CN103070014A - Method for breeding summer truffle root seedling through inoculation of suspension liquid - Google Patents
Method for breeding summer truffle root seedling through inoculation of suspension liquid Download PDFInfo
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Abstract
The invention discloses a method for breeding summer truffle root seedling through inoculation of suspension liquid, which is characterized in that the difficulty in artificial inoculation of summer truffle can be solved through steps of breeding of aseptic seedling, preparation of microbial inoculum, inoculation of suspension liquid, seedling exercising, breeding of fungus seedlings and test and confirmation. Compared with the existing breeding method, the method is characterized in that pinus armandi, castanea mollissima and pinus yunnanensis are adopted as hosts; under a relatively aseptic condition, i.e. the condition for preventing the competition of fungi and infection of most mycetes, the summer truffle spores sufficiently contact with roots of the host trees; the single infection predominance of the summer truffle is maintained within a given period of time; and a great number of firm summer truffle mycorrhizas can be formed.
Description
Technical field
The present invention relates to a kind of Va Mycorrhiza Seedling artificial infection breeding method, relate in particular to the method that the summer truffle offspring is cultivated in a kind of suspension inoculation.
Background technology
Ferfas usually and fine-root have symbiotic relation, be important wild edible fungus, its ascocarp contains higher nutrient component, commercial value is attracted attention by common people.French, gondola black truffle (Tuber melanosporum) and white ferfas (Tuber magnatum) be price such as platinum in the international market, and become the important component part of local culture.(the Latin formal name used at school: tube aestivum vitt.), ascocarp is irregular sphere to summer truffle, and base portion often has depression, diameter 3-6cm, it is protruding that the surface has the dihedral wart on 5-6 limit, and the diameter of wart can reach 0.5cm, wants surface characteristics obvious than black truffle and India truffle ascus.Inner (spore body) pale asphyxia, after be cream-colored or isabelline, in the train of thought of mutual commissure and branch is arranged.
Domestic more than the 20 kind ferfas kinds such as India truffle, summer truffle, white ferfas that had been found that at present, mainly be distributed in the Southwestern China each province, produce and be annual in February, the 2nd in October 1 season, white is to canescence during the children, maturation is black or brown, has clearly milky vein, and that eats is delicious, contain rich in protein and multivitamin, especially vitamin A, vitamin B2 content are high.Because the growth of ferfas must be sought commensalism with suitable host plant root system, otherwise can't form ascocarp, this just means that the artificial cultivation ferfas be unable to do without the live body host plant, thereby the ferfas artificial cultivation is relatively more difficult, with high content of technology, realize that the artificial cultivation of ferfas must at first break through the cultivation technology in mycorhiza south.Although domestic Tuber Sinense and India truffle to China is more in the research aspect the physiological ecological, the first synthetic cultivation technology of its truffle mycorrhiza fungi also there are Primary Study and report.But technology is too much in laboratory operation, and production operation is not strong, and summer truffle mycorrhiza fungi seeding cultivating technology rarely has and appears in the newspapers.
The patented invention " a kind of ferfas and mycorhiza synthetic method " that approaches, number of patent application 200810058447.9
Existing problems one: " a kind of ferfas and mycorhiza synthetic method " is single for the examination seeds, and the ferfas bacterial classification is single.Only relate to coniferous species in the method, do not related to deciduous species; Only related to Tuber Sinense in the method, namely India truffle does not relate to the higher summer truffle of internationally recognized value, white ferfas.
The problem two that exists: cultivating aseptic seedling.Aseptic seedling matrix adopts sterilization 1 hour perlite and vermiculite (1/1 mixes) in " a kind of ferfas and mycorhiza synthetic method ", can guarantee the water penetration in the cultivating seedlings, but can not solve the nutrient nutritional need problem of seedling growth, seedling growth is slow, and the matrix sterilization time is inadequate, occurs easily polluting.And cultivating aseptic seedling should cultivate in the greenhouse of overhead relative closure, could ensure not to be subjected to especially infecting of fungi of other miscellaneous bacteria in the cultivating aseptic seedling process.
