CN110862929B

CN110862929B - Gloeostereum incarnatum, microbial inoculum comprising same and application thereof

Info

Publication number: CN110862929B
Application number: CN201911249137.XA
Authority: CN
Inventors: 刘玮; 吴佳海; 刘姗; 张金莲; 赵小敏; 张嵚; 邓光华; 涂淑萍; 张露; 张微微; 胡松竹; 李响
Original assignee: Jiangxi Agricultural University
Current assignee: Jiangxi Agricultural University
Priority date: 2019-12-09
Filing date: 2019-12-09
Publication date: 2020-11-06
Anticipated expiration: 2039-12-09
Also published as: CN110862929A

Abstract

The invention provides a glomus affinis strain, a microbial inoculum comprising glomus affinis and application, and belongs to the technical field of microbial preparations. The preservation number of the glomus alatus Cc-01 is CGMCC No. 18593. The glomus alatus Cc-01 can form a mutual-benefiting symbiont with plants, so that the growth of the plant container seedlings is remarkably promoted, the nutritional status of the plant container seedlings is improved, and the capacity of the plant container seedlings for resisting external stress is improved. The application shows that the glomus alatus Cc-01 can remarkably promote the overground part growth of the plant container seedling, increase the leaf number, the plant height and the ground diameter of the plant container seedling, increase the total root length, the root surface area and the root tip number of the plant container seedling, and increase the root dry weight, the stem dry weight, the leaf dry weight and the total dry weight of the plant container seedling.

Description

Gloeostereum incarnatum, microbial inoculum comprising same and application thereof

Technical Field

The invention relates to the technical field of microbial preparations, in particular to a glomus affinis strain, a microbial inoculum comprising glomus affinis and application of the glomus affinis strain.

Background

Arbuscular Mycorrhizal Fungi (AMF) form a reciprocal symbiont with a plant nutrient root system to promote the absorption of nutrients such as host plant N, P, Cu and Zn, improve the permeability and water holding capacity of soil, enhance the drought resistance of plants, increase the accumulation of organic carbon in soil, improve the stress resistance of plants to stress such as water and nutrient deficiency, improve the growth of plants, improve the biomass of plants and the like. Sacculus closterium (Claroideis claroieum) is one of arbuscular mycorrhizal fungi, but the sacculus closterium which has the effect of promoting plant growth and is publicly reported in the prior art is less, and particularly, a sacculus closterium which has the effect of promoting plant growth is lacked.

Disclosure of Invention

The invention aims to provide glomus affinis, a microbial inoculum comprising glomus affinis and application thereof.

In order to achieve the above object, the present invention provides the following technical solutions:

the invention provides a strain of glomus alatus (Claroideis Clarioides) Cc-01 with the preservation number of CGMCC No. 18593.

The invention also provides a microbial inoculum comprising the glomus alatus.

Preferably, the effective spore number of the glomus affinis in the microbial inoculum is 10-80/g.

The invention also provides application of the glomus affinis or the microbial inoculum in the scheme in promoting growth of plant container seedlings, promoting plant photosynthesis and/or promoting plant transpiration, and the application comprises the following steps: filling matrix and microbial inoculum in the container from bottom to top in sequence, planting plant seedling, covering the matrix, watering, fixing seedling after the plant seedling grows stably, and culturing.

Preferably, the plant comprises euscaphis konishii hayata, cinnamomum kohlrabi or toona sinensis.

Preferably, the matrix comprises the following raw materials in parts by volume: 2.8-3.2 parts of garden soil, 0.8-1.2 parts of grass carbon and 0.8-1.2 parts of river sand.

Preferably, the container comprises a plastic flower pot.

Preferably, the number of seedlings per container is 1-2.

Preferably, the optical density of the culture is 750-850 mmol · m^-2·s^-1The illumination time is 15.5-16.5 h.d^-1The temperature is 25-27 ℃, and the humidity is 62-68%.

