CN108901846A - A kind of preparation manipulation technology of simple quick Sweetpotato Viruses Elimination seedling - Google Patents
A kind of preparation manipulation technology of simple quick Sweetpotato Viruses Elimination seedling Download PDFInfo
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- CN108901846A CN108901846A CN201810762399.5A CN201810762399A CN108901846A CN 108901846 A CN108901846 A CN 108901846A CN 201810762399 A CN201810762399 A CN 201810762399A CN 108901846 A CN108901846 A CN 108901846A
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- seedling
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- callus
- detoxic
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- 244000017020 Ipomoea batatas Species 0.000 title claims abstract description 16
- 235000002678 Ipomoea batatas Nutrition 0.000 title claims abstract description 16
- 241000700605 Viruses Species 0.000 title claims abstract description 13
- 230000008030 elimination Effects 0.000 title claims abstract description 13
- 238000003379 elimination reaction Methods 0.000 title claims abstract description 13
- 238000005516 engineering process Methods 0.000 title claims abstract description 13
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- 206010020649 Hyperkeratosis Diseases 0.000 claims abstract description 15
- 239000001963 growth medium Substances 0.000 claims abstract description 14
- 241000196324 Embryophyta Species 0.000 claims description 22
- 238000005286 illumination Methods 0.000 claims description 12
- 241000238631 Hexapoda Species 0.000 claims description 9
- 239000003337 fertilizer Substances 0.000 claims description 9
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 6
- 239000004202 carbamide Substances 0.000 claims description 6
- 238000012258 culturing Methods 0.000 claims description 6
- 230000002265 prevention Effects 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 230000000249 desinfective effect Effects 0.000 claims description 4
- 238000001784 detoxification Methods 0.000 claims description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 3
- 241000921313 Phyllopodium Species 0.000 claims description 3
- 244000061456 Solanum tuberosum Species 0.000 claims description 3
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 3
- 241000607479 Yersinia pestis Species 0.000 claims description 3
- 230000015572 biosynthetic process Effects 0.000 claims description 3
- TWFZGCMQGLPBSX-UHFFFAOYSA-N carbendazim Chemical compound C1=CC=C2NC(NC(=O)OC)=NC2=C1 TWFZGCMQGLPBSX-UHFFFAOYSA-N 0.000 claims description 3
- 150000001875 compounds Chemical class 0.000 claims description 3
- 238000003745 diagnosis Methods 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 230000000763 evoking effect Effects 0.000 claims description 3
- 238000011534 incubation Methods 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 229960002523 mercuric chloride Drugs 0.000 claims description 3
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims description 3
- 230000000474 nursing effect Effects 0.000 claims description 3
- 238000009400 out breeding Methods 0.000 claims description 3
- 239000008223 sterile water Substances 0.000 claims description 3
- 230000003612 virological effect Effects 0.000 claims description 3
- 239000004563 wettable powder Substances 0.000 claims description 3
- 238000009395 breeding Methods 0.000 abstract description 6
- 230000001488 breeding effect Effects 0.000 abstract description 6
- 239000000463 material Substances 0.000 abstract description 6
- 238000004659 sterilization and disinfection Methods 0.000 abstract description 5
- 230000004069 differentiation Effects 0.000 abstract description 4
- 230000006698 induction Effects 0.000 abstract description 3
- 230000001939 inductive effect Effects 0.000 abstract description 3
- 239000002609 medium Substances 0.000 abstract description 3
- 238000000034 method Methods 0.000 description 8
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
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- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a kind of preparation manipulation technologies of simple quick Sweetpotato Viruses Elimination seedling, and the threshold of the technology is low, does not need anatomical lens and professional technician to remove stem apex, general skilled worker and most basic tissue culture room condition can be operated；The stem apex for using aseptic seedling, does not need sterilisation step, as long as suitable operation, will not pollute substantially；Simplify step, stem apex can be initially formed callus, regenerated root, then seedling after being inoculated into inductive differentiation medium, not need repeatedly to turn culture medium；This technology is also easy to produce mutative material by callus induction seedling, provides new breeding material for breeder and carries out breed breeding.
Description
Technical field
The present invention relates to Sweetpotato Viruses Elimination seedling technical field more particularly to a kind of preparation manipulations of simple quick Sweetpotato Viruses Elimination seedling
Technology.
Background technique
Original stem apex stripping means requires technical strong, it is desirable that 1, the method for stem apex disinfection and time will hold essence
Standard, disinfecting time is too long, and meeting damaging tissue, too short, disinfection is not thorough, and pollutes.2, it needs to operate under anatomical lens, such as
The fruit operating time is too long, then the stem-tip tissue removed may be dead.3, the stem apex removed is it is not possible that very detoxification is clean, still
It so needs to detect by molecule and serum.And the horizontal proportional example phase of operating technology of virus elimination rate success rate and different technologies personnel
It closes.4, original poison-removing method needs to turn 2-3 bottle, could obtain intact plant（Such as Fiber differentiation ,-differentiation is cultivated-takes root
Culture）Workload is bigger.
