CN107372123B - A method of improving tulip flourish - Google Patents
A method of improving tulip flourish Download PDFInfo
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- CN107372123B CN107372123B CN201710820112.5A CN201710820112A CN107372123B CN 107372123 B CN107372123 B CN 107372123B CN 201710820112 A CN201710820112 A CN 201710820112A CN 107372123 B CN107372123 B CN 107372123B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
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Abstract
The invention discloses a kind of methods for improving tulip flourish, it is characterized in that, including following aspect: preparing different culture medium stage by stage, be added in the culture medium that germinates and promote plant cell division ingredient, be added in growth medium and promote tulip seedling root, leaf, Stem nematode ingredient；It is designed by taper culture bottle and incubator, tulip cultured tissue is fixed on solid-state germination culture medium, fresh medium is supplemented by taper culture bottle bottom mesoporous, culture solution in incubator supplements culture solution in incubator by inlet opening, fluid hole, replaces；Seedling after tissue cultures transplants preceding root package solid-state transplanting culture medium, discarded growth medium is re-used, while providing a variety of nutrition, antimicrobial component for plant soil transferring.
Description
Technical field
The invention belongs to technical field of flower cultivation, and in particular to a method of improve tulip flourish.
Background technique
Plantation is the state of the states such as Holland, New Zealand, Iran, Turkey, Turkmenistan extensively in tulip world wide
Flower, has become fashion and international representative, is national foreign affairs activity, large-scale celebration, China and foreign countries' flower show, high-grade palace place weight
Want component part.The features such as tulip is with its colorful pattern, florescence length, elegant, is favored by people；Due to Radix Curcumae
Fragrant condition of compatibility is relatively wide, and China south and north, which has relatively, has large-scale plantation and tulip to be the theme the construction of ornamental garden, beautification
Living environment, increases vacation tourism landscape.It is cold-resistant and thermo-labile but since tulip seed balls are expensive, to temperature
Change sensitivity is high, is restricted tulip plantation development；It is bred by tissue culture technique, it is difficult since culture solution is fixed
To supplement fresh medium, different tissues growth phase uses identical component culture medium, causes tissue cultures survival rate low；And root
Different culture solutions are used according to the tissue growth stage, increase toxigenic capacity, repeatedly transplanting can also damage tissue cultures.
Summary of the invention
To solve the above problems, the present invention provides a kind of methods for improving tulip flourish: tulip tissue connects
When kind, using solid-state germination culture medium, promote histocyte division and germinating growth；It is set by taper culture bottle and incubator
Meter can provide different role culture solution, and pass through incubator inlet opening, fluid hole according to needed for tissue different stages of growth,
Replacement original fluid in time supplements fresh medium, is more conducive to promote tissue growth；Before soil cultivation, transplanted using solid-state
Culture medium wraps up tulip seedling root, can both re-use former grown cultures waste liquid, save the cost, and can be plant soil
Abundant nutrition and antimicrobial component are provided after transplanting.
