Bacillus aryabhattai FS109 and application thereof in cultivation of eucommia ulmoides and fructus forsythiae

Abstract

The invention discloses a strain of bacillus aryabhattai FS109, which is named as bacillus aryabhattai FS109, is preserved in the China Center for Type Culture Collection (CCTCC), and has the preservation number of CCTCC No: M 20242871. The strain can promote the photosynthesis of eucommia ulmoides and fructus forsythiae, promote the growth of root systems and overground parts and improve the content of active ingredients of medicine parts, and has an important effect on regulating the growth of medicinal plants and the quality of medicinal materials.

Description

Technical Field

[0001] The invention relates to Bacillus arnoldii FS109 and application thereof in the cultivation of eucommia bark and forsythia suspensa. Background Art

[0002] Eucommia ulmoides is a plant of the Eucommia family. Its dried bark and leaves are used as medicine. It has the effects of nourishing the liver and kidneys, strengthening the bones and tendons, and calming the fetus. The Shennong's Herbal Classic lists Eucommia as a top-grade product. In 2018, it was included in the "List of Chinese Medicines that Can Be Used as Health Foods" by the National Health Commission. In modern pharmacological studies, Eucommia has a positive effect on preventing hypertension, hyperglycemia, obesity, osteoporosis, etc. The oil content of Eucommia seeds is about 7-15%. Linolenic acid, oleic acid and linoleic acid are its main types of unsaturated fatty acids, accounting for 56-63%, 15-18% and 12-15% of the total fatty acid content, respectively.

[0003] Forsythia suspensa is a deciduous shrub of the Oleaceae family. Its fruit is the traditional Chinese medicine Forsythia suspensa. Forsythia suspensa is one of the 40 major medicinal materials commonly used in my country. Forsythia suspensa has the functions of clearing away heat and detoxifying, reducing swelling and dispersing stagnation, and evacuating wind-heat.

[0004] Eucommia ulmoides and Forsythia suspensa are important medicinal plants. Modern Chinese medicinal material cultivation is accompanied by excessive use of pesticides and fertilizers, resulting in quality decline, pesticide residues, heavy metal pollution, continuous cropping obstacles and other prominent problems. The ecological cultivation of Chinese medicinal materials can solve the excessive use of fertilizers to a certain extent, thereby improving the quality of medicinal materials. Rhizosphere growth-promoting bacteria can be used for the ecological cultivation of Chinese medicinal materials, cultivate high-quality Chinese medicinal materials, and directly or indirectly affect plant growth, nutrient absorption, biological or abiotic stress, etc. Possible growth-promoting mechanisms include secreting plant growth hormones, co-fixing nitrogen with plants, dissolving phosphates in the soil, improving resistance to play a biological control role, and promoting plant growth. Therefore, the development of good-performing Eucommia ulmoides and / or Forsythia suspensa rhizosphere growth-promoting bacteria will greatly promote the good growth of these medicinal plants. Summary of the invention

[0005] The present invention provides a strain of Bacillus aryabhattai FS109, named as Bacillus aryabhattai FS109, which is deposited in China Center for Type Culture Collection, the address of the depository unit is Wuhan University, Wuhan, China, the deposit time is December 20, 2024, and the deposit number is: CCTCC No: M 20242871.

[0006] The Bacillus arguta FS109 of the present invention is collected from the Lingbao Forsythia planting base in Henan Province. After the FS109 strain is cultured on a NA plate for 24 hours, the colonies formed are light yellow-white in color, smooth, dull, round, and non-fluid. The Gram staining is purple, and the colony diameter is 4-5 mm.

[0007] The physiological and biochemical characteristics of FS109 strain were as follows: VP assay was positive, MR assay was negative, citrate test was negative, propionate test was negative, nitrate reduction was positive, starch hydrolysis was positive, D-xylose test was positive, D-mannitol test was positive, anaerobic growth was positive, 7% NaCl growth was negative, and pH5.7 growth was negative.

