Method for improving quality of mesona acuminata and application thereof

Abstract

This invention belongs to the field of Mesona chinensis cultivation technology, specifically relating to a method for improving the quality of Mesona chinensis and its application. A method for improving the quality of Mesona chinensis includes the following steps: S1. Site selection and land preparation; S2. Bed preparation and fertilization; S3. Mulching and transplanting; S4. Water and fertilizer management: irrigate the roots with a compound bacterial solution once each at 10, 20, and 40 days after transplanting, using 15-20 mL per seedling; the compound bacterial solution is a mixture of Beauveria bassiana bacterial solution, Phytotrichum lacerum GXMS1 bacterial solution, and Trichoderma harzianum bacterial solution in a volume ratio of 1-3:1:1-8; S5. Pest and disease control; S6. Timely harvesting. This invention involves applying a compound bacterial solution, composed of Beauveria bassiana, Phytotrichum laceratum GXMS1, and Trichoderma harzianum in a specific volume ratio, during the cultivation of Mesona chinensis. This significantly increases the content of polysaccharides, total flavonoids, and polyphenols in Mesona chinensis plants, which helps improve the quality of Mesona chinensis and can support the green, environmentally friendly, and high-quality production of the Mesona chinensis industry.

Description

Technical Field

[0001] This invention belongs to the field of Mesona chinensis cultivation technology, specifically relating to a method for improving the quality of Mesona chinensis and its application. Background Technology

[0002] Mesona chinensis, also known as fairy grass, is an annual herb belonging to the genus Mesona in the Lamiaceae family. It has high nutritional and medicinal value. Mesona chinensis is rich in active ingredients such as polysaccharides, flavonoids, and phenolic acids. In the food industry, it is widely used in the processing of traditional foods such as herbal tea, jelly, and turtle jelly. In the medical field, it has various pharmacological activities such as antioxidation, hypoglycemia, and anti-inflammation.

[0003] With increasing public attention to green and healthy foods, the market demand for high-quality grass jelly raw materials is continuously growing. However, traditional grass jelly cultivation relies heavily on the application of nitrogen, phosphorus, and potassium compound fertilizers. While this can temporarily increase biomass yield, excessive or excessive use of chemical fertilizers can negatively impact the quality of the grass jelly, causing soil compaction and reducing soil fertility, thus affecting the quality of the grass jelly. Current technologies, such as foliar spraying with amino acid chelated fertilizers or trace elements, can temporarily increase the content of polysaccharides and other internal components in the plant, but chemical residues can exacerbate environmental pollution and directly affect the food safety of grass jelly products.

[0004] Existing technologies also disclose methods to improve the quality of grass jelly through intercropping. For example, CN117859607A discloses a cultivation method for intercropping grass jelly with sugarcane in dryland areas. The total flavonoid, total sugar, and polysaccharide content of intercropped grass jelly is significantly higher than that of monoculture. However, this intercropping model is complex to operate and requires the control of multiple planting operations.

[0005] Furthermore, studies have shown that microbial inoculants can increase the content of bioactive substances in plants by regulating plant metabolic pathways. Microbial inoculants are also environmentally friendly and green. However, the function of a single microbial strain is limited. Reasonable mixing of microbial strains can further increase the content of bioactive substances in plants, but not every mixture of two or more microbial strains will achieve a synergistic effect.

[0006] There are currently no reports on using microbial agents to improve the quality of grass jelly. Summary of the Invention

[0007] The purpose of this invention is to provide a method for improving the quality of grass jelly and its application.

[0008] To achieve the above objectives, the present invention provides the following technical solution:

[0009] A method for improving the quality of grass jelly includes the following steps:

[0010] S1. Site selection and land preparation

[0011] Choose sandy loam soil, plow to a depth of 25-35cm, break up the soil, and let it dry for 30-40 days;

[0012] S2. Fertilizing and ridging

[0013] The width between ridges is 120-135cm, the ridge surface is 100-110cm wide, and the ridge height is 30-35cm; spread well-rotted farmyard manure on the ridge surface and mix it with fine soil;

[0014] S3. Mulching and Planting

[0015] Cover the raised bed with plastic film and punch holes in the film according to the planned planting spacing; plant the 20-25cm tall grass seedlings in the holes, press the soil around the roots firmly, and water thoroughly.

[0016] S4. Water and Fertilizer Management

[0017] The root of each seedling was irrigated with a compound bacterial solution at 10, 20 and 40 days after transplanting. The compound bacterial solution was prepared by mixing Beauveria bassiana, Phytotrichum lacerum GXMS1 and Trichoderma harzianum in a volume ratio of 1-3:1:1-8.

[0018] S5. Pest and Disease Control

[0019] Prevention is the primary focus, with comprehensive prevention and control measures.

[0020] S6. Timely harvesting

[0021] Harvest before the flower buds appear.

