Salvia miltiorrhiza quality improving and yield increasing method based on straw source smoke water and bacillus amyloliquefaciens

Abstract

The invention discloses a quality-improving and yield-increasing method for salvia miltiorrhiza based on straw source smoke water and bacillus amyloliquefaciens, and relates to the technical field of planting methods. According to the method, the tobacco water and the bacillus amyloliquefaciens are compounded to obtain a mixed solution, and the mixed solution is adopted to perform root soaking and foliage spraying on the salvia miltiorrhiza, so that the yield and the quality of the salvia miltiorrhiza are improved, and particularly, the tobacco water and the bacillus amyloliquefaciens are combined for use; the single plant accumulation amounts of the tanshinone I, the tanshinone IIA, the cryptotanshinone and the salvianolic acid B are increased by 20.33%, 12.92%, 26.53% and 102.01% compared with those of a group without treatment, and the bacillus amyloliquefaciens has a synergistic effect on increasing the content of medicinal components in the radix salviae miltiorrhizae. In addition, by means of the method, the yield and agronomic characters of the salvia miltiorrhiza can be improved, specifically, the weight, SPAD value, net photosynthetic rate, stomatal conductance and nitrogen phosphorus and potassium accumulation amount of the salvia miltiorrhiza roots are improved.

Description

Technical Field

[0001] The invention relates to the field of planting methods and application technologies, and in particular to a method for improving the quality and increasing the yield of salvia miltiorrhiza based on straw-derived smoke water and Bacillus amyloliquefaciens. Background Art

[0002] Salvia miltiorrhiza Bunge belongs to the genus Salvia of the family Labiatae. It is a perennial herb mainly used as medicine with roots and rhizomes. It is a drug included in the Pharmacopoeia of the People's Republic of China (2020 edition). Because the active ingredients in the rhizomes have extremely high medicinal value, they are widely used clinically to treat coronary heart disease, angina pectoris and other diseases. However, wild Salvia miltiorrhiza grows slowly and has low yields. In recent years, due to excessive exploitation of huge market demand and habitat loss caused by human activities, the wild population resources of Salvia miltiorrhiza have decreased sharply, and the yield and quality of Salvia miltiorrhiza have been affected, and can no longer meet market demand. Therefore, many areas have begun to cultivate it artificially, but the yield and quality of artificially cultivated Salvia miltiorrhiza need to be improved. The development and quality issues of the Salvia miltiorrhiza industry have attracted widespread attention at home and abroad.

[0003] Plant-derived smoke water is made by dissolving smoke produced by temperature-controlled pyrolysis of plant materials in water. It can promote seed germination and plant growth, improve crop resistance to adversity, and has the advantages of high efficiency, safety, low cost and environmental friendliness. In recent years, smoke water has been applied to a variety of crop research and has shown significant results. Bacillus amyloliquefaciens is named for its ability to produce amylase to decompose starch. During its growth, it can produce a series of metabolites that inhibit pathogens. It has the effect of promoting growth and resisting adversity in crop production, and has the advantages of being green and environmentally friendly. It has broad application prospects in agriculture.

[0004] In the prior art, the paper "Study on the Accumulation and Mechanism of Salvianolic Acids in Salvia Miltiorrhiza Hairy Roots Regulated by Plant-derived Smoke Water" (Sun Hui, Jinan University, 2021) discloses the use of plant-derived smoke water to suspend Salvia Miltiorrhiza hairy roots, and the accumulation of salvianolic acid compounds can be promoted by treating them with the active substance KAR1 in the plant-derived smoke water. The paper "Effects of Straw-derived Smoke Water and Bacillus Amyloliquefaciens Treatment on Agronomy, Yield and Quality Shape of Corn" (Xu Jiacheng, Shandong Agricultural University, 2023) discloses the use of smoke water and Bacillus amyloliquefaciens in combination to prime corn seeds to improve corn seed vitality, photosynthetic parameters, root system and quality traits, but corn and Salvia Miltiorrhiza are two plants from different families, and the synthesis mechanism of their active ingredients is completely different from that of the active ingredients in Salvia Miltiorrhiza.

[0005] Therefore, it is necessary to use tobacco water and Bacillus amyloliquefaciens to increase the yield of Salvia miltiorrhiza and the content of its active ingredients. Summary of the invention

[0006] In view of the above-mentioned prior art, the purpose of the present invention is to provide a method for improving the quality and yield of Salvia miltiorrhiza based on straw-source tobacco water and Bacillus amyloliquefaciens. The present invention uses tobacco water and Bacillus amyloliquefaciens to treat Salvia miltiorrhiza by root soaking and foliar spraying, which is beneficial to improve the yield and quality of Salvia miltiorrhiza. Specifically, by combining tobacco water and Bacillus amyloliquefaciens, the single-plant accumulation of tanshinone I, tanshinone IIA, cryptotanshinone and salvianolic acid B is increased by 20.33%, 12.92%, 26.53% and 102.01% compared with the untreated group. Tobacco water and Bacillus amyloliquefaciens have a synergistic effect in increasing the content of the active ingredients in Salvia miltiorrhiza. In addition, the method of the present invention can also improve the yield and agronomic traits of Salvia miltiorrhiza, which is specifically manifested in improving the root weight, SPAD value, net photosynthetic rate, stomatal conductance and nitrogen, phosphorus and potassium accumulation of Salvia miltiorrhiza.

