CN101292630A - Rieger Begonia tissue culture quick replication method - Google Patents
Rieger Begonia tissue culture quick replication method Download PDFInfo
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- CN101292630A CN101292630A CNA2008100678796A CN200810067879A CN101292630A CN 101292630 A CN101292630 A CN 101292630A CN A2008100678796 A CNA2008100678796 A CN A2008100678796A CN 200810067879 A CN200810067879 A CN 200810067879A CN 101292630 A CN101292630 A CN 101292630A
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Abstract
The invention relates to a rapid propagation method for culturing Rieger begonias tissue, which comprises the following steps that: explant material is treated, that is, the explant material is selected for treatment to prepare the explant meeting the requirement; then inducement is carried out, that is, the explant prepared in the first step gets vaccinated to culture medium to induce bud on inducing conditions; the inducing condition that the light intensity with illuminance is less than 1500lx is met. As the lighting condition during the fast propagation of Rieger begonias tissue is further optimized, the application of the rapid propagation method for culturing Rieger begonias tissue causes the inducement time to be shorter and ensures the inducement rate to be higher and reach 100 percent.
Description
Technical field
The invention belongs to technical field of bioengineering, relate to plant tissue culture technique, be specifically related to a kind of Rieger Begonia tissue culture quick replication method.
Background technology
Rieger Begonia (Rieger begonias) claims the rose Malus spectabilis again, and the perennial herb flowers of Begoniaceae Begonia are hybridization strains of napiform root Malus spectabilis and wild begonia, belong to high-grade potted flower.Generally can't form seed, all adopt cottage propagation mostly, but Rieger Begonia is relatively stricter to temperature requirement, To Be Protected from Heat is afraid of cold, the unsuitable strong illumination of most kinds, and the cuttage maintenance is not easy very much, and the degeneration of Rieger Begonia germ plasm resource is comparatively serious, and the quality of cottage propagation is very undesirable.The technology of tissue culture propagating plant is a kind of special mode of nourishing and generating, and is commonly referred to as quick propagating technology or micro-propagation technique now.It is under aseptic condition, utilizes the part of plant corpus, comprises cell, tissue and organ, the method for breeding plant under manually operated nutrition and environmental condition.This technology comparative maturity, equipment is fairly simple, and specification requirement is very not high, and economic benefit is considerable, more and more is subject to people's attention.Therefore, adopt the batch production mode of production to breed the Rieger Begonia seedling and can prevent deterioration of variety, guarantee seedling quality, the seedling of a large amount of high-quality cheapnesss can be provided again.Prior art is generally got tender blade, stem section or the stem apex of Rieger Begonia children and is made explant, with MS is minimal medium, regulates the basic element of cell division and growth hormone ratio, in 25 to 28 degrees centigrade of culturing room, under 14 to 16 hours illumination conditions, can induce in about 40 to 60 days to differentiate seedling.But the inductivity of emerging of prior art is not high; And induction time is also longer, can not adapt to the market demand more fast.
Summary of the invention
Technical problem to be solved by this invention provides and a kind ofly can breed the Rieger Begonia seedling sooner, and the higher Rieger Begonia tissue culture quick replication method of inductivity.
For solving the problems of the technologies described above, the present invention adopts following technical scheme:
Rieger Begonia tissue culture quick replication method comprises the steps:
S1), explant material is handled: promptly select for use explant material to handle, prepare satisfactory explant;
S2), induce: be about to the explant described in the step S1 and be inoculated on the medium, under inductive condition, induce and sprout; Described inductive condition comprises the low light level photograph condition of illuminance less than 1500lx.
Further, the low light level described in the step S2 comprises two stages according to condition, and the illuminance in first stage is 100～200lx, and the illuminance in second stage is 1100～1200lx.Every day, light application time was 12 hours.
The light application time in described first stage is 10～14 days.
The light application time in described second stage is 18～28 days.
Further, the prescription of medium described in the step S2 is MS+6-benzyl aminopterin-induced syndrome 0.4～0.6mg/l+ methyl 0.1～0.3mg/l+ sucrose 25～30g+ agar 6～8g.
In the optimized technical scheme, described step S1 explant material is handled and is comprised cleaning, sterilization and cutting; Described cleaning will be used to make the explant material of explant and clean; Described sterilization is disinfected the explant material of cleaning; Explant material after described cutting will be sterilized is cut into the explant of suitable size.
