Special fertilizer for increasing gum content of Chinese mesona herb

Abstract

The invention belongs to the technical field of Chinese mesona herb, and particularly relates to a special fertilizer for increasing the content of gum in Chinese mesona herb. The special fertilizer for improving the content of the gum in the mesona chinensis comprises a microbial fermentation mixture, and the microbial fermentation mixture is formed by mixing pantoea agglomerans fermentation liquor and bacillus velezensis fermentation liquor. The microbial fermentation mixture in the special fertilizer for improving the gum content of the Chinese mesona herb is formed by mixing the pantoea agglomerans fermentation liquor and the bacillus velezensis fermentation liquor, the gum content of the Chinese mesona herb can be improved by selecting the mixture of the two microbial fermentation liquor, the quality of the Chinese mesona herb can be improved, and the development of the Chinese mesona herb industry is promoted.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of mesona, and particularly relates to a special fertilizer for increasing mesona gum content. BACKGROUND

[0002] Mesona has the effects of clearing heat and quenching thirst and cooling blood, and can relieve heatstroke, heat toxicity, hypertension and joint muscle pain, etc., has high nutritional value and medicinal value, and is a kind of medicinal and edible plant. Mesona polysaccharide, i.e. mesona gum, is the most important component used by mesona, has significant antioxidant function, and is widely applied in the food processing industry, and can be used for making tortoise jelly, mesona jelly, mesona juice and mesona gum, etc. Therefore, it is of great significance to study how to increase the content of mesona gum for the development of mesona industry.

[0003] In the prior art, the quality of mesona can be improved by applying fertilizer. For example, it is disclosed in 'Effect of nitrogen, phosphorus and potassium fertilization on yield and gum content of mesona in pot' that applying reasonable proportions of nitrogen, phosphorus and potassium fertilizer helps to increase the yield and gum content of mesona. However, the large use of nitrogen, phosphorus and potassium fertilizer can easily cause soil acidification and compaction, etc., and has certain disadvantages.

[0004] It is also reported in the prior art that part of microbial agents can prevent and control crop pests, promote crop growth and improve crop quality. However, the effect of using a single microbial agent is limited in actual use. Therefore, it is necessary to screen suitable combinations of microbial formulations.

[0005] The information disclosed in this section of the background art is only intended to increase the understanding of the overall background of the present application, and should not be considered as acknowledging or implying in any form that the information constitutes prior art known to those skilled in the art. SUMMARY

[0006] The purpose of the present application is to provide a special fertilizer for increasing mesona gum content, so as to solve the problem of limited effect of using a single microbial agent in the background art.

[0007] To achieve the above-mentioned purpose, the present application provides the following technical solutions. The special fertilizer for increasing mesona gum content comprises a microbial fermentation mixture, and the microbial fermentation mixture is mixed by a pantoea agglomerans fermentation liquid and a bacillus velezensis fermentation liquid.

[0008] Further, the pantoea agglomerans is purchased from Guangdong Microbial Culture Collection Center, and the preservation number is GDMCC NO.812725.

[0009] Further, the bacillus velezensis is purchased from China General Microbiological Culture Collection Center, and the preservation number is CGMCC NO.1.8805.

[0010] Furthermore, the mass ratio of the Pantotheca agglomerata fermentation broth to the Bacillus vesiculosus fermentation broth is 1-3:5-1.

[0011] Furthermore, the preparation method of the Pantotheca agglomerata fermentation broth is as follows: a single colony of Pantotheca agglomerata is inoculated into beef extract peptone liquid medium and cultured at 28°C and 180 r / min with shaking until OD reaches 180°C. 600 The value was 0.8, so fresh beef extract peptone liquid medium was added to adjust the bacterial count to 3.2 × 10⁸. 7 The cfu / mL concentration was used to obtain the clump-forming pantothenic acid fermentation broth.

[0012] Furthermore, the preparation method of the Bacillus vesicularis fermentation broth is as follows: Bacillus vesicularis is inoculated into beef extract peptone liquid culture medium and cultured at 30°C with shaking at 180 r / min until OD reaches 180°C. 600 The value was 0.8, so fresh beef extract peptone liquid medium was added to adjust the bacterial count to 1.0 × 10⁸. 8 The fermentation broth of Bacillus belye was obtained by dissolving cfu / mL in the solution.