[0008] The tested 16SrDNA sequence was compared with the sequences in the GenBank database. The results showed that FS109 and Bacillus aryabhattai were on the same branch, and its 16S rDNA sequence was 100% similar to Bacillus aryabhattai (MT538470). Combined with the colony morphology, physiological and biochemical characteristics and 16SrDNA sequence analysis, it was identified as Bacillus aryabhattai.

[0009] The experiment found that the strain was in a slow growth phase from 0 to 6 hours, in a logarithmic growth phase from 6 to 18 hours, and reached a peak value when the bacterial volume reached 30 hours, so the best culture time was 30 hours. The best culture medium was 1.7g lactose, 2.4g beef extract powder, 1.5g MgSO4, 20g agar, 1000ml water, pH 7, and the best inoculation amount was 2.0% of the culture medium. The best culture conditions were a temperature of 35°C and a rotation speed of 180r·min -1 .

[0010] The beneficial effects achieved by the present invention are as follows: the Bacillus FS109 isolated from the rhizosphere of Forsythia suspensa in the present invention can promote photosynthesis, promote the growth of the root system and the above-ground parts, and increase the content of active ingredients in the medicinal parts when acting on Eucommia ulmoides and Forsythia suspensa, which plays an important role in regulating the growth of medicinal plants and the quality of medicinal materials. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention and do not constitute a limitation of the present invention. In the accompanying drawings:

[0012] Figure 1 The effect of FS109 treatment on the chlorophyll content in Eucommia ulmoides leaves;

[0013] Figure 2 The changes of chlorophyll content in potted young Forsythia suspensa seedlings after FS109 treatment;

[0014] Figure 3 The changes of chlorophyll content in Forsythia suspensa plants after FS109 treatment;

[0015] Different letters in the figure indicate significant differences at the P<0.05 level by T test. DETAILED DESCRIPTION

[0016] The preferred embodiments of the present invention are described below in conjunction with the accompanying drawings. It should be understood that the preferred embodiments described herein are only used to illustrate and explain the present invention, and are not used to limit the present invention.

[0017] Example 1

[0018] 1. Isolation and identification of experimental strains

[0019] 1.1 Optimization of strain isolation and culture conditions

[0020] At the Forsythia suspensa planting base in Lingbao, Henan, five healthy Forsythia suspensa plants were selected using the five-point sampling method. The rhizosphere soil 20 cm above the ground surface was collected at a distance of about one meter from the roots of the plants as experimental materials. After taking it out, it was preserved in a plastic bag and immediately sent back to the laboratory for experiments. The collected soil samples were naturally air-dried and ground, then passed through a 20-mesh sieve and diluted 10 times with distilled water, and placed on a shaker at 180 rpm for 1 hour. After shaking and mixing, 1 mL of the soil suspension was mixed with 9 mL of sterile water and shaken to make a 10-fold diluted soil suspension. After the soil particles were allowed to settle, 1 mL of the supernatant was taken and mixed with 9 mL of sterile water to make 10 2 , 10 3 , 10 4 , 10 5 , 10 6 There are 6 suspensions with different dilution ratios. Use a pipette to dilute 10 4 , 10 5 and 10 6 200 μL of the suspension was spread on NA, LB and TSB plates respectively, and each concentration was repeated 3 times in each culture medium. After culturing at 30℃ for 2 days, colonies with different appearances were transferred. After streaking purification on NA, LB and TSB plates, all isolated strains were numbered and stored at 4℃.

[0021] 1.2 Antimicrobial activity screening

[0022] The plate confrontation method was used to screen the bacteria isolated in the laboratory. The pathogenic fungi used were Botryosphaeria dothidea and Phomopsis velata.

[0023] The pathogenic fungi and the isolated bacteria were activated separately, and the back of the PDA plate was marked with a "cross". Bacteria were inoculated around the "cross" at 2.5 cm from the center. After 2 days, pathogenic fungi with a diameter of 5 mm were inoculated at the center of the "cross" on the PDA plate. The culture was carried out in the dark at 26°C for 72 hours, and each treatment was repeated 3 times. The inhibition of bacteria on pathogenic fungi was observed, and the width of the inhibition zone between the edge of the pathogenic fungi and the edge of each bacteria was measured. Finally, the strain numbered FS109 was screened out, which had a larger width of the inhibition zone against both fungi and was a strain with antibacterial activity.

