CN103070073A - Method for fast propagation of test tube arrowhead by tissue culture - Google Patents
Method for fast propagation of test tube arrowhead by tissue culture Download PDFInfo
- Publication number
- CN103070073A CN103070073A CN2013100242769A CN201310024276A CN103070073A CN 103070073 A CN103070073 A CN 103070073A CN 2013100242769 A CN2013100242769 A CN 2013100242769A CN 201310024276 A CN201310024276 A CN 201310024276A CN 103070073 A CN103070073 A CN 103070073A
- Authority
- CN
- China
- Prior art keywords
- arrowhead
- test tube
- mass fraction
- medium
- propagation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a method for fast propagation of test tube arrowhead by tissue culture. The test tube arrowhead is formed by sterilizing treatment of an explant, stem tip differentiation, subculture multiplication and induction. According to the method disclosed by the invention, the propagation of the test tube arrowhead is carried by tissue culture, and thus high propagation coefficient and short propagation period are realized; in addition, the whole propagation process is carried out under manual control conditions without being influenced by an external environment; the test tube arrowhead can be annually produced; due to small size and light weight, the test tube arrowhead can solve the problems existing in the remote transportation and storage of arrowhead test tube seedlings; the test tube arrowhead has no disease, so that the problem that a conventional arrowhead seed easily carries diseases is solved; the induction of the test tube arrowhead provides an operable simple system for research work of the carmus expansion mechanism; as an in-vitro dormancy organ, the in-vitro indoor preservation research on arrowhead germ plasm resources can be realized; and live exchange of international germ plasm resources is benefited. The invention provides a quick, convenient and effective propagation method for the development of the arrowhead industry.
Description
Technical field
The invention belongs to the agricultural breeding field, refer to particularly a kind of method of test tube arrowhead tissue-culturing quick-propagation.
Background technology
Arrowhead is perennial aquatic herbaceous plant, and is edible with its bulb, and China is one of arrowhead original producton location, the Yangtze river basin and on the south the each province generally cultivate, Taihu Lake bank and the Delta of the Pearl River are the main producing region, cultivate on a small quantity in the north.Arrowhead mainly does to plant with arrowhead bulb or bulb terminal bud and carries out vegetative propagation, and reproduction coefficient is low, only breeds a generation in 1 year, and sowing quantity is large, has limited the promotion rate of the new quality product kind of arrowhead.
Therefore, the production of carrying out the test tube arrowhead by tissue-culturing quick-propagation is one of effective way that addresses the above problem.Present stage, the Study on tissue culture work of arrowhead is mainly undertaken by domestic researcher, and less to its research abroad.Guizhou Normal University etc. have carried out some research work, but have to arrowhead test-tube plantlet and stolon.Arrowhead test-tube plantlet and stolon are unfavorable for long distance transportation and storage.
Summary of the invention
The objective of the invention is sick for the kind spherical zone of vegetative arrowhead and kind sexual involution problem, a kind of method of test tube arrowhead tissue-culturing quick-propagation is provided, improve the quality of arrowhead.