[0022] More specifically, in step S2, the amount of well-rotted farmyard manure used is 1500-2000 kg / mu.

[0023] More specifically, in step S3, the planting spacing is 30cm × 40cm; 2 plants are planted in each hole.

[0024] More specifically, in step S4, the bacterial content of the Beauveria bassiana suspension, the Phytotrichum lacerum GXMS1 suspension, and the Trichoderma harzianum suspension is all 5.0 × 10⁻⁶. 5 cfu / mL.

[0025] More specifically, the preparation method of the Beauveria bassiana suspension is as follows: Beauveria bassiana is inoculated onto a PDA medium plate and cultured at 25°C for 10 days; 10 mL of sterile water is added to the plate, and the spores and sterile water are mixed evenly with a sterile glass rod; the medium and mycelium are filtered out through four layers of sterile gauze to obtain a fresh spore suspension; the spore concentration in the spore suspension is determined using a hemocytometer, and the spore concentration is adjusted to 5.0 × 10⁻⁶ with sterile water. 5 cfu / mL.

[0026] More specifically, the preparation method of the *Phenothizoctonia solani* GXMS1 bacterial suspension is as follows: *Phenothizoctonia solani* GXMS1 is inoculated onto a PDA medium plate and cultured at 28°C for 13 days; 10 mL of sterile water is added to the plate, and the spores and sterile water are mixed evenly with a sterile glass rod; the medium and mycelium are filtered out through four layers of sterile gauze to obtain a fresh spore suspension; the spore concentration in the spore suspension is determined using a hemocytometer, and the spore concentration is adjusted to 5.0 × 10⁻⁶ with sterile water. 5 cfu / mL.

[0027] More specifically, the preparation method of the *Trichoderma harzianum* suspension is as follows: *Trichoderma harzianum* is inoculated onto a PDA medium plate and cultured at 25°C for 7 days; 10 mL of sterile water containing 0.05% Tween-80 is added to the plate, and the spores and sterile water are mixed evenly with a sterile glass rod; the medium and mycelium are filtered out through four layers of sterile gauze to obtain a fresh spore suspension; the spore concentration in the spore suspension is determined using a hemocytometer, and the spore concentration is adjusted to 5.0 × 10⁻⁶ with sterile water. 5 cfu / mL.

[0028] The present invention also provides the method for improving the quality of grass jelly or the application of the compound bacterial solution in improving the quality of grass jelly.

[0029] The present invention also provides the method for improving the quality of Mesona chinensis or the application of the compound bacterial solution in improving the content of polysaccharides, total phenols and total flavonoids in Mesona chinensis.

[0030] Compared with the prior art, the present invention has the following beneficial effects:

[0031] This invention involves applying a compound bacterial solution, composed of Beauveria bassiana, Phytotrichum laceratum GXMS1, and Trichoderma harzianum in a specific volume ratio, during the cultivation of Mesona chinensis. This significantly increases the content of polysaccharides, total flavonoids, and total phenols in Mesona chinensis plants, which helps improve the quality of Mesona chinensis and can support the green, environmentally friendly, and high-quality production of the Mesona chinensis industry. Detailed Implementation

[0032] The technical solution of this invention patent will be clearly and completely described below. Obviously, the described embodiments are some embodiments of this invention, but not all embodiments.

[0033] In this application, prior to the experiment, it was verified through testing that there were no antagonistic effects between any of the three strains: *Beauveria bassiana*, *Founculia laceratum* GXMS1, and *Trichoderma harzianum*. Furthermore, in this application, the *Beauveria bassiana* stock solution was 50 billion spores / gram (purchased from Hebei Zhongbaolv Agricultural Technology Co., Ltd.); the *Founculia laceratum* GXMS1 stock solution was 15 million CFU / gram (Sichuan Jinzhu Ecological Agriculture Technology Co., Ltd.); and the *Trichoderma harzianum* stock solution was 30 billion CFU / gram (Byver Corporation, USA).

[0034] 1. Effects of different bacterial solutions on the quality of grass jelly

[0035] 1.1 Preparation of Beauveria bassiana inoculum

[0036] Beauveria bassiana was inoculated onto PDA agar plates and incubated at 25°C for 10 days. 10 mL of sterile water was added to the plates, and the spores and water were mixed thoroughly with a sterile glass rod. The culture medium and mycelium were filtered out using four layers of sterile gauze to obtain a fresh spore suspension. The spore concentration in the suspension was determined using a hemocytometer, and the concentration was adjusted to 5.0 × 10⁻⁶ spores with sterile water. 5 cfu / mL.

