CN103718961B - The sponge liquid medium that Antrodia camphorata cuttage branch survival rate is high and application thereof - Google Patents
The sponge liquid medium that Antrodia camphorata cuttage branch survival rate is high and application thereof Download PDFInfo
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Abstract
The invention discloses the sponge liquid medium that Antrodia camphorata cuttage branch survival rate is high, that on sponge, evenly have 15 bores be 0.8cm, the degree of depth is the V-shape hole of 0.5cm size, sponge block is lain in bottom blake bottle, one of perforate faces up, direct running water prepare medicines and through constant volume mix up pH value liquid nutrient medium, be dispensed in right amount and be placed with in the blake bottle of sponge, nutrient solution has just been invaded and finishes sponge block; The component of liquid nutrient medium consists of PDB medium+MY medium.Beneficial effect of the present invention is: sponge liquid medium provided by the invention, has the following advantages: 1, Antrodia camphorata group training material fully can absorb nutrition, improves quality and output, and the rate of increase can reach 5-6 doubly; 2, sponge can reuse, and liquid nutrient medium packing preparation is more convenient, and the apparatus such as bottle easily clean up; 3, save the expense buying agar powder, reduce production cost.
Description
Technical field
The present invention relates to the fast numerous field of plant tissue culture seedling, be specifically related to the high sponge liquid medium of Antrodia camphorata cuttage branch survival rate and application thereof.
Background technology
Camphor tree sesame has another name called Cinnamomum kanahirai hay mushroom, and belong to Aphyllophorales, porous section, perennial gill fungus mushroom, formal name used at school is (Antrodiacamphorata), is the novel species just delivered by biochemical boundary nineteen ninety.Its growth district is between the height above sea level 450-2000 meter mountain forest of mountain area, Taiwan, and the heartwood inwall that only the Cinnamomum kanahirai hay tree trunk of growth more than the peculiar a century in Taiwan is rotten, or the Cinnamomum kanahirai hay trees material moist surface of withered lodging, it gathers very not easily.At present, the survival rate of general Antrodia camphorata cuttage branch is 20%.
Summary of the invention
Object of the present invention is exactly for above-mentioned defect of the prior art, provides a kind of sponge liquid medium improving Antrodia camphorata cuttage branch survival rate.
To achieve these goals, technical scheme provided by the invention is: the sponge liquid medium that Antrodia camphorata cuttage branch survival rate is high, select columniform high temperature resistant filtration sponge, in the same size bottom diameter and ventilating cover thread bottle, sponge thickness is 1cm, on sponge, evenly have 15 bores is 0.8cm, the degree of depth is the V-shape hole of 0.5cm size, sponge block is lain in bottom blake bottle, one of perforate faces up, direct running water prepare medicines and through constant volume mix up pH value liquid nutrient medium, be dispensed in right amount and be placed with in the blake bottle of sponge, nutrient solution is just invaded and finishes sponge block, the component of described liquid nutrient medium consists of B5 medium+TDZ plant growth regulator 0.8mg/L+ methyl α-naphthyl acetate 0.2-0.8mg/L.
Second object of the present invention there is provided the application of above-mentioned sponge liquid medium in Antrodia camphorata cuttage branch is cultivated.
Further, above-mentioned application, comprises the following steps:
1) by the above-mentioned point sponge liquid medium installed through autoclaving, sterilising temp is 121-123 DEG C, and sterilization pressure is 0.11-0.13MP, and sterilization time is 22-25 minute, obtains sterilizing wild Oryza species;
2) inoculation of culture materials: under germ-free condition, Antrodia camphorata culture materials is cut into simple bud and insert respectively in the V-shape hole of planting the sponge liquid medium after sterilizing, carry out fast breeding cultivation, incubation time is 30-35 days, and the rate of increase is 5-6 times;
3) switching of propagation material: by step 2) in material after propagation hold under the arm out to cut to inoculate in fresh liquid nutrient medium and proceed Multiplying culture; After this, when cultivating 30-35 days, again material being carried out cutting changing in fresh liquid nutrient medium and carrying out Multiplying culture;
4) culture of rootage: after the Antrodia camphorata propagation material after Multiplying culture step 3) obtained is transferred to some, cut into individual plant to be transferred in the solid culture medium being added with agar powder and to carry out culture of rootage, after 2 ~ 3 months cultivate hardening, namely obtain Antrodia camphorata plantlet in vitro transplant strain.
