A composite microbial agent and application thereof in quality improvement of fritillaria

Abstract

The application belongs to the field of microorganisms and biotechnology, and particularly relates to a compound microbial agent and application thereof in quality improvement of Fritillariae Thunbergii Miq. The compound microbial agent comprises microbacterium DGWGJ-1, microbacterium BM-1 and microbacterium BM-2, and further comprises azotobacter GSICC30112. The compound microbial agent not only retains the functional characteristics of the original microbial agent, but also significantly improves the total alkaloid content of medicinal components of 2-year-old and 4-year-old Fritillariae Thunbergii Miq., promotes the accumulation of jasmonic acid, and has a significant promoting effect on the quality improvement of Fritillariae Thunbergii Miq. The compound microbial agent can be used for preparing Fritillariae Thunbergii Miq. quality improvement regulator, quality improvement special fertilizer and the like, and has a broad application prospect.

Description

Technical Field

[0001] This invention belongs to the field of microbiology and biotechnology, specifically relating to a compound microbial agent and its application in improving the efficacy and quality of Fritillaria cirrhosa. Background Technology

[0002] Fritillaria cirrhosa, a perennial herb belonging to the genus Fritillaria in the family Liliaceae, is a "National Class II Key Protected Wild Plant." As one of the original plants of the precious medicinal herb "Chuanbei," its bulb has the effects of clearing heat and moistening the lungs, resolving phlegm and relieving cough. Wild resources of Fritillaria cirrhosa are mainly distributed in shrublands or grasslands at altitudes of 2,800–4,400 meters in Wudu, Zhouqu, Maqu, Minxian, and Yuzhong areas of Gansu Province. It prefers cool, moist, and fertile soil, and dislikes high temperatures and direct sunlight, exhibiting stringent environmental requirements. Because traditional artificial cultivation methods require 4-5 years from sowing to the formation of commercial medicinal materials, which is a long cycle and costly, the development of efficacy-enhancing microbial agents to achieve rapid and high-quality production is imperative.

[0003] "Quality first, with yield as a secondary consideration" is the basic principle of medicinal herb cultivation. In recent years, ecological cultivation has become the main model for the cultivation and production of Chinese medicinal herbs. The requirements for the production environment and inputs in ecological agriculture for Chinese medicinal herbs are the same as those for organic agriculture. During cultivation, the "six no's" principle is adhered to: no use of chemically synthesized fertilizers, pesticides, agricultural films, herbicides, hormones, or genetically modified seeds. Cultivation should comprehensively utilize technologies such as three-dimensional planting, biological control of crop diseases and pests, facility agriculture, high-value recycling of organic fertilizers and crop straw, modern microbial technology, and resistant variety breeding. The safety requirements of the products must meet organic product standards (Guo Lanping et al., Latest Developments and Prospects of Ecological Agriculture for Chinese Medicinal Herbs).

[0004] The "adversity effect" of traditional Chinese medicinal herbs indicates that the suitable habitat required for the accumulation of secondary metabolites in plants may not be consistent with, or even opposite to, the suitable habitat for their growth and development. When medicinal plants face certain specific stresses over a long period, they may defend against these stresses by synthesizing and accumulating specific secondary metabolites. Therefore, moderate environmental stress may induce the accumulation of secondary metabolites (Huang Luqi et al., Accumulation of Secondary Metabolites under Environmental Stress and the Formation of Traditional Chinese Medicinal Herbs). When plants are subjected to environmental stress, they activate the expression of stress-related endogenous hormones such as salicylic acid and jasmonic acid. Studies have shown that the increase of endogenous SA and JA in belladonna can increase the content of secondary metabolites hyoscyamine and scopolamine (Shan Yusi et al., Effects of Exogenous Methyl Jasmonate and Salicylic Acid on Physiological Characteristics and Secondary Metabolism of Belladonna under Salt Stress); spraying with 200 μmol·L... -1Exogenous methyl jasmonate promotes the growth of fresh and dry weight of Uncaria rhynchophylla seedlings and the accumulation of alkaloids (Lu Xingxing et al., Effects of methyl jasmonate on Uncaria rhynchophylla alkaloid synthesis); Rehmannia glutinosa, after being infected by endophytic fungi, increases JA content by inducing JA-related synthases, activates the expression of plant stress resistance genes, and accelerates the synthesis of secondary metabolites such as catalpol and verbascoside (Peng Shuping et al., Cloning and expression analysis of two key genes for jasmonate synthesis in Rehmannia glutinosa in response to endophytic fungal infection); Exogenous JA can induce enhanced activity of protective enzymes in Chrysanthemum morifolium leaves and increase the content of endogenous JA, flavonoids, and phenolic compounds (Fan Jie et al., Effects of methyl jasmonate on chrysanthemum aphid resistance).

