Marsinia sp. Strain for promoting growth of Aucaria kjankei and application of Marsinia sp. Strain
By isolating and screening out of the Arachite Root tumor of the Arachite Root tumor of the Arachite Root tumor, the problem of slow growth in the Arachite seedling stage of the Arachite is solved, significantly promoting its mycorrhizal formation and growth, and providing high-quality bacterial fertilizer resources to support its afforestation practice.
Patent Information
- Application Number
- CN202411779736.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-05
- Publication Date
- 2025-02-25
AI Technical Summary
Ken's Araucaria seedlings grow slowly and an effective method is needed to promote its growth.
Marseille strain E1-2h was isolated and screened from the Arachite Root tumor of Ken's Arachite Root tumor, and prepared into bacterial solution and injected into the soil around the Arachite Root system of Ken's Arachite Root system to promote mycorrhizal formation and plant growth.
Inoculation of Masai Cyperus E1-2h significantly promoted the formation of mycorrhizalis in Ken's Araucaria, increased underground dry weight, increased root surface area, and increased nodule tumor number per unit root surface area, improving plant growth.
Smart Images
Figure FT_1 
Figure FT_2 
Figure FT_3
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of microorganisms, and in particular relates to a strain of Massilia sp. capable of promoting the growth of Araucaria kennensis and application thereof. Background Art
[0002] Araucaria cunnighamii is an excellent tree species for both timber and greening in the Araucariaceae family. It is mainly distributed in the southeastern coastal areas of Oceania. It likes light and warm and humid climate. In its native place, the tree is 60-70 meters tall and has a breast diameter of more than 1 meter. This tree species is one of the five major park tree species in the world. The tree is tall, the trunk is straight and relatively round, the branches and leaves are stretched, the crown is spire-shaped, and the tree body is graceful. It is mainly cultivated as a bonsai and greening tree species abroad. A few countries have long managed it as a timber tree species, and its wood is widely used in construction, utensils and furniture industries. my country has made preliminary explorations in introduction experiments, cultivation techniques and propagation techniques. The results of the provenance and family test of Araucaria cunnighamii conducted by the Institute of Tropical Forestry, Chinese Academy of Forestry showed that the tree grew rapidly four years after planting, had strong adaptability, and beautiful forest appearance, and it is worth promoting planting in suitable areas in my country. At present, the market demand for Ken's Araucaria afforestation seedlings and ornamental plantings is also increasing. However, the tree species grows slowly in the seedling stage, and promoting the growth of Ken's Araucaria afforestation has become one of the important issues that need to be urgently addressed in the development of this tree species.
[0003] A large number of research results show that there are a large number of endophytic bacteria in plants that promote plant growth. Endophytic bacteria in plants have obvious promoting effects in increasing plant height, dry weight, increasing root weight and enhancing plant growth potential. The mechanisms by which endophytic bacteria promote plant growth include: directly promoting plant growth through biological nitrogen fixation or producing plant hormones; indirectly promoting plant growth by inducing host plants to produce plant hormones, improving plant utilization of minerals, changing host plants' sensitivity to harmful environmental conditions such as frost and harmful pathogens. The present invention isolated and screened a strain of Masai E1-2h that can promote the growth of Ken's Araucaria from the root nodules of Ken's Araucaria, enriching the Ken's Araucaria growth-promoting bacteria strain resource library and providing high-quality bacterial fertilizer germplasm resources for Ken's Araucaria afforestation practice. Summary of the invention
[0004] The invention aims to provide a strain of Massiliae for promoting the growth of Araucaria kennensis and application thereof.
[0005] To achieve the above object, the present invention adopts the following technical solution: A Massiliaspis strain that promotes the growth of Araucaria kennensis, the strain is named Massiliaspis (Massiliasp.) E1-2h, and was deposited in the General Microbiology Center of the China Culture Collection Administration on September 18, 2024, with the deposit number CGMCC No. 31989, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.
[0006] A microbial agent for promoting the growth of Araucaria kennellii, wherein the microbial agent contains the Massillaria strain according to claim 1.
[0007] Use of the above-mentioned Massilia strain or microbial agent in promoting the growth of Araucaria kennellii; Furthermore, the promoting the growth of Araucaria kennellii includes promoting the formation of mycorrhiza of Araucaria kennellii, increasing the dry weight of the underground part of Araucaria kennellii, increasing the root surface area of Araucaria kennellii, and increasing the number of nodules per unit root surface area of Araucaria kennellii.
