CN104957045A - Rapid induction medium for tissue culture of Alhagi sparsifolia Shap. and preparation method of rapid induction medium - Google Patents
Rapid induction medium for tissue culture of Alhagi sparsifolia Shap. and preparation method of rapid induction medium Download PDFInfo
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- CN104957045A CN104957045A CN201510462655.5A CN201510462655A CN104957045A CN 104957045 A CN104957045 A CN 104957045A CN 201510462655 A CN201510462655 A CN 201510462655A CN 104957045 A CN104957045 A CN 104957045A
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Abstract
The invention belongs to the technical field of plant culturing, and in particular relates to a rapid induction medium for tissue culture of Alhagi sparsifolia Shap. and a preparation method of the rapid induction medium. The rapid induction medium is prepared by adding a specially-made curing agent, brown sugar, 6-benzylaminopurine, 4-chlorine-IAA, tetracycline, active carbon, a plant extract and papaya juice into a 1/2MS culture medium. According to the rapid induction medium, tetracycline is added, so that obvious inhibition effect can be achieved on endophyte of an explant, meanwhile, an extract of aerial roots of ficus virens is matched with 4-chlorine-IAA to effectively balance growth of axillary buds, roots and stems of Alhagi sparsifolia Shap. and to play the obvious role of promoting germination of the axillary buds and development of the buds. When the induction medium provided by the invention is used to culture Alhagi sparsifolia Shap., the survival rate is 98%-99%, the infection rate is 1.7%-2.1%, the browning degree of the medium is 14%-17% lower than that of the MS culture medium, and the germinating time is shortened by 25%-35%.
Description
Technical field
The invention belongs to technical field of plant culture, be specifically related to rapid induction medium of a kind of camel thorn tissue cultures and preparation method thereof.
Background technology
Camel thorn (formal name used at school: Alhagi sparsifolia Shap.) belongs to pulse family, fallen leaves draft, thorniness on main branch, leaf Long Circle, and pollen is red, June blooms, and August contains most, and every flower can open more than 20 skies, bear pods really, raceme, root system generally reaches 20 meters.From desert and depths, Gobi desert groundwater absorbing part and nutrition, it is a kind of drought-enduring plant of self-sow, because this axis is had the very hard little greenery of perverse shape, therefore make camel perverse, it is herbaceous plant, be the grass of depending on for existence that in Gobi desert and desert, camel uniquely can eat, therefore have another name called camel grass, be mainly distributed in arid area, inland.
Camel thorn root system is very flourishing, and root system generally reaches 20 meters, and be 2 times even 3 times of cauline leaf radius on earth's surface, this is extremely rare in vegetative kingdom.Because its special physiological property, if add conventional growth hormone heteroauxin cannot promote that cauline leaf breaks up fast at low concentration in inducing culture, cause cultivating unsuccessfully at the easy bacterial infection of the overlong time of medium, if add the excessive concentration of conventional growth hormone heteroauxin, inhibit again growing of root system, cause the camel thorn root system development of tissue cultures incomplete.
Summary of the invention
Technical problem to be solved by this invention is to provide rapid induction medium of a kind of camel thorn tissue cultures and preparation method thereof.
Technical problem to be solved by this invention is achieved by the following technical programs:
A kind of rapid induction medium of camel thorn tissue cultures, described inducing culture is add following component in 1/2MS medium: special curing agent, brown sugar, 6-benzyl aminoadenine, the chloro-IAA of 4-, tetracycline, active carbon, plant extracts, Chinese flowering quince juice is formulated, concentration in inducing culture is respectively: special curing agent 15-18g/L, brown sugar 20-25g/L, tetracycline 25-45mg/L, 6-benzyl aminoadenine 2.2-2.6mg/L, the chloro-IAA 0.3-0.4mg/L of 4-, plant extracts 50-80mg/L, Chinese flowering quince juice 60-80g/L, active carbon 3-4g/L.
Further, described special curing agent: comprise sodium alginate and purple potato extract, its preparation method is, fresh purple potato is squeezed, collect juice, then juice vacuum drying is obtained the block solidified, then block is crushed to particle diameter 150-300 object powdery, be purple potato extract; Described special curing agent is that sodium alginate compares 2:3 with purple potato extract quality.
Further, described plant extracts is extracted from the aerial root of great Ye banyan, the extracting method of described plant extracts is for collecting great Ye banyan aerial root, put into agitator, and insert acetic acid-ethanol water according to mass volume ratio 1:1kg/L and carry out historrhexis, then filtrate is collected after broken liquid being crossed 300 order filter clothes, the powder again filter vacuum drying being collected afterwards acquisition is described plant extracts, wherein said acetic acid-ethanol water to be mass concentration be 45% ethanol and mass concentration be 12% acetic acid mix according to the volume ratio of 1:2.
