CN105766382A - Antrodia cultivating method capable of improving content of triterpene - Google Patents
Antrodia cultivating method capable of improving content of triterpene Download PDFInfo
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- CN105766382A CN105766382A CN201610351283.3A CN201610351283A CN105766382A CN 105766382 A CN105766382 A CN 105766382A CN 201610351283 A CN201610351283 A CN 201610351283A CN 105766382 A CN105766382 A CN 105766382A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
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Abstract
The invention discloses an antrodia cultivating method.The method comprises the steps of strain culture medium manufacturing, strain isolation and expanding propagation, carrier cultivating, inoculation, mycelial growth and sporocarp growth.The antrodia cultivating method has the advantages that due to the cultivating method is simple and capable of achieving industrialization and standardized production, so that expanded production is promoted, the production cost is lowered, and the cultivating time is short.Low-density wood is adopted as a bacteria bed, mycelial growth is promoted, hyphae can easily form sporocarp, and meanwhile strains can better adapt to bacteria bed growth by improving the strains, so that the fruiting rate can be improved by about 80%, and economic benefits can be improved.The obtained antrodia sporocarp and wild antrodia sporocarp DNA detection equal rate can reach 99%, and the effect is good.The cultivating method can enable the content of triterpene of the antrodia sporocarp can reach 8.6%, the content of polysaccharide can reach 4.8%, and the content of effective elements can be improved.
Description
Technical field
The present invention relates to mushroom and cultivate field, particularly to a kind of Antrodia Camphorata breeding method improving triterpene content.
Background technology
Antrodia Camphorata has another name called Antrodia camphorata, it it is the medicinal fungi of a kind of preciousness being grown on Cinnamomum kanahirai hay tree, containing Multiple components such as polysaccharide body, terpenoid, sudismase and nucleic acids in Antrodia Camphorata, have removing toxic substances protect the liver, anticancer, reinforced immunological, antiallergic, the multiple efficacies such as blood fat reducing, be subject to the very big welcome of people.Antrodia Camphorata bacterium is relatively strong to the selectivity of substrate, is a kind of typical, domestomycetes that specificity is very strong.
The artificial cultivation method of current Antrodia Camphorata includes Cinnamomum kanahirai hay tree cultivation basswood method, solid-state cultivation and solution fermentation.Solid-state cultivation is that with space bag, Antrodia Camphorata strain is carried out Mycelium culture.Space includes fibre object, carbohydrate, dairy products etc.；Incubation time about three months；Toxigenic capacity is higher；The shortcoming of this breeding method is: triterpenes content is low, and effect is poor, is unfavorable for promoting.Solution fermentation be utilize 500 liters even tonne more than liquid fermentation groove carry out strain liquid and ferment to collect mycelium；Incubation time is short, is seven to fortnight；The distinctive triterpenes of wild Antrodia camphorata cannot be obtained.The shortcoming of this breeding method is: without triterpenes content, effect is poor, is unfavorable for promoting.Cinnamomum kanahirai hay tree cultivation basswood method is to utilize the original host Cinnamomum kanahirai hay tree of Antrodia camphorata or other basswoods (age of tree all basswood) more than 80 years to cultivate Antrodia Camphorata for culture medium；Obtaining the composition identical with wild Antrodia Camphorata, effect is identical, it is to avoid wild Cinnamomum kanahirai hay tree illegal logging；The shortcoming of this breeding method is: incubation time is long, is more than 3 years, and yield is relatively low, average Cinnamomum kanahirai hay per ton wood, within 3 years, just produces 10 kilograms of Antrodia Camphorata.
Summary of the invention
For solving the problem existing for the artificial cultivation method of above-mentioned tradition Antrodia Camphorata, it is desirable to provide a kind of nutrient content is high and cultivates the Antrodia Camphorata breeding method of low the improved triterpene content of cost.
For this, inventor provide following technical scheme:
A kind of Antrodia Camphorata breeding method improving triterpene content, it includes spawn culture matrix manufacturing, strain separating and expanding propagation, culture carrier, connects bacterium, mycelial growth, sporophore growth step.
Further, described spawn culture matrix manufacturing includes mother culture media and prepares, and the preparation of original seed or Cultivar culture medium.In the present invention, plant from mother and start to cultivate strain, it is ensured that the quality of strain and productivity；Meanwhile, gained Antrodia Camphorata sporophore rate identical with wild Antrodia Camphorata DNA of fruiting body detection reaches 99%, basically reaches effect of wild Antrodia Camphorata.
Further, the preparation of described mother culture media comprises the following steps:
1) component of following parts by weight is weighed:
Base material 150-250 part；
Glucose 10-30 part；
Agar 10-30 part；
Described base material is one of the following: Rhizoma Solani tuber osi, corn grit, hybridization rice or wheat grain；
In the present invention, mother culture media adopts above-mentioned component, nutritious, balanced, can improve the survival rate of female kind.
