CN101248762A - Golden leaf carex shoot-tip culture quick propagation method - Google Patents
Golden leaf carex shoot-tip culture quick propagation method Download PDFInfo
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- CN101248762A CN101248762A CNA2008100605441A CN200810060544A CN101248762A CN 101248762 A CN101248762 A CN 101248762A CN A2008100605441 A CNA2008100605441 A CN A2008100605441A CN 200810060544 A CN200810060544 A CN 200810060544A CN 101248762 A CN101248762 A CN 101248762A
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Abstract
The invention relates to a rapid culture and propagation method of Carex evergold stem tips, which belongs to the technology field of the plant propagation, including the following steps: preparing the culture medium, selecting and sterilizing the explants, induction culturing, proliferation culturing, rooting culturing and transplanting. The method has the advantages that the detergent soaking pretreatment and the two-step sterilization of the explants effectively prevent the explant inoculation from pollution and the inoculation rate reaches 85%; the optimal culturing medium of the provided Carex evergold tissue culturing is pertinent and applicable. The induction rate of the buds reaches more than 90% and the bud is more than five times the proliferation rate in each culture period. When inoculating in proliferation culturing, breaking off the multiple shoots into simple buds instead of cutting by the scalpel, the rooting portion is prevented from forming the calli, the differentiation of all-green or all-white tissue culturing is reduced, and a yield rate of more than 95% is reached. The growth rate of the buds is high with the rooting rate at 100% and the transplanting survival rate at more than 90%, which is applicable to the factory mass production.
Description
Technical field
The present invention relates to plant tissue culture fast breeding technique field, relate in particular to a kind of golden leaf carex shoot-tip culture quick propagation method.
Background technology
Gold leaf sedge (Carex evergold) belongs to for the sedge family sedge, and perennial herb is soft elegant, 20 centimetres of plant heights, leaf has striped, and the blade both sides are green limit, in be yellow, spike, 4～May of florescence, happiness light, anti-half is cloudy, intolerant to waterlogging, adaptability is strong, can be used as flower bed, flower border fringing ornamental foliage plant, also can potted plantly view and admire, help building garden landscape cheaply, its ornamental plantation is worth and causes insider's great attention.The gold leaf sedge is generally adopted division propagation at present, and reproduction speed is slow, and percentage of seedgermination is low again.
Gold leaf sedge garden-variety is introduced from Britain in the Zhejiang Academy of Agricultural Science, intends breeding seedling in a large number by plant tissue culture technique, exports American-European countries then.Promoted after can also planting experimentally at home simultaneously.Adopt method for plant tissue culture can breed various plants in a short time fast, reproduction rate height not only, and, can keep the merit of former stock because it is vegetative propagation.But the complexity of different plant utilization method for plant tissue culture breeding seedling is different, find in the group training test in the early stage of gold leaf sedge, gold leaf sedge growing point is many near soil even place into the soil, the chance that infects diseases such as various microorganisms in the soil is many, and sterilization treatment is difficulty relatively; The problems such as tissue cultivating seedling gold leaf proterties instability of golden-rimmed plant or golden heart plant when the tissue cultivating seedling enrichment culture, also occur, make it can't realize large-scale industrialized production.
Summary of the invention
The present invention will solve the defective of above-mentioned prior art, and a kind of golden leaf carex shoot-tip culture quick propagation method is provided.
