CN105145353A - Rapid inducing medium device for tissue culture of Alhagi sparsifolia Shap - Google Patents
Rapid inducing medium device for tissue culture of Alhagi sparsifolia Shap Download PDFInfo
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- CN105145353A CN105145353A CN201510480426.6A CN201510480426A CN105145353A CN 105145353 A CN105145353 A CN 105145353A CN 201510480426 A CN201510480426 A CN 201510480426A CN 105145353 A CN105145353 A CN 105145353A
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Abstract
The invention particularly relates to a rapid inducing medium device for tissue culture of Alhagi sparsifolia Shap. The rapid inducing medium device comprises a sterile box and an inducing medium located in the sterile box, wherein the sterile box comprises an upper cubic operation box and a lower funnel-shaped culture box fixed at the bottom of the upper operation box; the inducing medium is prepared by adding a special curing agent, brown sugar, 6-benzylaminopurine, 4-chloro-IAA (indole-3-acetic acid), tetracycline, activated carbon, plant extract and papaya juice into a 1/2MS medium. According to the rapid inducing medium device, the tetracycline is further added to the inducing medium and can have an obvious inhibiting effect on endophyte of an explant, further, by means of cooperation of ficus lacor aerial root extract and 4-chloro-IAA, growth of axillary buds, roots and stems of Alhagi sparsifolia Shap can be effectively balanced, and axillary bud germination and bud development are remarkably promoted. With the application of the inducing medium for culture of Alhagi sparsifolia Shap, the rate of survival is 98%-99%, the contamination rate is 1.7%-2.1%, the browning degree of the medium is 14%-17% lower than that of an MS 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 a kind of rapid induction medium device of camel thorn tissue cultures.
Background technology
Camel thorn (formal name used at school: AlhagisparsifoliaShap.) 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
desertwith 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 a kind of rapid induction medium device of camel thorn tissue cultures.
Technical problem to be solved by this invention is achieved by the following technical programs:
A kind of rapid induction medium device of camel thorn tissue cultures, comprise sterile board and the inducing culture being positioned at sterile board, described sterile board comprises cube-shaped upper control box and is fixed on the funnelform lower incubator bottom control box, operating desk is provided with bottom upper control box, the sidepiece of operating desk is a portable plate, upper control box and lower incubator can be cut off by communication type by portable plate, upper control box is provided with three groups of gathering holes and a charging aperture, one group of gathering hole includes two handle holes, handle hole is connected with hand gum cover, hand gum cover comprises interior gum cover layer, cotton layer and outer gum cover layer, cotton layer is filled with the uniform expanded polyethylene foam of thickness, outer gum cover layer is enclosed within staff outside being close to, charging aperture is connected with the feed pipe of soft glue, feed pipe comprises interior sebific duct layer, vacuum layer and outer sebific duct layer, feed pipe end is provided with check valve, the bottom of lower incubator is platform, lower incubator sidepiece is provided with one can the wicket of free opening and closing, wicket and lower incubator are connected through the hinge, wicket surrounding is surrounded by fluid sealant, 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 are formulated, and the concentration in inducing culture is respectively: special curing agent 15-18g/L, brown sugar 20-25g/L, the chloro-IAA0.3-0.4mg/L of tetracycline 25-45mg/L, 6-benzyl aminoadenine 2.2-2.6mg/L, 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 inducing culture, 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-IAA0.3-0.4mg/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.
The present invention has following beneficial effect:
The present invention in sterile board, the growth special for 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 25-35% of sprouting.
Accompanying drawing explanation
The invention will be further described to utilize accompanying drawing, but the embodiment in accompanying drawing does not form any limitation of the invention.
Fig. 1 is the structure chart of the rapid induction medium device of camel thorn tissue cultures of the present invention.
Embodiment
Below in conjunction with specific embodiment, the present invention will be described in detail.
Embodiment:
A kind of rapid induction medium device of camel thorn tissue cultures, comprise sterile board and the inducing culture being positioned at sterile board, the funnelform lower incubator 2 that sterile board comprises cube-shaped upper control box 1 and is fixed on bottom control box 1, operating desk 9 is provided with bottom upper control box 1, the sidepiece of operating desk 9 is a portable plate 10, upper control box 1 can be cut off by communication type by portable plate 10 with lower incubator 2, upper control box 1 is provided with three groups of gathering holes and a charging aperture, one group of gathering hole includes two handle holes, handle hole is connected with hand gum cover, hand gum cover comprises interior gum cover layer, cotton layer and outer gum cover layer, cotton layer is filled with the uniform expanded polyethylene foam of thickness, outer gum cover layer is enclosed within staff outside being close to, charging aperture is connected with the feed pipe 5 of soft glue, feed pipe 5 comprises interior sebific duct layer, vacuum layer and outer sebific duct layer, feed pipe 5 end is provided with check valve, the bottom of lower incubator 2 is platform 8, lower incubator 2 sidepiece is provided with one can the wicket 6 of free opening and closing, wicket 6 is connected by hinge 7 with lower incubator 2, wicket 6 surrounding is surrounded by fluid sealant.
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-IAA0.3-0.4mg/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-IAA0.35mg/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.
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 (5)
1. the rapid induction medium device of a camel thorn tissue cultures, it is characterized in that, comprise sterile board and the inducing culture being positioned at sterile board, described sterile board comprises cube-shaped upper control box (1) and is fixed on the funnelform lower incubator (2) of control box (1) bottom, upper control box (1) bottom is provided with operating desk (9), the sidepiece of operating desk (9) is a portable plate (10), upper control box (1) and lower incubator (2) can be cut off by communication type by portable plate (10), upper control box (1) is provided with three groups of gathering holes and a charging aperture, one group of gathering hole includes two handle holes, handle hole is connected with hand gum cover, hand gum cover comprises interior gum cover layer, cotton layer and outer gum cover layer, cotton layer is filled with the uniform expanded polyethylene foam of thickness, outer gum cover layer is enclosed within staff outside being close to, charging aperture is connected with the feed pipe (5) of soft glue, feed pipe (5) comprises interior sebific duct layer, vacuum layer and outer sebific duct layer, feed pipe (5) end is provided with check valve, the bottom of lower incubator (2) is platform (8), lower incubator (2) sidepiece is provided with one can the wicket (6) of free opening and closing, wicket (5) is connected by hinge (7) with lower incubator (2), wicket (6) surrounding is surrounded by fluid sealant, 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 are formulated, and the concentration in inducing culture is respectively: special curing agent 15-18g/L, brown sugar 20-25g/L, the chloro-IAA0.3-0.4mg/L of tetracycline 25-45mg/L, 6-benzyl aminoadenine 2.2-2.6mg/L, 4-, plant extracts 50-80mg/L, Chinese flowering quince juice 60-80g/L, active carbon 3-4g/L.
2. the rapid induction medium device 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, collects juice, then juice vacuum drying is obtained the block solidified, again block is crushed to particle diameter 150-300 object powdery, is purple potato extract; Described special curing agent is that sodium alginate compares 2:3 with purple potato extract quality.
3. the rapid induction medium device 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 inducing culture as claimed in claim 3, is characterized in that, add 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-IAA0.3-0.4mg/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 inducing culture 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.
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Application publication date: 20151216 |