[0007] To achieve the above object, the present invention adopts the following technical solution:

[0008] The present invention provides a method for improving the quality and increasing the yield of Salvia miltiorrhiza based on straw-derived tobacco water and Bacillus amyloliquefaciens, comprising the following steps:

[0009] (1) mixing the diluted tobacco water solution and the diluted Bacillus amyloliquefaciens solution to obtain a mixed solution, and using the mixed solution to perform root immersion treatment on Salvia miltiorrhiza seedlings;

[0010] (2) After the root-immersion treatment of the Salvia miltiorrhiza seedlings is transplanted to the field, the mixed solution is sprayed on the leaves of the Salvia miltiorrhiza during its growth and development period.

[0011] Preferably, the tobacco water dilution liquid is obtained by diluting the tobacco water stock liquid by 500 times.

[0012] Further preferably, the tobacco liquid is prepared by the following method: using crop straw as raw material, heating to 180-220°C under anaerobic conditions, pyrolyzing for 1.5-2.5h, and dissolving the flue gas generated during the pyrolysis process in water by eluting to obtain the tobacco liquid.

[0013] Further preferably, the anoxic conditions are that the oxygen content is ≤12%, the heating rate is 2-4°C / min, and the mass ratio of crop straw to water is 1:1.

[0014] Preferably, the Bacillus amyloliquefaciens dilution solution is obtained by diluting the Bacillus amyloliquefaciens stock solution 20 times with water.

[0015] Preferably, the preparation method of the Bacillus amyloliquefaciens stock solution is: inoculating Bacillus amyloliquefaciens SNMB1 into a fermentation medium, and fermenting and culturing at 28-32° C. for 2-4 days to obtain the Bacillus amyloliquefaciens stock solution.

[0016] Further preferably, the deposit number of Bacillus amyloliquefaciens SNMB1 is CGMCC No.25566.

[0017] Further preferably, the fermentation medium can be a liquid PDA medium or a corn straw medium; the composition of the liquid PDA medium is: 200.0 g potato, 20.0 g glucose, and 1.0 L water; the composition of the corn straw medium is: 37.5 g corn straw powder and 1.0 L water.

[0018] More preferably, the number of viable bacteria in the Bacillus amyloliquefaciens stock solution is ≥ 1×10 7 cfu / mL.

[0019] Preferably, in step (1), in the mixed solution, the volume ratio of the tobacco water dilution solution to the Bacillus amyloliquefaciens dilution solution is 1:1.

[0020] Preferably, in step (1), the root immersion treatment time is 1 hour.

[0021] Preferably, in step (2), the Salvia miltiorrhiza seedlings are transplanted to the field on April 20.

[0022] Preferably, in step (2), the growth and development period of Salvia miltiorrhiza is from June to October every year after transplanting.

[0023] Preferably, in step (2), the spraying treatment operation of the foliage of Salvia miltiorrhiza is as follows: during the growth and development period of Salvia miltiorrhiza, the leaves of Salvia miltiorrhiza are sprayed with the mixed solution once on the 5th of each month.

[0024] Preferably, in step (2), the spraying amount of the mixed solution is 40-50 L / mu each time the spraying treatment is performed.

[0025] Beneficial effects of the present invention:

[0026] The invention dilutes tobacco water and bacillus amyloliquefaciens respectively and then mixes them to obtain a mixed solution, then uses the mixed solution to perform root immersion treatment on salvia miltiorrhiza seedlings, immediately transplants the salvia miltiorrhiza seedlings to a field after the root immersion is completed, and sprays the leaves of the salvia miltiorrhiza with the mixed solution during the growth and development period of the salvia miltiorrhiza seedlings.

