Endophytic fungus AP12 capable of promoting growth of andrographis paniculata plants and accumulation of active ingredient content and application of endophytic fungus AP12

Abstract

The invention discloses an endophytic fungus AP12 capable of promoting growth of andrographis paniculata plants and accumulation of active ingredient content and application of the endophytic fungus AP12. The colletotrichum sp. AP12 endophytic fungus provided by the invention can promote the growth of a common andrographis herb plant and the accumulation of the content of effective components, and after the colletotrichum sp. AP12 is applied and cultured under a pot culture condition, the leaf area of the common andrographis herb can be obviously increased, the fresh weight and the dry weight can be increased, the accumulation of the content of dry matters can be increased, and the growth of the common andrographis herb plant can be obviously promoted; meanwhile, the accumulation of active ingredients of andrographolide, neoandrographolide, 14-deoxidized andrographolide and dehydroandrographolide of andrographis paniculata can be remarkably promoted, the content of total lactone is increased, and it is shown that the endophytic fungi AP12 are beneficial endophytic fungi capable of promoting growth and metabolism of andrographis paniculata; and reference is provided for application and development of the endophytic fungi in promotion of growth of andrographis paniculata and accumulation of effective component content.

Description

Technical Field

[0001] The present invention belongs to the field of microbial technology, and more specifically relates to an endophytic fungus AP12 capable of promoting the growth of Andrographis paniculata plants and the accumulation of effective components, and its application. Background Art

[0002] Andrographis paniculata is the dried aerial part of the plant Andrographis paniculata (Burm.f.) Nees of the Acanthaceae family. It has the effects of clearing away heat and detoxifying, cooling blood, and reducing swelling. It is used to relieve symptoms such as colds, fever, and sore throats. It is now widely distributed in tropical and subtropical regions of Asia, including China, India, and Thailand. At present, more than 80 chemical components have been isolated and identified from Andrographis paniculata, such as diterpenoid lactones, flavonoids, and polyphenols. Among them, the main medicinal active ingredients are diterpenoid lactone compounds of Andrographis paniculata, most of which are concentrated in the leaves of Andrographis paniculata, including andrographolide, andrographolide, 14-deoxyandrographolide, andrographolide dehydrate, etc. These active ingredients are considered to be "natural antibiotics" with pharmacological effects such as anti-inflammatory, antibacterial, antiviral, and hypoglycemic. Due to the rich medicinal and therapeutic effects of Andrographis paniculata, the market demand for its medicinal materials has increased greatly. However, due to the existence of many unfavorable factors such as single germplasm, environmental damage, and crop rotation obstacles, Andrographis paniculata has caused genetic variation, separation, and degradation, germplasm mixing, and the content and quality of medicinal active ingredients are unstable. Therefore, it is urgent to breed Andrographis paniculata varieties with high yield, stable quality, good medicinal active ingredients, and high content.

[0003] At present, studies have shown that microbial strains have great development potential in promoting plant growth, improving the host's stress resistance and disease resistance, and increasing the accumulation of the host's secondary metabolite content. For example, Chinese patent CN110063161A discloses that the use of nitrogen-fixing rhizobia in Sesbania nodules can promote the growth of Andrographis paniculata and increase its effective ingredient content, and the use of biological nitrogen fixation can promote the growth of interactive plants and the accumulation of andrographolide and dehydroandrographolide. However, the microbial strains that can be used to promote the growth of Andrographis paniculata plants and increase the accumulation of effective ingredient content are very limited. There are no other strains of microbial strains that can promote the growth of Andrographis paniculata plants and the accumulation of effective ingredient content in the prior art. Therefore, the research and development of microbial strains that can promote the accumulation of Andrographis paniculata effective ingredient content is of great significance to the cultivation of excellent Andrographis paniculata plants and the improvement of higher Andrographis paniculata medicinal active ingredients or effective ingredient content. Summary of the invention

[0004] The technical problem to be solved by the present invention is to overcome the defects and shortcomings of the above-mentioned problems and provide an endophytic fungus AP12 which can promote the growth of Andrographis paniculata plants and the accumulation of effective ingredient content and its application.

[0005] The first object of the present invention is to provide an endophytic fungus Colletotrichum sp. AP12 strain that can promote the growth of Andrographis paniculata plants and the accumulation of effective ingredients.