[0024] The colonies formed after culturing FS109 strain on NA plate for 24 hours are light yellow-white in color, smooth, dull, round, non-fluid, purple in Gram staining, and 4-5 mm in diameter.

[0025] Physiological and biochemical characteristics

[0026] The FS109 strain was positive in VP assay, negative in MR assay, negative in citrate test, negative in propionate test, positive in nitrate reduction, positive in starch hydrolysis, positive in D-xylose test, positive in D-mannitol test, positive in anaerobic growth, negative in 7% NaCl growth, and negative in pH5.7 growth.

[0027] The strains were identified by referring to the methods in the monographs such as Bergey's Manual of Bacterial Identification, 8th Edition and Manual of Identification of Common Bacterial Systems. PCR amplification was performed using universal primers for bacterial 16S rDNA. The upstream primer (27F) was 5'-AGAGTTTGATCCTGGCTCAG-3'; the downstream primer (1492R) was 5'-TACGGYTACCTTGTTACGACTT-3'. The PCR products were sequenced and the sequencing results were compared by BLAST to identify their species.

[0028] The tested 16SrDNA sequence was compared with the sequences in the GenBank database. The results showed that FS109 and Bacillus aryabhattai were on the same branch, and its 16S rDNA sequence was 100% similar to Bacillus aryabhattai (MT538470). Combined with the colony morphology, physiological and biochemical characteristics and 16SrDNA sequence analysis, it was identified as Bacillus aryabhattai.

[0029] 1.4 Optimization of strain culture conditions

[0030] A single colony of FS109 strain was inoculated into 10 mL of LB medium and heated at 180 r·min. -1, shake culture at 28℃ for 24h to prepare seed solution. Inoculate 1% of the seed solution into 10mL of basal culture medium and culture under the same conditions. Use sterile basal culture medium as control and measure the OD of the bacterial solution every 6h. 600 Values, 3 replicates per treatment. With time as the horizontal axis, OD 600 The value is the vertical axis, and the growth curve of BA109 strain is drawn.

[0031] Using the basal medium as the initial culture medium, the effects of the types and concentrations of carbon sources (glucose, sucrose, lactose, xylitol and mannitol), nitrogen sources (peptone, urea, beef extract powder, ammonium sulfate and ammonium chloride) and inorganic salts (sodium chloride, potassium dihydrogen phosphate, potassium chloride, magnesium sulfate and copper sulfate pentahydrate) on the growth of BA109 strain were determined. Subsequently, the effects of culture temperature, initial pH value and shaking speed on the culture of the strain were screened.

[0032] The experiment found that the strain was in a slow growth phase from 0 to 6 hours, in a logarithmic growth phase from 6 to 18 hours, and reached a peak value when the bacterial volume was cultivated for 30 hours, so the best cultivation time was 30 hours. The best culture was basically lactose 1.7g, beef extract powder 2.4g, MgSO4 1.5g, agar 20g, water 1000ml, pH 7, and the best inoculation amount was 2.0% of the culture medium. The best culture conditions were a temperature of 35°C and a rotation speed of 180r·min -1 .

[0033] 2. Eucommia ulmoides cultivation experiment

[0034] 2.1 Fertilization

[0035] Bacterial liquid culture: The isolated strain Bacillus aryabhattai FS109 was activated on the NA slant, and a loop was selected and inoculated into a special culture medium (lactose 1.7g, beef extract powder 2.4g, MgSO4 1.5g, agar 20g, water 1000ml, pH 7.3±0.1) and incubated at 35℃, 180r·min -1 The culture was shaken for 30 h under the conditions, and the culture medium was diluted with sterile water to contain 1×10 8 cfu.mL -1 The bacterial suspension was used for inoculation.