[0005] Unlike other cash crops, the economic value of fritillaria does not depend on its yield, but primarily on its medicinal efficacy. Therefore, smaller fritillaria bulbs are more valuable, with the highest-valued being the Songbei variety, which has a crescent-shaped shape and a diameter of 0.5-0.8 cm; followed by the Qingbei variety, which has two equally sized lobes that interlock, with a diameter of 0.8-1.5 cm. However, the application of existing products used to promote growth (auxin, cytokinin, gibberellin) and increase yield to fritillaria can actually reduce its medicinal efficacy and quality, thus affecting its commercial value. Studies on improving the quality of Fritillaria cirrhosa have found that the alkaloid content of three-year-old Gansu Fritillaria cirrhosa currently under cultivation is 0.044%, which decreases to 0.052% after treatment with nano-iron, representing an increase of 18.2%, but the absolute content barely meets the pharmacopoeia standard. In contrast, the alkaloid content of the blank control of four-year-old Fritillaria cirrhosa is 0.163%, which decreases to 0.128% after treatment with nano-iron, a reduction of 21.5% (Yang Tao et al., Effects of Nano-Iron and Melatonin on Yield and Quality of Gansu Fritillaria cirrhosa under Domestication Cultivation Conditions). This indicates that the effect of nano-iron on the quality of Fritillaria cirrhosa from different years is not stable.

[0006] Therefore, based on the "adversity effect" of the formation of the quality of Chinese medicinal materials, this invention provides a compound microbial agent that can effectively promote the accumulation of endogenous jasmonic acid in fritillaria and increase the content of alkaloids, and has a significant quality improvement effect without reducing the yield. Summary of the Invention

[0007] To address the aforementioned technical problems, the present invention aims to provide a compound microbial agent and its application in improving the efficacy and quality of Fritillaria cirrhosa. After spraying the plant with the compound microbial agent, the content of the stress-related hormone jasmonic acid significantly increases, thereby increasing the alkaloid content in Fritillaria cirrhosa and improving its quality. Specifically, it includes the following:

[0008] In a first aspect, the present invention provides a composite microbial agent, the composite microbial agent comprising Microbacterium, Microbacterium tumefaciens, and Microbacterium oxyphylla.

[0009] Preferably, the microbacterium is microbacterium DGWGJ-1, with a 16S rDNA sequence as shown in SEQ ID NO.1 and NCBI number MW981366; the microbacterium var. microphylla is microbacterium var. microphylla BM-1, with a 16S rDNA sequence as shown in SEQ ID NO.2 and NCBI number MW981364; and the microbacterium oxidans is microbacterium oxidans BM-2, with a 16S rDNA sequence as shown in SEQ ID NO.3 and NCBI number MW981365.

[0010] Preferably, the compound microbial agent is prepared by mixing the above-mentioned strains in equal proportions.

[0011] Preferably, the bacterial agent contains 10 CFU of the microbial strain. 6 ~10 9 / ml.

[0012] Preferably, the compound microbial agent also includes *Azotobacter chrysophyte*.