[0006] The second object of the present invention is to provide the application of the Colletotrichum AP12 strain.

[0007] The third object of the present invention is to provide a growth-promoting agent of Andrographis paniculata and / or a preparation having increased content of effective ingredients of Andrographis paniculata.

[0008] The fourth object of the present invention is to provide a method for increasing the content of effective ingredients in Andrographis paniculata.

[0009] The fifth object of the present invention is to provide a method for promoting the growth of Andrographis paniculata.

[0010] The above-mentioned purpose of the present invention is achieved through the following technical solutions:

[0011] The present invention provides an endophytic fungus Colletotrichum sp. AP12 strain that can promote the growth of Andrographis paniculata plants and the accumulation of effective ingredient content. The strain was deposited in the Guangdong Provincial Microbiological Culture Collection Center on June 15, 2023, and the deposit number is GDMCC No: 63546. The AP12 strain provided by the present invention is isolated from the leaf tissue of Andrographis paniculata, and is identified as Colletotrichum sp. by morphological characteristics and phylogenetic analysis. The homology reaches 99%. The middle part of the AP12 bacterial body is convex, the colony is regular and round, the culture medium is dyed orange-yellow, the middle part of the front is dark green, the mycelium surface has gray-black balls, curled, and the edges are orange-red, and the middle part of the back is also dark green, and the edges are orange-red.

[0012] The research of the present invention shows that the endophytic fungus AP12 can significantly promote the growth of Andrographis paniculata plants, increase plant height, leaf number and leaf area, and can increase the chlorophyll content of Andrographis paniculata, increase the fresh weight of the whole plant, the fresh weight of the aboveground part, the dry weight of the whole plant and the dry weight of the aboveground part of the plant, and promote the accumulation of fresh and dry matter content of Andrographis paniculata plants; at the same time, the endophytic fungus strain AP12 can also increase the content of Andrographis paniculata effective ingredients: andrographolide, neoandrographolide, 14-deoxyandrographolide, dehydroandrographolide, significantly increase the content of effective ingredients, and promote the accumulation of its total lactone content, indicating that the strain of Colletotrichum spp. AP12 is an endophytic fungus that is beneficial to the growth and metabolism of Andrographis paniculata plants.

[0013] Therefore, the following applications of the Colletotrichum AP12 strain or its bacterial solution are all within the protection scope of the present invention:

[0014] Application in promoting the growth of Andrographis paniculata.

[0015] Application in increasing the content of active ingredients in Andrographis paniculata.

[0016] Application in the preparation of Andrographis paniculata growth promoter and / or preparation for increasing the content of Andrographis paniculata active ingredients.

[0017] Preferably, the Andrographis paniculata active ingredient is andrographolide.

[0018] More preferably, it is andrographolide, neoandrographolide, 14-deoxyandrographolide and / or dehydroandrographolide.

[0019] The invention provides an Andrographis paniculata growth promoter and / or a preparation for increasing the content of Andrographis paniculata active ingredients, comprising the AP12 strain of Colletotrichum spp. or its bacterial liquid.

[0020] The invention provides a method for improving the content of effective components of Andrographis paniculata. The method comprises adopting a Colletotrichum AP12 strain or a bacterial liquid thereof to process Andrographis paniculata.

[0021] Preferably, the Andrographis paniculata AP12 strain or its bacterial liquid is used to inoculate Andrographis paniculata.

[0022] Preferably, the concentration of the bacterial solution is not less than 30 mg / L, or it is diluted 2 times for use.

[0023] Preferably, the bacterial liquid is fermentation liquid, inactivated fermentation liquid, mycelial liquid or inactivated mycelial liquid.

[0024] The present invention also provides a method for promoting the growth of Andrographis paniculata, which is characterized in that the Andrographis paniculata is treated with the AP12 strain of Colletotrichum spp. or its bacterial liquid.