[0036] The experiment was conducted at the Mengzhou base of the Chinese Academy of Forestry. The test plants were selected from the six-year-old nationally approved variety 'Huazhong No. 8' Eucommia ulmoides plants. On April 10, 1 kg of organic fertilizer was prepared. Clean water and 100 ml of bacterial solution were mixed with organic fertilizer. Two trenches were opened under the root crown with a depth of 20 cm. The prepared organic bacterial fertilizer was applied. Five plants were treated each time, and each treatment was repeated 3 times.

[0037] The fruits were collected in October when the fruits matured, and the weight of 100 fruits was measured. The contents of forsythiaside, forsythiaside A, and volatile oil were also measured; the plant height, root length, and root width were also measured.

[0038] 2.2 Determination of chlorophyll content

[0039] After 30 days, collect 10 leaves from the 4th and 5th nodes from the middle branches of the plants from top to bottom, mix them thoroughly as a mixed sample, put them in a foam box with ice packs, and bring them back to the laboratory for use.

[0040] Chlorophyll a and chlorophyll b content determination method: Chlorophyll extraction adopts anhydrous ethanol extraction method, avoiding the leaf edges and veins, and cutting the leaves into 2mm wide threads with scissors. Soak in the dark for 12 hours until it turns white, and use a purple spectrophotometer to measure chlorophyll a and chlorophyll b at 645nm and 663nm respectively. The chlorophyll content is calculated using the Arnon method.

[0041] from Figure 1 It can be seen that after 30 days of root irrigation with Bacillus FS109, the contents of chlorophyll a and chlorophyll b in Eucommia ulmoides leaves increased significantly. Chlorophyll plays an important role in the absorption, transmission and conversion of light energy. The increase in chlorophyll content can increase the photosynthesis rate of plants, thereby promoting the synthesis of plant organic matter and promoting plant growth.

[0042] 2.3 Determination of Eucommia ulmoides plant growth indicators

[0043] Three seedlings were randomly selected from each group to measure plant biomass indicators, and the trunk and branch diameters of each group of Eucommia ulmoides after 0 and 150 days of cultivation were recorded respectively. Each treatment was repeated three times.

[0044] Diameter determination of trunk and branches: Three plants were randomly selected from each group, and the diameter of the branches at 1 cm from the trunk was measured using a vernier caliper. The diameter data was in mm, with two decimal places. Three plants were selected from each group.

[0045] Table 1 Effects of FS109 treatment on the growth of Eucommia ulmoides trunk and current year branches

[0046]

[0047] As can be seen from Table 1, the diameter growth of the main trunk of Eucommia ulmoides treated with Bacillus fertigation FS109 was significantly higher than that of the untreated main trunk of Eucommia ulmoides.

[0048] 2.4 Determination of active ingredients in leaves

[0049] Determination of the content of aucubin and chlorogenic acid: Weigh a certain amount of Eucommia ulmoides peel sample, crush and sieve it, accurately weigh about 2.0g and place it in a stoppered conical flask, add an appropriate amount of methanol (solid-liquid ratio is 1:20), ultrasonically extract in an ultrasonic cleaner for 30 minutes, centrifuge the extract and filter the supernatant to obtain the sample solution to be tested. Then use high performance liquid chromatography to determine, set the chromatographic column to a C18 reverse phase column, the mobile phase is acetonitrile-0.01% phosphoric acid water (volume ratio is 6:94), the flow rate is 0.8mL / min, the column temperature is 30°C, and the detection wavelength is 254nm; first draw a standard curve, weigh the aucubin standard and dissolve it in methanol to prepare standard solutions of different concentrations, inject it into a high performance liquid chromatograph to record the peak area, draw a standard curve with concentration as the horizontal axis and peak area as the vertical axis, and then take an appropriate amount of the sample solution to be tested and inject it into the high performance liquid chromatograph to record the peak area and calculate the content and chlorogenic acid content in the sample according to the standard curve. The experimental results are shown in Table 3.

