CN103704135A - In-vitro rapid propagation method for plantains - Google Patents
In-vitro rapid propagation method for plantains Download PDFInfo
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- CN103704135A CN103704135A CN201310710565.4A CN201310710565A CN103704135A CN 103704135 A CN103704135 A CN 103704135A CN 201310710565 A CN201310710565 A CN 201310710565A CN 103704135 A CN103704135 A CN 103704135A
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Abstract
The invention provides an in-vitro rapid propagation method for plantains. The in-vitro rapid propagation method for the plantains comprises the following steps: (1) callus induction culture, namely cutting off leaf veins and leaf margins of sterilized and disinfected plantains and cutting into small blocks; inoculating the plantains to a callus induction culture medium; and carrying out regular culture to form calluses and adventitious buds; and (2) rooting culture, namely selecting the adventitious buds, with the bud lengths of not more than 2cm-3cm, obtained in the step (1); cutting off the adventitious buds from the calluses and continuing to carry out callus induction culture on the residual calluses and the adventitious buds; and inoculating the cut adventitious buds on a rooting culture medium; and regularly culturing until white roots grow to obtain aseptic plantain germchits. According to the method provided by the invention, the disinfection efficiency of explants is up to 80% and the aseptic seedling rooting rate is 100%; the aseptic plantain germchits can be rapidly propagated; a plurality of problems that the propagation capability of plantain plants is low, the propagation period is long, resources are lacked and a lot of pesticides are remained are solved; the in-vitro rapid propagation method for the plantains has very important meanings in the aspects of protecting the natural resources of the wild plantains, protecting a good ecological environment and the like.
Description
Technical field
The invention belongs to the field of tissue culture of plant, be specifically related to a kind of method of Asiatic plantain Vitro Quick Reproduction.
Background technology
Plantago is in Plantaginaceae plant, and another name be again dish before Niu Tiancao, car, wheel dish, pig Auricled Hedyotis Herb etc., long in a humid environment, as pond, set under, the ground such as river bank, roadside.Asiatic plantain has edible and food therapy value, is rich in multivitamin and protein, containing enriching good soluble dietary fiber.
The seed of Asiatic plantain and herb all can be used as medicine, and toxic and side effect is less, are a kind of good medicinal plants, contain multiple medicinal component, as ursolic acid, plantagin, cupreol etc., have that inducing diuresis for removing edema, antidiarrheal, heat-clearing sterilization, improving eyesight reduce phlegm, an anti-inflammatory, the effect such as hypotensive; The oxygen radical that the β-Biao logaric acid containing (β-epiloganic acid), daucosterol and three kinds of compositions of galuteolin not only can be removed in body prevents aging, but also thering is antidepressant effect, the application of clinicing aspect has: treatment vaginitis, treatment acute mastitis, treatment postpartum retention of urine, treatment latent nephritis, treatment bedsore, treatment CAH, treatment exercise hemoglobinuria etc.; Also there is certain diuresis, can treat blood urine, flu and diarrhoea, treatment gynaecological disease (as leukorrhea), treatment intractable toothache.Also have at present and take Asiatic plantain and produce (as the ointment, syrup, health care buccal tablets, health drink etc.) of preparation as raw material.In addition, in Asiatic plantain, also contain the materials such as fatty acid, adenine such as the medicinal chemical compositions such as ursolic acid B-sitosterol, plantagin Aucubin, benzyl carbinol glycosides, Cobastab, positive hentriacontane and plantaglucide, Catalpol, choline, arachidic acid and linolenic acid.
Breeding for Asiatic plantain at present only has traditional planting technology.But because long-term over-exploitation causes, wild Asiatic plantain reduces gradually, seed carries virus, the shortcomings such as environmental condition deterioration causes the difference of medical value, breeding cycle is long, fine quality is degenerated and residue of pesticide amount exceeds standard, have all seriously affected the exploitation of Asiatic plantain.It is to solve the effective ways that current Asiatic plantain utilizes predicament that research Asiatic plantain is organized cultured in vitro quick propagating technology.
Summary of the invention
For solving the problems such as the seed existing in current Asiatic plantain utilization is taken poison, breeding cycle length, quality deterioration, medical value is uneven, residue of pesticide amount exceeds standard, the invention provides a kind of method of Asiatic plantain Vitro Quick Reproduction.
The method of Asiatic plantain Vitro Quick Reproduction provided by the invention, comprises the steps:
1) induction of callus: the Asiatic plantain blade through sterilization is cut to vein and leaf margin, be cut into small pieces, be inoculated on callus inducing medium, cellar culture is to forming callus and indefinite bud;
2) culture of rootage: select bud in step 1) and reach the indefinite bud of 2-3 centimetre, it is cut from callus, all the other callus and indefinite bud continue induction of callus, the indefinite bud cutting is inoculated on root media, cellar culture, to growing white root, obtains aseptic Asiatic plantain seedling.
Wherein, the concrete grammar of described sterilization is: pluck the Asiatic plantain blade without damage by disease and insect, flowing water rinses, and use alcohol-pickled 30s on superclean bench, then proceeds in 0.1% mercuric chloride solution and soak 8min, use aseptic water washing 3-5 time.