[0008] A method for promoting the growth of Araucaria kennellii, comprising preparing the above-mentioned Massilia strain into a bacterial liquid and injecting the liquid into the soil around the root system of Araucaria kennellii; Furthermore, the promoting the growth of Araucaria kennellii includes promoting the formation of mycorrhiza of Araucaria kennellii, increasing the dry weight of the underground part of Araucaria kennellii, increasing the root surface area of Araucaria kennellii, and increasing the number of nodules per unit root surface area of Araucaria kennellii.
[0009] The significant advantages of the present invention are: The present invention separates and selects a strain of Masseella E1-2h that can promote the growth of Araucaria kennieri from the root nodules of Araucaria kennieri. The test results show that inoculation of Masseella E1-2h can promote the formation of Araucaria kennieri mycorrhiza, increase the underground dry weight of Araucaria kennieri, increase the root surface area of Araucaria kennieri, and increase the number of nodules per unit root surface area of Araucaria kennieri. The present invention enriches the Araucaria kennieri growth-promoting bacteria strain resource library and provides high-quality bacterial fertilizer germplasm resources for the Araucaria kennieri afforestation practice. BRIEF DESCRIPTION OF THE DRAWINGS
[0010] Figure 1 :Phylogenetic tree of Massillarism E1-2h constructed based on 16S rDNA gene.
[0011] Figure 2 : The whole plant morphology of Araucaria kennensis inoculated with live bacteria solution of Masseella E1-2h and the enlarged image of the mycorrhiza at a certain point of the root.
[0012] Figure 3 : The whole plant morphology of Araucaria kennensis inoculated with the inactivated bacterial solution of Masseille E1-2h and the enlarged image of the mycorrhiza at a certain point in the root.
[0013] Figure 4: Dry weight of underground part (g), dry weight of aboveground part (g), and root surface area (cm) of Araucaria kennellii inoculated with live bacteria solution of Masseille E1-2h and inactivated bacteria solution of Masseille E1-2h 2 ) and the number of nodules per unit root surface area. DETAILED DESCRIPTION
[0014] In order to make the contents of the present invention easier to understand, the technical solution of the present invention is further described below in conjunction with specific implementation methods, but the present invention is not limited thereto.
[0015] The YMB liquid culture medium of the present invention is composed of: 10 g / L mannitol, 0.4 g / L yeast powder, 0.1 g / L NaCl, 0.2 g / L MgSO4·7H2O, and 0.5 g / L K2HPO4; the YMA solid culture medium is composed of: 10 g / L mannitol, 3 g / L yeast powder, 0.1 g / L NaCl, 0.2 g / L MgSO4·7H2O, 0.25 g / L K2HPO4, 0.25 g / L KH2PO4, and 15 g / L agar.
[0016] Example 1: Isolation and identification of endophytic bacteria in root nodules of Araucaria kennellii Collect the plants of Araucaria kennelii, rinse the roots under the tap to remove the soil and impurities on the surface, cut the nodules with sterilized scissors, and clean and disinfect them. The specific method of cleaning and disinfection is: clean the surface with running water → wash once with sterile water → soak in 75% ethanol for 30s → rinse with sterile water 5 times → soak and disinfect in 10% sodium hypochlorite for 10min → rinse with sterile water 5 times. Place the cleaned and disinfected nodules on sterilized filter paper, cut into slices with a thickness of 1 to 2 mm with a sterile scalpel, transfer to YMB liquid culture medium, culture at 37°C, 120r / min shaking table for 1 to 3 days, and then repeatedly streak and purify on YMA solid culture medium to obtain strain E1-2h.
[0017] Morphological characteristics of strain E1-2h: The surface of the colony of strain E1-2h on YMA solid medium is smooth, sticky, and light yellow; it has no spores, and the microscopic morphology is short rod-shaped with flagella at the end. Physiological and biochemical characteristics of strain E1-2h: Gram-negative bacteria (G-), heterotrophic, aerobic, does not require light, and grows at 37°C.
[0018] 16S rDNA identification of strain E1-2h: strain E1-2h was inoculated into YMB liquid medium and cultured in a shaking incubator at 37°C and 120 r / min for 1-2 h. The obtained bacterial solution was used as a template to amplify the 16S rDNA fragment with the universal primers 27F (5'-AGAGTTTGATCMTGGCTCAG-3') and 1492R (5'-GGTTACCTTGTTACGACTT-3') of bacteria. The PCR amplification product was detected by 1.2% agarose gel electrophoresis and sent to a professional biological company for sequencing. The sequencing result (SEQ ID NO.1) was submitted to NCBI for sequence homology comparison, and the phylogenetic tree was constructed using bioinformatics software ( Figure 1 ) to determine the taxonomic status of the strains. The results showed that the closest relative to strain E1-2h was Massiliasp., with a similarity of 99.78%.
