Microecological preparation for improving disease resistance of medlar and application thereof
The microecological preparation made of micrococci gazella and Clostridium Senegal combined with astaxanthin solves the serious disease problems in wolfberry cultivation, significantly improves the disease resistance and yield of wolfberry, and has excellent synergistic effect.
Patent Information
- Application Number
- CN202411316673.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-20
- Publication Date
- 2025-02-14
AI Technical Summary
During wolfberry planting, the number of soil pathogens increases due to repeated crops, resulting in an increase in the incidence of wolfberry root rot and leaf spot diseases, which seriously affects the economic income of farmers.
Micrococci Garcinia and Clostridium Senegal were used to make microecological preparations, and the anti-disease of wolfberry was significantly improved by watering and foliar spraying.
It significantly reduces the incidence of leaf spot disease and root rot of wolfberry, improves the disease resistance of wolfberry, promotes the growth of wolfberry, and increases yield, and has better results than ordinary microecological preparations on the market.
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Abstract
Description
Technical Field
[0001] The invention relates to the field of wolfberry planting, in particular to a microecological preparation for improving the disease resistance of wolfberry and a use thereof. Background Art
[0002] Ecological agriculture, as an environmentally friendly, resource-saving and high-efficiency agricultural model, has received widespread attention. As an important part of ecological agriculture, plant microecological preparations refer to microbial preparations that promote plant growth and health through the symbiotic relationship between specific microbial strains and plants. They have many advantages such as promoting plant growth, increasing yield and quality, and improving soil environment, and are gradually being widely used in modern ecological agriculture. These microorganisms include bacteria, fungi, actinomycetes, etc., which can improve plant stress tolerance and growth performance through disease resistance and growth-promoting properties. Recent studies have shown that plant microecological systems play an important role in plant growth, health and adaptation to the environment. Not only that, changes in microbial communities are also believed to be related to the ability of plants to resist diseases.
[0003] Lycium barbarum is a perennial deciduous shrub. It is cultivated artificially and tied to a rack and pruned into a small tree. It is a plant with multiple economic traits and exhibits better stress resistance than other plants. The fruit is a precious Chinese medicinal material. It contains polysaccharides, multiple vitamins, amino acids and glycosides. Its main functions are to nourish the liver and kidney, nourish the liver, improve eyesight, promote blood circulation, and improve essence. In recent years, the area of wolfberry cultivation has continued to expand, and the continuous cropping of wolfberry has led to an increase in pathogens in the soil; the incidence of plant diseases such as wolfberry root rot in new and old production areas has increased year by year and has become increasingly serious. It has seriously affected the economic income of local wolfberry farmers and has a certain destructive effect on the entire industry chain. Therefore, it is urgent to develop a microecological preparation that has outstanding effects, can promote the growth of wolfberry, and improve the disease resistance of wolfberry. Summary of the invention
[0004] The purpose of the present invention is to provide a microecological preparation for improving the disease resistance of wolfberry and its use, so as to solve the problems existing in the above-mentioned prior art. The present invention combines astaxanthin with Micrococcus luteus and Clostridium senegal to prepare a microecological preparation, which exerts a synergistic effect and significantly improves the disease resistance of wolfberry.
[0005] To achieve the above object, the present invention provides the following solutions:
[0006] The present invention provides a bacterial composition for improving the disease resistance of wolfberry, characterized in that it comprises Micrococcus luteus and Clostridium senegalense;
[0007] The Micrococcus luteus has been deposited in the China Center for Type Culture Collection, the strain deposit number is CCTCC NO: M2021839, the deposit date is July 8, 2021, and the deposit address is Wuhan University, Wuhan, China;
[0008] The senegalese bacterium has been deposited in the China Center for Type Culture Collection, the strain deposit number is CCTCC NO: 2022707, the deposit date is May 24, 2022, and the deposit address is Wuhan University, Wuhan, China.
[0009] The present invention also provides the use of the bacterial composition in preparing a microecological preparation for improving the disease resistance of wolfberry.
[0010] The invention also provides a microecological preparation for improving the disease resistance of wolfberry, comprising the bacterial composition, astaxanthin and auxiliary materials.
[0011] Preferably, in the probiotic preparation, the ratio of the effective viable counts of the Micrococcus luteus to the Clostridium senegalense is 1:1; and the total effective viable counts of the Micrococcus luteus and the Clostridium senegalense is not less than 2×10 7 CFU / g.
