CN103636501A - Tissue culture and rapid propagation method of Jinjili crabapple - Google Patents
Tissue culture and rapid propagation method of Jinjili crabapple Download PDFInfo
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- CN103636501A CN103636501A CN201310659036.6A CN201310659036A CN103636501A CN 103636501 A CN103636501 A CN 103636501A CN 201310659036 A CN201310659036 A CN 201310659036A CN 103636501 A CN103636501 A CN 103636501A
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Abstract
The invention relates to a tissue culture and rapid propagation method jinjili crabapple. Jinjili crabapple can be obtained through explant selection, explant cleaning, explant disinfection, primary culture, subculture and rooting culture. The tissue culture seedlings of jinjili crabapple obtained by the tissue culture method disclosed by the invention are high in rooting rate and survival speed; the propagation speed is high, the propagation coefficient is more than 5.3, and the rooting rate is over 90 percent; the obtained jinjili crabapple grows well and has a wide market prospect.
Description
Technical field
The present invention relates to field of plant tissue culture technique, refer to particularly a kind of Jin Jili Malus spectabilis quick breeding method for tissue culture.
Background technology
Jin Jili Malus spectabilis, is the rose family Malus griggles that introduce North America, and female male parent is not quite clear.Defoliation small arbor, leaf look glossy, spring, spire was aubergine, after transfer green to, pattern is red gorgeous, flower amount is large, bunchiness and giving birth to.Bloomed March in Wuhan Area at the beginning of 4 months, the florescence reaches two weeks, and young fruit is peony, and the later stage transfers cerise to, and result quantities is large, on branch, arranges successively, and the ability of being annual fruiting is strong, and the phase of bearing fruit is long, through processing, can be suspended to the Second Year January; Can do street tree, be also to see one of fine work seeds in fruit bowl scape, is the fine tree species of seeing flower, sight leaf, seeing fruit.
The Jin Jili Malus spectabilis seminal propagation cycle is long, resistance is poor, many employing propagation by grafiting now, but owing to being subject to the restriction of Quantity of the scion, can not carry out large-scale breeding, thus limited applying of this kind.Take tissue culture method to carry out the Fast-propagation of Jin Jili Malus spectabilis, set up axillary bud tissue and cultivate fast traditional font system, be conducive to Quick for a large amount of seedlings and be not subject to seasonal restrictions, Jin Jili is moved towards to Landscape Application and there is stronger production application and be worth.
Summary of the invention
The object of the invention is the defect existing for existing propagation technique, a kind of Jin Jili Malus spectabilis quick breeding method for tissue culture is provided.
For solving the problems of the technologies described above, a kind of Jin Jili Malus spectabilis quick breeding method for tissue culture provided by the invention, comprises the following steps:
1) explant is selected and is cleaned: the current-year branch that contains axillalry bud from the healthy and strong branch collection of the maternal plant of Jin Jili Malus spectabilis between 4～May, with suds, soak 5～15min, and standby after flowing water flushing 30～90min;
2) explant sterilization: the alcohol that is first 65～75% by volume fraction on superclean bench is by the explant sterilization 30～60s cleaning up, aseptic water washing 3～5 times, each 2～4min; Again with 0.2～0.3% mercuric chloride sterilization 5～10min, aseptic water washing 3～5 times, each 2～4min;
3) first culture: on superclean bench, the explant of disinfecting is cut into and contains the stem section that axillalry bud and length are 2～4cm, and by stem section access bud inducing culture, cultivation temperature is 25 ± 1, and ℃ light application time is 10～20h, light intensity 1800～2200Lx; Wherein, the formula of described bud inducing culture is: in MS medium+and 0.5～1.2mgL
-16-BA+0.05～0.3mgL
-1nAA+1～5% sucrose+0.2～1.0% agar, pH5.2～5.5.
4) subculture is cultivated: just in culture base, cultivating 20～25d, axillary bud sprouting grows Multiple Buds, and further growth becomes the first half of young tender plant, take mode separated between bud to be divided into independent spray the clump bud of 1.0～3.0cm left and right, transfer into proliferated culture medium, wherein, the formula of described proliferated culture medium is: in MS medium+and 1.5～3.0mgL
-16-BA+0.0.5～0.2mgL
-1nAA+2～6% sucrose+0.2～1.0% agar, pH5.2～5.5.
5) culture of rootage: in proliferated culture medium, cultivate 22～28d, the base portion clump bud of regenerating, the spray that length is surpassed to 2.5cm is separated from bud clump, and access root media is cultivated after 50d, forms the healthy and strong seedling of taking root; Wherein, the formula of described root media is: in MS medium+and 0.1～0.3mgL
-1iBA+2～6% sucrose+0.2～1.0% agar, pH5.2～5.5.