Beneficial effect of the present invention is: sponge liquid medium provided by the invention, has the following advantages:
1, Antrodia camphorata group training material fully can absorb nutrition, improves quality and output, and the rate of increase can reach 5-6 doubly, and survival rate can reach 80%;
2, sponge can reuse, and liquid nutrient medium packing preparation is more convenient, and the apparatus such as bottle easily clean up;
3, save the expense buying agar powder, reduce production cost.
Embodiment
embodiment 1:
(1) making of sponge liquid medium:
Select columniform high temperature resistant filtration sponge, in the same size bottom diameter and ventilating cover thread bottle, sponge thickness is 1cm, on sponge, evenly have 15 bores is 0.8cm, the degree of depth is the V-shape hole of 0.5cm size, is lain in by sponge block bottom blake bottle, and one of perforate faces up.Because not adding agar powder, can direct running water prepare medicines and through constant volume mix up pH value liquid nutrient medium, be finally dispensed in right amount and be placed with in the blake bottle of sponge, nutrient solution has just been invaded and finishes sponge block;
(2) sterilizing of medium:
Because the sponge selected is exotic material, main component is polyurethane, have anticorrosive, have no side effect, high temperature resistant (can reach 200 DEG C), intensity high (resilience is good), water imbibition be extremely strong, the advantages such as recoverable, after repeatedly autoclaving, modification is still not reusable; Divide the medium installed through autoclaving (sterilising temp is 121-123 degree, and pressure is 0.11-0.13MP, and the time is 22-25 minute), finally the liquid nutrient medium that sterilizing is good is put into medium storeroom for subsequent use;
(3) inoculation of culture materials:
Culture materials cuts into simple bud by aseptic working platform and inserts kind respectively in the V-shape hole of sponge liquid medium, because sponge has good resilience and water imbibition, there are certain fixation and moisture-keeping functions to the culture materials be inserted in V-shape hole, are beneficial to the growth of culture and the absorption of nutrient;
(4) Multiplying culture of culture materials:
Cultivate the material on sponge liquid medium, due to liquid nutrient medium Middle molecule or ion motion very fast, its base portion fully can touch nutrient solution and can absorb nutrient preferably, also the quinones harmful substance that self otch is secreted is diluted in liquid nutrient medium simultaneously, reduce the generation of brownization, thus reach the effect of fast breeding, generally cultivate the rate of increase after 30-35 days and can reach 5-6 doubly;
(5) switching of propagation material:
After cultivating 30-35 days in propagation material liquid medium within, culture materials is proliferate constantly, has absorbed most nutrition and moisture, also have accumulated the harmful components such as certain quinones in liquid nutrient medium simultaneously; Now the material after propagation should be held under the arm out to cut to inoculate in fresh liquid nutrient medium and proceed Multiplying culture, after this when cultivating 30-35 days, constantly material is carried out cutting changing in fresh liquid nutrient medium and carry out Multiplying culture, production can be made to reach the material of some and scale;
(6) culture of rootage:
After Antrodia camphorata propagation material is transferred to the quantity needed for production, is cut into individual plant and be transferred in the solid culture medium being added with agar powder and carry out culture of rootage, can be transplanted by bottle outlet after 2 ~ 3 months cultivate hardening.
