CN104488703B - A kind of tissue culture and rapid propagation method of gold leaf Acer negundo. L - Google Patents
A kind of tissue culture and rapid propagation method of gold leaf Acer negundo. L Download PDFInfo
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- CN104488703B CN104488703B CN201410684843.8A CN201410684843A CN104488703B CN 104488703 B CN104488703 B CN 104488703B CN 201410684843 A CN201410684843 A CN 201410684843A CN 104488703 B CN104488703 B CN 104488703B
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Abstract
The invention discloses a kind of tissue culture and rapid propagation method of gold leaf Acer negundo. L, belong to field of plant tissue culture technique.It comprises explant sterilization, Primary culture, Initial culture, squamous subculture, the step of transplanting.A kind of medium is only used: 1/2DKW+0.1mg/LNAA+0.05mg/LIAA+ sucrose 30g/L+ agar 5g/L in the present invention, same medium can carry out breeding and culture of rootage simultaneously, cultivate simple flow, operate simple and easy, there is reproduction coefficient high, breeding cycle is short, does not need hardening, the feature that survival rate is high.
Description
Technical field
The invention belongs to field of plant tissue culture technique, relate to a kind of tissue culture method, be specifically related to a kind of tissue culture and rapid propagation method of gold leaf Acer negundo. L.
Background technology
Gold leaf Acer negundo. L (Acernegundo ' Aurea ') is Aceraceae Aceraceae.Gold leaf Acer negundo. L is the high megaphanerophyte of fallen leaves, is the cultivar of Acer negundo. L, introduces from Europe.Gold leaf Acer negundo. L winglike compound leaf is very large, and leaf look soft, and spring is in golden yellow; The not burnt limit of Leaves in Summer; Cold tolerance is strong, ability ﹣ 40 degree to ﹣ 45 degree of low temperature; There is wider Bioclimatic analysis, all can plant in areas of Shenyang, China northeast, Jiangsu and Zhejiang Provinces, South China, the soil of happiness good air permeability, isolated planting mass-planting; Have vigorous growth potential and rudiment power, year increment can reach two or three meter, and damage by disease and insect is few; Also can prune in greening process, hedgerow is beautified the environment.
But gold leaf Acer negundo. L natural propagation rate is low, adopts cottage propagation more.The propagation by grafiting technology of gold leaf Acer negundo. L is provided in periodical " rural science and technology " 12 phases the 55th page in 2010 " gold leaf Acer negundo. L propagation by grafiting technology "; this technology is subject to restriction in season; and final-period management flow process is loaded down with trivial details, cuttage seeding survival rate is low, is difficult to large-scale production.Therefore can adopt group culturation rapid propagating technology, obtain a large amount of seedling in a short time.
At present, tissue culture technology about gold leaf Acer negundo. L has had a small amount of report, periodical " Henan Agricultural Sciences " 2008 07 interim " gold leaf Acer negundo. L Study on tissue culture " mainly have studied the impact that variable concentrations plant growth regulating substance grows gold leaf Acer negundo. L plantlet in vitro, determine medium WPM+IBA0.08mg/L, growth coefficient is 3.11, rooting rate reaches 100%, survival rate 90%.But detailed all not for the research of gold leaf Acer negundo. L tissue culture technology, and existence group trains the problems such as flow process is more complicated.
Summary of the invention
The present invention utilizes tissue culture technique, obtain gold leaf Acer negundo. L, reproduction coefficient is 3 ~ 6, group training simple flow, time saving and energy saving, solve the problem of prior art group training flow process complexity, provide a kind of tissue culture and rapid propagation method of gold leaf Acer negundo. L, for gold leaf Acer negundo. L factorial seedling growth provides technical support.
First the present invention utilizes plant tissue culture technique to obtain the aseptic plantlet in vitro of gold leaf Acer negundo. L, then by cutting stem section, directly induction stem section is taken root, and axillary bud sprouting propagation, breeds, strong sprout, take root simultaneously.