[0037] 1.2 Preparation of *Porphyromonas laceratus* GXMS1 bacterial culture

[0038] *Phellinus cereus* GXMS1 was inoculated onto PDA agar plates and incubated at 28°C for 13 days. 10 mL of sterile water was added to the plates, and the spores and water were mixed thoroughly with a sterile glass rod. The culture medium and mycelium were filtered out using four layers of sterile gauze to obtain a fresh spore suspension. The spore concentration in the suspension was determined using a hemocytometer, and the concentration was adjusted to 5.0 × 10⁻⁶ spores with sterile water. 5 cfu / mL.

[0039] 1.3 Preparation of Trichoderma harzianum inoculum

[0040] Trichoderma harzianum was inoculated onto PDA agar plates and incubated at 25°C for 7 days. 10 mL of sterile water containing 0.05% Tween-80 was added to the plates, and the spores and sterile water were mixed thoroughly with a sterile glass rod. The culture medium and mycelium were filtered out using four layers of sterile gauze to obtain a fresh spore suspension. The spore concentration in the suspension was determined using a hemocytometer, and the spore concentration was adjusted to 5.0 × 10⁻⁶ with sterile water. 5 cfu / mL.

[0041] 1.4 Test Methods

[0042] 1.4.1 Cultivation methods of Mesona chinensis

[0043] S1. Site selection and land preparation

[0044] Select sandy loam soil plots at the experimental base of the Guangxi South Subtropical Agricultural Science Research Institute in Longzhou County, Chongzuo City, Guangxi, plow to a depth of 25cm, break up the soil, and let it dry for 35 days.

[0045] S2. Fertilizing and ridging

[0046] The width between ridges is 130cm, the ridge surface is 100cm wide, and the ridge height is 30cm. Spread the well-rotted farmyard manure on the ridge surface and mix it with fine soil. The well-rotted farmyard manure is chicken manure organic fertilizer purchased from Hebei Sanpin Fertilizer Co., Ltd. (implementation standard: NY525-2012, nitrogen, phosphorus and potassium ≥5%, organic matter ≥45%), and the dosage is 1800kg / mu.

[0047] S3. Mulching and Planting

[0048] Cover the raised bed with plastic film and make holes in the film according to the planned planting spacing (30cm×40cm). In mid-March, plant the 25cm tall seedlings of "Lingshan Big Leaf" grass in the holes, with 2 seedlings per hole, press the soil around the roots firmly, and water thoroughly.

[0049] S4. Water and Fertilizer Management

[0050] The roots of each seedling were irrigated with bacterial solution once on the 10th, 20th and 40th day after transplanting of the grass jelly, with 20 mL of bacterial solution per seedling; the bacterial solution is shown in Table 1.

[0051] Table 1 Different bacterial solutions

[0052]

[0053] S5. Pest and Disease Control

[0054] Prevention is the primary focus, with comprehensive prevention and control measures.

[0055] S6. Timely harvesting

[0056] Harvest before the flower buds appear.

[0057] 1.4.2 Specific Experimental Setup

[0058] The experiment was conducted in multiple small cells, each with an area of ​​50m². 2 The cells were randomly assigned to nine groups: Studies 1-9. All plots were identical except for step S4 (water and fertilizer management). Specifically, Studies 1-8 applied bacterial solution 1-8 for root irrigation, while Study 9 received an equal volume of water as a control. Each group had three replicates.

[0059] 1.5 Quality Characteristics

[0060] When harvesting *Mesona chinensis*, 25 plants were randomly selected from each plot, dried at 85℃ to constant weight, and then pulverized through an 80-mesh sieve. The contents of polysaccharides, total flavonoids, and total phenols were determined according to the study on "Selection, Planting, and Fermentation of High-Colloidal *Mesona chinensis*" (Yan Yueyue et al., 2023), and the results are shown in Table 2.

[0061] Table 2. Effects of different bacterial solutions on the quality of *Gynostemma pentaphyllum*

[0062] Research Group 1 70.1 52.1 22.8 Research Group 2 65.6 58.3 23.6 Research Group 3 76.9 61.2 24.1 Research Group 4 79.4 64.8 28.1 Research Group 5 83.0 66.6 30.5 Research Group 6 84.5 72.3 32.4 Research Group 7 98.4 75.8 36.3 Research Group 8 100.6 79.4 39.5 Research Group 9 62.3 46.4 19.2

[0063] As shown in Tables 1 and 2, compared with the blank control group 9, the application of Beauveria bassiana, Phytotrichum lacerum GXMS1, and Trichoderma harzianum inoculum solutions all increased the content of polysaccharides, total flavonoids, and total phenols in Mesona chinensis plants to a certain extent. This indicates that Beauveria bassiana, Phytotrichum lacerum GXMS1, and Trichoderma harzianum inoculum solutions help improve the quality of Mesona chinensis.

[0064] Compared with single-strain solutions, mixing Beauveria bassiana, Phytotrichum lacerum GXMS1, and Trichoderma harzianum in pairs can further increase the content of polysaccharides, total flavonoids, and total phenols in Mesona chinensis plants, but the improvement effect is not significant.

