Tobacco endophytic fungi capable of remarkably promoting growth of tobacco seedlings and application of tobacco endophytic fungi

Abstract

The invention discloses tobacco endophytic fungi capable of remarkably promoting growth of tobacco seedlings and application of the tobacco endophytic fungi. The preservation number of the strain is CGMCC (China General Microbiological Culture Collection Center) Tobacco is a leaf commercial crop, but in recent years, tobacco planting and production are often limited by multiple factors such as diseases and insect pests, heavy metal stress and drought stress, tobacco plants grow slowly in the seedling stage or the field stage, and the yield and quality of tobacco leaves are seriously threatened. The biomass and agronomic traits of the tobacco seedlings can be remarkably improved by applying the eight endophytic fungus liquid separated from the K326 to the soil of the overground parts and roots of the tobacco seedlings of different varieties in a watering manner.

Description

Technical Field

[0001] The invention relates to the field of tobacco endophytic fungi, and in particular to tobacco endophytic fungi capable of significantly promoting tobacco seedling growth and applications thereof. Background Art

[0002] Plant endophytes refer to microorganisms that live inside various tissues and organs or in the interstitial spaces of healthy plants at certain or all stages of their life history, generally including endophytic bacteria, endophytic fungi and endophytic actinomycetes.

[0003] At present, there are more than 120 species of endophytic bacteria isolated from plants, belonging to 54 genera, of which the most common are Pseudomonas, Bacillus, Enterobacter and Agrobacterium. In the study of endophytic actinomycetes, the most research is on Frankia, while the research on endophytic fungi is mostly on grasses, medicinal plants, marine plants, etc., but there are few studies on tobacco.

[0004] Endophytes live in tobacco for a long time, and the two have co-evolved. Studies have shown that tobacco endophytes can produce growth-promoting substances, help tobacco absorb mineral nutrients, and promote tobacco growth. Saito et al. found that Clostridium sp. contains a large amount of nitrogenase, which can play a nitrogen-fixing role in tobacco, provide inorganic nitrogen sources for tobacco, and thus improve the yield and quality of tobacco leaves. Yang Zhenfu et al. isolated 3 strains of Bacillus amyloliquefaciens from tobacco, which had a certain growth-promoting effect on tobacco seedlings. Jin Huiqing et al. showed that Fusarium strain YCEF005 and Acanthopanax strains YCEF053, YCEF191, YCEF193, and YCEF199 can improve the biomass and physiological indicators of tobacco plants. Therefore, tobacco endophytic fungi are a new way to improve tobacco leaf disease resistance, stress resistance, and improve tobacco leaf yield and quality.

[0005] In our study on the endophytic fungal flora of Yunnan tobacco dominant varieties K326 and Yunyan 87. A total of 154 endophytic fungi were isolated from tobacco K326 under different culture conditions, and identified as 40 genera and 72 species. Among them, 106 endophytic fungi were isolated from tobacco K326 under field growth conditions, identified as 37 genera and 62 species, 23 endophytic fungi were isolated from K326 cultured in water, identified as 8 genera and 11 species, and 25 endophytic fungi were isolated from K326 cultured in sterile soil, identified as 4 genera and 7 species. After screening, a total of 8 endophytic fungi significantly promoted the biomass and trait indicators of tobacco seedlings of different tobacco varieties. Summary of the invention

[0006] In order to solve the problems existing in the above-mentioned prior art, the object of the present invention is to provide tobacco endophytic fungi having the ability to significantly promote the growth of tobacco seedlings and applications thereof.

[0007] To achieve the above object, the technical solution of the present invention is:

[0008] The endophytic fungus that promotes the growth of tobacco seedlings is Cladosporium tenuissimu, and the preservation number of this strain is: CGMCC N0.41396. It was deposited on July 5, 2024 in the General Microbiology Center of China Microorganism Culture Collection Administration, located at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0009] The separation of tobacco endophytic fungi comprises:

[0010] A. Materials: K326 plants harvested from the field were divided into six parts: upper leaves, middle leaves, lower leaves, tobacco stems, taproots, and fibrous roots;

