CN108541592A - The tissue culture mating system of kapok - Google Patents
The tissue culture mating system of kapok Download PDFInfo
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- CN108541592A CN108541592A CN201810477374.0A CN201810477374A CN108541592A CN 108541592 A CN108541592 A CN 108541592A CN 201810477374 A CN201810477374 A CN 201810477374A CN 108541592 A CN108541592 A CN 108541592A
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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
-
- 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
- A01G17/00—Cultivation of hops, vines, fruit trees, or like trees
- A01G17/005—Cultivation methods
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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
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
-
- 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
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
- A01G24/12—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material containing soil minerals
- A01G24/15—Calcined rock, e.g. perlite, vermiculite or clay aggregates
-
- 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
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
- A01G24/28—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing peat, moss or sphagnum
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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
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/30—Growth substrates; Culture media; Apparatus or methods therefor based on or containing synthetic organic compounds
-
- 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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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Cell Biology (AREA)
- Chemical & Material Sciences (AREA)
- Inorganic Chemistry (AREA)
- Soil Sciences (AREA)
- Cultivation Of Plants (AREA)
Abstract
The invention discloses a kind of tissue culture mating systems of kapok, include the following steps：（1）The acquisition and processing of explant；（2）Callus tissue culture；（3）The culture of bud；（4）Culture of rootage；（5）Seedbed management.This method realizes kapok tissue cultivating and seedling, and relative to traditional seminal propagation or propagation by grafiting, tissue cultures are more conducive to realize the engineering nursery of kapok, and the tissue culture shoot survival percent is high, and thinking has further been widened for the breeding of kapok.
Description
Technical field
The invention belongs to crops to breed field, and in particular to a kind of tissue culture mating system of kapok.
Background technology
KapokBombax malabaricum,Nickname Panzhihua, Mo Lian, red jasmine, Mo Lianhua, silk cotton, spot Mang Shu, are one
The fallen leaves megaphanerophyte that kind is grown in the torrid zone and subtropical zone, it is 10 ~ 20 meters high.Silk cotton tree is Bombacaceae deciduous tree.Extensively
It is distributed in torrid areas.Kapok nature and flavor and function：It is sweet；It is light；It is cool in nature.Heat-clearing, dampness removing, removing toxic substances.For diarrhea, dysentery, metrorrhagia, sore
Poison.Common bombax flower can be used as medicine, and the fresh common bombax flower to fall down is dried, and the common bombax flower dried has medical value, removing toxic substances heat-clearing to drive
It is cold to dry, it is cooked congee with dry kapok or Baoshang.The tree-like tall and big grandness of kapok, spring safflower are in full bloom, are excellent shade tree, shade tree
And landscape tree.Can gardens cultivation it is ornamental.Kapok growth is rapid, and material is light and soft, is used for food steamer, packing case.Bombax cotton is short
And it is soft, without song is picked up, degree of hollowness is up to 86% or more, far super artificial fiber（25%-40%）With other any natural materials, it is not easy
It is soaked in water, and resistance to pressure is strong, warmth retention property is strong, natural antibacterial, does not eat into not mould, fillable pillow, life vest.Bombax cotton is praised
It is relatively thin, the fibrous material that relatively light, degree of hollowness is higher, more warming in current natural fiber for " plant soft gold ".
Arbor of the kapok as Guangxi province preciousness, mode of reproduction are mainly that seed seedling-raising cultivates stock, are then grafted
Breeding, but due to seed oil content height, easily degenerate and lose germination, it is general to require to adopt rear current year sowing in time.
Invention content
The present invention provides a kind of tissue culture mating system of kapok, kapok tissue cultivating and seedling is realized, relative to traditional kind
Son breeding or propagation by grafiting, tissue cultivating and seedling are more conducive to realize the engineering nursery of kapok, and the tissue culture shoot survival percent is high, for
Thinking has further been widened in the breeding of kapok.
The technical proposal of the invention is realized in this way：
The tissue culture mating system of kapok, it is characterised in that：Include the following steps：
（1）The acquisition and processing of explant：When annual kapok spring bud is sprouted, acquire its tender shoots as explant, acquisition it is tender
Bud must be complete, disease-free, is 0.35% potassium permanganate with mass fraction after the tender shoots of acquisition rinses 3 ~ 4 times repeatedly with pure water
After 15 ~ 25min of solution disinfection, blotted with sterilized filter paper, it is spare.