The problem three that exists: inoculate matrix formulations in " a kind of ferfas and mycorhiza synthetic method " and be humus, water, vermiculite 1/1/1 and mix, pH value adjusts to 7.0.In fact the substrate composition otherness is larger between different tree species, and with regard to the Huashan pine Va Mycorrhiza Seedling of India truffle, sieve laterite and vermiculite are much better by 1/1 mixed effect, and the adjustment of P in soil H value should be in the interval range of 6.5-7.0.
The problem four that exists: the preparation of " a kind of ferfas and mycorhiza synthetic method " microbial inoculum and inoculation technique nursery operability are not strong, easily cause 2 pollutions.The one, after the ferfas ascocarp took out, not to its surface sterilizing, other fungi and earth that its ascocarp surface may be contained can not be washed off by sterile water, may cause the direct pollution of Va Mycorrhiza Seedling.The 2nd, what vaccination ways was taked is to dip in root, and the concentration of inoculation liquid is also less than normal, dips in the process because root system development etc. are inconsistent, will cause between each seedling of inoculation the spore amount asymmetric.
The problem five that exists: " a kind of ferfas and mycorhiza synthetic method " lacks the hardening process.Mycorrhiza fungi seeding cultivating is directly to be placed in the booth of gravity-flow ventilation to carry out in " a kind of ferfas and mycorhiza synthetic method ", initial 15-30 days of cultivating seedlings, need the aseptic space of space to carry out hardening, truffle mycorrhiza fungi forms effect can be more stable, and the initial stage of stopping is subjected to the competitive influence of extraneous miscellaneous bacteria.
Summary of the invention
Purpose of the present invention just is that providing a kind of addresses the above problem, for the artificial infection breeding method of summer truffle Va Mycorrhiza Seedling.
To achieve these goals, the technical solution used in the present invention is: the method that the summer truffle offspring is cultivated in a kind of suspension inoculation, comprise the steps,
(1) cultivate the host plant seedling, specific as follows,
A, gather, select the host plant seed;
Described host plant is selected a kind of of pinus yunnanensis, Huashan pine, Chinese chestnut;
B, processing host plant seed;
The seed wet concentration that steps A is collected goes out ghost, the H with 30%
2O
2After the immersion treatment 40 minutes, use distilled water flushing, until without H
2O
2Till the smell;
C, preparation medium;
With vermiculite and peat soil by volume 1:1 mix to pack into and grow seedlings in the frame, through high temperature 121-126 ℃ of high pressure 1.5kg/cm
2Sterilization is more than 2 hours, forms the aseptic substrate frame of growing seedlings;
D, cultivation host plant;
Plant seed after step B processes in time is sown into aseptic substrate grows seedlings in the frame, then be positioned over vernalization in the greenhouse, the rear immigration booth that sprouts is educated strain, until grow offspring;
(2) the preparation microbial inoculum is specific as follows,
A, Collection and preservation bacterial classification;
Gather the summer truffle fruit body, clean gently surperficial earth with brush body first, in 75% alcohol, soak again 3min to its surface sterilizing, simultaneously more fast calcination on alcolhol burner flame, continue sterilization, then place the river sand behind autoclave sterilization, stored refrigerated is for subsequent use under 3-5 ℃ temperature, and described river sand adopts 121-126 ℃ of high pressure 1.5kg/cm of high temperature
2Sterilization is more than 2 hours;
B, preparation microbial inoculum; Its compound method is as follows,
The first step, take by weighing first a certain amount of ferfas, with soaking 1-3 minute in 75% the alcohol to its surface sterilizing, and then fast calcination on alcolhol burner flame, continue sterilization, place on the operating desk after the sterilization, with the beater after the alcohol sterilization it is ground into particle diameter less than lmm suspension, make the spore of bacterial classification go out shell, ferfas and distilled water volume ratio are 1:5 in the suspension;
Second step is with high-hydroscopicity acrylic resin and the by volume 1:400 preparation of ferfas suspension;
The 3rd step is with the distilled water solution of the instant rear preparation 60PPM concentration of heteroauxin;
