Preparation method of functional biological composition for promoting healthy growth and quality improvement of plants

Abstract

The invention provides a preparation method of a biological composition for functionally promoting healthy growth and quality improvement of plants, and belongs to the technical field of compound microbial fertilizers. The compound microbial fertilizer is prepared from the following raw materials in parts by weight: 0.5 to 2 parts of growth factors, 1 to 3 parts of bacillus subtilis, 1 part of bacillus cereus, 2 to 6 parts of trichoderma harzianum and 1 to 3 parts of trichoderma pseudokoningii. The compound microbial fertilizer disclosed by the invention can promote crop growth, improve product quality, and also can achieve the purposes of reducing the rotting speed after harvesting and prolonging the shelf life.

Description

Technical Field

[0001] This invention belongs to the field of compound microbial fertilizer technology, and in particular relates to a method for formulating a biological combination that functionally promotes healthy plant growth and improves plant quality. Background Technology

[0002] European pears (Pyrus communis L.) are one of the world's two major cultivated pear types, alongside Oriental pears. However, European pears require further ripening to reach their optimal state of being soft, juicy, and fragrant. The shelf life of European pears is generally 7-10 days, with the optimal eating period typically being 2-3 days. Therefore, accurately determining the optimal eating period is crucial. The pear is a type of European pear.

[0003] Ripe pears contain almost no stone cells, resulting in a creamy, soft, and delicate texture that requires minimal chewing. They can be eaten by scooping with a spoon or sucking through a straw. Their sweetness is highly concentrated, and their aroma is rich, often described as reminiscent of honey or cheese. Pears are believed to have lung-moistening, dryness-relieving, cough-suppressing, phlegm-reducing, blood-nourishing, and muscle-regenerating effects. As a climacteric fruit, pears continue to ripen after harvesting due to the presence of ethylene. Simply put, pears are a soft, creamy fruit that needs to be fully ripe before consumption. Pears have a typical gourd shape with a slender neck and a round body. Unripe pears are green, turning yellow with a reddish tinge when ripe. They require further ripening after harvesting, softening from firm to tender. These characteristics necessitate special measures and methods for improving the quality and preserving the freshness of pears.

[0004] Currently, there is limited research on pears. In 2007, Jia Xiaohui et al. used Abate pears as test material. To accelerate ripening, they sealed and fumigated the fruit at 20℃ for 24 hours using three methods: injecting 50 and 100 μL / L ethylene gas into a preservation bag, and adding bananas. They then measured the changes in various physiological indicators during the ripening process. The results showed that all three treatments shortened the ethylene catalytic initiation time to varying degrees, increased fruit respiration intensity, and accelerated the rate at which the peel color changed from green to yellow. Ethylene treatment significantly promoted the ripening process of Abate pears, and the degree of soft ripeness was basically consistent. Using 100 μL / L ethylene gas for ripening treatment could advance the fruit to the edible stage by 2-3 days. In 2016, Ma Fengli et al. aimed to clarify the physiological and biochemical changes during the ripening process of European pears and the correlation between various physiological indicators, to find characteristic indicators reflecting the ripening process, and to achieve rapid and non-destructive detection of the ripening degree of European pears. Physiological and biochemical indicators of the fruit at different ripening times were compared. The peel color changed from green to yellow, with a gradual increase in brightness; firmness, titratable acid content, and vitamin C content showed a gradual decreasing trend, while soluble solids content showed an initial increase followed by a decrease; ethylene and respiratory climacteric peaks appeared at 6 days of ripening. Additionally, the fruit's firmness decreased to 3 kg·cm². -2The optimal time for fruit consumption is during the ripening process. The optimal time for fruit consumption can be determined non-destructively by measuring the peel color or chlorophyll fluorescence parameters during the ripening process.

[0005] As a fruit enjoyed by people of all ages, there is currently almost no research on improving the quality and shelf life of pears. Only by ensuring superior quality and extending the shelf life can consumer demand be met; and producers can only avoid the pressure of concentrated market entry by extending the market supply cycle. Summary of the Invention

[0006] In view of this, one of the objectives of the present invention is to provide a compound microbial fertilizer.

[0007] The second objective of this invention is to provide a method for using the compound microbial fertilizer.

[0008] The third objective of this invention is to provide the application of the compound microbial fertilizer or the method of use in improving the quality of fruits and vegetables.

[0009] The fourth objective of this invention is to provide the application of the compound microbial fertilizer or the method of use in improving the shelf life of fruits and vegetables.