[0011] B. Disinfection: Upper leaves, middle leaves, and lower leaves: soak in 75% alcohol for 30 seconds, soak in 0.1% mercuric chloride for 15 seconds, and rinse with sterile water for 3 times; Tobacco stems: wipe the epidermis with 75% alcohol, and keep the cortex and pith; Thick roots and fibrous roots: soak in 75% alcohol for 60 seconds, soak in 0.1% mercuric chloride for 20 seconds, and rinse with sterile water for 3 times. The sterile water after the last rinse is streaked on the plate as a control to check whether the sample is thoroughly disinfected;

[0012] C. Inoculation and separation: add sterile water to the tobacco leaf part for grinding, and evenly spread the grinding liquid and residue on the surface of the culture medium; cut the tobacco stem part into 5-8mm small pieces with an inoculation knife, and spread them on the surface of the culture medium; cut the thick root part into 3-5mm small sections with an inoculation knife, and spread the cut surface downward on the surface of the culture medium; add sterile water to the fibrous root part for grinding, and evenly spread the grinding liquid and residue on the surface of the culture medium, and culture all the materials in a constant temperature incubator at 28℃ for 5-7 days to observe the growth of the colony; due to the different reproduction rates of different fungi, the faster growing strains are separated and purified first, and the slower growing strains are separated and purified after they are stable;

[0013] Solid culture: Use potato dextrose agar medium PDA (46 g / L), add streptomycin (10 mg / L) to prevent bacterial growth, culture temperature 28 ° C, culture time 5 to 7 days;

[0014] The separated different types of endophytic fungi were inoculated into PDA culture medium for purification, and the purification was repeated 2 to 3 times until the colonies were single, and the endophytic fungus C-4 was obtained after screening;

[0015] Identification of the above endophytic fungi: PCR amplification of rDNA ITS of the strain was performed using universal primers ITS1 / ITS4 for fungi, and the ITS PCR amplification products were sequenced. The obtained sequences were searched by BLAST in the GenBank nuclear sequence database, and it was found that the ITS region of the test strain had the highest homology with the reported Cladosporium exigua. Combined with morphological identification, the species of the endophytic fungus was determined;

[0016] The tobacco endophytic fungus is extremely fine spore C-4, which is green on PDA medium, with short hairs on the surface and slightly white hairs on the edge, which is thicker; there are few hyphae, and the conidiophores are upright, slightly curved and lateral conidia are oblong; due to the few hyphae and thick surface, the solid culture medium can be dug out appropriately during the purification process for continuous purification until the colony is single and pollution-free; during screening, the addition of hyphae to the liquid culture medium needs to be carried out under sterile conditions, and the inoculation loop is used to scrape lightly, and the liquid culture medium is inserted into the liquid culture medium to ensure that a certain amount of hyphae enters the liquid culture medium. After one addition is completed, the inoculation loop should be placed under an alcohol blowtorch to fully burn and sterilize, and then cooled for the next inoculation. The fungus is isolated from the tissue of fresh plants, and there are many endophytic fungi and bacteria in plant tissues, and the types and abundance of endophytic fungi in different tissues are also different. This strain is from 108 mixed endophytic fungi.

[0017] The endophytic fungus and the method for preparing the application bacterial liquid of the endophytic fungus are as follows: the endophytic fungus according to claim 1 is inoculated into a liquid culture medium, cultured on a shaker, with a rotation speed of 220 rpm, a culture temperature of 30°C, a culture time of 48 to 72 hours, and set aside; the liquid culture medium: potato glucose broth, wherein potato extract powder 5 g / L, glucose 20 g / L, and streptomycin 10 mg / L.

[0018] The bacterial solution is used for pouring into the soil matrix of the aboveground part and root of different varieties of tobacco seedlings to promote the growth of tobacco seedlings; after being cultured for a certain period of time, the bacterial solution is counted by a hemocytometer, added into sterile culture medium and uniformly diluted to 20,000,000 / ml, and 5 ml is applied to each part.

