purchased from Testo Biological, No.: TS330731) on the PDA solid plate at the same time, and cultured at 22°C-28°C for one week to observe whether there was an inhibition zone between the two. The same method was used to test the antagonism of root cysts of Rhizosporium endophyticum with the pathogen of peanut root rot, Fusarium sp. (purchased from Testo Biological, No.: TS332341) and the pathogen of peanut black spot, Cercospora personata (purchased from Testo Biological, No.: TS332354), and the specific results are shown in Table 3.

[0068] Table 3

[0069] Antagonistic situation Sclerotium truncatum + Peanut root rot Fusarium - Cercospora globosa ++

[0070] Note: “++” indicates obvious inhibition band, “+” indicates inhibition band, and “-” indicates no inhibition band.

[0071] From the results in Table 3, it can be seen that there is an obvious inhibition zone between Rhizospora endothelii and the pathogenic bacteria of peanut black spot disease, Cercosporangium globosum, which proves that Rhizospora endothelii is antagonistic to the pathogenic bacteria of peanut black spot disease, and it has the potential to prevent and control peanut black spot disease.

[0072] Furthermore, a potted experiment for preventing and controlling peanut black spot was carried out in this embodiment. Specifically, it includes: using nutrient soil as the peanut culture matrix, adding the root root cyst BGSC-107 bacterial agent optimized in Example 1 to 2 / 3 of the matrix of the flower pot (10cmx10cmx10cm) at a dosage of 2g / plant, and applying an equal amount of water to the blank control group. Peanut seeds are sterilized by soaking in 75% alcohol for 1min and then in 10% sodium hypochlorite solution for 15min, and then inoculated into the flower pot, and 1 seed is sown per pot. Hogland nutrient solution is poured twice a week, and each pot is kept at the same amount of nutrient solution. The flower pots are arranged in random blocks and the order is changed every week. When the culture is 14 days, the pathogenic bacteria of peanut black spot are inoculated with Cercosporium globosum, and each peanut is inoculated with 250μL. When the culture is 21 days, the prevention and control situation is statistically analyzed, and the disease index and prevention and control effect are calculated with reference to Table 4. The prevention and control effect is shown in Table 5, and the disease index and prevention and control effect calculation method are as follows.

[0073]

[0074] Table 4

[0075] Disease level Grading Standards Level 0 No disease spots Level 1 The diameter of the lesion is 0-0.5mm Level 2 The diameter of the lesion is 0.5-2.0mm Level 3 The diameter of the lesion is 2.0-5.0 mm Level 4 The diameter of the lesion is greater than 5.0 mm

[0076] Table 5

[0077] Treatment Group Control effect / % BGSC-107 treatment group 88.60 Blank control /

[0078] From the results in Table 5, it can be seen that the root cyst BGSC-107 fungicide can effectively prevent and control peanut black spot disease.

[0079] The above is only an embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various changes and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the scope of the claims of the present application.

Claims

1. Application of Rhizonhagus intraradices in promoting peanut growth and preventing and controlling peanut diseases, characterized in that: The Rhizospora endoradicata is Rhizospora endoradicata BGSC-107, and the peanut diseases include peanut white rot and / or peanut black spot.

2. A method for preparing a bacterial agent, characterized in that: The method comprises the following steps: Step 1: Sterilize the substrate by mixing, wherein the substrate comprises peat and vermiculite in a mass ratio of (4-6):1; Step 2, inoculating the root cyst BGSC-107 in a matrix at an inoculation rate of 1%-5% to obtain a bacterial matrix; Step 3: sow alfalfa on the bacterial substrate, culture it for 0.5-1.5 months after germination, take the alfalfa roots, dry them, crush them and granulate them to obtain the bacterial agent.

3. The bacterial agent prepared according to the method of claim 2.

4. A fertilizer, characterized in that: Contains the bacterial agent as claimed in claim 3.

5. The use of the microbial agent as claimed in claim 3 or the fertilizer as claimed in claim 4 in promoting peanut growth and preventing and controlling peanut diseases, characterized in that: The peanut diseases include peanut white rot and / or peanut black spot.

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