[0027] The present invention uses tobacco water and Bacillus amyloliquefaciens to treat Salvia miltiorrhiza by root soaking and foliar spraying, which is beneficial to improve the yield and quality of Salvia miltiorrhiza. Specifically, compared with the untreated group, the method of the present invention can increase the SPAD value and net photosynthetic rate of Salvia miltiorrhiza root expansion period by 19.40% and 9.90%, respectively, and can also increase the accumulation of nitrogen, phosphorus and potassium elements in the Salvia miltiorrhiza root. After transplanting for 180 days, the nitrogen accumulation in the root increased by 62.88%, the phosphorus accumulation increased by 20.75%, and the potassium accumulation increased by 18.03%. At the same time, compared with the untreated group, the single plant accumulation of tanshinone I increased by 20.33%, the single plant accumulation of tanshinone IIA increased by 12.92%, the single plant accumulation of cryptotanshinone increased by 26.53%, and the single plant accumulation of salvianolic acid B increased by 102.01%. The tobacco water and Bacillus amyloliquefaciens have a synergistic effect in increasing the content of the active ingredients in Salvia miltiorrhiza. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 :(A) Effects of different treatments on root fresh weight;(B) Effects of different treatments on root dry weight;

[0029] Figure 2 :(A) Effects of different treatments on dry root weight per plant;(B) Effects of different treatments on yield per hectare;

[0030] Figure 3 :(A) Effects of different treatment groups on leaf color value SPAD during flowering period; (B) Effects of different treatment groups on leaf color value SPAD during root expansion period. DETAILED DESCRIPTION

[0031] It should be noted that the following detailed descriptions are illustrative and are intended to provide further explanation of the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which the present application belongs.

[0032] In order to enable those skilled in the art to more clearly understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below in conjunction with specific embodiments.

[0033] The experimental materials used in the embodiments of the present invention are all conventional experimental materials in the art and can be purchased through commercial channels.

[0034] Embodiment 1:

[0035] (1) placing corn cobs in a pyrolysis furnace, heating the temperature to 200° C. at a heating rate of 3° C. / min under anoxic conditions (oxygen content ≤ 12%, volume fraction), and maintaining the temperature for pyrolysis for 2 hours, collecting the flue gas generated during the pyrolysis process, and dissolving the flue gas in water by eluting to obtain a flue gas water stock solution, wherein the mass ratio of corn cobs to water is 1:1, and diluting the flue gas water stock solution by 500 times with water to obtain a flue gas water dilution solution;

[0036] Bacillus amyloliquefaciens SNMB1 was inoculated into a fermentation medium and fermented at 30°C for 3 days to obtain a Bacillus amyloliquefaciens stock solution; the fermentation medium was a PDA medium, which consisted of 200.0 g of potato, 20.0 g of glucose, and 1.0 L of water. The viable count of the Bacillus amyloliquefaciens stock solution was adjusted to 1×10 7 cfu / mL, the Bacillus amyloliquefaciens stock solution was diluted 20 times with water to obtain a Bacillus amyloliquefaciens dilution solution;

[0037] The diluted tobacco water solution and the diluted Bacillus amyloliquefaciens solution were mixed in a volume ratio of 1:1 to obtain a mixed solution, and the mixed solution was used to soak the roots of Salvia miltiorrhiza seedlings for 1 hour;

[0038] (2) The root-soaked Salvia miltiorrhiza seedlings were transplanted to the field on April 20, and a large ridge double-row planting pattern was adopted, with a ridge width of 80 cm, a ridge surface width of 40 cm, a row spacing of 20 cm, and a plant spacing of 20 cm. When Salvia miltiorrhiza entered the growth and development period from June to October, the mixed solution was sprayed on the leaves of Salvia miltiorrhiza on the 5th of each month during the growth and development period. Each time the mixed solution was sprayed, the spraying amount of the mixed solution was 45 L / mu.

[0039] Embodiment 2:

[0040] (1) placing corn cobs in a pyrolysis furnace, heating the temperature to 220° C. at a heating rate of 2° C. / min under anoxic conditions (oxygen content ≤ 12%, volume fraction), and heat-preserving the temperature for 2.5 hours, collecting the flue gas generated during the heat preservation and pyrolysis process, and dissolving the flue gas in water by eluting to prepare a flue gas water stock solution, wherein the mass ratio of corn cobs to water is 1:1, and diluting the flue gas water stock solution by 500 times with water to obtain a flue gas water dilution solution;

[0041] Bacillus amyloliquefaciens SNMB1 was inoculated into a fermentation medium and fermented at 28°C for 2 days to obtain a Bacillus amyloliquefaciens stock solution; the fermentation medium was a PDA medium, which consisted of 200.0 g of potato, 20.0 g of glucose, and 1.0 L of water. The viable cell count of the Bacillus amyloliquefaciens stock solution was adjusted to 1×10 7 cfu / mL, the Bacillus amyloliquefaciens stock solution was diluted 20 times with water to obtain a Bacillus amyloliquefaciens dilution solution;

[0042] The diluted tobacco water solution and the diluted Bacillus amyloliquefaciens solution were mixed in a volume ratio of 1:1 to obtain a mixed solution, and the mixed solution was used to soak the roots of Salvia miltiorrhiza seedlings for 1 hour;

[0043] (2) After the root-immersion treatment, the Salvia miltiorrhiza seedlings were transplanted to the field on April 20, and a large ridge double-row planting pattern was adopted, with a ridge width of 80 cm, a ridge surface width of 40 cm, a row spacing of 20 cm, and a plant spacing of 20 cm. When Salvia miltiorrhiza entered the growth and development period from June to October, the mixed solution was sprayed on the leaves of Salvia miltiorrhiza on the 5th of each month during the growth and development period. Each time the mixed solution was sprayed, the spraying amount of the mixed solution was 40 L / mu.

