CN1408213A - Method for preparing forest mycorrhizal fungi preparation from different area and raw material - Google Patents
Method for preparing forest mycorrhizal fungi preparation from different area and raw material Download PDFInfo
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- CN1408213A CN1408213A CN 01128174 CN01128174A CN1408213A CN 1408213 A CN1408213 A CN 1408213A CN 01128174 CN01128174 CN 01128174 CN 01128174 A CN01128174 A CN 01128174A CN 1408213 A CN1408213 A CN 1408213A
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Abstract
The present invention relates to the method of forest mycorrhizal fungi preparation from different area and raw material. Matrix saw dust, bran, pearlite, turf, etc. in certain proportion are mixed, the mixture is further mixed with inorganic salt, partial nutrient elements and water to compound nutritious liquid with water conent of 50-70%, the nutritious liquid is packed, pressurized steam sterilized and cooled, and mycorrhizal fungi thallus produced via two-stage biological fermentation and in amount of 2-10% is inoculated into the matrix and constant temperature cultured for 10-30 days. The preparation process of the present invention is not limited by the raw material source and has low cost.
Description
Technical field
The present invention relates to fungal inoculant, the preparation method of the forest mycorrhizal fungi microbial inoculum that specifically a kind of area is different with raw material sources.
Background technology
Mycorhiza is a kind of universal phenomenon of natural world fungi and plant symbiosis.Mycorhiza can promote the absorption of plant to moisture and nutriment, produces bioactivator, strengthens the resistance of plant, and under certain plants or particular environment condition, the existence of mycorhiza can become the decisive factor that influences growth and development of plants.Thereby mycorhiza is widely used in agricultural and forestry production as a kind of biotechnology.Mycorrhizal fungi vegetative mycelium with under the pure culture state carries out artificial infection, quickens the nursery stock mycorhiza and forms, and with the promotion seedling growth, thereby has improved seedling quality.Research mycorrhizal fungi nutritional condition and microbial inoculum production technology realize the artificial Mycorrhizal of forest seedling, and this has broad application prospects in breeding and afforestation, particularly in the afforestation, has irreplaceable effect.Therefore, cause domestic and international mycorhiza expert, scholar's very big concern, and introduce a fine variety, grow seedlings, aspect such as afforestation and environmental protection does a lot of work, and obtained remarkable result.But when on producing, using the mycorhiza technology, the preparation of microbial inoculum is finished in the laboratory often, the wheat bran of the main 60-70% of employing matrix weight replenishes as nitrogenous source as nitrogenous source or with offcuts rice chaff, carbon source is based on weed tree sawdust, and microbial inoculum need carry or check the actual field application, cause microbial inoculum cost costliness, make the promotion and application of the new technology of this practicality be subjected to serious restriction.
Summary of the invention
In order to overcome the above-mentioned end of suppressing, the purpose of this invention is to provide the preparation method that a kind of cost is low, be not subjected to the regional forest mycorrhizal fungi microbial inoculum of limitting, being suitable for applying.
To achieve these goals, technical scheme of the present invention is: by weight each component of mixed preparing, can prepare according to the following procedure:
1) the matrix collocation mixes: matrix comprises that 3-4 part wood chip and 1-2 part wheat bran mix;
2) nutrient solution preparation: by adding diammonium hydrogen phosphate 0.1-0.5g, glucose 2-10g, calcium chloride 0.01-0.08g, magnesium sulfate 0.01-0.2g, ferric trichloride 0.001-0.005g, sodium chloride 0.05-0.3g, Cobastab in the 1 liter water
10.01-0.05g be made into nutrient solution with potassium dihydrogen phosphate 0.5-5g and/or dipotassium hydrogen phosphate 0.32-3.2g;
3) sterilization: above-mentioned nutrient solution and matrix are puddled, and making the water content of substrate after puddling is 50-70%, and packing is sealed, and adds to depress at 105-125 ℃ and carries out steam sterilizing 1-2 hour, cools off standby;
4) thalline preparation: bacterial classification and medium after the one-level of learning from else's experience is cultivated, be 2-5 with bacterial classification with the mixed proportion of medium: 100 mix, and produce thalline with the two stage biological fermentation method;
5) fungicide preparation: the matrix after will sterilizing was inoculated with the biomass of its 2-10%, at 23-30 ℃ of following constant temperature culture 10-30 days.
