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Artificial cultivating method for advantageous symbiotic mycorrhizal fungi of Castanea henryi (Skam) Rehd. et Wils.

A symbiotic mycorrhizal and artificial culture technology, applied in microorganism-based methods, biochemical equipment and methods, horticultural methods, etc., can solve the problems of difficulty in the isolation and cultivation of mycorrhizal fungi, stunted growth and development, and complex symbiotic relationships. Large diameter, high mycorrhizal infection rate, and the effect of improving physiological activity

Active Publication Date: 2015-11-25
CENTRAL SOUTH UNIVERSITY OF FORESTRY AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the conventional grafting of chestnut seedlings, due to the lack of mycorrhizal fungi that can co-exist with the root system of chestnut in the seedbed or medium, the conventional grafted seedlings transplanted to the field have low survival rate and slow growth
However, the symbiotic relationship between the root system of Chestnut chestnut and ectomycorrhizal fungi is very complicated, and the isolation and culture of many mycorrhizal fungi is difficult, and the mycorrhizal research of Chestnut chestnut has not been reported yet.

Method used

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  • Artificial cultivating method for advantageous symbiotic mycorrhizal fungi of Castanea henryi (Skam) Rehd. et Wils.
  • Artificial cultivating method for advantageous symbiotic mycorrhizal fungi of Castanea henryi (Skam) Rehd. et Wils.

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Experimental program
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Effect test

Embodiment 1

[0033] A method for artificially cultivating mycorrhizal fungus, comprising the following steps:

[0034] (1) Collection and tissue separation of fruiting bodies of wild mycorrhizal fungi:

[0035] Under the 30-year-old chestnut garden, collect fresh, strong, medium-mature wild mycorrhizal fruiting bodies and the accompanying soil and root system of the chestnut. After removing surface debris, put them into pots without squeezing. Within 48 hours After taking it back, soak it in 75% alcohol for 5 seconds for disinfection, and then wash it twice with sterile water; on the sterile operating table, cut the cleaned and disinfected fruiting body into two halves longitudinally, and take out the cap with tweezers At the intersection with the stipe, a mung bean-sized piece of bacterial flesh tissue is obtained;

[0036] (2) Mycelia cultivation and purification:

[0037] The modified MMN plate medium includes the following components: CaCl 2 0.045g, MgSO 4 0.15g, NaCl 0.025g, the c...

Embodiment 2

[0068] The present embodiment provides a kind of artificial culture method of mycorrhizal fungus, compared with embodiment 1, only there is following difference:

[0069] In step (2), the improved MMN plate culture medium comprises the following components: CaCl 2 0.06g, MgSO 4 0.16g, NaCl 0.025g, the concentration is 1% FeCl 3 1.2ml, KH 2 PO 4 0.6g, Vitamin B1100μg, KNO 3 0.25g, 100ml of 12Be' wort, 20g of mannitol, 0.20g of citric acid, 20g of agar, 900ml of distilled water;

[0070] In step (3), the difference between the composition of the liquid improved MMN medium and the solid improved MMN medium described in this embodiment is that it does not contain agar; inoculate 6 diameter 4mm bacterium cakes in every 100ml liquid improved MMN medium, Shake culture at 100r / min for 10 days at a constant temperature of 25°C to obtain bacterial liquid;

[0071] In step (4), the mass ratio of peat soil, corn flour, and perlite forming the expanded culture material is 7:1.5:1.5; ...

Embodiment 3

[0073] The present embodiment provides a kind of artificial culture method of mycorrhizal fungus, compared with embodiment 1, only there is following difference:

[0074] In step (2), the improved MMN plate culture medium comprises the following components: CaCl 2 0.04g, MgSO 4 0.12g, NaCl 0.025g, the concentration is 1% FeCl 3 1.2ml, KH 2 PO 4 0.4g, Vitamin B1100μg, KNO 3 0.25g, 100ml of 12Be' wort, 20g of mannitol, 0.20g of citric acid, 20g of agar, 900ml of distilled water;

[0075] In step (3), the difference between the composition of the liquid improved MMN medium and the solid improved MMN medium described in this embodiment is only that it does not contain agar; in every 100ml of liquid improved MMN medium, 4 bacterium cakes with a diameter of 4mm are inoculated, Under the constant temperature condition of 25°C, shake culture at 100r / min for 15 days to obtain the bacterial liquid;

[0076] In step (4), the mass ratio of peat soil, corn flour, and perlite forming ...

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Abstract

The invention relates to an artificial cultivating method for advantageous symbiotic mycorrhizal fungi of Castanea henryi (Skam) Rehd. et Wils. The method comprises the following steps: (1) taking a fruiting body of mycorrhizal fungi of Castanea henryi (Skam) Rehd. et Wils., and collecting fungus context issue blocks at the intersecting part of a fungus cap and a fungus stipe; (2) putting the fungus context tissue blocks in a culture plate, culturing the fungus context tissue blocks at the temperature of 25-28 DEG C until mycelia are germinated by the fungus context issue blocks, transferring the mycelia, and then cultivating the mycelia at the temperature of 25-28 DEG C until the mycelia grow to cover the culture plate to obtain purified mycelia; (3) selecting the purified mycelia, and inoculating the selected purified mycelia to an improved MMN liquid culture medium for fermental cultivation, thus obtaining a bacterium solution; (4) inoculating the bacterium solution to an enlarged cultivating material, and cultivating the purified mycelia at the temperature of 25-28 DEG C until the purified mycelia grow all over a cultivating container to obtain mycorrhizal fungi inoculant of Castanea henryi (Skam) Rehd. et Wils. The artificial cultivating method for advantageous symbiotic mycorrhizal fungi of Castanea henryi (Skam) Rehd. et Wils. is simple in operation; when mycorrhizal fungi cultivated and obtained through the artificial cultivating method is inoculated to seedlings of Castanea henryi Rehd. et Wils., the impregnating rate is high.

Description

technical field [0001] The invention belongs to the field of artificial cultivation of mycorrhizal fungi, and in particular relates to a method for artificial cultivation of mycorrhizal fungi. Background technique [0002] Ectomycorrhizalfungi (ECMF) is formed by the symbiosis of plant vegetative roots and soil fungi, and it is ubiquitous in nature. Ectomycorrhizal fungi can expand the area of ​​host plant roots to absorb nutrients, produce a variety of organic acids, enzymes, antibiotics and other substances, and accelerate the cycle of organic matter and inorganic matter such as nitrogen, phosphorus, potassium, calcium, and magnesium in the ecosystem. Improve the nutritional status of planting soil, promote the growth of host plants, and at the same time have an antagonistic effect on root soil-borne diseases, and can enhance the stress resistance of plants. Ectomycorrhizal fungi affect the composition and quantity of microorganisms in the rhizosphere of host plants. The ...

Claims

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Application Information

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IPC IPC(8): C12N1/14A01G7/06A01G1/00C12R1/645
Inventor 袁德义刘冬明邹锋朱周俊张旭辉谭露曼
Owner CENTRAL SOUTH UNIVERSITY OF FORESTRY AND TECHNOLOGY
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