CN107410019A - A kind of method that sweet potato tissue cultures are carried out using classification culture medium - Google Patents

A kind of method that sweet potato tissue cultures are carried out using classification culture medium Download PDF

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Publication number
CN107410019A
CN107410019A CN201710198013.8A CN201710198013A CN107410019A CN 107410019 A CN107410019 A CN 107410019A CN 201710198013 A CN201710198013 A CN 201710198013A CN 107410019 A CN107410019 A CN 107410019A
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China
Prior art keywords
sweet potato
test tube
culture
tube seedling
culture medium
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CN201710198013.8A
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Chinese (zh)
Inventor
王立国
王刚
李燕
李建军
高翔
张嘉
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Tianjin Fenghua Yulong Agricultural Development Co Ltd
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Tianjin Fenghua Yulong Agricultural Development Co Ltd
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Priority to CN201710198013.8A priority Critical patent/CN107410019A/en
Publication of CN107410019A publication Critical patent/CN107410019A/en
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    • 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
    • A01H4/001—Culture apparatus for tissue culture
    • 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

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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)

Abstract

The present invention is a kind of method for carrying out sweet potato tissue cultures using classification culture medium, is concretely comprised the following steps：The foundation of explant；The induction of callus：Inducing culture is MS+KT 2mg/L+2,4 D 0.5mg/L or MS+KT 2mg/L+NAA 0.5mg/L；The differentiation of callus：Differential medium is MS+KT 1mg/L+BA 1mg/L+IAA 0.5mg/L+GA30.1mg/L or MS+KT 2mg/L+NAA 0.5mg/L；The propagation of test tube seedling and growth：Proliferated culture medium is MS；Test tube seedling is taken root：Root media is 1/2MS；The transplanting of rooting tube plantlet.Tissue cultures are carried out to sweet potato present invention employs classification culture medium, is commercially produced for the fast numerous and extensive test tube seedling of sweet potato and provides scientific basis.