Also can add the adding of 10-70% that accounts for the matrix gross weight in the described matrix and fill agent, add fill agent be in the peat composed of rotten mosses, wheat tangerine, straw, rice husk, the corn straw one or more; Also can add in the described matrix and account for matrix gross weight 10-50% perlite as carrier; Also can select to add one or more following materials in the described nutrient solution preparation: yeast extract 0.05-0.3g, peptone 1.0-3.0g, ferrous sulfate concentration is 10-50ppm, and zinc sulphate concentration is 10-50ppm, and sodium molybdate concentration is 10-50ppm;
Described wood chip is one or more mixing in birch bits, larch wood chip, maple bits, the masson pine wood chip;
Need before mixed substrate, to steam more than 30 minutes in advance when described wood chip is the needle wood chip or normal pressure steams more than 2 hours in advance, carry out gas simultaneously and discharge with 120 degrees centigrade of steam.
Described bacterial classification is the forest mycorrhizal fungi: one or more mixing in kakgeusia breast mushroom, the sliding rust of burr umbrella, ash ring Suillus bovines, pinecone beef liver, high handle mushroom, the reddish brown cortina bacterium.
The present invention has following advantage
1. material source is extensive, and cost is low.The material that the present invention prepares microbial inoculum is the dead meal that produces in the Wood Processing process---wood chip, offcuts of crops (as wheat tangerine, straw, rice husk, corn straw) and construction material perlite, the peat composed of rotten mosses marshy, the wheat bran of the main 60-70% of employing matrix weight replenishes as nitrogenous source as nitrogenous source or with offcuts rice chaff and compares in these materials and the prior art, does not need highly to provide funds and be easy to get.
2. the present invention's operation is not subjected to the restriction in area.Because the forest mycorrhizal fungi is the symbiotrophy characteristics, has saprophytic characteristic again, so the wood chip of trees is them the naturally first-selected carrier matrix that grows.But because, the difference in area, raw-material kind is also inequality, and the present invention adjusts the technology of preparation microbial inoculum: unfavorable needle wood chip material is carried out preliminary treatment; Select collocation for reducing cost with the abundant material in locality, adjust, make this practical new technology can obtain promotion and application widely by selection proportioning and dosage to the various nutrient components that adapt with it.
The present invention can suit measures to local conditions, gather materials on the spot, because of leaking just letter, processed and applied on the spot, will further strengthen the adaptability of microbial inoculum, and can extensive use in the locality.
Embodiment
The present invention is described in further detail below in conjunction with embodiment:
Embodiment 1. adopts the microbial inoculum (be mainly used in the Da Xiaoxinanlin Mountains) of larch wood chip as the matrix preparation
Its preparation process is as follows:
(1) substrate composition: get that larch wood chip 70% steam steams in advance, atmosphere gas discharging 2 hours, make the aromatic substance volatilization in the matrix, differentiate it with the aromatic odor weight, after get wheat bran 30% and mix by weight with the larch wood chip;
(2) nutrient solution is mixed matrix: interpolation diammonium hydrogen phosphate 0.2g, glucose 5g, calcium chloride 0.05g, sodium chloride 0.05g, potassium dihydrogen phosphate 3g, peptone 1.0g, magnesium sulfate 0.05g, ferric trichloride 0.004g, zinc sulphate concentration are that 10ppm, sodium molybdate concentration are 10ppm and Cobastab in 1 liter water
10.05g, make water content of substrate reach 65%;
(3) sterilization: the matrix that will mix, the solid-liquid proportioning is suitable, bottling is sealed, and carries out water vapour at 110 ℃ under a little more than an atmospheric pressure and sterilizes 2 hours, cools off standby;
(4) thalline is made: (slant culture) bacterial classification and medium were inoculated with 2: 100 ratios after one-level was cultivated, with biological fermentation process production (shaking table shaken cultivation), specifically get the sliding rust of burr umbrella (Hebeloma mesophaeum), ash ring Suillus bovines (Suillus aeruginascens) and reddish brown cortina bacterium (Cortinarius russus), in 1 liter water, add: potato 100g, glucose 10g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.5g, the plain B of little life
10.02g, make medium, be that 100ml is sub-packed in the triangular flask of 500ml with the volume, with shaking table vibration constant temperature culture 3 days;
(5) microbial inoculum: the thalline produced is inoculated in the above-mentioned solid matrix 28 ℃ of constant temperature culture 15 days with 7% amount.
