CN105010144A - Double cherry tissue culture propagation method - Google Patents

Double cherry tissue culture propagation method Download PDF

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Publication number
CN105010144A
CN105010144A CN201510448520.3A CN201510448520A CN105010144A CN 105010144 A CN105010144 A CN 105010144A CN 201510448520 A CN201510448520 A CN 201510448520A CN 105010144 A CN105010144 A CN 105010144A
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China
Prior art keywords
culture
mgl
illumination
tissue culture
explant
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Pending
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CN201510448520.3A
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Chinese (zh)
Inventor
樊仲书
曾毓
张玉华
陈天增
樊昌华
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Fujian Century Jinhua Technology Co Ltd
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Fujian Century Jinhua Technology Co Ltd
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Priority to CN201510448520.3A priority Critical patent/CN105010144A/en
Publication of CN105010144A publication Critical patent/CN105010144A/en
Pending legal-status Critical Current

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Abstract

The invention provides a double cherry tissue culture propagation method. An economical efficient double cherry tissue culture propagation technology system is established according to the steps of explant selection, explant sterilization, tissue culture seedling induction, tissue culture seedling proliferation and tissue culture seedling rooting. An improved MS culture medium is adopted, the purposes of improving the double cherry transplantation survival rate, ensuring high propagation coefficient and short period and realizing rapid-propagation seedling culture without external environment influence can be achieved, the excellent characteristics of excellent varieties can be preserved, and the problem in short supply of double cherry is alleviated.