Description

A kind of method that sweet potato tissue cultures are carried out using classification culture medium
Technical field
The present invention relates to the field of sweet potato tissue cultures, more particularly to a kind of use to be classified culture medium progress sweet potato tissue training Foster method.
Background technology
Sweet potato is one of four big staple food crop of China, and feed and the important source material of light industry.As China gives birth to The development of production and the change of people's eating patterns, sweet potato is as work(such as the collection energy, food processing, health care and medicinal, foreign exchange earnings Industrial crops that can be, have been to be concerned by more and more people.Sweet potato is a kind of hybrid vigour crop, but uses and nourish and generate Virus Diseases of Sweet Potato its spread in china can often be caused, cause yield and Quality Down.
The content of the invention
Present invention seek to address that the deficiencies in the prior art, and provide a kind of using classification culture medium progress sweet potato tissue cultures Method.
The present invention to achieve the above object, using following technical scheme：
A kind of method for carrying out sweet potato tissue cultures using classification culture medium, is concretely comprised the following steps：
(1) foundation of explant：
Take Sweet Potato Leaf to be rinsed with running water, 5-10min is then soaked in washing powder solution, then rinsed and done with running water Only, it is placed on super-clean bench and carries out disinfection, the blockage that blade is cut into 0.5cm × 0.5cm after the completion of sterilization is standby；
(2) induction of callus：
It is enterprising by inducing culture MS+KT 2mg/L+2,4-D0.5mg/L are inoculated in by the Sweet Potato Leaf disinfected Row light culture 15 days；Or inducing culture MS+KT 2mg/L+NAA will be inoculated in by the Sweet Potato Leaf disinfected Optical culture is carried out on 0.5mg/L 12 days；
(3) differentiation of callus：
The callus being inoculated on inducing culture MS+KT 2mg/L+2,4-D 0.5mg/L is transferred and cultivated in differentiation Base MS+KT 1mg/L+BA 1mg/L+IAA 0.5mg/L+GA3Optical culture is carried out on 0.1mg/L 45 days；Or it will be inoculated in and lure Lead callus on culture medium MS+KT 2mg/L+NAA 0.5mg/L and directly carry out optical culture 45 days；
(4) propagation of test tube seedling and growth：
The test tube seedling that differentiates will be induced to be cut into the stem section of 1 axillary bud of band to be forwarded on MS proliferated culture mediums and carry out propagation life It is long；
(5) test tube seedling is taken root：
Test tube seedling by propagation growth is inoculated on 1/2MS root medias and taken root；
(6) transplanting of rooting tube plantlet：
When test tube seedling root long 1.2cm, the natural light lower refining seedling 2 days in greenhouse, then open sealed membrane 3 days, with sterile washing Root culture medium is removed, is transplanted into greenhouse, heat and moisture preserving growth.
During the foundation of explant, the process that Sweet Potato Leaf carries out disinfection on super-clean bench is：Sweet Potato Leaf is existed Soaked in 75% alcohol, soak time 30-40s, with aseptic water washing 5-8 times；Then with 1% HgCl2Solution is carried out 15min is sterilized, with aseptic water washing 5-8 times；Finally blade surface moisture is blotted with aseptic paper.
In the rooting process of test tube seedling, root media 1/2MS pH is 5.8-6.0, agar 0.7%, while using 3% White sugar and running water.
The beneficial effects of the invention are as follows：Tissue cultures are carried out to sweet potato present invention employs classification culture medium, instead of tradition Nourish and generate, so can as far as possible avoid virus breeding；Different phase is cultivated using different culture mediums, is that sweet potato is fast Numerous and extensive test tube seedling, which is commercially produced, provides scientific basis.
Embodiment
With reference to specific embodiment, the invention will be further described：
Embodiment 1：
A kind of method for carrying out sweet potato tissue cultures using classification culture medium, is concretely comprised the following steps：
(1) foundation of explant：
Take Sweet Potato Leaf to be rinsed with running water, 5min then soaked in washing powder solution, then rinsed well with running water, It is placed on super-clean bench and carries out disinfection, the blockage that blade is cut into 0.5cm × 0.5cm after the completion of sterilization is standby；
(2) induction of callus：
It is enterprising by inducing culture MS+KT 2mg/L+2,4-D0.5mg/L are inoculated in by the Sweet Potato Leaf disinfected Row light culture 15 days；
(3) differentiation of callus：
The callus being inoculated on inducing culture MS+KT 2mg/L+2,4-D 0.5mg/L is transferred and cultivated in differentiation Base MS+KT 1mg/L+BA 1mg/L+IAA 0.5mg/L+GA3Optical culture is carried out on 0.1mg/L 45 days；
(4) propagation of test tube seedling and growth：
The test tube seedling that differentiates will be induced to be cut into the stem section of 1 axillary bud of band to be forwarded on MS proliferated culture mediums and carry out propagation life It is long；
(5) test tube seedling is taken root：
Test tube seedling by propagation growth is inoculated on 1/2MS root medias and taken root；
(6) transplanting of rooting tube plantlet：
