CN102812905A - Blueberry tender stem tissue culturing and rapid propagation process - Google Patents
Blueberry tender stem tissue culturing and rapid propagation process Download PDFInfo
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- CN102812905A CN102812905A CN2012103402996A CN201210340299A CN102812905A CN 102812905 A CN102812905 A CN 102812905A CN 2012103402996 A CN2012103402996 A CN 2012103402996A CN 201210340299 A CN201210340299 A CN 201210340299A CN 102812905 A CN102812905 A CN 102812905A
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Abstract
The invention discloses a blueberry tender stem tissue culturing and rapid propagation process. The process comprises the following steps: S1, selecting tender stem without lignification from a field, purifying, disinfecting and cutting into stem sections with bud; S2, carrying out induction culturing, wherein the induction culture medium contains MS, 3.0mg/L of ZT (zeatin), 0.1mg/L of IBA (indolebutyric acid), 30g/L of sucrose and 6.5g/L of agar powder; and S3, carrying out subculturing, wherein the multiplication culture medium contains MS, 0.5-1.5mg/L of ZT, 0.05-0.1mg/L of IBA, 30g/L of sucrose and 6.5g/L of agar powder, and carrying out nursery stock rejuvenation by using the culture medium containing MS and0.3-0.7mg/L of ZT before transplanting of the blueberry. The process has the beneficial effects of high reproductive rate, high reproductive speed and low cost of tissue culture seedling; and the cultured nursery stock is high in quality and uniform, can remove virus easily, has developed root systems, and provides a favorable basis for the later-stage tree body growth result.
Description
Technical field
The present invention relates to blueberry tissue culture technique field, the tender stem segment tissue culture fast breeding technology of particularly a kind of blueberry.
Background technology
Blueberry belongs to Du magpie flower section Vaccinium shrub, is a kind of little berry fruit tree, and its fruit has very high nutritive value and economic worth.Existing market constantly increases the demand of blueberry, but the plant husbandry of China's blueberry is at the early-stage, and a large amount of improved seeds nursery stocks are badly in need of in various places when setting up the plantation.Adopt conventional hard branch, green wood cutting survival rate low, the cycle is long, and speed is slow, and is prone to poison in spite of illness, can't satisfy market to the growing needs of blueberry nursery stock quantity.
Summary of the invention
The objective of the invention is to overcome the shortcoming of prior art, a kind of reproduction rate height, the fast tender stem segment tissue culture fast breeding technology of blueberry of reproduction speed are provided.
The object of the invention is realized through following technical scheme: the tender stem segment tissue culture fast breeding technology of blueberry, and it may further comprise the steps:
S1, choose not lignified tender stem from the field, behind flushing with clean water 50～60min, on aseptic workbench; Clean 20～30min with the water that adds liquid detergent; Use 75% alcohol disinfecting, 20～30s then, aseptic water washing 3～5 times is afterwards on superclean bench; Put into 0.1% mercury chloride thimerosal, 8～10min, with aseptic water washing 2～3 times; On superclean bench, tender stem is cut into stem with bud then, removes the part branches and leaves;
S2, inducing culture: on superclean bench, the stem with bud that shears is inoculated in the cultivation of sprouting on the inducing culture, inducing culture is meant MS+ZT 3.0mg/L+IBA 0.1mg/L+ sucrose 30g/L+ agar powder 6.5g/L, and regulating its pH is 5.2～5.8;
S3, successive transfer culture: after treating to grow on the tender stem the long sprouting of 1cm; Sprouting is downcut; Be inoculated in and carry out inducing clumping bud and shoot proliferation on the shoot proliferation medium; Per 40～50 days is a successive transfer culture cycle, and the shoot proliferation medium is meant MS+ZT 0.5～1.5mg/L+IBA0.05～0.1mg/L+ sucrose 30g/L+ agar 6.5g/L, and regulating its pH is 5.2～5.8; 2～3 successive transfer culture adopt MS+ZT 0.3～0.7mg/L medium to carry out the nursery stock rejuvenation in the cycle before the blueberry bottle outlet is transplanted.
The present invention has the following advantages:
1, reproduction rate of the present invention is high, reproduction speed is fast, and it is with low cost to produce tissue cultivating seedling, and the tissue cultivating seedling heredity shape that obtains is consistent, has overcome the low shortcoming of conventional reproduction coefficient.
2, the seedling quality of the little numerous production of group training is good, and uniformity is sloughed virus easily, well developed root system, and tree bulk-growth result lays a good foundation for the later stage.
Embodiment
Below in conjunction with embodiment the present invention is done further description, but the invention is not restricted to be described below.