[0013] Preferably, the azotocinobacter is Azotocinobacter chrysozoata GSICC30112, and its 16S rDNA sequence is shown in SEQ ID NO.4.

[0014] Preferably, the compound microbial agent is prepared by mixing the above-mentioned strains in equal proportions.

[0015] Preferably, the bacterial agent contains 10 CFU of the microbial strain. 6 ~10 9 / ml.

[0016] Secondly, the present invention provides an application of the composite microbial agent described in the first aspect above in the preparation of plant quality regulators.

[0017] Preferably, the plant is Fritillaria cirrhosa.

[0018] Fourthly, the present invention provides an application of the compound microbial agent described in the first aspect above in the preparation of a special fertilizer for improving plant quality.

[0019] Preferably, the plant is Fritillaria cirrhosa.

[0020] Preferably, the compound microbial agent can significantly increase the alkaloid content of plants.

[0021] Preferably, the compound microbial agent can significantly increase the jasmonic acid content in plants.

[0022] The beneficial effects of this invention are as follows: Firstly, this invention provides a compound microbial agent, which includes Microbacterium DGWGJ-1, Microbacterium BM-1, and Microbacterium BM-2; the compound microbial agent also includes Azotobacter chrysophyte GSICC30112; the compound microbial agent of this invention can significantly increase the content of alkaloids, the active medicinal component of Fritillaria, and promote the accumulation of jasmonic acid, thus having a significant quality-improving effect, increasing the commercial value of Fritillaria, and having significant economic significance. Detailed Implementation

[0023] The present invention will be further described below with reference to specific embodiments. However, the scope of protection of the present invention is not limited to the following embodiments. Any modifications or substitutions made to the methods, steps or conditions of the present invention without departing from the spirit and substance of the present invention shall fall within the scope of the present invention.

[0024] The test plants described in the following examples are *Fritillaria cirrhosa* from Gansu Province. The test site is located in Habancha Village, Mapo Township, Yuzhong County, Lanzhou City, Gansu Province, at an altitude of 2600 meters. All reagents and culture media used in the experiment were chemically pure.

[0025] The microorganisms used in the following examples are: Microbacterium DGWGJ-1, 16S rDNA sequence as shown in SEQ ID NO.1, NCBI number MW981366, deposited at the China General Microbiological Culture Collection Center (CGMCC NO18608); Microbacterium BM-1, 16S rDNA sequence as shown in SEQ ID NO.2; Microbacterium oxidans BM-2, 16S rDNA sequence as shown in SEQ ID NO.3; and Azotobacter chrysopraserum GSICC30112, 16S rDNA sequence as shown in SEQ ID NO.4, all purchased from the Gansu Branch of the China Industrial Microbiological Culture Collection Center.

[0026] *Bacillus belye* 4C5 (NCBI No. MW989745), *Bacillus amyloliquefaciens* 5C5 (NCBI No. MW981362), *Bacillus subtilis* 5C1 (NCBI No. MW981361), and *Brugia hygroscopica* AL1 (NCBI No. MW981363) were isolated from medicinal plants by members of the research group and were deposited and purchased from the Gansu Branch of the China Industrial Microbial Culture Collection Center.

[0027] Culture medium components:

[0028] PDB medium: 200 g / L potato, 20 g / L glucose, natural pH.

[0029] Experimental group Microbacterium compound inoculum (T1): Microbacterium DGWGJ-1, Microbacterium BM-1, and Microbacterium BM-2 cultured in PDB medium were mixed in equal proportions and adjusted to a total bacterial concentration of 1×10⁻⁶. 7 CFU·mL -1 ;

[0030] Experimental group single bacterial inoculum (T2): Azotobacter chrysophagus GSICC30112 was cultured in PDB medium and adjusted to a bacterial concentration of 1×10⁻⁶. 7 CFU·mL -1 ;