In the 4th step, the suspension that second step is made and the distilled water solution of the 3rd step making by volume 1:1 are mixed with the ferfas Inoculant;
(3) the suspension inoculation method is adopted in inoculation, and is specific as follows,
The host plant seedling that A, arrangement cultivate deducts unnecessary root, leaf;
B, insert the sterile soil through disinfection in inoculation cup, the sterile soil amount of fill accounts for 1/3 of inoculation cup volume, the host plant seedling is put into cup central authorities again;
C, sprinkle ferfas suspension microbial inoculum to host plant shoot root section, make it to be attached on the root system;
D, filling inoculation culture medium cover root system;
(4) hardening;
The plant that rigidly connects kind of ferfas is positioned over first temperature between 15-25 ℃, and the humidity of matrix remains in the seeding room of 50-80% and cultivated 15-30 days on the seedling culture frame, makes bacterial classification parasitic success on the host;
(5) cultivate Va Mycorrhiza Seedling;
Parasitic successful plant is moved out on the seedling culture frame of glass booth and cultivated 6-24 month, become the commercial seedling that can supply field production.
As preferably, among the step C of described cultivating aseptic seedling, described pinus yunnanensis, the uniform broadcasting of Huashan pine seed are in the frame of growing seedlings, on cover the 1cm sterilization matrix, and Castanea Mollissima Seeds adopts program request in the frame of growing seedlings, on cover the 2-3cm sterilization matrix, be positioned over after with clear water matrix being irrigated in the greenhouse of 25 ℃ of temperature and carry out vernalization, move into temperature again be behind the seed sprouting more than 80% in 10-35 ℃ the booth and carry out plant cultivating, the humidity with matrix remains on 50-80% simultaneously.
As preferably, in the described microbial inoculum preparation process, the general storage life of bacterial classification is 1-3 month; If long preservation will place-18 ℃ the interior vacuum of refrigerating chamber to preserve.
As preferably, among the described inoculation step D, described inoculation culture medium uses 121-126 ℃ of high pressure 1.5kg/cm of high temperature
2The aseptic substrate of sterilization more than 2 hours, according to the difference of seeds, the peat soil that pinus yunnanensis should be adopted and vermiculite ratio by volume 1:1 are mixed; The naturally native and vermiculite ratio that Huashan pine and Chinese chestnut should be adopted by volume 1:1 is mixed; The pH value of matrix is regulated with quicklime, maintains 7.5.
As preferably, described method of selecting the host plant seed is, pinus yunnanensis and Huashan pine are removed with wet concentration and dry rear normal temperature after the ghost and be in store for; The Castanea Mollissima Seeds wet concentration remove to go rotten and not full seed, dries surface moisture, again with humidity be 25-35% clean river sand by volume the ratio of 1:2 mix, and place 5 ℃ refrigerator to save backup.
As preferably, among the step C of described artificial infection, described inoculum concentration 2-5g/ strain.
Compared with prior art, the invention has the advantages that: a difficult problem that the invention solves the summer truffle artificial infection, take Huashan pine, Chinese chestnut, pinus yunnanensis as the host, with the summer truffle spore under relatively aseptic condition, namely stop under the competition of fungi and the condition that most mould is infected, capacity contacts with summer truffle host seeds root, keeps within a certain period of time the single infection advantage of summer truffle spore, forms firm and a large amount of summer truffle mycorhiza.The inoculation of employing suspension, the suspension that has added super absorbent resin can be adsorbed on the spore that suspends in the liquid fully, guarantee the uniformity that spore suspension distributes, and can more effectively be adsorbed on the root of plant, and can both play water retention at cultivating seedlings with when going out to plant.And increase low concentration auxin heteroauxin, can stimulate on the quality and quantity of plant root growth and fibrous root and increase, improve the quantity of inoculation efficiency and the formation mycorhiza of ferfas spore, the suspension inoculation method is few to the fibrous root growth, and the conifer plant is remarkable especially slowly.