The present invention is achieved by the following technical solutions:
A method of tulip flourish being improved, following steps:
(1) bulb pre-processes: the bulb after harvest being placed in insulating box, temperature is 30 DEG C -35 DEG C, the holding time 7
It；Then, the calorstat temperature for placing bulb is reduced to 18 DEG C -22 DEG C, humidity 72%-80%, placed 45 days, the preliminary embryo of bulb
Bud germination；According to 100-120 parts of quantity count water, 0.1-0.2 parts of 25% metalaxyl wettable powder, fungicide A is prepared, it will be first
The tulip seed balls for walking germination, are placed in fungicide A, impregnate 40min；Then, the bulb after sterilization is placed in sterile board
In；
(2) it sterilizes: transfer room is closed, using in 95% ethyl alcohol, wiping transfer room workbench, instrument surface and may
Contact portion, and open ultraviolet radiator and 4-5h is irradiated to transfer room；Tweezers, test tube, vaccinating lancet, conical flask culture bottle etc. are inoculated with
Used tool, vessel, are placed in high-pressure sterilizing pot, and 121 DEG C of temperature, high pressure sterilization 25-30min；Using N6 culture medium as substrate,
Portugal's sweet dew glue, zeatin, gibberellin is added, is configured to germination culture medium B；Using N6 culture medium as substrate, 2,4- dichloro-benzenes is added
Fluoroacetic acid, gibberellin, 6- benzylaminopurine, are configured to growth medium C；To germinate culture medium B, growth medium C, place
It sterilizes in high-pressure sterilizing pot, 120 DEG C of temperature setting, time 20-25min；
(3) tissue is inoculated with: entering operating room, personnel wear the sterile clothes, mask, cap etc. by sterilizing；Germination is trained
Feeding base is placed in water-bath, and temperature is 70 DEG C -72 DEG C, and germination solid medium melt into glue is spare；Aseptic operating platform is opened,
By vaccinating lancet in alcolhol burner calcination, after cooling, tulip is splitted using vaccinating lancet rapidly and is tentatively germinateed bulb, plumule is found, it will
Plumule, which is used, to be cut into 5-7 and saves in tissue cultures；By germination culture medium gluey in water-bath, it is added in taper culture bottle, is formed
Substrate 0.6-1cm thickness culture medium rapidly moves to the plumule of interception on germination culture medium, and by conical flask bottom mesoporous and trains
Base is supported, penetrates through the hole 2-3 using glass bar；Then, it by taper culture bottle, is placed in incubator on culturing rack, covers outer cover,
Carry out tissue cultures；
Step (3) is described, taper culture bottle design size are as follows:
Shape is that upper bottom is circular conically shaped body, and tapered open end diameter is 4.5-5.0cm, and lower mouth end diameter is
2.5-3cm is highly 5-6cm, bottom be in poroid separation layer, median pore diameter 0.1-0.2cm；
Step (3) is described, incubator composition and design size are as follows:
Incubator long 55-60cm, wide 25-30cm, are highly 10-12cm；Inside there is conical flask rack, there is cloche in outside
Closing, cloche is having a size of 55.1-60.1cm, wide 25.1-30.1cm, high 2-3cm；There is 1 inlet opening at incubator both ends respectively
And fluid hole, the diameter in hole are 0.2-0.3cm；
(4) incubator tissue cultures: in incubator use daylight light irradiation, intensity of illumination 520-570lux；To strongly fragrant
After Jin Xiang organizes initial development of the market to grow up, incubator fluid hole is closed, grown cultures base fluid is entered in incubator by inlet opening,
Make grown cultures base fluid height 1.4-1.6cm in incubator, and fluorescent lamp intensity of illumination is promoted to 750-800lux；It uses
15%-20% is discharged in original fluid in incubator, and the fresh life of 15%-20% is supplemented by inlet opening simultaneously within fluid hole every 4-5 days
Long liquid medium will displace grown cultures base fluid and collect storage, is placed in insulating box, saves under 2 DEG C of -3 DEG C of low temperature；
(5) tissue sprigging: according to each 1/2 proportional quantity of ingredient of N6 original culture medium, to the grown cultures base fluid displaced
Middle addition, and Portugal's sweet dew glue, penicillin, nimbin, 2,4- dichlorphenoxyacetic acid is added, it is configured to solid-state transplanting culture medium；It will
Solid-state transplanting culture medium is wrapped in tulip seedling root, is wrapped up using cotton fine-structure mesh, carries out soil transferring cultivation, planting depth
For 4-5cm, plant spacing is 12-15cm.
Further, step (2) the germination culture medium B, is respectively configured to a point quantity count are as follows:
104 parts of N6 culture medium bottom, 0.52 part of Portugal's sweet dew glue, 0.12 part of zeatin, 0.14 part of gibberellin.
Further, step (2) the germination culture medium C, is respectively configured to a point quantity count are as follows:
N6 culture medium is 102 parts of substrate, and 2,4- 0.06 part of dichlorphenoxyacetic acids, 0.12 part of gibberellin, 6- benzylamino is fast
0.12 part of purine.