Further, Jin Jili Malus spectabilis quick breeding method for tissue culture, comprises the following steps:
1) explant is selected and is cleaned: the current-year branch that contains axillalry bud from the healthy and strong branch collection of the maternal plant of Jin Jili Malus spectabilis between 4～May, first with suds, soak 10min, and standby after flowing water flushing 60min;
2) explant sterilization: on superclean bench first with 70% alcohol by the explant sterilization 30s cleaning up, aseptic water washing 4 times, at every turn 2min; Again with 0.2% mercuric chloride sterilization 8min, aseptic water washing 5 times, each 2min;
3) first culture: on superclean bench, the explant of disinfecting is cut into and contains the stem section that 1 axillalry bud and length are 2cm, and by stem section access bud inducing culture, cultivation temperature is 25 ± 1, ℃ light application time is 15h, light intensity 1800～2200Lx, wherein, the formula of described bud inducing culture is: in MS medium+and 1.0mgL
-16-BA+0.2mgL
-1nAA+3% sucrose+0.54% agar, pH5.2～5.5.
4) subculture is cultivated: just in culture base, cultivating 20～25d, axillary bud sprouting grows Multiple Buds, and further growth becomes the first half of young tender plant, take mode separated between bud to be divided into independent spray the clump bud of 1.5cm, transfer into proliferated culture medium, wherein, the formula of described proliferated culture medium is: in MS medium+and 2.0mgL
-16-BA+0.1mgL
-1nAA+3% sucrose+0.54% agar, pH5.2～5.5.
5) culture of rootage: in proliferated culture medium, cultivate 22～28d, the base portion clump bud of regenerating, the spray that length is surpassed to 2.5cm is separated from bud clump, access root media, cultivates after 50d, forms the healthy and strong seedling of taking root, wherein, the formula of described root media is: in MS medium+and 0.2mgL
-1iBA+3% sucrose+0.54% agar, pH5.2～5.5.
Beneficial effect of the present invention is:
1, the breeding of Jin Jili Malus spectabilis realizes the anniversary: compared by the restriction of time with propagation by grafiting, the present invention can realize anniversary breeding in group training chamber, batch production is produced.
2, adopt that the Jin Jili Malus spectabilis group training seedling rooting rate that method for tissue culture of the present invention obtains is high, survival rate is high, reproduction speed is fast, and reproduction coefficient reaches more than 5.3, and rooting rate reaches more than 90%, the Jin Jili Malus spectabilis robust growth obtaining, has wide market prospects.
Embodiment
In order to explain better the present invention, below in conjunction with specific embodiment, further illustrate main contents of the present invention, but content of the present invention is not only confined to following examples.
Embodiment 1:
A Jin Jili Malus spectabilis quick breeding method for tissue culture, comprises the following steps:
1) explant is selected and is cleaned: the current-year branch that contains axillalry bud from the healthy and strong branch collection of the maternal plant of Jin Jili Malus spectabilis between 4～May, with suds, soak 5～15min, and standby after flowing water flushing 30～90min;
2) explant sterilization: the alcohol that is first 65～75% by volume fraction on superclean bench is by the explant sterilization 30～60s cleaning up, aseptic water washing 3～5 times, each 2～4min; Again with 0.2～0.3% mercuric chloride sterilization 5～10min, aseptic water washing 3～5 times, each 2～4min;
3) first culture: on superclean bench, the explant of disinfecting is cut into and contains the stem section that axillalry bud and length are 2～4cm, and by stem section access bud inducing culture, cultivation temperature is 25 ± 1, and ℃ light application time is 10～20h, light intensity 1800～2200Lx; Wherein, the formula of described bud inducing culture is: in MS medium+and 0.5～1.2mgL
-16-BA+0.05～0.3mgL
-1nAA+1～5% sucrose+0.2～1.0% agar, pH5.2～5.5.
4) subculture is cultivated: just in culture base, cultivating 20～25d, axillary bud sprouting grows Multiple Buds, and further growth becomes the first half of young tender plant, take mode separated between bud to be divided into independent spray the clump bud of 1.0～3.0cm left and right, transfer into proliferated culture medium, wherein, the formula of described proliferated culture medium is: in MS medium+and 1.5～3.0mgL
-16-BA+0.0.5～0.2mgL
-1nAA+2～6% sucrose+0.2～1.0% agar, pH5.2～5.5.
5) culture of rootage: in proliferated culture medium, cultivate 22～28d, the base portion clump bud of regenerating, the spray that length is surpassed to 2.5cm is separated from bud clump, and access root media is cultivated after 50d, forms the healthy and strong seedling of taking root; Wherein, the formula of described root media is: in MS medium+and 0.1～0.3mgL
-1iBA+2～6% sucrose+0.2～1.0% agar, pH5.2～5.5.