When test tube seedling root long 1.2cm, the natural light lower refining seedling 2 days in greenhouse, then open sealed membrane 3 days, with sterile washing Root culture medium is removed, is transplanted into greenhouse, heat and moisture preserving growth.
During the foundation of explant, the process that Sweet Potato Leaf carries out disinfection on super-clean bench is：Sweet Potato Leaf is existed Soaked in 75% alcohol, soak time 30s, with aseptic water washing 5 times；Then with 1% HgCl2Solution carries out disinfection 15min, with aseptic water washing 5 times；Finally blade surface moisture is blotted with aseptic paper.
In the rooting process of test tube seedling, root media 1/2MS pH is 5.8, agar 0.7%, while uses 3% white sugar And running water.
Embodiment 2：
A kind of method for carrying out sweet potato tissue cultures using classification culture medium, is concretely comprised the following steps：
(1) foundation of explant：
Take Sweet Potato Leaf to be rinsed with running water, 10min is then soaked in washing powder solution, then rinsed and done with running water Only, it is placed on super-clean bench and carries out disinfection, the blockage that blade is cut into 0.5cm × 0.5cm after the completion of sterilization is standby；
(2) induction of callus：
Carried out being inoculated in by the Sweet Potato Leaf disinfected on inducing culture MS+KT 2mg/L+NAA0.5mg/L Optical culture 12 days；
(3) differentiation of callus：
Callus on inducing culture MS+KT 2mg/L+NAA 0.5mg/L will be inoculated in and directly carry out optical culture 45 My god；
(4) propagation of test tube seedling and growth：
The test tube seedling that differentiates will be induced to be cut into the stem section of 1 axillary bud of band to be forwarded on MS proliferated culture mediums and carry out propagation life It is long；
(5) test tube seedling is taken root：
Test tube seedling by propagation growth is inoculated on 1/2MS root medias and taken root；
(6) transplanting of rooting tube plantlet：
When test tube seedling root long 1.2cm, the natural light lower refining seedling 2 days in greenhouse, then open sealed membrane 3 days, with sterile washing Root culture medium is removed, is transplanted into greenhouse, heat and moisture preserving growth.
During the foundation of explant, the process that Sweet Potato Leaf carries out disinfection on super-clean bench is：Sweet Potato Leaf is existed Soaked in 75% alcohol, soak time 40s, with aseptic water washing 8 times；Then with 1% HgCl2Solution carries out disinfection 15min, with aseptic water washing 8 times；Finally blade surface moisture is blotted with aseptic paper.
In the rooting process of test tube seedling, root media 1/2MS pH is 6.0, agar 0.7%, while uses 3% white sugar And running water.
Embodiment 3：
A kind of method for carrying out sweet potato tissue cultures using classification culture medium, is concretely comprised the following steps：
(1) foundation of explant：
Take Sweet Potato Leaf to be rinsed with running water, 8min then soaked in washing powder solution, then rinsed well with running water, It is placed on super-clean bench and carries out disinfection, the blockage that blade is cut into 0.5cm × 0.5cm after the completion of sterilization is standby；
(2) induction of callus：
It is enterprising by inducing culture MS+KT 2mg/L+2,4-D0.5mg/L are inoculated in by the Sweet Potato Leaf disinfected Row light culture 15 days；
(3) differentiation of callus：
The callus being inoculated on inducing culture MS+KT 2mg/L+2,4-D 0.5mg/L is transferred and cultivated in differentiation Base MS+KT 1mg/L+BA 1mg/L+IAA 0.5mg/L+GA3Optical culture is carried out on 0.1mg/L 45 days；
(4) propagation of test tube seedling and growth：
The test tube seedling that differentiates will be induced to be cut into the stem section of 1 axillary bud of band to be forwarded on MS proliferated culture mediums and carry out propagation life It is long；
(5) test tube seedling is taken root：
Test tube seedling by propagation growth is inoculated on 1/2MS root medias and taken root；
(6) transplanting of rooting tube plantlet：
When test tube seedling root long 1.2cm, the natural light lower refining seedling 2 days in greenhouse, then open sealed membrane 3 days, with sterile washing Root culture medium is removed, is transplanted into greenhouse, heat and moisture preserving growth.
During the foundation of explant, the process that Sweet Potato Leaf carries out disinfection on super-clean bench is：Sweet Potato Leaf is existed Soaked in 75% alcohol, soak time 35s, with aseptic water washing 6 times；Then with 1% HgCl2Solution carries out disinfection 15min, with aseptic water washing 7 times；Finally blade surface moisture is blotted with aseptic paper.
In the rooting process of test tube seedling, root media 1/2MS pH is 5.9, agar 0.7%, while uses 3% white sugar And running water.
Tissue cultures are carried out to sweet potato present invention employs classification culture medium, nourishes and generates instead of traditional, so can Virus breeding is avoided as far as possible；Different phase is cultivated using different culture mediums, is the fast numerous and extensive test tube seedling business of sweet potato The production of industry metaplasia provides scientific basis.
The present invention is exemplarily described above in conjunction with specific embodiment, it is clear that present invention specific implementation is not by upper The limitation of mode is stated, if the various improvement of inventive concept and technical scheme of the present invention progress are employed, or it is not improved straight Scoop out for other occasions, within protection scope of the present invention.