Further, step (5) the transplanting solid medium, is respectively configured to a point quantity count are as follows:
105 parts of the grown cultures base fluid displaced, each 50% proportional quantity of ingredient of N6 original culture medium, 0.64 part of Portugal's sweet dew glue are green
0.52 part of mycin, 0.35 part of nimbin, 2,4- 0.12 part of dichlorphenoxyacetic acids, 0.12 part of gibberellin.
The present invention mutually has technology to have the advantage that through taper culture bottle and incubator design, can be according to tissue not
With needed for growth phase, providing different role culture solution, and by incubator inlet opening, fluid hole, original fluid is replaced in time,
Fresh medium is supplemented, is more conducive to promote tissue growth；Before soil cultivation, solid-state transplanting culture medium package tulip children is used
Seedling root can both re-use former grown cultures waste liquid, save the cost, and can be to provide abundant nutrition after plant soil transferring
And antimicrobial component.
Specific embodiment
Embodiment 1:
(1) bulb germinates: the bulb after harvest being placed in insulating box, temperature is 32 DEG C, the holding time 7 days；Then,
The calorstat temperature for placing bulb is reduced to 19 DEG C, humidity 75% is placed 45 days, the preliminary plumule germination of bulb；
(2) bulb sterilization processing: using metalaxyl wettable powder preparation fungicide A, the tulip seed balls that will tentatively germinate,
It is placed in fungicide A, impregnates 40min；Then, the bulb after sterilization is placed in sterile board；Step (2) is described, sterilization
Agent A prepares ingredient, quality meter part: water 102kg, 25% metalaxyl wettable powder 0.1kg；
(3) transfer room is sterilized: transfer room is closed, using in 95% ethyl alcohol, wipe transfer room workbench, instrument surface
And possible contact portion, and open ultraviolet radiator and 4h is irradiated to transfer room；By tweezers, test tube, vaccinating lancet, conical flask culture bottle etc.
It is inoculated with used tool, vessel, is placed in high-pressure sterilizing pot, 121 DEG C of temperature, high pressure sterilization 27min；
(4) culture medium is prepared and sterilizes: using N6 culture medium as substrate, Portugal's sweet dew glue, zeatin, gibberellin is added, prepares
At germination culture medium B；Using N6 culture medium as substrate, 2,4- dichlorphenoxyacetic acid, gibberellin, 6- benzylaminopurine is added, matches
Growth medium C is made；To germinate culture medium B, growth medium C, be placed in high-pressure sterilizing pot and sterilize, temperature setting 120
DEG C, time 22min；
Step (4) is described, and germination culture medium B prepares ingredient, quality meter part are as follows:
N6 culture medium bottom 103kg, Portugal sweet dew glue 0.52kg, zeatin 0.11kg, gibberellin 0.13kg；
Step (4) is described, and growth medium C prepares ingredient, quality meter part are as follows:
N6 culture medium is substrate 103kg, 2,4- dichlorphenoxyacetic acid 0.062kg, and gibberellin 0.11kg, 6- benzylamino is fast
Purine 0.12kg；
(5) tissue is inoculated with: entering operating room, personnel wear the sterile clothes, mask, cap etc. by sterilizing；Germination is trained
Feeding base is placed in water-bath, and temperature is 70 DEG C, and germination solid medium melt into glue is spare；Aseptic operating platform is opened, will be connect
Kind of knife is in alcolhol burner calcination, after cooling, split tulip using vaccinating lancet rapidly and tentatively germinates bulb, plumule is found, by plumule