The method of test tube arrowhead tissue-culturing quick-propagation of the present invention may further comprise the steps:
1) disinfecting of explant: the arrowhead bulb of choosing anosis worm harm, after cleaning up, cutting the bulb terminal bud is explant, flowing water flushing 20～35min, peel off 1～2 layer of scale disinfection of outermost after being filtered dry, being that 72% streptomycin sulphate is processed 10～30min with mass fraction first, is 70% ethanol surface sterilization, 20～40s with mass fraction again, is 0.1%HgCl with mass fraction at last
2Solution-treated 2～5min;
2) stem apex differentiation: aseptic water washing 4～5 times of the arrowhead after disinfecting, peel off 2～3 layers of scale, cutting stem apex is inoculated on the startup medium, described startup medium is the 1/2MS medium, add 6-benzyl aminopurine (6-BA) 1.0～2.0mg/L, methyl α-naphthyl acetate (NAA) 0.1～0.5mg/L, mass fraction is 3.0% sucrose and mass fraction is 0～0.5% agar, the pH value is 5.8～6.0,25 ℃～28 ℃ of cultivation temperature, intensity of illumination 1000～1500 lx, every day light application time 10h, until grow up to Regenerated plantlet;
3) shoot proliferation: the Regenerated plantlet that stem apex is induced is transferred to and carries out shoot proliferation in the proliferated culture medium and cultivate, described proliferated culture medium is the MS medium, add 6-benzyl aminopurine (6-BA) 1.0～3.0 mg/L, methyl α-naphthyl acetate (NAA) 0.1～0.3mg/L, mass fraction is 3.0% sucrose and mass fraction is 0～0.5% agar, the pH value is 5.8～6.0,25 ℃～28 ℃ of cultivation temperature, intensity of illumination 1000～1500 lx, every day light application time 10h, until sprout to form the arrowhead test-tube plantlet of plant division;
4) induce the test tube arrowhead: the arrowhead test-tube plantlet is transferred on the inducing culture, described inducing culture is the MS medium, add 6-benzyl aminopurine (6-BA) 0～3.0mg/L, methyl α-naphthyl acetate (NAA) 0～0.3mg/L, active carbon (AC) 0.5～1.0g/L, mass fraction are 3.0%～8.0% sucrose and mass fraction is 0～0.5% agar, the pH value is 5.8～6.0,14 ℃～22 ℃ of cultivation temperature, every day light application time 0～12h condition under induce and form the test tube arrowhead.
The present invention is in the disinfecting of explant, and having prolonged mass fraction is the processing time of 72% streptomycin sulphate, the corresponding processing time that shortens 0.1% mercuric chloride.Because mercuric chloride (HgCl
2) toxicity very large, when explant is carried out disinfection, also explant is caused very large injury, heavy then cause material dead, so after the sterilization treatment step is optimized, arrowhead stem apex survival rate can have been improved 5～10 percentage points.
The present invention is at stem apex in differential period, and the minimal medium of selecting the differentiation of arrowhead stem apex to start in the medium is the 1/2MS medium, and the macroelement of MS is reduced by half.Like this, more be conducive to the differentiation of root owing to having reduced the concentration of mineral salt, thereby arrowhead stem apex differentiation rate has been improved 2～3 percentage points, also reduced the cost of medium simultaneously.
The present invention is at shoot proliferation in the stage, because 6-BA concentration height easily causes the differentiation of tissue cultured test-tube seedling too much, seedling is little, and growth is slow, and lopsided seedling is arranged, and the accumulation of 6-BA easily causes the vitrifying of tissue cultured test-tube seedling, cause tissue cultured test-tube seedling lethality high, and proterties is difficult to stablize.Therefore, by optimizing the concentration of 6-BA in the proliferated culture medium, in reducing proliferated culture medium, can improve again the survival rate of tissue cultured test-tube seedling in the concentration of 6-BA.
The present invention has added active carbon AC inducing in the test tube arrowhead stage in inducing culture.The interpolation of AC provides not only for the test tube arrowhead more to be suitable for the dark situation of inducing, the mortifier in the adsorbable cultured in vitro also, simultaneously AC also produces certain suction-operated to the 6-BA of high concentration and NAA etc., is conducive to induce test tube arrowhead more, larger and that regularity is high.
The present invention cultivates by tissue and carries out the breeding of test tube arrowhead, has the following advantages:
The first, reproduction coefficient is high, and the breeding cycle is short, and whole reproductive process all carries out under manually operated condition, is not affected by the external environment, and can carry out the anniversary and produce, and can breed for 6～8 generations in 1 year; And conventional the production with planting of arrowhead bred a generation in 1 year only.The present invention has greatly improved the reproduction speed of arrowhead, is conducive to applying of excellent new arrowhead kind.