[0010] To achieve the above-mentioned objectives, the present invention provides the following technical solution: A compound microbial fertilizer, wherein the compound microbial fertilizer comprises the following raw materials by weight: 0.5-2 parts of growth factor, 1-3 parts of Bacillus subtilis, 1 part of Bacillus cereus, 2-6 parts of Trichoderma harzianum and 1-3 parts of Trichoderma koningii.

[0011] Preferably, the compound microbial fertilizer comprises the following raw materials by weight: 1-1.5 parts of growth factor, 2 parts of Bacillus subtilis, 1 part of Bacillus cereus, 3-4 parts of Trichoderma harzianum, and 2 parts of Trichoderma koningii.

[0012] Preferably, the viable counts of Bacillus subtilis, Bacillus cereus, Trichoderma harzianum, and Trichoderma konjac are all greater than 10. 8 cfu / g.

[0013] Preferably, the growth factor is any one or both of yam extract and chitosan.

[0014] The present invention also provides a method for using the compound microbial fertilizer, wherein the compound microbial fertilizer is mixed with water to obtain a compound microbial fertilizer solution; after the fruit tree flower buds sprout, the compound microbial fertilizer solution is applied twice with water; after fruiting, the compound microbial fertilizer solution is sprayed on the plants.

[0015] Preferably, the compound microbial fertilizer is mixed with water at a mass ratio of 1:300~500.

[0016] Preferably, after the fruit tree flower buds sprout, the application rate of compound microbial fertilizer solution is 5-7 L / mu.

[0017] The present invention also provides the application of the compound microbial fertilizer or the method of application in improving the quality of fruits and vegetables.

[0018] The present invention also provides the application of the compound microbial fertilizer or the method of application in improving the shelf life of fruits and vegetables.

[0019] Preferably, the fruit or vegetable includes pears.

[0020] Compared with the prior art, the present invention has the following beneficial effects: This invention provides a compound microbial fertilizer that utilizes the synergistic effect of Bacillus subtilis, Bacillus cereus, Trichoderma harzianum, and Trichoderma koningii to promote crop growth, improve product quality, and reduce post-harvest decay while extending shelf life.

[0021] Biological Preservation Instructions The Bacillus subtilis (Latin name) described in this invention Bacillus subtilis (), depositary institution: China General Microbiological Culture Collection Center, depositary address: No. 13, Zhongguancun North First Street, Haidian District, Beijing, depositary number: CGMCC NO.1442, depositary date: August 22, 2005. Attached Figure Description

[0022] Figure 1 The effects of different compound microbial fertilizers on the firmness of pear fruits. Detailed Implementation

[0023] This invention provides a compound microbial fertilizer, which, by weight, comprises the following raw materials: 0.5-2 parts of growth factor, 1-3 parts of Bacillus subtilis, 1 part of Bacillus cereus, 2-6 parts of Trichoderma harzianum, and 1-3 parts of Trichoderma koningii. Preferably, the compound microbial fertilizer, by weight, comprises the following raw materials: 1-1.5 parts of growth factor, 2 parts of Bacillus subtilis, 1 part of Bacillus cereus, 3-4 parts of Trichoderma harzianum, and 2 parts of Trichoderma koningii.

[0024] The compound microbial fertilizer of the present invention contains Bacillus subtilis, Bacillus cereus, Trichoderma harzianum, and Trichoderma kosneri, each with a viable count greater than 10. 8cfu / g. In this invention, the Bacillus subtilis strain has the biodeposit number CGMCC NO.1442, which was deposited on August 22, 2005, at the China General Microbiological Culture Collection Center (address: No. 13, Zhongguancun North First Street, Haidian District, Beijing), with the biodeposit number CGMCC NO.1442. The Trichoderma ts. is preferably the Trichoderma ts. disclosed in patent CN106801083A, with the biodeposit number CGMCC NO.1443. In a specific embodiment of this invention, Bacillus cereus was purchased from Xi'an Zhonghe Crop Science Co., Ltd., and Trichoderma harzianum was purchased from Shandong Changtai Biotechnology Co., Ltd.

[0025] In this invention, the growth factor is any one or both of yam extract and chitosan. The preparation method of the yam extract includes the following steps: Yam is cut into small pieces, dried at 85°C, pulverized, and sieved to obtain yam powder, preferably pulverized and sieved through an 80-mesh sieve. 70% ethanol (volume fraction) is used as the extraction solution, with a preferred material-to-liquid ratio of 1g:20mL; extraction is preferably performed at 50°C for 120 minutes. After filtration, the supernatant is collected, concentrated, and dried to obtain the yam extract. This invention does not limit the concentration and drying methods of the supernatant; conventional methods in the art are acceptable. When yam extract and chitosan are added simultaneously, they are mixed at a mass ratio of 1:1. In a specific embodiment of this invention, the chitosan was purchased from Shandong Suihua Biotechnology Co., Ltd.