[0019] The bacterial liquid is applied when the tobacco seedlings grow 5 to 6 true leaves, and the biomass and agronomic traits of the tobacco seedlings are detected two weeks later; the tobacco seedlings are in the rooting stage when they grow to 5 to 6 true leaves, and the tobacco seedlings in this stage have the ability to live independently, the main root begins to develop and become thicker, and the lateral roots grow in large numbers, and the nutrient absorption capacity of the root system can be improved with sufficient space; the leaf vein network has been formed, the conductive tissue has been improved, and the tobacco seedlings have a certain synthesis capacity and nutrient transportation function; the tobacco seedlings are in the rooting stage when they grow to 5 to 6 true leaves, and the tobacco seedlings in this stage have the ability to live independently, the main root begins to develop and become thicker, and the lateral roots grow in large numbers, and the nutrient absorption capacity of the root system can be improved with sufficient space; the leaf vein network has been formed, the conductive tissue has been improved, and the tobacco seedlings have a certain synthesis capacity and nutrient transportation function.

[0020] The biomass and agronomic traits to be detected include: fresh weight, root weight, aboveground fresh weight, plant height, maximum leaf length, maximum leaf width, and root length.

[0021] Compared with the prior art, the beneficial effects of the present invention are:

[0022] The fungus obtained by the present invention was inoculated into tobacco seedlings of four different varieties, namely Honghua Dajinyuan, Yunyan 87, K326 and Yunyan 97. According to the evaluation of seven indicators such as tobacco seedling biomass and agronomic traits, the effect of the endophytic fungus on the growth of tobacco seedlings was further confirmed, providing basic support for further application in production. The experiment showed that C-4 has a significant growth-promoting effect, especially the most significant effect on promoting root growth. Strong roots in the seedling stage are very beneficial to the later growth of tobacco plants. Root size is an important criterion for judging whether seedlings are qualified in production. The four tobacco varieties have different degrees of growth in both the above-ground and underground parts, which can shorten the growth time of tobacco seedlings and ensure that the tobacco seedlings meet the requirements of high-quality tobacco seedlings before transplanting. Compared with the control (sterile culture medium treatment), the plant height growth rates of the four tobacco varieties Yunyan 87, Yunyan 97, K326 and Honghua Dajinyuan inoculated with C-4 were 14.70%, 24.77%, 23.80% and 24.17%, respectively; the maximum leaf length growth rates were 17.80%, 19.23%, 24.70% and 13.20%, respectively; the maximum leaf width growth rates were 18.17%, 15.00%, 15.87% and 10.33%, respectively; the root length growth rates were 55.63%, 53.63%, 14.43% and 10.33%, respectively; the underground fresh weight growth rates were 156.80%, 19.07%, 56.07% and 100.70%, respectively; the aboveground fresh weight growth rates were 68.1%, 40.87%, 64.73% and 29.67%, respectively. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 Tobacco seedling trays and sowing methods;

[0024] Figure 2 Morphology of endophytic fungi of 1 strain Yunyan 87;

[0025] Figure 3 1 Endophytic fungal fermentation diagram of Yunyan 87;

[0026] Figure 4 The effect of a tobacco endophytic fungus on the growth of different tobacco varieties;

[0027] Figure 5 Comparison of the growth of tobacco seedlings inoculated with endophytic fungi and those not inoculated with endophytic fungi. DETAILED DESCRIPTION

[0028] The technical solution of the present invention is further described in detail below in conjunction with the accompanying drawings and specific embodiments:

[0029] like Figure 1-5 As shown,

[0030] An endophytic fungus that promotes the growth of tobacco seedlings. The preservation number of this strain is: CGMCC N0.41396. It was deposited on July 5, 2024 at the General Microbiology Center of the China Microbiological Culture Collection Administration, located at No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

[0031] The separation of tobacco endophytic fungi comprises:

[0032] A. Materials: K326 plants harvested from the field were divided into six parts: upper leaves, middle leaves, lower leaves, tobacco stems, taproots, and fibrous roots;

[0033] B. Disinfection: Upper leaves, middle leaves, and lower leaves: soak in 75% alcohol for 30 seconds, soak in 0.1% mercuric chloride for 15 seconds, and rinse with sterile water for 3 times; Tobacco stems: wipe the epidermis with 75% alcohol, and keep the cortex and pith; Thick roots and fibrous roots: soak in 75% alcohol for 60 seconds, soak in 0.1% mercuric chloride for 20 seconds, and rinse with sterile water for 3 times. The sterile water after the last rinse is streaked on the plate as a control to check whether the sample is thoroughly disinfected;