The second, the test tube arrowhead is little, lightweight because of its volume, both has been convenient to long-distance transport, reduced again with kind the proportion of goods damageds saved freight, solved the problem that the arrowhead test-tube plantlet is unfavorable for long-distance transport and storage.The single heavy 25～40g of conventional arrowhead, every mu of sowing quantity of traditional mode of production mode is 100kg, and single test tube arrowhead only weighs 0.1～0.3g, every mu of sowing quantity 0.4～1.2kg can minimize transport costs more than 80%, reduces with kind of the proportion of goods damageds more than 20%.
The 3rd, the test tube arrowhead has solved conventional kind of the easy problem in spite of illness of using of arrowhead not in spite of illness.By the fast numerous test tube arrowhead that obtains of Shoot-tip Culture not in spite of illness, the field growing resistance against diseases is strong, shows as plant tall and big, and the leaf look dark green, and the stem stalk is sturdy, and growth potential is strong.
The 4th, inducing of test tube arrowhead will provide an exercisable single system for the work of expansion of corms research on mechanism.
The 5th, as stripped dormancy organ, can also carry out the stripped indoor preservation research of arrowhead germ plasm resource, and be conducive to the live body exchange of international germ plasm resource.
In sum, the present invention provides a kind of quick, convenient, effective propagation method for the development of arrowhead industry.
Embodiment
The present invention is described in further detail below in conjunction with specific embodiment.
Embodiment 1
The terminal bud of purple circle arrowhead (being commonly called as " crow of peeling old man ") is as explant take Jiangsu Local kind Baoying County, through the disinfecting of explant, stem apex differentiation, shoot proliferation, induce and form the test tube arrowhead, comprises the steps:
1) disinfecting of explant: the arrowhead bulb of choosing anosis worm harm, after cleaning up, cutting the bulb terminal bud is explant, flowing water washes approximately 30min, be filtered dry and peel off 2 layers of scale disinfection of outermost on the rearmounted superclean bench, being that 72% streptomycin sulphate is processed 30min with mass fraction first namely, is 70% ethanol surface sterilization 30s with mass fraction again, is 0.1%HgCl with mass fraction at last
2Solution-treated 3min;
2) stem apex differentiation: aseptic water washing 5 times of the arrowhead after disinfecting, peel off 3 layers of scale with scalpel, stem apex is peeled off to only remaining 2 leaf primordium, cut stem apex 0.5cm and be inoculated on the startup medium.Used startup medium is the 1/2MS medium, add 6-benzyl aminopurine (6-BA) 1.0mg/L, methyl α-naphthyl acetate (NAA) 0.2mg/L, mass fraction is 3.0% sucrose, the pH value is 6.0, it is 28 ℃ in temperature, intensity of illumination is under 1000 lx, illumination every day 10h, 30d can grow up to the Regenerated plantlet with 4 leaves;
3) shoot proliferation: the Regenerated plantlet that stem apex is induced is transferred to and carries out shoot proliferation in the proliferated culture medium and cultivate, used proliferated culture medium is the MS medium, add 6-benzyl aminopurine (6-BA) 2.0 mg/L, methyl α-naphthyl acetate (NAA) 0.1mg/L, mass fraction is 3.0% sucrose, the pH value is 6.0, it is 28 ℃ in temperature, intensity of illumination is under 1200 lx, illumination every day 10h, 40d sprouts the arrowhead test-tube plantlet that forms plant division;
4) induce the test tube arrowhead: the arrowhead test-tube plantlet is transferred on the inducing culture, used inducing culture is the MS medium, add 6-benzyl aminopurine (6-BA) 1.5mg/L, methyl α-naphthyl acetate (NAA) 0.1mg/L, active carbon (AC) 1.0g/L, mass fraction are 6.0% sucrose, the pH value is 6.0,22 ℃ of temperature, under the full dark condition, 60d induces later on and forms the test tube arrowhead.
