CN1204453A - Method for quick reproducing 'Shankui' sprout - Google Patents
Method for quick reproducing 'Shankui' sprout Download PDFInfo
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- CN1204453A CN1204453A CN 98103484 CN98103484A CN1204453A CN 1204453 A CN1204453 A CN 1204453A CN 98103484 CN98103484 CN 98103484 CN 98103484 A CN98103484 A CN 98103484A CN 1204453 A CN1204453 A CN 1204453A
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Abstract
A method for quickly reproducing the seed breedings of Shangkui (a herbaceous plant) includes such technological steps as taking the terminal buds or lateral buds of tuber by cutting in aseptic condition, selecting culture medium, proportioning hormone, changing culture condition, inducing seedlings in test tubes, quick reproducing the seedlings in test tubes, making the seedlings stronger, growing root and transplanting, and features high reproducing coefficient up to 4.0-5.0, high culture speed and low cost.
Description
The invention belongs to the quick propagating technology field of plant seedling, it is to be exclusively used in that horseradish (scurvy grass) tissue cultivating seedling is bred fast and the technology of test-tube plantlet field cultivation.
Horseradish (Wasabi) formal name used at school is Eutrema wasabi Maxim; Wasabi japonicamatum, have another name called scurvy grass, be Cruciferae perennial herb half shade plant, be the high plant of a kind of economic worth, its root-like stock, blade, petiole have abundant nutrition, and be not only edible, and has a very high medical value, its root-like stock is worn into and stuck with paste is the indispensable flavouring of seafood such as the edible raw fish of compatriots such as Japan and Southeast Asia, has strong sterilization and aid digestion function, and the good effect of preventing and curing diseases is arranged.Price is very expensive in the international market, and per kilogram root-like stock wholesale price is 40 dollars (nineteen ninety-fives) on the wholesale market, Tokyo.And supply falls short of demand, and the development horseradish produces foreign exchange earning can produce huge economic benefit.
Horseradish grows environmental condition is required very strictness, the shady and cool moistening condition of happiness, and non-refractory and low temperature were avoided intense light irradiation.Cultivation is difficulty quite, horseradish sapling multiplication difficulty in addition, and reproduction speed is slow, so its price is high.Thereby China 1994 beginning introducing and planting horseradish is badly in need of a large amount of seedlings, and producing at present mainly is to utilize from external import seed production, and price is very high, and average every strain 15-20 unit does not meet the requirement of production.The horseradish sapling multiplication is mainly undertaken by the method for nourishing and generating, reproduction coefficient is lower, reproduction speed is slower, one strain horseradish breeds in 1 year by offshoot and can only breed about 20 strains at most, is difficult to satisfy the requirement of large-scale production, and utilizing tissue culture method is the effective way that solves the sapling multiplication problem, method by tissue culture not only can obtain a large amount of nursery stocks at short notice, and after the aseptic seedling transplanting, growth is difficult for susceptible rapidly.According to the result of Agricultural University Of Nanjing's light disk retrieval in conjunction with manual information retrieval, domestic still nobody studies the tissue culture of horseradish and reports at present.Have only Japanology and report more abroad.But utilize Shoot Tip Culture, all about 3.0, used medium is MS+6-BA0.1mg/l+ sucrose 2% (agricultural and gardening, 1986,61 (8)) to its reproduction coefficient; MS+6-BA0.2mg/l+ sucrose 3% (garden is learned assorted, 62 other (2) 1993).Though above result of study has certain reference value, but produce for commercialization, it is lower that its reproduction coefficient still shows, it is still higher to produce the seedling cost with their method, can not obtain enough seedlings at short notice, therefore be badly in need of that a kind of reproduction coefficient is higher, speed faster propagation method carry out seedling production.
Purpose of the present invention just provides a kind of method for quickly breeding of horseradish seedling, makes the propagation of horseradish tissue cultivating seedling rapid, and its reproduction coefficient can reach 4.0-5.0, the test-tube plantlet that enrichment culture forms is directly used in production, saves labor, and economizes ground, speed is fast, reduces production costs.