[0044] Embodiment 3:

[0045] (1) placing corn cobs in a pyrolysis furnace, heating the temperature to 180° C. at a heating rate of 2-4° C. / min under anoxic conditions (oxygen content ≤ 12%, volume fraction), and heat-preserving the temperature for 1.5 hours, collecting the flue gas generated during the heat preservation and pyrolysis process, and dissolving the flue gas in water by eluting to prepare a flue gas water stock solution, wherein the mass ratio of corn cobs to water is 1:1, and diluting the flue gas water stock solution by 500 times with water to obtain a flue gas water dilution solution;

[0046] Bacillus amyloliquefaciens SNMB1 was inoculated into a fermentation medium and fermented at 32°C for 4 days to obtain a Bacillus amyloliquefaciens stock solution; the fermentation medium was a PDA medium, which consisted of 200.0 g of potato, 20.0 g of glucose, and 1.0 L of water. The viable count of the Bacillus amyloliquefaciens stock solution was adjusted to 1×10 7 cfu / mL, the Bacillus amyloliquefaciens stock solution was diluted 20 times with water to obtain a Bacillus amyloliquefaciens dilution solution;

[0047] The diluted tobacco water solution and the diluted Bacillus amyloliquefaciens solution were mixed in a volume ratio of 1:1 to obtain a mixed solution, and the mixed solution was used to soak the roots of Salvia miltiorrhiza seedlings for 1 hour;

[0048] (2) After the root-immersion treatment, the Salvia miltiorrhiza seedlings were transplanted to the field on April 20, and a large ridge double-row planting pattern was adopted, with a ridge width of 80 cm, a ridge surface width of 40 cm, a row spacing of 20 cm, and a plant spacing of 20 cm. When Salvia miltiorrhiza entered the growth and development period from June to October, the mixed solution was sprayed on the leaves of Salvia miltiorrhiza on the 5th of each month during the growth and development period. Each time the mixed solution was sprayed, the spraying amount was 50 L / mu.

[0049] Test Example 1: Content of medicinal ingredients in Salvia miltiorrhiza

[0050] In order to verify the promoting effect of the method of the present invention on the content of the active ingredients of Danshen, a field experiment was conducted. This experiment set up 5 treatment groups, each of which selected 3 Danshen plants, and the active ingredients of Danshen in each treatment group were detected, including tanshinone I, tanshinone IIA, salvianolic acid B and cryptotanshinone. The specific experimental steps are:

[0051] (1) Purple-flowered Salvia miltiorrhiza was selected as the experimental Salvia miltiorrhiza. Different solutions were used in each treatment group to soak the roots of Salvia miltiorrhiza seedlings for 1 hour. The Salvia miltiorrhiza seedlings after soaking were transplanted to the fields of Mazhuang Town, Daiyue District, Tai'an City, Shandong Province on the same day (April 20). The large ridge double-row planting mode was adopted, with a ridge width of 80 cm, a ridge surface width of 40 cm, a row spacing of 20 cm, and a plant spacing of 20 cm. When Salvia miltiorrhiza entered the growth and development period from June to October, different solutions were used to spray the leaves of Salvia miltiorrhiza on the 5th of each month during the growth and development period. The spraying amount of the mixed solution during the spraying process was 45 L / mu. Salvia miltiorrhiza was harvested on November 5th.

[0052] (2) Take the roots of Salvia miltiorrhiza from each treatment group, dry them, grind them and sieve them, accurately weigh 0.50 g of Salvia miltiorrhiza powder, repeat each treatment 3 times, place it in a 50 mL conical flask, add 35 mL of 60% methanol and perform ultrasonic-assisted extraction for 40 min. The treated sample solution is filtered with a syringe without a needle and a 0.22 μm organic filter membrane and placed in a brown injection vial, and stored at 4°C for later use.

[0053] (3) The results are shown in Table 1. Liquid phase determination conditions: chromatographic column: HolaC18 column, gradient elution 10 min, flow rate 0.3 mL / min, detection wavelength 275 nm, column temperature 35 °C, injection volume 3 μL. Mass spectrometry conditions: positive and negative ion mode, full scan mode (FullMS), scan range set to 133-2000 m / z, resolution 35000;

[0054] Record the chromatogram and draw a regression equation to calculate the relative content of each active ingredient, and use the relative content to calculate the single plant accumulation of each active ingredient. The regression equations corresponding to the relative content of each active ingredient are:

[0055] Salvianolic acid B: y = 3 × 10 6 x-2×10 8 R 2 =0.9988;

[0056] Cryptotanshinone: y = 11157679914.07090x + 119957035.44576R 2 =0.99325;

[0057] Tanshinone I: y = 4063848396.50146x + 35524781.34111R 2 =0.99997;

[0058] Tanshinone IIA: y = 9423402617.39800x + 290916089.29946R 2 =0.99950;

[0059] The calculation formula for the accumulation of a single plant is:

[0060] Accumulation amount per plant (mg / plant) = relative content (mg / g) × root dry weight (g / plant).