[0025] The present invention has the following beneficial effects:

[0026] The invention provides an endophytic fungus of Colletotrichum sp. AP12 capable of promoting the growth of andrographolide plants and the accumulation of effective component content. The Colletotrichum sp. AP12 strain is separated and purified from Andrographolide leaf tissue. Studies have shown that after the Colletotrichum sp. AP12 strain is used for planting and culturing under potted conditions, the leaf area of ​​Andrographolide can be significantly increased, the fresh weight and dry weight can be improved, the accumulation of dry matter content can be increased, and the growth of Andrographolide plants can be significantly promoted. At the same time, the accumulation of andrographolide, neoandrographolide, 14-deoxyandrographolide and dehydroandrographolide, which are effective components of Andrographolide, can be significantly promoted, and the content of total lactone can be improved, indicating that the endophytic fungus AP12 strain is a beneficial endophytic fungus capable of promoting the growth and metabolism of Andrographolide.

[0027] The AP12 strain of the genus Colletotrichum provided by the present invention can promote the growth of Andrographis paniculata plants and the accumulation of effective ingredient content, provide a reference for improving the cultivation technology of Andrographis paniculata by using endophytic fungi in the future, further improve the quality and yield of Andrographis paniculata, and provide a reference for the application and development of endophytic fungi in promoting the growth of Andrographis paniculata and the accumulation of effective ingredient content. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is the strain morphology of Colletotrichum AP12 (A is the front of the colony, and B is the back of the colony).

[0029] Figure 2 This is the phylogenetic tree of Colletotrichum AP12.

[0030] Figure 3 This is a picture of the bacterial solution after fermentation of Colletotrichum AP12. DETAILED DESCRIPTION

[0031] The present invention is further described below in conjunction with the accompanying drawings and specific examples, but the examples do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0032] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0033] The formula of each culture medium in the following implementation case is as follows:

[0034] PDA solid culture medium: potato 200 g / L, glucose 20 g / L, agar 14 g / L.

[0035] PDB liquid culture medium: potato 200g / L, glucose 20g / L.

[0036] Example 1 Isolation, purification and molecular identification of endophytic fungi from Andrographis paniculata

[0037] The leaves of Andrographis paniculata were cleaned and disinfected with 75% ethanol for 30 seconds, disinfected with 4% sodium hypochlorite for 15-20 minutes, rinsed with sterile water three times, cut into 0.5mm×0.5mm small cubes and placed in fresh PDA culture medium, and cultured in a 28℃ constant temperature box for about 5-7 days. The sterile water of the last rinse was used as a blank control to verify whether the leaves were thoroughly disinfected; then the edge hyphae were continuously picked and purified on a new PDA culture medium until a single strain was obtained, which was named AP12. Two portions of the obtained single strain AP12 hyphae were preserved in 30% glycerol at 4℃, and the rest were preserved using the slant PDA plate method.

[0038] Fungal genomic DNA was extracted by CTAB method, and DNA concentration and purity were detected by ultra-micro spectrophotometer. PCR products were amplified using universal upper and lower primers ITS1 and ITS4 (ITS1: 5'-TCCGTAGGTGAACCTGCGG-3' and ITS4: 5'-TCCTCCGCTTATTGATATGC-3'). The 50 μL PCR system consisted of: Master mix (green enzyme) 25 μL, primers (diluted to 10 μmol·L -1 ) 2 μL each of ITS1 and ITS4, 1 μL template DNA, add sterile ddH 2 0 to 20 μL, mix well and centrifuge. PCR conditions: 95℃ pre-denaturation for 5 min; 94℃ denaturation for 30 min, 55℃ annealing for 30 s, 72℃ extension for 45 s, 35 cycles; final extension at 72℃ for 10 min, incubation at 4℃. PCR products were detected by agarose gel electrophoresis (electrophoresis conditions were 100 V, 45 min).

[0039] After preliminary inspection, qualified DNA samples were sent to a sequencing company (Shanghai Bioengineering) for sequencing. The sequencing results were uploaded to the website of the National Bioinformatics Institute (https: / / blast..Ncbi.nlm.nih.gov / Blast.cgi) for sequence alignment, and the phylogenetic tree was constructed using the neighbor-joining method using MEGA7.0.26 software.

[0040] The strain morphology of the endophytic fungus AP12 from Andrographis paniculata is as follows Figure 1 As shown, the middle part of the bacterial body is convex, the colony is regular and round, the culture medium is stained orange, and the front side ( Figure 1 A) The middle part is dark green, with gray-black balls on the surface of the hyphae, curled, with orange-red edges, and the back ( Figure 1 B) The middle part is also dark green, with orange-red edges.