The present invention solves the technical scheme that its technical problem adopts.This golden leaf carex shoot-tip culture quick propagation method, carry out as follows:
1) culture medium preparation comprises that each component and every liter of contained weight of minimal medium and each stage medium of group training is:
(1) minimal medium: select the MS medium for use, sucrose or white sugar 20～30g/L, agar 7～9g/L, pH5.6～5.8;
(2) inducing culture: MS+KT 0.5～2.0mg/L+IAA 0.05～0.2mg/L;
(3) proliferated culture medium: MS+KT 0.1～1.0mg/L+IAA 0.05～0.2mg/L;
(4) strong seedling culture base: MS+KT 0.1～0.5mg/L+IAA 0.05～0.2mg/L;
(5) root media: 1/2MS+NAA0.1～0.5mg/L+IAA 0.05～0.2mg/L;
2) explant selection and sterilization: the gold leaf sedge tiller the growth vigorous season, 9～10 points in fine morning, select disease-free good plant, get the tiller bud of high about 5～10cm, remove the blade of top elongated and the root system of bottom, be organized into and have growing point, be about a section of 1.5cm as explant, standby after sterilization treatment;
3) inducing culture: the explant after the sterilization treatment under aseptic condition, is won the stem apex of 1～2mm, is seeded on the inducing culture, under condition of culture, cultivate 30～45 days after, induce from stem apex to form the young shoot or the bud of growing thickly;
4) enrichment culture: will induce the young shoot of formation or the bud of growing thickly to break into simple bud off with the fingers and thumb, and try not to form callus to avoid the tissue cultivating seedling base portion with the scalpel cutting; Remove complete green or complete white tissue cultivating seedling then, select tissue cultivating seedling and receive on the proliferated culture medium, under condition of culture, cultivate to breed in 30～45 days and to grow thickly bud with gold leaf; According to demand to seedling quantity, every 30～45 days by carry out seedling enrichment culture again with quadrat method;
5) strong seedling culture: be inoculated on the strong seedling culture base breeding the bud of growing thickly that, under condition of culture, cultivate 20～30 days after, seedling plant height growth reaches 3～5 centimetres;
6) culture of rootage: after being cut into individual plant the strong sprout of cultivating, remove complete green or complete white tissue cultivating seedling, select tissue culture plant inoculation and in root media, carry out culture of rootage with gold leaf, under condition of culture, cultivate 20～30 days after, seedling grows tall and grows 5～10 radiculas into about 5～7cm, seedling base portion;
7) transplant: the tissue cultivating seedling of will taking root is transplanted in matrix, cultivates 1～2 month to Cheng Miao.
Golden leaf carex shoot-tip culture quick propagation method of the present invention, medium comprise that each component and every liter of contained weight of minimal medium and each stage medium of group training are:
(1) minimal medium: select the MS medium for use, agar 7g/L, pH5.8;
(2) inducing culture: MS+KT 1.0mg/L+IAA 0.1mg/L+ sucrose 30g/L;
(3) proliferated culture medium: MS+KT 0.5mg/L+IAA 0.1mg/L+ white sugar 30g/L;
(4) strong seedling culture base: MS+KT0.25mg/L+IAA 0.05mg/L+ white sugar 30g/L;
(5) root media: 1/2MS+NAA0.2mg/L+IAA 0.1mg/L+ white sugar 20g/L;
Golden leaf carex shoot-tip culture quick propagation method of the present invention, described sterilization treatment is that the explant of putting in order carries out preliminary treatment, in the aqueous solution, add weight ratio and be 5% neutral detergent and weight ratio and be 1% tween, the 5～10min that soaks and vibrate, running water flushing 20～30min; Pretreated explant is with 75% alcohol surface sterilization, 30～45s, aseptic water washing 3 times; Soak vibration sterilization 10～12min, aseptic water washing 3～5 times with the liquor natrii hypochloritis then; Soak vibration sterilization 5～7min, aseptic washing 3～5 times with 0.1% mercuric chloride solution again.
Golden leaf carex shoot-tip culture quick propagation method of the present invention, the described condition of culture of respectively organizing the training stage be, cultivation temperature is 25 ± 2 ℃, and the illumination light intensity is 2500～3000Lx, and light application time is 16h/d.
Golden leaf carex shoot-tip culture quick propagation method of the present invention, transplanting medium are by peat: perlite: vermiculite 1: 2: 2 by volume is formulated.
The invention has the beneficial effects as follows:
1), this method soaks preliminary treatment because explant has carried out washing agent, two step sterilizations have prevented explant inoculation pollution rate effectively, success ratio of inoculation reaches 85%.