Summary of the invention
The purpose of the present invention is to solve disadvantages existing in the prior art, and a kind of simple quick sweet potato proposed is de-
The preparation manipulation technology of malicious seedling.
To achieve the goals above, present invention employs following technical solutions：
A kind of preparation manipulation technology of simple quick Sweetpotato Viruses Elimination seedling, which is characterized in that include the following steps,
S1, vernalization：The undamaged sweet potato potato wedge of free from insect pests is chosen, impregnates 15- with 50% carbendazol wettable powder, 800 times of liquid
20min is put into 35 DEG C of dark vernalization in incubator after drying；
S2, preparation aseptic seedling：When bud length to 10-15cm or so, the terminal bud of top 1-2cm long is intercepted, macroscopic leaf is cut off
Piece is first rinsed with clear water, then with 75% alcohol disinfecting 30s, then with 0.1% mercuric chloride solution is sterilized 6-8min, is finally used sterile water
Rinsing 3-5 times；It being inoculated in ms culture medium, cultivation temperature is 25-27 DEG C, daily illumination in 10 hours, and 3500lx is cultivated 3-4 weeks,
Complete aseptic plant can be obtained；
S3, detoxication and tissue culture：The terminal bud of aseptic plant is taken in super-clean bench, is removed blade, is left and taken phyllopodium position about 1*1*1mm3
The tissue of size.It is divided into 4-8 parts with scalpel cutting, is inoculated in and is added to 0.1-0.2mg/LNAA+1.0-2.0mg/L6-BA's
Dark culturing on culture medium, cultivation temperature are 25-27 DEG C, the formation of evoked callus.Callus is formed after 3 weeks, after 5 weeks
Callus is taken root, and goes to daily illumination in 8 hours, light intensity 3000-3500lx, is cultivated about 4 weeks at 25-27 DEG C, is obtained complete plant
Strain；
S4, each single plant of acquisition is numbered, carries out viral diagnosis, rejected non-detoxification completely and do not meet variety characteristic
Plant；
S5, detoxic seedling are numerous fastly：Detoxic seedling is cut into single-unit and is inoculated on MS+1.0-2.0mg/LGA3 culture medium, is in cultivation temperature
28-30 DEG C, under conditions of intensity of illumination is 3500lx, 10h illumination cultivation is carried out daily, it incubation time 35-40 days can subculture one
It is secondary, fast heavy multiple 5 steps；
The domestication of S6, detoxic seedling：When detoxic seedling is long to nearly position of bottleneck, move out culturing room, be placed in sunlight can not direct projection arrive
1 week or so at concealment, during which gradually opens bottle cap and tamed；
The transplanting of S7, detoxic seedling：Detoxic seedling is cleaned into culture medium, divides single plant plant in seedlings nursing plate.After slow seedling survives at concealment,
It is transplanted in insect prevention greenhouse；
After S8, greenhouse are transplanted 3 weeks, urea is spread fertilizer over the fields, is watered immediately, promotes growth of seedling；
S9, when vines are long to 30cm or more, shear vines plant in insect prevention greenhouse, carry out breeding again for detoxic seedling, cut every time
After climing, urea+compound fertilizer is spread fertilizer over the fields, is watered immediately, is i.e. acquisition Sweetpotato Viruses Elimination seedling.
Beneficial effects of the present invention：
1, the threshold of this method is low, does not need anatomical lens and professional technician to remove stem apex, general skilled worker and most
Basic tissue culture room condition can be operated；
2, the stem apex for using aseptic seedling, does not need sterilisation step, as long as suitable operation, will not pollute substantially；
3, simplify step, stem apex can be initially formed callus, regenerated root, then seedling after being inoculated into inductive differentiation medium, not need more
It is secondary to turn culture medium；
4, this method is also easy to produce mutative material by callus induction seedling, provides new breeding material for breeder and carries out kind
Breeding.
Specific embodiment
Technical scheme in the embodiment of the invention is clearly and completely described, it is clear that described embodiment is only
It is a part of the embodiment of the present invention, instead of all the embodiments.