Last it is noted that the foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, although with reference to previous embodiment to invention has been detailed description, for a person skilled in the art, it still can be modified to the technical scheme described in foregoing embodiments, or carries out equivalent replacement to wherein portion of techniques feature.Within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (2)
1. the sponge liquid medium that Antrodia camphorata cuttage branch survival rate is high, it is characterized in that, select columniform high temperature resistant filtration sponge, in the same size bottom diameter and ventilating cover thread bottle, sponge thickness is 1cm, on sponge, evenly have 15 bores is 0.8cm, the degree of depth is the V-shape hole of 0.5cm size, sponge block is lain in bottom blake bottle, one of perforate faces up, direct running water prepare medicines and through constant volume mix up pH value liquid nutrient medium, be dispensed in right amount and be placed with in the blake bottle of sponge, nutrient solution has just been invaded and finishes sponge block; The component of described liquid nutrient medium consists of B5 medium+TDZ plant growth regulator 0.8mg/L+ methyl α-naphthyl acetate 0.2-0.8mg/L.
2. the application of the sponge liquid medium that Antrodia camphorata cuttage branch survival rate as claimed in claim 1 is high, is characterized in that, comprise the following steps:
1) will the sponge liquid medium installed be divided through autoclaving, sterilising temp be 121-123 DEG C, and sterilization pressure is 0.11-0.13MP, and sterilization time is 22-25 minute, obtains sterilizing wild Oryza species;
2) inoculation of culture materials: under germ-free condition, Antrodia camphorata culture materials is cut into simple bud and insert respectively in the V-shape hole of planting the sponge liquid medium after sterilizing, carry out fast breeding cultivation, incubation time is 30-35 days, and the rate of increase is 5-6 times;
3) switching of propagation material: by step 2) in material after propagation hold under the arm out to cut to inoculate in fresh liquid nutrient medium and proceed Multiplying culture; After this, when cultivating 30-35 days, again material being carried out cutting changing in fresh liquid nutrient medium and carrying out Multiplying culture;
4) culture of rootage: after the Antrodia camphorata propagation material after Multiplying culture step 3) obtained is transferred to some, cut into individual plant to be transferred in the solid culture medium being added with agar powder and to carry out culture of rootage, after 2 ~ 3 months cultivate hardening, namely obtain Antrodia camphorata plantlet in vitro transplant strain.
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| CN103734008B (en) * | 2013-12-26 | 2016-04-13 | 广西森仙源生物科技有限公司 | A kind of butterfly orchid tissue cultivating Fast-propagation liquid nutrient medium and corresponding cultural method |
| CN105766512A (en) * | 2015-11-19 | 2016-07-20 | 福建省亚热带植物研究所 | Cuttage method for Cinnamomum kanehirai |
| CN106105777A (en) * | 2016-07-01 | 2016-11-16 | 刘刚 | A kind of Flammulina velutiper (Fr.) Sing soilless culture method |
| CN106359067A (en) * | 2016-08-19 | 2017-02-01 | 明光现代农业科技合作推广服务中心 | Water-retaining and automatic water circulation cuttage device |
| CN108886988A (en) * | 2018-06-28 | 2018-11-27 | 普定县祝春农产品有限公司 | A kind of blueberry sponge cuttage and seedling culture method |
| CN111802170A (en) * | 2019-04-10 | 2020-10-23 | 四季洋圃生物机电股份有限公司 | Environment-friendly liquid cultivation method of mushroom |
| CN111264218A (en) * | 2020-04-07 | 2020-06-12 | 肇庆格伦夫利农业科学研究和试验发展有限公司 | Soilless cutting method for oasis I |
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| TWI377905B (en) * | 2009-12-02 | 2012-12-01 | Endemic Species Res Inst C O A | Method for incubating fruiting bodies of antrodia cinnamomea |
| CN102746054B (en) * | 2011-04-18 | 2014-05-07 | 甘泉生物科技有限公司 | Culture medium for culturing fruiting bodies of Antrodia cinnamomea and cultural method thereof |
| CN103155807B (en) * | 2011-12-13 | 2014-04-02 | 西北农林科技大学 | Plant callus grafting method and device thereof |
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