The quick propagation method of horseradish tissue cultivating seedling provided by the present invention, the concrete operations step is as follows:
1) aseptic seedling is induced: cut the lateral bud on horseradish terminal bud or the root-like stock under the aseptic condition, be inoculated in MS (or B
5)+6-BA0.1-1mg/l+NAA0.05mg/l+GA
30.05-0.2mg/l+ carbohydrate 2-3% differentiates young shoot after 3～5 weeks of inoculation;
2) test-tube plantlet propagation: change test-tube plantlet over to MS (or B
5)+6-BA0.1-2mg/l+KT0.1-2mg/l+NAA0.01-0.5mg/l or IBA0.01-0.5mg/l or IAA0.01-0.5mg/l+GA
30.01-0.5mg/l on the proliferated culture medium, add carbohydrate 3-5%, inoculation back young shoot blade base axillalry bud is sprouted gradually, the switching back can form bud clump, shoot proliferation in 25-35 days;
3) culture of rootage: the no offspring of enrichment culture is inoculated in MS (or B after cutting
5On the root media of .2mg/l of)+NAA0.01-) or IBA0.01-0.1mg/l or IAA0.01-0.2mg/l+ carbohydrate 2-3%, taking root through 15-25 days seedling base portions becomes whole plant;
4) agar content is 0.6-0.8% in above-mentioned each medium, pH5.8-6.2.
The method for quickly breeding of above-mentioned horseradish seedling, wherein each stage condition of culture is that cultivation temperature is 18-22 ℃, intensity of illumination 1500-2000Lux, irradiation 14-16 hour/day.Used carbohydrate is meant sucrose, glucose, fructose etc., is best with sucrose.
In the shoot proliferation process, increase test-tube plantlet strong seedling culture process, change test-tube plantlet over to strong seedling culture base: MS or B
5+ 6-BA0.1-1mg/l+KT0.1-1mg/l+NAA0.05mg/l+GA
30.1-0.5mg/l+ carbohydrate 3-4% promotes the blade petiole to stretch, and changes above-mentioned proliferated culture medium again over to, promotes its differentiation.
The present invention compared with prior art has following advantage:
1. compare with the technology of direct offshoot breeding horseradish seedling and existing tissue culture technique breeding horseradish seedling, method is simple, be not subject to seasonal restrictions, combine by multiple hormone, enrichment culture and strong seedling culture alternate, and methods such as osmotic adjustment and change condition of culture cultivate, and its reproduction speed is faster, and reproductive efficiency is higher.Utilize the present invention, a strain horseradish test-tube plantlet can be bred the 4-5 strain in 30 days, can breed continuously 9-10 generation in 1 year, can produce test-tube plantlet 4
9-10Strain, and strain breeding 20 strains at most in the conventional offshoot 1 year utilize breeding 3 at most in the existing tissue culture technique 1 year
9-10Strain, reproduction speed increases significantly, and more meets the requirement that commercialization is produced.
2. compare with test-tube plantlet domestication and the cultivation method reported, the test-tube plantlet survival rate is higher, can reach more than 85%, and only need simple facility, and cost is lower, and method is simple, and is practical reliable.
3. breed by Shoot Tip Culture, the hereditary capacity of horseradish seedling is stable, can not morph.Be fit to commercialization production.
4. propagation method provided by the present invention and culture technique are applicable to various types of horseradish kinds.
Embodiment 1:
1. test-tube plantlet is induced: cut " No. 3, Shimane " horseradish root-like stock lateral bud, be cut into the 0.5cm size under the aseptic condition, be inoculated in inducing culture, the inducing culture based formulas is MS+6-BA0.2mg/l+NAA0.1mg/l+GA
30.2mg/l+ sucrose 3%+ agar powder 0.65%, pH6.0, condition of culture be, 20 ℃ of temperature, and illumination 1500Lux, irradiation 16 hours, 30 days rear blades launch to form test-tube plantlet, and the test-tube plantlet inductivity can reach 100%.