The microbial inoculum (being mainly used in cool temperate zone and north temperate zone) that embodiment 2. adopts perlite, the peat composed of rotten mosses to prepare as matrix
(1) substrate composition: birch bits 25%, perlite 40%, the peat composed of rotten mosses 20%, wheat bran 15% are mixed by weight;
(2) nutrient solution is mixed matrix: add diammonium hydrogen phosphate 0.3g, glucose 10.0g, peptone 3.0g, calcium chloride 0.08g, sodium chloride 0.2g, potassium dihydrogen phosphate 2.0g, magnesium sulfate 0.01g, ferric trichloride 0.005g and Cobastab with 1 liter water
10.04g, make water content of substrate reach 70%;
(3) sterilization: will mix the suitable matrix of solid-liquid proportioning and pack and seal, and carry out steam sterilizing 1.5 hours under a little more than an atmospheric pressure, and cool off standby at 115 ℃;
(4) thalline is made: one-level is cultivated the back bacterial classification mix with 5: 100 ratios with medium, with biological fermentation process production (shaking table shaken cultivation), specifically get kakgeusia breast mushroom (Lactarius insulsus), burr sliding rust umbrella (Hebeloma mesophaeum) and reddish brown cortina bacterium (Cortinarius russus); In 1 liter water, add: potato 300g, glucose 20g, potassium dihydrogen phosphate 3g, magnesium sulfate 1.5g, the plain B of little life
10.05g, be made into medium, be that 130ml is sub-packed in the 500ml triangular flask with the volume, shaking table vibration constant temperature culture 5 days;
(5) microbial inoculum: the thalline produced is inoculated in the above-mentioned solid matrix 23 ℃ of constant temperature culture 30 days with 10% amount.
Embodiment 3. adopts the microbial inoculum (be mainly used in Guangxi Yulin Prefecture) of masson pine wood chip as the matrix preparation
(1) masson pine wood chip preliminary treatment; Get that 120 degrees centigrade of 60% masson pine wood chip water vapours are pre-to steam gas discharging after 30 minutes so that the volatilization of the aromatic substance in the matrix is differentiated it with the aromatic odor weight, after get wheat bran 40% and mix by weight with the masson pine wood chip;
(2) nutrient solution is mixed matrix: in 1 liter water, add diammonium hydrogen phosphate 0.1g, glucose 4.0g, calcium chloride 0.03g, sodium chloride 0.1g, potassium dihydrogen phosphate 3.0g, pure female cream 0.2g, magnesium sulfate 0.1g, ferric trichloride 0.001g, the plain B of little life
10.04 zinc sulphate concentration is that 20ppm, sodium molybdate concentration are 30ppm, makes water content of substrate reach 60%;
(3) sterilization: suitable matrix bottling is sealed will to mix the solid-liquid proportioning, carries out water vapour at 113 ℃ under a little more than an atmospheric pressure and sterilizes 1.8 hours, cools off standby;
(4) thalline is made: bacterial classification and medium were inoculated with 4: 100 ratios after one-level was cultivated, with biological fermentation process production (shaking table shaken cultivation), specifically get reddish brown cortina bacterium (Cortinarius russus), the sliding rust of burr umbrella (Hebeloma mesophaeum), kakgeusia breast mushroom (Lactarius insulsus) and high handle mushroom (Lepiota procera); In 1 liter water, add medium: potato 200g, glucose 15g, brewer's wort 4 Baume degrees, potassium dihydrogen phosphate 4g, magnesium sulfate 2.5g, the plain B of little life
10.04g, make medium, be that 120ml is sub-packed in the 500ml triangular flask with the volume, shaking table vibration constant temperature culture 7 days;
(5) microbial inoculum: the thalline produced is inoculated in the above-mentioned solid matrix with 5% amount, and constant temperature was cultivated 20 days for 27 ℃.
