Bacillus aryabhattai fs109 and its application in eucommia ulmoides and forsythia suspensa cultivation

Abstract

The application discloses a bacillus aryabhattai FS109, named as Bacillus aryabhattai FS109, which is preserved in the China Center for Type Culture Collection, and the preservation number is CCTCC No: M 20242871. The strain can promote photosynthesis of eucommia ulmoides and forsythia suspensa, promote growth of root systems and aboveground parts, and increase content of active ingredients in medicinal parts, and has an important role in regulating growth of medicinal plants and quality of medicinal materials.

Description

TECHNICAL FIELD

[0001] The application relates to Bacillus aryabhattai FS109 and application thereof in Eucommia ulmoides and Forsythia suspensa cultivation. BACKGROUND

[0002] Eucommia ulmoides is a plant in Eucommiaceae, and the dried bark and leaves are used as medicines, and have the effects of tonifying liver and kidney, strengthening muscles and bones, and preventing miscarriage. Eucommia ulmoides is listed as a superior product in Shennong's Herbal Classic. In 2018, it was included in the list of Chinese medicines that can be used for health foods by the National Health Commission of China. In modern pharmacological research, Eucommia ulmoides has a positive effect on preventing hypertension, hyperglycemia, obesity, osteoporosis and the like. The oil content of Eucommia ulmoides seed is about 7-15%. Linolenic acid, oleic acid and linoleic acid are main unsaturated fatty acid types, accounting for 56-63%, 15-18% and 12-15% of the total fatty acid content, respectively.

[0003] Forsythia suspensa is a deciduous shrub in Oleaceae, and the fruit is a traditional Chinese medicine Forsythia. Forsythia is one of the 40 most commonly used medicinal materials in China. Forsythia has the effects of clearing heat and resolving toxins, reducing swelling and resolving nodes, and dispersing wind-heat.

[0004] Eucommia ulmoides and Forsythia are important medicinal plants. With the excessive use of pesticides and fertilizers in modern Chinese medicinal material cultivation, the quality of medicinal materials is reduced, pesticide residues, heavy metal pollution and continuous cropping obstacles are prominent. Ecological cultivation of Chinese medicinal materials can solve the problem of excessive use of fertilizers to some extent and improve the quality of medicinal materials. Rhizosphere growth-promoting bacteria can be used for ecological cultivation of Chinese medicinal materials to cultivate high-quality Chinese medicinal materials, directly or indirectly affecting plant growth, nutrient absorption, biological or non-biological stress and the like. Possible growth-promoting mechanisms include secretion of plant growth hormones, joint nitrogen fixation with plants, dissolution of phosphate in soil, improvement of resistance for biocontrol and promotion of plant growth. Therefore, development of Eucommia ulmoides and / or Forsythia rhizosphere growth-promoting bacteria with good performance will greatly promote the growth of these medicinal plants. SUMMARY

[0005] The application provides a Bacillus aryabhattai FS109, named as Bacillus aryabhattai FS109, which is preserved in the China Center for Type Culture Collection, has a preservation unit address of China.Wuhan.Wuhan University, a preservation time of December 20, 2024 and a preservation number of CCTCC No: M 20242871.

[0006] The Bacillus aryabhattai FS109 of the application is collected from a Forsythia suspensa planting base in Lingbao, Henan. The colony formed after the FS109 strain is cultured on a NA plate for 24 hours is light yellow white in color, smooth in surface, non-lustrous, round in shape, non-flowing, purple in gram staining and 4-5 mm in colony diameter.

[0007] The physiological and biochemical characteristics are as follows: V.P assay of FS109 strain is positive, MR assay is negative, citrate test is negative, propionate test is negative, nitrate reduction is positive, starch hydrolysis is positive, D-xylose test is positive, D-mannitol test is positive, anaerobic growth is positive, 7% NaCl growth is negative, and pH 5.7 growth is negative.

[0008] The test 16S rDNA sequence is compared with sequences in the GenBank database, and the result shows that FS109 is in the same branch as Bacillus aryabhattai, and the similarity of the 16S rDNA sequence with Bacillus aryabhattai (MT538470) is 100%. In combination with colony morphology, physiological and biochemical characteristics and 16S rDNA sequence analysis, it is identified as Bacillus aryabhattai.