2. the fast breeding of test-tube plantlet is cultivated: when the horseradish test-tube plantlet grows to the 1.5-2.0cm size, cut young shoot and be inoculated in differential medium, differential medium is MS+6-BA0.2mg/l+KT1.0mg/l+NAA.0.05mg/l+GA
30.05mg/l+ sucrose 4%, pH6.0, agar 0.65%, condition of culture is the same, and after 30 days, each young shoot can break up about the 4-5 strain, young shoot is downcut again, and change identical medium over to and repeat to cultivate, a large amount of breedings, ropagation coefficient of plantlet can reach 4.0-5.0.
3. test-tube plantlet strong seedling culture: through continuous culture after some generations, the test-tube plantlet growing way weakens sometimes, broke up prosperously, and change the strong seedling culture base over to after at this moment can will the test-tube plantlet young shoot downcutting, the strong seedling culture base is MS+6-BA0.2 mg/l+KT0.5mg/l+NAA0.05mg/l+GA
30.1mg/l+ sucrose 3%.Promote the blade petiole to stretch, cultivate 1-2 after generation, change above-mentioned proliferated culture medium again over to, promote its differentiation.
4. culture of rootage: the test-tube plantlet of enrichment culture, cut its young shoot, be inoculated on the root media, the culture of rootage based formulas is a MS+NAA0.1mg/l+ sucrose 2%, cultivation temperature is 18 ℃, illumination 1500Lux, and base portion can grow white adventive root 4-5 bar after 20 days, and more sturdy, rooting rate can reach more than 90%.
5. domestication is transplanted: the test-tube seedling transplanting front opening bottle cap of taking root carries out one week of adaptive training, take out then and rinse out medium, transplant in the nutritive cube that the sterilization vermiculite is housed, water from below, make moisture infiltrate the vermiculite surface, keep soil humidity, the control air themperature is at the 20-25 degree, can grow new root about 30 days and survive, survival rate can reach more than 85%.After test-tube plantlet survives, take out and transplant in the field, transplant the test-tube plantlet that domestication survives, transplanting survival rate can reach more than 80%.
Embodiment 2:
1. inducing of aseptic seedling: cut horseradish kind " true wife's " terminal bud, be cut into the 0.5cm size under the aseptic condition, be inoculated in inducing culture, the inducing culture based formulas is B
5+ 6-BA0.5mg/l+NAA0.1mg/l+GA
30.1mg/l+ sucrose 3%+ agar powder 0.6%, pH6.0, condition of culture be, 20 ℃ of temperature, and illumination 1500Lux, irradiation 14 hours, 30 days rear blades launch to form test-tube plantlet, and the test-tube plantlet inductivity can reach 100%.
2. the fast breeding of test-tube plantlet is cultivated: when the horseradish test-tube plantlet grows to the 1.5-2.0cm size, cut young shoot and be inoculated in differential medium, differential medium is B
5+ 6-BA0.5mg/l+KT0.5mg/l+NAA0.05mg/l+GA
30.1mg/l+ sucrose 4%+ agar 0.7%, pH6.0, condition of culture is the same, and after 30 days, each young shoot can break up about the 4-5 strain, young shoot is downcut again, and change identical medium over to and repeat to cultivate, a large amount of breedings, ropagation coefficient of plantlet can reach 4.0-5.0.
3. culture of rootage: the test-tube plantlet of enrichment culture, cut its young shoot, be inoculated on the root media, the culture of rootage based formulas is B
5+ IBA0.1mg/l+ sucrose 2%, cultivation temperature are 18 ℃, illumination 1500Lux, and base portion can grow the elongated adventive root 4-5 bar of white after 20 days, and rooting rate can reach 92%.
