CN105039178A - Phlebopus portentosus basidiospore germination method - Google Patents
Phlebopus portentosus basidiospore germination method Download PDFInfo
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- CN105039178A CN105039178A CN201510495116.1A CN201510495116A CN105039178A CN 105039178 A CN105039178 A CN 105039178A CN 201510495116 A CN201510495116 A CN 201510495116A CN 105039178 A CN105039178 A CN 105039178A
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Abstract
The invention relates to a phlebopus portentosus basidiospore germination method, which belongs to the technical field of edible fungi breeding. The method has the following characteristics: a, prepared spore suspension liquid is placed in an incubator of 45 DEG C for culturing for 48 hours, and the spore suspension liquid is pretreated; b, a process of preparing a spore germination medium comprises the following steps: weighing red soil of 200g and peeled potato of 200g, adding water for boiling for 30 minutes, filtering with gauze, adding glucose of 20g, KH2PO4 of 1g, MgSO2 of 1g, yeast cream leach liquor of 2g and agar powder of 20g, adding water for a constant volume of 1000ml, enabling a pH value to be natural, sterilizing for 30 minutes at 121 DEG C, after a mixture is cooled to be below 60 DEG C and not solidified, adding penicillin and streptomycin to cause the final concentration to be 100U/ml, mixing and then pouring on a flat plate for standby. The method has the beneficial effects that time required for spore germination is greatly shortened, basidiospore begins germination by only needing 9-11 days, excellent foundation is laid for crossbreeding and genetic research of phlebopus portentosus in the future, and an application prospect is wide.
Description
Technical field
The present invention relates to a kind of method of Phlebopus portentosus basidiospore germination, belong to the Breeding of Edible Mushroom technical field.
Background technology
Phlebopus portentosus,
phlebopusportentosus(Berk & Broome) Boedijn, be under the jurisdiction of the little Boletaceae of Boletales (Boletinellaceae), mainly be distributed in general torrid areas, comprise the ground such as domestic Yunnan, Sichuan, Guangxi, Hainan, the ground such as external Thailand, Sri Lanka, Vietnam, Indonesia, Australia, New Zealand, Africa, Mexico.Phlebopus portentosus popular name " Boletus aereus ", its delicious flavour, nutritious, be one of most popular wild edible fungus of south of Yunnan and Northern Thailand, and be the edible bolete that uniquely really can realize the cultivation of artificial mushroom room known at present.When departing from host tree and being inoculated in artificial medium, Phlebopus portentosus sporidium, mycelium can be sprouted in laboratory conditions, grow, mating, through the cultivation of about 3 months in mushroom room, the whole life history can be completed, form sporophore.In recent years, about the correlative study of this bolete greenhouse production, field cultivation aspect continues to bring out, these researchs are the industrialization of this bolete, solid basis has been established in large-scale production, and its commercialization Extend culture is shown in blank.On the other hand, Phlebopus portentosus can realize within the shorter cycle, the circulation of sexual generation is completed under the condition of artificial greenhouse, this for the various environmental factorss in research its of process life history, obtain abundant filial generation and carry out genetics research, and the checking carrying out gene function is all very favourable condition, therefore one of its model animals being used as Boletales can be studied.
The single-ascospore strain obtained by spore germination, as the haploid stages in basidiomycetes life history, has and important effect in cross-breeding, basidiomycetes genetics research.Crossbreeding technology uses in edible mushrooms breeding of new variety the most extensively, and produce effects the most significant breeding technique, and its principle realizes gene recombination by monoploid mating, and from filial generation, seed selection has the bacterial strain of parents' good character concurrently.In the edible mushrooms of conventional cultivation, more than 80% of improved seeds are all derive from monospore cross-breeding.In addition, carrying out the drafting of basidiomycetes genetic map, Quantitative Trait Genes is located, and during the genetics research such as gene transformation, Monospore mycelium is also ideal material.
Summary of the invention
The object of the invention is to the deficiency overcoming prior art, and a kind of method of Phlebopus portentosus basidiospore germination is provided.