A kind of preparation manipulation technology of simple quick Sweetpotato Viruses Elimination seedling, its step are as follows：
S1, vernalization：The undamaged sweet potato potato wedge of free from insect pests is chosen, impregnates 15- with 50% carbendazol wettable powder, 800 times of liquid
20min is put into 35 DEG C of dark vernalization in incubator after drying；
S2, preparation aseptic seedling：When bud length to 10-15cm or so, the terminal bud of top 1-2cm long is intercepted, macroscopic leaf is cut off
Piece is first rinsed with clear water, then with 75% alcohol disinfecting 30s, then with 0.1% mercuric chloride solution is sterilized 6-8min, is finally used sterile water
Rinsing 3-5 times；It being inoculated in ms culture medium, cultivation temperature is 25-27 DEG C, daily illumination in 10 hours, and 3500lx is cultivated 3-4 weeks,
Complete aseptic plant can be obtained；
S3, detoxication and tissue culture：The terminal bud of aseptic plant is taken in super-clean bench, is removed blade, is left and taken phyllopodium position about 1*1*1mm3
The tissue of size.It is divided into 4-8 parts with scalpel cutting, is inoculated in and is added to 0.1-0.2mg/LNAA+1.0-2.0mg/L6-BA's
Dark culturing on culture medium, cultivation temperature are 25-27 DEG C, the formation of evoked callus.Callus is formed after 3 weeks, after 5 weeks
Callus is taken root, and goes to daily illumination in 8 hours, light intensity 3000-3500lx, is cultivated about 4 weeks at 25-27 DEG C, is obtained complete plant
Strain；
S4, each single plant of acquisition is numbered, carries out viral diagnosis, rejected non-detoxification completely and do not meet variety characteristic
Plant；
S5, detoxic seedling are numerous fastly：Detoxic seedling is cut into single-unit and is inoculated on MS+1.0-2.0mg/LGA3 culture medium, is in cultivation temperature
28-30 DEG C, under conditions of intensity of illumination is 3500lx, 10h illumination cultivation is carried out daily, it incubation time 35-40 days can subculture one
It is secondary, fast heavy multiple 5 steps；
The domestication of S6, detoxic seedling：When detoxic seedling is long to nearly position of bottleneck, move out culturing room, be placed in sunlight can not direct projection arrive
1 week or so at concealment, during which gradually opens bottle cap and tamed；
The transplanting of S7, detoxic seedling：Detoxic seedling is cleaned into culture medium, divides single plant plant in seedlings nursing plate.After slow seedling survives at concealment,
It is transplanted in insect prevention greenhouse；
After S8, greenhouse are transplanted 3 weeks, urea is spread fertilizer over the fields, is watered immediately, promotes growth of seedling；
S9, when vines are long to 30cm or more, shear vines plant in insect prevention greenhouse, carry out breeding again for detoxic seedling, cut every time
After climing, urea+compound fertilizer is spread fertilizer over the fields, is watered immediately, is i.e. acquisition Sweetpotato Viruses Elimination seedling.
A kind of preparation manipulation technology of simple quick Sweetpotato Viruses Elimination seedling proposed by the present invention, the threshold of this method is low, is not required to
Anatomical lens and professional technician are wanted to remove stem apex, general skilled worker and most basic tissue culture room condition can be operated；
The stem apex for using aseptic seedling, does not need sterilisation step, as long as suitable operation, will not pollute substantially；Simplify step, stem apex connects
It can be initially formed callus, regenerated root, then seedling after kind to inductive differentiation medium, do not need repeatedly to turn culture medium；This method passes through
Callus induction seedling, is also easy to produce mutative material, provides new breeding material for breeder and carries out breed breeding.
The foregoing is only a preferred embodiment of the present invention, but scope of protection of the present invention is not limited thereto,
Anyone skilled in the art in the technical scope disclosed by the present invention, according to the technique and scheme of the present invention and its
Inventive concept is subject to equivalent substitution or change, should be covered by the protection scope of the present invention.
Claims (4)
1. a kind of preparation manipulation technology of simple quick Sweetpotato Viruses Elimination seedling, which is characterized in that include the following steps,
S1, vernalization：The undamaged sweet potato potato wedge of free from insect pests is chosen, impregnates 15- with 50% carbendazol wettable powder, 800 times of liquid
20min is put into 35 DEG C of dark vernalization in incubator after drying；
S2, preparation aseptic seedling：When bud length to 10-15cm or so, the terminal bud of top 1-2cm long is intercepted, macroscopic leaf is cut off
Piece is first rinsed with clear water, then with 75% alcohol disinfecting 30s, then with 0.1% mercuric chloride solution is sterilized 6-8min, is finally used sterile water
Rinsing 3-5 times；It being inoculated in ms culture medium, cultivation temperature is 25-27 DEG C, daily illumination in 10 hours, and 3500lx is cultivated 3-4 weeks,
Complete aseptic plant can be obtained；
S3, detoxication and tissue culture：The terminal bud of aseptic plant is taken in super-clean bench, is removed blade, is left and taken phyllopodium position about 1*1*1mm3Greatly
Small tissue.