[0061] The details of each treatment group are as follows:

[0062] Treatment group 1: no treatment control (CK), this treatment group did not use any solution to soak the roots of Salvia miltiorrhiza and spray them;

[0063] Treatment group 2: water treatment control (W), this treatment group used water to soak the roots of Salvia miltiorrhiza and spray it;

[0064] Treatment group 3: tobacco water treatment (SW), in this treatment group, the tobacco water dilution was used to soak the roots of Salvia miltiorrhiza and spray them, wherein the preparation method of the tobacco water dilution was the same as that of Example 1;

[0065] Treatment group 4: Bacillus amyloliquefaciens treatment (BW), in this treatment group, the roots of Salvia miltiorrhiza were soaked and sprayed with the diluted solution of Bacillus amyloliquefaciens, wherein the preparation method of the diluted solution of Bacillus amyloliquefaciens was the same as that of Example 1;

[0066] Treatment group 5: treatment with a mixed solution of tobacco water and Bacillus amyloliquefaciens (SWBA). In this treatment group, a mixture of tobacco water dilution and Bacillus amyloliquefaciens dilution in a volume ratio of 1:1 was used to soak the roots of Salvia miltiorrhiza and spray them. The preparation method of the tobacco water dilution and Bacillus amyloliquefaciens dilution used in this treatment group was the same as that in Example 1.

[0067] Ensure that the daily management of each treatment group is consistent during the experiment.

[0068] Table 1 Effects of tobacco water and Bacillus amyloliquefaciens mixed solution on the active ingredients of Salvia miltiorrhiza

[0069]

[0070]

[0071] As can be seen from Table 1, the treatment with the mixed solution of tobacco water and Bacillus amyloliquefaciens (SWBA) significantly increased the contents of tanshinone I, tanshinone IIA, cryptotanshinone and salvianolic acid B in Salvia miltiorrhiza. Specifically, compared with the untreated control (CK) and the water-treated control (W), the single-plant accumulation of tanshinone I increased by 20.33% and 22.95%, the single-plant accumulation of tanshinone IIA increased by 12.92% and 14.19%, the single-plant accumulation of cryptotanshinone increased by 26.53% and 28.85%, and the single-plant accumulation of salvianolic acid B increased by 102.01% and 97.79%, respectively.

[0072] At the same time, in terms of cryptotanshinone content, compared with the untreated group, the single-plant accumulation of cryptotanshinone in Danshen increased by 7.86 mg / plant, 7.05 mg / plant and 19.59 mg / plant, respectively, in the treatment of tobacco water alone, Bacillus amyloliquefaciens alone, and tobacco water and Bacillus amyloliquefaciens combination. In terms of salvianolic acid B content, compared with the untreated group, the single-plant accumulation of salvianolic acid B in Danshen increased by 804.95 mg / plant, 1465.38 mg / plant and 2855.89 mg / plant, respectively, in the treatment of tobacco water alone, Bacillus amyloliquefaciens alone, and tobacco water and Bacillus amyloliquefaciens combination. It can be seen that the mixed solution obtained by the combination of tobacco water and Bacillus amyloliquefaciens is used for root soaking and spraying of Danshen, which has a synergistic effect in increasing the content of active ingredients in Danshen, especially in increasing the content of salvianolic acid B and cryptotanshinone.

[0073] Test Example 2: Agronomic traits and yield of Salvia miltiorrhiza

[0074] In order to verify the promoting effect of the method of the present invention on the quality and yield of Salvia miltiorrhiza, a field experiment was conducted. This experiment set up 3 treatment groups, and the root fresh weight, single plant root dry weight, unit area yield, SPAD, net photosynthetic rate, stomatal conductance, root diatom, phosphorus and potassium accumulation of Salvia miltiorrhiza in each treatment group were detected respectively. The specific experimental steps are:

[0075] Purple flowered Salvia miltiorrhiza was selected as the experimental Salvia miltiorrhiza. Different solutions were used in each treatment group to soak the roots of Salvia miltiorrhiza seedlings for 1 hour. The Salvia miltiorrhiza seedlings after soaking were transplanted to the fields of Mazhuang Town, Daiyue District, Tai'an City, Shandong Province on the same day (April 20). The large ridge double-row planting mode was adopted, with ridge width of 80cm, ridge surface width of 40cm, row spacing of 20cm, and plant spacing of 20cm. When Salvia miltiorrhiza entered the growth and development period from June to October, different solutions were used to spray the leaves of Salvia miltiorrhiza on the 5th of each month during the growth and development period. The spraying amount of the mixed solution during the spraying process was 45L / mu. Salvia miltiorrhiza was harvested on November 5, and the root fresh weight, single plant root dry weight, unit area yield, SPAD, net photosynthetic rate, stomatal conductance, root diatom nitrogen, phosphorus and potassium accumulation of Salvia miltiorrhiza were tested.