Description of drawings
Fig. 1 is schematic flow sheet of the present invention.
Embodiment
The invention will be further described below in conjunction with accompanying drawing.
Embodiment: referring to Fig. 1, the method that the summer truffle offspring is cultivated in a kind of suspension inoculation comprises following processing step,
(1) cultivate the host plant seedling, specific as follows,
A, gather, select the host plant seed;
Described host plant is selected a kind of of pinus yunnanensis, Huashan pine, Chinese chestnut; Described method of selecting the host plant seed is, pinus yunnanensis and Huashan pine are removed with wet concentration and dry rear normal temperature after the ghost and be in store for; The Castanea Mollissima Seeds wet concentration remove to go rotten and not full seed, dries surface moisture, again with humidity be 25-35% clean river sand by volume the ratio of 1:2 mix, and place 5 ℃ refrigerator to save backup;
B, processing host plant seed;
The seed wet concentration that steps A is collected goes out ghost, the H with 30%
2O
2After the immersion treatment 40 minutes, use distilled water flushing, until without H
2O
2Till the smell;
C, preparation medium;
With vermiculite and peat soil by volume 1:1 mix to pack into and grow seedlings in the frame, through high temperature 121-126 ℃ of high pressure 1.5kg/cm
2Sterilization is more than 2 hours, forms the aseptic substrate frame of growing seedlings;
D, cultivation host plant;
Plant seed after step B processes in time is sown into aseptic substrate grows seedlings in the frame, then be positioned over vernalization in the greenhouse, the rear immigration booth that sprouts is educated strain, until grow offspring;
Described pinus yunnanensis, the uniform broadcasting of Huashan pine seed, on cover the 1cm sterilization matrix, the Castanea Mollissima Seeds program request is in the frame of growing seedlings, on cover the 2-3cm sterilization matrix, be positioned over after with clear water matrix being irrigated in the greenhouse of 25 ℃ of temperature and carry out vernalization, move into temperature again be behind the seed sprouting more than 80% in 10-35 ℃ the booth and carry out plant cultivating, the humidity with matrix remains on 50-80% simultaneously.
(2) the preparation microbial inoculum is specific as follows,
A, Collection and preservation bacterial classification;
Collect the 9-12 month in the season of summer truffle maturation ripe, stature is large, external form good, not damaged, summer truffle fruit body that fragrance is relatively dense, clean gently surperficial earth with toothbrush first, in 75% alcohol, soak again 3min to its surface sterilizing, simultaneously again on alcolhol burner flame fast calcination be placed on high temperature 121-126 ℃ of high pressure 1.5kg/cm
2In the river sand of sterilization more than 2 hours, stored refrigerated is for subsequent use under 3-5 ℃ temperature; General storage life is 1-3 month; If long preservation will place-18 ℃ the interior vacuum of refrigerating chamber to preserve;
B, preparation microbial inoculum; Its compound method is as follows,
The first step, take by weighing first a certain amount of ferfas, with soaking 1-3 minute in 75% the alcohol to its surface sterilizing, and then fast calcination on alcolhol burner flame, continue sterilization, place on the operating desk after the sterilization, with the beater after the alcohol sterilization it is ground into particle diameter less than lmm suspension, make the spore of bacterial classification go out shell, ferfas and distilled water volume ratio are 1:5 in the suspension;
Second step is with high-hydroscopicity acrylic resin and the by volume 1:400 preparation of ferfas suspension;
The 3rd step is with the distilled water solution of the instant rear preparation 60PPM concentration of heteroauxin;
In the 4th step, the suspension that second step is made and the distilled water solution of the 3rd step making by volume 1:1 are mixed with the ferfas Inoculant;
The microbial inoculum of described ferfas is generally answered matching while using, if the ferfas microbial inoculum for preparing is not finished, available sealing sterilization container is temporarily deposited in 3-5 ℃ the refrigerator for subsequent use.