[0031] The experimental group's compound bacterial agent (T3) was prepared by mixing equal proportions of *Microbacterium glutamicum* DGWGJ-1, *Microbacterium miltiorrhiza* BM-1, *Microbacterium oxysporum* BM-2, and *Azotobacter chrysophagus* GSICC30112 cultured in PDB medium, and adjusting the total bacterial concentration to 1×10⁻⁶. 7 CFU·mL -1 ;

[0032] Positive control group compound bacterial agent (T4): *Bacillus belyceae* 4C5 (NCBI No. MW989745), *Bacillus amyloliquefaciens* 5C5 (NCBI No. MW981362), *Bacillus subtilis* 5C1 (NCBI No. MW981361), and *Brugia hygroscopica* AL1 (NCBI No. MW981363) cultured in PDB medium were mixed in equal proportions and adjusted to a total bacterial concentration of 1×10⁻⁶. 7 CFU·mL -1 ;

[0033] Negative control group (CK): PDB medium without bacteria.

[0034] The total alkaloid content was determined according to the methods described in the 2020 edition of the Chinese Pharmacopoeia (National Pharmacopoeia Commission. Pharmacopoeia of the People's Republic of China: Part I).

[0035] Example 1: Effects of different treatments on the quality and yield of 4-year-old fritillaria bulbs

[0036] The experiment employed a single-factor completely randomized design, with five treatments: T1, T2, T3, T4, and CK. Different bacterial strains were subjected to different treatments at 28℃ and 200 rpm. -1 Under the specified conditions, cultured in PDB medium with shaking for 2 days, adjusted to the same OD value, then reconstituted in equal proportions and adjusted to 1×10⁻⁶ with sterile water. 7 CFU·mL -1 The application volume was the same for all treatments. Each treatment was replicated three times, with a plot area of ​​5m². 2After seedlings emerge in early April, different treatments are foliar sprayed once every 3 weeks for a total of 4 times. After the above-ground parts wither in mid-July, 300 plants are dug up from each plot, rinsed with running water, and the roots and surface soil are removed. After being air-dried, they are weighed and the total alkaloids and jasmonic acid content are tested.

[0037] The results of total alkaloids and yield testing are shown in Table 1.

[0038] Table 1. Results of determination of total alkaloid content and yield of Fritillaria cirrhosa after different treatments ( n=3)

[0039]

[0040] Note: Each column of capital letters represents multiple comparisons at the highly significant level (P<0.01).

[0041] The results showed that the total alkaloid content in experimental groups T1, T2, and T3 was increased by 1.31 times, 1.49 times, and 1.57 times, respectively, compared with the negative control CK; while the total alkaloid content in the positive control group T4 was decreased by 27% compared with the negative control CK. This indicates that the compound microbial agent described in this invention significantly increased the content of the medicinal alkaloids in 4-year-old Fritillaria cirrhosa, demonstrating a significant quality-improving effect. Simultaneously, experimental groups T1, T2, and T3 had no significant impact on the yield of 4-year-old Fritillaria cirrhosa compared with the negative control CK. That is, the compound microbial agent described in this invention significantly promoted the content of the medicinal alkaloids in 4-year-old Fritillaria cirrhosa without affecting the yield, demonstrating a significant quality-improving effect.

[0042] Seven days after the fourth treatment, leaves from four-year-old Fritillaria thunbergii plants were harvested, and the contents of hormones (jasmonic acid, salicylic acid, gibberellins 3, 4, and 7, cytokinins, and auxins) were determined according to the method described in the literature (Yang Tao, Effects of nano iron and melatonin on the yield and quality of Fritillaria thunbergii under domestication cultivation conditions in Gansu). Ten Fritillaria thunbergii plants were randomly selected from each plot, and leaves were harvested and mixed for testing.

[0043] The hormone test results are shown in Table 2.

[0044] Table 2. Results of Fritillaria hormone content after different treatments (ng / g) n=3)

[0045]

[0046] Note: Each column of capital letters represents multiple comparisons at the highly significant level (P<0.01).