[0050] 2.5 Determination of Eucommia ulmoides fruit growth index, Eucommia ulmoides oil and Eucommia ulmoides gum content

[0051] Determination of fruit growth index: Reference standard GB / T 3543.7-1995 Crop Seed Inspection Procedure. On October 20, when the fruits were ripe, 3 Eucommia fruit samples were randomly picked from each group and placed in numbered ziplock bags. Some were stored in a 4℃ refrigerator to measure the morphological characteristics of Eucommia fruit, including 100-grain weight, water content, transverse diameter, etc. Some were directly dried at 55℃ for later use.

[0052] After the samples were fully mixed, they were randomly sampled using the quartering method, with 100 fruits in each portion, and weighed using an electronic balance (accuracy: 0.01g), which was the 100-grain weight. Ten fruits were randomly selected from each seedling using the quartering method, and the vertical and horizontal diameters of the fruits were measured using a vernier caliper (accuracy: 0.01mm).

[0053] Determination of Eucommia seed oil content: Weigh a certain amount of Eucommia seed sample, crush and sieve it, weigh 5.000 grams of fully mixed Eucommia seed kernels and put them into a filter paper tube, then place the filter paper tube in the extraction tube of the Soxhlet extractor, and connect the receiving bottle with constant weight. Add 100 ml of petroleum ether from the upper end of the condenser tube, heat and reflux in an oil bath for 2 hours, use a frosted glass rod to receive 1 drop of extract, and the extraction is complete when there is no oil spot. Remove the receiving bottle, use a rotary evaporator (80°C, 60rpm, vacuum degree 0.07-0.08MPa, about 10 minutes) to reduce pressure and recover the solvent, cool and weigh to calculate the yield, repeat the same conditions 3 times and take the average value.

[0054] Determination of eucommia gum content: accurately weigh 5g of eucommia peel powder, add 100ml petroleum ether solvent to the Soxhlet extractor, heat and reflux in a water bath at 80-85℃ for 2h, then use a 120-mesh stainless steel mesh as a filter, filter the reflux mixture while hot, separate the residue, and collect the filtrate dissolved with eucommia gum in a stoppered triangular flask. After the filtrate dissolved with eucommia gum is cooled by water, it is placed in a low-temperature (-20℃) storage box for about 1h, and filtered with a 120-mesh stainless steel mesh to obtain eucommia gum and the filtrate without eucommia gum. The filtrate is returned and mixed with the residue, and the residual eucommia gum is completely dissolved by constant temperature reflux for 1-2h again. The conditions and operation are the same as the first time. The recovered eucommia gum is combined with the eucommia gum precipitated for the first time to obtain the product, and its total mass is accurately weighed.

[0055] As shown in Table 2, after the root irrigation with Bacillus fertigation FS109, the fruit of Eucommia ulmoides was significantly enlarged, as shown by the longitudinal diameter of the treated fruit being 35.98 mm, an increase of 17.3% compared with the untreated fruit, and the weight of 100 dry fruits being 8.92 g, an increase of 23.2% compared with the control fruit of 7.24 g.

[0056] Table 2 Effect of FS109 treatment on the growth of Eucommia ulmoides fruit

[0057]

[0058]

[0059] The contents of chlorogenic acid and aucubin in leaves, and the contents of volatile oil and eucommia gum in fruits were determined. The results shown in Table 3 showed that after root irrigation with Bacillus arguta FS109, the chlorogenic acid and aucubin in leaves increased by 30.3% and 23.0%, respectively, with significant differences. However, the contents of volatile oil and eucommia gum in fruits did not change significantly.

[0060] Table 3 Effects of FS109 on important components of Eucommia ulmoides leaves and fruits

[0061]

[0062] 3 Forsythia suspensa cultivation experiment

[0063] 3.1 Fertilization

[0064] Potted seedlings: This experiment was conducted at the School of Pharmacy of Henan University. At the end of October, the Forsythia suspensa seedlings that had fallen leaves and were grafted in August of that year were selected and transplanted into nutrient pots. 8 cfu.mL -1 The roots of the forsythia seedlings were inoculated with the bacterial suspension of 20 mL / plant, with 10 plants treated for 3 times, and clean water was used as the control. The forsythia seedlings were placed in a constant temperature greenhouse with a light intensity of 2500 lux and a daylight length of 12 hours / day.