[0012] Preferably, in the microecological preparation, the mass percentage of astaxanthin is 2%.
[0013] Preferably, in the microecological preparation, the mass percentage of the auxiliary material is 28%;
[0014] The auxiliary materials include beta-cyclodextrin, sodium dihydrogen phosphate, disodium hydrogen phosphate and sodium stearate; by weight, the auxiliary materials are composed of 16 parts of beta-cyclodextrin, 2 parts of sodium dihydrogen phosphate, 4 parts of disodium hydrogen phosphate and 6 parts of sodium stearate.
[0015] The present invention also provides a method for preparing the microecological preparation, which is characterized by comprising the following steps:
[0016] The Micrococcus luteus and the Clostridium senegal were activated and cultured to the logarithmic growth phase, and the cells were collected by centrifugation, washed, and diluted to 3×10 7 CFU / mL, and obtain a bacterial solution; the bacterial solution of Micrococcus luteus and the bacterial solution of Clostridium senegal are mixed evenly in a volume ratio of 1:1 to obtain a mixed bacterial solution;
[0017] By weight, 16 parts of β-cyclodextrin, 2 parts of sodium dihydrogen phosphate, 4 parts of disodium hydrogen phosphate and 6 parts of sodium stearate are mixed evenly to obtain an auxiliary material;
[0018] By weight, 70 parts of the mixed bacterial solution, 28 parts of the auxiliary materials and 2 parts of astaxanthin are evenly mixed to obtain the microecological preparation.
[0019] The present invention also provides the use of the bacterial composition or the microecological preparation in improving the disease resistance of wolfberry and / or promoting the growth of wolfberry.
[0020] The present invention also provides a method for improving the disease resistance of wolfberry and / or promoting the growth of wolfberry, comprising the steps of applying the microecological preparation in the growth environment of wolfberry before and after planting wolfberry.
[0021] Preferably, the application method includes irrigation and foliar spraying.
[0022] The present invention discloses the following technical effects:
[0023] The microecological preparation prepared by the present invention can significantly improve the disease resistance of wolfberry and promote the growth of wolfberry, and the effect is better than the common microecological preparations available on the market. The present invention prepares a microecological preparation by compounding Micrococcus luteus and Clostridium senegal and mixing them with astaxanthin, which has an excellent effect on improving the disease resistance of wolfberry. Compared with the microecological preparation prepared by using a single type of bacteria or the microecological preparation without astaxanthin, the two bacterial combinations and astaxanthin have obvious synergistic effects after being used in combination. The present invention provides a new microecological preparation for improving the disease resistance of wolfberry, and it has a wide source, simple preparation, and broad application prospects. DETAILED DESCRIPTION
[0024] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.
[0025] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.
[0026] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.
[0027] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.
[0028] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.
[0029] In the present invention, Micrococcus luteus with a preservation number of CCTCC NO: M 2021839 and Clostridium senegalense with a preservation number of CCTCC NO: 2022707 were purchased from China Center for Type Culture Collection. The public can purchase the above strains from the above collection institutions.
[0030] Example 1
[0031] Take the preparation of 1000 g of the microecological preparation of the present invention as an example.
[0032] Micrococcus luteus with a preservation number of CCTCC NO: M 2021839 and Clostridium senegalense with a preservation number of CCTCC NO: 2022707 were activated and expanded to the logarithmic growth phase, and the cells were collected by centrifugation, washed, and diluted to 3×10 7 CFU / mL, take 500mL of bacterial solution respectively, mix them evenly, and obtain 1000mL of mixed bacterial solution;
[0033] 160 g of β-cyclodextrin, 20 g of sodium dihydrogen phosphate, 40 g of disodium hydrogen phosphate and 60 g of sodium stearate were mixed to obtain 280 g of auxiliary material;
[0034] Take 700 g of the above mixed bacterial liquid, add 280 g of auxiliary materials and 20 g of astaxanthin, mix well, and obtain.
[0035] Comparative Example 1
[0036] The only difference from Example 1 is that 1000 mL of bacterial solution of Micrococcus luteus with a preservation number of CCTCC NO: M 2021839 is used to replace the above mixed bacterial solution.