[0019] Based on comprehensive morphological characteristics, physiological and biochemical properties, and 16S rDNA gene sequence analysis, strain E1-2h was identified as Massiliasp., which was deposited in the General Microbiology Center of China Culture Collection on September 18, 2024, with the deposit number CGMCC No.31989, and the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.
[0020] Example 2: Growth-promoting effect of E1-2h of Massillaria serrata on Araucaria kennellii Preparation of live bacterial culture of Massillaria E1-2h: Massillaria E1-2h was inoculated into YMB liquid medium, activated in a shaker at 37°C and 120 r / min for 4 h, then transferred into a 50 mL centrifuge tube containing 30 mL YMB medium, and cultured in a shaker at 37°C and 120 r / min until the bacterial culture OD 600 The value was 0.8, the culture medium was removed by centrifugation, and then the suspension was resuspended with sterile ddH2O of the same volume as the culture medium to obtain the live bacterial solution of Masseille E1-2h. Preparation of the inactivated bacterial solution of Masseille E1-2h: The live bacterial solution of Masseille E1-2h was sterilized by ultraviolet irradiation for 15 minutes, and then sterilized at 121°C for 15 minutes to obtain the inactivated bacterial solution of Masseille E1-2h.
[0021] Kenneth's Araucaria plants were obtained by growing seeds of Kenneth's Araucaria in soil for 6 months, and then transferred to a long-day 28℃ greenhouse; one month after the plants were transferred to the greenhouse, live bacteria solution of Masseilles E1-2h and inactivated bacteria solution of Masseilles E1-2h were injected into the soil around the roots of a single Kenneth's Araucaria plant using a pipette, once a week, with 1mL of bacteria solution injected each time; after 4 consecutive months of inoculation, the growth of Kenneth's Araucaria was recorded. The results showed that ( Figure 2 , Figure 3 , Figure 4, Table 1), the aboveground growth of the Araucaria kennellii whose soil around the roots was injected with the live bacteria solution of Masseillium E1-2h was not much different from that of the Araucaria kennellii whose soil around the roots was injected with the inactivated bacteria solution of Masseillium E1-2h; but the mycorrhizal density of the Araucaria kennellii whose soil around the roots was injected with the live bacteria solution of Masseillium E1-2h was significantly higher than that of the Araucaria kennellii whose soil around the roots was injected with the inactivated bacteria solution of Masseillium E1-2h, and the number of mycorrhizae was relatively large; the underground dry weight, root surface area, and number of nodules per unit root surface area of the Araucaria kennellii whose soil around the roots was injected with the live bacteria solution of Masseillium E1-2h were significantly higher than those of the Araucaria kennellii whose soil around the roots was injected with the inactivated bacteria solution of Masseillium E1-2h. The results showed that Massilia E1-2h could promote the formation of Araucaria kennellii mycorrhizae and increase the number of Araucaria kennellii mycorrhizae, which might effectively promote its absorption of water and various mineral elements, increase the root substances of plants and improve the stress resistance of plants.
[0022] Table 1. Table of related indexes of Araucaria kennerii inoculated with Masseella E1-2h bacterial solution The above description is only a preferred embodiment of the present invention. All equivalent changes and modifications made according to the scope of the patent application of the present invention should fall within the scope of the present invention.
Claims
1. A strain of Massiliae that promotes the growth of Araucaria kennensis, characterized in that: The strain was named as Massillarisma ( Massilia sp. )E1-2h, was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on September 18, 2024, with the deposit number CGMCC No.31989, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.
2. A microbial agent for promoting the growth of Araucaria kennellii, characterized in that: Contains the Massillaria strain according to claim 1.
3. Use of the Massillaria strain according to claim 1 or the microbial agent according to claim 2 in promoting the growth of Araucaria kennellii.
4. The use according to claim 3, characterized in that: The method of promoting the growth of Araucaria kennellii includes promoting the formation of mycorrhiza of Araucaria kennellii, increasing the dry weight of the underground part of Araucaria kennellii, increasing the root surface area of Araucaria kennellii, and increasing the number of nodules per unit root surface area of Araucaria kennellii.
5. A method for promoting the growth of Araucaria kennellii, characterized in that: The Massillaria strain according to claim 1 is prepared into a bacterial liquid and injected into the soil around the root system of Araucaria kennellii.
6. The use according to claim 5, characterized in that: The method of promoting the growth of Araucaria kennellii includes promoting the formation of mycorrhiza of Araucaria kennellii, increasing the dry weight of the underground part of Araucaria kennellii, increasing the root surface area of Araucaria kennellii, and increasing the number of nodules per unit root surface area of Araucaria kennellii.