(3) the suspension inoculation method is adopted in inoculation, and is specific as follows,
In transfer room, lay clean stainless steel workbench, use first 75% alcohol that table-board wipe is clean after, select growing way good, the aseptic seedling of well developed root system cuts off part root system and unnecessary growth leaf with scissors, is placed on the containers for future use that sterilization treatment is crossed; Select seedling-raising cup Φ a 10 * 15cm who has used the sterilization of 0.5% liquor potassic permanganate, the sterilization soil of in cup, packing into first and accounting for cup volumes 1/3, again plant is put into cup central authorities, quantitative little spoon with 10ml is sprinkling upon the ferfas suspension for preparing on the root, add at last disinfection inoculation matrix extremely apart from 0.5cm place, seedling-raising cup edge, the inoculum concentration of ferfas microbial inoculum is the 2-5g/ strain.
The inoculation culture medium uses 121-126 ℃ of high pressure 1.5kg/cm of high temperature
2The aseptic substrate of sterilization more than 2 hours, according to the difference of seeds, the peat soil that pinus yunnanensis should be adopted and vermiculite ratio by volume 1:1 are mixed; The naturally native and vermiculite ratio that Huashan pine and Chinese chestnut should be adopted by volume 1:1 is mixed; The pH value of matrix is regulated with quicklime, and it is 7.5 more suitable generally to maintain.
(4) hardening;
The plant that rigidly connects kind of ferfas is positioned over first temperature between 15-25 ℃, and the humidity of matrix remains in the seeding room of 50-80% and cultivated 15-30 days on the seedling culture frame, makes bacterial classification parasitic success on the host;
(5) cultivate Va Mycorrhiza Seedling;
Parasitic successful plant is moved out on the seedling culture frame of glass booth and cultivated 6-24 individual month, becoming can be for the commercial seedling of field production, during mycorrhiza fungi seeding cultivating, note the health of booth periphery, in time keep the skin wet to nursery stock according to the variation of temperature and humidity; And note observing the nutrient deletion condition of plant, suitably replenish nutrient by foliar application.
(6) detect affirmation
The summer truffle inoculation can be by the microscopic features of truffle mycorrhiza fungi seeding after 3 months, assessment infection rate and infestation index; Inoculate successful truffle mycorrhiza fungi and can observe under 40-60 stereoscope doubly, that form mostly is is bar-shaped, two forked, multi-fork shape and coralliform; At the mycorhiza initial stage, the end is white in color or light golden rod yellow, and color deepens gradually and is light brown afterwards; Can clearly see that at microscopically there is translucent golden yellow mycelia on the surface, and be grown in disorderly the mycorhiza surface; Under 600 power microscopes, can be observed the sheath cell tissue that the mycorhiza top layer forms the Hartig net layer, summer truffle be sharp polygon, be different from the irregular arc decorative pattern of India truffle and black truffle formation.
The invention solves the difficult problem of summer truffle artificial infection, take Huashan pine, Chinese chestnut, pinus yunnanensis as the host, with the summer truffle spore under relatively aseptic condition, namely stop under the competition of fungi and the condition that most mould is infected, capacity contacts with summer truffle host seeds root, keep within a certain period of time the single infection advantage of summer truffle spore, form firm and a large amount of summer truffle mycorhiza.
The inoculation of employing suspension, the suspension that has added super absorbent resin can be adsorbed on the spore that suspends in the liquid fully, guarantee the uniformity that spore suspension distributes, and can more effectively be adsorbed on the root of plant, and can both play water retention at cultivating seedlings with when going out to plant.And increase low concentration auxin heteroauxin, can stimulate on the quality and quantity of plant root growth and fibrous root and increase, improve the quantity of inoculation efficiency and the formation mycorhiza of ferfas spore, the suspension inoculation method is few to the fibrous root growth, and the conifer plant is remarkable especially slowly.