Described explant material is the young leaflet tablet of Rieger Begonia.
The water logging that described cleaning comprises running water flushing and contains liquid detergent is washed.
Described sterilization comprises that the liquor natrii hypochloritis of alcohol disinfecting and adding Tween-20 embathes sterilization.
Described being cut to removed the vanes block that is cut into 0.5～1cm * 0.5～1cm size after leaf margin and the petiole with explant material.
Adopt the Rieger Begonia tissue culture quick replication method of technical solution of the present invention, the beneficial effect that is compared with the prior art is:
Owing to further optimized the illumination condition in the Rieger Begonia tissue-culturing quick-propagation process, made that the induction time of Rieger Begonia is shorter; And inductivity is higher, and inductivity can reach 100%.
Because the low light level is further optimized two stages that are decomposed into according to condition, and more accurate in the illuminance control in two stages, can reach and better induce effect.
The light application time of shining condition owing to the low light level with first stage is defined as 10～14 days, and process control is more accurate, better effects if.
The light application time of shining condition owing to the low light level with second stage is defined as 18～28 days, and process control is more accurate, better effects if.
Because adopt MS+6-benzyl aminopterin-induced syndrome 0.4～0.6mg/l+ methyl 0.1～0.3mg/l+ sucrose 25～30g+ agar 6～8g as medium, the inducing culture effect is better.
Comprise cleaning, sterilization and cutting because explant material is handled, can obtain the explant of high-quality more, for quick breeding method for tissue culture lays a good foundation.
Because the young leaflet tablet of preferred Rieger Begonia as explant material, guarantees for tissue-culturing quick-propagation provides good material.
Wash owing to clean the water logging that comprises the running water flushing and contain liquid detergent, can be so that cleaning be more thorough.
Because sterilization comprises the liquor natrii hypochloritis of alcohol disinfecting and adding Tween-20 and embathes sterilization that it is more thorough to sterilize.
Since with explant material remove be cut into 0.5～1cm * 0.5～1cm size after leaf margin and the petiole vanes block as explant, inducing culture is convenient, and better effects if.
Embodiment
Below in conjunction with instantiation enforcement of the present invention is described further:
Example one
1, explant material is handled: comprise to explant material clean, processing such as sterilization and cutting, to prepare the explant that meets the requirements.Be specially: get the young leaflet tablet of the Rieger Begonia of red flower,, wash 20～30min with the water logging that adds liquid detergent again with washing 20～30min in the running water.Disinfection on superclean bench after cleaning up, earlier soak 20s with 70% alcohol, embathe 10min in clorox (available chlorine 〉=7.5%) solution with 10% (V/V) that add Tween-20 again, it is standby to put into aseptic bottle after the blade sterilization is finished.When needing blade is taken out from aseptic bottle, on superclean bench, removes after the leaf margin and petiole of blade, be cut into then 0.5～1cm * 0.5～1cm size vanes block explant.
2, induce and sprout: will be seeded in prescription as the vanes block back side of explant down on the medium of MS+6-benzyl aminopterin-induced syndrome 0.4mg/l+ methyl 0.1mg/l+ sucrose 30g+ agar 6g.Be placed on earlier under 100～200lx illumination and cultivated 14 days, move on to again under 1100～1200lx illumination and cultivated 18 days.Illumination every day 12 hours, cultivation temperature are 22～24 degrees centigrade, below identical therewith.Find inductivity up to 100% through statistics, and induction time only is 32 days.
Annotate: grow thickly explant number/total explant number * 100% of bud of inductivity (%)=occur.6-benzyl aminopterin-induced syndrome: 6-BA (6-Benzyl Aminopurine); Methyl: NAA (naphtalene acetic acid), below identical.
Example two
Get the young leaflet tablet of the Rieger Begonia of pink flower, obtain explant to handle with example one identical method.Then explant is seeded on the medium identical with embodiment one, is placed on earlier under 100～200lx illumination and cultivated 14 days, move on to cultivation 1100～1200lx illumination under then, the inductivity that 18 days metaplexus sprout reaches 99%, 21 day metaplexus inductivity of sprouting and reaches 100%.