（2）Callus tissue culture：Go out to grow the bud section of 2.5 ~ 4cm with the scissors sterilized is crosscutting, bud section is put into callus group
It knits in culture medium and cultivates, in 25 ~ 28 DEG C of temperature, humid control 80 ~ 90%, light culture 25 ~ 28 days obtains callus.
（3）The culture of bud：Under sterile working, chooses not by the callus of living contaminants, be transferred to shoot proliferation training
It supports and carries out luring bud culture on base, cultivate 40 ~ 45 days；Condition of culture：28 ~ 30 DEG C of temperature, 2800 ~ 3500lux of intensity of illumination are waited for not
Break and terminates when having axillary buds formation.
（4）Culture of rootage：The subculture sprout that sprout grows 4.5 ~ 5cm is chosen, is transferred on root media and cultivates, cultivates item
Part：28 ~ 30 DEG C of temperature, 1800 ~ 2500lux of intensity of illumination；After culture 35 ~ 38 days, main root grows 3 ~ 3.5cm, when lateral root is 5 extra
Culture terminates.
（5）Seedbed management：
A medium of seedling bed：Medium of seedling bed is by sandy loam：Humic acid：Vermiculite：Fungicide by weight ratio 50:35:15:2.5 prepare and
At matrix prepares disinfection of being exposed to the sun under the rear sun, spare.
The transplantation of seedlings of taking root that b completes culture of rootage is to seedbed, and seeding row spacing 5cm × 5cm digs cave and plants, 3 ~ 5cm of depth,
Earthing after plantation sprays root water, until drenching, is transferred at the moon and cultivates, be transferred under natural light and manage after undetermined.
C seedbed management temperature is 25 ~ 28 DEG C；Early period focuses on water and fertilizer management, waters in a manner of spraying, and keeps seedbed not flooded
It is not non-irrigated, it uses liquid nitrogen fertilizer within 20 days after transplanting, the potassium dihydrogen phosphate that mass fraction is 0.1% is sprayed after 30 days；Mid-term is noted
Grave illness Pest management, it is 25% pyraclostrobin missible oil that a mass fraction was sprayed every 10 days, dilutes 5000 times of sprayings, simultaneously
It was found that when pest damage, 1.8% abamectin emulsifiable concentrate is sprayed, dilutes 1500 ~ 1800 times of sprayings；When seedling is grown to 10 ~ 12cm high,
Tissue-cultured seedling culture finishes, and prepares transplanting to crop field.
Step（2）The formula of the callus tissue culture base is 1/2MS+N6- benzyladenine 0.15 ~ 0.18mg/L+ naphthalenes
Acetic acid 0.6 ~ 0.75mg/L+ alginic acid 22 ~ 24g/L+ sucrose 70g/L+ agar 4.0g/L, pH5.8 ~ 6.2.
Step（3）The squamous subculture based formulas be 0.38 ~ 0.4mg/L of MS+ sucrose 4.0g/L+N6- benzyladenines+
0.4 ~ 0.45mg/L of methyl α-naphthyl acetate+heteroauxin potassium 0.05 ~ 0.08mg/L+ carbendazim 0.05 ~ 0.06mg/L+ agar 5g/L, pH
6.0~6.2。
Step（4）The formula of the root media be MS+ sucrose 4.2g/L+0.08 ~ 0.12mg/L of heteroauxin potassium+
Methyl α-naphthyl acetate 0.35 ~ 0.55mg/L+IBA 0.18 ~ 0.22mg/L+ amino-oligosaccharide 0.28 ~ 0.32mg/L+ agar 5g/L,
pH6.2~6.5。
Step（5）The medium of seedling bed fungicide is a kind of in carbendazim, pyraclostrobin, thiophanate methyl or Fluoxastrobin
Or several composition.
Step（5）Nitrate nitrogen quality proportioning is not less than 35% in the liquid nitrogen fertilizer, and ammonium nitrogen quality proportioning is not less than 10%,
Amidonitrogen quality proportioning is not less than 15%.