The invention has the beneficial effects that: the invention provides a strain of glomus alatus (Claroidei glomus muscularis) Cc-01 with the preservation number of CGMCC No. 18593. The glomus alatus Cc-01 can form a mutual-benefiting symbiont with plants, so that the growth of the plant container seedlings is remarkably promoted, the nutritional status of the plant container seedlings is improved, and the capacity of the plant container seedlings for resisting external stress is improved. Through application, the glomus pusillus Cc-01 can remarkably promote the overground part growth of plant container seedlings, increase the leaf number, the plant height and the ground diameter of the plant container seedlings, increase the total root length, the root surface area and the root tip number of the plant container seedlings, increase the root dry weight, the stem dry weight, the leaf dry weight and the total dry weight of the plant container seedlings, improve the absorption efficiency of plants on nutrients, assist the plants in absorbing the nutrients, promote the plant growth and promote the photosynthesis and the transpiration of the plants.

Biological preservation Instructions

The myxomycete (Claroideioglomus claroideum Cc-01) is preserved in the China general microbiological culture Collection center in 2019, 10 and 14 days, and the address is No. 3 of West Lu No.1 of Beijing Kogyo-sunward area, the institute of microbiology of China academy of sciences, and the preservation number is: CGMCC No. 18593.

Detailed Description

The invention provides a Claroideidoglomus claroideium Cc-01 strain with the preservation number of CGMCC No. 18593; the glomus affinis Cc-01 is separated from moso bamboo rhizosphere soil (the elevation is 740 meters) in the golden bamboo lawn area of the Lushan national-level natural protection area in Jiangxi province. In the invention, the glomus affinis and the host plant form a good symbiotic structure, and the growth of the host plant is promoted by promoting the nutrient absorption and improving the photosynthetic condition of the glomus affinis.

In the invention, the conidia of glomus basilicus has the following characteristics: the spore soil is singly or loosely gathered, spherical or nearly spherical, light yellow to yellowish brown, and 84.83-117.70 (average 87.20) mu m. Spore wall: 4 layers, L1 layer is colorless and transparent, thickness 0.50 ~ 2.00 μm, often fall off in mature spore and the surface is not uniform, sometimes only part of the residue is pink in Melzer's reagent dyeing reaction, becomes light blue in cotton blue reagent reaction; the thickness of the L2 layer is 1.20-2.60 mu m, the layer wall is light yellow to light yellow brown, the cotton blue reagent becomes light blue in the reaction, and the Melzer's reagent is not dyed in the reaction; the thickness of the L3 layer is 2.71-4.75 μm, the layer wall is light yellow to light yellow brown, the cotton blue reagent becomes light blue, and the Melzer's reagent is not dyed in the reaction; l4 is a layered wall with a thickness of 0.50-2.75 μm, single-layered or multi-layered, each layer having a thickness of about 0.50-1.00 μm, yellowish to yellowish-brown layered wall, blue in the cotton blue reagent reaction, light brown in the Melzer's reagent reaction. Hypha of Neurospora: the spore is cylindrical, is occasionally contracted, is transparent to light yellow, is 4.00-6.00 mu m wide, has three layers of walls, and falls off after the spores are mature; the L2 layers are transparent and light yellow and have the thickness of 1.50-3.00 mu m; the L3 layer is yellow brown, the thickness is 2.00-3.00 mu m, the connecting points are slightly thickened, and blocking connecting points are occasionally arranged.