Embodiment 1:
The tender stem segment tissue culture fast breeding technology of blueberry, it may further comprise the steps:
S1, choose not lignified tender stem from the field, behind the flushing with clean water 50min, on aseptic workbench; Clean 20min with the water that adds liquid detergent; Use 75% alcohol disinfecting 30s then, aseptic water washing 5 times is afterwards on superclean bench; Put into 0.1% mercury chloride thimerosal 8min, with aseptic water washing 3 times; On superclean bench, tender stem is cut into stem with bud then, removes the part branches and leaves;
S2, inducing culture: on superclean bench, the stem with bud that shears is inoculated in the cultivation of sprouting on the inducing culture, inducing culture is meant MS+ZT 3.0mg/L+IBA 0.1mg/L+ sucrose 30g/L+ agar powder 6.5g/L, and regulating its pH is 5.4;
S3, successive transfer culture: after treating to grow on the tender stem the long sprouting of 1cm; Sprouting is downcut; Be inoculated in and carry out inducing clumping bud and shoot proliferation on the shoot proliferation medium; Per 40 days is a successive transfer culture cycle, and the shoot proliferation medium is meant MS+ZT1.5mg/L+IBA0.1mg/L+ sucrose 30g/L+ agar 6.5g/L, and regulating its pH is 5.4; 2 successive transfer culture adopt MS+ZT 0.7mg/L in the cycle before the blueberry bottle outlet is transplanted, and pH is that 5.4 medium carries out the nursery stock rejuvenation.
Embodiment 2:
The tender stem segment tissue culture fast breeding technology of blueberry, it may further comprise the steps:
S1, choose not lignified tender stem from the field, behind the flushing with clean water 60min, on aseptic workbench; Clean 30min with the water that adds liquid detergent; Use 75% alcohol disinfecting 20s then, aseptic water washing 3 times is afterwards on superclean bench; Put into 0.1% mercury chloride thimerosal 10min, with aseptic water washing 2 times; On superclean bench, tender stem is cut into stem with bud then, removes the part branches and leaves;
S2, inducing culture: on superclean bench, the stem with bud that shears is inoculated in the cultivation of sprouting on the inducing culture, inducing culture is meant MS+ZT 3.0mg/L+IBA 0.1mg/L+ sucrose 30g/L+ agar powder 6.5g/L, and regulating its pH is 5.2;
S3, successive transfer culture: after treating to grow on the tender stem the long sprouting of 1cm; Sprouting is downcut; Be inoculated in and carry out inducing clumping bud and shoot proliferation on the shoot proliferation medium; Per 50 days is a successive transfer culture cycle, and the shoot proliferation medium is meant MS+ZT 0.5mg/L+IBA0.05mg/L+ sucrose 30g/L+ agar 6.5g/L, and regulating its pH is 5.2; 3 successive transfer culture adopt MS+ZT 0.3mg/L in the cycle before the blueberry bottle outlet is transplanted, and pH is that 5.2 medium carries out the nursery stock rejuvenation.
Embodiment 3:
The tender stem segment tissue culture fast breeding technology of blueberry, it may further comprise the steps:
S1, choose not lignified tender stem from the field, behind the flushing with clean water 55min, on aseptic workbench; Clean 25min with the water that adds liquid detergent; Use 75% alcohol disinfecting 25s then, aseptic water washing 4 times is afterwards on superclean bench; Put into 0.1% mercury chloride thimerosal 9min, with aseptic water washing 3 times; On superclean bench, tender stem is cut into stem with bud then, removes the part branches and leaves;
S2, inducing culture: on superclean bench, the stem with bud that shears is inoculated in the cultivation of sprouting on the inducing culture, inducing culture is meant MS+ZT 3.0mg/L+IBA 0.1mg/L+ sucrose 30g/L+ agar powder 6.5g/L, and regulating its pH is 5.8;
S3, successive transfer culture: after treating to grow on the tender stem the long sprouting of 1cm; Sprouting is downcut; Be inoculated in and carry out inducing clumping bud and shoot proliferation on the shoot proliferation medium; Per 45 days is a successive transfer culture cycle, and the shoot proliferation medium is meant MS+ZT 0.5～1.5mg/L+IBA0.08mg/L+ sucrose 30g/L+ agar 6.5g/L, and regulating its pH is 5.8; 2～3 successive transfer culture adopt MS+ZT 0.3～0.7mg/L in the cycle before the blueberry bottle outlet is transplanted, and pH is that 5.8 medium carries out the nursery stock rejuvenation.