Description

A kind of eightfold cherry tissue culture and rapid propagation method
Technical field
The present invention relates to a kind of tissue culture and rapid propagation method, particularly relate to a kind of eightfold cherry tissue culture and rapid propagation method, be applicable to the rapid seedling cultivation of eightfold cherry.
Background technology
Oriental cherry has very high ornamental value, and flower is charming delicate and charming, completely sets splendid as rosy clouds, and therefore whenever oriental cherry blooms the time, the Sacura Festival all can be held in various places; Oriental cherry also has the effect of skin care, shows according to the study, and oriental cherry can pore refining, Sage Extract and lightening skin color, containing abundant natural vitamin A, B, E, is often extracted and makes skin care item; But the life of oriental cherry is very of short duration, an oriental cherry is approximately 7 days from opening into wither, and whole cherry tree from blooming entirely thanking approximately just about 16 days, cannot meet the demand of viewing and admiring of people and the research and development of all kinds of skin-protection product.
Eightfold cherry belongs to rosaceous plant, is deciduous tree, and another name " red cherry " is one of them classification of oriental cherry, leaf egg shape lanceolar, long 8-12 centimetre, ora serrata, alternate; Flower is large, and peony, grows thickly for several, and pendency is open, and petal is multiple and close, the light fragrance of tool, and bennet is long and smooth; Growth characteristics: like sunlight and the environment that cools, the 3-4 month blooms; Primary growth is in Japan, wherein famous with Nara eightfold cherry, and current China various places also start plantation, as garden landscape, view and admire for people; Eightfold cherry is DoubleCherry, and seed is rare, and traditional propagation method seedling is slow, is difficult to a large amount of production; And adopt group culturation rapid propagating technology to be a kind of approach efficiently for the numerous soon of improved seeds, organize according to made oriental cherry and trained successfully, but the tissue culture technology of eightfold cherry kind, current efficiency is desirable not enough.
Summary of the invention
The object of the invention is to provide a kind of eightfold cherry tissue culture and rapid propagation method to overcome above-mentioned deficiency, improve the transplanting survival rate of eightfold cherry, reproduction coefficient is high, cycle is short, be not affected by the external environment, both kept the good characteristic of improved seeds, alleviate again the predicament that supply falls short of demand of eightfold cherry seedling.
For achieving the above object, the present invention carries out in the following way.
A kind of eightfold cherry tissue culture and rapid propagation method, the step of taking root according to explant selection, explant sterilization, plantlet in vitro induction, plantlet in vitro propagation, plantlet in vitro is carried out.
Being chosen as of described explant is chosen sunny day in spring clip in the morning and is newly sent out branch and be in the tender shoots fringe bar of semi-lignified state as explant, draws materials conveniently, easily.
The sterilization of described explant and the way of plantlet in vitro Fiber differentiation are that the explant chosen is soaked 20min with in saturated bleaching powder supernatant, constantly stir in immersion process, then rinse with clear water, dry with filter paper, be placed on superclean bench, first use the alcohol immersion 15s of 75%, then explant is placed in 0.1% mercuric chloride dilution and soaks 8min, then use aseptic water washing 6 times, be then inoculated on inducing culture, inducing culture is additional 1.5 mgL of modified MS medium -16-BA, 0.1 mgL -1nAA, sucrose 30 gL -1with carragheen 6 gL -1, be placed in constant temperature biochemical cultivation case and cultivate; Inducing culturing condition is, temperature 25 ± 2 DEG C, illumination 12/d, intensity of illumination 1000LX.
Described plantlet in vitro Multiplying culture step is after Fiber differentiation 30d, cuts single for indefinite bud, is inoculated on the proliferated culture medium that has been equipped with, and proliferated culture medium is additional 1.2 mgL of modified MS medium -16-BA, 0.1 mgL -1nAA, sucrose 30 gL -1with carragheen 6 gL -1; In incubation, observe its growth phenomenon, after 30d, its growth coefficient is 3.0; Multiplying culture condition is, temperature 25 ± 2 DEG C, illumination 12/d, intensity of illumination 2000LX.
Described plantlet in vitro culture of rootage is after Multiplying culture 30d, selects stalwartness and simple bud height 1.5-2.5cm, is inoculated on the root media that has been equipped with, and root media is additional 0.5 mgL of 1/2 modified MS medium -1i-BA, 0.2 mgL -1nAA, sucrose 30 gL -1with carragheen 6 gL -1), in incubation, observe its growth phenomenon, after 30d, its rooting rate is 95%, and height of seedling reaches 2.5-3.5cm, plantation of can emerging; Culture of rootage condition is, temperature 25 ± 2 DEG C, illumination 12/d, intensity of illumination 2000LX.
The formula of above-described modified MS medium is.
A kind of eightfold cherry tissue culture and rapid propagation method provided by the invention, according to the step that explant selection, explant sterilization, plantlet in vitro induction, plantlet in vitro propagation, plantlet in vitro are taken root, establish the fast numerous tissue culture technology system of eightfold cherry of a set of economical and efficient, have employed modified MS medium, reach the transplanting survival rate that improve eightfold cherry, reach the object of rapid seedling cultivation.
This rapid seedling cultivation technology and conventional art contrast and are listed as follows:
Embodiment
Under regard to specific embodiments of the invention and elaborate further.
A kind of eightfold cherry tissue culture and rapid propagation method, is characterized in that the method comprises the following steps.
(1) selection of explant: sunny day in spring clip in the morning is newly sent out branch and is in the tender shoots fringe bar of semi-lignified state as explant.
(2) sterilization of explant and plantlet in vitro Fiber differentiation: the tender shoots fringe bar cut from parent is soaked 20min with in saturated bleaching powder supernatant, constantly stir in immersion process, fringe bar water after immersion rinses, then dry with filter paper, be placed on superclean bench, first use the alcohol immersion 15s of 75%, then explant is placed in 0.1% mercuric chloride and soaks 8min, then use aseptic water washing 6 times, be then inoculated on inducing culture, inducing culture is additional 1.5 mgL of modified MS medium -16-BA, 0.1 mgL -1nAA, sucrose 30 gL -1with carragheen 6 gL -1, be placed in constant temperature biochemical cultivation case and cultivate; Inducing culturing condition is, temperature 25 ± 2 DEG C, illumination 12/d, intensity of illumination 1000LX.
(3) plantlet in vitro Multiplying culture: after Fiber differentiation 30d, cut single for indefinite bud, be inoculated on the proliferated culture medium that has been equipped with, proliferated culture medium is the additional 1.2 mgL-1 6-BA of modified MS medium, 0.1 mgL-1 NAA, sucrose 30 gL-1 and carragheen 6 gL-1; In incubation, observe its growth phenomenon, after 30d, its growth coefficient is 3.0; Multiplying culture condition is, temperature 25 ± 2 DEG C, illumination 12/d, intensity of illumination 2000LX.
(4) plantlet in vitro culture of rootage: after Multiplying culture 30d, select stalwartness and simple bud height 1.5-2.5cm, be inoculated on the root media that has been equipped with, root media is 1/2 modified MS medium additional 0.5 mgL-1 I-BA, 0.2 mgL-1 NAA, sucrose 30 gL-1 and carragheen 6 gL-1), in incubation, observe its growth phenomenon, after 30d, its rooting rate is 95%, height of seedling reaches 2.5-3.5cm, plantation of can emerging; Culture of rootage condition is, temperature 25 ± 2 DEG C, illumination 12/d, intensity of illumination 2000LX.
Above-described embodiment is only the present invention's preferably embodiment; but embodiments of the present invention are not restricted to the described embodiments; other changes made under not deviating from Spirit Essence of the present invention and principle, substitute, combine; all should be the substitute mode of equivalence, be included in protection scope of the present invention.