It uses and is cut into 7 and saves in tissue cultures；By germination culture medium gluey in water-bath, it is added in taper culture bottle, forms substrate
0.8cm thickness culture medium rapidly moves to the plumule of interception on germination culture medium, and by conical flask bottom mesoporous and culture medium
Layer penetrates through 2 holes using glass bar；Then, it by taper culture bottle, is placed in incubator on culturing rack, covers outer cover, carry out group
Knit culture；
Step (5) is described, taper culture bottle design size are as follows:
Shape is that upper bottom is circular conically shaped body, and tapered open end diameter is 4.6cm, and lower mouth end diameter is 2.6cm,
Height be 5.5cm, bottom be in poroid separation layer, median pore diameter 0.1cm；
Step (5) is described, incubator composition and design size are as follows:
Incubator long 56cm, wide 27cm, are highly 10.5cm；Inside there is conical flask rack, there are cloche closing, glass in outside
Glass cover is having a size of 56.1cm, wide 27.1cm, high 2cm；There are 1 inlet opening and fluid hole in incubator both ends respectively, and the diameter in hole is
0.2cm；
(6) incubator tissue cultures: in incubator use daylight light irradiation, intensity of illumination 540lux；To tulip
After organizing initial development of the market growth, incubator fluid hole is closed, grown cultures base fluid is entered in incubator by inlet opening, makes to train
Supporting grown cultures base fluid height in case is 1.4cm, and fluorescent lamp intensity of illumination is promoted to 780lux；Every 5 days using fluid hole
Liquid medium is newly grown by inlet opening supplement 18% by original fluid discharge 18% in incubator, and simultaneously, growth will be displaced
Liquid medium collects storage, is placed in insulating box and saves under 2.5 DEG C of low temperature；
(7) tissue sprigging: according to each 1/2 proportional quantity of ingredient of N6 original culture medium, to the grown cultures base fluid displaced
Middle addition, and Portugal's sweet dew glue, penicillin, nimbin, 2,4- dichlorphenoxyacetic acid is added, it is configured to solid-state transplanting culture medium；It will
Solid-state transplanting culture medium is wrapped in tulip seedling root, is wrapped up using cotton fine-structure mesh, carries out soil transferring cultivation, planting depth
For 4.2cm, plant spacing is 13cm；Step (7) is described, transplants solid medium group ingredient and quality meter part are as follows:
Each 50% proportional quantity of ingredient of grown cultures base fluid 104kg, N6 original culture medium displaced, Portugal sweet dew glue 0.68kg are green
Mycin 0.57kg, nimbin 0.36kg, 2,4- dichlorphenoxyacetic acid 0.14kg, gibberellin 0.15kg.
Embodiment 2:
(1) bulb germinates: the bulb after harvest being placed in insulating box, temperature is 34 DEG C, the holding time 7 days；Then,
The calorstat temperature for placing bulb is reduced to 20 DEG C, humidity 78% is placed 45 days, the preliminary plumule germination of bulb；
(2) bulb sterilization processing: using metalaxyl wettable powder preparation fungicide A, the tulip seed balls that will tentatively germinate,
It is placed in fungicide A, impregnates 40min；Then, the bulb after sterilization is placed in sterile board；Step (2) is described, sterilization
Agent A prepares ingredient, quality meter part: water 108kg, 25% metalaxyl wettable powder 0.18kg；
(3) transfer room is sterilized: transfer room is closed, using in 95% ethyl alcohol, wipe transfer room workbench, instrument surface
And possible contact portion, and open ultraviolet radiator and 5h is irradiated to transfer room；By tweezers, test tube, vaccinating lancet, conical flask culture bottle etc.