Embodiment 2
Jin Jili Malus spectabilis quick breeding method for tissue culture, comprises the following steps:
1) explant is selected and is cleaned: the current-year branch that contains axillalry bud from the healthy and strong branch collection of the maternal plant of Jin Jili Malus spectabilis between 4～May, first with suds, soak 10min, and standby after flowing water flushing 60min;
2) explant sterilization: on superclean bench first with 70% alcohol by the explant sterilization 30s cleaning up, aseptic water washing 4 times, at every turn 2min; Again with 0.2% mercuric chloride sterilization 8min, aseptic water washing 5 times, each 2min;
3) first culture: on superclean bench, the explant of disinfecting is cut into and contains the stem section that 1 axillalry bud and length are 2cm, and by stem section access bud inducing culture, cultivation temperature is 25 ± 1, ℃ light application time is 15h, light intensity 1800～2200Lx, wherein, the formula of described bud inducing culture is: in MS medium+and 1.0mgL
-16-BA+0.2mgL
-1nAA+3% sucrose+0.54% agar, pH5.2～5.5.
4) subculture is cultivated: just in culture base, cultivating 20～25d, axillary bud sprouting grows Multiple Buds, and further growth becomes the first half of young tender plant, take mode separated between bud to be divided into independent spray the clump bud of 1.5cm, transfer into proliferated culture medium, wherein, the formula of described proliferated culture medium is: in MS medium+and 2.0mgL
-16-BA+0.1mgL
-1nAA+3% sucrose+0.54% agar, pH5.2～5.5.
5) culture of rootage: in proliferated culture medium, cultivate 22～28d, the base portion clump bud of regenerating, the spray that length is surpassed to 2.5cm is separated from bud clump, access root media, cultivates after 50d, forms the healthy and strong seedling of taking root, wherein, the formula of described root media is: in MS medium+and 0.2mgL
-1iBA+3% sucrose+0.54% agar, pH5.2～5.5.
Other unspecified part is prior art.Although above-described embodiment has been made detailed description to the present invention; but it is only the present invention's part embodiment; rather than whole embodiment, people can also obtain other embodiment according to the present embodiment under without creative prerequisite, and these embodiment belong to protection domain of the present invention.
Claims (2)
1. Yi Zhong Jin Jili Malus spectabilis quick breeding method for tissue culture, is characterized in that: comprise the following steps:
1) explant is selected and is cleaned: the current-year branch that 4-5 contains axillalry bud from the healthy and strong branch collection of the maternal plant of Jin Jili Malus spectabilis between the month, with suds, soak 5～15min, and standby after flowing water flushing 30～90min;
2) explant sterilization: the alcohol that is first 65～75% by volume fraction on superclean bench is by the explant sterilization 30～60s cleaning up, aseptic water washing 3～5 times, each 2～4min; Again with 0.2～0.3% mercuric chloride sterilization 5～10min, aseptic water washing 3～5 times, each 2～4min;
3) first culture: on superclean bench, the explant of disinfecting is cut into and contains the stem section that axillalry bud and length are 2～4cm, and by stem section access bud inducing culture, cultivation temperature is 25 ± 1, and ℃ light application time is 10～20h, light intensity 1800～2200Lx; Wherein, the formula of described bud inducing culture is: in MS medium+and 0.5～1.2mgL
-16-BA+0.05～0.3mgL
-1nAA+1～5% sucrose+0.2～1.0% agar, pH5.2～5.5;
4) subculture is cultivated: just in culture base, cultivating 20～25d, axillary bud sprouting grows Multiple Buds, and further growth becomes the first half of young tender plant, take mode separated between bud to be divided into independent spray the clump bud of 1.0～3.0cm left and right, transfer into proliferated culture medium, wherein, the formula of described proliferated culture medium is: in MS medium+and 1.5～3.0mgL
-16-BA+0.0.5～0.2mgL
-1nAA+2～6% sucrose+0.2～1.0% agar, pH5.2～5.5;
5) culture of rootage: in proliferated culture medium, cultivate 22～28d, the base portion clump bud of regenerating, the spray that length is surpassed to 2.5cm is separated from bud clump, and access root media is cultivated after 50d, forms the healthy and strong seedling of taking root; Wherein, the formula of described root media is: in MS medium+and 0.1～0.3mgL
-1iBA+2～6% sucrose+0.2～1.0% agar, pH5.2～5.5.