Embodiment 4. adopts the microbial inoculum (be mainly used in non-forest zone) of agricultural byproducts offcuts as the matrix preparation
(1) substrate composition: get maple wood chip 30%, rice husk 45%, straw 15%, wheat bran 10% by weight mixing;
(2) nutrient solution is mixed matrix: to add diammonium hydrogen phosphate 0.4g, glucose 8g, calcium chloride 0.01g, sodium chloride 0.3g, peptone 2.0g, yeast extract 0.3g, potassium dihydrogen phosphate 1g, dipotassium hydrogen phosphate 2g, magnesium sulfate 0.2g in the 1 liter water, ferrous sulfate concentration is 50ppm, ferric trichloride 0.002g and Cobastab
10.02g, make water content of substrate reach 50%;
(3) sterilization: suitable matrix pack or bottle sealed will to mix the solid-liquid proportioning, carries out water vapour sterilization 1.0 hours under 121 ℃ of 1.5 atmospheric pressure, cools off standby;
(4) thalline is made: bacterial classification and medium after one-level is cultivated are inoculated with 3: 100 ratios, with biological fermentation process production (shaking table shaken cultivation).Specifically get the sliding rust of burr umbrella (Hebelomamesophaeum), kakgeusia breast mushroom (Lactarius insulsus) and pinecone beef liver (Boletusfloccopus); In 1 liter water, add: potato 100g, glucose 18g, potassium dihydrogen phosphate 5g, magnesium sulfate 2g, the plain B of little life
10.03g, make medium, be that 140ml is sub-packed in the 500ml triangular flask with the volume, shaking table shaken cultivation 5 days;
(5) microbial inoculum: the thalline produced is inoculated in the above-mentioned solid matrix with 4% amount, cultivated 25 days for 25 ℃.
Utilize microbial inoculum of the present invention, in being applied to Daxing'an Mountainrange burned area forest resources recovery and upgrading, the Mycorrhizal nursery stock of cultivating is afforested on burned area, and survival rate reaches more than 95% then; High then growth, the Dahurian larch comparison is shone and is improved 128% according to raising 80%, pinus sylvestris var. mongolica comparison.
Utilize microbial inoculum of the present invention, the masson pine in state-run 60,000 forest farms, Yulin Prefecture, Guangxi is grown seedlings change once stubble for 2-3, be the forest farm 400-500 unit/mu of cutting down expenses by original 1 year one change of crops court of a feudal ruler length.
Claims (7)
1. the preparation method of the area forest mycorrhizal fungi microbial inoculum different with raw material sources is characterized in that can preparing according to the following procedure by weight each component of mixed preparing:
1) the matrix collocation mixes: matrix comprises that 3-4 part wood chip and 1-2 part wheat bran mix;
2) nutrient solution preparation: by adding diammonium hydrogen phosphate 0.1-0.5g, glucose 2-10g, calcium chloride 0.01-0.08g, magnesium sulfate 0.01-0.2g, ferric trichloride 0.001-0.005g, sodium chloride 0.05-0.3g, Cobastab in the 1 liter water
10.01-0.05g be made into nutrient solution with potassium dihydrogen phosphate 0.5-5g and/or dipotassium hydrogen phosphate 0.32-3.2g;
3) sterilization: above-mentioned nutrient solution and matrix are puddled, and making the water content of substrate after puddling is 50-70%, and packing is sealed, and adds to depress at 105-125 ℃ and carries out steam sterilizing 1-2 hour, cools off standby;
4) thalline preparation: bacterial classification and medium after the one-level of learning from else's experience is cultivated, be 2-5 with bacterial classification with the mixed proportion of medium: 100 mix, and produce thalline with the two stage biological fermentation method;
5) fungicide preparation: the matrix after will sterilizing was inoculated with the biomass of its 2-10%, at 23-30 ℃ of following constant temperature culture 10-30 days.
2. according to the preparation method of the described forest mycorrhizal fungi of claim 1 microbial inoculum, it is characterized in that: also can add the adding of 10-70% that accounts for the matrix gross weight in the described matrix and fill agent, add fill agent be in the peat composed of rotten mosses, wheat tangerine, straw, rice husk, the corn straw one or more.
3. according to the preparation method of the described forest mycorrhizal fungi of claim 1 microbial inoculum, it is characterized in that: also can add in the described matrix and account for matrix gross weight 10-50% perlite as carrier
4. according to the preparation method of the described forest mycorrhizal fungi of claim 1 microbial inoculum, it is characterized in that: also can select to add one or more following materials in the described nutrient solution preparation: yeast extract 0.05-0.3g, peptone 1.0-3.0g, ferrous sulfate concentration is 10-50ppm, zinc sulphate concentration is 10-50ppm, and sodium molybdate concentration is 10-50ppm.