2) base material is added water boil 20-30min, filter, in filter cake, add agar, heat and stir, making agar all dissolve, then in solution, add glucose, add water after stirring and dissolving and solution is supplied to 1000 weight portions；
3) then by solution subpackage test tube, often pipe dress 1/4-1/3 test tube height, tampon beyond the Great Wall, at 110-130 DEG C of sterilizing 25-35min, to take out and be put into inclined-plane, cooled and solidified is stand-by.
Further, described step 1) in component also include potassium dihydrogen phosphate 2-5 weight portion, magnesium sulfate 1-4 weight portion and vitaminB10 .01-0.1 weight portion；Step 2) in filter cake, add agar while, also said components is all added in filter cake stirring and dissolving together.In the present invention, adding said components in mother culture media, be substantially shorter the incubation time of female kind, described potassium dihydrogen phosphate component can also promote the strongr of mycelial growth.
Further, the preparation method of described original seed or Cultivar culture medium comprises the following steps:
1) component of following parts by weight is weighed:
2) said components is mixed with water 1:1.3 in mass ratio stir evenly after bottling or pack, after sealing at 110 DEG C-130 DEG C sterilizing 1.5～2h, cool down stand-by.
Mother culture is become original seed or cultigen by pedigree seed culture medium or Cultivar culture medium by the present invention, thus realizing the amplifying incubation of female kind；It is diluted at the same time it can also be mother is planted, because according to pure female cultivation of planting, female consumption of planting is big, and cost is high, and it is long that mycelia sends out the bacterium time.
Further, the operation of described strain separating and expanding propagation comprises the following steps:
Obtaining Antrodia Camphorata, in inoculating hood, pelletizing loads in the test tube of mother culture media, cultivates three months, obtain Antrodia Camphorata mother's kind at 0 DEG C；Described Antrodia Camphorata granule weight 0.3-0.5g；Adopting this level of female kind to cultivate, regeneration capacity is strong, and survival rate can reach 90%-100%, is greatly improved female kind survival rate.
Antrodia Camphorata mother planting access equipped with in the culture bottle of pedigree seed culture medium, at 10-15 DEG C, lucifuge is cultivated 5-8 month, obtains Antrodia Camphorata original seed；The mother of a described test tube plants the bottled culture bottle having pedigree seed culture medium of access 3～5, and the bottled cultivation liquid measure of each cultivation is 20ml；
Being accessed by Antrodia Camphorata original seed equipped with in the culture bag of Cultivar culture medium, at 10-15 DEG C, lucifuge is cultivated 5-8 month, obtains Antrodia camphorata culturing strain；A described culture bottle original seed accesses 20～30 packed culture bag having Cultivar culture medium, and in each culture bag, the solid state cultivation culture medium of dress is about 500g.
In strain separating of the present invention and expanding propagation step, as cultivating the Antrodia Camphorata that mother plants, it is possible to be the Antrodia Camphorata of natural growth, it is also possible to be the Antrodia Camphorata cultivated through breeding method of the present invention.No matter it is the Antrodia Camphorata adopting natural growth, or the Antrodia Camphorata that the present invention cultivates cultivates mother's kind, adopts Antrodia Camphorata sporophore nutritional labeling (triterpene and the polysaccharide) content that breeding method of the present invention is cultivated close.
Further, described carrier is cultivated and is comprised the following steps:
1) select diameter of a cross-section of a tree trunk 1.3 meters above the ground 18-22cm and density lower than 0.4kg/m3Low-density timber, be processed into the plank block of 10*20*6cm；
2) the plank block processed is put into pond soak, change water every day, soak more than five days；
3) the plank block after soaking is put into steriliser, temperature be 110-130 DEG C, pressure be 0.1-0.2MPa when, sterilizing 5-8 hour；
4) after sterilizing, by packaged for plank enter 15*25*15cm specification bagging, and in bagging load 6-12g Cinnamomum kanahirai hay tree powder, put vinyl cover, reload in steriliser, temperature be 110-130 DEG C, pressure 0.1-0.2MPa when, sterilizing 5-8 hour again.
5) after second time sterilizing terminates, take out, cooling.
In the present invention, adopt low-density timber as the cultivation bacterium bed of Antrodia Camphorata, owing to low-density wood tissue has porous, be conducive to regulating the transport of moisture, promote Antrodia Camphorata sporophore growth.Utilize the breeding method of the present invention, improve Antrodia Camphorata sporophore Main Ingredients and Appearance content.The general triterpene content of Antrodia Camphorata sporophore that existing employing Cinnamomum kanahirai hay basswood produces is about 4%, polyoses content about 1.5%.Adopting the breeding method of the present invention, the triterpene content in Antrodia Camphorata sporophore reaches 8.6%, and polyoses content reaches 4.8%, and the main nutrient composition content in Antrodia Camphorata sporophore is greatly improved.