[0037] Comparative Example 2
[0038] The only difference from Example 1 is that 1000 mL of bacterial solution of Clostridium senegalensis with a preservation number of CCTCC NO: 2022707 is used to replace the above mixed bacterial solution.
[0039] Comparative Example 3
[0040] The only difference from Example 1 is that 500 mL of uninoculated Micrococcus luteus culture medium and 500 mL of uninoculated Clostridium senegal culture medium are respectively taken, mixed evenly, and replaced with the above mixed bacterial solution.
[0041] Comparative Example 4
[0042] The only difference from Example 1 is that 20 g of auxiliary material made of 11.43 g of β-cyclodextrin, 1.43 g of sodium dihydrogen phosphate, 2.86 g of disodium hydrogen phosphate and 4.28 g of sodium stearate is used to replace the above 20 g of astaxanthin.
[0043] Comparative Example 5
[0044] The "Zhongnong Lvkang" brand anti-replanting microecological preparation (purchased from Zhongnong Lvkang (Beijing) Biotechnology Co., Ltd.) was used as an agent to improve the disease resistance of wolfberry and promote its growth.
[0045] Effect verification example
[0046] 1. Wolfberry planting experiment
[0047] The 4-year-old 'Ningqi No. 7' was planted in sandy loam in Yinchuan City, with a cultivation pattern of 1.5m×2m plant spacing, an area of 18 mu, and sandy loam with normal fertility. The experiment started in March, and the forest was divided into 3 large areas, each of which was further divided into 6 small areas. The 6 small areas in each large area were treated with the microecological preparations prepared according to any one of Example 1 and Comparative Examples 1-5, and different large areas were used as biological replicates.
[0048] Before planting, dig a planting pit of 20cm in diameter, 35cm deep, with a pit spacing of 1.2 meters, and the corresponding microecological preparation prepared by any one of Example 1 and Comparative Examples 1-5 is irrigated in the planting pit, and the consumption is 10kg per planting pit. Before the arrival of the full flowering period of wolfberry (May 31), at a distance of 40cm from the wolfberry trunk, dig a deep 20cm watering hole, supplement the microecological preparation 0.5kg prepared by any one of Example 1 and Comparative Examples 1-5, irrigate once every other day, and irrigate 10 times in total, to June 18. At the same time, starting from June 1, the microecological preparation prepared by any one of Example 1 and Comparative Examples 1-5 is sprayed every 15 days, and it is advisable to evenly spray the whole plant leaf surface without dripping, and spray 3 times in total.
[0049] Except for the application of different microecological products, all fields adopt the same management measures, that is, wolfberry planting, fertilizer application, watering, pruning, pest and disease control and other management measures are carried out in accordance with wolfberry production standards.
[0050] 2. Wolfberry harvesting and statistics
[0051] 2.1 Leaf spot statistics
[0052] On July 6, 18 days after the supplementary irrigation, the incidence of wolfberry leaf spot disease was tested.
[0053] One long branch was taken from each wolfberry plant in the east, south, west and north, and the number of diseased leaves and the area of lesions were counted. The disease grade classification standard is: grade 0, no lesions; grade 1, lesions occupy 1%-10% of the leaf area; grade 2, lesions occupy 11%-25% of the leaf area; grade 3, lesions occupy 26%-50% of the leaf area; grade 4, lesions occupy more than 51% of the leaf area.
[0054] The calculation formulas for leaf morbidity and disease index are as follows:
[0055] The incidence of leaves = ∑ the number of diseased leaves / ∑ the total number of leaves × 100%;
[0056] Disease index = [∑(number of diseased leaves at each level × corresponding level value) / (total number of leaves surveyed × highest disease level)] × 100.
[0057] 2.2 Fruit picking and weighing
[0058] 25 days after the supplementary irrigation, wolfberry picking began, and the harvest was carried out once every 7 days, for a total of 5 times (i.e., from July 13 to August 14). After drying, the weight of dried wolfberry in each treatment area was counted to calculate the equivalent yield.
[0059] 2.3 Root rot statistics
[0060] After the fruits are harvested, the incidence rate is recorded per plant. According to the disease grading standards, the number of plants with root rot at each level is counted and the disease index is calculated. The specific grading standards are shown in Table 1.