[0034] C. Inoculation and separation: add sterile water to the tobacco leaf part for grinding, and evenly spread the grinding liquid and residue on the surface of the culture medium; cut the tobacco stem part into 5-8mm small pieces with an inoculation knife, and spread them on the surface of the culture medium; cut the thick root part into 3-5mm small sections with an inoculation knife, and spread the cut surface downward on the surface of the culture medium; add sterile water to the fibrous root part for grinding, and evenly spread the grinding liquid and residue on the surface of the culture medium, and culture all the materials in a constant temperature incubator at 28℃ for 5-7 days to observe the growth of the colony; due to the different reproduction rates of different fungi, the faster growing strains are separated and purified first, and the slower growing strains are separated and purified after they are stable;

[0035] Solid culture: Use potato dextrose agar medium PDA (46 g / L), add streptomycin (10 mg / L) to prevent bacterial growth, culture temperature 28 ° C, culture time 5 to 7 days;

[0036] The separated different types of endophytic fungi were inoculated into PDA culture medium for purification, and the purification was repeated 2 to 3 times until the colonies were single, and the endophytic fungus C-4 was obtained after screening;

[0037] Identification of the above endophytic fungi: PCR amplification of rDNA ITS of the strain was performed using universal primers ITS1 / ITS4 for fungi, and the ITS PCR amplification products were sequenced. The obtained sequences were searched by BLAST in the GenBank nuclear sequence database, and it was found that the ITS region of the test strain had the highest homology with the reported Cladosporium exigua. Combined with morphological identification, the species of the endophytic fungus was determined;

[0038] The tobacco endophytic fungus is extremely fine spore C-4, which is green on PDA medium, with short hairs on the surface and slightly white hairs on the edge, which is thicker; there are few hyphae, and the conidiophores are upright, slightly curved and lateral conidia are oblong; due to the few hyphae and thick surface, the solid culture medium can be dug out appropriately during the purification process for continuous purification until the colony is single and pollution-free; during screening, the addition of hyphae to the liquid culture medium needs to be carried out under sterile conditions, and the inoculation loop is used to scrape lightly, and the liquid culture medium is inserted into the liquid culture medium to ensure that a certain amount of hyphae enters the liquid culture medium. After one addition is completed, the inoculation loop should be placed under an alcohol blowtorch to fully burn and sterilize, and then cooled for the next inoculation. The fungus is isolated from the tissue of fresh plants, and there are many endophytic fungi and bacteria in plant tissues, and the types and abundance of endophytic fungi in different tissues are also different. This strain is from 108 mixed endophytic fungi.

[0039] The endophytic fungus and the method for preparing the application bacterial liquid of the endophytic fungus are as follows: the endophytic fungus according to claim 1 is inoculated into a liquid culture medium, cultured on a shaker, with a rotation speed of 220 rpm, a culture temperature of 30°C, a culture time of 48 to 72 hours, and set aside; the liquid culture medium: potato glucose broth, wherein potato extract powder 5 g / L, glucose 20 g / L, and streptomycin 10 mg / L.

[0040] The bacterial solution is used for pouring into the soil matrix of the aboveground part and root of different varieties of tobacco seedlings to promote the growth of tobacco seedlings; after being cultured for a certain period of time, the bacterial solution is counted by a hemocytometer, added into sterile culture medium and uniformly diluted to 20,000,000 / ml, and 5 ml is applied to each part.

[0041] The bacterial liquid is applied when the tobacco seedlings grow 5 to 6 true leaves, and the biomass and agronomic traits of the tobacco seedlings are detected two weeks later; the tobacco seedlings are in the rooting stage when they grow to 5 to 6 true leaves, and the tobacco seedlings in this stage have the ability to live independently, the main root begins to develop and become thicker, and the lateral roots grow in large numbers, and the nutrient absorption capacity of the root system can be improved with sufficient space; the leaf vein network has been formed, the conductive tissue has been improved, and the tobacco seedlings have a certain synthesis capacity and nutrient transportation function; the tobacco seedlings are in the rooting stage when they grow to 5 to 6 true leaves, and the tobacco seedlings in this stage have the ability to live independently, the main root begins to develop and become thicker, and the lateral roots grow in large numbers, and the nutrient absorption capacity of the root system can be improved with sufficient space; the leaf vein network has been formed, the conductive tissue has been improved, and the tobacco seedlings have a certain synthesis capacity and nutrient transportation function.