Amplifying single spores of saccharum gloeosporium Cc-01 by adopting nested PCR (polymerase chain reaction), and performing first PCR amplification by adopting a fungus universal primer pair GeoA2-Geo 11; the second PCR amplification adopts the specific universal primer pair AML1-AML2 of the glomus proximal Cc-01. The 18S rDNA nucleotide sequence sequenced by the conidiobolus breve Cc-01 spore is shown as SEQ ID NO: 1, specifically: 5' -AAG CAC CGT TAC GTA TAG TCG CTC CAT TCG CGT ATC TTC AAG TTG TTG CAG TTAAAA AGC TCG TAG TTG AAT TTC GGG ATT GAC ACA TCG GTC GTG CCT TAA GGG GTA TGAACT GGT GTA GTC AAT TTC TCA CCT TCT GGA GAA CCG CGA TGC CCT TAA TTG GGT GTCACG GGG AAC CAG GAC CTT TAC TTT GAA AAA ATT AGA GTG TTT AAA GCA GGC ATT TTGCTT GAA TAC ATT AGC ATG GAA TAA TAA AAT AGG ACG GCA TGA TTC TAT TTT GTT GGTTTC TAG GAT CAC CGT AAT GAT TAA TAG GGA TAG TTG GGG GCA TTA GTA TTC AAT TGTCAG AGG TGA AAT TCT TGG ATT TAT TGA AGA CTA ACT ACT GCG AAA GCA TTT GCC AAGGAT GTT TTC ATT AAT CAA GAA CGA AAG TTA GGG GAT CGA AGA CGA TCA GAT ACC GTCGTA GTC TTA ACC ATA AAC TAT GCC GAC TAG GAA TCA GAC GAT GTT AAT TTT TTA ATGACT CGT TTG GCA CCT TAC GGG AAA CCA AAG TGT TTG GGT TCA AA-3'.

The invention also provides a microbial inoculum comprising the glomus alatus; the effective number of spores of glomus affinis in the microbial inoculum is preferably 10-80/g, more preferably 20-70/g, and most preferably 30-50/g.

In the invention, the microbial inoculum is preferably prepared by the following method: inoculating the glomus affinis in a matrix to obtain an expanding propagation matrix, planting corn and/or clover by using the expanding propagation matrix, fully propagating the glomus affinis between the expanding propagation matrix and a plant (corn and/or clover) root system, and finally harvesting a microbial inoculum consisting of dry soil of the expanding propagation matrix, hyphae outside the root, hyphae inside the root and an infected root section; the substrate comprises river sand and garden soil; the mass ratio of the river sand to the garden soil is preferably 1: 1; the substrate is preferably sterilized.

The invention also provides application of the glomus affinis or the microbial inoculum in the scheme in promoting growth of plant container seedlings, promoting plant photosynthesis and/or promoting plant transpiration, and the application comprises the following steps: filling a substrate and a microbial inoculum in a container from bottom to top in sequence, planting plant seedlings, then covering the substrate, watering, fixing seedlings after the plant seedlings grow stably, and culturing; the plant preferably comprises euscaphis konishii hayata, cinnamomum kohlrabi or toona sinensis.

In the invention, the application of the microbial inoculum in promoting the growth of the plant container seedlings preferably comprises promoting the overground part growth of the plant container seedlings and/or increasing the leaf number, the plant height and the ground diameter of the plant container seedlings and/or increasing the total root length, the root surface area and the root tip number of the plant container seedlings and/or increasing the dry weight, the stem weight, the leaf dry weight and the total dry weight of the plant container seedlings.

In the present invention, the matrix preferably comprises the following raw materials in parts by volume: 2.8-3.2 parts of garden soil, 0.8-1.2 parts of turf and 0.8-1.2 parts of river sand, and more preferably, the matrix comprises the following raw materials in parts by volume: 3 parts of garden soil, 1 part of grass peat and 1 part of river sand; the substrate is preferably screened through a 2mm screen; the pH value of the substrate is preferably 5.5-6.5, and more preferably 5.99; the organic matter content of the substrate is preferably 35-45 g-kg^-1More preferably 40 g/kg^-1(ii) a The organic carbon content of the substrate is preferably 65-75 g/kg^-1More preferably 68 g/kg^-1(ii) a The total phosphorus content of the substrate is preferably 0.5-0.9 g-kg^-1More preferably 0.8 g/kg^-1(ii) a The total nitrogen content of the substrate is preferably 1.7-2.1 g/kg^-1More preferably 1.9 g/kg^-1(ii) a The content of ammonium nitrogen in the matrix is preferably 13-18 mg/kg^-1More preferably 15 mg/kg^-1(ii) a The preferable content of nitrate nitrogen in the matrix is 4-4.5 mg/kg^-1More preferably 4.3 mg/kg^-1。