[0026] The compound microbial fertilizer of this invention employs a synergistic strategy of "bacteria + fungi" and "growth promotion + disease prevention." They are not simply superimposed, but rather complement and synergistically enhance each other through different mechanisms of action, jointly constructing a rhizosphere microecological environment conducive to plant health. Different microorganisms have varying adaptability to the environment (temperature, humidity, pH). After compounding, under different soil conditions, they function like a coordinated army, displacing pathogens from different dimensions (site occupation vs. area occupation), better exerting synergistic effects and ensuring stable efficacy. Furthermore, different microorganisms produce different types of antibacterial substances, which, working together, can inhibit a wider range of pathogens (fungi and bacteria) and reduce the risk of pathogens developing drug resistance.

[0027] In the compound microbial fertilizer of this invention, the growth factor not only provides fuel for the rapid start-up and efficient operation of beneficial microorganisms, but also acts as a crop resistance-inducing factor, achieving a synergistic effect of 1+1>2 with the microbial agent.

[0028] The present invention also provides a method for using the compound microbial fertilizer, wherein the compound microbial fertilizer is mixed with water to obtain a compound microbial fertilizer solution; after the fruit tree flower buds sprout, the compound microbial fertilizer solution is applied twice with water; after fruiting, the compound microbial fertilizer solution is sprayed on the plants.

[0029] In the method of use of this invention, the compound microbial fertilizer and water are preferably mixed at a mass ratio of 1:300-500, more preferably at a mass ratio of 1:400. After the Spring Festival (when fruit tree flower buds sprout), with the first watering, the application rate of the compound microbial fertilizer solution is preferably 5-7 L / mu, more preferably 6 L / mu; preferably, it is applied twice with water, with an interval of 3 weeks between the two applications. Applying the compound microbial fertilizer of this invention after the fruit tree flower buds sprout allows the microorganisms in the fertilizer to quickly form a dominant bacterial community in the soil, improving the soil microenvironment in the rhizosphere and promoting healthy crop growth. After the fruit tree has sprouted, it is preferably sprayed once every two weeks, with the spraying time ensuring that the solution is evenly coated on the entire plant without dripping.

[0030] The present invention also provides the application of the compound microbial fertilizer or the method of application in improving the quality of fruits and vegetables and extending their shelf life, or both, wherein the fruits and vegetables include pears and berries.

[0031] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0032] Example 1 A compound microbial fertilizer, wherein the compound microbial fertilizer comprises the following raw materials by weight: 1 part growth factor, 2 parts Bacillus subtilis (CGMCC NO.1442), 1 part Bacillus cereus, 4 parts Trichoderma harzianum and 2 parts Trichoderma ts.

[0033] Among them, the viable counts of Bacillus subtilis, Bacillus cereus, Trichoderma harzianum, and Trichoderma konjac all exceeded 10. 8 cfu / g.

[0034] The growth factor is a mixture of yam extract and chitin in a 1:1 mass ratio.

[0035] The preparation method of yam extract is as follows: After cutting the yam into pieces, drying it at 85℃, and then pulverizing it through an 80-mesh sieve, yam powder was obtained. The yam powder was mixed with 70% ethanol at a ratio of 1g:20mL and extracted at 50℃ for 120min. After filtration, the supernatant was collected, concentrated, and dried to obtain yam extract.

[0036] Example 2 A compound microbial fertilizer, wherein the compound microbial fertilizer comprises the following raw materials by weight: 2 parts growth factor, 2 parts Bacillus subtilis (CGMCC NO.1442), 1 part Bacillus cereus, 2 parts Trichoderma harzianum and 1 part Trichoderma ts.

[0037] Among them, the viable counts of Bacillus subtilis, Bacillus cereus, Trichoderma harzianum, and Trichoderma konjac all exceeded 10. 8 cfu / g.

[0038] The growth factor is yam extract, and the preparation method of yam extract is the same as in Example 1.

[0039] Example 3 A compound microbial fertilizer, wherein the raw materials of the compound microbial fertilizer are as follows by weight: 1.5 parts of growth factor, 2 parts of Bacillus subtilis (CGMCC NO.1442), 1 part of Bacillus cereus, 6 parts of Trichoderma harzianum and 3 parts of Trichoderma ts.