Embodiment 2
The terminal bud of purple circle arrowhead (being commonly called as " crow of peeling old man ") is as explant take Jiangsu Local kind Baoying County, through the disinfecting of explant, stem apex differentiation, shoot proliferation, induce and form the test tube arrowhead, comprises the steps:
1) disinfecting of explant: the arrowhead bulb of choosing anosis worm harm, after cleaning up, cutting the bulb terminal bud is explant, flowing water washes approximately 25min, be filtered dry and peel off 2 layers of scale disinfection of outermost on the rearmounted superclean bench, being that 72% streptomycin sulphate is processed 20min with mass fraction first namely, is 70% ethanol surface sterilization 40s with mass fraction again, is 0.1%HgCl with mass fraction at last
2Solution-treated 5min;
2) stem apex differentiation: aseptic water washing 5 times of the arrowhead after disinfecting, peel off 3 layers of scale with scalpel, stem apex is peeled off to only remaining 2 leaf primordium, cut stem apex 0.5cm and be inoculated on the startup medium.Used startup medium is the 1/2MS medium, add 6-benzyl aminopurine (6-BA) 1.5mg/L, methyl α-naphthyl acetate (NAA) 0.4mg/L, mass fraction is 3.0% sucrose and mass fraction is 0.5% agar, the pH value is 5.8, be 25 ℃ ℃ in temperature, intensity of illumination is under 1000 lx, illumination every day 10h, 32d can grow up to the Regenerated plantlet with 7 leaves;
3) shoot proliferation: the Regenerated plantlet that stem apex is induced is transferred to and carries out shoot proliferation in the proliferated culture medium and cultivate, used proliferated culture medium is the MS medium, add 6-benzyl aminopurine (6-BA) 1.0 mg/L, methyl α-naphthyl acetate (NAA) 0.2mg/L, mass fraction is 3.0% sucrose and mass fraction is 0.2% agar, the pH value is 5.8, it is 25 ℃ in temperature, intensity of illumination is under 1500 lx, illumination every day 10h, 35d sprouts the arrowhead test-tube plantlet that forms plant division;
4) induce the test tube arrowhead: the arrowhead test-tube plantlet is transferred on the inducing culture, used inducing culture is the MS medium, add 6-benzyl aminopurine (6-BA) 0.5mg/L, methyl α-naphthyl acetate (NAA) 0.1mg/L, active carbon (AC) 1.0g/L, mass fraction are 7.0% sucrose and mass fraction is 0.3% agar, the pH value is 5.8,20 ℃ of temperature, every day light application time 4h condition under, 55d induces later on and forms the test tube arrowhead.
Embodiment 3
The terminal bud of purple circle arrowhead (being commonly called as " crow of peeling old man ") is as explant take Jiangsu Local kind Baoying County, through the disinfecting of explant, stem apex differentiation, shoot proliferation, induce and form the test tube arrowhead, comprises the steps:
1) disinfecting of explant: the arrowhead bulb of choosing anosis worm harm, after cleaning up, cutting the bulb terminal bud is explant, flowing water washes approximately 35min, be filtered dry and peel off 1 layer of scale disinfection of outermost on the rearmounted superclean bench, being that 72% streptomycin sulphate is processed 10min with mass fraction first namely, is 70% ethanol surface sterilization 30s with mass fraction again, is 0.1%HgCl with mass fraction at last
2Solution-treated 2min;
2) stem apex differentiation: aseptic water washing 5 times of the arrowhead after disinfecting, peel off 3 layers of scale with scalpel, stem apex is peeled off to only remaining 1 leaf primordium, cut stem apex 0.5cm and be inoculated on the startup medium.Used startup medium is the 1/2MS medium, add 6-benzyl aminopurine (6-BA) 2.0mg/L, methyl α-naphthyl acetate (NAA) 0.4mg/L, mass fraction is 3.0% sucrose and mass fraction is 0.2% agar, the pH value is 6.0, it is 28 ℃ in temperature, intensity of illumination is under 1200 lx, illumination every day 10h, 35d can grow up to the Regenerated plantlet with 4 leaves;
3) shoot proliferation: the Regenerated plantlet that stem apex is induced is transferred to and carries out shoot proliferation in the proliferated culture medium and cultivate, used proliferated culture medium is the MS medium, add 6-benzyl aminopurine (6-BA) 3.0 mg/L, methyl α-naphthyl acetate (NAA) 0.2mg/L, mass fraction is 3.0% sucrose and mass fraction is 0.4% agar, the pH value is 5.8～6.0, be 25 ℃ ℃ in temperature, intensity of illumination is under 1500 lx, illumination every day 10h, 40d sprouts the arrowhead test-tube plantlet that forms plant division;
4) induce the test tube arrowhead: the arrowhead test-tube plantlet is transferred on the inducing culture, used inducing culture is the MS medium, add 6-benzyl aminopurine (6-BA) 3mg/L, active carbon (AC) 0.5g/L, mass fraction is 8.0% sucrose and mass fraction is 0.3% agar, the pH value is 6.0,20 ℃ of temperature, every day light application time 8h condition under, 55d induces later on and forms the test tube arrowhead.
