CN114375640B - Method for promoting growth of camellia oleifera seedlings by using dark-color endophytic fungi - Google Patents
Method for promoting growth of camellia oleifera seedlings by using dark-color endophytic fungi Download PDFInfo
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Abstract
The invention discloses a method for promoting oil tea seedling growth by using dark-color alternate endophytic fungi, relates to the related field of oil tea seedling inoculation, and provides a new method for cultivating oil tea high-quality seedlings by using the effects of promoting host plant growth and improving host plant stress resistance after the dark-color alternate endophytic fungi and plants are subjected to co-culture. The method comprises the following steps: preparing DSE bacterial liquid; inoculating the colony of Acrocalymma vagum fungus to a sterilized potato glucose liquid culture medium on a sterile operation table, carrying out shake culture at 28 ℃ for 15d in a shaking table with the rotating speed of 120r/min, filtering and collecting mycelium pellets by using sterile gauze, washing the mycelium pellets by using sterile water for several times, and crushing the mycelium pellets to prepare a bacterial liquid with the concentration of 1 × 105 cfu/mL; step two: treating small seedlings or seeds of the camellia oleifera; step three: sterilizing the culture flowerpot and the culture medium; step four: soaking seeds; step five: carrying out pot culture; step six: and (5) seedling stage management and transplanting.
Description
Technical Field
The invention relates to the field related to inoculation of camellia oleifera seedlings, in particular to a method for promoting growth of camellia oleifera seedlings by using dark-color septate endophytic fungi.
Background
Dark septate endophytic fungi (DSE) are a group of endophytic fungi which are relatively concerned in recent years, and refer to a group of small soil fungi colonized in plant roots, and the fungi are generally characterized in that hyphae have obvious transverse septa and deep color, are widely present in root cortex, epidermis and vascular bundle tissue cells or intercellular spaces of healthy plants, can form microsclera in the plant cells or the intercellular spaces, and have no negative influence on the root tissues or the cells. A plurality of researches show that the DSE can promote the growth of plants, increase the tolerance of the plants to abiotic stress, and help the plants to resist drought, cold and heavy metal pollution. Acrocalyma vagum is one of DSEs, and the effects of promoting plant growth and increasing plant resistance are proved in astragalus, tobacco, holly, paper mulberry and red bean trees, so that the Acrocalyma vagum is a dark-color septate endophytic fungus which has a wide host range, an obvious growth promoting effect and capability of increasing plant resistance, and has a good development and utilization value.
The camellia oleifera is an important woody oil tree species in China, and has been cultivated and utilized for more than 2300 years in China. The oil tea has high economy. The camellia oil squeezed from the tea seeds contains more than 80% of unsaturated fatty acid, is very beneficial to human body and is called as 'east olive oil'; the tea seed cake is rich in tea polyphenol, tea saponin and tea polysaccharide, and can be developed into various washing products and organic feed and biological pesticide; the tea shell is also an industrial raw material for preparing active carbon and biological tannin extract. The oil tea industry has become one of the strong agricultural and industrial industries in China. The tea-oil tree seedlings grow slowly in the early period, fruits can be produced in 4-5 years, the tea-oil tree seedlings are not cold-resistant and cannot be planted in many high and cold areas, and the DSE is applied to cultivation of the tea-oil tree seedlings, so that the tea-oil tree seedlings have very important significance in promoting growth of the tea-oil tree seedlings, improving quality of the tea-oil tree seedlings, resisting adverse environments and the like. Therefore, the market urgently needs to develop a method for promoting the growth of the camellia oleifera seedlings by utilizing dark-colored separated endophytic fungi to help people to solve the existing problems.
Disclosure of Invention
The invention aims to provide a method for promoting the growth of camellia oleifera seedlings by using dark-color spaced endophytic fungi, which is mainly used for cultivating camellia oleifera seedlings by using the effects of promoting the growth of host plants and improving the stress resistance of the host plants after the dark-color spaced endophytic fungi and the plants are combined, so that a novel method is provided for cultivating high-quality camellia oleifera seedlings.