[0009] It is found through experiments that the strain is in a slow growth period at 0-6h, is in a logarithmic growth period at 6-18h, and the cell amount reaches a peak at 30h of culture, so that the optimal culture time is 30h. The optimal culture medium is 1.7g of lactose, 2.4g of beef extract powder, 1.5g of MgSO4, 20g of agar and 1000ml of water, pH 7, and the optimal inoculation amount is 2.0% of the culture medium. The optimal culture condition is 35 DEG C of temperature and 180r / min of rotation speed. -1 .

[0010] The application has the beneficial effects that the Bacillus aryabhattai FS109 separated from the rhizosphere of Forsythia suspensa and applied to Eucommia ulmoides and Forsythia suspensa can promote photosynthesis, promote the growth of root systems and aboveground parts, and improve the content of active ingredients in medicinal parts, and has an important role in regulating the growth of medicinal plants and the quality of medicinal materials. BRIEF DESCRIPTION OF DRAWINGS

[0011] The accompanying drawings are included to provide a further understanding of the application, and constitute a part of the specification, illustrate the application, and are used to explain the application together with the embodiments of the application, and do not constitute a limitation on the application. In the drawings:

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

[0013] Figure 2 Chlorophyll content change of potted young Forsythia suspensa seedlings after FS109 treatment;

[0014] Figure 3 Chlorophyll content change of field Forsythia suspensa plants after FS109 treatment;

[0015] Wherein, different letters in the figure represent significant difference at P < 0.05 level by T test. DETAILED DESCRIPTION

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

[0017] Example 1

[0018] 1. Isolation and identification of experimental strains

[0019] 1.1 Optimization of strain isolation and culture conditions

[0020] In Henan Lingbao forsythia planting base, five healthy forsythia were selected by five-point sampling method, and rhizosphere soil 20 cm away from the ground was taken about 1 meter away from the plant roots as experimental material. After taking out, it was stored in a plastic bag, and immediately sent back to the laboratory for experiment. The collected soil sample was naturally dried, ground, passed through a 20 mesh sieve, and diluted 10 times with distilled water, and placed in a 180 rpm shaker for 1 h. After mixing, 1 mL of soil suspension was mixed with 9 mL of sterile water to prepare a 10-fold diluted soil suspension. After the soil particles were allowed to settle, 1 mL of supernatant was taken and mixed with 9 mL of sterile water, and 10 2 , 10 3 , 10 4 , 10 5 , 10 6 Suspensions of 6 different dilutions were prepared. 200 μL of 10 4 , 10 5 and 10 6 fold dilutions of the suspension were used to coat NA, LB and TSB plates, respectively, with 3 repeats for each concentration in each medium. After incubation at 30°C for 2 days, different colonies were transferred. After NA, LB and TSB plate streaking, all isolated strains were numbered and stored at 4°C.

[0021] 1.2 Screening of antibacterial activity

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

[0023] The pathogenic fungi and the isolated bacteria were activated respectively, marked with a "cross" on the back of the PDA plate, inoculated with bacteria at a distance of 2.5 cm from the center around the "cross", and after 2 days, inoculated with pathogenic fungi with a diameter of 5 mm at the center point of the "cross" of the PDA plate, and cultured at 26°C in the dark for 72 hours, with 3 repeated treatments each time. The inhibition of the bacteria on the 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 on two kinds of fungi, and was a strain with antibacterial activity.

[0024] The colony formed by the FS109 strain after being cultured on the NA plate for 24 hours was light yellow white in color, smooth in surface, non-lustrous, round in shape, non-flowing, purple in gram staining, and 4-5 mm in diameter.

[0025] Physiological and biochemical characteristics

[0026] The V.P. determination of the FS109 strain was positive, the MR determination was negative, the citrate test was negative, the propionate test was negative, the nitrate reduction was positive, the starch hydrolysis was positive, the D-xylose test was positive, the D-mannitol test was positive, the anaerobic growth was positive, the 7% NaCl growth was negative, and the pH 5.7 growth was negative.

[0027] The strain was identified by referring to the methods in the monographs such as "Berger's Bacterial Identification Manual · 8th Edition" and "Common Bacterial System Identification Manual". PCR amplification was performed by using bacterial 16S rDNA universal primers. The upstream primer (27F) was 5'-AGAGTTTGATCCTGGCTCAG-3', and the downstream primer (1492R) was 5'-TACGGYTACCTTGTTACGACTT-3'. The PCR product was sequenced, and the sequencing results were subjected to BLAST comparison to identify the species.

