CN101869078B - Seed-cultivating method of blueberry by means of tissue cultivation and micropropagation - Google Patents
Seed-cultivating method of blueberry by means of tissue cultivation and micropropagation Download PDFInfo
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- CN101869078B CN101869078B CN2010102293460A CN201010229346A CN101869078B CN 101869078 B CN101869078 B CN 101869078B CN 2010102293460 A CN2010102293460 A CN 2010102293460A CN 201010229346 A CN201010229346 A CN 201010229346A CN 101869078 B CN101869078 B CN 101869078B
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Abstract
The invention provides a seed-cultivating method of the blueberry by means of tissue cultivation and micropropagation, which comprises the following steps of: picking young sprout of the blueberry from fields in the growth season; sterilizing the surface of the young sprout under the aseptic condition; washing the sterilized young sprout with the sterile water; cutting the washed young sprout into a stem section with 1-2 bud(s); characteristically inoculating the stem section with 1-2 bud(s) on a first-generation inducting culture medium 3.0mg/L of WPM+ZT+0.02mg/L of NAA+30g/L of saccharose+6.5g/L of agar, wherein the PH value of the culture medium is 5.4; cultivating for 2-3 generations to obtain a tissue cultivation clone which is sterile and exuberantly grown and has multiple branches; using a cultivating culture medium 0.5-1.5mg/L of WPM+ZT+0.05-1.0mg/L of CPPU+30g/L of saccharose+6.5g/L of agar at the subculture and multiplication culture stage of the tissue cultivating seed of the blueberry, wherein the PH value of the culture medium is 5.4; and subculturing at a time every 50-60 days, thereby improving the propagation line by 5-10 times.
Description
Technical field
The present invention relates to the blueberry propagation method, what be specifically related to is seed-cultivating method of blueberry by means of tissue cultivation and micropropagation.
Background technology
Blueberry belongs to Ericaceae 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, speed is slow, can't satisfy market to the growing needs of blueberry nursery stock quantity.Utilizing tissue culture technique to breed the blueberry nursery stock fast is the most effective approach.The seedling quality of the little numerous production of group training is good, and uniformity is sloughed virus easily, and well developed root system will be set the bulk-growth result and lay a good foundation for the later stage.
As everyone knows, plant cell has totipotency, under stripped and manual control condition, can carry out continuous vegetative propagation.Under aseptic condition blueberry young sprout stem apex or stem section be inoculated on the medium of plant hormone such as the additional zeatin of WPM, condition is waited until in Artificial Control temperature, illumination, makes its proliferate constantly, can reach the purpose of a large amount of production nursery stocks.Normally used propagation hormone and growth regulator have: zeatin (ZT), 6-benzyl aminopurine (6-BA), 2-isopentennyladenine (2ip) etc., their specific activity is lower, the different factors of reproduction speed lower (being generally about the 5 times) Wang Daping of blueberry were induced the influence [J] of effect when tissue culture propagation was used to lagophthalmos blueberry cultured in vitro bud. Anhui agricultural science, 200937 (15); Blueberry kinds such as Duan Zuan ' the reach research [J] of gentle ' cultured in vitro fast breeding technique system. Shandong Agricultural University's journal (natural science edition), 200839 (4).
Summary of the invention
The objective of the invention is to solve the problem that prior art exists, provide a kind of blueberry group training little method of breeding seedling, improve seedling propagation speed.
Technical scheme of the present invention is: a kind of seed-cultivating method of blueberry by means of tissue cultivation and micropropagation, the season of growth is gathered the blueberry young sprout from the field, under the aseptic condition, through surface sterilization, after dashing with sterile water, cut into stem section, it is characterized in that and to be with 1～2 leaf stem section to be inoculated into just on the inducing culture WPM+ZT 3.0mg/L+NAA 0.02mg/L+ sucrose 30g/L+ agar 6.5g/L this medium pH 5.4 with 1～2 bud; Through the cultivation in 2～3 generations, obtain the group training clone aseptic, vigorous growth, that branch is many; At blueberry tissue cultivating seedling shoot proliferation cultivation stage, adopt proliferated culture medium WPM+ZT 0.5～1.5mg/L+CPPU 0.05～1.0mg/L+ sucrose 30g/L+ agar 6.5g/L, pH5.4; Per 50～60 days subculture cycles.
The present invention compared with prior art has outstanding substantive distinguishing features and marked improvement and good effect:
The present invention is little when numerous in group training, and with the CPPU plant growth regulator, its specific activity 6-benzyl aminopurine is high 10～100 times, has the promotion cell division, promotes cell to enlarge effects such as elongation.Evidence, CPPU can well induce the propagation and the regenerated adventitious bud of blueberry bud.The present invention adopts CPPU0.05～1.0mg/L to mix use with ZT0.5～1.5mg/L, and reproduction coefficient improves 5～10 times, has accelerated reproduction speed.