[0042] The biomass and agronomic traits to be detected include: fresh weight, root weight, aboveground fresh weight, plant height, maximum leaf length, maximum leaf width, and root length.

[0043] Example: Cultivation of plant material;

[0044] The invention selects four different varieties of tobacco coated seeds, namely Honghua Dajinyuan, Yunyan 87, K326 and Yunyan 97, and sows them on a 3*4 small seedling tray (such as Figure 2 ), 3 seeds were sown in each hole, the nutrient soil used for seedling cultivation was sterilized by high pressure, and the cells were cultured in an incubator under alternating light and darkness, with 16 hours of light cultivation, 18000LX of light intensity, and 28°C; and 8 hours of dark cultivation, at 16°C; nutrient solution was added to the bottom of the seedling tray, and the nutrient solution needed to be replenished every two days.

[0045] Application of endophytic fungi liquid irrigation

[0046] Take a fungus block of about 1.5 cm2 obtained by separation and purification and inoculate it into 100 ml liquid culture medium. The culture medium is placed in a 250 ml conical flask and cultured in a shaking incubator. The culture temperature is 30°C, the rotation speed is 220 rpm, and the culture time is 48 to 72 hours. When the tobacco seedlings grow to 5 to 6 true leaves, take the bacterial solution to water the aboveground part and root soil of the tobacco seedlings. Before watering, keep one tobacco seedling in each hole in the seedling tray to ensure that the growth of each tobacco seedling is consistent. Take 3 ml of bacterial solution from each hole and water the tobacco seedlings. Shake the bacterial solution evenly before taking it. After watering, cover the top of the seedling tray with a transparent isolation cover and continue to culture in an incubator. Set up a control tobacco seedling, culture the control tobacco seedling normally without watering the bacterial solution. When the tobacco seedlings grow to 7 true leaves, measure 6 indicators of the tobacco seedlings, including the fresh weight of the underground part, the fresh weight of the aboveground part, the plant height, the maximum leaf length, the maximum leaf width, and the root length.

[0047] Effects of Cladosporium tenuissimum C-4 on the Growth of Tobacco Seedlings of Different Varieties

[0048] The results showed that C-4 had a significant growth-promoting effect. Compared with the control, the plant height growth rates of the four tobacco varieties Yunyan 87, Yunyan 97, K326 and Honghua Dajinyuan inoculated with C-4 were 14.70%, 24.77%, 23.80% and 24.17%, respectively; the maximum leaf length growth rates were 17.80%, 19.23%, 24.70% and 13.20%, respectively; the maximum leaf width growth rates were 18.17%, 15.00%, 15.87% and 10.33%, respectively; the root length growth rates were 55.63%, 53.63%, 14.43% and 10.33%, respectively; the underground fresh weight growth rates were 156.80%, 19.07%, 56.07% and 100.70%, respectively; the aboveground fresh weight growth rates were 68.1%, 40.87%, 64.73% and 29.67%, respectively.

[0049] The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that are not conceived through creative work should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope defined in the claims.