[0028] The test 16S rDNA sequence was compared with the sequences in the GenBank database, and the results showed that the FS109 was in the same branch as Bacillus aryabhattai, and the similarity of the 16S rDNA sequence thereof to Bacillus aryabhattai (MT538470) was 100%. In combination with the colony morphology, physiological and biochemical characteristics, and 16S rDNA sequence analysis, it was identified as Bacillus aryabhattai.

[0029] 1.4 Optimization of strain culture conditions

[0030] The FS109 strain was inoculated with a single colony into 10 mL of LB medium, and cultured at 37°C, 180 r·min-1, and 220 r·min-1 for 24 hours, and then the culture was centrifuged at 4°C and 8000 r·min-1 for 5 minutes, and the supernatant was discarded. -1, 28℃, 180r·min-1, for 24h to make seed liquid. The seed liquid was inoculated into 10mL basic culture medium with 1% inoculation amount, and cultured under the same condition, taking sterile basic culture medium as control. OD 600 value was measured every 6h, and each treatment was repeated 3 times. The growth curve of BA109 strain was drawn with time as horizontal axis and OD 600 value as vertical axis.

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

[0032] It was found through experiments that the strain was in slow growth phase at 0-6h, in logarithmic growth phase at 6-18h, and the cell amount reached peak value at 30h of culture, so the optimal culture time was 30h. The optimal culture base was lactose 1.7g, beef extract 2.4g, MgSO4 1.5g, agar 20g, water 1000ml, pH7, and the optimal inoculation amount was 2.0% of culture medium. The optimal culture condition was temperature 35℃, shaking speed 180r·min -1 .

[0033] 2, Eucommia cultivation experiment

[0034] 2.1 Fertilization

[0035] Liquid culture: the isolated strain Bacillus aryabhattai FS109 was activated on NA slant, and a ring was inoculated into special culture medium (lactose 1.7g, beef extract 2.4g, MgSO4 1.5g, agar 20g, water 1000ml, pH7.3±0.1) and cultured at 35℃, 180r·min -1 for 30h. The culture liquid was diluted with sterile water to prepare a bacterial suspension containing 1×10 8 cfu·mL -1 for inoculation.

[0036] The experiment was carried out at the Mengzhou base of Chinese Academy of Forestry, and six-year-old national standard variety 'Huazhong No.8' eucommia plants were selected as test plants. On April 10th, 1kg of organic fertilizer was prepared, and the water and 100ml of bacterial liquid were mixed with the organic fertilizer, respectively. Two trenches were dug under the root crown, with a depth of 20cm, and the prepared organic bacterial fertilizer was applied, with 5 plants per treatment and 3 repeated treatments.

[0037] The fruits were collected in the fruit ripening period, and the hundred-grain weight of the fruits was determined, and the forsythia fruit, forsythia ester A, and volatile oil contents were determined.

[0038] 2.2 Chlorophyll content determination

[0039] After 30 days, 10 leaves from the 4th to 5th nodes of the middle branches of the plants were collected from top to bottom, mixed thoroughly as a mixed sample, and placed in a foam box with an ice bag for transportation back to the laboratory for standby.

[0040] The chlorophyll a and chlorophyll b content determination method: chlorophyll extraction uses anhydrous ethanol extraction method, avoiding the edges and veins of the leaves, and cutting the leaves into 2 mm wide filaments with scissors. Dark soak for 12 h until white, and measure the chlorophyll a and chlorophyll b at 645 nm and 663 nm respectively using a purple spectrophotometer. The chlorophyll content is calculated using the Arnon method.

[0041] As can be seen from Table 1, the diameter growth of the main stem of Eucommia ulmoides treated with Bacillus aryabhattai FS109 was significantly higher than that of the untreated Eucommia ulmoides. Figure 1 It can be seen that the chlorophyll a and chlorophyll b contents in the leaves of Eucommia ulmoides were significantly increased after 30 days of Bacillus aryabhattai FS109 root irrigation treatment. Chlorophyll plays an important role in light energy absorption, transmission, and conversion, and an 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 for measuring plant biomass indicators, and the main stem branch diameters of Eucommia ulmoides in each group were recorded after 0 days and 150 days of culture, and each treatment was repeated three times.