Claims (3)

  1. A kind of 1. method that sweet potato tissue cultures are carried out using classification culture medium, it is characterised in that concretely comprise the following steps：
    (1) foundation of explant：
    Take Sweet Potato Leaf to be rinsed with running water, 5-10min then soaked in washing powder solution, then rinsed well with running water, It is placed on super-clean bench and carries out disinfection, the blockage that blade is cut into 0.5cm × 0.5cm after the completion of sterilization is standby；
    (2) induction of callus：
    It will be inoculated on inducing culture MS+KT 2mg/L+2,4-D0.5mg/L and be carried out secretly by the Sweet Potato Leaf disinfected Culture 15 days；Or inducing culture MS+KT 2mg/L+NAA 0.5mg/L will be inoculated in by the Sweet Potato Leaf disinfected Upper progress optical culture 12 days；
    (3) differentiation of callus：
    The callus being inoculated on inducing culture MS+KT 2mg/L+2,4-D 0.5mg/L is transferred in differential medium MS +KT 1mg/L+BA 1mg/L+IAA 0.5mg/L+GA3Optical culture is carried out on 0.1mg/L 45 days；Or induction training will be inoculated in Support callus on base MS+KT 2mg/L+NAA 0.5mg/L and directly carry out optical culture 45 days；
    (4) propagation of test tube seedling and growth：
    The test tube seedling that differentiates will be induced to be cut into the stem section of 1 axillary bud of band to be forwarded on MS proliferated culture mediums and carry out propagation growth；
    (5) test tube seedling is taken root：
    Test tube seedling by propagation growth is inoculated on 1/2MS root medias and taken root；
    (6) transplanting of rooting tube plantlet：
    When test tube seedling root long 1.2cm, the natural light lower refining seedling 2 days in greenhouse, then open sealed membrane 3 days, wash away root with sterilized water Portion's culture medium, is transplanted into greenhouse, heat and moisture preserving growth.
  2. A kind of 2. method that sweet potato tissue cultures are carried out using classification culture medium according to claim 1, it is characterised in that During the foundation of explant, the process that Sweet Potato Leaf carries out disinfection on super-clean bench is：By alcohol of the Sweet Potato Leaf 75% Middle immersion, soak time 30-40s, with aseptic water washing 5-8 times；Then with 1% HgCl2Solution carries out disinfection 15min, With aseptic water washing 5-8 times；Finally blade surface moisture is blotted with aseptic paper.
  3. A kind of 3. method that sweet potato tissue cultures are carried out using classification culture medium according to claim 2, it is characterised in that In the rooting process of test tube seedling, root media 1/2MS pH is 5.8-6.0, agar 0.7%, while 3% white sugar of use and oneself Water.
CN201710198013.8A 2017-03-29 2017-03-29 A kind of method that sweet potato tissue cultures are carried out using classification culture medium Pending CN107410019A (en)

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CN201710198013.8A CN107410019A (en) 2017-03-29 2017-03-29 A kind of method that sweet potato tissue cultures are carried out using classification culture medium

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108432764A (en) * 2018-04-03 2018-08-24 洛阳超拓园林绿化工程有限公司 A kind of root-growing agent improving palisades hole plant setting survival rate
CN116686714A (en) * 2023-07-11 2023-09-05 泰安市农业科学院 Quick preparation and long-term preservation method of sweet potato germplasm resource test-tube plantlet

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108432764A (en) * 2018-04-03 2018-08-24 洛阳超拓园林绿化工程有限公司 A kind of root-growing agent improving palisades hole plant setting survival rate
CN116686714A (en) * 2023-07-11 2023-09-05 泰安市农业科学院 Quick preparation and long-term preservation method of sweet potato germplasm resource test-tube plantlet

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