CN1236049C - 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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- CN1236049C CN1236049C CN 01128174 CN01128174A CN1236049C CN 1236049 C CN1236049 C CN 1236049C CN 01128174 CN01128174 CN 01128174 CN 01128174 A CN01128174 A CN 01128174A CN 1236049 C CN1236049 C CN 1236049C
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Abstract
The present invention relates to a preparation method of microbial inocula of tree ectomycorrhiza fungi from different areas and feed sources. Substrate wood dust, wheat bran, pearl stone, grass carbon, etc. are selectively matched and mixed according to a proportion by weight, inorganic salts and partial nutritive elements are added, nutrient solution whose water content of substrates is from 50 to 70% is prepared and used for mixing materials, material liquid is packaged and sealed after being evenly stirred, and then the material liquid is sterilized by pressurized steam; 2 to 10 wt% of thalli which are produced in advance by secondary biofermentation are inoculated into the substrates after being cooled, and the thalli are cultured for 10 to 30 days at a constant temperature. The present invention is free from the limitation of feed sources and areas, the cost is low, and the present invention is suitable for generalization and application.
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 nature fungi and plant symbiosis.Mycorhiza can promote the absorption of plant to moisture and nutritive substance, produces biologically active substance, strengthens the resistance of plant, and under certain plants or particular environment condition, the existence of mycorhiza can become the determinative 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 inoculation, 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 environment protection does a lot of work, and obtained unusual effect.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 tankage 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) nutritive medium preparation: by adding Secondary ammonium phosphate 0.1-0.5g, glucose 2-10g, calcium chloride 0.01-0.08g, sal epsom 0.01-0.2g, iron trichloride 0.001-0.005g, sodium-chlor 0.05-0.3g, vitamins B in the 1 liter water
10.01-0.05g be made into nutritive medium with potassium primary phosphate 0.5-5g and/or dipotassium hydrogen phosphate 0.32-3.2g;
3) sterilization: above-mentioned nutritive medium 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 substratum after the one-level of learning from else's experience is cultivated, be 2-5 with bacterial classification with the blending ratio of substratum: 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 stalk 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 nutritive medium preparation: yeast extract paste 0.05-0.3g, peptone 1.0-3.0g, ferrous sulfate concentration is 10-50ppm, and zinc sulfate concentration is 10-50ppm, and Sodium orthomolybdate concentration is 10-50ppm;
Described wood chip is one or more mixing in birch bits, tamarack wood chip, maple bits, the Pinus massoniana Lamb 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 gaseous emission simultaneously 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 working process---wood chip, tankage of farm crop (as wheat tangerine, straw, rice husk, corn stalk) and material of construction 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 tankage 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 geographic restriction.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, geographic difference, 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 pre-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 nutritive ingredients 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 widespread 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 tamarack wood chip as the matrix preparation
Its preparation process is as follows:
(1) substrate composition: get that tamarack wood chip 70% steam steams in advance, atmosphere gas discharging 2 hours, make the aromatic essence volatilization in the matrix, differentiate it with the aromatic odour weight, after get wheat bran 30% and mix by weight with the tamarack wood chip;
(2) nutritive medium is mixed matrix: interpolation Secondary ammonium phosphate 0.2g, glucose 5g, calcium chloride 0.05g, sodium-chlor 0.05g, potassium primary phosphate 3g, peptone 1.0g, sal epsom 0.05g, iron trichloride 0.004g, zinc sulfate concentration are that 10ppm, Sodium orthomolybdate concentration are 10ppm and vitamins B 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 vapor at 110 ℃ under a little more than a normal atmosphere and sterilizes 2 hours, cools off standby;
(4) thalline is made: (slant culture) bacterial classification and substratum were inoculated with 2: 100 ratios after one-level was cultivated, with biological fermentation process production (shaking table shaking culture), 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 primary phosphate 1g, sal epsom 0.5g, the plain B of little life
10.02g, make substratum, 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 substrate 28 ℃ of constant temperature culture 15 days with 7% amount.
The microbial inoculum (being mainly used in cool temperature 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) nutritive medium is mixed matrix: add Secondary ammonium phosphate 0.3g, glucose 10.0g, peptone 3.0g, calcium chloride 0.08g, sodium-chlor 0.2g, potassium primary phosphate 2.0g, sal epsom 0.01g, iron trichloride 0.005g and vitamins B 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 a normal atmosphere, and cool off standby at 115 ℃;
(4) thalline is made: bacterial classification and substratum were with 5: 100 mixed after one-level was cultivated, with biological fermentation process production (shaking table shaking culture), 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 primary phosphate 3g, sal epsom 1.5g, the plain B of little life
10.05g, be made into substratum, 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 substrate 23 ℃ of constant temperature culture 30 days with 10% amount.