[0041] The sequencing comparison results are as follows Figure 2 As shown, the strain AP12 is most closely related to Colletotrichum sp., with 100% and 99% homology, and combined with the colony morphology observation of strain AP12 and the homology analysis in the DNA phylogenetic tree, the endophytic fungus AP12 of Andrographis paniculata was finally identified as Colletotrichum sp. Therefore, the strain isolated and purified by the present invention was identified as the endophytic fungus Colletotrichum sp. of Andrographis paniculata, named as strain AP12, which was deposited in Guangdong Provincial Microbiological Culture Collection Center on June 15, 2023, with a deposit number of GDMCCNo: 63546, and a deposit address of 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou.

[0042] Example 2 Fermentation of endophytic fungus AP12 and preparation of its elicitor

[0043] The endophytic fungus Colletotrichum sp. AP12 strain isolated and purified in Example 1 was used. Eight fungus pieces were taken out on PDA solid medium using a sterile hole puncher with a diameter of 0.8 cm and placed on 400 mL PDB liquid medium. The pieces were fermented in a constant temperature incubator at 28° C. and a rotation speed of 150 r / min for 10 days. The fermented endophytic fungus AP12 was as follows: Figure 3 As shown, the overall bacterial liquid is dark green.

[0044] The fermented mycelium and the fermentation liquid are separated. One part of the fermentation liquid is sterilized at 121°C for 20 minutes by high pressure sterilization to prepare an inactivated fermentation liquid (IFS) with a concentration of 2 times the diluted dose; the other part is not sterilized to prepare a fermentation liquid (FS) with a concentration of 2 times the diluted dose.

[0045] After the mycelium was rinsed with sterile water for several times until no culture medium remained, it was dried with sterile filter paper, 2.0 g of mycelium was weighed and fully ground, mixed evenly with 200 mL of sterile water, and ultrasonically treated for 30 minutes to obtain the mycelium solution to be used, and the polysaccharide concentration in the mycelium solution was detected by phenol-sulfuric acid method with glucose as the standard, a part of it was inactivated to prepare 30 mg / L inactivated mycelium solution (Inactivated Mycelium Solution, referred to as IMS); a part of it was not inactivated to prepare 30 mg / L mycelium solution (Mycelium Solution, referred to as MS). The IFS, FS, IMS and MS are all 4 inducer components of the genus AP12.

[0046] Example 3: Soil culture potting test of Andrographis paniculata plants with AP12 fungal elicitor

[0047] Peat soil: vermiculite: perlite were mixed in a ratio of 2:1:0.5, and sterilized at 121°C for 1h to prepare sterile soil for later use. The seeds of Andrographis paniculata with full grains and uniform quality were preferably sown in sterile soil, and the amount of soil per pot was strictly controlled. The four AP12 fungal elicitor components screened and prepared in Example 2 were used for seeding once every 3 days, 20 mL each time, and the control group was treated with an equal amount of sterile water, 3 plants per group, and each treatment was repeated 12 times. The plants were cultured at 26±2°C, 60% humidity, and 12h alternating light and dark conditions. The growth indicators (plant height, number of leaves, specific leaf area), physiological indicators (chlorophyll content, nitrogen content), and biomass indicators (whole plant fresh weight, aboveground fresh weight, whole plant dry weight, aboveground dry weight) of Andrographis paniculata plants were detected on the 65th day.

[0048] Growth index detection: Use a ruler to measure the height of Andrographis paniculata plants, and use a living leaf area measuring instrument to detect the leaf area.

[0049] Physiological index detection: The chlorophyll content of four endophytic fungal AP12 inducer components (IFS, FS, IMS and MS) and sterile water-irrigated Andrographis paniculata plants cultured in the greenhouse for 65 days was measured using a chlorophyll meter. Each measurement was repeated 3 times and the average value was taken.

[0050] Biomass index detection: Wash the plant with distilled water, dry it with absorbent paper, accurately weigh the fresh weight of the whole plant of Andrographis paniculata, remove the roots of Andrographis paniculata and weigh the fresh weight of the aboveground part, further dry the plant in an oven at 45°C and weigh the dry weight of the whole plant of Andrographis paniculata, and similarly remove the roots and weigh the dry weight of the aboveground part. Repeat each measurement 3 times and take the average value.