[0065] Field plants: The experiment was conducted at the Forsythia suspensa planting base of Sanmenxia Damei Co., Ltd. in Lingbao City. In April 2024, three-year-old Forsythia suspensa plants were selected, the variety was Lingqiao No. 1, two trenches were opened under the root crown of the plant, the trench depth was 15 cm, and 50 mL of bacterial suspension was poured in. Ten plants were treated each time, and each treatment was repeated 3 times, with clean water as the control.

[0066] 3.2 Chlorophyll content determination

[0067] Potted seedlings: At 50 days old, collect 10 leaves from each plant and mix them thoroughly as a mixed sample for later use.

[0068] Field plants: On August 15, collect 10 leaves from the 4th and 5th nodes from the middle branches of the plants from top to bottom, mix them thoroughly as a mixed sample, put them in a foam box with ice packs, and bring them back to the laboratory for use.

[0069] Chlorophyll a and chlorophyll b content determination method: Chlorophyll extraction adopts anhydrous ethanol extraction method, avoiding the leaf edges and veins, and cutting the leaves into 2mm wide threads with scissors. Soak in the dark for 12 hours until it turns white, and use a purple spectrophotometer to measure chlorophyll a and chlorophyll b at 645nm and 663nm respectively. The chlorophyll content is calculated using the Arnon method.

[0070] from Figure 2 and Figure 3 It can be seen that after 30 days of root irrigation with Bacillus FS109, the contents of chlorophyll a and chlorophyll b in the leaves of Forsythia suspensa seedlings and field plants increased significantly. Chlorophyll plays an important role in the absorption, transmission and conversion of light energy. The increase in chlorophyll content can increase the photosynthesis rate of plants, thereby promoting the synthesis of plant organic matter and promoting plant growth.

[0071] 3.3 Determination of plant growth indicators

[0072] Potted seedlings: At 50 days, plant height, root length, root width and fresh weight of aboveground parts were measured.

[0073] Field plants: On August 15, when the fruits were just beginning to ripen, the branch growth was measured.

[0074] As can be seen from Table 4, compared with the water control group, after 30 days of root irrigation with Bacillus FS109, the growth of Forsythia suspensa seedlings was significantly accelerated, and the number of nodes, plant height, root length, root width and aboveground fresh weight were significantly higher than those of the control. After the application of bacteria, the number of nodes was 8.2, an increase of 29.0%; the plant height was 10.6cm, an increase of 20.5%; the root length was 15.37, an increase of 14.7%; the aboveground fresh weight was 3.36, an increase of 39.4%.

[0075] Table 4 Effect of FS109 on the growth of Forsythia suspensa potted seedlings

[0076]

[0077] As can be seen from Table 5, compared with the water control group, after 30 days of root irrigation with Bacillus FS109, the leaf length and width of Forsythia suspensa plants in the field did not increase significantly. However, the trunk growth was significantly different from the water control group. More importantly, the fruit length increased by 10.3% and the fruit 100 seeds increased by 11.4% compared with the water control group.

[0078] 3.4 Determination of fruit growth indicators

[0079] On August 15, when the fruits were just beginning to ripen, the fruits were collected and brought back to the laboratory for later use. The fruit size and 100-grain weight of Forsythia suspensa were measured.