2), it is with strong points that medium is optimized in the gold leaf sedge group training that proposes, applicability is good, the inductivity of bud reaches more than 90%, each cultivation cycle of the rate of increase of bud reaches more than 5 times, when enrichment culture is inoculated, do not cut with scalpel, and the bud of will growing thickly is broken into simple bud off with the fingers and thumb, avoid the tissue cultivating seedling base portion to form callus, reduce complete green or complete white tissue cultivating seedling differentiation, the pick-up rate of gold leaf tissue cultivating seedling reaches more than 95%, the seedling fast growth, rooting rate reaches 100%, transplanting survival rate is more than 90%, and year tissue cultivating seedling production capacity can reach 1,000,000 seedlings above (a year is 8 cultivation cycle), is suitable for large-scale industrialized production.
Embodiment
The present invention is described in further detail by following examples, but content of the present invention is not limited thereto.
Embodiment 1
1) culture medium preparation comprises that each component and every liter of contained weight of minimal medium and each stage medium of group training is:
(1) minimal medium: select the MS medium for use, agar 7g/L, pH5.8;
(2) inducing culture: MS+KT 1.0mg/L+IAA 0.1mng/L+ sucrose 30g/L;
(3) proliferated culture medium: MS+KT0.5mg/L+IAA 0.1mg/L+ white sugar 30g/L;
(4) strong seedling culture base: MS+KT0.25mg/L+IAA 0.05mg/L+ white sugar 30g/L;
(5) root media: 1/2MS+NAA0.2mg/L+IAA 0.1mg/L+ white sugar 20g/L;
2) explant selection and sterilization: the gold leaf sedge tiller the growth vigorous season, 9～10 points in fine morning, select disease-free good plant, get the tiller bud of high about 5～10cm, remove the blade of top elongated and the root system of bottom, be organized into and have growing point, be about a section of 1.5cm as explant, standby after sterilization treatment;
3) inducing culture: the explant after the sterilization treatment under aseptic condition, is won the stem apex of 1～2mm, is seeded on the inducing culture, under condition of culture, cultivate 30～45 days after, induce from stem apex to form the young shoot or the bud of growing thickly;
4) enrichment culture: will induce the young shoot of formation or the bud of growing thickly to break into simple bud off with the fingers and thumb, and try not to form callus to avoid the tissue cultivating seedling base portion with the scalpel cutting; Remove complete green or complete white tissue cultivating seedling then, select tissue cultivating seedling and receive on the proliferated culture medium, under condition of culture, cultivate to breed in 30～45 days and to grow thickly bud with gold leaf; According to demand to seedling quantity, every 30～45 days by carry out seedling enrichment culture again with quadrat method;
5) strong seedling culture: be inoculated on the strong seedling culture base breeding the bud of growing thickly that, under condition of culture, cultivate 20～30 days after, seedling plant height growth reaches 3～5 centimetres;
6) culture of rootage: after being cut into individual plant the strong sprout of cultivating, remove complete green or complete white tissue cultivating seedling, select tissue culture plant inoculation and in root media, carry out culture of rootage with gold leaf, under condition of culture, cultivate 20～30 days after, seedling grows tall and grows 5～10 radiculas into about 5～7cm, seedling base portion;
7) transplant: the tissue cultivating seedling of will taking root is transplanted in matrix, cultivates 1～2 month to Cheng Miao.
Described golden leaf carex shoot-tip culture quick propagation method, described sterilization treatment is that the explant of putting in order carries out preliminary treatment, in the aqueous solution, add weight ratio and be 5% neutral detergent and weight ratio and be 1% tween, the 5～10min that soaks and vibrate, running water flushing 20～30min; Pretreated explant is with 75% alcohol surface sterilization, 30～45s, aseptic water washing 3 times; Soak vibration sterilization 10～12min, aseptic water washing 3～5 times with the liquor natrii hypochloritis then; Soak vibration sterilization 5～7min, aseptic washing 3～5 times with 0.1% mercuric chloride solution again.
Described golden leaf carex shoot-tip culture quick propagation method is characterized in that, the described condition of culture of respectively organizing the training stage is, cultivation temperature is 25 ± 2 ℃, and the illumination light intensity is 2500～3000Lx, and light application time is 16h/d.
Described golden leaf carex shoot-tip culture quick propagation method is characterized in that: described transplanting medium is by peat: perlite: vermiculite 1: 2: 2 by volume is formulated.