[0065] Compared with the two mixed bacterial solutions, the content of polysaccharides, total flavonoids and polyphenols in the grass plant was significantly increased after mixing the three bacterial solutions: Beauveria bassiana, Phytotrichum lacerum GXMS1, and Trichoderma harzianum. The improvement effect was particularly significant when the mass ratio was 2:1:6.

[0066] In summary, the application of a compound bacterial solution composed of Beauveria bassiana, Phytotrichum lacerum GXMS1, and Trichoderma harzianum during the cultivation of Mesona chinensis can significantly increase the content of polysaccharides, total flavonoids, and total phenols in Mesona chinensis plants, which helps to improve the quality of Mesona chinensis.

[0067] The foregoing description of specific exemplary embodiments of the invention is for illustrative and explanatory purposes. These descriptions are not intended to limit the invention to the precise forms disclosed, and it will be apparent that many changes and variations can be made in accordance with the foregoing teachings. The exemplary embodiments were chosen and described in order to explain the specific principles of the invention and its practical application, thereby enabling those skilled in the art to implement and utilize various different exemplary embodiments of the invention, as well as various different choices and variations. The scope of the invention is intended to be defined by the claims and their equivalents.

Claims

1. A method for improving the quality of *Mesona chinensis* (a type of herb) and its application in increasing the content of polysaccharides, total phenols, and total flavonoids in *Mesona chinensis*, characterized in that... The method for improving the quality of grass jelly includes the following steps: S1. Site selection and land preparation Choose sandy loam soil, plow to a depth of 25-35cm, break up the soil, and let it dry for 30-40 days; S2. Raising beds and applying fertilizer The width between ridges is 120-135cm, the ridge surface is 100-110cm wide, and the ridge height is 30-35cm; spread well-rotted farmyard manure on the ridge surface and mix it with fine soil; S3. Mulching and Planting Cover the raised bed with plastic film and punch holes in the film according to the planned planting spacing; plant the 20-25cm tall grass seedlings in the holes, press the soil around the roots firmly, and water thoroughly. S4. Water and Fertilizer Management The roots of each seedling were drenched with a compound bacterial solution at 10, 20, and 40 days after transplanting. The solution consisted of 15-20 mL of the solution mixed with *Beauveria bassiana*, *Inonotus cereus* GXMS1, and *Trichoderma harzianum* in a volume ratio of 1-3:1:1-8. The bacterial content of each of these solutions was 5.0 × 10⁻⁶. 5 cfu / mL; S5. Pest and Disease Control Prevention is the primary focus, with comprehensive prevention and control measures. S6. Harvest in a timely manner Harvest before the flower buds appear.

2. The application according to claim 1, characterized in that, In step S2, the amount of well-rotted farmyard manure used is 1500-2000 kg / mu.

3. The application according to claim 1, characterized in that, In step S3, the planting spacing is 30cm × 40cm; 2 plants are planted in each hole.

4. The application according to claim 1, characterized in that, The preparation method of the Beauveria bassiana suspension is as follows: Beauveria bassiana is inoculated onto a PDA medium plate and cultured at 25℃ for 10 days; 10 mL of sterile water is added to the plate, and the spores and sterile water are mixed evenly with a sterile glass rod; the medium and mycelium are filtered out through four layers of sterile gauze to obtain a fresh spore suspension; the spore concentration in the spore suspension is determined using a hemocytometer, and the spore concentration is adjusted to 5.0 × 10⁻⁶ with sterile water. 5 cfu / mL.

5. The application according to claim 1, characterized in that, The preparation method of the *Phenothizoctonia solani* GXMS1 bacterial suspension is as follows: *Phenothizoctonia solani* GXMS1 is inoculated onto a PDA medium plate and cultured at 28℃ for 13 days; 10 mL of sterile water is added to the plate, and the spores and sterile water are mixed evenly with a sterile glass rod; the medium and mycelium are filtered out through four layers of sterile gauze to obtain a fresh spore suspension; the spore concentration in the spore suspension is determined using a hemocytometer, and the spore concentration is adjusted to 5.0 × 10⁻⁶ with sterile water. 5 cfu / mL.

6. The application according to claim 1, characterized in that, The preparation method of the *Trichoderma harzianum* suspension is as follows: *Trichoderma harzianum* is inoculated onto a PDA medium plate and cultured at 25℃ for 7 days; 10 mL of sterile water containing 0.05% Tween-80 is added to the plate, and the spores and sterile water are mixed evenly with a sterile glass rod. The medium and mycelium are filtered out through four layers of sterile gauze to obtain a fresh spore suspension; the spore concentration in the spore suspension is determined using a hemocytometer, and the spore concentration is adjusted to 5.0 × 10⁻⁶ with sterile water. 5 cfu / mL.

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