Object of the present invention is achieved through the following technical solutions: a kind of tissue culture and rapid propagation method of gold leaf Acer negundo. L, and it comprises the following steps:
(1) explant sterilization: gather without the gold leaf Acer negundo. L tender stem segments of damage by disease and insect, remove blade, stem section is cut into 2 ~ 3cm stem with bud, on superclean bench, with alcohol sterilizing 30 ~ 60s, mercuric chloride sterilizing 5 ~ 8min, with aseptic water washing 4 ~ 6 times;
(2) Primary culture: be inoculated into by the stem with bud after sterilizing in Primary culture base, cultivates 2 ~ 3 weeks in culturing room, and axillalry bud extends 2 ~ 4cm;
(3) Initial culture: axillalry bud is cut, be inoculated in Initial culture base, cultivate 2 ~ 3 weeks in culturing room, obtain the healthy and strong aseptic plantlet in vitro of gold leaf Acer negundo. L of taking root, cultivate after 3 ~ 4 weeks, plant strain growth 6 ~ 7cm, number of taking root is 2 ~ 12, long 3 ~ the 5cm of root, regrow out at axil place axillalry bud, growth coefficient 3 ~ 6;
(4) squamous subculture: with the aseptic plantlet in vitro of gold leaf Acer negundo. L of stalwartness for female parent, its stem section is cut into 3 ~ 6 length of tape leaf stem sections, is inoculated in subculture medium, cultivates 2 ~ 3 weeks, obtains the whole plant of gold leaf Acer negundo. L aseptic seedling;
(5) transplant: wash the medium on taking root gold leaf Acer negundo. L aseptic seedling strong off, be directly transplanted on green house seedbed, controlling canopy temperature is 20 ~ 30 DEG C.
Step (2), step (3) and the medium described in step (4) are 1/2DKW+0.1mg/LNAA+0.05mg/LIAA+ sucrose 30g/L+ agar 5g/L, pH5.8 ~ 6.3.
Step (2), step (3) and the cultivation indoor conditions described in step (4) are: intensity of illumination 2600LX ~ 3000LX, light application time 14 ~ 18h, temperature 22 ~ 26 DEG C.
In described transplanting process, matrix used is perlite and humus mixing, and both weight ratios are 1:1.5 ~ 2.5; With commercially available sunshade net shading cooling moisture-retaining in green house.
Described transplanting process comprises the every day of blade face water spray sooner or later, cultivates 3 ~ 5 weeks, sees young leaves and start growth, show that the gold leaf Acer negundo. L of transplanting survives.
The present invention has following beneficial effect:
(1) in the present invention, the healthy and strong aseptic plantlet in vitro of a strain can cut into 3 ~ 6 length of tape rhizome sections, and reproduction coefficient is high, and band rhizome section just can grow up to whole plant after 2 ~ 3 weeks, and the breeding cycle is shorter, is a kind of effective way of gold leaf Acer negundo. L tissue-culturing rapid propagation.
(2) the Primary culture base used in the present invention, Initial culture base and squamous subculture are same medium, namely a kind of medium is only needed in the present invention, same medium can carry out breeding and culture of rootage simultaneously, cultivate simple flow, operate simple and easy, time saving and energy saving, easier can obtain gold leaf Acer negundo. L aseptic seedling.
(3) the gold leaf Acer negundo. L aseptic seedling after taking root in the present invention, robust growth, does not need special hardening, just can directly transplant, and shortens the time obtaining gold leaf Acer negundo. L.
(4) the gold leaf Acer negundo. L aseptic seedling obtained in the present invention is directly transplanted in green house, and survival rate is high, can reach 100%.
Accompanying drawing explanation
Fig. 1 is axillary bud sprouting in gold leaf Acer negundo. L Primary culture of the present invention;
Fig. 2 is the aseptic plantlet in vitro of gold leaf Acer negundo. L in gold leaf Acer negundo. L Initial culture of the present invention;
Fig. 3 is gold leaf Acer negundo. L squamous subculture situation of the present invention, wherein, and a: initial stage, b: later stage, c: culture of rootage, d: root system;
Fig. 4 is gold leaf Acer negundo. L transplant survival plant of the present invention.
Embodiment
Below in conjunction with embodiment, the present invention will be further described, and protection scope of the present invention is not limited to the following stated.