More than the summer truffle offspring is cultivated in inoculation to suspension provided by the present invention method carried out detailed introduction, used specific case herein principle of the present invention and embodiment are set forth, the explanation of above embodiment just is used for helping to understand method of the present invention and core concept thereof; Simultaneously, for one of ordinary skill in the art, according to thought of the present invention, all will change in specific embodiments and applications, to change of the present invention with to improve will be possible, and can not exceed design and the scope of accessory claim defined, in sum, this description should not be construed as limitation of the present invention.
Claims (6)
1. a suspension is inoculated the method for cultivating the summer truffle offspring, it is characterized in that: comprise the steps,
(1) cultivate the host plant seedling, specific as follows,
A, gather, select the host plant seed;
Described host plant is selected a kind of of pinus yunnanensis, Huashan pine, Chinese chestnut;
B, processing host plant seed;
The seed wet concentration that steps A is collected goes out ghost, the H with 30%
2O
2After the immersion treatment 40 minutes, use distilled water flushing, until without H
2O
2Till the smell;
C, preparation medium;
With vermiculite and peat soil by volume 1:1 mix to pack into and grow seedlings in the frame, through high temperature 121-126 ℃ of high pressure 1.5kg/cm
2Sterilization is more than 2 hours, forms the aseptic substrate frame of growing seedlings;
D, cultivation host plant;
Plant seed after step B processes in time is sown into aseptic substrate grows seedlings in the frame, then be positioned over vernalization in the greenhouse, the rear immigration booth that sprouts is educated strain, until grow offspring;
(2) the preparation microbial inoculum is specific as follows,
A, Collection and preservation bacterial classification;
Gather the summer truffle fruit body, clean gently surperficial earth with brush body first, in 75% alcohol, soak again 3min to its surface sterilizing, simultaneously more fast calcination on alcolhol burner flame, continue sterilization, then place the river sand behind autoclave sterilization, stored refrigerated is for subsequent use under 3-5 ℃ temperature, and described river sand adopts 121-126 ℃ of high pressure 1.5kg/cm of high temperature
2Sterilization is more than 2 hours;
B, preparation microbial inoculum; Its compound method is as follows,
The first step, take by weighing first a certain amount of ferfas, with soaking 1-3 minute in 75% the alcohol to its surface sterilizing, and then fast calcination on alcolhol burner flame, continue sterilization, place on the operating desk after the sterilization, with the beater after the alcohol sterilization it is ground into particle diameter less than lmm suspension, make the spore of bacterial classification go out shell, ferfas and distilled water volume ratio are 1:5 in the suspension;
Second step is with high-hydroscopicity acrylic resin and the by volume 1:400 preparation of ferfas suspension;
The 3rd step is with the distilled water solution of the instant rear preparation 60PPM concentration of heteroauxin;
In the 4th step, the suspension that second step is made and the distilled water solution of the 3rd step making by volume 1:1 are mixed with the ferfas Inoculant;
(3) the suspension inoculation method is adopted in inoculation, and is specific as follows,
The host plant seedling that A, arrangement cultivate deducts unnecessary root, leaf;
B, insert the sterile soil through disinfection in inoculation cup, the sterile soil amount of fill accounts for 1/3 of inoculation cup volume, the host plant seedling is put into cup central authorities again;
C, sprinkle ferfas suspension microbial inoculum to host plant shoot root section, make it to be attached on the root system;
D, filling inoculation culture medium cover root system;
(4) hardening;
The plant that rigidly connects kind of ferfas is positioned over first temperature between 15-25 ℃, and the humidity of matrix remains in the seeding room of 50-80% and cultivated 15-30 days on the seedling culture frame, makes bacterial classification parasitic success on the host;
(5) cultivate Va Mycorrhiza Seedling;
Parasitic successful plant is moved out on the seedling culture frame of glass booth and cultivated 6-24 month, become the commercial seedling that can supply field production.