In the present invention, the matrix is preferably prepared by the following method: mixing the garden soil, the grass peat and the river sand, sieving the mixture by a 2mm sieve, and sterilizing the mixture to obtain a matrix; the mixing method is not particularly limited, and the uniform mixing is taken as the standard; the sterilization temperature is preferably 115-130 ℃, and more preferably 121 ℃; the pressure of the sterilization is preferably 0.1 MPa; the sterilization time is preferably 0.3-0.8 h, and more preferably 0.5 h; the sterilization mode is preferably high-temperature high-pressure steam sterilization.

In the present invention, the container preferably comprises a plastic flowerpot; the upper caliber, the lower caliber and the height of the plastic flowerpot meet the requirements of the planted plants.

In the invention, the container with the specification of upper caliber multiplied by lower caliber multiplied by high respectively being 9 multiplied by 6 multiplied by 8cm or the container with the specification of upper caliber multiplied by lower caliber multiplied by high respectively being 16 multiplied by 11 multiplied by 12.5cm (plastic flowerpot) is taken as a standard, the mass of the filling substrate of each container is preferably 60 to 300g, and more preferably 80 to 180 g; the mass of the filled microbial inoculum is preferably 50-70 g, and more preferably 60 g; the mass of the covering substrate is preferably 50-120 g, and more preferably 60-100 g; the number of the implanted plant seedlings is preferably 2-4, more preferably 3; the amount of water watered in each container is preferably 150-250 mL, and more preferably 200 mL; the number of seedlings per container is preferably 1-2. The method for obtaining the young plant is not particularly limited, and the conventional method in the field can be adopted.

In the present invention, the water holding capacity (mass ratio) of the soil to be cultured is preferably 18% to 25%, more preferably 20%; the optical density of the culture is preferably 750-850 mmol/m^-2·s^-1More preferably 800mmol · m^-2·s^-1(ii) a The illumination duration is preferably 15.5-16.5 h.d^-1More preferably 16 h.d^-1(ii) a The culture temperature is preferably 25-27 ℃, and more preferably 26 ℃; the humidity of the culture is preferably 62% to 68%, more preferably 65%.

The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.

Example 1 Hericium erinaceum

1. Selecting garden soil, peat and river sand according to the volume ratio of 3:1:1, sieving by a 2mm sieve, uniformly mixing, sterilizing by high-temperature (121 ℃) and high-pressure (0.1MPa) steam for 0.5h, and naturally cooling to obtain the matrix. The basic physicochemical properties of the matrix are as follows: pH value of 5.99 and organic matter content of 40.458 g/kg^-1(ii) a Organic carbon: 69.750 g/kg^-1(ii) a Total phosphorus: 0.754 g.kg^-1(ii) a Total nitrogen: 1.903 g/kg^-1(ii) a Ammonium nitrogen: 15.068 mg/kg^-1(ii) a Nitrate nitrogen: 4.209 mg/kg^-1。

2. Selecting the full Cinnamomum zeylanicum seeds collected in the same year, and adding 10% H₂O₂Soaking in the solution for 10min, sterilizing, and washing with sterile water₂O₂And (4) putting the residual liquid into a 4 ℃ thermostat, storing at low temperature for 1 month, taking out, embedding into sterile river sand sterilized at 121 ℃ for 30min, and culturing to obtain seedlings for later use.

3. The specification of the plastic flowerpot for the test is as follows: the upper caliber is multiplied by the lower caliber and multiplied by the height is respectively 9 multiplied by 6 multiplied by 8 cm; for a total of 20 pots. 180g of matrix is filled in each pot, then 60g of microbial inoculum (the microbial inoculum contains 600 saccharomyces subtenosus Cc-01 spores, Cc for short, the preservation number is CGMCC No.18593), 3 seedlings of the germinated cinnamomum longepaniculatum are planted, 60g of matrix is covered, 200mL of water is added to each pot, and 1 seedling is fixed after the growth of the seedlings is stable.