Claims

1. An endophytic fungus for promoting tobacco seedling growth, characterized in that: The deposit number of the strain is: CGMCC No. 41396; the separation of the tobacco endophytic fungi includes: A. Materials: K326 plants harvested from the field were divided into six parts: upper leaves, middle leaves, lower leaves, tobacco stems, taproots, and fibrous roots; B. Disinfection: Upper leaves, middle leaves, and lower leaves: soak in 75% alcohol for 30 seconds, soak in 0.1% mercuric chloride for 15 seconds, and rinse with sterile water for 3 times; Tobacco stems: wipe the epidermis with 75% alcohol, and keep the cortex and pith; Thick roots and fibrous roots: soak in 75% alcohol for 60 seconds, soak in 0.1% mercuric chloride for 20 seconds, and rinse with sterile water for 3 times. The sterile water after the last rinse is streaked on the plate as a control to check whether the sample is thoroughly disinfected; C. Inoculation and separation: add sterile water to the tobacco leaf part for grinding, and evenly spread the grinding liquid and residue on the surface of the culture medium; cut the tobacco stem part into 5-8mm small pieces with an inoculation knife, and spread them on the surface of the culture medium; cut the thick root part into 3-5mm small sections with an inoculation knife, and spread the cut surface downward on the surface of the culture medium; add sterile water to the fibrous root part for grinding, and evenly spread the grinding liquid and residue on the surface of the culture medium, and culture all the materials in a constant temperature incubator at 28℃ for 5-7 days to observe the growth of the colony; due to the different reproduction rates of different fungi, the faster growing strains are separated and purified first, and the slower growing strains are separated and purified after they are stable; Solid culture: Use potato dextrose agar medium PDA (46 g / L), add streptomycin (10 mg / L) to prevent bacterial growth, culture temperature 28 ° C, culture time 5 to 7 days; The separated different types of endophytic fungi were inoculated into PDA culture medium for purification, and the purification was repeated 2 to 3 times until the colonies were single, and the endophytic fungus C-4 was obtained after screening; Identification of the above endophytic fungi: PCR amplification of rDNA ITS of the strain was performed using universal primers ITS1 / ITS4 for fungi, and the ITS PCR amplification products were sequenced. The obtained sequences were searched by BLAST in the GenBank nuclear sequence database, and it was found that the ITS region of the test strain had the highest homology with the reported Cladosporium exigua. Combined with morphological identification, the species of the endophytic fungus was determined; The tobacco endophytic fungus is the extremely fine spore C-4, which is green on the PDA culture medium, with short hairs on the surface with slightly white hairs on the edges, and is relatively thick; the hyphae are few, the conidiophores are upright, and the slightly curved conidia are lateral and oblong; due to the small number of hyphae and the thick surface, part of the solid culture medium can be dug out appropriately during the purification process for continuous purification until the colony is single and pollution-free; during screening, the hyphae need to be added to the liquid culture medium under sterile conditions, and the inoculation loop is used to gently scrape and insert it into the liquid culture medium for immersion to ensure that a certain amount of hyphae enters the liquid culture medium. After completing one addition, the inoculation loop should be placed under an alcohol blowtorch for sufficient burning and sterilization, and then cooled for the next inoculation. The fungus is isolated from the tissues of fresh plants, and there are many endophytic fungi and bacteria in plant tissues, and the types and abundances of endophytic fungi in different tissues are also different. The strain is from 108 mixed endophytic fungi.

2. The endophytic fungus according to claim 1, characterized in that: The method for preparing the application bacterial liquid of the endophytic fungus is to inoculate the endophytic fungus described in claim 1 into a liquid culture medium, culture it on a shaker, at a rotation speed of 220 rpm, a culture temperature of 30° C., a culture time of 48 to 72 hours, and set it aside; the liquid culture medium: potato glucose broth, wherein 5 g / L potato extract powder, 20 g / L glucose, and 10 mg / L streptomycin.

3. The endophytic fungus according to claim 2, characterized in that The bacterial solution of endophytic fungi is used to water the aboveground part and root matrix soil of different varieties of tobacco seedlings to promote the growth of tobacco seedlings; after being cultured for a certain period of time, the bacterial solution is calculated by a hemocytometer, added to sterile culture medium and diluted to 20,000,000 / ml, and 5 ml is applied to each part.

4. The endophytic fungus according to claim 2, characterized in that The bacterial liquid of endophytic fungi was applied when the tobacco seedlings grew 5 to 6 true leaves, and the biomass and agronomic traits of the tobacco seedlings were tested two weeks later; the tobacco seedlings were in the rooting stage when they grew 5 to 6 true leaves, and the tobacco seedlings at this stage had the ability to live independently, the main root began to develop and become thicker, and lateral roots occurred in large numbers, and the nutrient absorption capacity of the root system could be improved with sufficient space; the leaf vein network had been formed, the conductive tissue had been perfected, and it had a certain synthetic capacity and nutrient transport function; the tobacco seedlings were in the rooting stage when they grew 5 to 6 true leaves, and the tobacco seedlings at this stage had the ability to live independently, the main root began to develop and become thicker, and lateral roots occurred in large numbers, and the nutrient absorption capacity of the root system could be improved with sufficient space; the leaf vein network had been formed, the conductive tissue had been perfected, and it had a certain synthetic capacity and nutrient transport function.

5. The endophytic fungus according to claim 4, characterized in that The biomass and agronomic traits to be detected include: fresh weight, root weight, aboveground fresh weight, plant height, maximum leaf length, maximum leaf width, and root length.

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