[0013] Furthermore, the formulation of the beef extract peptone liquid culture medium is as follows: 5 g / L peptone, 3 g / L beef extract, 5 g / L sodium chloride, and pH 7.2.

[0014] This invention also provides the application of the special fertilizer for increasing the gum content of grass jelly in increasing the gum content of grass jelly.

[0015] This invention also provides the application of the special fertilizer for increasing the gum content of jelly grass in antagonizing jelly grass root-knot nematodes.

[0016] Compared with the prior art, the present invention has the following beneficial effects: (1) The microbial fermentation mixture in the special fertilizer for improving the gum content of grass jelly of the present invention is composed of a mixture of Pantotheca fermentation liquid and Bacillus vesiculosus fermentation liquid. Using this mixture of two microbial fermentation liquids can increase the gum content of grass jelly, which helps to improve the quality of grass jelly and promote the development of the grass jelly industry.

[0017] (2) The microbial fermentation mixture of the present invention, which is composed of Pantotheca fermentation liquid and Bacillus vesiculosus fermentation liquid, can improve the antagonistic effect on Mesona chinensis root-knot nematode, effectively resist the damage of Mesona chinensis by root-knot nematode, and can provide support for the development of microbial fertilizers or biocontrol agents for the prevention and control of Mesona chinensis root-knot nematode. Detailed Implementation

[0018] The technical solution of this invention patent will be clearly and completely described below. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without inventive effort are within the scope of protection of this invention.

[0019] 1. Materials and Methods 1.1 Test culture medium Beef extract peptone liquid culture medium: peptone 5 g / L, beef extract 3 g / L, sodium chloride 5 g / L, pH 7.2; 1.2 Fermentation broth of the tested microorganism Pantothecin fermentation broth: Pantothecin bacteria purchased from Guangdong Provincial Microbial Culture Collection Center Pantoea vagans (GDMCC No. 812725) A single colony was inoculated into beef extract peptone liquid medium and cultured at 28°C with shaking at 180 rpm until OD reached. 600 The value was 0.8, and then fresh beef extract peptone liquid medium was added to adjust the bacterial count to 3.2 × 10⁻⁸. 7 The cfu / mL concentration was used to obtain the clump-forming pantothenic acid fermentation broth.

[0020] Bacillus velezensis fermentation broth: Single colonies of Bacillus velezensis (accession number GDMCC NO. 1.8805), purchased from the China General Microbiological Culture Collection Center, were inoculated into beef extract peptone liquid medium and cultured at 30°C with shaking at 180 rpm until OD reached the target concentration. 600 The value was 0.8, and then fresh beef extract peptone liquid medium was added to adjust the bacterial count to 1.0 × 10⁸. 8 The fermentation broth of Bacillus belye was obtained by dissolving cfu / mL in the solution.

[0021] 1.3 Antagonistic effect of microbial fermentation broth on root-knot nematodes of Mesona chinensis Referring to "NT / Y 1833.1-2009 Guidelines for Indoor Bioassay Testing of Pesticides - Nematicides - Part 1: Inhibition of Plant Pathogenic Nematodes - Immersion Method", the specific procedure is as follows: Select southern root-knot nematodes from the roots of *Mesona chinensis* (…). Meloidogyne incognita The eggs were hatched and cultured in the laboratory. Second-instar larvae of the same instar were resuspended in water to a concentration of 150 larvae / mL for later use.

[0022] 1.3.1 Antagonistic effect of single agent of microbial fermentation broth on root-knot nematodes of Mesona chinensis The fermentation broth of the tested microorganisms was diluted with sterile water to create five mass concentration gradients. An equal volume of the diluted fermentation broth was mixed thoroughly with a root-knot nematode suspension. 3 mL of this mixture was pipetted into six wells of a 24-well biochemical assay plate, capped, and incubated at 25°C. A sterile water treatment was used as a blank control. Each treatment was repeated four times, with each repetition involving six wells of the 24-well biochemical assay plate.