2. described a kind of suspension is inoculated the method for cultivating the summer truffle offspring according to claim 1, it is characterized in that: among the step C of described cultivating aseptic seedling, described pinus yunnanensis, the uniform broadcasting of Huashan pine seed are in the frame of growing seedlings, on cover the 1cm sterilization matrix, and Castanea Mollissima Seeds adopts program request in the frame of growing seedlings, on cover the 2-3cm sterilization matrix, be positioned over after with clear water matrix being irrigated in the greenhouse of 25 ℃ of temperature and carry out vernalization, move into temperature again be behind the seed sprouting more than 80% in 10-35 ℃ the booth and carry out plant cultivating, the humidity with matrix remains on 50-80% simultaneously.
3. described a kind of suspension is inoculated the method for cultivating the summer truffle offspring according to claim 1, it is characterized in that: in the described microbial inoculum preparation process, the general storage life of bacterial classification is 1-3 month; If long preservation will place-18 ℃ the interior vacuum of refrigerating chamber to preserve.
4. described a kind of suspension is inoculated the method for cultivating the summer truffle offspring according to claim 1, it is characterized in that: among the described inoculation step D, described inoculation culture medium uses 121-126 ℃ of high pressure 1.5kg/cm of high temperature
2The aseptic substrate of sterilization more than 2 hours, according to the difference of seeds, the peat soil that pinus yunnanensis should be adopted and vermiculite ratio by volume 1:1 are mixed; The naturally native and vermiculite ratio that Huashan pine and Chinese chestnut should be adopted by volume 1:1 is mixed; The pH value of matrix is regulated with quicklime, maintains 7.5.
5. the described a kind of suspension inoculation method of cultivating the summer truffle offspring according to claim 1 is characterized in that: described method of selecting the host plant seed is, pinus yunnanensis and Huashan pine are removed with wet concentration and dry rear normal temperature after the ghost and be in store for; The Castanea Mollissima Seeds wet concentration remove to go rotten and not full seed, dries surface moisture, again with humidity be 25-35% clean river sand by volume the ratio of 1:2 mix, and place 5 ℃ refrigerator to save backup.
6. described a kind of suspension is inoculated the method for cultivating the summer truffle offspring according to claim 1, it is characterized in that: among the step C of described artificial infection, and described inoculum concentration 2-5g/ strain.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103548575A (en) * | 2013-11-12 | 2014-02-05 | 中国科学院昆明植物研究所 | Tuber Magnatum mycorrhiza synthetic method |
| CN103733884A (en) * | 2014-01-13 | 2014-04-23 | 攀枝花市丰盛源农林开发有限公司 | Woodland microenvironment reform propagation promoting method of truffles |
| CN105475067A (en) * | 2015-11-28 | 2016-04-13 | 楚雄彝族自治州林业科学研究所 | Chinese chestnut tuber indicum mycorrhizal seedling culture method based on symbiotic relation |
| CN105474992A (en) * | 2015-11-28 | 2016-04-13 | 楚雄彝族自治州林业科学研究所 | Chinese chestnut summer truffle mycorrhizal seedling culture method based on symbiotic relation |
| CN106941930A (en) * | 2017-03-21 | 2017-07-14 | 会东县地金原松露产业发展有限公司 | A kind of cultural method of ferfas |
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| CN105474992A (en) * | 2015-11-28 | 2016-04-13 | 楚雄彝族自治州林业科学研究所 | Chinese chestnut summer truffle mycorrhizal seedling culture method based on symbiotic relation |
| CN106941930A (en) * | 2017-03-21 | 2017-07-14 | 会东县地金原松露产业发展有限公司 | A kind of cultural method of ferfas |
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