Embodiment 4
The terminal bud of purple circle arrowhead (being commonly called as " crow of peeling old man ") is as explant take Jiangsu Local kind Baoying County, through the disinfecting of explant, stem apex differentiation, shoot proliferation, induce and form the test tube arrowhead, comprises the steps:
1) disinfecting of explant: the arrowhead bulb of choosing anosis worm harm, after cleaning up, cutting the bulb terminal bud is explant, flowing water washes approximately 30min, be filtered dry and peel off 1 layer of scale disinfection of outermost on the rearmounted superclean bench, being that 72% streptomycin sulphate is processed 15min with mass fraction first namely, is 70% ethanol surface sterilization 25s with mass fraction again, is 0.1%HgCl with mass fraction at last
2Solution-treated 4min;
2) stem apex differentiation: aseptic water washing 4 times of the arrowhead after disinfecting, peel off 3 layers of scale with scalpel, stem apex is peeled off to only remaining 2 leaf primordium, cut stem apex 0.5cm and be inoculated on the startup medium.Used startup medium is the 1/2MS medium, add 6-benzyl aminopurine (6-BA) 1.7mg/L, methyl α-naphthyl acetate (NAA) 0.3mg/L, mass fraction is 3.0% sucrose, the pH value is 6.0, it is 25 ℃ in temperature, intensity of illumination is under 1300 lx, illumination every day 10h, 35d can grow up to the Regenerated plantlet with 6 leaves;
3) shoot proliferation: the Regenerated plantlet that stem apex is induced is transferred to and carries out shoot proliferation in the proliferated culture medium and cultivate, used proliferated culture medium is the MS medium, add 6-benzyl aminopurine (6-BA) 2.2 mg/L, methyl α-naphthyl acetate (NAA) 0.1mg/L, mass fraction is 3.0% sucrose and mass fraction is 0.1% agar, the pH value is 6.0, it is 25 ℃ in temperature, intensity of illumination is under 1400 lx, illumination every day 10h, 40d sprouts the arrowhead test-tube plantlet that forms plant division;
4) induce the test tube arrowhead: the arrowhead test-tube plantlet is transferred on the inducing culture, used inducing culture is the MS medium, add methyl α-naphthyl acetate (NAA) 0.3mg/L, active carbon (AC) 0.8g/L, mass fraction is 5.0% sucrose, the pH value is 6.0,14 ℃ of temperature, every day light application time 12h condition under, 60d induces later on and forms the test tube arrowhead.