A preparation method for the rapid induction medium of camel thorn tissue cultures, adds special curing agent, brown sugar, 6-benzyl aminoadenine, the chloro-IAA of 4-, tetracycline, active carbon in 1/2MS medium; Concentration in inducing culture is respectively special curing agent 15-18g/L, brown sugar 20-25g/L, the chloro-IAA 0.3-0.4mg/L of tetracycline 25-45mg/L, 6-benzyl aminoadenine 2.2-2.6mg/L, 4-, active carbon 3-4g/L, plant extracts 50-80mg/L, and be placed on sterilizing in high-pressure steam sterilizing pan, sterilising conditions is 113 degrees Celsius, pressure 103.42kPa, duration 25-35min, finally adds Chinese flowering quince juice wherein again.
Further, described Chinese flowering quince juice needs through 68 DEG C, the heat sterilization of duration 30min before interpolation, and after heating, its temperature is cooled to 1-3 DEG C rapidly, maintenance duration is 5-10min, can be then that 60-80g/L inserts in inducing culture according to concentration.
Further, described preparation process all operates in gnotobasis.
The present invention has following beneficial effect:
The present invention is directed to the special growth of camel thorn and physiological property, curing agent prepared by elite sodium alginate and purple potato extract.
The present invention adds tetracycline further in inducing culture, in incubation, obvious inhibitory action can be produced to the endophyte of explant further, the pollution in group training process can be reduced, but also serve promotion axillary bud sprouting, shorten the effect of time of sprouting.
The present invention adds plant extracts further in inducing culture, and the extract being extracted from great Ye banyan aerial root coordinates 4-chloro-IAA can the growth of efficient balance camel thorn axillalry bud, root and stem, and grows axillary bud sprouting and sprout and have obvious facilitation.
Apply inducing culture of the present invention and carry out cultivation camel thorn, survival rate is 98-99%, and microbiological contamination rate is 1.7-2.1%, and medium browning degree is the low 14-17% of MS medium comparatively, time shorten about 20% of sprouting.
Embodiment
Below in conjunction with specific embodiment, the present invention will be described in detail.
Embodiment:
The 1/2MS medium component that the present invention selects and consumption (consumption refers to the consumption in 1 liter of 1/2MS medium) thereof are as following table 1:
Then in 1/2MS medium, following component is added: special curing agent, brown sugar, 6-benzyl aminoadenine, the chloro-IAA of 4-, tetracycline, active carbon; Concentration in inducing culture is respectively special curing agent 15-18g/L, brown sugar 20-25g/L, the chloro-IAA 0.3-0.4mg/L of tetracycline 25-45mg/L, 6-benzyl aminoadenine 2.2-2.6mg/L, 4-, active carbon 3-4g/L, plant extracts 50-80mg/L, and be placed on sterilizing in high-pressure steam sterilizing pan, sterilising conditions is 113 degrees Celsius, pressure 103.42kPa, duration 25-35min, finally adds wherein by Chinese flowering quince juice again; Each component adds concentration and is preferably special curing agent 16g/L, brown sugar 22g/L, the chloro-IAA 0.35mg/L of tetracycline 35mg/L, 6-benzyl aminoadenine 2.4mg/L, 4-, plant extracts 65mg/L, Chinese flowering quince juice 70g/L, active carbon 4g/L.Described preparation process all operates in gnotobasis.
Described special curing agent: comprise sodium alginate and purple potato extract, its preparation method is, is squeezed by fresh purple potato, collect juice, then juice vacuum drying is obtained the block solidified, then block is crushed to particle diameter 200 object powdery, be purple potato extract; Described special curing agent is that sodium alginate compares 2:3 with purple potato extract quality.
The extracting method of described plant extracts is for collecting great Ye banyan aerial root, put into agitator, and insert acetic acid-ethanol water according to mass volume ratio 1:1kg/L and carry out historrhexis, then filtrate is collected after broken liquid being crossed 300 order filter clothes, the powder again filter vacuum drying being collected afterwards acquisition is described plant extracts, wherein said acetic acid-ethanol water to be mass concentration be 45% ethanol and mass concentration be 12% acetic acid mix according to the volume ratio of 1:2.
Described Chinese flowering quince juice needs through 68 DEG C, the heat sterilization of duration 30min before interpolation, and after heating, its temperature is cooled to 1-3 DEG C rapidly, maintenance duration is 8min, can be then that 70g/L inserts in inducing culture according to concentration.
Through applying the camel thorn recording inducing culture of the present invention and carry out cultivating, survival rate is 98-99%, and microbiological contamination rate is 1.7-2.1%, and medium browning degree is the low 14-17% of MS medium comparatively, time shorten 25-35% of sprouting.
The above embodiment only have expressed embodiments of the present invention; it describes comparatively concrete and detailed; but therefore can not be interpreted as the restriction to the scope of the claims of the present invention; in every case the technical scheme adopting the form of equivalent replacement or equivalent transformation to obtain, all should drop within protection scope of the present invention.