embodiment 1:a tissue culture and rapid propagation method for gold leaf Acer negundo. L, it comprises the following steps:
(1) explant sterilization: gather without the gold leaf Acer negundo. L tender stem segments of damage by disease and insect, remove blade, stem section is cut into 2cm stem with bud, on superclean bench, with alcohol sterilizing 30s, mercuric chloride sterilizing 5min, with aseptic water washing 4 times;
(2) Primary culture: be inoculated into by the stem with bud after sterilizing in Primary culture base, cultivates 2 weeks, axillalry bud lengthen by 2 cm in culturing room;
(3) Initial culture: axillalry bud is cut, be inoculated in Initial culture base, cultivate 3 weeks in culturing room, obtain the healthy and strong aseptic plantlet in vitro of gold leaf Acer negundo. L of taking root, after cultivating 3 weeks, plant strain growth 6cm, and take root, well developed root system, number of taking root is 2, the long 3cm of root, regrow out at axil place axillalry bud, and growth coefficient is 3;
(4) squamous subculture: with the aseptic plantlet in vitro of gold leaf Acer negundo. L of stalwartness for female parent, by segmentation, its stem section is cut into 3 length of tape leaf stem sections, is inoculated in subculture medium, carry out subculture, carry out culture of rootage simultaneously, cultivates 2 weeks, obtains gold leaf Acer negundo. L aseptic seedling,
(5) transplant: wash the medium on taking root gold leaf Acer negundo. L aseptic seedling strong off, directly be transplanted on the seedbed with the green house of commercially available sunshade net shading cooling moisture-retaining, controlling canopy temperature is 20 DEG C, wherein, in transplanting process, matrix used is perlite and humus mixing, and both weight ratios are 1:1.5.
In described step (2), step (3) and step (4), medium is 1/2DKW+0.1mg/LNAA+0.05mg/LIAA+ sucrose 30g/L+ agar 5g/L, pH5.8.
Cultivate indoor conditions in described step (2), step (3) and step (4) to be: intensity of illumination 2600LX, light application time 18h, temperature 22 DEG C.
Described transplanting process comprises the every day of blade face water spray sooner or later, cultivates 3 weeks, sees young leaves and start growth, show that the gold leaf Acer negundo. L of transplanting survives, and after 2 weeks, the survival rate of investigation gold leaf Acer negundo. L, survival rate was 100%.
embodiment 2:a tissue culture and rapid propagation method for gold leaf Acer negundo. L, it comprises the following steps:
(1) explant sterilization: gather without the gold leaf Acer negundo. L tender stem segments of damage by disease and insect, remove blade, stem section is cut into 3cm stem with bud, on superclean bench, with alcohol sterilizing 60s, mercuric chloride sterilizing 8min, with aseptic water washing 6 times;
(2) Primary culture: be inoculated into by the stem with bud after sterilizing in Primary culture base, cultivates 3 weeks in culturing room, and axillalry bud extends 4cm;
(3) Initial culture: axillalry bud is cut, be inoculated in Initial culture base, cultivate 2 weeks in culturing room, obtain the healthy and strong aseptic plantlet in vitro of gold leaf Acer negundo. L of taking root, after cultivating 4 weeks, plant strain growth 7cm, and take root, well developed root system, number of taking root is 12, the long 5cm of root, regrow out at axil place axillalry bud, and growth coefficient is 6;
(4) squamous subculture: with the aseptic plantlet in vitro of gold leaf Acer negundo. L of stalwartness for female parent, by segmentation, its stem section is cut into 6 length of tape leaf stem sections, be inoculated in subculture medium, carry out subculture, carry out culture of rootage simultaneously, cultivate 3 weeks, obtain gold leaf Acer negundo. L aseptic seedling;
(5) transplant: wash the medium on taking root gold leaf Acer negundo. L aseptic seedling strong off, directly be transplanted on the seedbed with the green house of commercially available sunshade net shading cooling moisture-retaining, controlling canopy temperature is 30 DEG C, wherein, in transplanting process, matrix used is perlite and humus mixing, and both weight ratios are 1:2.5.
In described step (2), step (3) and step (4), medium is 1/2DKW+0.1mg/LNAA+0.05mg/LIAA+ sucrose 30g/L+ agar 5g/L, pH6.3.
Cultivate indoor conditions in described step (2), step (3) and step (4) to be: intensity of illumination 3000LX, light application time 14h, temperature 26 DEG C.