[0044] Main stem branch diameter determination: three plants to be measured were randomly selected from each group, and the diameter at 1 cm from the main stem of the branch was measured using a vernier caliper. The diameter data is in mm, with two decimal places. Three plants were selected from each group.

[0045] Table 1 Effect of FS109 treatment on the growth of the main stem and current year branches of Eucommia ulmoides

[0046]

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

[0048] 2.4 Leaf active ingredient determination

[0049] Aucubin and chlorogenic acid content determination: a certain amount of Eucommia bark sample was crushed and sieved, and about 2.0 g was accurately weighed and placed in a conical flask with a plug. Appropriate amount of methanol (1:20 of solid-liquid ratio) was added, and the sample was ultrasonically extracted for 30 minutes. The supernatant was obtained by centrifugation and filtration to obtain the sample solution. Then, high performance liquid chromatography was used for determination. The chromatographic column was C18 reversed-phase chromatographic column, the mobile phase was acetonitrile-0.01% phosphoric acid (volume ratio was 6:94), the flow rate was 0.8 mL / min, the column temperature was 30°C, and the detection wavelength was 254 nm. First, the standard curve was drawn. The aucubin standard was dissolved in methanol to prepare standard solutions of different concentrations. The peak area was recorded by injecting the high performance liquid chromatograph. The standard curve was drawn with concentration as the abscissa and peak area as the ordinate. Then, the peak area of the sample solution was recorded by injecting the high performance liquid chromatograph, and the content of aucubin and chlorogenic acid in the sample was calculated according to the standard curve. The experimental results are shown in Table 3.

[0050] 2.5 Growth index of Eucommia fruit, Eucommia oil and Eucommia gum content determination

[0051] Fruit growth index determination: refer to the standard GB / T 3543.7-1995 Crop Seed Test Rules. On October 20th, when the fruit is mature, 3 Eucommia fruit samples are randomly taken from each group and placed in self-sealing bags. A part is placed in a 4°C refrigerator for storage, and used to measure the morphological characteristics of Eucommia fruit, such as hundred-grain weight, water content, and transverse diameter. A part is directly dried at 55°C for standby.

[0052] After the sample is thoroughly mixed, random sampling is performed by quartering method, and 100 grains are taken from each part. The weight is measured by an electronic balance (accuracy: 0.01 g), which is the hundred-grain weight. Ten fruits are randomly selected from each seedling by quartering method, and the fruit longitudinal diameter and transverse diameter are measured by a vernier caliper (accuracy: 0.01 mm).

[0053] Determination of Eucommia seed oil content: a certain amount of Eucommia seed sample was crushed and sieved, and 5.000 g of thoroughly mixed Eucommia seed kernel was weighed and placed in a filter paper cylinder. The filter paper cylinder was then placed in a Soxhlet extractor extraction cylinder, and a constant weight receiving bottle was connected. 100 mL of petroleum ether was added to the upper end of the condenser tube, and heated to reflux extraction for 2 hours. A ground glass rod was used to take 1 drop of the extract, and no oil spots were observed. The extraction was complete. The receiving bottle was removed, and the solvent was recovered by rotary evaporation (80°C, 60 rpm, vacuum degree 0.07-0.08 MPa, about 10 minutes). The yield was calculated by cooling and weighing. The same conditions were repeated 3 times to obtain the average value.

[0054] Determination of eucommia rubber content: 5 g of eucommia peel powder was accurately weighed, added with 100 ml of petroleum ether solvent in a Soxhlet extractor, heated in a water bath at 80-85 °C for 2 h, then filtered under heat with a 120 mesh stainless steel screen, separated the residue, and collected the eucommia rubber-dissolved filtrate in a stoppered flask. After the eucommia rubber-dissolved filtrate was cooled with water, it was stored in a low-temperature (-20 °C) refrigerator for about 1 h, filtered with a 120 mesh stainless steel screen to obtain eucommia rubber and eucommia rubber-removed filtrate. The filtrate was returned to the residue and re-heated for 1-2 h to completely dissolve the residual eucommia rubber, under the same conditions and operations as the first time. The recovered eucommia rubber was combined with the first eucommia rubber to obtain the product, and its total mass was accurately weighed.

[0055] As can be seen from Table 2, the fruits of eucommia were significantly increased after the Bacillus aryabhattai FS109 root irrigation treatment. The longitudinal diameter of the fruits was 35.98 mm, which was 17.3% higher than that of the untreated, and the hundred-grain dry fruit weight was 8.92 g, which was 23.2% higher than the 7.24 g of the control.