Meanwhile, the breeding method of the present invention is utilized, it is possible to shorten the growth and maturity phase of Antrodia Camphorata sporophore.Antrodia Camphorata sporophore cultivated by existing employing Cinnamomum kanahirai hay basswood generally to grow 3 years or 3 years half, and Antrodia camphorata sporophore is just ripe.And adopt the breeding method of the present invention, and only needing 22-26 month, Antrodia Camphorata sporophore can maturation.It addition, utilize the breeding method of the present invention, it is possible to saving the use of Cinnamomum kanahirai hay wood, bacterium bed material easily obtains, with low cost, effectively reduces cost.
Further, described in when connecing bacterium operation, Antrodia camphorata culturing strain is loaded in bagging, is seeded in plank block one end, the inoculum concentration of each plank block is 5-10g.In this operation, it is not necessary to hole on plank block, directly Antrodia camphorata culturing strain is loaded bagging.When vertically placing plank block, Antrodia camphorata culturing strain can be fallen bottom sack by nature, and plank block end thereof contacts, so that mycelia starts growth from plank one end.Adopt said method, not only can omit drilling operation (be usually at present and bore an aperture in plank block one end, be seeded in aperture by strain), and mycelia can large area deposition, improve cultivating rate.
Further, the condition of described mycelial growth is: be placed on layer frame by the plank block lucifuge after connecing bacterium, controls ambient temperature and is 20-28 DEG C, and humidity 65%-75% cultivates 90～130 days.
Further, described sporophore growth step concrete operations are: after de-bag fruiting, plank block is put into cultivation room, cover plank block with black cloth, keep room air circulation, and air humidity is 65%-75%, cultivate 20-24 month, until Antrodia Camphorata sporophore stops growing, then gather.
Compared with prior art, the Antrodia Camphorata breeding method of the improved triterpene content of the present invention has the advantage that
1, owing to breeding method is simple, it may be achieved industrialization, standardized production, thus facilitating expanding production, production cost is reduced.
2, due to the fact that selection low-density timber is as bacterium bed, it is simple to mycelial growth, mycelia is more readily formed sporophore, the improvement of strain simultaneously makes strain more adapt to the growth of this bacterium bed, cultivating rate is more than 78%, thus nearly 80 times of cultivating rate can be improved, increases economic efficiency.
3, adopting the breeding method of the present invention, using Cinnamomum kanahirai hay wood powder is nutrition, and gained Antrodia Camphorata sporophore rate identical with wild Antrodia Camphorata DNA of fruiting body detection reaches 99%, and effect is good.
4, in the breeding method of the present invention, adopting low-density timber is that bacterium bed is more beneficial for hyphal development, it is more beneficial for absorbing proteins, therefore the sporophore cultivated improves the Antrodia Camphorata sporophore Main Ingredients and Appearance content of cultivation, the general triterpene content of Antrodia Camphorata sporophore produced with Cinnamomum kanahirai hay basswood is about 4%, polysaccharide about 1.5%, the breeding method of the present invention can make Antrodia Camphorata sporophore triterpene content reach 8.6%, and polyoses content reaches 4.8%；
5, the breeding method of the present invention is adopted can to shorten the growth and maturity phase of Antrodia Camphorata sporophore.Antrodia Camphorata sporophore cultivated by existing employing Cinnamomum kanahirai hay basswood generally to grow 3 years or 3 years half, and Antrodia camphorata sporophore is just ripe.And adopt the breeding method of the present invention, and only needing 20-24 month, Antrodia Camphorata sporophore can maturation.
6, the breeding method of the present invention is utilized, it is not necessary to adopting Cinnamomum kanahirai hay wood is bacterium bed, it is possible to saving the use of Cinnamomum kanahirai hay wood, bacterium bed material easily obtains, with low cost, effectively reduces cost.
Detailed description of the invention
By describing the technology contents of the present invention, structural feature in detail, being realized purpose and effect, it is explained in detail below in conjunction with embodiment.