[0051] The growth index detection results are shown in Table 1. The four fungal inducer components of the genus Colletotrichum AP12 can all promote the growth of Andrographis paniculata plants. The plant height, number of leaves and leaf area are all higher than those of the control group. The induction effect of the mycelium part is more significant than that of the fermentation liquid part. Among them, the induction effect of the IMS inducer component is the best, especially in terms of leaf area. The application of IMS component and MS component can significantly increase the leaf area of ​​Andrographis paniculata.

[0052] Table 1 Effects of endophytic fungus AP12 on the growth of Andrographis paniculata plants after 65 days of inoculation (n=3)

[0053]

[0054] Note: CK: blank group, IFS: sterilized fermentation liquid; FS: fermentation liquid; IMS: sterilized mycelial liquid; MS: mycelial liquid. Different lowercase letters in the same column represent significant differences (p < 0.05) (statistically significant).

[0055] The results of physiological index detection are shown in Table 2 below. The four fungal inducer components of Colletotrichum AP12 can all promote the growth of Andrographis paniculata plants. Among them, the chlorophyll content and nitrogen content of the IMS component were higher than those of the blank group, and there were significant differences (p < 0.05), indicating that Colletotrichum AP12 can increase the chlorophyll content of Andrographis paniculata and provide energy and substances for various metabolic activities.

[0056] Table 2 Effects of endophytic fungus AP12 on the physiology of Andrographis paniculata plants after 65 days of inoculation (n=3)

[0057]

[0058]

[0059] Note: CK: blank group, IFS: sterilized fermentation liquid; FS: fermentation liquid; IMS: sterilized mycelial liquid; MS: mycelial liquid. Different lowercase letters in the same column represent significant differences (p < 0.05) (statistically significant).

[0060] The results of biomass index detection are shown in Table 3 below. The four fungal elicitor components of the genus Colletotrichum AP12 can promote the accumulation of fresh and dry matter content in Andrographis paniculata plants. Among them, the promotion effect of the mycelium part is better than that of the fermentation liquid part, and the IMS component has the most significant effect on the accumulation of fresh and dry matter content in Andrographis paniculata; the values ​​of the whole plant fresh weight, the aboveground fresh weight, the whole plant dry weight and the aboveground dry weight all reached the highest, and there were significant differences (p < 0.05).

[0061] Table 3 Effects of endophytic fungus AP12 on biomass indexes of Andrographis paniculata plants after 65 days of inoculation (n=3)

[0062]

[0063] Note: CK: blank group, IFS: sterilized fermentation liquid; FS: fermentation liquid; IMS: sterilized mycelial liquid; MS: mycelial liquid. Different lowercase letters in the same column represent significant differences (p < 0.05) (statistically significant).

[0064] Example 4 Detection of the content of active ingredients after the AP12 fungal elicitor was planted in Andrographis paniculata plants

[0065] The Andrographis paniculata plants of the 65th day were collected from the four AP12 fungal elicitor treatment groups and the control group in Example 3, and the contents of effective ingredients (andrographolide, neoandrographolide, 14-deoxyandrographolide and dehydroandrographolide) in the Andrographis paniculata plants were detected.

[0066] The preparation of andrographolide standards and test products refers to the method of the 2020 edition of the "Chinese Pharmacopoeia" (Chinese Pharmacopoeia Committee, 2020 edition). After the sample is simply cleaned, it is dried in an oven at 45°C; the liquid nitrogen grinding method is used to pass the ground powder through a No. 4 sieve, and then 0.100±0.002g of powder is accurately weighed, soaked in 5mL of 40% methanol for 1h, weighed, ultrasonically treated for 30min (power 250W, frequency 33KHz), cooled, weighed again, and 40% methanol was used to make up the lost weight. Shake well, centrifuge at 10000r / min at 4°C for 2min, and the supernatant is passed through a 0.22μm microporous filter membrane to obtain a 20mg / mL test solution.

[0067] Accurately weigh the andrographolide standard, add methanol to make a solution containing 0.302 mg of andrographolide per 1 mL, and filter through a 0.22 μm microporous filter membrane to obtain the standard solution.

[0068] After detection by high performance liquid chromatography, the peak area of ​​andrographolide of the standard was used as a reference and multiplied by the correction factor, as shown in Table 4. The contents of andrographolide, neoandrographolide, 14-deoxyandrographolide and dehydroandrographolide in the sample to be tested were calculated according to the pharmacopoeia.