In order to achieve the purpose, the invention provides the following technical scheme: a method for promoting the growth of oil tea seedlings by using dark-color septate endophytic fungi comprises the following steps:
the method comprises the following steps: preparing DSE bacterial liquid; inoculating the colony of Acrocalymma vagum fungus to a sterilized potato glucose liquid culture medium on a sterile operation table, carrying out shake culture at 28 ℃ for 15d in a shaking table with the rotating speed of 120r/min, filtering and collecting mycelium pellets by using sterile gauze, washing the mycelium pellets by using sterile water for several times, and crushing the mycelium pellets to prepare a bacterial liquid with the concentration of 1 × 105 cfu/mL;
step two: treating small seedlings or seeds of the camellia oleifera;
step three: sterilizing the culture flowerpot and the culture medium;
step four: soaking seeds; selecting a container, washing with clear water, wiping the interior of the container with 75% alcohol, pouring the bacterial liquid into the container, placing the camellia oleifera seedlings or camellia oleifera seed seedlings with sterilized roots, and soaking for 1 hour;
step five: performing pot culture; putting the sterilized culture medium into a flowerpot, putting the camellia oleifera plantlets or camellia oleifera seeds soaked with the bacterium solution, covering the base parts of the plantlets or the camellia oleifera seeds with the sterilized culture medium, immediately irrigating the plantlets or the seeds with 50ml of the bacterium solution for 1 time, irrigating with sterile water, and putting the flowerpot into a greenhouse for culturing after the water is irrigated;
step six: seedling management and transplanting; and (3) putting the flowerpot into a greenhouse, culturing the seedlings according to the conventional method, supplementing water in time, supplementing 50ml of bacterial liquid for 1 time in the 2 nd week and the 3 rd week of culture, continuing conventional culture, and transplanting the seedlings in the mountain after 3 months.
Preferably, the treatment method of the camellia oleifera plantlets comprises the following steps: selecting camellia oleifera plantlets which are subjected to cuttage for about 1 year, firstly washing the roots of the plantlets in tap water to remove soil or matrix, then soaking the roots of the plantlets in 0.1% potassium permanganate solution for 10min, then washing with sterile water for 5 times, and then soaking in bacterial liquid.
Preferably, the treatment method of the camellia seed comprises the following steps: selecting a large container, soaking the camellia oleifera seeds in 0.1% potassium permanganate solution for 30min, cleaning the camellia oleifera seeds with sterile water for several times, spraying pure river sand with 0.1% potassium permanganate solution for wetting, paving river sand at the bottom of the large container to reach the height of 1/3 of the container, paving a layer of camellia oleifera seeds on the surface of the river sand, covering the river sand with a layer of river sand with the thickness of 5cm, repeating the operation until the container is filled, and covering the film on the large container for storage.
Preferably, the camellia seeds are harvested and used after three months, and the camellia seeds are in a radicle germination state.
Preferably, the Acrocalymma vagum is obtained by separating roots of a wild red bean tree, and the fungus has the effects of promoting plant growth, improving plant drought resistance, cold resistance and heavy metal pollution resistance.
Preferably, in the third step, the components and the proportion of the culture medium are yellow loam: vermiculite: perlite =3:1:1.
preferably, in the third step, the culture flowerpot is sterilized by wiping with 75% alcohol; the sterilization method of the culture substrate selects one of high-temperature high-pressure sterilization for 2h and 12kGy irradiation sterilization under the conditions of 121 ℃ and 0.1MPa, and ensures that the dark spaced endophytic fungi inoculation environment is carried out in the sterile substrate.
Preferably, in the fifth step, the sterilized culture medium is placed at a height of 2/3 of the height in the flowerpot.