The inventive method is by improvement pretreatment process and spore germination substratum, 9-11 days its sporidium are only needed namely to start to sprout, substantially reduce spore germination required time, for the cross-breeding of Phlebopus portentosus in future, genetics research are had laid a good foundation, application prospect is extensive.
Phlebopus portentosus basidiospore germination method of the present invention, comprises the following steps:
(1) sporidium is collected: select Zhong Jun hole, artificial mushroom room just to start open Phlebopus portentosus sporophore, with the test tube of sterilizing in advance, aseptically select lower part cap, it is made to stick at test tube mouth place, test tube is placed in gnotobasis hold over night, sporidium is launched bottom test tube naturally, after 24 hours, sporophore block is taken out when naked eyes are shown on test tube wall and bottom occurs that brown sporidium prints, with spirit lamp flame, sterilising treatment is carried out to test tube mouth, get back into tampon, complete sporidium and collect;
(2) spore suspension pre-treatment: add 10ml sterilized water in above-mentioned containing in the test tube of Phlebopus portentosus sporidium, make spore suspension, is placed in 45 DEG C of incubators and cultivates 48 hours, carry out pre-treatment to spore suspension;
(3) spore germination is cultivated: aseptically dilute pretreated spore suspension with sterilized water, when having 10 ~ 20 sporidium to 100 powered microscope fields, coated Phlebopus portentosus basidiospore germination substratum, be placed in 28 DEG C of cultivations, 9-11 days basidiospore germinations;
Phlebopus portentosus basidiospore germination culture medium prescription and making method are: take 200 grams of laterite earth, and 200 grams of peeled potatoes add water boil after 30 minutes, filtered through gauze, add glucose 20 grams, KH
2pO
41 gram, MgSO
21 gram, yeast extract paste vat liquor 2 grams, agar powder 20 grams, adds water and is settled to 1000ml, pH value nature, 121 DEG C of sterilizings 30 minutes, when it is cooled to less than 60 DEG C but does not solidify, add penicillin and Streptomycin sulphate, make its final concentration be 100U/ml, be down flat plate after mixing immediately for subsequent use.
Beneficial effect of the present invention is: by improvement Phlebopus portentosus pretreatment process and spore germination substratum, substantially reduce spore germination required time, 9-11 days its sporidium are only needed namely to start to sprout, improve spore germination culture efficiency, for the cross-breeding of Phlebopus portentosus in future, genetics research are had laid a good foundation, application prospect is extensive.
Specific embodiment
The present invention's basidiospore germination method is sprouted Phlebopus portentosus sporidium, and concrete steps are as follows:
(1) sporidium is collected: select Zhong Jun hole, artificial mushroom room just to start open Phlebopus portentosus sporophore, utilize the test tube of sterilizing in advance, aseptically select lower part cap, it is made to stick at test tube mouth place, test tube is placed in gnotobasis hold over night, sporidium is launched bottom test tube naturally, after 24 hours, on the visible test tube wall of naked eyes and bottom occurs that brown sporidium prints, take out sporophore block, utilize spirit lamp flame to carry out sterilising treatment to test tube mouth, get back into tampon, complete sporidium and collect.
(2) spore suspension pre-treatment: add 10ml sterilized water in above-mentioned containing in the test tube of Phlebopus portentosus sporidium, make spore suspension, is placed in 45 DEG C of incubators and cultivates 48 hours, carry out pre-treatment to spore suspension;
(3) spore germination is cultivated: be aseptically diluted in 100 powered microscope fields by pretreated spore suspension and have 10 ~ 20 sporidium, coated Phlebopus portentosus basidiospore germination substratum, be placed in 28 DEG C of cultivations, 9 days ~ 11 days basidiospore germinations.
Phlebopus portentosus basidiospore germination culture medium prescription and making method are: take 200 grams of laterite earth, and 200 grams of peeled potatoes add water boil after 30 minutes, filtered through gauze, add glucose 20 grams, KH
2pO
41 gram, MgSO
21 gram, yeast extract paste vat liquor 2 grams, agar powder 20 grams, adds water and is settled to 1000ml, pH value nature, 121 DEG C of sterilizings 30 minutes, treat that it is cooled to less than 60 DEG C, but when not solidifying, add penicillin and Streptomycin sulphate, make its final concentration be 100U/ml, be down flat plate after mixing immediately for subsequent use.
