CN1135917C - Tissue culture method for breeding Dengen eucalyptus - Google Patents
Tissue culture method for breeding Dengen eucalyptus Download PDFInfo
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- CN1135917C CN1135917C CNB021141428A CN02114142A CN1135917C CN 1135917 C CN1135917 C CN 1135917C CN B021141428 A CNB021141428 A CN B021141428A CN 02114142 A CN02114142 A CN 02114142A CN 1135917 C CN1135917 C CN 1135917C
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Abstract
The present invention relates to a tissue culture method for breeding Dengen eucalyptus groups, which belongs to a method of plant tissue culture. In the method, an excellent single plant is chosen to be used as materials, and axillary buds of the single plant are chosen to be used as explants; the explants are washed by CaCl2 after being disinfected by HgCl2; in primary culture, subculture and breeding culture, MS+BA+IBA is used as a culture medium, and MS+IBA+NAA+active carbon is used as a root medium; the illumination intensity of 1000Lux, 2500Lux and 1500Lux are respectively used for the primary culture, the subculture and root culture; a test-tube plantlet is transplanted to a mixed base material of yellow subsoil and bran shell dust to be bred after being bred for about 25 days in the root media. In the present invention, the problem of low propagation coefficients of test-tube plantlets is solved, and the scale production of excellent Dengen seedlings is realized.
Description
Technical field: tissue culture method for breeding Dengen eucalyptus belongs to method for plant tissue culture, specifically is the method for tissue culture of a kind of eucalyptus.
Background technology: Dunne eucalyptus (Eucalyptus dunnii) is that a kind of speed is given birth to the reproducting tree species that material is good.But because Dunne eucalyptus is a cross pollinated species, use seminal propagation, individual differentiation is big, therefore in fast-growing andhigh-yeilding trees is built, easily causes speed to give birth to and does not get bumper crops; It is solid poor to add Dunne eucalyptus, and seed production is low, and seed costs an arm and a leg, and has more limited applying of these seeds.
Adopt conventional cottage propagation, survival rate is lower, below 5%, does not have using value aborning.
Adopting tissue culture method is the main path of exploring Dunne eucalyptus fast asexual propagation; though both at home and abroad the group training research of Dunne eucalyptus kind is being carried out always; but because tissue culture technology is not broken through, the reproduction coefficient of test-tube plantlet is low, thereby has hindered the carrying out of Eucalypts camaldulensis,E. seedlings large-scale production.
Summary of the invention: the present invention is intended to explore a kind of tissue culture method, makes the moon growth coefficient of Dunne's cineole pipe seedling and propagating reach requirement, thereby has solved the large-scale production problem of Eucalypts camaldulensis,E. seedlings.
The foregoing invention purpose that the present invention proposes realizes by following scheme.
Tissue culture method for breeding Dengen eucalyptus comprises explant sterilization, cultivation just, successive transfer culture, culture of rootage and test-tube seedling transplanting, is explant with the axillalry bud, through HgCl
2After the sterilization, under aseptic condition with 0.25～1% CaCl
2Clean several times, just cultivating is to carry out on MS+BA0.1～0.3mg/l+IBA0.8～1.2mg/l medium, intensity of illumination is 1000 ± 200Lux, the medium of successive transfer culture and expanding propagation also is MS+BA0.1～0.3mg/l+IBA0.8～1.2mg/l, intensity of illumination is 2500 ± 200Lux, the medium in culture of rootage stage is MS+IBA0.1～0.2mg/l+NAA0.1～0.2mg/l+ active carbon 2～4g/l, intensity of illumination is 1500 ± 200Lux, test-tube plantlet is cultivated on root media, be transplanted to when treating root growth to 0.3～0.5cm on the matrix of yellow soil bran adding shell ash and cultivate, the volume ratio of yellow soil and chaff shell ash is 2: 1.
Be described in further detail the present invention below.
Dunne eucalyptus group training material is to select the axillalry bud of Dunne eucalyptus fine individual plant to do explant, when carrying out the sterilization of explant, after 20 minutes, uses 0.25% HgCl with the clear water cleaning more earlier
2Sterilized 15 minutes, then under aseptic condition with 0.25～1% CaCl
2Clean 3 times, to displace the mercury ion that remains on the explant, use sterile water wash again 3 times, explant through sterilization, under aseptic condition, be inoculated on the medium of MS+BA0.1～0.3mg/l+IBA0.8～1.2mg/l, at the beginning of under the illumination condition of 1000 ± 200Lux, cultivate, approximately growth is about one month, axillalry bud up to 2cm about, can enter successive transfer culture, expand numerous test-tube plantlet, the medium that successive transfer culture adopted is with just cultivating, also be MS+BA0.1～0.3mg/l+IBA0.8～1.2mg/l, but it is 2500 ± 200Lux that intensity of illumination strengthens, under such condition of culture, the moon growth coefficient of test-tube plantlet is about 3, therefore only needed 1 year just can breed test-tube plantlet to 500,000 strains from an explant, for condition has been created in the large-scale production of Eucalypts camaldulensis,E. seedlings, waiting to expand numerous test-tube plantlet breeds after some, the test-tube plantlet of choosing certain altitude carries out culture of rootage, root media is MS+IBA0.1～0.2mg/l+NAA0.1～0.2mg/l+ active carbon 2～4g/l, test-tube plantlet is in root media, began to take root in about 15 days cultivating under 1500 ± 200Lux illumination condition, carry out test-tube seedling transplanting when treating root growth to 0.3～0.5cm to about 25 days: at the test-tube plantlet cultivation stage, cultivation temperature is 25 ± 3 ℃, medium PH is 5.8～6.0, and light application time is about 14 hours.Cultivate on the grey matrix of mixing of yellow soil and chaff shell at cultured test-tube seedling transplanting on the root media, yellow soil is taken from the soil of slaking under the plow layer, the volume ratio of yellow soil and chaff shell ash is 2: 1, such matrix, not only ventilative water permeability is good, and the essential phosphorus of seedling growth, potassium and other nutritive elements are arranged, also can reduce the harmful microbe invasion and attack; Test-tube seedling transplanting on the matrix after, irrigate with clear water, every medicine of spray in 3 days, prevent that damage by disease and insect from taking place, spray medicine and be penicillin, fenaminosulf, carbendazim and use in turn for three kinds; Transplanting the place is the cement flooring, should be stamped plastic tunnel, fine sun-proof, the insulation in winter.The temperature of transplanting breeding is controlled at 0～30 ℃, crosses to exceed low the surviving of test-tube plantlet that be unfavorable for.Under the temperature suitable condition, to transplant after 20 days, test-tube plantlet promptly begins growth, and later management is identical with general nursery stock management.