[0023] After 24 hours of culture, 1 mL of the mixture from each treatment was taken and the mortality of nematodes was observed under a dissecting microscope. Each replicate included 120 nematodes, and the number of main nematodes and the number of dead nematodes were recorded. Based on the data, the mortality rate and corrected mortality rate for each treatment were calculated. The nematode mortality criterion was: the nematodes were stiff and could not bend or move when touched with a hairpin or bamboo needle.

[0024]

[0025] In the above formula: P -- Mortality rate, in percentages. K --The number of dead insects; N --Total number of insects treated.

[0026]

[0027] In the above formula: P 1 -- Corrected mortality rate, in % P t --Case-level mortality rate, in % P 0 -- Mortality rate in the blank control group, in percent.

[0028] Data statistical analysis was performed using DPS software to calculate the virulence regression equations and EC values ​​for different treatments. 50 Values ​​(mg / L), results are shown in Table 1.

[0029] Table 1. Antagonistic effects of single-agent microbial fermentation broth against root-knot nematodes of Mesona chinensis

[0030] Table 1 shows that the fermentation broths of Pantotheca acuminata and Bacillus vesiculosus have different effects on the EC50 of Mesona chinensis root-knot nematodes. 50 The values ​​were 143.263 mg / L and 85.910 mg / L, respectively.

[0031] 1.3.2 Antagonistic effect of microbial fermentation broth mixture on root-knot nematodes of Mesona chinensis The fermentation broth of the tested microorganisms was prepared into multiple formulations, and then diluted with sterile water to create five mass concentration gradients. Following the method described in "1.3.1", the toxicity regression equation for each formulation and the EC50 against *Gnaphalium affine* root-knot nematode were obtained. 50The values ​​were then calculated using the Sun Yunpei method to determine the co-toxicity coefficient (CTC). Based on the CTC, the type of effect of the mixed solution on the root-knot nematode of the grass was evaluated: CTC≤80 indicates antagonistic effect, 80<CTC<120 indicates additive effect, and CTC≥120 indicates synergistic effect. The results are shown in Table 2.

[0032] Table 2. Antagonistic effect of microbial fermentation broth mixture on root-knot nematodes of Mesona chinensis

[0033] Table 2 shows that when the fermentation broth of *Pantotheca acuminata* and *Bacillus vesicularis* is mixed at a mass ratio of 1:24-1, the combined effects against *Mesona chinensis* root-knot nematodes are not entirely the same. Within a mass ratio range of 1:20-8, the co-toxicity coefficient of the two antagonistic agents against *Mesona chinensis* root-knot nematodes is greater than 120, indicating a synergistic effect; while at mass ratios of 1:24, 1:4, and 1:1, an additive effect is observed. This indicates that the microbial fermentation mixture of the present invention can improve the antagonistic effect against *Mesona chinensis* root-knot nematodes, effectively resisting the damage caused by root-knot nematodes to *Mesona chinensis*, and can provide support for the development of microbial fertilizers or biocontrol agents for the control of *Mesona chinensis* root-knot nematodes.

[0034] 2. Effect of microbial fermentation broth on the gum content of jelly grass 2.1 Fermentation broth of the test microorganism Same as "1.2 Fermentation broth of the test microorganism".

[0035] 2.2 Test Methods The experiment was conducted in multiple small cells, each with an area of ​​50m². 2 The randomized controlled trials were divided into four groups: study group 1, study group 2, study group 3, and a blank control group. The plots were tilled and raised into beds, 120cm wide between beds, with each bed 100cm wide and 30cm high. A 15-15-15 NPK compound fertilizer was spread on the beds, mixed with fine soil, and seedlings of *Mesona chinensis* (approximately 25cm tall) were transplanted onto the beds at a spacing of 30cm x 40cm. The soil around the roots was pressed down, and the plants were thoroughly watered. Treatments were administered at 7, 14, and 30 days after transplanting: Research Group 1: Root irrigation with Panax notoginseng fermentation broth, 25 mL / plant; Research Group 2: Fermentation broth of Bacillus berberis, 25 mL / strain; Study Group 3: Root irrigation consisted of a microbial fermentation mixture of Pantotheca acuminata fermentation broth and Bacillus vesiculosus fermentation broth at a mass ratio of 1:16, 25 mL / plant; Blank control group: roots were irrigated with clean water, 25 mL / plant.