Embodiment 2:
In this example, minimal medium: select the MS medium for use, agar 8g/L, pH5.6; Inducing culture: MS+KT 0.5mg/L+IAA 0.05mg/L+ sucrose 30g/L; Proliferated culture medium: MS+KT 0.1mg/L+IAA 0.05mg/L+ white sugar 30g/L; Strong seedling culture base: MS+KT 0.1mg/L+IAA 0.05mg/L+ white sugar 30g/L; Root media: 1/2MS+NAA0.1mg/L+IAA0.05mg/L+ white sugar 20g/L.
All the other steps, condition all are same as embodiment 1.
Embodiment 3:
In this example, minimal medium: select the MS medium for use, agar 9g/L, pH5.7; Inducing culture: MS+KT2.0mg/L+IAA0.2mg/L+ sucrose 30g/L; Proliferated culture medium: MS+KT 1.0mg/L+IAA0.2mg/L+ white sugar 30g/L; Strong seedling culture base: MS+KT 0.5mg/L+IAA 0.2mg/L+ white sugar 30g/L; Root media: 1/2MS+NAA0.5mg/L+IAA0.2mg/L+ white sugar 20g/L.
All the other steps, condition all are same as embodiment 1.
Embodiment 4:
In this example, minimal medium: select the MS medium for use, sucrose or white sugar 20～30g/L, agar 7～9g/L, pH5.6～5.8; Inducing culture: MS+KT 0.5～2.0mg/L+IAA 0.05～0.2mg/L; Proliferated culture medium: MS+KT 0.1～1.0mg/L+IAA 0.05～0.2mg/L; Strong seedling culture base: MS+KT 0.1～0.5mg/L+IAA 0.05～0.2mg/L; Root media: 1/2MS+NAA0.1～0.5mg/L+IAA 0.05～0.2mg/L.
All the other steps, condition all are same as embodiment 1.
In addition to the implementation, the present invention can also have other embodiments.All employings are equal to the technical scheme of replacement or equivalent transformation formation, all drop on the protection domain of requirement of the present invention.
Claims (5)
1, a kind of golden leaf carex shoot-tip culture quick propagation method is characterized in that: this method is carried out according to the following steps:
1) culture medium preparation comprises that each component and every liter of contained weight of minimal medium and each stage medium of group training is:
(1) minimal medium: select the MS medium for use, sucrose or white sugar 20～30g/L, agar 7～9g/L, pH5.6～5.8;
(2) inducing culture: MS+KT 0.5～2.0mg/L+IAA 0.05～0.2mg/L;
(3) proliferated culture medium: MS+KT 0.1～1.0mg/L+IAA 0.05～0.2mg/L;
(4) strong seedling culture base: MS+KT 0.1～0.5mg/L+IAA 0.05～0.2mg/L;
(5) root media: 1/2MS+NAA0.1～0.5mg/L+IAA 0.05～0.2mg/L;
2) explant selection and sterilization: select disease-free good plant, get the tiller bud of high 5～10cm, remove the blade of top elongated and the root system of bottom, be organized into and have growing point, be about a section of 1.5cm as explant, standby after sterilization treatment;
3) inducing culture: the explant after the sterilization treatment under aseptic condition, is won the stem apex of 1～2mm, is seeded on the inducing culture, under condition of culture, cultivate 30～45 days after, induce from stem apex to form the young shoot or the bud of growing thickly;
4) enrichment culture: will induce the young shoot of formation or the bud of growing thickly to break into simple bud off with the fingers and thumb, and try not to form callus to avoid the tissue cultivating seedling base portion with the scalpel cutting; Remove complete green or complete white tissue cultivating seedling then, select tissue cultivating seedling and receive on the proliferated culture medium, under condition of culture, cultivate to breed in 30～45 days and to grow thickly bud with gold leaf; According to demand to seedling quantity, every 30～45 days by carry out seedling enrichment culture again with quadrat method;
5) strong seedling culture: be inoculated on the strong seedling culture base breeding the bud of growing thickly that, under condition of culture, cultivate 20～30 days after, seedling plant height growth reaches 3～5 centimetres;
6) culture of rootage: after being cut into individual plant the strong sprout of cultivating, remove complete green or complete white tissue cultivating seedling, select tissue culture plant inoculation and in root media, carry out culture of rootage with gold leaf, under condition of culture, cultivate 20～30 days after, seedling grows tall into 5～7cm, the seedling base portion grows 5～10 radiculas;
7) transplant: the tissue cultivating seedling of will taking root is transplanted in transplanting medium, cultivates 1～2 month to Cheng Miao.