Claims (1)
1. the method for a test tube arrowhead tissue-culturing quick-propagation is characterized in that, may further comprise the steps:
1) disinfecting of explant: the arrowhead bulb of choosing anosis worm harm, after cleaning up, cutting the bulb terminal bud is explant, flowing water flushing 20～35min, peel off 1～2 layer of scale disinfection of outermost after being filtered dry, being that 72% streptomycin sulphate is processed 10～30min with mass fraction first, is 70% ethanol surface sterilization, 20～40s with mass fraction again, is 0.1%HgCl with mass fraction at last
2Solution-treated 2～5min;
2) stem apex differentiation: aseptic water washing 4～5 times of the arrowhead after disinfecting, peel off 2～3 layers of scale, cutting stem apex is inoculated on the startup medium, described startup medium is the 1/2MS medium, add 6-benzyl aminopurine 1.0～2.0mg/L, methyl α-naphthyl acetate 0.1～0.5mg/L, mass fraction is 3.0% sucrose and mass fraction is 0～0.5% agar, the pH value is 5.8～6.0,25 ℃～28 ℃ of cultivation temperature, intensity of illumination 1000～1500 lx, every day light application time 10h, until grow up to Regenerated plantlet;
3) shoot proliferation: the Regenerated plantlet that stem apex is induced is transferred to and carries out shoot proliferation in the proliferated culture medium and cultivate, described proliferated culture medium is the MS medium, add 6-benzyl aminopurine 1.0～3.0 mg/L, methyl α-naphthyl acetate 0.1～0.3mg/L, mass fraction is 3.0% sucrose and mass fraction is 0～0.5% agar, the pH value is 5.8～6.0,25 ℃～28 ℃ of cultivation temperature, intensity of illumination 1000～1500 lx, every day light application time 10h, until sprout to form the arrowhead test-tube plantlet of plant division;
4) induce the test tube arrowhead: the arrowhead test-tube plantlet is transferred on the inducing culture, described inducing culture is the MS medium, add 6-benzyl aminopurine 0～3.0mg/L, methyl α-naphthyl acetate 0～0.3mg/L, active carbon 0.5～1.0g/L, mass fraction are 3.0%～8.0% sucrose and mass fraction is 0～0.5% agar, the pH value is 5.8～6.0,14 ℃～22 ℃ of cultivation temperature, every day light application time 0～12h condition under induce and form the test tube arrowhead.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2013100242769A CN103070073A (en) | 2013-01-22 | 2013-01-22 | Method for fast propagation of test tube arrowhead by tissue culture |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2013100242769A CN103070073A (en) | 2013-01-22 | 2013-01-22 | Method for fast propagation of test tube arrowhead by tissue culture |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103070073A true CN103070073A (en) | 2013-05-01 |
Family
ID=48146941
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2013100242769A Pending CN103070073A (en) | 2013-01-22 | 2013-01-22 | Method for fast propagation of test tube arrowhead by tissue culture |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103070073A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104278007A (en) * | 2014-10-27 | 2015-01-14 | 扬州大学 | Preparation method of arrowhead protoplasts |
| CN104542277A (en) * | 2014-12-19 | 2015-04-29 | 广西壮族自治区农业科学院生物技术研究所 | Rapid propagation method for sagittaria sagittifolia tissue culture seedling and culture medium |
| CN108703068A (en) * | 2018-04-04 | 2018-10-26 | 广西壮族自治区农业科学院生物技术研究所 | Remove method, cultural method and the application of endophyte in arrowhead incubation |
| CN108812305A (en) * | 2018-04-04 | 2018-11-16 | 广西壮族自治区农业科学院生物技术研究所 | Obtain the method and its application of sterile arrowhead explant |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101554137A (en) * | 2009-04-09 | 2009-10-14 | 白永兴 | Tissue culture method of arrowhead |
| WO2011079212A2 (en) * | 2009-12-24 | 2011-06-30 | LifeSpan Extension, LLC | Methods and compositions for identifying, producing and using plant-derived products modulating cell function and aging |