[0056] Table 2 Effect of FS109 treatment on the growth of eucommia fruits

[0057]

[0058]

[0059] The contents of chlorogenic acid and aucubin in the leaves, and the contents of volatile oil and eucommia rubber in the fruits were determined. As can be seen from Table 3, the contents of chlorogenic acid and aucubin in the leaves were increased by 30.3% and 23.0% respectively after the Bacillus aryabhattai FS109 root irrigation treatment, and the difference was significant. However, the contents of volatile oil and eucommia rubber in the fruits did not show significant changes.

[0060] Table 3 Effect of FS109 on the important components of eucommia leaves and fruits

[0061]

[0062] 3 Cultivation experiment of forsythia suspensa

[0063] 3.1 Fertilization

[0064] Potted seedlings: The experiment was carried out in the College of Pharmacy, Henan University. In late October, forsythia suspensa seedlings which were cut in August were transplanted into nutrient pots, and Bacillus aryabhattai was inoculated by root irrigation at a concentration of 1 x 10 8 cfu.mL -1 -1, with a inoculation amount of 20 mL / plant, 10 plants per treatment, 3 repeats, and water as the control. The forsythia suspensa seedlings were placed in a constant-temperature greenhouse with an illumination intensity of 2500 lux and a day length of 12 hours / day.

[0065] Field plants: The experiment was carried out in Lingbao City Sanmenxia Deme Co., Ltd. Fu Ling Chuan planting base. In April 2024, three-year-old fu ling chuan plants were selected, and the variety was Lingqiao No. 1. Two trenches were opened under the root crown of the plants, with a depth of 15 cm, and 50 mL of bacterial suspension was poured in. Each treatment was 10 plants, and each treatment was repeated 3 times. The control group was treated with water.

[0066] 3.2 Chlorophyll content determination

[0067] Potted seedlings: At 50 days, 10 leaves of each plant were collected and thoroughly mixed as a mixed sample for use.

[0068] Field plants: On August 15, 10 leaves of each plant were collected from the 4th to 5th nodes of the middle branches from top to bottom, and thoroughly mixed as a mixed sample, placed in a foam box with ice bags, and brought back to the laboratory for use.

[0069] Chlorophyll a and chlorophyll b content determination method: Chlorophyll extraction was carried out by anhydrous ethanol extraction method, avoiding the edge and veins of the leaves, and cutting the leaves into 2mm wide silk with scissors. Dark soak for 12h until white, use purple spectrophotometer to determine chlorophyll a and chlorophyll b at 645nm and 663nm respectively, and chlorophyll content is calculated by Arnon method.

[0070] From Figure 2 and Figure 3 It can be seen that after 30 days of Bacillus aryabhattai FS109 root irrigation treatment, the chlorophyll a and chlorophyll b content in the leaves of fu ling chuan seedlings and field plants increased significantly. Chlorophyll plays an important role in light energy absorption, transmission and conversion, and the increase of chlorophyll content can increase the photosynthetic rate of plants, thereby promoting the synthesis of plant organic matter and promoting plant growth.

[0071] 3.3 Plant growth index determination

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

[0073] Field plants: On August 15, the fruit ripening period, the branch growth was determined.

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

[0075] Table 4 Effect of FS109 on the growth of forsythia seedlings

[0076]

[0077] As can be seen from Table 5, compared with the water control group, after 30 days of Bacillus aryabhattai FS109 root irrigation treatment, the leaf length and width of forsythia field plants did not show significant growth. However, the trunk growth was significantly different from the water control. More importantly, the fruit length increased by 10.3% and the hundred-grain weight increased by 11.4% compared with the water control.

[0078] 3.4 Fruit growth index determination

[0079] On August 15, the fruit ripening period, fruits were collected and taken back to the laboratory for standby. The size and hundred-grain weight of forsythia fruits were determined.