Embodiment 1
A kind of Antrodia Camphorata breeding method improving triterpene content, it comprises the following steps:
Step 10): spawn culture matrix manufacturing, it includes mother culture media and prepares, and the preparation of original seed or Cultivar culture medium, particularly as follows:
The preparation of described mother culture media comprises the following steps:
11) component of following parts by weight is weighed:
12) base material is added water boil 20min, filter, in filter cake, add agar, potassium dihydrogen phosphate, magnesium sulfate and vitamin B1, heat and stir, make solid all dissolve, then in solution, add glucose, add water after stirring and dissolving and solution is supplied to 1000 weight portions；
13) then by solution subpackage test tube, often pipe dress 1/4-1/3 test tube height, tampon beyond the Great Wall, at 110 DEG C of sterilizing 35min, to take out and be put into inclined-plane, cooled and solidified is stand-by.
The preparation method of described original seed or Cultivar culture medium comprises the following steps:
14) component of following parts by weight is weighed:
15) said components is mixed with water 1:1.3 in mass ratio stir evenly after bottling or pack, after sealing at 110 DEG C DEG C sterilizing 2h, cool down stand-by.
Step 20): strain separating and expanding propagation, specifically include following steps:
21) obtaining Antrodia Camphorata, in inoculating hood, pelletizing loads in the test tube of mother culture media, cultivates three months, obtain Antrodia Camphorata mother's kind at 0 DEG C；Described Antrodia Camphorata granule weight 0.3g；Adopting this level of female kind to cultivate, regeneration capacity is strong, and survival rate can reach 90%-100%, is greatly improved female kind survival rate.
22) Antrodia Camphorata mother planting access equipped with in the culture bottle of pedigree seed culture medium, at 10-15 DEG C, lucifuge is cultivated 8 months, obtains Antrodia Camphorata original seed；The female kind of a described test tube accesses the 5 bottled culture bottles having pedigree seed culture medium, and the bottled cultivation liquid measure of each cultivation is 20ml；
23) accessing Antrodia Camphorata original seed equipped with in the culture bag of Cultivar culture medium, at 10-15 DEG C, lucifuge is cultivated 5 months, obtains Antrodia camphorata culturing strain；A described culture bottle original seed accesses the 30 packed culture bag having Cultivar culture medium, and in each culture bag, the solid state cultivation culture medium of dress is about 500g.
Step 30): culture carrier, specifically include following steps:
31) select diameter of a cross-section of a tree trunk 1.3 meters above the ground 22cm and density lower than 0.4kg/m3Low-density timber, be processed into the plank block of 10*20*6cm；
32) the plank block processed is put into pond soak, change water every day, soak more than five days；
33) the plank block after soaking is put into steriliser, temperature be 130 DEG C, pressure be 0.1MPa when, sterilizing 5 hours；
34) after sterilizing, by packaged for plank enter 15*25*15cm specification bagging, and in bagging, load 6-12g Cinnamomum kanahirai hay tree powder, put vinyl cover, reload in steriliser, temperature be 130 DEG C, pressure 0.1MPa when, sterilizing 5 hours again.
35) after second time sterilizing terminates, take out, cooling.
Step 40): connecing bacterium, particularly as follows: loaded in bagging by Antrodia camphorata culturing strain, be seeded in plank block one end, the inoculum concentration of each plank block is 5g.
Step 50): mycelial growth, the condition of described mycelial growth is: be placed on layer frame by the plank block lucifuge after connecing bacterium, and controlling ambient temperature is 28 DEG C, and humidity 65% is cultivated 90 days.
Step 60): sporophore growth step, particularly as follows: after de-bag fruiting, plank block is put into cultivation room, cover plank block with black cloth, keep room air circulation, and air humidity is 75%, cultivate 20 months, until Antrodia Camphorata sporophore stops growing, then gather.
Embodiment 2
A kind of Antrodia Camphorata breeding method improving triterpene content, it comprises the following steps:
Step 10): spawn culture matrix manufacturing, it includes mother culture media and prepares, and the preparation of original seed or Cultivar culture medium, is specially
The preparation of described mother culture media comprises the following steps:
11) component of following parts by weight is weighed:
12) base material is added water boil 30min, filter, in filter cake, add agar, potassium dihydrogen phosphate, magnesium sulfate and vitamin B1, heat and stir, make solid all dissolve, then in solution, add glucose, add water after stirring and dissolving and solution is supplied to 1000 weight portions；
13) then by solution subpackage test tube, often pipe dress 1/4-1/3 test tube height, tampon beyond the Great Wall, at 130 DEG C of sterilizing 25min, to take out and be put into inclined-plane, cooled and solidified is stand-by.
The preparation method of described original seed or Cultivar culture medium comprises the following steps:
14) component of following parts by weight is weighed:
15) said components is mixed with water 1:1.3 in mass ratio stir evenly after bottling or pack, after sealing at 130 DEG C sterilizing 1.5h, cool down stand-by.