[0047] The results showed that the jasmonic acid content in experimental groups T1, T2, and T3 was increased by 40.9%, 54%, and 50.5% respectively compared to the negative control CK; while the jasmonic acid content in the positive control group T4 was decreased by 81% compared to the negative control CK. This indicates that the compound microbial agent described in this invention significantly promoted the accumulation of jasmonic acid in 4-year-old Fritillaria cirrhosa, demonstrating a significant quality-improving effect. In the positive control group T4, the levels of salicylic acid, gibberellin, cytokinin, and auxin increased by 49.1%, 6.24 times, 5.75 times, and 1.61 times respectively compared to the negative control CK, leading to a significant increase in yield and impacting the commercial value of Fritillaria cirrhosa; while the levels of salicylic acid, gibberellin, cytokinin, and auxin in experimental groups T1, T2, and T3 showed no significant changes compared to the negative control CK, having no significant impact on yield.

[0048] Example 2: Effects of different treatments on the quality of 2-year-old fritillaria bulbs

[0049] The experiment employed a single-factor completely randomized design, with four treatments: T1, T2, T3, and CK. Different strains were subjected to different treatments at 28℃ and 200 rpm. -1 Under the specified conditions, cultured in PDB medium with shaking for 2 days, adjusted to the same OD value, then reconstituted in equal proportions and adjusted to 1×10⁻⁶ with sterile water. 7 CFU·mL -1 The application volume was the same for all treatments. Each treatment was replicated three times, with a plot area of ​​5m². 2 After seedlings emerge in early April, different treatments are foliar sprayed once every 3 weeks for a total of 4 times. After the above-ground parts wither in mid-July, 300 plants are dug up from each plot, rinsed with running water, and the roots and surface soil are removed. After air drying, the total alkaloid content is tested.

[0050] The results of the total alkaloid content test are shown in Table 3.

[0051] Table 3. Results of determination of total alkaloid content in Fritillaria cirrhosa after different treatments ( n=3)

[0052]

[0053] Note: Uppercase letters indicate multiple comparisons at the highly significant level (P<0.01).

[0054] The results showed that the total alkaloid content in experimental groups T1, T2, and T3 was increased by 52.4%, 57.1%, and 80.9% respectively compared with the negative control CK. This indicates that the compound microbial agent described in this invention also significantly increased the content of alkaloids, the active medicinal component, in 2-year-old Fritillaria cirrhosa, and has a significant quality-improving effect.