It is inoculated with used tool, vessel, is placed in high-pressure sterilizing pot, 121 DEG C of temperature, high pressure sterilization 28min；
(4) culture medium is prepared and sterilizes: using N6 culture medium as substrate, Portugal's sweet dew glue, zeatin, gibberellin is added, prepares
At germination culture medium B；Using N6 culture medium as substrate, 2,4- dichlorphenoxyacetic acid, gibberellin, 6- benzylaminopurine is added, matches
Growth medium C is made；To germinate culture medium B, growth medium C, be placed in high-pressure sterilizing pot and sterilize, temperature setting 120
DEG C, time 23min；
Step (4) is described, and germination culture medium B prepares ingredient, quality meter part are as follows:
N6 culture medium bottom 108kg, Portugal sweet dew glue 0.64kg, zeatin 0.14kg, gibberellin 0.18kg；
Step (4) is described, and growth medium C prepares ingredient, quality meter part are as follows:
N6 culture medium is substrate 107kg, 2,4- dichlorphenoxyacetic acid 0.07kg, and gibberellin 0.18kg, 6- benzylamino is fast
Purine 0.16kg；
(5) tissue is inoculated with: entering operating room, personnel wear the sterile clothes, mask, cap etc. by sterilizing；Germination is trained
Feeding base is placed in water-bath, and temperature is 72 DEG C, and germination solid medium melt into glue is spare；Aseptic operating platform is opened, will be connect
Kind of knife is in alcolhol burner calcination, after cooling, split tulip using vaccinating lancet rapidly and tentatively germinates bulb, plumule is found, by plumule
It uses and is cut into 7 and saves in tissue cultures；By germination culture medium gluey in water-bath, it is added in taper culture bottle, forms substrate
0.9cm thickness culture medium rapidly moves to the plumule of interception on germination culture medium, and by conical flask bottom mesoporous and culture medium
Layer penetrates through 2 holes using glass bar；Then, it by taper culture bottle, is placed in incubator on culturing rack, covers outer cover, carry out group
Knit culture；
Step (5) is described, taper culture bottle design size are as follows:
Shape is that upper bottom is circular conically shaped body, and tapered open end diameter is 4.8cm, and lower mouth end diameter is 3cm, high
Degree be 5.5cm, bottom be in poroid separation layer, median pore diameter 0.1cm；
Step (5) is described, incubator composition and design size are as follows: incubator long 58cm, wide 27cm, are highly 11cm；It is interior
There is conical flask rack, there is cloche closing in outside, and cloche is having a size of 58.1cm, wide 27.1cm, high 2cm；Incubator both ends
There are 1 inlet opening and fluid hole respectively, the diameter in hole is 0.2cm；
(6) incubator tissue cultures: in incubator use daylight light irradiation, intensity of illumination 560lux；To tulip
After organizing initial development of the market growth, incubator fluid hole is closed, grown cultures base fluid is entered in incubator by inlet opening, makes to train
Supporting grown cultures base fluid height in case is 1.5cm, and fluorescent lamp intensity of illumination is promoted to 790lux；Every 5 days using fluid hole
Liquid medium is newly grown by inlet opening supplement 20% by original fluid discharge 20% in incubator, and simultaneously, growth will be displaced
Liquid medium collects storage, is placed in insulating box, saves under 3 DEG C of low temperature；
(7) tissue sprigging: according to each 1/2 proportional quantity of ingredient of N6 original culture medium, to the grown cultures base fluid displaced
Middle addition, and Portugal's sweet dew glue, penicillin, nimbin, 2,4- dichlorphenoxyacetic acid is added, it is configured to solid-state transplanting culture medium；It will
Solid-state transplanting culture medium is wrapped in tulip seedling root, is wrapped up using cotton fine-structure mesh, carries out soil transferring cultivation, planting depth
For 5cm, plant spacing is 14cm；Step (7) is described, transplants solid state rheology based component and quality meter part are as follows:
Each 50% proportional quantity of ingredient of grown cultures base fluid 109kg, N6 original culture medium displaced, Portugal sweet dew glue 0.72kg are green
Mycin 0.65kg, nimbin 0.42kg, 2,4- dichlorphenoxyacetic acid 0.18kg, gibberellin 0.18kg.
Comparison 1:
This comparison 1 compared with embodiment 1, do not carry out step (4) germination culture medium prepare and use, other embodiments with
Embodiment 1 is identical.
Comparison 2:
This comparison 2 compared with embodiment 2, do not carry out step (4) growth medium prepare and use, other embodiments with
Embodiment 2 is identical.