Described transplanting process comprises the every day of blade face water spray sooner or later, cultivates 5 weeks, sees young leaves and start growth, show that the gold leaf Acer negundo. L of transplanting survives, and after 2 weeks, the survival rate of investigation gold leaf Acer negundo. L, survival rate was 100%.
embodiment 3:a tissue culture and rapid propagation method for gold leaf Acer negundo. L, it comprises the following steps:
(1) explant sterilization: gather without the gold leaf Acer negundo. L tender stem segments of damage by disease and insect, remove blade, stem section is cut into 2.3cm stem with bud, on superclean bench, with alcohol sterilizing 40s, mercuric chloride sterilizing 6min, with aseptic water washing 5 times;
(2) Primary culture: be inoculated into by the stem with bud after sterilizing in Primary culture base, cultivates 16 days in culturing room, and axillalry bud extends 2.8cm;
(3) Initial culture: axillalry bud is cut, be inoculated in Initial culture base, cultivate 16 days in culturing room, obtain the healthy and strong aseptic plantlet in vitro of gold leaf Acer negundo. L of taking root, cultivate after 23 days, plant strain growth 6.3cm, and take root, well developed root system, number of taking root is 6, the long 3.5cm of root, regrow out at axil place axillalry bud, and growth coefficient is 4;
(4) squamous subculture: with the aseptic plantlet in vitro of gold leaf Acer negundo. L of stalwartness for female parent, by segmentation, its stem section is cut into 4 length of tape leaf stem sections, be inoculated in subculture medium, carry out subculture, carry out culture of rootage simultaneously, cultivate 17 days, obtain gold leaf Acer negundo. L aseptic seedling;
(5) transplant: wash the medium on taking root gold leaf Acer negundo. L aseptic seedling strong off, directly be transplanted on the seedbed with the green house of commercially available sunshade net shading cooling moisture-retaining, controlling canopy temperature is 24 DEG C, wherein, in transplanting process, matrix used is perlite and humus mixing, and both weight ratios are 1:1.8.
In described step (2), step (3) and step (4), medium is 1/2DKW+0.1mg/LNAA+0.05mg/LIAA+ sucrose 30g/L+ agar 5g/L, pH6.0.
Cultivate indoor conditions in described step (2), step (3) and step (4) to be: intensity of illumination 2700LX, light application time 16h, temperature 24 DEG C.
Described transplanting process comprises the every day of blade face water spray sooner or later, cultivates 4 weeks, sees young leaves and start growth, show that the gold leaf Acer negundo. L of transplanting survives, and after 2 weeks, the survival rate of investigation gold leaf Acer negundo. L, survival rate was 99.5%.
embodiment 4:a tissue culture and rapid propagation method for gold leaf Acer negundo. L, it comprises the following steps:
(1) explant sterilization: gather without the gold leaf Acer negundo. L tender stem segments of damage by disease and insect, remove blade, stem section is cut into 2.7cm stem with bud, on superclean bench, with alcohol sterilizing 50s, mercuric chloride sterilizing 7min, with aseptic water washing 5 times;
(2) Primary culture: be inoculated into by the stem with bud after sterilizing in Primary culture base, cultivates 19 days in culturing room, and axillalry bud extends 3.5cm;
(3) Initial culture: axillalry bud is cut, be inoculated in Initial culture base, cultivate 19 days in culturing room, obtain the healthy and strong aseptic plantlet in vitro of gold leaf Acer negundo. L of taking root, cultivate after 26 days, plant strain growth 6.7cm, and take root, well developed root system, number of taking root is 9, the long 4.2cm of root, regrow out at axil place axillalry bud, and growth coefficient is 5;
(4) squamous subculture: with the aseptic plantlet in vitro of gold leaf Acer negundo. L of stalwartness for female parent, by segmentation, its stem section is cut into 5 length of tape leaf stem sections, be inoculated in subculture medium, carry out subculture, carry out culture of rootage simultaneously, cultivate 20 days, obtain gold leaf Acer negundo. L aseptic seedling;
(5) transplant: wash the medium on taking root gold leaf Acer negundo. L aseptic seedling strong off, directly be transplanted on the seedbed with the green house of commercially available sunshade net shading cooling moisture-retaining, controlling canopy temperature is 27 DEG C, wherein, in transplanting process, matrix used is perlite and humus mixing, and both weight ratios are 1:2.2.
In described step (2), step (3) and step (4), medium is 1/2DKW+0.1mg/LNAA+0.05mg/LIAA+ sucrose 30g/L+ agar 5g/L, pH6.2.
Cultivate indoor conditions in described step (2), step (3) and step (4) to be: intensity of illumination 2800LX, light application time 17h, temperature 25 DEG C.
In described Initial culture, axillalry bud is in Initial culture base, and cultivate 26 days in culturing room, plant strain growth 6.7cm, and take root, well developed root system, number of taking root is 9, the long 4.2cm of root, and regrow out at axil place axillalry bud, and growth coefficient is 5.