4. Quantitatively watering every week, keeping the water holding capacity of the soil at 20% (mass ratio), and keeping the optical density at 800 mmol.m^-2·s^-1The light is supplemented by an agricultural sodium lamp, the illumination time is 16 hours per day, the temperature is 26 ℃, and the humidity is 65%. Weighing once every three days, recording the growth conditions (plant height, ground diameter and leaf number) of the overground part of the cinnamomum kanahirai hance, observing the root development conditions (total root length, root surface area, root volume, average root diameter and root tip number) of the cinnamomum kanahirai hance, and counting the biomass (root dry weight, stem dry weight, leaf dry weight, total dry weight and root crown ratio) of the cinnamomum kanahirai hance. See tables 1, 2 and 3 for results.

Comparative example 1

The setup was the same as in example 1 except that no inoculum was inoculated. See tables 1, 2 and 3 for results.

Comparative example 2

The setup was the same as in example 1 except that the inoculum was replaced with a strain containing 600 mucosepalatus viscosus spores (Sv, accession number: CGMCC No.18594) isolated from Carcinia cottoninensis lawn Phyllostachys rhizosphere soil. See tables 1, 2 and 3 for results.

Comparative example 3

The setup was the same as in example 1 except that the inoculum was replaced with a mixture containing 300 mucosepalax spores Sv and 300 glomus proximal glomus spores Cc. See tables 1, 2 and 3 for results.

TABLE 1 Effect of inoculation with different bacterial agents on the growth of aerial parts of Cinnamomum zeylanicum

Note: data in the table are mean ± standard deviation. Different lower case letters in the same column represent significant differences at the 0.05 level and different upper case letters represent significant differences at the 0.01 level. The same applies below.

As can be seen from table 1, inoculation of AM fungi (glomus damascens and/or glomus mucosae) significantly promoted biomass growth of the cinnamomum longepaniculatum container seedlings, in which the number of leaves, plant height, and ground diameter of the container seedlings were significantly increased, as compared to the control (comparative example 1). The influence of the Sv-Cc mixed inoculation treatment on the height of the cinnamomum kanahirai hance is the largest, the height of the cinnamomum kanahirai hance is increased by 126% compared with a control, and the height of the cinnamomum kanahirai hance treated by Cc single inoculation treatment is increased by 97.8% compared with the control; cc single inoculation treatment has the best promotion effect on the ground diameter and leaf number of the cinnamomum longepaniculatum seedlings, increases 49.3% and 151.2% respectively compared with the control, and reaches a very significant level (p <0.01) statistically. Comprehensively judging, the growth promoting effect on the overground part of the monkeytree bark is best by the treatment of Sv-Cc mixed grafting and Cc single grafting.

TABLE 2 Effect of inoculating different bacterial agents on the development of the root System of Cinnamomum zeylanicum

As can be seen from Table 2, the AM fungi (glomus affinis and/or glomus mucosae) inoculation also has a relatively good promoting effect on the development of the root system of the cinnamomum longepaniculatum, the Cc single inoculation has the best promoting effect on the total root length, the root surface area and the number of root tips of the cinnamomum longepaniculatum seedlings, and compared with other treatments, the growth difference of the Cc single inoculation treated root tips of the cinnamomum longepaniculatum reaches a very significant level (p is less than 0.01); the Sv single grafting has a good promotion effect on two indexes, namely the root volume and the average root diameter of the cinnamomum kanahirai dunn seedlings, but the difference of the two indexes in comparison with the Cc single grafting treatment does not reach significance (p is more than 0.05).

TABLE 3 Effect of inoculating different bacterial agents on Cinnamomum zeylanicum biomass

As can be seen from Table 3, the inoculation of AM fungi (Gliocladium breve and/or Gliocladium viscidum) can significantly increase the biomass of different organs of the Cinnamomum zeylanicum container seedling. Cc Single graft treatment performed optimally (p >0.05) in root dry weight, shoot dry weight, leaf dry weight, and total dry weight.