Comparison 3:
This comparison 3 compared with embodiment 1, do not carry out step (7) transplanting culture medium prepare and use, other embodiments with
Embodiment 1 is identical.
Comparison 4:
This comparison 4 does not carry out taper culture bottle and incubator in step (5) and uses compared with embodiment 2, other embodiment party
Case is same as Example 2.
Control group:
Using N6 culture medium, germination culture medium, growth medium, transplanting culture medium and incubator of the present invention and cone is not used
Shape culture bottle.
Evaluation method: to embodiment 1, embodiment 2, comparison 1, comparison 2, comparison 3, comparison 4 and control group, in tissue cultures
Stage, soil transferring stage count its survival rate of plant, and at tulip growth of seedling 1 month, measure its growing height；Respectively
Embodiment sample size is 300 plants.
Experimental project:
Experimental result:
Compare with control group: each embodiment and comparison scheme in tissue cultures survival rate, are migrated to compared with the control group
Control group data are above in motility rate and growing height data.
Configuration proportion comparison: embodiment 1 and embodiment 2, germination culture medium, growth medium and transplanting culture medium prepare ratio
Example is different, but in parameter area, all data indifference.
The culture medium that germinates compares: compared with embodiment 1, germination culture medium, the relatively number of embodiment 1 is not used in comparison 1 for comparison 1
According to, tissue cultures survival rate low 4.3%, the low 2.2cm of growing height.
Growth medium comparison: comparison 2 compares 2 unused growth mediums, the opposite number of embodiment 2 compared with embodiment 2
According to, tissue cultures survival rate low 4%, the low 3.2cm of growing height.
Transplant culture medium comparison: compared with embodiment 1, transplanting culture medium, the opposite number of embodiment 1 is not used in comparison 3 for comparison 3
According to, transplanting success relatively low 4.4%, the low 3.6cm of growing height.
Incubator and the comparison of taper culture bottle: incubator and taper training is not used in comparison 4 compared with embodiment 2, in comparison 4
Bottle is supported, compared with 2 data of embodiment, tissue cultures survival rate low 6.4%, the low 2.8cm of growing height.
Synthesis result: using germination culture medium, growth medium and incubator, different phase uses heterogeneity culture
Fresh culture is replenished in time in base, and tulip tissue cultures survival rate and growth speed can be improved；In soil transferring, use
Transplanting culture medium can increase the supply of tulip plant nutrition, improve transplanting success and the speed of growth.
Claims (1)
1. a kind of method for improving tulip flourish, which comprises the following steps:
(1) bulb pre-processes: the bulb after harvest being placed in insulating box, temperature is 30 DEG C -35 DEG C, the holding time 7 days；So
Afterwards, the calorstat temperature for placing bulb is reduced to 18 DEG C -22 DEG C, humidity 72%-80%, placed 45 days, the preliminary plumule hair of bulb
Bud；According to 100-120 parts of quantity count water, 0.1-0.2 parts of 25% metalaxyl wettable powder, fungicide A is prepared, will tentatively be sent out
The tulip seed balls of bud are placed in fungicide A, impregnate 40min；Then, the bulb after sterilization is placed in sterile board；
(2) it sterilizes: transfer room is closed, use 95% ethanol transfer room workbench, instrument surface and possible contact portion
Point, and open ultraviolet radiator and 4-5h is irradiated to transfer room；Tweezers, test tube, vaccinating lancet, conical flask culture bottle are inoculated with institute's recruitment
Tool, vessel, are placed in high-pressure sterilizing pot, 121 DEG C of temperature, high pressure sterilization 25-30min；Using N6 culture medium as substrate, Portugal is added
Sweet dew glue, zeatin, gibberellin are configured to germination culture medium B；Using N6 culture medium as substrate, addition 2,4- dichlorphenoxyacetic acid,