Step 20): strain separating and expanding propagation, specifically include following steps:
21) obtaining Antrodia Camphorata, in inoculating hood, pelletizing loads in the test tube of mother culture media, cultivates three months, obtain Antrodia Camphorata mother's kind at 0 DEG C；Described Antrodia Camphorata granule weight 0.5g；Adopting this level of female kind to cultivate, regeneration capacity is strong, and survival rate can reach 90%-100%, is greatly improved female kind survival rate.
22) Antrodia Camphorata mother planting access equipped with in the culture bottle of pedigree seed culture medium, at 10-15 DEG C, lucifuge is cultivated 8 months, obtains Antrodia Camphorata original seed；The female kind of a described test tube accesses the 3 bottled culture bottles having pedigree seed culture medium, and the bottled cultivation liquid measure of each cultivation is 20ml；
23) accessing Antrodia Camphorata original seed equipped with in the culture bag of Cultivar culture medium, at 10-15 DEG C, lucifuge is cultivated 5 months, obtains Antrodia camphorata culturing strain；A described culture bottle original seed accesses the 20 packed culture bag having Cultivar culture medium, and in each culture bag, the solid state cultivation culture medium of dress is about 500g.
Step 30): culture carrier, specifically include following steps:
31) select diameter of a cross-section of a tree trunk 1.3 meters above the ground 18cm and density lower than 0.4kg/m3Low-density timber, be processed into the plank block of 10*20*6cm；
32) the plank block processed is put into pond soak, change water every day, soak more than five days；
33) the plank block after soaking is put into steriliser, temperature be 110 DEG C, pressure be 0.2MPa when, sterilizing 8 hours；
34) after sterilizing, by packaged for plank enter 15*25*15cm specification bagging, and in bagging, load 6g Cinnamomum kanahirai hay tree powder, put vinyl cover, reload in steriliser, temperature be 110 DEG C, pressure 0.2MPa when, sterilizing 8 hours again.
35) after second time sterilizing terminates, take out, cooling.
Step 40): connecing bacterium, particularly as follows: loaded in bagging by Antrodia camphorata culturing strain, be seeded in plank block one end, the inoculum concentration of each plank block is 10g.
Step 50): mycelial growth, the condition of described mycelial growth is: be placed on layer frame by the plank block lucifuge after connecing bacterium, and controlling ambient temperature is 20 DEG C, and humidity 75% is cultivated 130 days.
Step 60) sporophore growth step, particularly as follows: after de-bag fruiting, plank block is put into cultivation room, cover plank block with black cloth, keep room air circulation, and air humidity is 65%, cultivate 24 months, until Antrodia Camphorata sporophore stops growing, then gather.
Embodiment 3
A kind of Antrodia Camphorata breeding method improving triterpene content, it comprises the following steps:
Step 10): spawn culture matrix manufacturing, it includes mother culture media and prepares, and the preparation of original seed or Cultivar culture medium, is specially
The preparation of described mother culture media comprises the following steps:
11) component of following parts by weight is weighed:
12) base material is added water boil 25min, filter, in filter cake, add agar, potassium dihydrogen phosphate, magnesium sulfate and vitamin B1, heat and stir, make solid all dissolve, then in solution, add glucose, add water after stirring and dissolving and solution is supplied to 1000 weight portions；
13) then by solution subpackage test tube, often pipe dress 1/4-1/3 test tube height, tampon beyond the Great Wall, at 120 DEG C of sterilizing 30min, to take out and be put into inclined-plane, cooled and solidified is stand-by.
The preparation method of described original seed or Cultivar culture medium comprises the following steps:
14) component of following parts by weight is weighed:
15) said components is mixed with water 1:1.3 in mass ratio stir evenly after bottling or pack, after sealing at 120 DEG C sterilizing 1.5h, cool down stand-by.
Step 20): strain separating and expanding propagation, specifically include following steps:
21) obtaining Antrodia Camphorata, in inoculating hood, pelletizing loads in the test tube of mother culture media, cultivates three months, obtain Antrodia Camphorata mother's kind at 0 DEG C；Described Antrodia Camphorata granule weight 4g；Adopting this level of female kind to cultivate, regeneration capacity is strong, and survival rate can reach 90%-100%, is greatly improved female kind survival rate.
22) Antrodia Camphorata mother planting access equipped with in the culture bottle of pedigree seed culture medium, at 10-15 DEG C, lucifuge is cultivated 6 months, obtains Antrodia Camphorata original seed；The mother of a described test tube plants the bottled culture bottle having pedigree seed culture medium of access 3～5, and the bottled cultivation liquid measure of each cultivation is 20ml；
23) accessing Antrodia Camphorata original seed equipped with in the culture bag of Cultivar culture medium, at 10-15 DEG C, lucifuge is cultivated 7 months, obtains Antrodia camphorata culturing strain；A described culture bottle original seed accesses the 25 packed culture bag having Cultivar culture medium, and in each culture bag, the solid state cultivation culture medium of dress is about 500g.