Described transplanting process comprises the every day of blade face water spray sooner or later, cultivates 4 weeks, sees young leaves and start growth, show that the gold leaf Acer negundo. L of transplanting survives, and after 2 weeks, the survival rate of investigation gold leaf Acer negundo. L, survival rate was 100%.
Draw from embodiment 1 ~ 4: the present invention utilizes tissue culture technique, medium used is same, and the gold leaf Acer negundo. L reproduction coefficient of acquisition is 3 ~ 6, and group is trained the gold leaf Acer negundo. L obtained and is directly transplanted in green house, survival rate is up to 100%.
Claims (3)
1. a tissue culture and rapid propagation method for gold leaf Acer negundo. L, is characterized in that, it comprises the following steps:
(1) explant sterilization: gather without the gold leaf Acer negundo. L tender stem segments of damage by disease and insect, remove blade, stem section is cut into 2 ~ 3cm stem with bud, on superclean bench, with alcohol sterilizing 30 ~ 60s, mercuric chloride sterilizing 5 ~ 8min, with aseptic water washing 4 ~ 6 times;
(2) Primary culture: be inoculated into by the stem with bud after sterilizing in Primary culture base, cultivates 2 ~ 3 weeks in culturing room, and axillalry bud extends 2 ~ 4cm; Described Primary culture base is 1/2DKW+0.1mg/LNAA+0.05mg/LIAA+ sucrose 30g/L+ agar 5g/L, pH5.8 ~ 6.3;
(3) Initial culture: axillalry bud is cut, be inoculated in Initial culture base, cultivate 2 ~ 3 weeks in culturing room, obtain the healthy and strong aseptic plantlet in vitro of gold leaf Acer negundo. L of taking root, cultivate after 3 ~ 4 weeks, plant strain growth 6 ~ 7cm, number of taking root is 2 ~ 12, long 3 ~ the 5cm of root, regrow out at axil place axillalry bud, growth coefficient 3 ~ 6; Described Initial culture base is 1/2DKW+0.1mg/LNAA+0.05mg/LIAA+ sucrose 30g/L+ agar 5g/L, pH5.8 ~ 6.3;
(4) squamous subculture: with the aseptic plantlet in vitro of gold leaf Acer negundo. L of stalwartness for female parent, its stem section is cut into 3 ~ 6 length of tape leaf stem sections, is inoculated in subculture medium and cultivates 2 ~ 3 weeks, obtain the whole plant of gold leaf Acer negundo. L aseptic seedling; Described subculture medium is 1/2DKW+0.1mg/LNAA+0.05mg/LIAA+ sucrose 30g/L+ agar 5g/L, pH5.8 ~ 6.3;
(5) transplant: wash the medium on taking root gold leaf Acer negundo. L aseptic seedling strong off, be directly transplanted on green house seedbed, controlling canopy temperature is 20 ~ 30 DEG C.
2. the tissue culture and rapid propagation method of a kind of gold leaf Acer negundo. L according to claim 1, it is characterized in that, condition of culture in step (2), step (3) and step (4) is: intensity of illumination 2600LX ~ 3000LX, light application time 14 ~ 18h, temperature 22 ~ 26 DEG C.
3. the tissue culture and rapid propagation method of a kind of gold leaf Acer negundo. L according to claim 1, is characterized in that, in described transplanting process, transplanting medium is perlite and humus mixing, and both weight ratios are 1:1.5 ~ 2.5.
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| CN104920225B (en) * | 2015-07-03 | 2017-10-24 | 西北农林科技大学 | A kind of tissue culture and rapid propagation method of gold leaf Acer negundo. L |
| CN105532459B (en) * | 2015-12-30 | 2017-10-20 | 四川七彩林业开发有限公司 | A kind of tissue culture and rapid propagation method of Acer palmatum orange dream |
| CN105638464B (en) * | 2015-12-30 | 2018-01-09 | 四川七彩林业开发有限公司 | A kind of tissue culture and rapid propagation method of silk ribbon grass |
| CN105940920A (en) * | 2016-05-20 | 2016-09-21 | 芜湖欧标农业发展有限公司 | Seedling growing method for hard wood cutting of Acer negundo Linn in North America |
| CN107455261B (en) * | 2017-09-15 | 2019-08-13 | 安徽农业大学 | A kind of method of Acer saccharum tissue culture quick breeding |
| CN110692519B (en) * | 2019-11-11 | 2021-04-02 | 四川七彩林科股份有限公司 | Tissue culture and rapid propagation method for pseudoforsythia aureocauda |
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