[0069] Table 4 HPLC detection of andrographolide compounds relative retention time and correction factor

[0070]

[0071] The content of andrographolide, neoandrographolide, 14-deoxyandrographolide and dehydroandrographolide was determined by HYPERBERE C18 chromatographic column (250 mm × 4.6 mm, 5 μm, Shimadzu Corporation, Beijing) with dual wavelength detector and workstation software. About 5 μL of the sample with the specified concentration was injected into the high performance liquid chromatography column, and the gradient elution was acetonitrile (A)-pure water (B): 0-15 min, 20-25% A; 15-30 min, 25-28% A; 30-60 min, 28-40% A; 60-65 min, 40-85% A; detection wavelength: 205 nm, flow rate: 1 mg / mL.

[0072] The test results are shown in Table 5. The four fungal elicitor components of the genus AP12 can promote the accumulation of effective ingredients in Andrographis paniculata plants. Among them, the IMS component has the highest and most significant content of andrographolide, neoandrographolide, 14-deoxyandrographolide and total lactones, and the total lactone content has reached 57.27±5.47mg / g DW; the total lactone content of the MS component has reached 52.09±4.24mg / g DW, which is 1.44 times that of the CK group; the dehydroandrographolide content of the IFS component is the highest, reaching 0.41±0.23mg / gDW, which is 1.86 times that of the CK group. This shows that the genus AP12 has a significant promoting effect on the accumulation of andrographolide content.

[0073] Table 5 Effects of endophytic fungus AP12 on the content of active ingredients in Andrographis paniculata plants after 65 days of inoculation (n=3)

[0074]

[0075]

[0076] Note: CK: blank group, IFS: sterilized fermentation liquid; FS: fermentation liquid; IMS: sterilized mycelial liquid; MS: mycelial liquid. Different lowercase letters in the same column represent significant differences (p < 0.05) (statistically significant).

[0077] In summary, the endophytic fungus AP12 provided by the present invention is obtained by separation and purification from the leaf parts of Andrographis paniculata. Studies have shown that the endophytic fungus AP12 can increase the leaf area of ​​Andrographis paniculata, improve the fresh weight and dry weight, increase the accumulation of dry matter content, and significantly promote the growth of Andrographis paniculata plants; and can further promote the accumulation of the contents of four andrographolide substances. The endophytic fungus AP12 is a beneficial microorganism that can promote the growth and metabolism of Andrographis paniculata.

[0078] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. An endophytic fungus strain of Colletotrichum sp. AP12 from Andrographis paniculata, characterized in that: This strain was deposited in Guangdong Provincial Microbiological Culture Collection Center on June 15, 2023, with the deposit number GDMCC No: 63546.

2. Use of the Colletotrichum AP12 strain or its bacterial solution according to claim 1 in promoting the growth of Andrographis paniculata.

3. Use of the Colletotrichum AP12 strain or its bacterial solution according to claim 1 in increasing the content of active ingredients in Andrographis paniculata.

4. Use of the Colletotrichum AP12 strain or its bacterial solution according to claim 1 in preparing an Andrographis paniculata growth-promoting agent or in preparing a preparation for increasing the content of Andrographis paniculata active ingredients.

5. A growth-promoting agent of Andrographis paniculata and / or a preparation for increasing the content of Andrographis paniculata active ingredients, characterized in that: Containing the Colletotrichum AP12 strain or its bacterial liquid as described in claim 1.

6. A method for increasing the content of active ingredients in Andrographis paniculata, characterized in that: The Andrographis paniculata AP12 strain or its bacterial liquid as claimed in claim 1 is used to treat Andrographis paniculata.

7. The method according to claim 6, characterized in that: The bacterial liquid is fermentation liquid, inactivated fermentation liquid, mycelium liquid or inactivated mycelium liquid.

8. A method for promoting the growth of Andrographis paniculata, characterized in that: The Andrographis paniculata AP12 strain or its bacterial liquid as claimed in claim 1 is used to treat Andrographis paniculata.

9. The use according to claim 3 or 4, characterized in that: The effective ingredient of Andrographis paniculata is total andrographolide.

10. The method according to claim 6 or 7, characterized in that: The effective ingredients of Andrographolide are andrographolide, neoandrographolide, 14-deoxyandrographolide and / or dehydroandrographolide.

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