Wherein, the fritter of the preferred 0.8mm * 0.8mm of described fritter.
Wherein, described callus inducing medium is for take MS medium as minimal medium, and contained sucrose concentration is that 10～50g/L, methyl α-naphthyl acetate (NAA) concentration are the medium that 0.05～0.5mg/L, 6-benzyl aminopurine (6-BA) concentration are 1.0～4.0mg/L.
Preferably, described callus inducing medium is for take MS medium as minimal medium, contained sucrose concentration is that 30g/L, concentration of NAA are the medium that 0.1mg/L, 6-benzyl aminopurine concentration are 1.0mg/L, or take MS medium as minimal medium, contained sucrose concentration is that 30g/L, concentration of NAA are the medium that 0.1mg/L, 6-benzyl aminopurine concentration are 2.0mg/L.
Wherein, described root media is for take MS medium as minimal medium, and contained sucrose concentration is that 30g/L, concentration of NAA are the medium that 0.1mg/L, 6-benzyl aminopurine concentration are 2.0mg/L.
Wherein, the condition of culture of described cellar culture is pH value 6.0,25 ℃ of cultivation temperature, and periodicity of illumination 12 hours/day, intensity of illumination is 2000lx.
Wherein, the time of described induction of callus is 30 days.
Wherein, the time of described culture of rootage is 15 days.
The application of the method that the present invention also provides described Asiatic plantain Vitro Quick Reproduction in Asiatic plantain breeds.
Beneficial effect of the present invention is:
1) explant disinfection efficiency of the present invention is up to 80%, and aseptic seedling rooting rate reaches 100%, can the aseptic seedling of Fast-propagation Asiatic plantain.
2) the invention solves the problems such as the fertility of Asiatic plantain plant is low, breeding cycle length, scarcity of resources, residue of pesticide.
3) the present invention adopts Vitro Quick Reproduction technology to organize cultivation to Asiatic plantain first, not only can meet the need of market, but also can provide useful theoretical reference for factory's large-scale production.
4) the present invention is significant at the wild Asiatic plantain natural resources of protection and good aspects such as ecotope.
Embodiment
Following examples are used for illustrating the present invention, but are not used for limiting the scope of the invention.Without departing from the spirit and substance of the case in the present invention, the modification that the inventive method, step or condition are done or replacement, all belong to scope of the present invention.
If do not specialize, the conventional means that in embodiment, technological means used is well known to those skilled in the art.In embodiment, chemical reagent used is all commercially available.
Embodiment 1
Utilize the present invention to carry out Asiatic plantain Vitro Quick Reproduction.
The selection of explant and sterilization: from field, pluck the wild Asiatic plantain blade without damage by disease and insect, under flowing water, wash away dust, on superclean bench, first use alcohol-pickled 30s, proceed to again in 0.1% mercuric chloride solution and soak 8min, with aseptic water washing 3-5 time, finally under aseptic condition, cut vein and the edge of Herba Plantaginis extract sheet, be cut into the fritter of 0.8mm * 0.8mm.
1) induction of callus: 0.8mm * 0.8mm fritter that the Asiatic plantain blade through sterilization is cut into is inoculated on callus inducing medium, cellar culture.Described callus inducing medium is for take MS medium as minimal medium, and contained sucrose concentration is that 30g/L, concentration of NAA are the medium that 0.1mg/L, 6-benzyl aminopurine concentration are 1.0mg/L, and Medium's PH Value is 6.0.The condition of culture of described cellar culture is pH value 6.0,25 ℃ of cultivation temperature, and periodicity of illumination 12 hours/day, intensity of illumination is 2000lx.Cultivate 30 days, formed callus and a large amount of indefinite bud, wherein some indefinite bud reaches 2-3 centimetre.
2) culture of rootage: select bud in step 1) and reach the indefinite bud of 2-3 centimetre, it is cut from callus, and all the other callus and indefinite bud continue induction of callus, and the indefinite bud cutting is inoculated on root media, cellar culture, the same step 1) of condition of culture.Described root media is for take MS medium as minimal medium, and contained sucrose concentration is that 30g/L, concentration of NAA are the medium that 0.1mg/L, 6-benzyl aminopurine concentration are 2.0mg/L, and Medium's PH Value is 6.0.Cultivate 15 days, grown white root, obtain aseptic Asiatic plantain seedling.
Result shows, explant disinfection efficiency, up to 80%, can obtain aseptic indefinite bud fast, and rooting rate reaches 100%, a large amount of Asiatic plantain seedlings of the maternal plant merit that can be maintained.
Embodiment 2
Utilize the present invention to carry out Asiatic plantain Vitro Quick Reproduction.
Described callus inducing medium is for take MS medium as minimal medium, and contained sucrose concentration is that 30g/L, concentration of NAA are the medium that 0.1mg/L, 6-benzyl aminopurine concentration are 2.0mg/L, and Medium's PH Value is 6.0.Other conditions are with embodiment 1.