Claims (1)
1. the tender stem segment tissue culture fast breeding technology of blueberry, it is characterized in that: it may further comprise the steps:
S1, choose not lignified tender stem from the field, behind flushing with clean water 50～60min, on aseptic workbench; Clean 20～30min with the water that adds liquid detergent; Use 75% alcohol disinfecting, 20～30s then, aseptic water washing 3～5 times is afterwards on superclean bench; Put into 0.1% mercury chloride thimerosal, 8～10min, with aseptic water washing 2～3 times; On superclean bench, tender stem is cut into stem with bud then, removes the part branches and leaves;
S2, inducing culture: on superclean bench, the stem with bud that shears is inoculated in the cultivation of sprouting on the inducing culture, inducing culture is meant MS+ZT 3.0mg/L+IBA 0.1mg/L+ sucrose 30g/L+ agar powder 6.5g/L, and regulating its pH is 5.2～5.8;
S3, successive transfer culture: after treating to grow on the tender stem the long sprouting of 1cm; Sprouting is downcut; Be inoculated in and carry out inducing clumping bud and shoot proliferation on the shoot proliferation medium; Per 40～50 days is a successive transfer culture cycle, and the shoot proliferation medium is meant MS+ZT 0.5～1.5mg/L+IBA0.05～0.1mg/L+ sucrose 30g/L+ agar 6.5g/L, and regulating its pH is 5.2～5.8; 2～3 successive transfer culture adopt MS+ZT 0.3～0.7mg/L medium to carry out the nursery stock rejuvenation in the cycle before the blueberry bottle outlet is transplanted.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103548697A (en) * | 2013-11-07 | 2014-02-05 | 湖北省农业科学院果树茶叶研究所 | Culture medium for blueberry tissue culture |
| CN103548696A (en) * | 2013-11-05 | 2014-02-05 | 四川省农业科学院园艺研究所 | Method for quickly and efficiently breeding blueberry seedlings |
| CN103598093A (en) * | 2013-10-31 | 2014-02-26 | 李本明 | Inducing method of blueberry embryoid |
| CN103875529A (en) * | 2014-02-18 | 2014-06-25 | 杭州佑国生物科技有限公司 | Blueberry tissue culture propagation and ex-vitro rooting method |
| CN104012411A (en) * | 2014-06-09 | 2014-09-03 | 赵兰 | Tissue culture method for cowberry blueberries |
| CN105075865A (en) * | 2015-09-22 | 2015-11-25 | 江苏农林职业技术学院 | Blomidon blueberry primary culture medium and preparation method and application thereof |
| CN106171980A (en) * | 2016-07-12 | 2016-12-07 | 安徽师范大学 | Blueberry tissue cultural method |
| CN106258996A (en) * | 2016-10-21 | 2017-01-04 | 北京农业职业学院 | Blue berry stem section method for quickly breeding |
| CN107347639A (en) * | 2017-06-27 | 2017-11-17 | 丹东天赐花卉有限公司 | A kind of poison-removing method and cultural method of blueberry tissue culture seedling |
| CN115005105A (en) * | 2022-07-26 | 2022-09-06 | 辽宁省果树科学研究所 | Blueberry tissue culture method |
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| CN101869078A (en) * | 2010-07-14 | 2010-10-27 | 长春百瑞蓝莓科技发展有限公司 | Seed-cultivating method of blueberry by means of tissue cultivation and micropropagation |
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Patent Citations (1)
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Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103598093A (en) * | 2013-10-31 | 2014-02-26 | 李本明 | Inducing method of blueberry embryoid |
| CN103598093B (en) * | 2013-10-31 | 2015-10-21 | 青岛文创科技有限公司 | A kind of abductive approach of blueberry embryoid |
| CN103548696A (en) * | 2013-11-05 | 2014-02-05 | 四川省农业科学院园艺研究所 | Method for quickly and efficiently breeding blueberry seedlings |
| CN103548696B (en) * | 2013-11-05 | 2015-12-02 | 四川省农业科学院园艺研究所 | A kind of method of breeding blueberry seedling |
| CN103548697A (en) * | 2013-11-07 | 2014-02-05 | 湖北省农业科学院果树茶叶研究所 | Culture medium for blueberry tissue culture |
| CN103875529A (en) * | 2014-02-18 | 2014-06-25 | 杭州佑国生物科技有限公司 | Blueberry tissue culture propagation and ex-vitro rooting method |
| CN104012411B (en) * | 2014-06-09 | 2016-06-08 | 赵兰 | The tissue culture method of a kind of cowberry |
| CN104012411A (en) * | 2014-06-09 | 2014-09-03 | 赵兰 | Tissue culture method for cowberry blueberries |
| CN105075865A (en) * | 2015-09-22 | 2015-11-25 | 江苏农林职业技术学院 | Blomidon blueberry primary culture medium and preparation method and application thereof |
| CN106171980A (en) * | 2016-07-12 | 2016-12-07 | 安徽师范大学 | Blueberry tissue cultural method |
| CN106258996A (en) * | 2016-10-21 | 2017-01-04 | 北京农业职业学院 | Blue berry stem section method for quickly breeding |
| CN106258996B (en) * | 2016-10-21 | 2018-06-29 | 北京农业职业学院 | Blueberry stem section rapid propagation method |
| CN107347639A (en) * | 2017-06-27 | 2017-11-17 | 丹东天赐花卉有限公司 | A kind of poison-removing method and cultural method of blueberry tissue culture seedling |
| CN107347639B (en) * | 2017-06-27 | 2019-08-13 | 丹东天赐花卉有限公司 | A kind of poison-removing method and cultural method of blueberry tissue culture seedling |
| CN115005105A (en) * | 2022-07-26 | 2022-09-06 | 辽宁省果树科学研究所 | Blueberry tissue culture method |
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