Step 30): culture carrier, specifically include following steps:
31) select diameter of a cross-section of a tree trunk 1.3 meters above the ground 20cm and density lower than 0.4kg/m3Low-density timber, be processed into the plank block of 10*20*6cm；
32) the plank block processed is put into pond soak, change water every day, soak more than five days；
33) the plank block after soaking is put into steriliser, temperature be 120 DEG C, pressure be 0.15MPa when, sterilizing 6 hours；
34) after sterilizing, by packaged for plank enter 15*25*15cm specification bagging, and in bagging, load 8g Cinnamomum kanahirai hay tree powder, put vinyl cover, reload in steriliser, temperature be 120 DEG C, pressure 0.15MPa when, sterilizing 6 hours again.
35) after second time sterilizing terminates, take out, cooling.
Step 40): connecing bacterium, particularly as follows: loaded in bagging by Antrodia camphorata culturing strain, be seeded in plank block one end, the inoculum concentration of each plank block is 7g.
Step 50): mycelial growth, the condition of described mycelial growth is: be placed on layer frame by the plank block lucifuge after connecing bacterium, and controlling ambient temperature is 22 DEG C, and humidity 70% is cultivated 120 days.
Step 60): sporophore growth step, particularly as follows: after de-bag fruiting, plank block is put into cultivation room, cover plank block with black cloth, keep room air circulation, and air humidity is 70%, cultivate 21 months, until Antrodia Camphorata sporophore stops growing, then gather.
Embodiment 4
A kind of Antrodia Camphorata breeding method improving triterpene content, it comprises the following steps:
Step 10): spawn culture matrix manufacturing, it includes mother culture media and prepares, and the preparation of original seed or Cultivar culture medium, is specially
The preparation of described mother culture media comprises the following steps:
11) component of following parts by weight is weighed:
12) base material is added water boil 25min, filter, in filter cake, add agar, potassium dihydrogen phosphate, magnesium sulfate and vitamin B1, heat and stir, make solid all dissolve, then in solution, add glucose, add water after stirring and dissolving and solution is supplied to 1000 weight portions；
13) then by solution subpackage test tube, often pipe dress 1/4-1/3 test tube height, tampon beyond the Great Wall, at 125 DEG C of sterilizing 33min, to take out and be put into inclined-plane, cooled and solidified is stand-by.
The preparation method of described original seed or Cultivar culture medium comprises the following steps:
14) component of following parts by weight is weighed:
15) said components is mixed with water 1:1.3 in mass ratio stir evenly after bottling or pack, after sealing at 115 DEG C sterilizing 1.8h, cool down stand-by.
Step 20): strain separating and expanding propagation, specifically include following steps:
21) obtaining Antrodia Camphorata, in inoculating hood, pelletizing loads in the test tube of mother culture media, cultivates three months, obtain Antrodia Camphorata mother's kind at 0 DEG C；Described Antrodia Camphorata granule weight 0.5g；Adopting this level of female kind to cultivate, regeneration capacity is strong, and survival rate can reach 90%-100%, is greatly improved female kind survival rate.
22) Antrodia Camphorata mother planting access equipped with in the culture bottle of pedigree seed culture medium, at 10-15 DEG C, lucifuge is cultivated 5 months, obtains Antrodia Camphorata original seed；The female kind of a described test tube accesses the 4 bottled culture bottles having pedigree seed culture medium, and the bottled cultivation liquid measure of each cultivation is 20ml；
23) accessing Antrodia Camphorata original seed equipped with in the culture bag of Cultivar culture medium, at 10-15 DEG C, lucifuge is cultivated 6 months, obtains Antrodia camphorata culturing strain；A described culture bottle original seed accesses the 25 packed culture bag having Cultivar culture medium, and in each culture bag, the solid state cultivation culture medium of dress is about 500g.
Step 30): culture carrier, specifically include following steps:
31) select diameter of a cross-section of a tree trunk 1.3 meters above the ground 21cm and density lower than 0.4kg/m3Low-density timber, be processed into the plank block of 10*20*6cm；
32) the plank block processed is put into pond soak, change water every day, soak more than five days；
33) the plank block after soaking is put into steriliser, temperature be 130 DEG C, pressure be 0.1MPa when, sterilizing 5 hours；
34) after sterilizing, by packaged for plank enter 15*25*15cm specification bagging, and in bagging, load 10g Cinnamomum kanahirai hay tree powder, put vinyl cover, reload in steriliser, temperature be 130 DEG C, pressure 0.1MPa when, sterilizing 5 hours again.
35) after second time sterilizing terminates, take out, cooling.