[0055] In summary, the compound microbial agent described in this invention can significantly increase the content of alkaloids, the active medicinal component of Fritillaria, and promote the accumulation of jasmonic acid, thus exhibiting significant quality-improving effects and significantly enhancing the commercial value of Fritillaria, which is of great economic significance. sequence list <110> Gansu Academy of Sciences Institute of Biology <120> A compound microbial agent and its application in the extraction of Fritillaria cirrhosa. <160> 4 <170> SIPOSequenceListing 1.0 <210> 1 <211> 1013 <212> DNA <213> Microbacterium <400> 1 gaaatacctt cgacgctccc tcccacaagg ggttaggcca ccggcttcag gtgttaccga 60 ctttcatgac ttgacgggcg gtgtgtacaa gacccgggaa cgtattcacc gcagcgttgc 120 tgatctgcga ttactagcga ctccgacttc atgaggtcga gttgcagacc tcaatccgaa 180 ctgggaccgg ctttttggga ttcgctccac cttacggtat cgcagccctt tgtaccggcc 240 attgtagcat gcgtgaagcc caagacataa ggggcatgat gatttgacgt catccccacc 300 ttcctccgag ttgaccccgg cagtatccca tgagttccca ccattacgtg ctggcaacat 360 agaacgaggg ttgcgctcgt tgcgggactt aacccaacat ctcacgacac gagctgacga 420 caaccatgca ccacctgttc acgagtgtcc aaagagttga ccatttctgg cccgttctcg 480 tgtatgtcaa gccttggtaa ggttcttcgc gttgcatcga attaatccgc atgctccgcc 540 gcttgtgcgg gtccccgtca attcctttga gttttagcct tgcggccgta ctccccaggc 600 ggggaactta atgcgttagc tgcgtcacgg aatccgtgga aaggacccca caactagttc 660 ccaacgttta cggggtggac taccagggta tctaagcctg tttgctcccc accctttcgc 720 tcctcagcgt cagttacggc ccagagatct gccttcgcca tcggtgttcc tcctgatatc 780 tgcgcattcc accgctacac caggaattcc aatctcccct accgcactct agtctgcccg 840 tacccactgc aggctggagg ttgagcctcc agatttcaca gcagacgcga cagaccgcct 900 acgagctctt tacgcccaat aattccggat aacgcttgcg ccctacgtat taccgcggct 960 gctggcacgt aattagccgg ggctttttct gcagggaccg tcctctcgct tct 1013 <210> 2 <211> 857 <212> DNA <213> Microbacterium <400> 2 gttacggatg ttacctgcaa gtcgaacggt gaacacggag cttgctctgt gggatcagtg 60 gcgaacgggt gagtaacacg tgagcaacct gcccctgact ctgggataag cgctggaaac 120 ggcgtctaat actggatacg agtagcgatc gcatggtcag ctactggaaa gattttttgg 180 ttggggatgg gctcgcggcc tatcagcttg ttggtgaggt aatggctcac caaggcgtcg 240 acgggtagcc ggcctgagag ggtgaccggc cacactggga ctgagacacg gcccagactc 300 ctacgggagg cagcagtggg gaatattgca caatgggcgg aagcctgatg cagcaacgcc 360 gcgtgaggga tgacggcctt cgggttgtaa acctctttta gcagggaaga agcgaaagtg 420 acggtacctg cagaaaaagc gccggctaac tacgtgccag cagccgcggt aatacgtagg 480 gcgcaagcgt tatccggaat tattgggcgt aaagagctcg taggcggttt gtcgcgtctg 540 ctgtgaaatc ccgaggctca acctcgggcc tgcagtgggt acgggcagac tagagtgcgg 600 taggggagat tggaattcct ggtgtagcgg tggaatgcgc agatatcagg aggaacaccg 660 atggcgaagg cagatctctg ggccgtaact gacgctgagg agcgaaaggg tggggagcaa 720 acaggcttag ataccctggt agtccacccc gtaaacgttg ggaactagtt gtggggtcca 780 ttccacggat tccgtgacgc agctaacgca ttaagttccc cgcctgggga gtacggccgc 840 aaggctaaaa ctcaaag 857 <210> 3 <211> 1004 <212> DNA <213> Microbacterium <400> 3 gtataccttc gacggctccc tccacaaggg ttaggccacc ggcttcaggt gttaccgact 60 ttcatgactt gacgggcggt gtgtacaaga cccgggaacg tattcaccgc agcgttgctg 120 atctgcgatt actagcgact ccgacttcat gaggtcgagt tgcagacctc aatccgaact 180 gggaccggct ttttgggatt cgctccacct cgcggtattg cagccctttg taccggccat 240 tgtagcatgc gtgaagccca agacataagg ggcatgatga tttgacgtca tccccacctt 300 cctccgagtt gaccccggca