Claims (2)

1. an eightfold cherry tissue culture and rapid propagation method, is characterized in that the method comprises the following steps:
(1) selection of explant: sunny day in spring clip in the morning is newly sent out branch and is in the tender shoots fringe bar of semi-lignified state as explant;
(2) sterilization of explant and plantlet in vitro Fiber differentiation: the tender shoots fringe bar cut from parent is soaked 20min with in saturated bleaching powder supernatant, constantly stir in immersion process, fringe bar water after immersion rinses, then dry with filter paper, be placed on superclean bench, first use the alcohol immersion 15s of 75%, then explant is placed in 0.1% mercuric chloride and soaks 8min, then aseptic water washing is used 6 times, then be inoculated on inducing culture, inducing culture is the additional 1.5 mgL-1 6-BA of modified MS medium, 0.1 mgL-1 NAA, sucrose 30 gL-1 and carragheen 6 gL-1, be placed in constant temperature biochemical cultivation case to cultivate, inducing culturing condition is, temperature 25 ± 2 DEG C, illumination 12/d, intensity of illumination 1000LX,
(3) plantlet in vitro Multiplying culture: after Fiber differentiation 30d, cut single for indefinite bud, be inoculated on the proliferated culture medium that has been equipped with, proliferated culture medium is the additional 1.2 mgL-1 6-BA of modified MS medium, 0.1 mgL-1 NAA, sucrose 30 gL-1 and carragheen 6 gL-1; In incubation, observe its growth phenomenon, after 30d, its growth coefficient is 3.0; Multiplying culture condition is, temperature 25 ± 2 DEG C, illumination 12/d, intensity of illumination 2000LX;
(4) plantlet in vitro culture of rootage: after Multiplying culture 30d, select stalwartness and simple bud height 1.5-2.5cm, be inoculated on the root media that has been equipped with, root media is 1/2 modified MS medium additional 0.5 mgL-1 I-BA, 0.2 mgL-1 NAA, sucrose 30 gL-1 and carragheen 6 gL-1), in incubation, observe its growth phenomenon, after 30d, its rooting rate is 95%, height of seedling reaches 2.5-3.5cm, plantation of can emerging; Culture of rootage condition is, temperature 25 ± 2 DEG C, illumination 12/d, intensity of illumination 2000LX.
2. a kind of eightfold cherry tissue culture and rapid propagation method according to claim 1, is characterized in that described modified MS medium formula is .
CN201510448520.3A 2015-07-28 2015-07-28 Double cherry tissue culture propagation method Pending CN105010144A (en)

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105340742A (en) * 2015-11-16 2016-02-24 广州天适集团有限公司 Tissue culture rapid propagation method for cerasus yunnanensis(Franch.)Yu et Li adult excellent single plant 'Guangzhou' cerasus yunnanensis
CN105379621A (en) * 2015-11-16 2016-03-09 华南农业大学 Efficient in-vitro plant regeneration method of adult high-quality single-plant Xiaoqiao oriental cherry of cerasus lannesiana var. speciosa
CN105494100A (en) * 2015-12-30 2016-04-20 江汉大学 Tissue culture and rapid propagation method of oriental cherries
CN107135943A (en) * 2017-04-20 2017-09-08 广州旺地园林工程有限公司 A kind of winter cherry rapid propagation in vitro method
CN107509634A (en) * 2017-09-30 2017-12-26 绍兴文理学院元培学院 A kind of gold thread grass tissue cultures and rapid propagation method
CN111837946A (en) * 2019-09-30 2020-10-30 宁波城市职业技术学院 Efficient oriental cherry tissue culture rapid propagation method

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CN101584299A (en) * 2009-04-15 2009-11-25 河南省农业科学院 A kind of red autumnal leaves oriental cherry tissue-culturing rapid propagation medium and tissue culture and rapid propagation method

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JP2003274777A (en) * 2002-03-22 2003-09-30 Meiji Univ Method for transferring gene to plant by using agrobacterium
CN1541516A (en) * 2003-11-06 2004-11-03 中国科学院遗传与发育生物学研究所 Tissue cultivation quick breeding method of grass cherry blossom
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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105340742A (en) * 2015-11-16 2016-02-24 广州天适集团有限公司 Tissue culture rapid propagation method for cerasus yunnanensis(Franch.)Yu et Li adult excellent single plant 'Guangzhou' cerasus yunnanensis
CN105379621A (en) * 2015-11-16 2016-03-09 华南农业大学 Efficient in-vitro plant regeneration method of adult high-quality single-plant Xiaoqiao oriental cherry of cerasus lannesiana var. speciosa
CN105379621B (en) * 2015-11-16 2017-12-29 华南农业大学 A kind of high-efficiency in-vitro plant regeneration method of Prunus donarium adult fine individual plant " little Qiao " cherry
CN105494100A (en) * 2015-12-30 2016-04-20 江汉大学 Tissue culture and rapid propagation method of oriental cherries
CN107135943A (en) * 2017-04-20 2017-09-08 广州旺地园林工程有限公司 A kind of winter cherry rapid propagation in vitro method
CN107509634A (en) * 2017-09-30 2017-12-26 绍兴文理学院元培学院 A kind of gold thread grass tissue cultures and rapid propagation method
CN111837946A (en) * 2019-09-30 2020-10-30 宁波城市职业技术学院 Efficient oriental cherry tissue culture rapid propagation method

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