Embodiment
Embodiment
A kind of seed-cultivating method of blueberry by means of tissue cultivation and micropropagation of the present invention, the season of growth is gathered the blueberry young sprout from the field, under aseptic condition, through the surface sterilization in 5～7 minutes of mass percent 0.1% mercury chloride, after using aseptic water washing 3 times again, cut into and be with 1～2 leaf stem section to be inoculated into just for inducing culture WPM+ZT 3.0mg/L+NAA 0.02mg/L+ sucrose 30g/L+ agar 6.5g/L, on the pH 5.4.Through the cultivation in 2～3 generations, obtain the group training clone aseptic, vigorous growth, that branch is many.At blueberry tissue cultivating seedling shoot proliferation cultivation stage, adopt proliferated culture medium WPM+ZT0.5-1.5mg/L+CPPU 0.05-1.0mg/L+ sucrose 30g/L+ agar 6.5g/L, pH 5.4.Per 50～60 days subculture cycles (repeating identical incubation).2～3 successive transfer culture adopt WPM+ZT0.3～0.7mg/L medium to carry out the nursery stock rejuvenation in the cycle before the blueberry bottle outlet is transplanted.Culturing room's temperature is controlled at about 25 ℃, adopts the illumination of sight lamp, intensity of illumination 2000-4000Lux, illumination 16 hours every days, 8 hours dark.
Claims (1)
1. seed-cultivating method of blueberry by means of tissue cultivation and micropropagation, the season of growth is gathered the blueberry young sprout from the field, under the aseptic condition, through surface sterilization, after dashing with sterile water, cut into and be with 1～2 leaf stem section, it is characterized in that the stem section with 1～2 bud being inoculated into just on the inducing culture WPM+ZT3.0mg/L+NAA 0.02mg/L+ sucrose 30g/L+ agar 6.5g/L this medium pH 5.4; Through the cultivation in 2～3 generations, obtain the group training clone aseptic, vigorous growth, that branch is many; At blueberry tissue cultivating seedling shoot proliferation cultivation stage, adopt proliferated culture medium WPM+ZT 0.5～1.5mg/L+CPPU 0.05～1.0mg/L+ sucrose 30g/L+ agar 6.5g/L, pH 5.4; Per 50～60 days subculture cycles; 2～3 successive transfer culture adopt WPM+ZT0.3～0.7mg/L medium to carry out the nursery stock rejuvenation in the cycle before the blueberry bottle outlet is transplanted.
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| CN101869078B true CN101869078B (en) | 2011-12-21 |
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Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102138529B (en) * | 2011-03-31 | 2013-01-23 | 中国农业大学 | Quick rooting method for 'Heyun' blueberry seedlings formed by tissue culture |
| CN102187813B (en) * | 2011-03-31 | 2012-11-28 | 中国农业大学 | Blueberry tissue culture method and special culture medium thereof |
| CN102524063A (en) * | 2011-12-19 | 2012-07-04 | 普罗米绿色能源(深圳)有限公司 | Method for effectively inducing regeneration of adventitious bud of eucalyptus stem segment in vitro |
| CN102524077A (en) * | 2012-02-29 | 2012-07-04 | 东北林业大学 | Methods for quickly rooting and exercising tissue culture seedlings of lowbush blueberries |
| CN102823495A (en) * | 2012-09-10 | 2012-12-19 | 麻江县黔甬蓝莓有限公司 | Culture method for blueberries |
| CN102812905A (en) * | 2012-09-14 | 2012-12-12 | 四川立益生态农业开发有限公司 | Blueberry tender stem tissue culturing and rapid propagation process |
| CN103583368B (en) * | 2013-11-22 | 2016-03-30 | 西南林业大学 | The construction method of vaccinium ashei tissue culture rapid propagation system |
| CN104521542A (en) * | 2015-01-21 | 2015-04-22 | 玉林师范学院 | In vitro fast cultivation method for immature stem segments of blueberries |
| CN104585039B (en) * | 2015-02-09 | 2017-05-03 | 上海青浦现代农业园区发展有限公司 | Tissue culture and rapid propagation method of blueberry |
| CN106171980A (en) * | 2016-07-12 | 2016-12-07 | 安徽师范大学 | Blueberry tissue cultural method |
| CN106258996B (en) * | 2016-10-21 | 2018-06-29 | 北京农业职业学院 | Blueberry stem section rapid propagation method |
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| JP2003304765A (en) * | 2002-04-17 | 2003-10-28 | Gumma Prefecture | Method for raising seedling of blueberry |
| CN101069487A (en) * | 2007-06-20 | 2007-11-14 | 金陵科技学院 | Labbiteye blueberry tissue culturation method |
| CN101176428B (en) * | 2007-11-28 | 2011-08-24 | 浙江省农业科学院 | Culture medium composition adapting for blueberry test tube plantlet proliferation as well as method thereof |
| CN101305693B (en) * | 2008-07-08 | 2010-11-03 | 山东省果树研究所 | High irrigation blueberry cultured shoots one-step seedling establishment method |
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