gtatcccatg agttcccacc attacgtgct ggcaacatag 360 aacgagggtt gcgctcgttg cgggacttaa cccaacatct cacgacacga gctgacgaca 420 accatgcacc acctgtttac gagtgtccaa agagttgacc atttctggcc cgttctcgta 480 tatgtcaagc cttggtaagg ttcttcgcgt tgcatcgaat taatccgcat gctccgccgc 540 ttgtgcgggt ccccgtcaat tcctttgagt tttagccttg cggccgtact ccccaggcgg 600 ggaacttaat gcgttagctg cgtcacggaa tccgtggaat ggaccccaca actagttccc 660 aacgtttacg gggtggacta ccagggtatc taagcctgtt tgctccccac cctttcgctc 720 ctcagcgtca gttacggccc agagatctgc cttcgccatc ggtgttcctc ctgatatctg 780 cgcattccac cgctacacca ggaattccaa tctcccctac cgcactctag tctgcccgta 840 cccactgcag gccggaggtt gagcctccgg atttcacagc agacgcgaca aaccgcctac 900 gagctcttta cgcccaataa ttccggataa cgcttgcgcc ctacgtatta ccgcggctgc 960 tggcacgtag ttagccggcg ctttttctgc aggtaccgtc cttt 1004 <210> 4 <211> 1224 <212> DNA <213> Azotobacter chroococcum <400> 4 ggggtgcttg catcccgatt cagcggcgga cgggtgagta atgcctagga atctgcccga 60 tagtggggga caacgtttcg aaaggaacgc taataccgca tacgtcctac gggagaaagt 120 gggggctctt cggacctcac gctatcggat gagcctaggt cggattagct agttggtggg 180 gtaaaggctc accaaggcga cgatccgtaa ctggtctgag aggatgatca gtcacactgg 240 aactgagaca cggtccagac tcctacggga ggcagcagtg gggaatattg gacaatgggc 300 gaaagcctga tccagccatg ccgcgtgtgt gaagaaggtc ttcggattgt aaagcacttt 360 aagttgggag gaagggctgt aagcgaatac cttgcagttt tgacgttacc gacagaataa 420 gcaccggcta acttcgtgcc agcagccgcg gtaatacgaa gggtgcaagc gttaatcgga 480 attactgggc gtaaagcgcg cgtaggtggt ttggtaagtt ggatgtgaaa gccccgggct 540 caacctggga actgcatcca aaactgcctg actagagtac ggtagagggt ggtggaattt 600 cctgtgtagc ggtgaaatgc gtagatatag gaaggaacac cagtggcgaa ggcgaccacc 660 tggactgata ctgacactga ggtgcgaaag cgtggggagc aaacaggatt agataccctg 720 gtagtccacg ccgtaaacga tgtcgactag ccgttgggct ccttgagagc ttagtggcgc 780 agctaacgca ttaagtcgac cgcctggga gtacggccgc aaggttaaaa ctcaaatgaa 840 ttgacggggg cccgcacaag cggtggagca tgtggtttaa ttcgaagcaa cgcgaagaac 900 cttacctggc cttgacatgc tgagaacttt ccagagatgg attggtgcct tcgggaactc 960 agacacaggt gctgcatggc tgtcgtcagc tcgtgtcgtg agatgttggg ttaagtcccg 1020 taacgagcgc aacccttgtc cttagttaca gcacctcggg tgggcactct aaggagactg 1080 ccgtgaaaa ccggagaaa ggtggggatg acgtcaagtc atcatggccc ttacgggcag 1140 gcctacacac gtgctacaat ggtcggtaca gagggttgcc aagtcgcgag gcggagctaa 1200 tcccagaaaa ccgatcgtat tccg 1224

Claims

1. A compound microbial agent, characterized in that, The compound microbial agent includes Microbacterium DGWGJ-1, Microbacterium BM-1, and Microbacterium BM-2; the 16S rDNA sequence of Microbacterium DGWGJ-1 is shown in SEQ ID NO.1; the 16S rDNA sequence of Microbacterium BM-1 is shown in SEQ ID NO.2; and the 16S rDNA sequence of Microbacterium BM-2 is shown in SEQ ID NO.

3.

2. The compound microbial agent as described in claim 1, characterized in that, The compound microbial agent also includes Azotobacter chrysotrichum gSICC30112; the 16S rDNA sequence of Azotobacter chrysotrichum gSICC30112 is shown in SEQ ID NO.

4.

3. The application of the compound microbial agent as described in any one of claims 1-2 in the preparation of Fritillaria cirrhosa quality regulator.

4. The application of the compound microbial agent as described in any one of claims 1-2 in the preparation of special fertilizer for improving the quality of Fritillaria.

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