[0076] The details of each treatment group are as follows:

[0077] Treatment group 1: no treatment control (CK), this treatment group did not use any solution to soak the roots of Salvia miltiorrhiza and spray them;

[0078] Treatment group 2: water treatment control (W), this treatment group used water to soak the roots of Salvia miltiorrhiza and spray it;

[0079] Treatment group 3: treatment with a mixed solution of tobacco water and Bacillus amyloliquefaciens (SWBA). In this treatment group, a mixture of tobacco water dilution and Bacillus amyloliquefaciens dilution in a volume ratio of 1:1 was used to soak the roots of Salvia miltiorrhiza and spray them. The preparation method of the tobacco water dilution and Bacillus amyloliquefaciens dilution used in this treatment group was the same as that in Example 1.

[0080] Ensure that the daily management of each treatment group is consistent during the experiment.

[0081] (1) Root fresh weight and root dry weight

[0082] Field plant sampling began on the 60th day after transplanting. Five Salvia miltiorrhiza plants with uniform growth were randomly selected from each treatment. Sampling was carried out on the same day of each month thereafter, for a total of five times. The root fresh weight of the samples after sampling was measured, and then the fresh root samples were dried at 60°C and weighed to obtain the root dry weight after constant weight. The results are as follows: Figure 1 shown.

[0083] Depend on Figure 1 It can be seen that 180 days after transplanting, in terms of root dry weight, the root dry weight of the treatment with the mixed solution of tobacco water and Bacillus amyloliquefaciens (SWBA) increased by 26.84% compared with the untreated control (CK) and 31.07% compared with the water-treated control (W). In terms of root fresh weight, the root fresh weight of the treatment with the mixed solution of tobacco water and Bacillus amyloliquefaciens (SWBA) increased by 3.50% compared with the untreated control (CK) and 16.16% compared with the water-treated control (W). It can be seen that the treatment with the mixed solution of tobacco water and Bacillus amyloliquefaciens (SWBA) significantly increased the root dry weight.

[0084] (2) Dry weight of single root and yield per unit area

[0085] After harvesting Salvia miltiorrhiza on November 5, random sampling of 2m×0.8m sampling areas was carried out in the yield measurement area corresponding to each treatment group, and 20 plants were selected from each sampling area. Ten representative plants were selected from each treatment group, air-dried to constant weight under natural conditions, and the dry weight of the roots was weighed to calculate the dry weight of the root of each plant.

[0086] At the same time, the root dry weight of 20 plants in the sampling area was weighed to calculate the yield per unit area. The calculation formula of the yield per unit area is shown in formula (I):

[0087]

[0088] Depend on Figure 2 It can be seen that in terms of single root dry weight, the Salvia miltiorrhiza treated with the tobacco water and Bacillus amyloliquefaciens complex mixture (SWBA) increased by 10.41% and 5.35% compared to the untreated control (CK) and the water-treated control (W), respectively. In terms of yield per unit area, the tobacco water and Bacillus amyloliquefaciens complex mixture (SWBA) treatment increased by 10.14% compared to the untreated control (CK) and increased by 5.56% compared to the water-treated control (W). It can be seen that the present invention can significantly increase the single root dry weight and unit area yield of Salvia miltiorrhiza by using straw-source tobacco water and Bacillus amyloliquefaciens complex.

[0089] (3) Leaf color value SPAD

[0090] During the flowering period (July 20) and root expansion period (September 20) of Salvia miltiorrhiza, the leaf SPAD value was measured using a portable SPAD-502Plus chlorophyll meter. It was calibrated before use. Ten Salvia miltiorrhiza plants with consistent growth were selected from each treatment group, and the middle of healthy leaves with consistent growth positions were measured. The results were repeated 3 times. Figure 3 shown.

[0091] Depend on Figure 3 It can be seen that during the flowering period, the leaf color value SPAD after the treatment with the mixed solution of tobacco water and Bacillus amyloliquefaciens (SWBA) was 25.28% higher than that of the untreated control (CK) and 20.56% higher than that of the water-treated control (W). During the root expansion period, the leaf color value SPAD after the treatment with the mixed solution of tobacco water and Bacillus amyloliquefaciens (SWBA) was 19.40% higher than that of the untreated control (CK) and 15.22% higher than that of the water-treated control (W). It can be seen that the present invention can significantly improve the leaf color value of Salvia miltiorrhiza during the flowering period and the root expansion period by using the straw-source tobacco water and Bacillus amyloliquefaciens compound.