[0080] Table 5 Effects of FS109 treatment on plant growth and fruit size of Forsythia suspensa in the field

[0081]

[0082] 3.5 Determination of active ingredient content

[0083] Potted seedlings: Take the leaves of Forsythia suspensa seedlings and determine the content of forsythiaside and forsythiaside A: accurately weigh 2 mg of forsythiaside or forsythiaside A reference substance, add 2 mL of methanol, and make a solution with a concentration of 1.0 mg per 1 ml. Take about 2 g of fruit powder (passed through a No. 5 sieve), accurately weigh it, put it in a stoppered conical bottle, accurately add 25 ml of methanol, weigh it, ultrasonically treat it (power 250 W, frequency 40 kHz) for 25 minutes, let it cool, weigh it again, make up the lost weight with methanol, shake it well, filter it, and take the filtrate to obtain it. Determine the content of forsythiaside. Take about 0.5g of the powder of this product (passed through a No. 5 sieve), weigh it accurately, put it in a stoppered conical bottle, add 15ml of 70% methanol accurately, stopper it tightly, weigh it, ultrasonically treat it (power 250W, frequency 40kHz) for 30 minutes, let it cool, weigh it again, make up the lost weight with 70% methanol, shake it well, filter it, and take the filtrate to obtain it. Determine the content of forsythiaside A. The experimental data were tested for significant differences using the SPSS T test method.

[0084] Field: Take Forsythia suspensa fruits and determine the contents of forsythiaside and forsythiaside A.

[0085] Experimental data analysis

[0086] The significance of the difference was tested using the SPSS T test.

[0087] It can be seen from Table 6 that the intrinsic quality of Forsythia suspensa fruit has been significantly improved compared with the clear water control group. After root irrigation with Bacillus argei FS109, the content of forsythia glycoside reached 0.29%, while that of the clear water control was 0.22%, an increase of 31.8%; the content of forsythia glycoside A reached 10.7%, while that of the clear water control was 8.1%, an increase of 32.1. There was no significant change in the content of volatile oil. The failure of forsythia glycosides in old Forsythia suspensa to meet the standards is a problem that plagues Chinese medicine processing companies. The 2020 edition of the "Chinese Pharmacopoeia" stipulates that the content of forsythia glycosides shall not be less than 0.15%. As time goes on, the forsythia glycosides in Forsythia suspensa will degrade. The Bacillus argei FS109 of this patent can increase the forsythia glycosides in green Forsythia suspensa by 31.8%, which can provide a guarantee for the later production of qualified old Forsythia suspensa.

[0088] Table 6 Effects of FS109 treatment on active ingredients in fruits of Forsythia suspensa field plants

[0089]

[0090] In summary, the antibiotic strain Bacillus aryabhattai FS109 was used in Eucommia ulmoides and Forsythia suspensa. It can be seen that this strain can promote plant photosynthesis, promote the growth of roots and aboveground parts, increase the content of active ingredients in medicinal parts, and play an important role in regulating the growth of medicinal plants and the quality of medicinal materials.

[0091] Finally, it should be noted that the above is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. Bacillus aryabhattai FS109, named Bacillus aryabhattai FS109, is deposited in China Center for Type Culture Collection, the address of the depository is Wuhan University, Wuhan, China. The deposit date is December 20, 2024, and the deposit number is: CCTCC No: M 20242871.

2. The Bacillus arifugei FS109 according to claim 1, characterized in that The culture medium is lactose 1.7g, beef extract powder 2.4g, MgSO4 1.5g, agar 20g, water 1000ml, pH7.

3. The Bacillus arifugei FS109 according to claim 1 or 2, characterized in that The culture conditions were 35°C and 180 rpm. -1 .

4. Use of Bacillus arifugeyi FS109 in promoting plant photosynthesis.

5. Use of Bacillus arifugeyi FS109 in promoting plant growth.

6. The use according to claim 4 or 5, characterized in that The plant is Eucommia ulmoides or Forsythia suspensa.

7. Use of Bacillus arifuge FS109 in promoting the increase of active ingredients in Eucommia ulmoides or Forsythia suspensa.

8. The use according to claim 7, characterized in that The active ingredient is any one or more of chlorogenic acid, aucubin, forsythiaside or forsythiaside A.

9. The use according to claim 7 or 8, characterized in that Pour the bacterial solution of Bacillus arifugeti FS109 into the root crown of Eucommia ulmoides or Forsythia suspensa.

10. The use according to claim 9, characterized in that The concentration of Bacillus FS109 was 1×10 8 cfu.mL -1 , the dosage is 20-100mL.

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