2. being divided into 4-8 parts with scalpel cutting, it is inoculated in the culture for being added to 0.1-0.2mg/LNAA+1.0-2.0mg/L6-BA
Dark culturing on base, cultivation temperature are 25-27 DEG C, the formation of evoked callus.
Callus is formed after 3.3 weeks, callus is taken root after 5 weeks, go to daily illumination in 8 hours, light intensity 3000-3500lx,
It is cultivated at 25-27 DEG C about 4 weeks, obtains intact plant；
S4, each single plant of acquisition is numbered, carries out viral diagnosis, rejected non-detoxification completely and do not meet variety characteristic
Plant；
S5, detoxic seedling are numerous fastly：Detoxic seedling is cut into single-unit and is inoculated into MS+1.0-2.0mg/LGA3On culture medium, it is in cultivation temperature
28-30 DEG C, under conditions of intensity of illumination is 3500lx, 10h illumination cultivation is carried out daily, it incubation time 35-40 days can subculture one
It is secondary, fast heavy multiple 5 steps；
The domestication of S6, detoxic seedling：When detoxic seedling is long to nearly position of bottleneck, move out culturing room, be placed in sunlight can not direct projection arrive
1 week or so at concealment, during which gradually opens bottle cap and tamed；
The transplanting of S7, detoxic seedling：Detoxic seedling is cleaned into culture medium, divides single plant plant in seedlings nursing plate.
4. after slow seedling survives at concealment, being transplanted in insect prevention greenhouse；
After S8, greenhouse are transplanted 3 weeks, urea is spread fertilizer over the fields, is watered immediately, promotes growth of seedling；
S9, when vines are long to 30cm or more, shear vines plant in insect prevention greenhouse, carry out breeding again for detoxic seedling, cut every time
After climing, urea+compound fertilizer is spread fertilizer over the fields, is watered immediately, is i.e. acquisition Sweetpotato Viruses Elimination seedling.
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| CN201810762399.5A CN108901846A (en) | 2018-07-12 | 2018-07-12 | A kind of preparation manipulation technology of simple quick Sweetpotato Viruses Elimination seedling |
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| CN201810762399.5A CN108901846A (en) | 2018-07-12 | 2018-07-12 | A kind of preparation manipulation technology of simple quick Sweetpotato Viruses Elimination seedling |
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| CN201810762399.5A Pending CN108901846A (en) | 2018-07-12 | 2018-07-12 | A kind of preparation manipulation technology of simple quick Sweetpotato Viruses Elimination seedling |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109329027A (en) * | 2018-12-03 | 2019-02-15 | 湖南省作物研究所 | A kind of efficient propagation method of sweet potato |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101584301A (en) * | 2009-06-05 | 2009-11-25 | 海南省农业科学院粮食作物研究所 | The method of culturing detoxified seedling by sweet potato stem tip |
| CN104067821A (en) * | 2014-06-26 | 2014-10-01 | 青岛农业大学 | Preparation method of virus-free seedlings of sweet potato |
| CN104719164A (en) * | 2015-03-30 | 2015-06-24 | 青岛农业大学 | Rapid propagation method of virus-free breeder potato seeds of sweet potatoes |
| CN106258065A (en) * | 2016-08-03 | 2017-01-04 | 天津丰华裕隆农业发展有限公司 | The stem apex detoxification method for culturing seedlings of Autumn Gold Rhizoma Dioscoreae esculentae |
-
2018
- 2018-07-12 CN CN201810762399.5A patent/CN108901846A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101584301A (en) * | 2009-06-05 | 2009-11-25 | 海南省农业科学院粮食作物研究所 | The method of culturing detoxified seedling by sweet potato stem tip |
| CN104067821A (en) * | 2014-06-26 | 2014-10-01 | 青岛农业大学 | Preparation method of virus-free seedlings of sweet potato |
| CN104719164A (en) * | 2015-03-30 | 2015-06-24 | 青岛农业大学 | Rapid propagation method of virus-free breeder potato seeds of sweet potatoes |
| CN106258065A (en) * | 2016-08-03 | 2017-01-04 | 天津丰华裕隆农业发展有限公司 | The stem apex detoxification method for culturing seedlings of Autumn Gold Rhizoma Dioscoreae esculentae |
Non-Patent Citations (1)
| Title |
|---|
| 熊继文: "马铃薯脱毒种薯高产栽培技术", vol. 1, 30 April 2007, 贵州科技出版社, pages: 16 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109329027A (en) * | 2018-12-03 | 2019-02-15 | 湖南省作物研究所 | A kind of efficient propagation method of sweet potato |
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