[0040] Among them, the viable counts of Bacillus subtilis, Bacillus cereus, Trichoderma harzianum, and Trichoderma konjac all exceeded 10. 8 cfu / g.

[0041] Among them, the growth factor is chitosan.

[0042] Example 4 A compound microbial fertilizer, wherein the compound microbial fertilizer comprises the following raw materials by weight: 1 part growth factor, 3 parts Bacillus subtilis (CGMCC NO.1442), 1 part Bacillus cereus, 4 parts Trichoderma harzianum and 2 parts Trichoderma ts.

[0043] Among them, the viable counts of Bacillus subtilis, Bacillus cereus, Trichoderma harzianum, and Trichoderma konjac all exceeded 10. 8 cfu / g.

[0044] The growth factor is a mixture of yam extract and chitin in a mass ratio of 1:1. The preparation method of the yam extract is the same as in Example 1.

[0045] Comparative Example 1 The difference from Example 1 is that Bacillus subtilis (CGMCC NO.1442) and Bacillus cereus were not added.

[0046] Comparative Example 2 The difference from Example 1 is that Trichoderma harzianum and Trichoderma koningii were not added.

[0047] Comparative Example 3 The difference from Example 1 is that no yam extract was added.

[0048] Comparative Example 4 The difference from Example 1 is that Bacillus subtilis (CGMCC NO.1442) was replaced with Bacillus amyloliquefaciens, which was purchased from Sichuan Lier Crop Science Co., Ltd., and had a viable count greater than 10. 8 cfu / g.

[0049] Example 5 The effects of different compound microbial fertilizers.

[0050] 1. Experimental treatment.

[0051] (1) Test base.

[0052] Located at the Demonier Company base in Daxing District, the trees are of the "Carmen" variety and are 10 years old. The soil organic matter content is 13.2%, total nitrogen is 1.2 g / kg, and the contents of available phosphorus and total potassium are 35.2 mg / kg and 130 mg / kg, respectively.

[0053] (2) Experimental methods: They were randomly divided into the following 9 groups: Treatment 1: Use the compound microbial fertilizer from Example 1; Treatment 2: Use the compound microbial fertilizer from Example 2; Treatment 3: Use the compound microbial fertilizer from Example 3; Treatment 4: Use the compound microbial fertilizer from Example 4; Comparison 1: Using the compound microbial fertilizer from Comparative Example 1; Comparison 2: Using the compound microbial fertilizer from Comparative Example 2; Comparison 3: Using the compound microbial fertilizer from Comparative Example 3; Comparison 4: Using the compound microbial fertilizer from Comparison Example 4; Blank control: Only water was used.

[0054] Each group consisted of 8 rows. The compound microbial fertilizer for each group was mixed with water at a mass ratio of 1:400 to obtain a compound microbial fertilizer solution. After the fruit trees began to sprout flower buds, the solution was applied twice with irrigation water, with an interval of 3 weeks, at a rate of 6 L / acre each time. After the fruit set, the solution was sprayed onto the entire plant every two weeks until the end of the experiment. Other management practices were carried out as usual.

[0055] During the growing season, observe the growth, occurrence of pests and diseases, flowering and fruiting time, and harvest quantity, and keep daily records.

[0056] (3) Postharvest experimental methods.

[0057] After the fruit matured, pears of uniform maturity, free from disease and deformities, were harvested in the morning from both the treatment and control groups. Sixty fruits were collected from each group and transported back to the laboratory immediately within two hours of harvesting. After quality indicators were tested, the fruits were packaged into plastic crates and covered with plastic wrap to prevent dehydration. Storage experiments were conducted at room temperature (24℃±1℃, relative humidity 80%~85%), with daily observation of appearance, rot rate, and soluble solids content.

[0058] 2. Observation content and methods.

[0059] (1) Determination of hardness: GY-4 fruit hardness tester, Zhejiang Top Cloud Agriculture Technology Co., Ltd.; Data recording. Each fruit was rotated twice, and ten fruits were repeated.

[0060] (2) Determination of soluble solids: PAL-1 handheld digital saccharimeter.

[0061] (3) Determination of titratable acid content: potentiometric titration method.

[0062] (4) Rot rate and rot index: The proportion of rotten fruit out of the total fruit count was calculated in 60 fruit groups. Fruits were classified into four levels based on the size of the rotten area: Grade 0: No decay; Grade 1: The rotten area is less than 10% of the fruit's area; Grade 2: Rotten area accounts for 10% to 30% of the fruit area; Level 3: The rotten area is greater than 30% of the fruit area.