Example three
Get the young leaflet tablet of the Rieger Begonia of red flower, obtain explant to handle with example one identical method.Then explant is seeded in prescription on the medium of MS+6-benzyl aminopterin-induced syndrome 0.6mg/l+ methyl 0.3mg/l+ sucrose 30g+ agar 6g, under 100～200lx illumination condition, cultivated 14 days earlier, move on to then under 1100～1200lx illumination condition and cultivate, the inductivity that 18 days metaplexus sprout reaches the inductivity that 90%, 28 day metaplexus sprouts and reaches 100%.
Example four
Get the young leaflet tablet of the Rieger Begonia of red flower, obtain explant to handle with example one identical method.Then explant is seeded in prescription on the medium of MS+6-benzyl aminopterin-induced syndrome 0.4mg/l+ methyl 0.1mg/l+ sucrose 30g+ agar 6g.Cultivated 10 days under 100～200lx illumination condition earlier, move on to cultivation 1100～1200lx illumination under then, the inductivity that 22 days metaplexus sprout reaches the inductivity that 97%, 27 day metaplexus sprouts and reaches 100%.
Example five
Get the young leaflet tablet of the Rieger Begonia of red flower, obtain explant to handle with example one identical method.Then explant is seeded in prescription on the medium of MS+6-benzyl aminopterin-induced syndrome 0.6mg/l+ methyl 0.1mg/l+ sucrose 25g+ agar 8g.Cultivated 14 days under 100～200lx illumination condition earlier, move on to cultivation 1100～1200lx illumination under then, the inductivity that 22 days metaplexus sprout reaches the inductivity that 87%, 29 day metaplexus sprouts and reaches 100%.
Illumination condition is one of essential condition of tissue culture.Key technology of the present invention is, on suitable hormone concentration basis, under low light condition less than 1500lx, by change illumination condition improve Rieger Begonia grow thickly bud inductivity and shorten induction time.
Above content be in conjunction with concrete preferred implementation to further describing that the present invention did, can not assert that concrete enforcement of the present invention is confined to these explanations.For the general technical staff of the technical field of the invention, without departing from the inventive concept of the premise, can also make some simple deduction or replace, all should be considered as belonging to protection scope of the present invention.
Claims (10)
1, Rieger Begonia tissue culture quick replication method comprises the steps:
S1), explant material is handled: promptly select for use explant material to handle, prepare satisfactory explant;
S2), induce: be about to the explant described in the step S1 and be inoculated on the medium, under inductive condition, induce and sprout; Described inductive condition comprises the low light level photograph condition of illuminance less than 1500lx.
2, Rieger Begonia tissue culture quick replication method as claimed in claim 1, it is characterized in that, the low light level described in the step S2 comprises two stages according to condition, and the illuminance in first stage is 100～200lx, and the illuminance in second stage is 1100～1200lx.
3, Rieger Begonia tissue culture quick replication method as claimed in claim 2 is characterized in that, the light application time in described first stage is 10～14 days.
4, Rieger Begonia tissue culture quick replication method as claimed in claim 2 is characterized in that, the light application time in described second stage is 18～28 days.
5, Rieger Begonia tissue culture quick replication method as claimed in claim 1 is characterized in that, the prescription of medium described in the step S2 is MS+6-benzyl aminopterin-induced syndrome 0.4～0.6mg/l+ methyl 0.1～0.3mg/l+ sucrose 25～30g+ agar 6～8g.
As any described Rieger Begonia tissue culture quick replication method in the claim 1 to 5, it is characterized in that 6, described step S1 explant material is handled and comprised cleaning, sterilization and cutting; Described cleaning will be used to make the explant material of explant and clean; Described sterilization is disinfected the explant material of cleaning; Explant material after described cutting will be sterilized is cut into the explant of suitable size.
7, Rieger Begonia tissue culture quick replication method as claimed in claim 6 is characterized in that, described explant material is the young leaflet tablet of Rieger Begonia.
8, Rieger Begonia tissue culture quick replication method as claimed in claim 6 is characterized in that, the water logging that described cleaning comprises running water flushing and contains liquid detergent is washed.
9, Rieger Begonia tissue culture quick replication method as claimed in claim 6 is characterized in that, described sterilization comprises that the liquor natrii hypochloritis of alcohol disinfecting and adding Tween-20 embathes sterilization.