Example 2 Toona sinensis

2. Selecting full euscaphis konishii hayata seeds collected in the same year, and adding 10% of H₂O₂Soaking in the solution for 10min, sterilizing, and washing with sterile water₂O₂Soaking the residual liquid in 65 deg.C boiled water, and continuously soaking in sterile water for 5 days (changing water every day) after natural cooling. Then putting the seeds into an incubator at 25 ℃ for accelerating germination.

3. The specification of the plastic flowerpot for the test is as follows: the upper caliber is multiplied by the lower caliber and multiplied by the height is respectively 9 multiplied by 6 multiplied by 8 cm; for a total of 20 pots. 120g of matrix is filled in each pot, then 60g of microbial inoculum is inoculated, (the microbial inoculum contains 600 saccharum nearmingming Cc-01 spores, Cc for short, and the preservation number is CGMCC No.18593), 3 euscaphis konishii seedlings with consistent growth vigor are respectively planted in each pot, 60g of matrix is covered, 200mL of water is added in each pot, and 1 seedling is fixed after the seedlings grow stably.

4. Quantitatively watering every week, keeping the water holding capacity of the soil at 20% (mass ratio), and keeping the optical density at 800 mmol.m^-2·s^-1The light is supplemented by an agricultural sodium lamp, the illumination time is 16 hours per day, the temperature is 26 ℃, and the humidity is 65%.

Comparative example 4

The setup was the same as in example 2 except that no inoculum was inoculated. The results of comparing example 2 with comparative example 4 are shown in tables 4 to 6.

Results of the experiment

1. Influence of inoculated saccaromyces juveniles Cc-01 on growth of euscaphis konishii hayata seedlings

As can be seen from table 4, the inoculation of saccharum nearculigii Cc-01 (Cc for short, example 2) can significantly promote the biomass growth of the euscaphis konishii hayata container seedlings compared with the control (comparative example 4), wherein the leaf number (increased by 219.2%), the plant height (increased by 215.6%) and the ground diameter (increased by 133.5%) of the container seedlings are significantly increased.

TABLE 4 influence of Pediobolus brevicaulis Cc-01 inoculation on overground growth of Euscaphis konishii

Note: the different letter representations in the same column differ significantly at the 0.05 level. (the same below)

As can be seen from Table 5, the inoculation of the target strain Gliocladium breve Cc can significantly improve the biomass of different organs of the euscaphis konishii container seedling. Proved that the inoculation of the gloriopsis cumingii Cc indeed improves the nutrient absorption efficiency of euscaphis konishii hayata. The root-cap ratio is an important index for representing the energy distribution of the plant, the lower the root-cap ratio is, the normal growth of the overground part of the plant can be maintained by using fewer root systems, and the higher the root-cap ratio is, the more the distribution of substances used for the root systems of the underground part of the plant to maintain the growth is proved. The root crown of the euscaphis konishii hayata after the inoculation of the target strain is 46.2 percent smaller than that of the euscaphis konishii hayata after the control treatment without inoculation, which proves that the euscaphis konishii hayata after the inoculation can maintain the biomass growth of the overground part of the euscaphis konishii hayata with less root system proportion, wherein microorganisms can play a role in assisting plants to absorb nutrients and promoting growth.

TABLE 5 Effect of Pediobolus brevicaulis Cc-01 inoculation on Toonae tonkinensis Biomass

2. Influence of inoculated sacculus nearing-Ming mildew Cc-01 on photosynthetic property of euscaphis konishii hayata seedlings

The net photosynthetic rate is the absorption of CO by plant photosynthesis₂Is subtracted from respiration to produce CO₂I.e. the actual photosynthetic rate of the plant. As can be seen from Table 6, inoculation of the target species significantly increased the net photosynthetic rate of the euscaphis konishii hayata container seedlings (by 158.1%, p)<0.05), promoting the photosynthesis to proceed.