[0092] (4) Photosynthetic related parameters

[0093] During the flowering period (July 20) and root expansion period (September 20) of Salvia miltiorrhiza, photosynthesis-related parameters were measured using a LI-6400XT portable photosynthesis meter from 9:00 to 11:30 in the morning on a clear day. Fifteen plants were measured in each treatment group, with three replicates per plant. The photosynthesis-related parameters included net photosynthetic rate (Pn), transpiration rate (Tr), intercellular CO2 concentration (Ci), and stomatal conductance (Gs). The results are shown in Table 2.

[0094] Table 2 Effects of tobacco water and Bacillus amyloliquefaciens mixed solution on photosynthesis-related parameters

[0095]

[0096] As can be seen from Table 2, in the flowering stage, the net photosynthetic rate of the straw-derived smoke water and Bacillus amyloliquefaciens mixed solution (SWBA) was increased by 17.47% and 16.45% respectively compared with the untreated control (CK) and the water-treated control (W). In the root swelling stage, the net photosynthetic rate of the straw-derived smoke water and Bacillus amyloliquefaciens mixed solution was increased by 9.90% compared with the untreated control (CK) and the water-treated control (W). It can be seen that the treatment of Salvia miltiorrhiza with the mixed solution of straw-derived smoke water and Bacillus amyloliquefaciens can increase the net photosynthetic rate.

[0097] (5) Accumulation of nitrogen, phosphorus and potassium

[0098] The root, stem and leaf parts of the Salvia miltiorrhiza plant were boiled by H2SO4-H2O2 digestion method. The Salvia miltiorrhiza plant was used as the sample to be tested, and each treatment was repeated 3 times. The specific operation is:

[0099] Weigh 0.20g of Salvia miltiorrhiza plant and add it to a digestion tube. After adding a little distilled water to moisten it, add 5mL of concentrated sulfuric acid, place it on a 340℃ high-temperature digestion furnace, quickly add 1 dropper of 30% H2O2, digest for 20 minutes, then add 2 droppers of 30% H2O2 and boil for 30 minutes, until the solution becomes transparent and colorless. After the digestion liquid is naturally cooled, the volume is adjusted to 50mL as the test solution. After 2mL of the test solution is drawn and added to 3mL of deionized water to mix, the total nitrogen and total phosphorus are determined by a flow analyzer (AA3, SEAL) and the mass fractions of nitrogen and phosphorus are calculated. The results are shown in Tables 3-4. Among them, the mass fraction calculation formula of nitrogen is shown in formula (II), and the mass fraction calculation formula of phosphorus is shown in formula (III).

[0100]

[0101] In formula (II), ω(N) is the mass fraction of nitrogen, mg·kg -1 ; c is the flow analyzer test result, mg·L -1 ; m is the mass of the sample to be tested, g;

[0102]

[0103] In formula (III), ω(P) is the mass fraction of phosphorus, mg·kg -1 ;c P is the test result of flow analyzer, mg·L -1 ; m is the mass of the sample to be tested, g.

[0104] The potassium content was determined by flame photometry. The specific operation is as follows:

[0105] Accurately weigh 1.907g KCl (pre-dried at 110℃ for 2h), dilute to 1000mL to obtain the first standard solution, accurately pipette 10mL of the first standard solution, add water and dilute to 100mL to obtain the second standard solution, respectively pipette 0mL, 5mL, 10mL, 20mL, 40mL, 60mL, and 80mL of the second standard solution into a 100mL volumetric flask, add 0.1mL of concentrated sulfuric acid, dilute to volume with deionized water to obtain 0, 5, 10, 20, 40, 60, and 80mg / L K standard solutions, measure the flame photometry value of each K standard solution, draw a standard curve, and calculate the potassium content. The results are shown in Table 5.

[0106] The calculation formula of potassium content is shown in formula (IV):

[0107]

[0108] In formula (IV), ω(K) is the mass fraction of potassium, mg·kg -1 ; c2 is the potassium concentration in the sample solution obtained from the standard curve, μg / mL; c1 is the potassium concentration in the sample blank solution obtained from the standard curve, μg / mL; v is the volume of the sample solution, mL; n is the dilution factor; m is the mass of the sample, g.

[0109] The nitrogen, phosphorus and potassium contents of a single plant were calculated using the mass fractions of nitrogen and phosphorus and the potassium content calculated above. The calculation formula is shown in formula (V).

[0110] Nitrogen / phosphorus / potassium content of a single plant = ω(N / P / K)×50×(0.001 / 0.2)×root dry weight (V).