[0036] The remaining water and fertilizer management was the same for all plots. Harvesting was done before the flower buds appeared. Twenty plants were randomly selected from each plot, dried to constant weight, and their polysaccharide content was determined. The results are shown in Table 3.

[0037] Table 3. Effects of microbial fermentation broth on the content of grass jelly gum.

[0038] Table 3 shows that, compared with the blank control, applying microbial fermentation broth during the cultivation of *Mesona chinensis* can increase the polysaccharide content (i.e., *Mesona chinensis* gum content), with study group 3 showing the highest polysaccharide content. This indicates that the microbial fermentation mixture of the present invention can increase the *Mesona chinensis* gum content.

[0039] In summary, the microbial fermentation mixture of the present invention can enhance the antagonistic effect against root-knot nematodes in Mesona chinensis, effectively resisting the damage of root-knot nematodes to Mesona chinensis, and can provide support for the development of microbial fertilizers or biocontrol agents for the prevention and control of root-knot nematodes in Mesona chinensis; in addition, the microbial fermentation mixture of the present invention can increase the gum content of Mesona chinensis and improve the quality of Mesona chinensis.

[0040] 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 special fertilizer for increasing the gum content of grass jelly, characterized in that, The special fertilizer for increasing the gum content of grass jelly includes a microbial fermentation mixture, which is composed of a mixture of Pantotheca acuminata fermentation broth and Bacillus vesiculosus fermentation broth.

2. The special fertilizer for increasing the gum content of jelly grass according to claim 1, characterized in that, The aforementioned Pantotheca acuminata was purchased from the Guangdong Provincial Center for Microbial Culture Collection, with accession number GDMCC NO. 812725.

3. The special fertilizer for increasing the gum content of jelly grass according to claim 1, characterized in that, The Bacillus belyssus was purchased from the China General Microbiological Culture Collection Center, accession number CGMCC NO. 1.8805.

4. The special fertilizer for increasing the gum content of jelly grass according to claim 1, characterized in that, The mass ratio of the Pantotheca agglomerata fermentation broth to the Bacillus vesiculosus fermentation broth is 1-3:5-1.

5. The special fertilizer for increasing the gum content of jelly grass according to claim 1, characterized in that, The preparation method of the Pantotheca agglomerata fermentation broth is as follows: a single colony of Pantotheca agglomerata is inoculated into beef extract peptone liquid medium and cultured at 28°C and 180 r / min with shaking until OD reaches 180°C. 600 The value was 0.8, so fresh beef extract peptone liquid medium was added to adjust the bacterial count to 3.2 × 10⁸. 7 The cfu / mL concentration was used to obtain the clump-forming pantothenic acid fermentation broth.

6. The special fertilizer for increasing the gum content of jelly grass according to claim 1, characterized in that, The preparation method of the Bacillus belysin fermentation broth is as follows: Bacillus belysin is inoculated into beef extract peptone liquid culture medium and cultured at 30°C and 180 r / min with shaking until OD. 600 The value was 0.8, so fresh beef extract peptone liquid medium was added to adjust the bacterial count to 1.0 × 10⁸. 8 The fermentation broth of Bacillus belye was obtained by dissolving cfu / mL in the solution.

7. The special fertilizer for increasing the gum content of jelly grass according to claim 5 or claim 6, characterized in that, The formula for the beef extract peptone liquid culture medium is as follows: 5 g / L peptone, 3 g / L beef extract, 5 g / L sodium chloride, pH 7.

2.

8. The application of the special fertilizer for increasing the gum content of grass jelly as described in claim 1 in increasing the gum content of grass jelly.

9. The application of the special fertilizer for increasing the gum content of jelly grass as described in claim 1 in antagonizing jelly grass root-knot nematodes.

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