4. domestication is transplanted: open bottle cap before the test-tube seedling transplanting of taking root earlier, the method of taking partly to take off half lid prevents living contaminants, perform physical exercise a week, take out then and rinse out medium, transplant in the nutritive cube that fresh vermiculite is housed, can grow new root about 25 days and survive, survival rate can reach more than 90%.After test-tube plantlet survives, to transplant in the field, survival rate can reach more than 90%.
Embodiment 3:
1. test-tube plantlet is induced: get the terminal bud of horseradish kind " Yi Ze Bodhidharma ", sterile-processed after, be inoculated in following inducing culture: 1/2MS+6-BA0.4mg/l+NAA0.05mg/l+GA
30.1mg/l+ sucrose 3%+ agar powder 0.65%; 1/2MS+6-BA0.8mg/l+NAA0.1mg/l+GA
30.1mg/l+ sucrose 3%+ agar powder 0.65%, pH6.0, condition of culture be, 20 ℃ of temperature, and illumination 1500Lux, irradiation 16 hours, left and right sides mounted blade formed test-tube plantlet in 30 days, and the test-tube plantlet inductivity all can reach 100%.
2. the fast breeding of test-tube plantlet is cultivated: when the horseradish test-tube plantlet grows to the 1.5-2.0cm size, cut young shoot and be inoculated in following differential medium, differential medium is: MS+6-BA0.2mg/l+KT1.0mg/l+NAA0.2m/l+GA
30.05mg/l+ sucrose 4%, pH6. 0, agar 0
65%, reproduction coefficient is 5.0; MS+6-BA0.3mg/l+KT1.5mg/l+NAA0.1mg/l+GA
30.1mg/l+ sucrose 4%, reproduction coefficient are 4.5; MS+6-BA0.5mg/l+KT0.5mg/l+NAA0.05mg/l+GA
30.05mg/l+ sucrose 4%, reproduction coefficient are 4.3; MS+6-BA1.0mg/l+KT0.2mg/l+NAA0.2mg/l+GA
30.1mg/l+ fructose 5%, reproduction coefficient are 4.6; MS+6-BA1.5mg/l+KT0.3mg/l+NAA0.2mg/l+GA
30.3mg/l+ sucrose 3%, reproduction coefficient are 4.0; MS+6-BA0.8mg/l+KT0.5mg/l+NAA0.1mg/l+GA
30.1mg/l+ glucose 5%, reproduction coefficient are 4.2; MS+6-BA2.0mg/l+KT0.1mg/l+NAA0.3mg/l+GA
30.4mg/l+ sucrose 3%, reproduction coefficient are 4,5; MS+6-BA1.2mg/l+KT0.4mg/l+NAA0.2mg/l+GA
30.2mg/l+ sucrose 4%, reproduction coefficient are 4.8; MS+6-BA0.4mg/l+KT0.8mg/l+IAA0.1mg/+GA
30.1mg/l+ sucrose 4%, reproduction coefficient are 4.3; MS+6-BA0.3mg/l+KTl.0mg/l+IBA0.1mg/l+GA
30.1mg/l+ sucrose 4%, reproduction coefficient are 4.6; MS+6-BA0.1mg/l+KT2.0mg/l+IBA0.05mg/l+GA
30.2mg/l+ sucrose 3%, reproduction coefficient are 4.7; MS+6-BA0.4mg/l+KT0.3mg/l+IAA0.1mg/l+GA
30.1mg/l+ sucrose 3%, reproduction coefficient are 4.1; MS+6-BA0.2mg/l+KT0.4mg/l+IBA0.05mg/l+GA
30.05mg/l+ sucrose 5%, reproduction coefficient are 4.8.
Above condition of culture is all with aforementioned.
3. test-tube plantlet strong seedling culture: through continuous culture after some generations, the test-tube plantlet growing way weakens sometimes, broke up prosperously, and change the strong seedling culture base over to after at this moment can will the test-tube plantlet young shoot downcutting, the strong seedling culture base is MS+6-BA0.1mg/L+KT0.2mg/L+NAA0.05mg/L+GA
30.1mg/L+ sucrose 3%.Promote the blade petiole to stretch, cultivate 1-2 after generation, change above-mentioned proliferated culture medium again over to, promote its differentiation.