[0061] Table 1 Lycium barbarum root rot disease classification standard
[0062]
[0063] The calculation formula is as follows:
[0064] Disease index = [Σ(number of diseased plants at each level × relative level value) / (total number of plants surveyed × highest level representative value)] × 100;
[0065] The incidence of root rot = number of diseased plants / total number of plants surveyed × 100%.
[0066] 3. Experimental results
[0067] The results are shown in Table 2. According to Table 2, the microecological agent prepared in Example 1 of the present invention can significantly reduce the incidence and disease index of leaf spot and root rot of wolfberry, improve the disease resistance of wolfberry, and significantly promote the growth of wolfberry, greatly increase the yield of wolfberry, and the effect is better than the general commercially available microecological preparation (Comparative Example 5).
[0068] Combined with the experimental results of Comparative Examples 1-4, it can be found that the bacterial composition selected by the present invention is compounded with astaxanthin, that is, the combination of Micrococcus luteus, Clostridium senegal and astaxanthin, has a particularly excellent effect on improving the disease resistance of wolfberry, and the effects of using only one bacterial liquid in combination with astaxanthin, using only astaxanthin or a mixed liquid without adding astaxanthin are significantly worse than those of the probiotic preparation, indicating that Micrococcus luteus, Clostridium senegal and astaxanthin have a synergistic effect.
[0069] Table 2 The incidence of wolfberry leaf spot and root rot and the statistics of dry fruit weight
[0070]
[0071]
[0072] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.
Claims
1. A bacterial composition for improving the disease resistance of wolfberry, characterized in that: These include Micrococcus luteus and Clostridium senegalense; The preservation number of the Micrococcus luteus is CCTCC NO: M 2021839; the preservation number of the Clostridium senegalense is CCTCC NO: 2022707.
2. Use of the bacterial composition as claimed in claim 1 in the preparation of a probiotic preparation for improving the disease resistance of wolfberry.
3. A microecological preparation for improving the disease resistance of wolfberry, characterized in that: The method comprises the bacterial composition as claimed in claim 1, astaxanthin and auxiliary materials.
4. The microecological preparation according to claim 3, characterized in that: In the probiotic preparation, the ratio of the effective viable counts of the Micrococcus luteus to the Clostridium senegalense is 1:1; the total effective viable counts of the Micrococcus luteus and the Clostridium senegalense is not less than 2×10 7 CFU / g.
5. The microecological preparation according to claim 3, characterized in that: In the microecological preparation, the mass percentage of astaxanthin is 2%.
6. The microecological preparation according to claim 3, characterized in that: In the microecological preparation, the mass percentage of the auxiliary material is 28%; The auxiliary materials include beta-cyclodextrin, sodium dihydrogen phosphate, disodium hydrogen phosphate and sodium stearate; by weight, the auxiliary materials are composed of 16 parts of beta-cyclodextrin, 2 parts of sodium dihydrogen phosphate, 4 parts of disodium hydrogen phosphate and 6 parts of sodium stearate.
7. The method for preparing the probiotic preparation according to any one of claims 3 to 6, characterized in that: The following steps are involved: The Micrococcus luteus and the Clostridium senegal were activated and cultured to the logarithmic growth phase, and the cells were collected by centrifugation, washed, and diluted to 3×10 7 CFU / mL, and obtain a bacterial solution; the bacterial solution of Micrococcus luteus and the bacterial solution of Clostridium senegal are mixed evenly in a volume ratio of 1:1 to obtain a mixed bacterial solution; By weight, 16 parts of β-cyclodextrin, 2 parts of sodium dihydrogen phosphate, 4 parts of disodium hydrogen phosphate and 6 parts of sodium stearate are mixed evenly to obtain an auxiliary material; By weight, 70 parts of the mixed bacterial solution, 28 parts of the auxiliary materials and 2 parts of the astaxanthin are evenly mixed to obtain the microecological preparation.
8. Use of the bacterial composition according to claim 1 or the probiotic preparation according to any one of claims 3 to 6 in improving the disease resistance of wolfberry and / or promoting the growth of wolfberry.
9. A method for improving disease resistance of wolfberry and / or promoting growth of wolfberry, characterized in that: The method comprises the steps of applying the probiotic preparation according to any one of claims 3 to 6 in the growth environment of wolfberries before and after planting wolfberries.
10. The method according to claim 9, characterized in that The application methods include irrigation and foliar spraying.