The pores being CO₂The gas inlet and outlet channels of the blade, and the gas hole conductance Gs represents the CO entering the surface of the blade in unit time and per unit area₂The amount of (c) can reflect the degree of pore opening; intercellular CO₂Concentration (Ci) is CO in mesophyll intercellular spaces inside the leaves₂Concentration of external CO₂The balance of photosynthesis and respiration inside the leaf and various driving forces and resistances applied to the gas entering mesophyll cells; the transpiration rate Tr is the amount of water lost by the leaves through the transpiration in a certain unit time, and can reflect the strength of the transpiration of the plants. As can be seen from Table 6, after the target strain is inoculated, the stomatal conductance and the transpiration rate of euscaphis konishii hayata are both remarkably increased (respectively increased by 400.0 percent and p is increased by p<0.05；248.0％，p<0.05), intercellular CO₂The concentration also increased (6.35% increase).

TABLE 6 Effect of Pediobolus brevicaulis Cc-01 inoculation on photosynthetic Properties of Toona tonkinensis container seedlings

Example 3 Toona sinensis

1. Selecting garden soil, peat and river sand according to the volume ratio of 3:1:1, sieving by a 2mm sieve, uniformly mixing, sterilizing by high-temperature (121 ℃) and high-pressure (0.1MPa) steam for 0.5h, and naturally cooling to obtain the matrix. The basic physicochemical properties of the matrix are as follows: pH value of 5.02 and organic matter content of 32.47 g/kg^-1(ii) a Organic carbon: 55.98 g.kg^-1(ii) a Total phosphorus: 0.509g kg^-1(ii) a Total nitrogen: 1.25 g.kg^-1(ii) a Total potassium: 1.79 mg/kg^-1。

2. Selecting plump toona sinensis seeds collected in the same year, and diluting with 10% H₂O₂Soaking the solution for 5min, and removing residual H from the seeds with distilled water₂O₂The solution is washed and soaked in distilled water for one night for standby. Spreading the cooled sand in a tray, mixing with the soaked seeds, placing in an incubator for germination, keeping the temperature at 25 deg.C, keeping the humidity at 66%, and irradiating with sunlight for 12 hr, and irrigating the seeds with distilled water in the morning and evening every day. After 2 true leaves grow out, embedding the seeds in sterile river sand sterilized at 121 ℃ for 30min and cooled for culturing and seedling emergence preparationThe application is as follows.

3. The specification of the plastic flowerpot for the test is as follows: the upper caliber is 16 multiplied by 11 multiplied by 12.5cm, the lower caliber is multiplied by the height, and the water is used after being wiped and disinfected by 70 percent alcohol, and the total number of the water is 25 basins. 300g of substrate is loaded in each pot, then 60g of microbial inoculum is inoculated, (600 saccharomyces subtenosus Cc-01 spores, Cc for short, and the preservation number is CGMCC No.18593) is contained in the microbial inoculum), 3-4 toona sinensis seedlings with good growth vigor are respectively selected and transplanted in each pot, 120g of substrate is covered, and the seedlings are flattened and compacted. After the nursery stock grows stably, 1 plant is fixed.

Comparative example 5

The setup was the same as in example 3 except that no inoculum was inoculated. The results of comparing example 3 with comparative example 5 are shown in tables 7 to 9.

Results of the experiment

1. Influence of sacculus gloeosporus Cc-01 inoculation on growth of toona sinensis seedlings

As can be seen from table 7, inoculation of glomus Torulata (example 3) can promote toona pilea seedling biomass growth compared to control (comparative example 5), where the seedling ground diameter growth reaches significant levels (67.0%, p < 0.05). The plant height of the young toona sinensis seedlings is increased after the mucose sacculus isolariceus Sv-01 (increased by 13.3%).

TABLE 7 Effect of Gloeostereum nearing Cc-01 inoculation on overground growth of Toonae sinensis seedlings

Note: data in the table are mean ± standard deviation. The different lower case letters in the same column represent significant differences at the 0.05 level, as follows.