Cultivate Dunne eucalyptus test-tube plantlet with tissue culture method of the present invention; solved the low difficult problem of reproduction coefficient; the moon growth coefficient of test-tube plantlet is reached about 3; the group of Dunne eucalyptus training no longer is the laboratory research achievement like this; can satisfy the requirement of Eucalypts camaldulensis,E. seedlings large-scale production fully; for the extensive use of tissue culture method large-scale production Eucalypts camaldulensis,E. seedlings has proposed practicable technical scheme; therefore; the present invention has realized the production scaleization of the high-quality Eucalypts camaldulensis,E. seedlings of fast life, resistance, has alleviated the anxiety of current nurseries.
Embodiment:
Organize the preferred forms of cultivating seedling for Dunne eucalyptus of the present invention below.
The axillalry bud of Dunne eucalyptus fine individual plant of choosing, through flushing with clean water, 0.25%HgCl
2After the sterilization, use 1%CaCl
2Under aseptic condition, clean 3 times, use sterile water wash again, the first cultivation of explant and successive transfer culture be the medium of available MS+BA0.2mg/l+IBA1.0mg/l all, root media is MS+IBA0.15mg/l+NAA0.15mg/l+ active carbon 3g/l, test-tube plantlet is cultivated on root media, transplant to yellow soil when treating root growth to 0.3～0.5cm: cultivate on the matrix of chaff shell ash=2: 1 (volume ratio), under the temperature suitable condition, transplant and to grow in back 20 days.
Claims (1)
1, a kind of tissue culture method for breeding Dengen eucalyptus comprises explant sterilization, cultivation just, successive transfer culture, culture of rootage and test-tube seedling transplanting, it is characterized in that:
(1) the explant axillalry bud is through HgCl
2After the sterilization, under aseptic condition with 0.25～1% CaCl
2Clean several times,
(2) just cultivating is to carry out on MS+BA0.1～0.3mg/l+IBA0.8～1.2mg/l medium, and intensity of illumination is 1000 ± 200Lux,
(3) medium of successive transfer culture and expanding propagation is MS+BA0.1～0.3mg/l+IBA0.8～1.2mg/l, and intensity of illumination is 2500 ± 200Lux,
(4) medium of culture of rootage is MS+IBA0.1～0.2mg/l+NAA0.1～0.2mg/l+ active carbon 2～4g/l, and intensity of illumination is 1500 ± 200Lux,
(5) test-tube plantlet is cultivated on root media, transplants to matrix when treating root growth to 0.3～0.5cm and cultivates, and transplanting medium is a yellow soil bran adding shell ash, and the two volume ratio is 2: 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CNB021141428A CN1135917C (en) | 2002-05-22 | 2002-05-22 | Tissue culture method for breeding Dengen eucalyptus |
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| CNB021141428A CN1135917C (en) | 2002-05-22 | 2002-05-22 | Tissue culture method for breeding Dengen eucalyptus |
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| CN1381171A CN1381171A (en) | 2002-11-27 |
| CN1135917C true CN1135917C (en) | 2004-01-28 |
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Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101658137B (en) * | 2009-08-31 | 2011-11-23 | 重庆文理学院 | Flooded gum clone WLEG-17 tissue culture medium |
| CN102057871B (en) * | 2010-11-08 | 2013-01-23 | 澧县万红桉树林业科技发展有限公司 | Method for breeding cold-resistant eucalypt Xiang eucalypt No.1 variety suitable for north latitude 26 to 30 degrees |
| CN102388738B (en) * | 2011-07-29 | 2013-04-10 | 广州长隆集团有限公司香江野生动物世界分公司 | Eucalyptus cultivation method |
| CN104082299B (en) * | 2014-07-09 | 2016-05-18 | 湖南省森林植物园 | A kind of method of plant growth regulator for E. dunnii cuttage and seedling culture and E. dunnii cuttage and seedling culture |
| CN105638475B (en) * | 2016-01-22 | 2018-05-18 | 中国农业科学院生物技术研究所 | The wide 29 stem apex detoxification of woods of Eucalyptus urophylla-grandis and quick proliferation method |
| CN105724252B (en) * | 2016-03-02 | 2018-01-05 | 广西壮族自治区林业科学研究院 | One kind promotes E. dunnii tissue-cultured seedling rooting method |
| CN106614726A (en) * | 2016-10-12 | 2017-05-10 | 许伟琦 | Growth regulator of eucalyptus dunnii cutting seedling and method of eucalyptus dunnii cutting seedling |
| CN107660433A (en) * | 2017-09-27 | 2018-02-06 | 广东省林业科学研究院 | A kind of Eucalyptus urophylla seedling stage breeding method |
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