Beneficial effects of the present invention are：Kapok is high megaphanerophyte, and tender shoots breaks up relatively slow during tissue culture, it is not easy at
Work(, the method for the present invention are optimized by a series of condition of tissue culture, and study the type of culture medium under different stages of growth, are passed through
Have reproduction speed fast after optimization, axillary bud differentiation it is fast and take root rapid and well developed root system the features such as, be the further exploitation of kapok
It lays a solid foundation.
Specific implementation mode
The tissue culture mating system of 1 kapok of embodiment, includes the following steps：
（1）The acquisition and processing of explant：When annual kapok spring bud is sprouted, acquire its tender shoots as explant, acquisition it is tender
Bud must be complete, disease-free, is 0.35% potassium permanganate with mass fraction after the tender shoots of acquisition rinses 3 ~ 4 times repeatedly with pure water
After 15 ~ 25min of solution disinfection, blotted with sterilized filter paper, it is spare.
（2）Callus tissue culture：Go out to grow the bud section of 2.5 ~ 4cm with the scissors sterilized is crosscutting, bud section is put into callus group
It knits in culture medium and cultivates, in 25 ~ 28 DEG C of temperature, humid control 80 ~ 90%, light culture 25 ~ 28 days obtains callus；Callus group
Knit culture medium formula be 1/2MS+N6- benzyladenine 0.15 ~ 0.18mg/L+, 0.6 ~ 0.75mg/L+ of methyl α-naphthyl acetate alginic acids 22 ~
24g/L+ sucrose 70g/L+ agar 4.0g/L, pH5.8 ~ 6.2.It is by following right that the present invention, which obtains callus tissue culture base as above,
Than experiment：
Optimum results show when N6- benzyladenines are in 0.6 ~ 0.75mg/L of a concentration of 0.15 ~ 0.18mg/L and methyl α-naphthyl acetate at
Seedling rate highest.
（3）The culture of bud：Under sterile working, chooses not by the callus of living contaminants, be transferred to shoot proliferation training
It supports and carries out luring bud culture on base, cultivate 40 ~ 45 days；Condition of culture：28 ~ 30 DEG C of temperature, 2800 ~ 3500lux of intensity of illumination are waited for not
Break and terminates when having axillary buds formation；Squamous subculture based formulas be 0.38 ~ 0.4mg/L of MS+ sucrose 4.0g/L+N6- benzyladenines+
0.4 ~ 0.45mg/L of methyl α-naphthyl acetate+heteroauxin potassium 0.05 ~ 0.08mg/L+ carbendazim 0.05 ~ 0.06mg/L+ agar 5g/L, pH
6.0~6.2.The optimization contrast test of subculture medium is as follows：
Pass through above-mentioned optimal screening, it may be determined that N6- benzyladenines in 0.38 ~ 0.4mg/L, 0.4 ~ 0.45mg/L of methyl α-naphthyl acetate and
Best results under heteroauxin potassium 0.05 ~ 0.08mg/L concentration, the present invention again carry out proportioning of the three under optimum combination
Optimization：
As it can be seen that the optium concentration screened above germinates more preferably for differentiation under cooperation, the combination in concentration gradient reaches
4.0 or more.
（4）Culture of rootage：The subculture sprout that sprout grows 4.5 ~ 5cm is chosen, is transferred on root media and cultivates, cultivates item
Part：28 ~ 30 DEG C of temperature, 1800 ~ 2500lux of intensity of illumination；After culture 35 ~ 38 days, main root grows 3 ~ 3.5cm, when lateral root is 5 extra
Culture terminates.The present invention is sieved by carrying out the optimization of concentration to heteroauxin potassium, methyl α-naphthyl acetate and IBA in prescription of rooting medium
Choosing, finally determines that optimum formula is：MS+ sucrose 4.2g/L+0.08 ~ 0.12mg/L of heteroauxin potassium+methyl α-naphthyl acetate 0.35 ~
0.55mg/L+IBA 0.18 ~ 0.22mg/L+ amino-oligosaccharide 0.28 ~ 0.32mg/L+ agar 5g/L, pH6.2 ~ 6.5.Formula
There is middle addition amino-oligosaccharide strengthening root effect, contrast test to find that rough is apparent after amino-oligosaccharide is added, and fibrous root is numerous
It is fast to grow speed.