Compared with the prior art, the invention has the beneficial effects that:
1. in the invention, because the number and the types of the microorganisms at the rhizosphere of the plant are more, space competition relations exist among the microorganisms, and the deep color spaced endophytic fungi can only exert the effect after forming symbiotic relations with the plant; therefore, the plants are planted in the field after being impregnated with the dark spaced endophytic fungi, so that the method has an active effect on the dark spaced endophytic fungi to occupy ecological sites and infection sites of other microorganisms in the soil, and can ensure that the dark spaced endophytic fungi have higher infection rate and form a good symbiotic relationship with the plants.
2. According to the final test result, the infection rate of the oil tea inoculated by the Acrocalymma vagum is between 66.67 and 76.67, and the infection rate of seed inoculation and inoculation is slightly higher than that of plantlet inoculation; the dry weight of the oil tea seedlings inoculated with the Acrocalymma vagum is always higher than that of the oil tea seedlings not inoculated with the Acrocalymma vagum regardless of the plant height of the oil tea seedlings under normal watering or water stress, and the growth of the small oil tea seedlings is promoted by inoculating the Acrocalymma vagum. In addition, the proline and soluble sugar content of the oil tea seedlings inoculated with the Acrocalymma vagum under normal watering and water stress conditions is higher than that of the oil tea seedlings not inoculated with the Acrocalymma vagum, and the malondialdehyde content of the oil tea seedlings not inoculated with the Acrocalymma vagum is lower than that of the oil tea seedlings not inoculated with the Acrocalymma vagum, so that the inoculated with the Acrocalymma vagum promotes the resistance of the oil tea seedlings to drought.
3. In the invention, according to the final test result, the inoculated Acrocalymma bigelovii tea-oil tree plantlet is higher than the non-inoculated Acrocalymma bigelovii tea-oil tree plantlet in plant height and plant dry weight, which indicates that the inoculated Acrocalymma bigelovii tea-oil tree plantlet can still play the effect of promoting the growth of the DSE under the field planting condition. In addition, the field cold tolerance index, the indoor cold tolerance index and the average cold tolerance index of the small oil tea seedlings inoculated with the Acrocalymma vagum are lower than those of the small oil tea seedlings not inoculated with the Acrocalymma vagum, and the cold tolerance of the small oil tea seedlings not inoculated with the Acrocalymma vagum is stronger than that of the small oil tea seedlings not inoculated with the Acrocalymma vagum. The method shows that under the condition of field planting, the inoculated Acrocalymma vagum oil tea seedling not only exerts the growth effect of the oil tea seedling, but also improves the cold resistance of the oil tea seedling and increases the stress resistance of the oil tea seedling to adverse environments.
Drawings
FIG. 1 is a diagram of root infection Acrocalymma vagum of the camellia oleifera plantlets of the invention;
FIG. 2 is a base sequence diagram of Acrocalymma vagum of the present invention.
Detailed Description
The present embodiment is implemented on the premise of the technical solution of the present invention, and a detailed implementation manner and a specific operation process are given, but the protection scope of the present invention is not limited to the following embodiments. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified.
Method for promoting growth of camellia oleifera seedlings by utilizing dark fungus with inner growth
1.1 The cuttage young seedling of Camellia oleifera is soaked with dark-color endophytic fungi
(1) Preparation of DSE bacterial liquid
A colony of Acrocalymma vagum fungi separated from the rhizosphere of a wild red bean tree is inoculated into a sterilized Potato Dextrose (PDB) liquid culture medium on a sterile operating platform, and is subjected to shaking culture at 28 ℃ in a shaking table at the rotating speed of 120r/min for 15d, so that the Acrocalymma vagum fungi are propagated in a large quantity.
Filtering with sterile gauze to collect mycelium pellet, washing with sterile water for several times, shaking off the mycelium pellet in shaking table, and crushing to obtain bacterial liquid with concentration of 1 × 105 cfu/mL.