4. culture of rootage: the test-tube plantlet of enrichment culture, cut its young shoot, be inoculated on the root media, the culture of rootage based formulas is a MS+IAA0.05mg/L+ sucrose 2%, and cultivation temperature is 18 ℃, and illumination 1500Lux, rooting rate are 88%; MS+IAA0.1mmg/L+ sucrose 2%, rooting rate are 90%; MS+IBA0.05mg/L+ sucrose 2%, rooting rate are 91%.
5. domestication is transplanted: the test-tube seedling transplanting front opening bottle cap of taking root carries out one week of adaptive training, takes out then and washes medium off, transplants in the nutritive cube that the sterilization vermiculite is housed, and can grow new root about 30 days and survive, and survival rate can reach more than 85%.Test-tube plantlet takes out and transplants in the field, and transplanting survival rate can reach more than 80%.
Claims (5)
- Claims1. the method for quickly breeding of a horseradish seedling, it is characterized in that: 1) aseptic seedling is induced: cut the lateral bud on horseradish terminal bud or the root-like stock under the aseptic condition, be inoculated in MS or B 5+ 6-BA0.1-1mg/l+NAA0.05mg/l+GA 30.05-0.2mg/l+ carbohydrate 2-3% differentiates young shoot after 3～5 weeks of inoculation; 2) test-tube plantlet propagation: change test-tube plantlet over to MS (or B 5)+6-BA0.1-2mg/l+KT0.1-2mg/l+NAA0.01-0.5mg/l or IBA0.01-0.5mg/l or IAA0.01-0.5mg/l+GA 30.01-0.5mg/l on the proliferated culture medium, add carbohydrate 3-5%, inoculation back young shoot blade base axillalry bud is sprouted gradually, the switching back can form bud clump, shoot proliferation in 25-35 days; 3) culture of rootage: the no offspring of enrichment culture is inoculated in MS (or B after cutting 5On the root media of)+NAA0.01-0.2mg/l (or IBA0.01-0.2mg/l or IAA0.01-0.2mg/l)+carbohydrate 2-3%, taking root through 15-25 days seedling base portions becomes whole plant; 4) agar content is 0.6-0.8% in above-mentioned each medium, pH5.8-6.2.
- 2. according to the method for quickly breeding of the described horseradish seedling of claim 1, wherein each stage condition of culture is that cultivation temperature is 18-22 ℃, intensity of illumination 1500-2000Lux, irradiation 14-16 hour/day.
- 3. according to the method for quickly breeding of the described horseradish seedling of claim 1, in the shoot proliferation process, increase test-tube plantlet strong seedling culture process, change test-tube plantlet over to strong seedling culture base: MS (or B 5)+6-BA0.1-1mg/l+KT0.1-1mg/l+NAA0.05mg/l+GA 30.1-0.5mg/l+ carbohydrate 3-4% promotes the blade petiole to stretch, and changes above-mentioned proliferated culture medium again over to, promotes its differentiation.
- 4. according to the method for quickly breeding of the described horseradish seedling of claim 1, wherein used carbohydrate is meant sucrose, glucose, fructose etc., is best with sucrose.
- 5. according to the method for quickly breeding of the described horseradish seedling of claim 3, wherein used carbohydrate is meant sucrose, glucose, fructose etc., is best with sucrose.