10, Rieger Begonia tissue culture quick replication method as claimed in claim 6 is characterized in that, described being cut to removed the vanes block that is cut into 0.5～1cm * 0.5～1cm size after leaf margin and the petiole with explant material.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2008100678796A CN101292630B (en) | 2008-06-18 | 2008-06-18 | Rieger Begonia tissue culture quick replication method |
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| CN2008100678796A CN101292630B (en) | 2008-06-18 | 2008-06-18 | Rieger Begonia tissue culture quick replication method |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102217549A (en) * | 2011-06-03 | 2011-10-19 | 张曦予 | Culture method of rieger begonias test tube flowers |
| CN102696487A (en) * | 2012-07-05 | 2012-10-03 | 江苏省中国科学院植物研究所 | Method for building leaf in vitro regeneration system of begonia rex |
| CN103155866A (en) * | 2011-12-10 | 2013-06-19 | 天水市果树研究所 | Malus zumi tissue culture rapid propagation seedling raising method |
| CN103202228A (en) * | 2013-04-01 | 2013-07-17 | 肇庆学院 | One-step seedling and efficient in-vitro propagation method with gynura bicolor leaves |
| CN105230455A (en) * | 2015-10-11 | 2016-01-13 | 云南省农业科学院花卉研究所 | Method for cultivating begonia fangii using water |
| CN106212274A (en) * | 2016-07-15 | 2016-12-14 | 武汉市农业科学技术研究院林业果树科学研究所 | The method for tissue culture of special beautiful Caulis et folium euphorbiae milii |
| CN110476810A (en) * | 2019-08-20 | 2019-11-22 | 广州市名卉景观科技发展有限公司 | A kind of method of Rieger Begonia fast breeding |
| CN112273237A (en) * | 2020-11-23 | 2021-01-29 | 山西省农业科学院园艺研究所 | Tissue culture and rapid propagation method for Rieger begonia |
| CN115024223A (en) * | 2022-06-27 | 2022-09-09 | 安徽农业大学 | In-vitro preservation method for malus spectabilis germplasm |
-
2008
- 2008-06-18 CN CN2008100678796A patent/CN101292630B/en not_active Expired - Fee Related
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102217549A (en) * | 2011-06-03 | 2011-10-19 | 张曦予 | Culture method of rieger begonias test tube flowers |
| CN102217549B (en) * | 2011-06-03 | 2012-07-25 | 张曦予 | Culture method of rieger begonias test tube flowers |
| CN103155866B (en) * | 2011-12-10 | 2015-06-17 | 天水市果树研究所 | Malus zumi tissue culture rapid propagation seedling raising method |
| CN103155866A (en) * | 2011-12-10 | 2013-06-19 | 天水市果树研究所 | Malus zumi tissue culture rapid propagation seedling raising method |
| CN102696487A (en) * | 2012-07-05 | 2012-10-03 | 江苏省中国科学院植物研究所 | Method for building leaf in vitro regeneration system of begonia rex |
| CN103202228A (en) * | 2013-04-01 | 2013-07-17 | 肇庆学院 | One-step seedling and efficient in-vitro propagation method with gynura bicolor leaves |
| CN103202228B (en) * | 2013-04-01 | 2014-08-27 | 肇庆学院 | One-step seedling and efficient in-vitro propagation method with gynura bicolor leaves |
| CN105230455A (en) * | 2015-10-11 | 2016-01-13 | 云南省农业科学院花卉研究所 | Method for cultivating begonia fangii using water |
| CN105230455B (en) * | 2015-10-11 | 2018-05-04 | 云南省农业科学院花卉研究所 | Fang Shi begonia water planting propagation methods |
| CN106212274A (en) * | 2016-07-15 | 2016-12-14 | 武汉市农业科学技术研究院林业果树科学研究所 | The method for tissue culture of special beautiful Caulis et folium euphorbiae milii |
| CN110476810A (en) * | 2019-08-20 | 2019-11-22 | 广州市名卉景观科技发展有限公司 | A kind of method of Rieger Begonia fast breeding |
| CN112273237A (en) * | 2020-11-23 | 2021-01-29 | 山西省农业科学院园艺研究所 | Tissue culture and rapid propagation method for Rieger begonia |
| CN115024223A (en) * | 2022-06-27 | 2022-09-09 | 安徽农业大学 | In-vitro preservation method for malus spectabilis germplasm |
| CN115024223B (en) * | 2022-06-27 | 2023-03-14 | 安徽农业大学 | In-vitro preservation method for malus spectabilis germplasm |
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| CN101292630B (en) | 2011-11-02 |
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