Practical application shows: the inventive method spore germination is cultivated and only needed 9 days-11 days, and common PDA substratum spore germination is cultivated and then needed 29 days-31 days, and the inventive method substantially reduces spore germination required time, improves spore germination culture efficiency.
Claims (1)
1. a method for Phlebopus portentosus basidiospore germination, comprises sporidium and collects, spore suspension pre-treatment, and spore germination culturing step; It is characterized in that:
A., in spore suspension pre-treatment step: the spore suspension made, be placed in 45 DEG C of incubators and cultivate 48 hours, pre-treatment is carried out to spore suspension;
B. spore germination substratum is: take 200 grams of laterite earth, and 200 grams of peeled potatoes add water boil after 30 minutes, filtered through gauze, add glucose 20 grams, KH
2pO
41 gram, MgSO
21 gram, yeast extract paste vat liquor 2 grams, agar powder 20 grams, adds water and is settled to 1000ml, pH value nature, 121 DEG C of sterilizings 30 minutes, treat that it is cooled to less than 60 DEG C, when not solidifying, add penicillin and Streptomycin sulphate, make its final concentration be 100U/ml, be down flat plate after mixing immediately for subsequent use.
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| CN201510495116.1A CN105039178A (en) | 2015-08-13 | 2015-08-13 | Phlebopus portentosus basidiospore germination method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109097288A (en) * | 2018-08-02 | 2018-12-28 | 周茂 | A kind of fluid nutrient medium, the preparation method of Phlebopus portentosus batch production liquid spawn and the purposes of the fluid nutrient medium |
| CN111727806A (en) * | 2020-06-17 | 2020-10-02 | 云南省热带作物科学研究所 | Rapid germination method for phlebopus portentosus sclerotium |
Citations (4)
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| CN101491195A (en) * | 2009-03-05 | 2009-07-29 | 云南省热带作物科学研究所 | Phlebopus portentosus cultivation method |
| CN102476973A (en) * | 2011-11-10 | 2012-05-30 | 云南省热带作物科学研究所 | Phlebopus portentosus artificial cultivation casing soil material |
| CN103109647A (en) * | 2013-03-08 | 2013-05-22 | 北京农学院 | Method for collecting bolete spores |
| CN103113148A (en) * | 2013-03-08 | 2013-05-22 | 北京农学院 | Culture medium for germinating boletus spore |
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2015
- 2015-08-13 CN CN201510495116.1A patent/CN105039178A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101491195A (en) * | 2009-03-05 | 2009-07-29 | 云南省热带作物科学研究所 | Phlebopus portentosus cultivation method |
| CN102476973A (en) * | 2011-11-10 | 2012-05-30 | 云南省热带作物科学研究所 | Phlebopus portentosus artificial cultivation casing soil material |
| CN103109647A (en) * | 2013-03-08 | 2013-05-22 | 北京农学院 | Method for collecting bolete spores |
| CN103113148A (en) * | 2013-03-08 | 2013-05-22 | 北京农学院 | Culture medium for germinating boletus spore |
Non-Patent Citations (2)
| Title |
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| 何明霞等: "暗褐网柄牛肝菌菌丝生物学特性研究", 《云南省热带作物学会第七次代表大会暨2009年学术年会会议材料》 * |
| 吴圣进等: "热激及几个营养因子对草菇担孢子萌发的影响", 《广西农业科学》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109097288A (en) * | 2018-08-02 | 2018-12-28 | 周茂 | A kind of fluid nutrient medium, the preparation method of Phlebopus portentosus batch production liquid spawn and the purposes of the fluid nutrient medium |
| CN111727806A (en) * | 2020-06-17 | 2020-10-02 | 云南省热带作物科学研究所 | Rapid germination method for phlebopus portentosus sclerotium |
| CN111727806B (en) * | 2020-06-17 | 2022-03-18 | 云南省热带作物科学研究所 | Rapid germination method for phlebopus portentosus sclerotium |
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Application publication date: 20151111 |