(2) Treatment of camellia oleifera plantlets
Selecting camellia oleifera seedlings which are subjected to cuttage for about 1 year, firstly washing the roots of the seedlings to remove soil or matrix in tap water, soaking the roots of the seedlings in 0.1% potassium permanganate solution for 10min, then washing with sterile water for 5 times, and then soaking in bacterial liquid.
(3) Sterilization of cultured flowerpots and culture medium
The flowerpot is wiped with 75% alcohol and then dried.
The culture medium (yellow loam: vermiculite: perlite 3: 1).
(4) Seed soaking
Selecting a container, cleaning with clear water, and wiping the interior of the container with 75% alcohol.
Pouring the prepared bacterial liquid into a container, and putting the camellia oleifera plantlets with sterilized roots into the container, wherein the soaking time is 1 hour.
(5) Potted plant culture
Putting the sterilized matrix into a flowerpot with the height of about 2/3 of that of the flowerpot, putting 1-2 camellia oleifera seedlings soaked with the bacterial liquid according to the size of the flowerpot, covering the base parts of the seedlings with the sterilized matrix, immediately watering the seedlings with 50ml of the bacterial liquid for 1 time, watering with sterile water, and putting the flowerpot into a greenhouse for culturing after watering.
(6) Management and transplantation in seedling stage
And (4) putting the flowerpot into a greenhouse, culturing the seedlings according to the conventional method, and supplementing water in time.
And (3) additionally watering 50ml of bacterial liquid for 1 time in each flowerpot in the 2 nd week and the 3 rd week of culture, namely irrigating the bacterial liquid for 3 times totally, then continuing to culture the camellia oleifera seedlings in a conventional manner, and transplanting the camellia oleifera seedlings in the mountain after 3 months.
Sand-stored camellia oleifera seed is impregnated with dark-color septate endophytic fungi
(1) Preparation of DSE bacterial liquid
Will be separated from the root of wild red beanComing fromAcrocalymma vagumInoculating fungus colony to sterilized potato glucose (PDB) liquid culture medium on sterile operation table, shake culturing at 28 deg.C for 15d in shaking table at rotation speed of 120r/min,Acrocalymma vagumthe fungi are propagated in large quantities.
Filtering with sterile gauze to collect mycelium pellet, washing with sterile water for several times, shaking off the mycelium pellet in shaking table, and crushing to obtain bacterial liquid with concentration of 1 × 105 cfu/mL.
(2) Oil tea seed treatment
Selecting a large container, soaking the camellia oleifera seeds in 0.1% potassium permanganate solution for 30min, cleaning the camellia oleifera seeds with sterile water for several times, spraying pure river sand with 0.1% potassium permanganate solution for wetting, paving river sand at the bottom of the container to the height of 1/3 of the container, paving a layer of camellia oleifera seeds on the surface of the river sand, covering the camellia oleifera seeds with a layer of river sand (the thickness of 5 cm), repeating the operation until the container is filled, and covering the container with a film for storage.
During which time care is taken to maintain the temperature and humidity of the river sand and seeds. The seeds were harvested three months later, at which point the seeds had germinated radicles and some had germinated embryos.
(3) Sterilization of cultivation pots and culture media
The flowerpot is wiped with 75% alcohol and then dried.
The culture medium (yellow loam: vermiculite: perlite 3: 1).
(4) Seed soaking
Selecting a container, cleaning with clear water, and wiping the interior of the container with 75% alcohol.
Pouring the prepared bacterial liquid into a container, putting the seed seedlings stored in sand, and soaking for 1 hour.
(5) Potted plant culture
Putting the sterilized matrix into a flowerpot with the height of about 2/3 of that of the flowerpot, putting 1-3 seeds soaked with the bacterial liquid according to the size of the flowerpot, covering the seeds with the sterilized matrix, immediately irrigating the seeds for 1 time with 50ml of the bacterial liquid, and putting the flowerpot into a greenhouse for culturing after the sterile water is sufficiently irrigated.