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| CN98103484A CN1084141C (en) | 1998-08-05 | 1998-08-05 | Method for quick reproducing 'Shankui' sprout |
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| CN98103484A CN1084141C (en) | 1998-08-05 | 1998-08-05 | Method for quick reproducing 'Shankui' sprout |
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| CN1204453A true CN1204453A (en) | 1999-01-13 |
| CN1084141C CN1084141C (en) | 2002-05-08 |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102668992A (en) * | 2012-06-28 | 2012-09-19 | 井冈山大学 | Method for rooting of horseradish outside tissue culture seedling bottle |
| CN103053426A (en) * | 2013-02-04 | 2013-04-24 | 四川蓝公府农业科技有限公司 | Horseradish plantlet culture method |
| CN103053427A (en) * | 2013-02-04 | 2013-04-24 | 四川蓝公府农业科技有限公司 | Germination accelerating and stem tip sterilizing primary culture method for horseradish seeds |
| CN103081608A (en) * | 2013-02-26 | 2013-05-08 | 成都大学 | Germination processing method for horseradish seeds |
| CN103125386A (en) * | 2013-02-04 | 2013-06-05 | 四川蓝公府农业科技有限公司 | Industrial horseradish planting method |
| CN103718963A (en) * | 2013-12-24 | 2014-04-16 | 成都大学 | Culture method of virus-free seedling of horseradish |
| CN106358826A (en) * | 2016-08-30 | 2017-02-01 | 四川聚峰谷农业科技开发有限公司 | Cultivation method of myrciaria cauliflora |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH01262731A (en) * | 1988-04-13 | 1989-10-19 | Nippon Mining Co Ltd | Regeneration of plant body of shoot anlage |
| JPH05130815A (en) * | 1991-02-20 | 1993-05-28 | Katakura Kogyo Kk | Method for multiplying young seedling of wasabi and medium used therefor |
| JP2632631B2 (en) * | 1992-08-17 | 1997-07-23 | 株式会社ジャパンエナジー | Wasabi culture method |
| JP2957102B2 (en) * | 1994-12-02 | 1999-10-04 | ハウス食品株式会社 | Enlargement culture method of wasabi rhizome |
| JPH1094340A (en) * | 1996-09-20 | 1998-04-14 | Mitsubishi Materials Corp | Efficient propagation and rooting method for wasabi (japanese horseradish) |
-
1998
- 1998-08-05 CN CN98103484A patent/CN1084141C/en not_active Expired - Fee Related
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102668992A (en) * | 2012-06-28 | 2012-09-19 | 井冈山大学 | Method for rooting of horseradish outside tissue culture seedling bottle |
| CN103053426A (en) * | 2013-02-04 | 2013-04-24 | 四川蓝公府农业科技有限公司 | Horseradish plantlet culture method |
| CN103053427A (en) * | 2013-02-04 | 2013-04-24 | 四川蓝公府农业科技有限公司 | Germination accelerating and stem tip sterilizing primary culture method for horseradish seeds |
| CN103125386A (en) * | 2013-02-04 | 2013-06-05 | 四川蓝公府农业科技有限公司 | Industrial horseradish planting method |
| CN103053427B (en) * | 2013-02-04 | 2015-02-11 | 四川蓝公府农业科技有限公司 | Stem tip sterilizing primary culture method for germination accelerating for horseradish seeds |
| CN103125386B (en) * | 2013-02-04 | 2015-03-18 | 四川蓝公府农业科技有限公司 | Industrial horseradish planting method |
| CN103081608A (en) * | 2013-02-26 | 2013-05-08 | 成都大学 | Germination processing method for horseradish seeds |
| CN103081608B (en) * | 2013-02-26 | 2014-11-12 | 成都大学 | Germination processing method for horseradish seeds |
| CN103718963A (en) * | 2013-12-24 | 2014-04-16 | 成都大学 | Culture method of virus-free seedling of horseradish |
| CN103718963B (en) * | 2013-12-24 | 2015-07-29 | 成都大学 | A kind of cultural method of horseradish detoxic seedling |
| CN106358826A (en) * | 2016-08-30 | 2017-02-01 | 四川聚峰谷农业科技开发有限公司 | Cultivation method of myrciaria cauliflora |
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| CN1084141C (en) | 2002-05-08 |
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