Step 40): connecing bacterium, particularly as follows: loaded in bagging by Antrodia camphorata culturing strain, be seeded in plank block one end, the inoculum concentration of each plank block is 8g.
Step 50): mycelial growth, the condition of described mycelial growth is: be placed on layer frame by the plank block lucifuge after connecing bacterium, and controlling ambient temperature is 26 DEG C, and humidity 72% is cultivated 110 days.
Step 60): sporophore growth step, particularly as follows: after de-bag fruiting, plank block is put into cultivation room, cover plank block with black cloth, keep room air circulation, and air humidity is 70%, cultivate 22 months, until Antrodia Camphorata sporophore stops growing, then gather.
Cultivation time and cultivating rate to embodiment 1-embodiment 4 are added up, and compare with traditional Cinnamomum kanahirai hay tree cultivation basswood method, and result is in Table one.In traditional method, with Cinnamomum kanahirai hay wood for bacterium bed, the Cinnamomum kanahirai hay wood fruiting about 10 kilograms of 1 ton, its cultivating rate is 1%, and in the breeding method of the present invention, only need to need to add Cinnamomum kanahirai hay wood powder in carrier incubation step, and addition is few.So, calculate by cultivating base material Cinnamomum kanahirai hay wood weight, adopt breeding method cultivating rate of the present invention up to more than 78%.
Table 1 embodiment 1 cultivates time and the cultivating rate contrast of Antrodia Camphorata sporophore to embodiment 4 and prior art
As can be seen from Table 1, adopting the breeding method of the present invention, the cultivation time of Antrodia Camphorata sporophore is greatly shortened.Antrodia Camphorata sporophore cultivated by existing employing Cinnamomum kanahirai hay basswood generally to grow 3 years or 3 years half, and Antrodia camphorata sporophore is just ripe.And adopt the breeding method of the present invention, and only needing 23-26 month, Antrodia Camphorata sporophore can maturation.
By table 1 it can also be seen that adopt the breeding method of the present invention, cultivating rate about 80 times can be improved, increase economic efficiency.
Through DNA detection, the Antrodia Camphorata sporophore rate identical with wild Antrodia Camphorata DNA of fruiting body that embodiment of the present invention 1-4 cultivates reaches 99%, substantially can reach effect of wild Antrodia Camphorata.
Detecting through microbiological analysis inspection center of Guangdong Province, the Antrodia Camphorata sporophore that the content of the Antrodia Camphorata sporophore nutritional labeling (triterpene, polysaccharide) that embodiment 1-embodiment 4 is cultivated and prior art are cultivated contrasts in Table two.
Table 2 embodiment 1 cultivates each nutrient composition content contrast of Antrodia Camphorata sporophore to embodiment 4 and prior art
As shown in Table 2, the breeding method of the present invention improves the Antrodia Camphorata sporophore Main Ingredients and Appearance content of cultivation, and the general triterpene content of Antrodia Camphorata sporophore produced with Cinnamomum kanahirai hay basswood is about 4%, polysaccharide about 1.5%.The breeding method of the present invention can make sporophore triterpene content reach 8.6%, and polyoses content reaches 4.8%.Meanwhile, the present invention is as cultivating the Antrodia Camphorata that mother plants, and being possible not only to is the Antrodia Camphorata of natural growth, it is also possible to being the Antrodia Camphorata cultivated of breeding method of the present invention, triterpene and polyoses content all can reach above-mentioned detection level.
The breeding method of the present invention is simple, it may be achieved industrialization, standardized production, thus facilitating expanding production, reduces production cost.
The breeding method of the present invention is without adopting Cinnamomum kanahirai hay tree for bacterium bed, it is possible to saving the use of Cinnamomum kanahirai hay wood, bacterium bed material easily obtains, with low cost, effectively reduces cost.
The foregoing is only embodiments of the invention; not thereby the scope of patent protection of the present invention is limited; every equivalent structure utilizing description of the present invention to make or equivalence flow process conversion; or directly or indirectly it is used in other relevant technical fields, all in like manner include in the scope of patent protection of the present invention.
Claims (10)
1. the Antrodia Camphorata breeding method that can improve triterpene content, it is characterised in that: it includes spawn culture matrix manufacturing, strain separating and expanding propagation, culture carrier, connects bacterium, mycelial growth, sporophore growth step.
2. the Antrodia Camphorata breeding method improving triterpene content according to claim 1, it is characterised in that: described spawn culture matrix manufacturing includes mother culture media and prepares, and the preparation of original seed or Cultivar culture medium.