| CN102369825A (en) * | 2010-08-10 | 2012-03-14 | 徐一华 | High-yield arrowhead culture method |
-
2013
- 2013-01-22 CN CN2013100242769A patent/CN103070073A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101554137A (en) * | 2009-04-09 | 2009-10-14 | 白永兴 | Tissue culture method of arrowhead |
| WO2011079212A2 (en) * | 2009-12-24 | 2011-06-30 | LifeSpan Extension, LLC | Methods and compositions for identifying, producing and using plant-derived products modulating cell function and aging |
| CN102369825A (en) * | 2010-08-10 | 2012-03-14 | 徐一华 | High-yield arrowhead culture method |
Non-Patent Citations (1)
| Title |
|---|
| 朱红莲: "试管慈姑诱导的初步研究", 《华中农业大学硕士学位论文》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104278007A (en) * | 2014-10-27 | 2015-01-14 | 扬州大学 | Preparation method of arrowhead protoplasts |
| CN104278007B (en) * | 2014-10-27 | 2017-12-29 | 扬州大学 | A kind of preparation method of arrowhead protoplast |
| CN104542277A (en) * | 2014-12-19 | 2015-04-29 | 广西壮族自治区农业科学院生物技术研究所 | Rapid propagation method for sagittaria sagittifolia tissue culture seedling and culture medium |
| CN108703068A (en) * | 2018-04-04 | 2018-10-26 | 广西壮族自治区农业科学院生物技术研究所 | Remove method, cultural method and the application of endophyte in arrowhead incubation |
| CN108812305A (en) * | 2018-04-04 | 2018-11-16 | 广西壮族自治区农业科学院生物技术研究所 | Obtain the method and its application of sterile arrowhead explant |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102301951B (en) | Method for rapidly propagating roots of subprostrate sophora by tissue culture | |
| CN103250645B (en) | Rapid propagation and transplantation method of blumea balsamifera | |
| CN102550413A (en) | Rapid propagation method for tube seedlings of high-grade polygonatum cyrtonema hua | |
| CN108338071A (en) | A kind of asexual multiplication seedling method of hybridization paper mulberry | |
| CN101720670B (en) | Rapid breeding method for pinellia tuber tissue culture | |
| CN108157180B (en) | Open type factory rapid propagation method for potato virus-free seedlings | |
| CN101796924B (en) | Method for improving annular stalk growing rate of oriental lily test tube bulbs | |
| CN103168692B (en) | Salix saposhnikovii tissue culture method | |
| CN103070073A (en) | Method for fast propagation of test tube arrowhead by tissue culture | |
| CN100394845C (en) | In-bottle production method of detoxified small seed ball of east lily | |
| CN102499082A (en) | Test tube breeding method of lilium oriental hybrid seed ball | |
| CN103548695B (en) | A kind of meadowrueleaf corydalis root quick breeding method for tissue culture | |
| CN102860257A (en) | Houttuynia cordata aerial stem tissue culture rapid propagation method | |
| CN105028206A (en) | Tissue culture propagation method for Hainan dendrobe | |
| CN107873517A (en) | A kind of method by Chinese herbaceous peony cotyledon evoked callus regenerated adventitious bud | |
| CN103461126A (en) | Rapid propagation method for palm trees | |
| CN102138527B (en) | Method for culturing tissue culture seedlings of glabrous greenbrier rhizome | |
| CN103229721A (en) | Tissue culture propagation method of Gynura formosana | |
| CN104012406A (en) | Regeneration in-vitro method for sweet cherry variety wanhongzhu | |
| CN1918972B (en) | Angelica tissue culture breeding method | |
| CN107568065A (en) | A kind of hair Chinese in-vitro culture methods of volume 12 | |
| CN104285816A (en) | Rapid propagation method for xanthoceras sorbifolia bunge tissue during culturing | |
| CN105230488B (en) | A kind of Cymbidium lancifolium leaf tissue culture method for quickly breeding | |
| CN104221851B (en) | A kind of great Ye ant tower isolated culture and rapid propagation method | |
| CN105104197A (en) | Method for culturing and breeding dendrobium guangxiense tissue |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20130501 |