(6) Management and transplantation in seedling stage
And (4) putting the flowerpot into a greenhouse, culturing the nursery stock according to the conventional method, and supplementing water in time.
And (4) additionally pouring 50ml of bacterial liquid into each flowerpot for 1 time in the 2 nd week and the 3 rd week of culture, then continuing conventional culture for 3 months, and selecting proper time to transplant the flowerpot to the mountain.
Assays to evaluate vaccination efficacy and to detect DSE efficacy
2.1 Growth-promoting drought-resisting test for camellia oleifera seedlings
The test site is the institute of biological research in Guizhou province, and the test material is Weining short-column oil tea.
The experiment set up 8 groups of treatments:
a1: inoculating Acrocalymma vagum oil tea seeds with normal water;
a2: normal water plus no inoculation of Acrocalymma vagum oil tea seeds;
a3: water stress + inoculation of Acrocalymma vagum oil tea seeds;
a4: water stress + no inoculation of Acrocalymma vagum Camellia seed;
a5: inoculating Acrocalymma vagum oil tea plantlets with normal water;
a6: normal water plus no inoculation of Acrocalymma vagum oil tea plantlets;
a7: water stress and inoculation of Acrocalymma vagum oil tea plantlets;
a8: water stress + no inoculation of Acrocalymma vagum Camellia oleifera plantlets.
The seedlings are 1 year old camellia seedlings, and the average height of the seedlings is 10.5cm. Normal water content means that the soil water content is 90%, water stress means that the soil water content is 50%, and the soil water content is controlled by adopting a weighing method. 100 seeds or 100 seedlings are inoculated in each treatment, 10 seedlings are randomly selected after 3 months of greenhouse culture, and the indexes of the root system DSE infection rate, growth index (plant height, plant dry weight), drought resistance index (proline and malondialdehyde) and the like are measured and repeated for 3 times. The detailed data are shown in Table 1.
TABLE 1 Effect of Acrocalymma vagum inoculation on Camellia oleifera seedling growth and drought resistance
Test results show that the infection rate of the oil tea inoculated by the Acrocalymma vagum is between 66.67 and 76.67, and the infection rate of seed inoculation and inoculation is slightly higher than that of seedlings. The dry weight of the oil tea seedlings inoculated with the Acrocalymma vagum is always higher than that of the oil tea seedlings not inoculated with the Acrocalymma vagum regardless of the plant height of the oil tea seedlings under normal watering or water stress, which shows that the inoculated Acrocalymma vagum promotes the growth of the small oil tea seedlings. In addition, the proline and soluble sugar content of the oil tea seedlings inoculated with the Acrocalymma vagum under the conditions of normal watering and water stress is higher than that of the oil tea seedlings not inoculated with the Acrocalymma vagum, and the malondialdehyde content is always lower than that of the oil tea seedlings not inoculated with the Acrocalymma vagum, which shows that the inoculation of the Acrocalymma vagum promotes the oil tea seedlings to improve the drought resistance.
Growth-promoting cold-resistance test for camellia oleifera seedlings
The cold resistance of the camellia oleifera inoculated strain seedlings is jointly identified by adopting two methods of field direct identification and indoor low-temperature treatment. The field test site is Weining furnace mountain town Xinfengcun, the indoor test site is Guizhou province biological research institute, and the test material is common oil tea.
The indoor low temperature treatment test was performed in 3 months. The experiment set up 2 treatments, B1: inoculating Acrocalymma vagum oil tea plantlets; b2: the seedlings of Acrocalymma vagum oil tea were not inoculated.