The inoculated saccharum minitans has a relatively good promoting effect on the development of the root system of the toona sinensis (table 8), the total root length of the toona sinensis seedlings after inoculation is increased by 15.1 percent (p is more than 0.05), the root surface area is increased by 81.9 percent (p is less than 0.05), the root volume is increased by 212.5 percent (p is less than 0.05), and the average root diameter is increased by 76.8 percent (p is less than 0.05).

TABLE 8 influence of Gliocladium breve Cc-01 inoculation on the development of the root System of Toona sinensis seedlings

As can be seen from table 9, inoculation of saccharum capsulatum Cc can significantly increase the biomass of different organs of the seedlings of toona sinensis: the dry weight of the root of the young Chinese toon seedlings is increased by 157.4 percent (p <0.05), the weight of the stem is increased by 138.9 percent (p <0.05), the dry weight of the leaves is increased by 32.1 percent (p >0.05), and the total dry weight is increased by 87.5 percent (p <0.05) after inoculation. The ratio of the root to the crown of the inoculated Chinese toon seedlings is increased by 73.5%, the difference is not obvious, the result shows that the sacculus nearing gming Cc-01 realizes the increase of the biomass of the Chinese toon through the balanced promotion of the ground and the underground parts, and the promotion effect of the sacculus neargming Cc-01 on the ground and the underground parts of the Chinese toon is higher than that of the ground and the overground parts.

TABLE 9 Effect of Gliocladium breve Cc-01 inoculation on Toonae sinensis seedling Biomass

The above examples show that the inoculation of the Tolypocladium gmelinii has a very good promoting effect on the growth of the container seedlings of the Cinnamomum zeylanicum, the euscaphis konishii or the toona sinensis.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Sequence listing

<110> university of agriculture in Jiangxi

<120> sacculus nearing open, microbial inoculum comprising same and application thereof

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<213> balloon mold (Claroideoglomus Clarioidum)

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gtcaatttct caccttctgg agaaccgcga tgcccttaat tgggtgtcac ggggaaccag 180

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atggaataat aaaataggac ggcatgattc tattttgttg gtttctagga tcaccgtaat 300

gattaatagg gatagttggg ggcattagta ttcaattgtc agaggtgaaa ttcttggatt 360

tattgaagac taactactgc gaaagcattt gccaaggatg ttttcattaa tcaagaacga 420

aagttagggg atcgaagacg atcagatacc gtcgtagtct taaccataaa ctatgccgac 480

taggaatcag acgatgttaa ttttttaatg actcgtttgg caccttacgg gaaaccaaag 540

tgtttgggtt caaa 554

Claims

1. The application of the glomus affinis or the microbial inoculum comprising the glomus affinis in promoting the growth of plant container seedlings, promoting plant photosynthesis and/or promoting plant transpiration comprises the following steps: filling a substrate and a microbial inoculum in a container from bottom to top in sequence, planting plant seedlings, then covering the substrate, watering, fixing seedlings after the plant seedlings grow stably, and culturing;

the glomus alatus is glomus alatus (Claroideis clarioides) Cc-01, and the preservation number is CGMCC No. 18593;

the effective spore number of the glomus affinis in the microbial inoculum is 10-80/g;

the plant is euscaphis konishii hayata, cinnamomum longepaniculatum or toona sinensis;

the matrix consists of the following raw materials in parts by volume: 2.8-3.2 parts of garden soil, 0.8-1.2 parts of grass carbon and 0.8-1.2 parts of river sand.

2. The use of claim 1, wherein the container comprises a plastic pot.

3. The use according to claim 2, wherein the number of seedlings per container is 1-2.

4. The use according to claim 1, wherein the optical density of the culture is 750 to 850 mmol-m^-2·s^-1The illumination time is 15.5-16.5 h.d^-1The temperature is 25-27 ℃, and the humidity is 62-68%.