CN102283130A - Method for culturing and fast propagating tissue of chimaera hosta plantaginea plant - Google Patents
Method for culturing and fast propagating tissue of chimaera hosta plantaginea plant Download PDFInfo
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- CN102283130A CN102283130A CN2011102068791A CN201110206879A CN102283130A CN 102283130 A CN102283130 A CN 102283130A CN 2011102068791 A CN2011102068791 A CN 2011102068791A CN 201110206879 A CN201110206879 A CN 201110206879A CN 102283130 A CN102283130 A CN 102283130A
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Abstract
The invention provides a method for culturing and fast propagating a tissue of a chimaera hosta plantaginea plant, which utilizes a plumule of the chimaera hosta plantaginea as an explant to perform tissue culturing and fast propagation and comprises the following steps of disinfecting, primary culturing, subculture and rooting culturing. The method has a high propagation coefficient, is suitable for commercial production and fills in the blank of requirements of the market on the chimaera hosta plantaginea. In the invention, the plumule of the chimaera hosta plantaginea is used as the explant, and the plant regeneration and the fast propagation technologies of fragrant plantain lily flower are researched. A large number of requirements of commercial production can be met through an overall industrialization process of plant regeneration, propagation, taking root and seedling and high culture medium multiplication coefficient.
Description
Technical field
The present invention relates to the tissue culture and rapid propagation method of a kind of chimera jade hairpin plant.
Background technology
Chimera is to grow same individuality or the organ that forms jointly by the cell of different genetic character, and cell mutation is the basis that chimera forms.Sudden change all can take place in any plant.Plant chimeric body (plant chimera) is meant and contains genotypical plant tissue or plant individual more than 2 kinds or 2 kinds.Baur (1909) has carried out detailed research to the green core pattern geranium in yellow limit (Pelargonium zonale) plant, find its growing point original hase (shoot apical meristem, SAM), stem stalk, petiole and leaf form regularly by 2 kinds of genotype cells, is referred to as the plant chimeric body then.On this basis, formed the basic theory of plant chimeric body after further study, for the application of plant chimeric body and the breeding of exotic flowers and rare herbs provide foundation.
Jade hairpin (
Hosta plantigineaAscherson), having another name called beautiful spring rod, white crane celestial being, white calyx, is a kind of good shade plant, not only considerable leaf but also considerable flower, simultaneously, the jade hairpin complete stool all can be used as medicine, flower be used as medicine have dampness removing, the merit of adjusting meridian and stopping leukorrhea, root be used as medicine have clearing heat for detumescence, the merit of detoxifcation pain relieving, leaf can removing toxicity for detumescence.The embedding of jade hairpin and body comprise green core, green handle greenery, bottle green, blade is green entirely, Da Lan spends, 7 types of the yellow core of Xiao Lan flower and green limit.
Aspect breeding practice, it is numerous to utilize plant division etc. to induce Different Organs to obtain the stable expansion of chimera plant.At present, jade hairpin is many breeds with offshoot, and reproduction coefficient is low, and the cycle is long, can not satisfy the needs in production and market.The tissue-culturing rapid propagation of research jade hairpin can satisfy the wilderness demand of commercially producing.
Summary of the invention
The object of the present invention is to provide the tissue culture and rapid propagation method of a kind of chimera jade hairpin plant, this method reproduction coefficient height is fit to commercially produce, and has filled up market to chimera jade hairpin demand blank.
The tissue culture and rapid propagation method of a kind of chimera jade hairpin plant of the present invention comprises sterilization, initial culture, successive transfer culture, culture of rootage, utilizes the young shoot of chimera jade hairpin to carry out tissue culture and breeding fast for explant.
Described disinfectant method is: young shoot is scrubbed gently with washing powder water logging bubble 10-15min and with fur earlier, again with the flushing with clean water 30-45min that flows, 75% alcohol-pickled 30-45s, 0.1% mercuric chloride soaks 10-15min, constantly stirs, and uses aseptic water washing 3-5 time again, blot with filter paper, be cut into 0.5-1cm
3Square, young shoot base portion forward inserts in the medium and carries out initial culture.
When making initial culture, be medium, add 1mg/L-3.0mg/L6-BA and 0.1mg/L-0.5mg/LNAA with MS with the jade hairpin young shoot.
When making successive transfer culture, be medium, add 0.5mg/L-2.0mg/L6-BA and 0.1mg/L-0.3mg/LNAA with MS with the bud that induces.
When carrying out expanding propagation, be medium, add 1mg/L-5mg/L6-BA and 0.1mg/L-0.5mg/LNAA with MS with a large amount of clumps of buds that induce.
During culture of rootage bud is forwarded to: 1/2MS-1/4MS medium adds the active carbon of 0.1mg/L-0.3mg/LNAA and 0.5mg/L-1.0mg/L IBA and 0.1%-0.2%; Described 1/2MS-1/4MS refers to the amount of macroelement.