Randomly selecting 5 plantlets for pretreatment for 24h at 4 ℃ and then freezing for 5 h at-9 ℃ after culturing 30 plantlets in each treatment; after the freezing treatment, the seedlings are unfrozen and cultured for 3 h under the dark condition at the temperature of 4 ℃, and then cultured for 10d at room temperature, the freezing injury symptoms are observed, the freezing injury index CI (accurate to integral number) is calculated, and the cold resistance of the seedlings is graded, and the grading standard is shown in table 2.
TABLE 2 grading Standard of Cold resistance of Camellia oleifera
| Score of | Degree of freezing injury | Cold resistance |
| 3 | Freezing injury index less than or equal to 10 | Strong strength (S) |
| 4 | 10. Less than or equal to 20 | Is stronger |
| 5 | 20. The freeze injury index is less than or equal to 50 | In |
| 7 | Freezing injury index > 50 | Weak (weak) |
The field direct identification test is carried out in 9 months, namely 2 treated seedlings are transplanted to a Xinfeng village in a furnace mountain town of Weining county, and the growth and freezing damage conditions of 10 tea trees are investigated in 3 months next year. When red withered leaves or green withered leaves above 1/3 of the leaves on the middle and upper parts are damaged, the damaged conditions are graded according to the percentage of the number of the frozen leaves in the total number of the leaves to be investigated, and the grading standard is shown in a table 3.
TABLE 3 grading Standard of the oil tea Tree in the field freezing injury condition
| Rank of | Freezing injury condition |
| 0 | The area of the frozen leaves is less than or equal to 5 percent |
| 1 | The area of the frozen leaves is more than 5 percent and less than or equal to 15 percent |
| 2 | The area of the frozen leaves is more than 15 percent and less than or equal to 25 percent |
| 3 | The area of the frozen leaves is more than 25 percent and less than or equal to 50 percent |
| 4 | The area of the frozen leaves is more than 50 percent |
The weighting factors of field identification and indoor low-temperature treatment are respectively 0.5. The formula is as follows:
in the formula, n i The number of frozen plants at each stage, x i The number of freezing injury grades at each stage is N, the number of total investigated plants is N, and 4 is the highest damage grade.
The cold tolerance of each treated camellia oleifera seedling is identified according to the freezing injury index CI, and detailed data are shown in Table 4.
TABLE 4 evaluation of Cold tolerance of camellia oleifera seedlings treated differently
| Treatment of | Plant height (cm) | Plant dry weight (g) | Field CI | Indoor CI | Mean CI | Cold resistance |
| B1 | 12.5 | 0.84 | 25 | 25 | 25 | In |
| B2 | 8.8 | 0.67 | 55 | 60 | 57.5 | Weak (weak) |
The results in Table 4 show that the seedlings inoculated with the Acrocalymma vagum oil tea have higher values than those of the seedlings inoculated with the non-inoculated Acrocalymma vagum oil tea in terms of plant height and plant dry weight, and the inoculated Acrocalymma vagum oil tea seedlings can still play the effect of promoting the growth of the seedlings by the DSE under the field planting condition. In addition, the results also show that the value of the inoculated Acrocalymma vagum oil tea seedling in both field CI (cold resistance index) and indoor CI and average CI is lower than that of the unopened Acrocalymma vagum oil tea seedling, and the cold resistance of the inoculated Acrocalymma vagum oil tea seedling is stronger than that of the unoiled Acrocalymma vagum oil tea seedling, which indicates that under the condition of field planting, the inoculated Acrocalymma vagum oil tea seedling not only exerts the growth effect of the seedling, but also improves the cold resistance of the oil tea seedling, and increases the adverse environment resistance of the oil tea seedling.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Claims (8)
1. A method for promoting the growth of oil tea seedlings by using dark-color septate endophytic fungi is characterized by comprising the following steps:
the method comprises the following steps: preparing DSE bacterial liquid; inoculating the colony of Acrocalymma vagum fungus to a sterilized potato glucose liquid culture medium on a sterile operation table, carrying out shake culture at 28 ℃ for 15d in a shaking table with the rotating speed of 120r/min, filtering and collecting mycelium pellets by using sterile gauze, washing the mycelium pellets by using sterile water for several times, and crushing the mycelium pellets to prepare a bacterial liquid with the concentration of 1 × 105 cfu/mL;