Result shows, explant disinfection efficiency, up to 80%, can obtain aseptic indefinite bud fast, and rooting rate reaches 100%, a large amount of Asiatic plantain seedlings of the maternal plant merit that can be maintained.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, do not departing under the prerequisite of the technology of the present invention principle; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (10)
1. a method for Asiatic plantain Vitro Quick Reproduction, comprises the steps:
1) induction of callus: the Asiatic plantain blade through sterilization is cut to vein and leaf margin, be cut into small pieces, be inoculated on callus inducing medium, cellar culture is to forming callus and indefinite bud;
2) culture of rootage: select bud in step 1) and reach the indefinite bud of 2-3 centimetre, it is cut from callus, all the other callus and indefinite bud continue induction of callus, the indefinite bud cutting is inoculated on root media, cellar culture, to growing white root, obtains aseptic Asiatic plantain seedling.
2. the method for Asiatic plantain Vitro Quick Reproduction as claimed in claim 1, it is characterized in that, the concrete grammar of described sterilization is: pluck the Asiatic plantain blade without damage by disease and insect, flowing water rinses, on superclean bench, use alcohol-pickled 30s, proceed to again in 0.1% mercuric chloride solution and soak 8min, with aseptic water washing 3-5 time.
3. the method for Asiatic plantain Vitro Quick Reproduction as claimed in claim 1, is characterized in that, the fritter of the preferred 0.8mm * 0.8mm of described fritter.
4. the method for Asiatic plantain Vitro Quick Reproduction as claimed in claim 1, it is characterized in that, described callus inducing medium is for take MS medium as minimal medium, and contained sucrose concentration is that 10～50g/L, concentration of NAA are the medium that 0.05～0.5mg/L, 6-benzyl aminopurine concentration are 1.0～4.0mg/L.
5. the method for Asiatic plantain Vitro Quick Reproduction as claimed in claim 4, it is characterized in that, described callus inducing medium is for take MS medium as minimal medium, contained sucrose concentration is that 30g/L, concentration of NAA are the medium that 0.1mg/L, 6-benzyl aminopurine concentration are 1.0mg/L, or take MS medium as minimal medium, contained sucrose concentration is that 30g/L, concentration of NAA are the medium that 0.1mg/L, 6-benzyl aminopurine concentration are 2.0mg/L.
6. the method for Asiatic plantain Vitro Quick Reproduction as claimed in claim 1, it is characterized in that, described root media is for take MS medium as minimal medium, and contained sucrose concentration is that 30g/L, concentration of NAA are the medium that 0.1mg/L, 6-benzyl aminopurine concentration are 2.0mg/L.
7. the method for Asiatic plantain Vitro Quick Reproduction as claimed in claim 1, is characterized in that, the condition of culture of described cellar culture is pH value 6.0,25 ℃ of cultivation temperature, and periodicity of illumination 12 hours/day, intensity of illumination is 2000lx.
8. the method for Asiatic plantain Vitro Quick Reproduction as claimed in claim 1, is characterized in that, the time of described induction of callus is 28-32 days.
9. the method for Asiatic plantain Vitro Quick Reproduction as claimed in claim 1, is characterized in that, the time of described culture of rootage is 14-16 days.
10. the application of the method for the Asiatic plantain Vitro Quick Reproduction described in claim 1-9 any one in Asiatic plantain breeds.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108260526A (en) * | 2017-12-05 | 2018-07-10 | 长沙学院 | A kind of method for tissue culture of caducous Mazus japonicus |
| CN115474551A (en) * | 2022-10-18 | 2022-12-16 | 中国长江三峡集团有限公司 | Tissue culture and rapid propagation method of national second-level protection plant of Tokyo plantago |
| CN115868408A (en) * | 2022-10-18 | 2023-03-31 | 中国长江三峡集团有限公司 | Method for efficient breeding of endangered plant Plantago Tomentosa |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1685811A (en) * | 2005-06-03 | 2005-10-26 | 中国科学院植物研究所 | In vitro breeding method of plantage |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1685811A (en) * | 2005-06-03 | 2005-10-26 | 中国科学院植物研究所 | In vitro breeding method of plantage |
Non-Patent Citations (4)
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| EMILIA ANDRZEJEWSKA-GOLEC ETAL: "MICROPROPAGATION OF PLANTAGO CAMTSCHATICA LINK", 《ACTA SOCIETATIS BOTANICORUM POLONIAE》 * |
| 曾建军等: "车前不定芽直接诱导及再生体系的建立", 《安徽农业科学》 * |
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| 涂艺声: "车前的离体培养和植株再生", 《江西科学》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108260526A (en) * | 2017-12-05 | 2018-07-10 | 长沙学院 | A kind of method for tissue culture of caducous Mazus japonicus |
| CN108260526B (en) * | 2017-12-05 | 2021-01-12 | 长沙学院 | Tissue culture method of early-falling marsdenia tenacissima |
| CN115474551A (en) * | 2022-10-18 | 2022-12-16 | 中国长江三峡集团有限公司 | Tissue culture and rapid propagation method of national second-level protection plant of Tokyo plantago |
| CN115868408A (en) * | 2022-10-18 | 2023-03-31 | 中国长江三峡集团有限公司 | Method for efficient breeding of endangered plant Plantago Tomentosa |
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