[0111] Table 3 Effect of the mixed solution of tobacco water and Bacillus amyloliquefaciens on nitrogen accumulation of Salvia miltiorrhiza (mg / plant)

[0112]

[0113]

[0114] As can be seen from Table 3, 180 days after transplanting, the increases in nitrogen accumulation in the roots were 62.88% and 34.29% compared with the untreated control (CK) and the water-treated control (W), respectively. This shows that soaking and spraying the roots of Salvia miltiorrhiza with the mixed solution obtained by mixing straw smoke water and Bacillus amyloliquefaciens can increase the nitrogen accumulation in Salvia miltiorrhiza.

[0115] Table 4 Effect of the mixed solution of tobacco water and Bacillus amyloliquefaciens on phosphorus accumulation in Salvia miltiorrhiza (mg / plant)

[0116]

[0117] As can be seen from Table 4, 180 days after transplanting, the treatment with smoke water and Bacillus amyloliquefaciens mixed solution (SWBA) significantly increased the phosphorus content in the roots of Salvia miltiorrhiza. Compared with the untreated control (CK) and water treatment control (W), the increases in phosphorus accumulation in the roots were 20.75% and 10.73%, respectively.

[0118] Table 5 Effect of tobacco water and Bacillus amyloliquefaciens mixed solution on potassium accumulation in Salvia miltiorrhiza (mg / plant)

[0119]

[0120] As can be seen from Table 5, 180 days after transplanting, the treatment with smoke water and Bacillus amyloliquefaciens mixed solution (SWBA) significantly increased the potassium content in the roots of Salvia miltiorrhiza. Compared with the untreated control (CK) and water treatment control (W), the increases in potassium accumulation in the roots were 18.03% and 39.28%, respectively.

[0121] The above description is only the preferred 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 modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A method for improving the quality and increasing the yield of Salvia miltiorrhiza based on straw-derived tobacco water and Bacillus amyloliquefaciens, characterized in that: The following steps are involved: (1) mixing the diluted tobacco water solution and the diluted Bacillus amyloliquefaciens solution to obtain a mixed solution, and using the mixed solution to perform root immersion treatment on Salvia miltiorrhiza seedlings; (2) After the root-immersion treatment of the Salvia miltiorrhiza seedlings is transplanted to the field, the mixed solution is sprayed on the leaves of the Salvia miltiorrhiza during its growth and development period.

2. The method for improving the quality and increasing the yield of Salvia miltiorrhiza according to claim 1, characterized in that: The volume ratio of the tobacco water dilution liquid and the Bacillus amyloliquefaciens dilution liquid in the mixed liquid is 1:1; the tobacco water dilution liquid is obtained by diluting the tobacco water stock liquid with water to 500 times, and the Bacillus amyloliquefaciens dilution liquid is obtained by diluting the Bacillus amyloliquefaciens stock liquid with water to 20 times.

3. The method for improving the quality and increasing the yield of Salvia miltiorrhiza as claimed in claim 2, characterized in that: The smoke water stock solution is prepared by the following method: using crop straw as raw material, heating to 180-220°C under anaerobic conditions, pyrolyzing for 1.5-2.5h, dissolving the smoke generated during the pyrolysis process in water through leaching, and obtaining the smoke water stock solution.

4. The method for improving the quality and increasing the yield of Salvia miltiorrhiza as claimed in claim 3, characterized in that: The anoxic conditions were oxygen content ≤ 12%, a heating rate of 2-4°C / min, and a mass ratio of crop straw to water of 1:

1.

5. The method for improving the quality and increasing the yield of Salvia miltiorrhiza as claimed in claim 2, characterized in that: The number of viable bacteria in the Bacillus amyloliquefaciens stock solution is ≥ 1×10 7 cfu / mL.

6. The method for improving the quality and increasing the yield of Salvia miltiorrhiza according to claim 2, characterized in that: The preparation method of the Bacillus amyloliquefaciens stock solution is as follows: inoculating Bacillus amyloliquefaciens SNMB1 into a fermentation medium, and fermenting and culturing at 28-32° C. for 2-4 days to obtain the Bacillus amyloliquefaciens stock solution.

7. The method for improving the quality and increasing the yield of Salvia miltiorrhiza according to claim 6, characterized in that: The deposit number of Bacillus amyloliquefaciens SNMB1 is CGMCC No.25566.

8. The method for improving the quality and increasing the yield of Salvia miltiorrhiza according to claim 1, characterized in that: In step (1), the root immersion treatment time is 1 hour.

9. The method for improving the quality and increasing the yield of Salvia miltiorrhiza according to claim 1, characterized in that: In step (2), the time for transplanting the Salvia miltiorrhiza seedlings to the field is April 20, and the growth and development period of Salvia miltiorrhiza is from June to October every year.

10. The method for improving the quality and increasing the yield of Salvia miltiorrhiza according to claim 1, characterized in that: In step (2), the spraying amount of the mixed solution is 40-50 L / mu each time the spraying treatment is performed.

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