step two: treating small seedlings or seeds of the camellia oleifera;
step three: sterilizing the culture flowerpot and the culture medium;
step four: soaking seeds; selecting a container, washing with clear water, wiping the interior of the container with 75% alcohol, pouring the bacterial liquid into the container, placing the camellia oleifera seedlings or camellia oleifera seed seedlings with sterilized roots, and soaking for 1 hour;
step five: performing pot culture; putting a sterilized culture medium into a flowerpot, putting the camellia oleifera plantlet or camellia oleifera seed soaked with the bacterium solution, covering the base of the plantlet or the camellia oleifera seed with the sterilized culture medium, immediately watering the plantlet or seed with 50ml of the bacterium solution for 1 time, watering with sterile water, and putting the flowerpot into a greenhouse for culturing after watering;
step six: seedling management and transplanting; and (3) putting the flowerpot into a greenhouse, culturing the nursery stock according to the conventional method, supplementing water in time, supplementing 50ml of bacterial liquid for 1 time in the 2 nd week and the 3 rd week of culture, continuing conventional culture, and transplanting the nursery stock in the mountain after 3 months.
2. The method for promoting the growth of camellia oleifera seedlings by using dark septate endophytic fungi according to claim 1, wherein the treatment method of the camellia oleifera seedlings is as follows: selecting camellia oleifera plantlets which are subjected to cuttage for about 1 year, firstly washing the roots of the plantlets in tap water to remove soil or matrix, then soaking the roots of the plantlets in 0.1% potassium permanganate solution for 10min, then washing with sterile water for 5 times, and then soaking in bacterial liquid.
3. The method for promoting the growth of camellia oleifera seedlings by utilizing the dark color septate endophytic fungi according to claim 1, wherein the camellia oleifera seeds are treated by the following method: selecting a large container, soaking the camellia oleifera seeds in 0.1% potassium permanganate solution for 30min, cleaning the camellia oleifera seeds with sterile water for several times, spraying pure river sand with 0.1% potassium permanganate solution for wetting, paving river sand at the bottom of the large container to reach the height of 1/3 of the container, paving a layer of camellia oleifera seeds on the surface of the river sand, covering the river sand with a layer of river sand with the thickness of 5cm, repeating the operation until the container is filled, and covering the film on the large container for storage.
4. The method of promoting the growth of camellia oleifera seedlings by using the dark-colored septate endophytic fungus as set forth in claim 1, wherein: the camellia oleifera seeds are harvested and used after three months, and the camellia oleifera seeds are in a radicle germination state.
5. The method of claim 1 for promoting the growth of camellia oleifera seedlings by using dark septate endophytic fungi, wherein the method comprises the following steps: the Acrocalymma vagum is obtained by separating roots of wild red bean trees.
6. The method of claim 1 for promoting the growth of camellia oleifera seedlings by using dark septate endophytic fungi, wherein the method comprises the following steps: in the third step, the components and the proportion of the culture medium are yellow loam: vermiculite: perlite =3:1:1.
7. the method of claim 1 for promoting the growth of camellia oleifera seedlings by using dark septate endophytic fungi, wherein the method comprises the following steps: in the third step, the culture flowerpot is wiped with 75 percent alcohol for disinfection; the sterilization method of the culture medium selects one of high-temperature high-pressure sterilization for 2h and 12kGy irradiation sterilization under the conditions of 121 ℃ and 0.1 MPa.
8. The method of promoting the growth of camellia oleifera seedlings by using the dark-colored septate endophytic fungus as set forth in claim 1, wherein: in the fifth step, the placing height of the sterilization culture medium is 2/3 of the height in the flowerpot.
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