（5）Seedbed management：
A medium of seedling bed：Medium of seedling bed is by sandy loam：Humic acid：Vermiculite：Fungicide by weight ratio 50:35:15:2.5 prepare and
At matrix prepares disinfection of being exposed to the sun under the rear sun, spare；In carbendazim, pyraclostrobin, thiophanate methyl or Fluoxastrobin it is a kind of or
Several compositions.In seedbed be added fungicide purpose be in order to prevent seedling in growth and development process by soil-borne disease
Influence, Experimental Comparison finds carbendazim, pyraclostrobin, thiophanate methyl and Fluoxastrobin to experimental subjects sclerotiniose, climing withered
Disease, samping off, damping-off, pythium rot and stem rot have a preferable control effect, and control effect is up to 90% or more.
The transplantation of seedlings of taking root that b completes culture of rootage is to seedbed, and seeding row spacing 5cm × 5cm digs cave and plants, 3 ~ 5cm of depth,
Earthing after plantation sprays root water, until drenching, is transferred at the moon and cultivates, be transferred under natural light and manage after undetermined.
C seedbed management temperature is 25 ~ 28 DEG C；Early period focuses on water and fertilizer management, waters in a manner of spraying, and keeps seedbed not flooded
It is not non-irrigated, use liquid nitrogen fertilizer within 20 days after transplanting, nitrate nitrogen quality proportioning is not less than 35% in liquid nitrogen fertilizer, and ammonium nitrogen quality is matched
Than being not less than 10%, amidonitrogen quality proportioning is not less than 15%.It is molten that the potassium dihydrogen phosphate that mass fraction is 0.1% is sprayed after 30 days
Liquid；Mid-term focuses on pest management, and it is 25% pyraclostrobin missible oil that a mass fraction was sprayed every 10 days, dilutes 5000 times
When spraying, while finding pest damage, 1.8% abamectin emulsifiable concentrate is sprayed, dilutes 1500 ~ 1800 times of sprayings；Wait for seedling grow to 10 ~
When 12cm high, tissue-cultured seedling culture finishes, and prepares transplanting to crop field.
Claims (6)
1. the tissue culture mating system of kapok, it is characterised in that：Include the following steps：
（1）The acquisition and processing of explant：When annual kapok spring bud is sprouted, acquire its tender shoots as explant, acquisition it is tender
Bud must be complete, disease-free, is 0.35% potassium permanganate with mass fraction after the tender shoots of acquisition rinses 3 ~ 4 times repeatedly with pure water
After 15 ~ 25min of solution disinfection, blotted with sterilized filter paper, it is spare；
（2）Callus tissue culture：Go out to grow the bud section of 2.5 ~ 4cm with the scissors sterilized is crosscutting, bud section is put into callus training
It supports and is cultivated in base, in 25 ~ 28 DEG C of temperature, humid control 80 ~ 90%, light culture 25 ~ 28 days obtains callus；
（3）The culture of bud：Under sterile working, chooses not by the callus of living contaminants, be transferred to subculture multiplication medium
On lure bud culture, cultivate 40 ~ 45 days；Condition of culture：28 ~ 30 DEG C, 2800 ~ 3500lux of intensity of illumination of temperature, waits for constantly having
Terminate when axillary buds formation；
（4）Culture of rootage：The subculture sprout that sprout grows 4.5 ~ 5cm is chosen, is transferred on root media and cultivates, condition of culture：Temperature
28 ~ 30 DEG C of degree, 1800 ~ 2500lux of intensity of illumination；After culture 35 ~ 38 days, main root grows 3 ~ 3.5cm, and lateral root is cultivated when 5 extra
Terminate；
（5）Seedbed management：
A medium of seedling bed：Medium of seedling bed is by sandy loam：Humic acid：Vermiculite：Fungicide by weight ratio 50:35:15:2.5 prepare and
At matrix prepares disinfection of being exposed to the sun under the rear sun, spare；
To seedbed, seeding row spacing 5cm × 5cm digs cave plantation, 3 ~ 5cm of depth, plantation for the transplantation of seedlings of taking root that b completes culture of rootage
After earthing, spray root water, until drenching, be transferred at the moon culture, be transferred under natural light and manage after undetermined；
C seedbed management temperature is 25 ~ 28 DEG C；Early period focuses on water and fertilizer management, waters in a manner of spraying, and keeps seedbed not flooded not
Drought uses liquid nitrogen fertilizer in 20 days after transplanting, the potassium dihydrogen phosphate that mass fraction is 0.1% is sprayed after 30 days；Mid-term is focused on
Pest management, it is 25% pyraclostrobin missible oil that a mass fraction was sprayed every 10 days, dilutes 5000 times of sprayings, sends out simultaneously
When existing pest damage, 1.8% abamectin emulsifiable concentrate is sprayed, dilutes 1500 ~ 1800 times of sprayings；When seedling is grown to 10 ~ 12cm high, group
Training seedling culture finishes, and prepares transplanting to crop field.