5. according to the preparation method of the described forest mycorrhizal fungi of claim 1 microbial inoculum, it is characterized in that: described wood chip is one or more mixing in birch bits, larch wood chip, maple bits, the masson pine wood chip.
6. according to the preparation method of claim 1 or 5 described forest mycorrhizal fungi microbial inoculums, it is characterized in that: need before mixed substrate, to steam more than 30 minutes in advance when described wood chip is the needle wood chip or normal pressure steams more than 2 hours in advance, carry out gas simultaneously and discharge with 120 degrees centigrade of steam.
7. according to the preparation method of the described forest mycorrhizal fungi of claim 1 microbial inoculum, it is characterized in that: described bacterial classification is the forest mycorrhizal fungi: one or more mixing in kakgeusia breast mushroom, the sliding rust of burr umbrella, ash ring Suillus bovines, pinecone beef liver, high handle mushroom, the reddish brown cortina bacterium.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103814756A (en) * | 2014-03-07 | 2014-05-28 | 深圳市万信达生态环境股份有限公司 | Method suitable for quick rooting and surviving of southern landscape tree after transplantation |
| CN104892095A (en) * | 2015-05-08 | 2015-09-09 | 云南省热带作物科学研究所 | Mould-inhibition and yield-increasing panacea dedicated to soil covering process for Phlebopus portentosus |
| CN106455581A (en) * | 2014-06-06 | 2017-02-22 | 拉斯夫雷公司 | Method and composition for improving the productivity of non-leguminous plants |
| CN107162753A (en) * | 2017-06-14 | 2017-09-15 | 广东东阳光药业有限公司 | The culture medium for cultivating and method of Phlebopus portentosus |
| CN108004157A (en) * | 2017-11-15 | 2018-05-08 | 潍坊友容实业有限公司 | The preparation method and application method of salt-soda soil crop-planting compound nutritional microbial inoculum, the microbial inoculum |
| CN108624512A (en) * | 2018-05-30 | 2018-10-09 | 内蒙古和盛生态科技研究院有限公司 | Solid fermentation matrix, preparation method and the method for cultivating mycorhiza biological agent |
| CN114535286A (en) * | 2022-02-24 | 2022-05-27 | 东北林业大学 | Method for repairing heavy metal contaminated soil by using ectomycorrhizal fungi and dahurian larch |
| CN117256406A (en) * | 2023-10-23 | 2023-12-22 | 贵州省生物研究所 | Convenient artificial preparation method of fresh Lactarius deliciosus mycorrhizal seedlings |
-
2001
- 2001-09-17 CN CN 01128174 patent/CN1236049C/en not_active Expired - Fee Related
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103814756A (en) * | 2014-03-07 | 2014-05-28 | 深圳市万信达生态环境股份有限公司 | Method suitable for quick rooting and surviving of southern landscape tree after transplantation |
| CN106455581A (en) * | 2014-06-06 | 2017-02-22 | 拉斯夫雷公司 | Method and composition for improving the productivity of non-leguminous plants |
| CN104892095A (en) * | 2015-05-08 | 2015-09-09 | 云南省热带作物科学研究所 | Mould-inhibition and yield-increasing panacea dedicated to soil covering process for Phlebopus portentosus |
| CN107162753A (en) * | 2017-06-14 | 2017-09-15 | 广东东阳光药业有限公司 | The culture medium for cultivating and method of Phlebopus portentosus |
| CN107162753B (en) * | 2017-06-14 | 2020-08-11 | 东莞东阳光保健品研发有限公司 | Culture medium and method for phlebopus portentosus |
| CN108004157A (en) * | 2017-11-15 | 2018-05-08 | 潍坊友容实业有限公司 | The preparation method and application method of salt-soda soil crop-planting compound nutritional microbial inoculum, the microbial inoculum |
| CN108624512A (en) * | 2018-05-30 | 2018-10-09 | 内蒙古和盛生态科技研究院有限公司 | Solid fermentation matrix, preparation method and the method for cultivating mycorhiza biological agent |
| CN114535286A (en) * | 2022-02-24 | 2022-05-27 | 东北林业大学 | Method for repairing heavy metal contaminated soil by using ectomycorrhizal fungi and dahurian larch |
| CN117256406A (en) * | 2023-10-23 | 2023-12-22 | 贵州省生物研究所 | Convenient artificial preparation method of fresh Lactarius deliciosus mycorrhizal seedlings |
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