Gibberellin, 6- benzylaminopurine, are configured to growth medium C；To germinate culture medium B, growth medium C, be placed in high pressure
It sterilizes in autoclave, 120 DEG C of temperature setting, time 20-25min；
(3) tissue is inoculated with: entering operating room, personnel wear sterile clothes, mask, cap by sterilizing；Germination culture medium is set
In water-bath, temperature is 70 DEG C -72 DEG C, and germination solid medium melt into glue is spare；Aseptic operating platform is opened, will be inoculated with
Knife is in alcolhol burner calcination, after cooling, split tulip using vaccinating lancet rapidly and tentatively germinates bulb, find plumule, plumule is used
Knife is cut into 5-7 and saves in tissue cultures；By germination culture medium gluey in water-bath, it is added in taper culture bottle, forms substrate
0.6-1cm thickness culture medium rapidly moves to the plumule of interception on germination culture medium, and by conical flask bottom mesoporous and culture medium
Layer penetrates through the hole 2-3 using glass bar；Then, it by taper culture bottle, is placed in incubator on culturing rack, covers outer cover, carry out
Tissue cultures；
Step (3) the taper culture bottle design size are as follows:
Shape is that upper bottom is circular conically shaped body, and tapered open end diameter is 4.5-5.0cm, and lower mouth end diameter is 2.5-
3cm is highly 5-6cm, bottom be in poroid separation layer, median pore diameter 0.1-0.2cm；
Step (3) the incubator composition and design size are as follows:
Incubator long 55-60cm, wide 25-30cm, are highly 10-12cm；Inside there is conical flask rack, there is cloche envelope in outside
It closes, cloche is having a size of 55.1-60.1cm, wide 25.1-30.1cm, high 2-3cm；Incubator both ends have respectively 1 inlet opening and
Fluid hole, the diameter in hole are 0.2-0.3cm；
(4) incubator tissue cultures: in incubator use daylight light irradiation, intensity of illumination 520-570lux；To tulip
After organizing initial development of the market growth, incubator fluid hole is closed, grown cultures base fluid is entered in incubator by inlet opening, makes to train
Supporting grown cultures base fluid height in case is 1.4-1.6cm, and fluorescent lamp intensity of illumination is promoted to 750-800lux；Use liquid out
15%-20% is discharged in every 4-5 days in original fluid in incubator by hole, and supplements the training of 15%-20% fresh Growth by inlet opening simultaneously
Base fluid is supported, grown cultures base fluid will be displaced and collect storage, be placed in insulating box, saved under 2 DEG C of -3 DEG C of low temperature；
(5) tissue sprigging: according to each 1/2 proportional quantity of ingredient of N6 original culture medium, add into the grown cultures base fluid displaced
Enter, and Portugal's sweet dew glue, penicillin, nimbin, 2,4- dichlorphenoxyacetic acid is added, is configured to solid-state transplanting culture medium；By solid-state
Transplanting culture medium is wrapped in tulip seedling root, is wrapped up using cotton fine-structure mesh, carries out soil transferring cultivation, planting depth 4-
5cm, plant spacing are 12-15cm；
Step (2) the germination culture medium B, by following quantity count at being grouped as:
Bottom 100-110 parts of N6 culture medium, 0.5-0.7 parts of Portugal's sweet dew glue, 0.1-0.15 parts of zeatin, 0.1-0.2 parts of gibberellin；
Step (2) the growth medium C, by following quantity count at being grouped as:
N6 culture medium is substrate 100-110 parts, 2,4- 0.06-0.1 parts of dichlorphenoxyacetic acids, 0.1-0.2 parts of gibberellin, 6- benzyl
0.1-0.2 parts of adenine phosphate；
Transplanting solid medium described in the step (5), have following quantity count at being grouped as:
100-110 parts of the grown cultures base fluid displaced, each 50% proportional quantity of ingredient of N6 original culture medium, Portugal sweet dew glue 0.6-0.8
Part, 0.5-0.7 parts of penicillin, 0.3-0.5 parts of nimbin, 2,4- 0.1-0.2 parts of dichlorphenoxyacetic acids, 0.1-0.2 parts of gibberellin.
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Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
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