Embodiment 3. adopts the microbial inoculum (be mainly used in Guangxi Yulin Prefecture) of Pinus massoniana Lamb wood chip as the matrix preparation
(1) Pinus massoniana Lamb wood chip pre-treatment; Get that 120 degrees centigrade of 60% Pinus massoniana Lamb wood chip water vapors are pre-to steam gaseous emission after 30 minutes so that the volatilization of the aromatic essence in the matrix is differentiated it with the aromatic odour weight, after get wheat bran 40% and mix by weight with the Pinus massoniana Lamb wood chip;
(2) nutritive medium is mixed matrix: in 1 liter water, add Secondary ammonium phosphate 0.1g, glucose 4.0g, calcium chloride 0.03g, sodium-chlor 0.1g, potassium primary phosphate 3.0g, yeast extract paste 0.2g, sal epsom 0.1g, iron trichloride 0.001g, the plain B of little life
10.04 zinc sulfate concentration is that 20ppm, Sodium orthomolybdate 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 vapor at 113 ℃ under a little more than a normal atmosphere and sterilizes 1.8 hours, cools off standby;
(4) thalline is made: bacterial classification and substratum were inoculated with 4: 100 ratios after one-level was cultivated, with biological fermentation process production (shaking table shaking culture), 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 substratum: potato 200g, glucose 15g, wort 4 degree Beaume, potassium primary phosphate 4g, sal epsom 2.5g, the plain B of little life
10.04g, make substratum, 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 substrate 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 tankage as the matrix preparation
(1) substrate composition: get maple wood chip 30%, rice husk 45%, straw 15%, wheat bran 10% by weight mixing;
(2) nutritive medium is mixed matrix: to add Secondary ammonium phosphate 0.4g, glucose 8g, calcium chloride 0.01g, sodium-chlor 0.3g, peptone 2.0g, yeast extract paste 0.3g, potassium primary phosphate 1g, dipotassium hydrogen phosphate 2g, sal epsom 0.2g in the 1 liter water, ferrous sulfate concentration is 50ppm, iron trichloride 0.002g and vitamins B
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 vapor sterilization 1.0 hours under 121 ℃ of 1.5 normal atmosphere, cools off standby;
(4) thalline is made: bacterial classification and substratum after one-level is cultivated are inoculated with 3: 100 ratios, with biological fermentation process production (shaking table shaking culture).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 primary phosphate 5g, sal epsom 2g, the plain B of little life
10.03g, make substratum, be that 140ml is sub-packed in the 500ml triangular flask with the volume, shaking table shaking culture 5 days;
(5) microbial inoculum: the thalline produced is inoculated in the above-mentioned solid substrate with 4% amount, cultivated 25 days for 25 ℃.
Utilize microbial inoculum of the present invention, in being applied to Daxing'an Mountainrange brulee forest reserves recovery and upgrading, the Mycorrhizal nursery stock of cultivating is afforested on brulee, and surviving rate reaches more than 95% then; High then growth, the larch in Xinanlin area comparison is shone and is improved 128% according to raising 80%, pinus sylvestris var. mongolica comparison.Utilize microbial inoculum of the present invention, the Pinus massoniana Lamb 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 by weight each component of mixed preparing, according to the following procedure preparation:
1) the matrix collocation mixes: matrix comprises that 3-4 part wood chip and 1-2 part wheat bran mix;
2) nutritive medium preparation: by adding Secondary ammonium phosphate 0.1-0.5g, glucose 2-10g, calcium chloride 0.01-0.08g, sal epsom 0.01-0.2g, iron trichloride 0.001-0.005g, sodium-chlor 0.05-0.3g, vitamins B in the 1 liter water
10.01-0.05g be made into nutritive medium with potassium primary phosphate 0.5-5g and/or dipotassium hydrogen phosphate 0.32-3.2g;
3) sterilization: above-mentioned nutritive medium and matrix are puddled, and making the water content of substrate after puddling is 50-70%, and packing is sealed, and carries out under 105-125 ℃ steam sterilizing 1-2 hour, cools off standby;
4) thalline preparation: bacterial classification and substratum after the one-level of learning from else's experience is cultivated, be 2-5 with bacterial classification with the blending ratio of substratum: 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 stalk 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 nutritive medium preparation: yeast extract paste 0.05-0.3g, peptone 1.0-3.0g, ferrous sulfate concentration is 10-50ppm, zinc sulfate concentration is 10-50ppm, and Sodium orthomolybdate 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, tamarack wood chip, maple bits, the Pinus massoniana Lamb 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 when described wood chip is the needle wood chip before mixed substrate, to steam 2 hours in advance, carry out gaseous emission simultaneously with 120 degrees centigrade of pre-steamings of steam 30 minutes or normal pressure.
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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| CN 01128174 CN1236049C (en) | 2001-09-17 | 2001-09-17 | Method for preparing forest mycorrhizal fungi preparation from different area and raw material |
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| CN 01128174 CN1236049C (en) | 2001-09-17 | 2001-09-17 | Method for preparing forest mycorrhizal fungi preparation from different area and raw material |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| 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 |
| FR3021843B1 (en) * | 2014-06-06 | 2016-07-08 | Lesaffre & Cie | 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 |
| 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 |
| CN108624512B (en) * | 2018-05-30 | 2021-03-30 | 内蒙古和盛生态科技研究院有限公司 | Solid fermentation substrate, preparation method and method for culturing mycorrhiza biological preparation |
| 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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