2, golden leaf carex shoot-tip culture quick propagation method according to claim 1 is characterized in that: described medium comprises that each component and every liter of contained weight of minimal medium and each stage medium of group training is:
(1) minimal medium: select the MS medium for use, agar 7g/L, pH5.8;
(2) inducing culture: MS+KT 1.0mg/L+IAA 0.1mg/L+ sucrose 30g/L;
(3) proliferated culture medium: MS+KT 0.5mg/L+IAA 0.1mg/L+ white sugar 30g/L;
(4) strong seedling culture base: MS+KT0.25mg/L+IAA 0.05mg/L+ white sugar 30g/L;
(5) root media: 1/2MS+NAA0.2mg/L+IAA 0.1mg/L+ white sugar 20g/L.
3, golden leaf carex shoot-tip culture quick propagation method according to claim 1, it is characterized in that: described sterilization treatment is that the explant of putting in order carries out preliminary treatment, in the aqueous solution, add weight ratio and be 5% neutral detergent and weight ratio and be 1% tween, immersion was also vibrated 5～10 minutes, running water flushing 20～30 minutes; Pretreated explant is with 75% alcohol surface sterilization 30～45 seconds, aseptic water washing 3 times; Soak vibration sterilization 10～12min, aseptic water washing 3～5 times with the liquor natrii hypochloritis then; Soak vibration sterilization 5～7min, aseptic washing 3～5 times with 0.1% mercuric chloride solution again.
4, golden leaf carex shoot-tip culture quick propagation method according to claim 1 is characterized in that, the described condition of culture of respectively organizing the training stage is, cultivation temperature is 25 ± 2 ℃, and the illumination light intensity is 2500～3000Lx, and light application time is 16h/d.
5, golden leaf carex shoot-tip culture quick propagation method according to claim 1 is characterized in that: described transplanting medium is by peat: perlite: vermiculite 1: 2: 2 by volume is formulated.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102293151A (en) * | 2010-06-24 | 2011-12-28 | 上海上房园艺有限公司 | Method for cultivating gold-leaf sedge |
| CN102326514A (en) * | 2011-11-01 | 2012-01-25 | 四川中测科技投资有限公司 | Tea tree explant mercury-free disinfectant and explant disinfection method |
| CN104719160A (en) * | 2015-03-17 | 2015-06-24 | 山西省农业科学院旱地农业研究中心 | Tissue-culture rapid propagation method taking seedling leaves of sedge as explants |
| CN109392716A (en) * | 2018-11-22 | 2019-03-01 | 张世燊 | A kind of and leaf regeneration system method for building up |
-
2008
- 2008-03-27 CN CN2008100605441A patent/CN101248762B/en not_active Expired - Fee Related
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102293151A (en) * | 2010-06-24 | 2011-12-28 | 上海上房园艺有限公司 | Method for cultivating gold-leaf sedge |
| CN102293151B (en) * | 2010-06-24 | 2013-03-06 | 上海上房园艺有限公司 | Method for cultivating gold-leaf sedge |
| CN102326514A (en) * | 2011-11-01 | 2012-01-25 | 四川中测科技投资有限公司 | Tea tree explant mercury-free disinfectant and explant disinfection method |
| CN104719160A (en) * | 2015-03-17 | 2015-06-24 | 山西省农业科学院旱地农业研究中心 | Tissue-culture rapid propagation method taking seedling leaves of sedge as explants |
| CN109392716A (en) * | 2018-11-22 | 2019-03-01 | 张世燊 | A kind of and leaf regeneration system method for building up |
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| CN101248762B (en) | 2010-06-02 |
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