When the root of test-tube plantlet grows to 2-3cm, be placed on room temperature lower refining seedling 1-2 week, be transplanted to then and be equipped with in leaf mould and the perlitic cave dish; Root system covers with the cave dish after one month, and beginning simultaneously has young leaves to grow, the seedling stalwartness, and growth potential is strong, and transplanting survival rate reaches more than 95%.
The present invention is an explant with chimera jade hairpin young shoot, has studied plant regeneration and the quick propagating technology of fragrant plantain lily flower.By the present invention finish plant regeneration, expand numerous, take root, whole industry process and the medium growth coefficient height of Cheng Miao can satisfy the wilderness demand of commercially producing.
Description of drawings
Fig. 1,2 is chimera jade hairpin initial culture figure as a result;
The bud of growing thickly that Fig. 3 ~ 5 obtain for enrichment culture;
Fig. 6 ~ 8 are the cultivation results figure of taking root;
Fig. 9 is factorial seedling growth figure;
Figure 10 ~ 12 are green limit yellow core chimera type transplant survival seedling.
Embodiment
Be specific embodiments of the present invention below, further describe the present invention, but the present invention be not limited only to this.
Embodiment 1
(1) test material: the length of getting stalwartness, no damage by disease and insect is that 2-3 centimetres of green limit yellow core chimera young shoots are as explant.
(2) acquisition of sterilizable material: earlier with washing gently about washing powder water logging bubble 10-15 min and with fur, again with the flushing with clean water 30-45 min that flows, 75% alcohol-pickled 30-45 s, 0.1% mercuric chloride soaks 10-15min, constantly stir, use aseptic water washing 3-5 time again, blot, be cut into 0.5-1cm with filter paper
3Square, young shoot base portion forward inserts in the medium and cultivates observation.
(3) medium: add in the medium of 6-BA3.0mg/L and NAA0.5mg/L than being easier to induce the bud of growing thickly at MS; 6-BA2.0mg/L and the combination of NAA0.3mg/L is the medium of optimum bud propagation; With 1/2MS is minimal medium, and the active carbon that adds 0.3mg/LNAA and 1.0mg/LIBA and 0.2% helps taking root of jade hairpin test-tube plantlet most.Simultaneously, pay special attention to prevent the pollution of endophyte, suitably adding the 2mL gentamycin solution in medium can be anti-pollution effectively in advance.
(4) condition of culture: every day intensity of illumination 1200-1500lx, lighting delay number 10-12h, temperature is 15 ± 2 ℃.
Rising along with 6-BA concentration, growth coefficient increases, illustrate that 6-BA has certain facilitation to the propagation of bud, but when the concentration of 6-BA is elevated to 5mg/L, growth coefficient but significantly descends, and the growth of bud seedling is short and small, and blade occurs aging, illustrates that the 6-BA of high concentration can suppress the propagation of bud on the contrary.Find that simultaneously vitrification phenomenon appears in the new bud of sprouting of minority stem section when NAA gets 1.0mg/L, and all do not have this phenomenon of generation when getting 0.1-0.5mg/L.Taking all factors into consideration the above-mentioned factor, when making initial culture with the jade hairpin young shoot, is medium with MS, adds 3.0mg/L6-BA and 0.5mg/LNAA, and the growth coefficient maximum reaches 95%, and the bud seedling grows fine by (table 1), as shown in Figure 1, 2.
The propagation situation of bud under different medium during table 1 initial culture
When making successive transfer culture with the bud that induces, cultivation effect is best under the MS medium that adds 2.0mg/L6-BA and 0.3mg/LNAA, and the bud that differentiates is many, and color is light green, stalwartness, and quality good (table 2) is shown in Fig. 3,4.
The propagation situation of bud under different medium during table 2 successive transfer culture
Discovery is observed in cultivation through 4 weeks, and IBA and active carbon can promote taking root of test-tube plantlet, but when the IBA excessive concentration, the test-tube plantlet base portion expands and rooting rate descends.It is thinner and more delicate not add in the medium of active carbon root, is not suitable for transplanting.Therefore, the active carbon that 1/2MS adds 0.3mg/LNAA and 1.0mg/LIBA and 0.2% is the optimal medium that promotes that jade hairpin is taken root, and the bud seedling rooting is many and sturdy, is fit to transplant (table 3), shown in Fig. 6-8.Figure 10 ~ 12 are green limit yellow core chimera type transplant survival seedling, and as seen it has kept chimeric characteristic well.
The different medium of table 3 are to the influence of jade hairpin rooting of vitro seedling
Claims (7)
1. the tissue culture and rapid propagation method of a chimera jade hairpin plant comprises sterilization, initial culture, successive transfer culture, culture of rootage, it is characterized in that: utilize the young shoot of chimera jade hairpin to carry out tissue culture and breeding fast for explant.