Claims (6)
1. the rapid induction medium of a camel thorn tissue cultures, it is characterized in that described inducing culture is add following component in 1/2MS medium: special curing agent, brown sugar, 6-benzyl aminoadenine, the chloro-IAA of 4-, tetracycline, active carbon, plant extracts, Chinese flowering quince juice is formulated, concentration in inducing culture is respectively: special curing agent 15-18g/L, brown sugar 20-25g/L, tetracycline 25-45mg/L, 6-benzyl aminoadenine 2.2-2.6mg/L, the chloro-IAA 0.3-0.4mg/L of 4-, plant extracts 50-80mg/L, Chinese flowering quince juice 60-80g/L, active carbon 3-4g/L.
2. the rapid induction medium of a kind of camel thorn tissue cultures according to claim 1, it is characterized in that described special curing agent: comprise sodium alginate and purple potato extract, its preparation method is, fresh purple potato is squeezed, collect juice, then juice vacuum drying is obtained the block solidified, then block is crushed to particle diameter 150-300 object powdery, be purple potato extract; Described special curing agent is that sodium alginate compares 2:3 with purple potato extract quality.
3. the rapid induction medium of a kind of camel thorn tissue cultures according to claim 1, it is characterized in that plant extracts is extracted from great Ye banyan aerial root, the extracting method of described plant extracts is for collecting great Ye banyan aerial root, put into agitator, and insert acetic acid-ethanol water according to mass volume ratio 1:1kg/L and carry out historrhexis, then filtrate is collected after broken liquid being crossed 300 order filter clothes, the powder again filter vacuum drying being collected afterwards acquisition is described plant extracts, wherein said acetic acid-ethanol water to be mass concentration be 45% ethanol and mass concentration be 12% acetic acid mix according to the volume ratio of 1:2.
4. the preparation method of the rapid induction medium of a kind of camel thorn tissue cultures as claimed in claim 3, is characterized in that in 1/2MS medium, add special curing agent, brown sugar, 6-benzyl aminoadenine, the chloro-IAA of 4-, tetracycline, active carbon; Concentration in inducing culture is respectively special curing agent 15-18g/L, brown sugar 20-25g/L, the chloro-IAA 0.3-0.4mg/L of tetracycline 25-45mg/L, 6-benzyl aminoadenine 2.2-2.6mg/L, 4-, active carbon 3-4g/L, plant extracts 50-80mg/L, and be placed on sterilizing in high-pressure steam sterilizing pan, sterilising conditions is 113 degrees Celsius, pressure 103.42kPa, duration 25-35min, finally adds Chinese flowering quince juice wherein again.
5. the preparation method of the rapid induction medium of a kind of camel thorn tissue cultures according to claim 4, it is characterized in that described Chinese flowering quince juice needs through 68 DEG C, the heat sterilization of duration 30min before interpolation, after heating, its temperature is cooled to 1-3 DEG C rapidly, maintenance duration is 5-10min, can be then that 60-80g/L inserts in inducing culture according to concentration.
6. the preparation method of the rapid induction medium of a kind of camel thorn tissue cultures according to claim 5, is characterized in that: described preparation process all operates in gnotobasis.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105145353A (en) * | 2015-08-08 | 2015-12-16 | 李沾云 | Rapid inducing medium device for tissue culture of Alhagi sparsifolia Shap |
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| CN101422111A (en) * | 2008-11-04 | 2009-05-06 | 中国科学院新疆生态与地理研究所 | Artificial breeding technique of perennial herb |
| CN102388733A (en) * | 2011-07-18 | 2012-03-28 | 中国科学院新疆生态与地理研究所 | Method for utilizing perennial plant Alhagi sparsifolia in arid area |
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| CN101422111A (en) * | 2008-11-04 | 2009-05-06 | 中国科学院新疆生态与地理研究所 | Artificial breeding technique of perennial herb |
| CN102388733A (en) * | 2011-07-18 | 2012-03-28 | 中国科学院新疆生态与地理研究所 | Method for utilizing perennial plant Alhagi sparsifolia in arid area |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105145353A (en) * | 2015-08-08 | 2015-12-16 | 李沾云 | Rapid inducing medium device for tissue culture of Alhagi sparsifolia Shap |
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Inventor after: Feng Qi Inventor after: Li Jianguo Inventor after: Guo Xiaoyan Inventor after: Guo Rui Inventor after: Jia Bing Inventor after: Xi Haiyang Inventor after: Yang Linshan Inventor before: Xu Weiming |
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Effective date of registration: 20160823 Address after: 730099 Gansu city of Lanzhou province Donggang West Road No. 260 Applicant after: Inst of Cold and Drought Regions Environment and Engineering, Chinese Academy of Address before: 23 No. 246523 Anhui Wanjiang Anqing Yingjiang District Road Applicant before: Xu Weiming |
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Effective date of registration: 20170511 Address after: 246000 Anhui Province, Anqing City Yingjiang District luck village 1 building two layer 16 axis Applicant after: Anqing Huizhi Technology Advisory Services Ltd. Address before: 730099 Gansu city of Lanzhou province Donggang West Road No. 260 Applicant before: Inst of Cold and Drought Regions Environment and Engineering, Chinese Academy of |
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