3. the Antrodia Camphorata breeding method improving triterpene content according to claim 2, it is characterised in that: the preparation of described mother culture media comprises the following steps:
1) component of following parts by weight is weighed:
Base material 150-250 part；
Glucose 10-30 part；
Agar 10-30 part；
Described base material is one of the following: Rhizoma Solani tuber osi, corn grit, hybridization rice or wheat grain；
2) base material is added water boil 20-30min, filter, in filter cake, add agar, heat and stir, making agar all dissolve, then in solution, add glucose, add water after stirring and dissolving and solution is supplied to 1000 weight portions；
3) then by solution subpackage test tube, often pipe dress 1/4-1/3 test tube height, tampon beyond the Great Wall, at 110-130 DEG C of sterilizing 25-35min, to take out and be put into inclined-plane, cooled and solidified is stand-by.
4. the Antrodia Camphorata breeding method improving triterpene content according to claim 3, it is characterised in that: described step 1) in component also include potassium dihydrogen phosphate 2-5 weight portion, magnesium sulfate 1-4 weight portion and vitaminB10 .01-0.1 weight portion；Step 2) in filter cake, add agar while, also said components is all added in filter cake stirring and dissolving together.
5. the Antrodia Camphorata breeding method improving triterpene content according to claim 2, it is characterised in that: the preparation method of described original seed or Cultivar culture medium comprises the following steps:
1) component of following parts by weight is weighed:
2) said components is mixed with water 1:1.3 in mass ratio stir evenly after bottling or pack, after sealing at 110 DEG C-130 DEG C sterilizing 1.5～2h, cool down stand-by.
6. the Antrodia Camphorata breeding method improving triterpene content according to claim 2, it is characterised in that: the operation of described strain separating and expanding propagation comprises the following steps:
Obtaining Antrodia Camphorata, in inoculating hood, pelletizing loads in the test tube of mother culture media, cultivates three months, obtain Antrodia Camphorata mother's kind at 0 DEG C；Described Antrodia Camphorata granule weight 0.3-0.5g；
Antrodia Camphorata mother planting access equipped with in the culture bottle of pedigree seed culture medium, at 10-15 DEG C, lucifuge is cultivated 5-8 month, obtains Antrodia Camphorata original seed；The mother of a described test tube plants the bottled culture bottle having pedigree seed culture medium of access 3～5；
Being accessed by Antrodia Camphorata original seed equipped with in the culture bag of Cultivar culture medium, at 10-15 DEG C, lucifuge is cultivated 5-8 month, obtains Antrodia camphorata culturing strain；A described culture bottle original seed accesses 20～30 packed culture bag having Cultivar culture medium.
7. the Antrodia Camphorata breeding method improving triterpene content according to claim 1, it is characterised in that: described carrier is cultivated and is comprised the following steps:
1) select diameter of a cross-section of a tree trunk 1.3 meters above the ground 18-22cm and density lower than 0.4kg/m3Low-density timber, be processed into the plank block of 10*20*6cm；
2) the plank block processed is put into pond soak, change water every day, soak more than five days；
3) the plank block after soaking is put into steriliser, temperature be 110-130 DEG C, pressure be 0.1-0.2MPa when, sterilizing 5-8 hour；
4) after sterilizing, by packaged for plank enter 15*25*15cm specification bagging, and in bagging load 6-12g Cinnamomum kanahirai hay tree powder, put vinyl cover, reload in steriliser, temperature be 110-130 DEG C, pressure 0.1-0.2MPa when, sterilizing 5-8 hour again.
5) after second time sterilizing terminates, take out, cooling.
8. the Antrodia Camphorata breeding method of the improved triterpene content according to claim 6 or 7, it is characterised in that: described in when connecing bacterium operation, Antrodia camphorata culturing strain is loaded in bagging, is seeded in plank block one end, the inoculum concentration of each plank block is 5-10g.
9. the Antrodia Camphorata breeding method of the improved triterpene content according to claim 6 or 7, it is characterized in that: the condition of described mycelial growth is: the plank block lucifuge after connecing bacterium is placed on layer frame, control ambient temperature and be 20-28 DEG C, humidity 65%-75%, cultivates 90～130 days.
10. the Antrodia Camphorata breeding method of the improved triterpene content according to claim 6 or 7, it is characterized in that: described sporophore growth step concrete operations are: after de-bag fruiting, plank block is put into cultivation room, plank block is covered with black cloth, keep room air circulation, and air humidity is 65%-75%, cultivate 20-24 month, until Antrodia Camphorata sporophore stops growing, then gather.
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| CN107409748A (en) * | 2017-05-17 | 2017-12-01 | 左春萌 | It is a kind of can factorial praluction Antrodia camphorata breeding method |
| CN109122020A (en) * | 2018-08-20 | 2019-01-04 | 江西省利财食用菌有限公司 | A kind of cultural method of antrodia |
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