2. Jin Jili Malus spectabilis quick breeding method for tissue culture according to claim 1, is characterized in that: comprise the following steps:
1) explant is selected and is cleaned: the current-year branch that contains axillalry bud from the healthy and strong branch collection of the maternal plant of Jin Jili Malus spectabilis between 4～May, first with suds, soak 10min, and standby after flowing water flushing 60min;
2) explant sterilization: on superclean bench first with 70% alcohol by the explant sterilization 30s cleaning up, aseptic water washing 4 times, at every turn 2min; Again with 0.2% mercuric chloride sterilization 8min, aseptic water washing 5 times, each 2min;
3) first culture: on superclean bench, the explant of disinfecting is cut into and contains the stem section that 1 axillalry bud and length are 2cm, and by stem section access bud inducing culture, cultivation temperature is 25 ± 1, ℃ light application time is 15h, light intensity 1800～2200Lx, wherein, the formula of described bud inducing culture is: in MS medium+and 1.0mgL
-16-BA+0.2mgL
-1nAA+3% sucrose+0.54% agar, pH5.2～5.5;
4) subculture is cultivated: just in culture base, cultivating 20～25d, axillary bud sprouting grows Multiple Buds, and further growth becomes the first half of young tender plant, take mode separated between bud to be divided into independent spray the clump bud of 1.5cm, transfer into proliferated culture medium, wherein, the formula of described proliferated culture medium is: in MS medium+and 2.0mgL
-16-BA+0.1mgL
-1nAA+3% sucrose+0.54% agar, pH5.2～5.5;
5) culture of rootage: in proliferated culture medium, cultivate 22～28d, the base portion clump bud of regenerating, the spray that length is surpassed to 2.5cm is separated from bud clump, access root media, cultivates after 50d, forms the healthy and strong seedling of taking root, wherein, the formula of described root media is: in MS medium+and 0.2mgL
-1iBA+3% sucrose+0.54% agar, pH5.2～5.5.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104585036A (en) * | 2015-01-29 | 2015-05-06 | 南京林业大学 | Tissue culture rapid-propagation method of fire axillary bud of North American crabapple tableland |
| CN106212274A (en) * | 2016-07-15 | 2016-12-14 | 武汉市农业科学技术研究院林业果树科学研究所 | The method for tissue culture of special beautiful Caulis et folium euphorbiae milii |
| CN107637520A (en) * | 2017-09-27 | 2018-01-30 | 南京林业大学 | A kind of method for tissue culture of Hubei Chinese flowering crabapple |
| CN109601380A (en) * | 2018-12-26 | 2019-04-12 | 武汉市农业科学院 | A kind of tissue culture and rapid propagation method of the dance Malus spectabilis of fire |
| CN115956504A (en) * | 2023-01-06 | 2023-04-14 | 北京农学院 | Primary culture method for crabapple plants |
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| CN101352149A (en) * | 2008-09-02 | 2009-01-28 | 南京林业大学 | Method for quickly breeding Malus 'Hongying' by tissue culture |
| RU2485768C1 (en) * | 2012-03-20 | 2013-06-27 | Государственное научное учреждение Всероссийский научно-исследовательский институт садоводства им. И.В. Мичурина Россельхозакадемии (ГНУ ВНИИС им. И.В. Мичурина РАСХН) | Nutrient medium for rhizogenesis of apple tree and pear tree in vitro |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| JPH0322934A (en) * | 1989-06-19 | 1991-01-31 | Takeda Chem Ind Ltd | Production of plant seedling |
| CN101352149A (en) * | 2008-09-02 | 2009-01-28 | 南京林业大学 | Method for quickly breeding Malus 'Hongying' by tissue culture |
| RU2485768C1 (en) * | 2012-03-20 | 2013-06-27 | Государственное научное учреждение Всероссийский научно-исследовательский институт садоводства им. И.В. Мичурина Россельхозакадемии (ГНУ ВНИИС им. И.В. Мичурина РАСХН) | Nutrient medium for rhizogenesis of apple tree and pear tree in vitro |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104585036A (en) * | 2015-01-29 | 2015-05-06 | 南京林业大学 | Tissue culture rapid-propagation method of fire axillary bud of North American crabapple tableland |
| CN106212274A (en) * | 2016-07-15 | 2016-12-14 | 武汉市农业科学技术研究院林业果树科学研究所 | The method for tissue culture of special beautiful Caulis et folium euphorbiae milii |
| CN107637520A (en) * | 2017-09-27 | 2018-01-30 | 南京林业大学 | A kind of method for tissue culture of Hubei Chinese flowering crabapple |
| CN109601380A (en) * | 2018-12-26 | 2019-04-12 | 武汉市农业科学院 | A kind of tissue culture and rapid propagation method of the dance Malus spectabilis of fire |
| CN115956504A (en) * | 2023-01-06 | 2023-04-14 | 北京农学院 | Primary culture method for crabapple plants |
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