[0063] The decay index is calculated using the following formula: Rot index = [Σ(rot level × number of fruits at that level) / (highest rot level × total number of fruits)] × 100.

[0064] 3. Data statistics and graphical analysis.

[0065] All data were statistically analyzed using Excel, the standard error was calculated and plotted; analysis of variance and significance of differences were performed using SPSS software.

[0066] 4. Experimental results and analysis.

[0067] (1) Effect on the hardness of pears.

[0068] Firmness is the most direct indicator of fruit maturity in pear varieties. Before ripening, pears are relatively firm due to the large amount of starch remaining undecomposed. After the respiratory climacteric peak, the firmness gradually decreases, becoming softer and more mushy until it rots and loses its commercial value. At room temperature, the fruit's firmness undergoes a slow softening period from harvest to 3 days, followed by rapid softening from day 3 to day 6. Figure 1The results showed that from day 3 onwards, the firmness of each treatment group decreased more slowly than that of the control groups. In the control groups, the firmness decreased slowly after day 6, at which point the fruit was soft and juicy. Specifically, the firmness of treatment group 1 on day 7 was 3.4 kg·cm². -2 The firmness was equivalent to that of the control group on day 5. This indicates that the treatment group can delay the onset of the respiratory peak, which is beneficial for extending the shelf life of pears.

[0069] (2) Effect on soluble solids of pear.

[0070] As shown in Table 1, the test results on the day of harvest showed that the treated groups significantly increased the soluble solids content, with the best treatment 1 differing from the blank control group by 2.6 Baume degrees. After 6 days of storage at room temperature, the control group reached its peak and then declined, while the treated groups continued to increase without reaching their peak. On the 7th day, treatment 1 was 3.4 Baume degrees higher than the blank control. This indicates that the treatment can significantly improve the quality of pear fruit.

[0071] Table 1. Effects of different treatments on soluble solids in pear.

[0072] (3) Effect on pear decay index.

[0073] The results are shown in Table 2. Under normal temperature storage conditions, starting from day 5, individual cases of rotting appeared in control 4 and the blank control. On day 6, treatments 1 and 2 had not yet rotted, while control 1 and the blank control were the most severely affected. On day 7, treatment 1 still performed the best, with a rot index of only 30.9% of that of the blank control.

[0074] Table 2. Effects of different treatments on the decay index of pears

[0075] It is evident that the compound microbial fertilizer provided by this invention can effectively increase soluble solids, improve fruit firmness, reduce rot rate, and extend freshness and shelf life.

[0076] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A compound microbial fertilizer, characterized in that, The compound microbial fertilizer, by weight, comprises the following raw materials: 0.5-2 parts of growth factor, 1-3 parts of Bacillus subtilis, 1 part of Bacillus cereus, 2-6 parts of Trichoderma harzianum, and 1-3 parts of Trichoderma simonii.

2. The compound microbial fertilizer according to claim 1, characterized in that, The compound microbial fertilizer, by weight, comprises the following raw materials: 1-1.5 parts of growth factor, 2 parts of Bacillus subtilis, 1 part of Bacillus cereus, 3-4 parts of Trichoderma harzianum, and 2 parts of Trichoderma simonii.

3. The compound microbial fertilizer according to claim 1 or 2, characterized in that, The viable counts of Bacillus subtilis, Bacillus cereus, Trichoderma harzianum, and Trichoderma konjac all exceeded 10. 8 cfu / g.

4. The compound microbial fertilizer according to claim 1 or 2, characterized in that, The growth factor is any one or both of yam extract and chitosan.

5. The method of using the compound microbial fertilizer according to any one of claims 1 to 4, characterized in that, Mix the compound microbial fertilizer described in any one of claims 1 to 4 with water to obtain a compound microbial fertilizer solution; after the fruit tree flower buds sprout, apply the compound microbial fertilizer solution twice with water; after fruiting, spray the plants with the compound microbial fertilizer solution.

6. The method of use according to claim 5, characterized in that, The compound microbial fertilizer is mixed with water at a mass ratio of 1:300~500.

7. The method of use according to claim 5, characterized in that, After the fruit tree flower buds sprout, the application rate of compound microbial fertilizer solution is 5-7L / mu.

8. The application of the compound microbial fertilizer according to any one of claims 1 to 4 or the method of use according to any one of claims 5 to 7 in improving the quality of fruits and vegetables.

9. The application of the compound microbial fertilizer according to any one of claims 1 to 4 or the method of use according to any one of claims 5 to 7 in improving the shelf life of fruits and vegetables.

10. The application according to claim 8 or 9, characterized in that, The fruits and vegetables mentioned include pears.

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