2. the tissue culture mating system of kapok according to claim 1, it is characterised in that：Step（2）The callus training
The formula for supporting base is 22 ~ 24g/ of 1/2MS+N6- benzyladenine 0.15 ~ 0.18mg/L+ methyl α-naphthyl acetate 0.6 ~ 0.75mg/L+ alginic acids
L+ sucrose 70g/L+ agar 4.0g/L, pH5.8 ~ 6.2.
3. the tissue culture mating system of kapok according to claim 1, it is characterised in that：Step（3）The subculture medium
Formula is 0.38 ~ 0.4mg/L of MS+ sucrose 4.0g/L+N6- benzyladenines+0.4 ~ 0.45mg/L of methyl α-naphthyl acetate+heteroauxin
Potassium 0.05 ~ 0.08mg/L+ carbendazim 0.05 ~ 0.06mg/L+ agar 5g/L, pH 6.0 ~ 6.2.
4. the tissue culture mating system of kapok according to claim 1, it is characterised in that：Step（4）The root media
Formula be MS+ sucrose 4.2g/L+0.08 ~ 0.12mg/L of heteroauxin potassium+0.35 ~ 0.55mg/L of methyl α-naphthyl acetate+IBA
0.18 ~ 0.22mg/L+ amino-oligosaccharide 0.28 ~ 0.32mg/L+ agar 5g/L, pH6.2 ~ 6.5.
5. the tissue culture mating system of kapok according to claim 1, it is characterised in that：Step（5）In the liquid nitrogen fertilizer
Nitrate nitrogen quality proportioning is not less than 35%, and ammonium nitrogen quality proportioning is not less than 10%, and amidonitrogen quality proportioning is not less than 15%.
6. the tissue culture mating system of kapok according to claim 1, it is characterised in that：Step（5）The medium of seedling bed kills
Microbial inoculum is composition one or more of in carbendazim, pyraclostrobin, thiophanate methyl or Fluoxastrobin.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109315289A (en) * | 2018-10-09 | 2019-02-12 | 云南善源生物科技发展有限公司 | Hymsleya amabilis Novel sterilizing mode |
| CN109329062A (en) * | 2018-11-27 | 2019-02-15 | 广西玉林市华睿茶业有限公司 | A kind of tissue culture technique of cottonrose hibiscus leaf |
| CN116420619A (en) * | 2023-04-06 | 2023-07-14 | 中国热带农业科学院海口实验站 | Tissue culture and rapid propagation method for safflower kapok |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109315289A (en) * | 2018-10-09 | 2019-02-12 | 云南善源生物科技发展有限公司 | Hymsleya amabilis Novel sterilizing mode |
| CN109329062A (en) * | 2018-11-27 | 2019-02-15 | 广西玉林市华睿茶业有限公司 | A kind of tissue culture technique of cottonrose hibiscus leaf |
| CN116420619A (en) * | 2023-04-06 | 2023-07-14 | 中国热带农业科学院海口实验站 | Tissue culture and rapid propagation method for safflower kapok |
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Application publication date: 20180918 |