[0080] Table 5 Effect of FS109 treatment on the growth of forsythia field plants and fruit size

[0081]

[0082] 3.5 Active ingredient content determination

[0083] Pot seedlings: Take the leaves of forsythia seedlings, and determine the contents of forsythiin and forsythoside A: accurately weigh 2 mg of forsythiin or forsythoside A reference substance, add 2 mL of methanol to prepare 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, and place in a conical flask with a stopper. Accurately add 25 mL of methanol, weigh, ultrasonic treat (power 250 W, frequency 40 kHz) for 25 minutes, cool, re-weigh, make up the weight loss with methanol, shake well, filter, and take the filtrate. Determine the content of forsythiin. Take about 0.5 g of the powder (passed through a No. 5 sieve), accurately weigh, place in a conical flask with a stopper, accurately add 15 mL of 70% methanol, tightly stopper, weigh, ultrasonic treat (power 250 W, frequency 40 kHz) for 30 minutes, cool, re-weigh, make up the weight loss with 70% methanol, shake well, filter, and take the filtrate. Determine the content of forsythoside A. The experimental data are subjected to significant difference test by T test of SPSS.

[0084] Field: Take forsythia fruits, and determine the contents of forsythiin and forsythoside A.

[0085] Experimental data analysis

[0086] Significant difference test by T test of SPSS.

[0087] As can be seen from Table 6, compared with the water control group, the internal quality of forsythia fruit is significantly improved. After Bacillus aryabhattai FS109 root irrigation treatment, the content of forsythoside reaches 0.29%, while the water control group is 0.22%, and the increase is 31.8%; the content of forsythoside A reaches 10.7%, while the water control group is 8.1%, and the increase is 32.1. The content of volatile oil does not change significantly. The problem of off-specification forsythoside in old kuo is a difficult problem for traditional Chinese medicine processing enterprises. The 2020 edition of Chinese Pharmacopoeia stipulates that the content of forsythoside shall not be less than 0.15%. With the extension of time, forsythoside in forsythia will degrade. The Bacillus aryabhattai FS109 of the present patent can increase forsythoside in green kuo by 31.8%, which can provide guarantee for producing qualified old kuo in the later period.

[0088] Table 6 Effect of FS109 treatment on active ingredients in fruit of forsythia field plants

[0089]

[0090] In summary, the strain Bacillus aryabhattai FS109 with antibiosis effect used in eucommia ulmoides and forsythia can promote plant photosynthesis, promote the growth of root system and aboveground part, and increase the content of active ingredients in medicinal parts, and has 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 only for the preferred embodiments of the present application, and not for limiting the present application, although the present application is described in detail with reference to the foregoing embodiments, for those skilled in the art, it still can modify the technical solutions recorded in the foregoing embodiments, or make equivalent replacement for part of the technical features. Any modification, equivalent replacement, improvement, etc. within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. Bacillus aryabhattai FS109, designated as Bacillus aryabhattai Bacillus aryabhattai FS109, designated as Bacillus aryabhattai FS109, designated as Bacillus aryabhattai FS109, designated as Bacillus aryabhattai FS109, designated as Bacillus aryabhattai FS109, designated as Bacillus aryabhattai FS109, designated as Bacillus aryabhattai FS109, designated as Bacillus aryabhattai FS109, designated as Bacillus aryabhattai FS109, designated as Bacillus aryabhattai FS109, designated as Bacillus aryabhattai FS109, designated as B 2. The Bacillus aryabhattai FS109 culture method according to claim 1, characterized by, The culture medium is lactose 1.7g, beef infusion powder 2.4g, MgSO4 1.5g, agar 20g, water 1000ml, pH7.

3. The Bacillus aryabhattai FS109 culture method according to claim 2, characterized by, The culture conditions were temperature 35 ℃, rotation speed 180 r·min -1 .

4. Use of the Bacillus aryabhattai FS109 of claim 1 in promoting photosynthesis of Eucommia ulmoides or Forsythia suspensa.

5. Use of the Bacillus aryabhattai FS109 of claim 1 in promoting growth of Eucommia ulmoides or Forsythia suspensa.

6. Use of the Bacillus aryabhattai FS109 of claim 1 in promoting increase of active ingredients of Eucommia ulmoides or Forsythia suspensa, wherein the active ingredients are any one or more of chlorogenic acid, onjisaponin, forsythoside or forsythoside A.

7. Use according to claim 6, characterized in that, The Bacillus aryabhattai FS109 bacterial solution is poured into the root cap of Eucommia ulmoides or Forsythia suspensa.

8. Use according to claim 7, characterized in that, The concentration of Bacillus aryabhattai FS109 bacterial solution was 1 x 10 8 cfu·mL -1 The amount was 20-100 mL.

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