2. the tissue culture and rapid propagation method of chimera jade hairpin plant according to claim 1, it is characterized in that: described disinfectant method is: young shoot is scrubbed gently with washing powder water logging bubble 10-15min and with fur earlier, again with the flushing with clean water 30-45min that flows, 75% alcohol-pickled 30-45s, 0.1% mercuric chloride soaks 10-15min, constantly stirs, and uses aseptic water washing 3-5 time again, blot with filter paper, be cut into 0.5-1cm
3Square, young shoot base portion forward inserts in the medium and carries out initial culture.
3. the tissue culture and rapid propagation method of chimera jade hairpin plant according to claim 1 is characterized in that: when making initial culture with the jade hairpin young shoot, be medium with MS, add 1mg/L-3.0mg/L6-BA and 0.1mg/L-0.5mg/LNAA.
4. the tissue culture and rapid propagation method of chimera jade hairpin plant according to claim 1 is characterized in that: when making successive transfer culture with the bud that induces, be medium with MS, add 0.5mg/L-2.0mg/L6-BA and 0.1mg/L-0.3mg/LNAA.
5. the tissue culture and rapid propagation method of chimera jade hairpin plant according to claim 4 is characterized in that: when carrying out expanding propagation with a large amount of clumps of buds that induce, be medium with MS, add 1mg/L-5mg/L6-BA and 0.1mg/L-0.5mg/LNAA.
6. the tissue culture and rapid propagation method of chimera jade hairpin plant according to claim 1 is characterized in that: during culture of rootage bud is forwarded to: 1/2MS-1/4MS medium adds the active carbon of 0.1mg/L-0.3mg/LNAA and 0.5mg/L-1.0mg/L IBA and 0.1%-0.2%; Described 1/2MS-1/4MS refers to the amount of macroelement.
7. the tissue culture and rapid propagation method of chimera jade hairpin plant according to claim 1 is characterized in that: when the root of test-tube plantlet grows to 2-3cm, be placed on room temperature lower refining seedling 1-2 week, be transplanted to then and be equipped with in leaf mould and the perlitic cave dish; Root system covers with the cave dish after one month, and beginning simultaneously has young leaves to grow, the seedling stalwartness, and growth potential is strong, and transplanting survival rate reaches more than 95%.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102860261A (en) * | 2012-10-11 | 2013-01-09 | 山东鑫秋种业科技有限公司 | Method for preparing tissue culture medium of hosta undulata baileys |
| CN103858770A (en) * | 2014-04-03 | 2014-06-18 | 中国科学院植物研究所 | Rapid hosta plantagineu propagation method |
| CN103875535A (en) * | 2014-04-03 | 2014-06-25 | 中国科学院植物研究所 | Rapid proliferation method of hosta undulata bailey |
| CN104304035A (en) * | 2014-11-14 | 2015-01-28 | 黑龙江省科学院自然与生态研究所 | Tissue culture and rapid propagation method of plantain lily |
| CN104303745A (en) * | 2014-09-24 | 2015-01-28 | 江苏省林业科学研究院 | Fragrant plantain lily tissue culture seedling vitro-bottle rooting method |
| CN110972945A (en) * | 2019-12-19 | 2020-04-10 | 美尚生态景观股份有限公司 | Culture medium and culture method for tissue culture of plantain plantaginea |
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| CN1965643B (en) * | 2006-08-10 | 2011-04-06 | 沈阳市农业科学院 | Tissue culture method of Hosta plantiginea Ascherson |
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| CN1965643B (en) * | 2006-08-10 | 2011-04-06 | 沈阳市农业科学院 | Tissue culture method of Hosta plantiginea Ascherson |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102860261A (en) * | 2012-10-11 | 2013-01-09 | 山东鑫秋种业科技有限公司 | Method for preparing tissue culture medium of hosta undulata baileys |
| CN103858770A (en) * | 2014-04-03 | 2014-06-18 | 中国科学院植物研究所 | Rapid hosta plantagineu propagation method |
| CN103875535A (en) * | 2014-04-03 | 2014-06-25 | 中国科学院植物研究所 | Rapid proliferation method of hosta undulata bailey |
| CN103875535B (en) * | 2014-04-03 | 2015-09-02 | 中国科学院植物研究所 | A kind of method of floral leaf jade hairpin fast breeding |
| CN103858770B (en) * | 2014-04-03 | 2016-05-04 | 中国科学院植物研究所 | A kind of method of floral leaf jade hairpin Fast-propagation |
| CN104303745A (en) * | 2014-09-24 | 2015-01-28 | 江苏省林业科学研究院 | Fragrant plantain lily tissue culture seedling vitro-bottle rooting method |
| CN104304035A (en) * | 2014-11-14 | 2015-01-28 | 黑龙江省科学院自然与生态研究所 | Tissue culture and rapid propagation method of plantain lily |
| CN104304035B (en) * | 2014-11-14 | 2016-06-15 | 黑龙江省科学院自然与生态研究所 | The tissue cultivation rapid breeding method of golden-rimmed jade hairpin |
| CN110972945A (en) * | 2019-12-19 | 2020-04-10 | 美尚生态景观股份有限公司 | Culture medium and culture method for tissue culture of plantain plantaginea |
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