CN102090337A - Rapid propagation method of rhododendron latoucheae - Google Patents
Rapid propagation method of rhododendron latoucheae Download PDFInfo
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
本发明涉及一种鹿角杜鹃的组织培养快速繁殖方法。以WPM为基本培养基，通过对鹿角杜鹃的离体培养，利用不同激素配比调控，建立起鹿角杜鹃的组织培养快速繁殖体系。具体步骤依次为外植体的选择、消毒、初代培养、增殖培养、壮苗生根培养和移栽。获得能保持原植株基因型、优良观赏特性，且分化良好的试管苗。其中，外植体初代诱导培养基为WPM+IBA 0.15mg/L+TDZ 0.3mg/L+蔗糖30g/L，pH 5.6；不定芽增殖培养基为WPM+IBA 0.15mg/L+TDZ 0.3mg/L+蔗糖30g/L，pH 5.6；壮苗生根培养培养基为1/2WPM+IBA 0.5mg/L+NAA 0.2mg/L+蔗糖20g/L+活性炭0.3g/L，pH 5.6。本发明大大提高了鹿角杜鹃繁殖速度，适用于鹿角杜鹃的工厂化商业生产。The invention relates to a rapid propagation method of rhododendron staghorn through tissue culture. Using WPM as the basic medium, through the in vitro culture of Rhododendron antlers and the regulation of different hormone ratios, a tissue culture rapid propagation system of Rhododendron antlers was established. The specific steps are sequentially the selection of explants, disinfection, primary culture, proliferation culture, rooting culture of strong seedlings and transplanting. The test-tube plantlets that can maintain the genotype of the original plant, have excellent ornamental characteristics, and are well differentiated are obtained. Among them, the primary induction medium for explants is WPM+IBA 0.15mg/L+TDZ 0.3mg/L+sucrose 30g/L, pH 5.6; the adventitious bud proliferation medium is WPM+IBA 0.15mg/L+TDZ 0.3mg/L+ Sucrose 30g/L, pH 5.6; strong seedling rooting culture medium is 1/2WPM+IBA 0.5mg/L+NAA 0.2mg/L+sucrose 20g/L+activated carbon 0.3g/L, pH 5.6. The invention greatly improves the propagation speed of Rhododendron staghorn, and is suitable for industrialized commercial production of Rhododendron staghorn.
Description
Technical field
The present invention relates to the quick breeding method for tissue culture of a kind of deer horn cuckoo, belong to biological technical field.
Background technology
Rhododendron is the genus of Ericaceae maximum, contains about 960 kinds of species, is distributed widely in Europe, Asia, North America, main product East Asia and Southeast Asia.China is the modern center of differentiation and the country of population maximum of distributing of world's wild-type azalea germ plasm resource, about 542 kinds, and main product southwest, South China, pattern is gorgeous colourful, is famous ornamental plants, and the deer horn cuckoo is exactly one of them.
Deer horn cuckoo (Rhododendron latoucheae), another name rock cuckoo, because of its limb complications, shape such as deer horn, and gain the name.Evergreen shrubs or dungarunga, happiness mild climate, happiness acid soil; Bloom 3～April, large flower and brilliant color, and the florescence is longer, and is savory.Nature vertical distribution scope is wide, all can grow from prickly pine sylvan life, the shrubbery of 100～1200m, compares with other cuculiform other kind, and the deer horn cuckoo is one of the wideest kind of vertical distribution during cuckoo belongs to.Along with flowers and trees market is flourish, directly transplant when having part to excavate, transplant wild deer horn cuckoo from the mountain, lack scale, systemic propagation technique, transplanting survival rate is low, can not meet the need of market.
At present domestic about the breeding of deer horn cuckoo and the research of propagation technique aspect, the report of minority cutting propagation is only arranged, still there is not the report of deer horn cuckoo tissue culture technique research.The present invention is the batch production commodity production of deer horn cuckoo and promotes and prepare that making wild deer horn cuckoo is that afforestation is used, increases the diversity of landscape plant kind on a large scale, promotes landscape ecology municipal water equality aspect and all has crucial meaning.
Summary of the invention
Technical problem: the present invention is directed to deer horn cuckoo normal cutting propagation, the propagation by grafiting coefficient is low and complex operation is difficult present situation; provide a kind of reproduction speed fast; be not subjected to the deer horn cuckoo quick breeding method for tissue culture of seasonal effect; technical problem such as overcome in the group training process that sterile system sets up that difficulty is big, growth coefficient is low, the seedling of growing thickly growth is thin and delicate, the difficulty of taking root, transplanting survival rate are low is found out quick breeding method for tissue culture and the optimum medium of scale, low-cost production deer horn cuckoo.
Technical scheme: it is minimal medium that the present invention adopts WPM, by cultured in vitro to the deer horn cuckoo, set up the tissue-culturing quick-propagation system of deer horn cuckoo, concrete steps are followed successively by selection, sterilization, initial culture, enrichment culture, the strengthening seedling and rooting of explant and cultivate and acclimatization and transplants.The deer horn cuckoo is cooked explant and starts cultivation, the HgCl with 0.1%
2Sterilization on the basis of the sterile system that success is set up, is adjusted culture medium prescription the evoking adventive bud survival rate is increased to 100%; The adventitious bud proliferation of sprouting is cultivated, and grows the bud of growing thickly from callus, and shoot proliferation can obtain a large amount of seedlings of growing thickly so repeatedly; The seedling individual plant of will growing thickly is transferred on the strengthening seedling and rooting medium, turns out complete regeneration plant; Transplantation of seedlings will take root at last to by perlite and turfy soil in proportion in the matrix prepared.
The medium of deer horn cuckoo tissue-culturing quick-propagation mainly is following several medium:
1, explant is just for inducing culture
WPM+IBA 0.15mg/L+TDZ 0.3mg/L+ sucrose 30g/L+ agar 5g/L, pH 5.6
2, indefinite bud shoot proliferation medium
WPM+IBA 0.15mg/L+TDZ 0.3mg/L+ sucrose 30g/L+ agar 5g/L, pH 5.6
3, strengthening seedling and rooting culture medium
1/2WPM+IBA 0.5mg/L+NAA 0.2mg/L+ sucrose 20g/L+ active carbon 0.3g/L+ agar 5g/L, pH 5.6
Beneficial effect: utilize the method for tissue culture, adopt the adventitious buds proliferation method of the mitogenetic approach of indefinite bud, provide a kind of deer horn cuckoo to breed the effective way of scale breeding fast.This propagation method stabilization characteristics of genetics, can keep former plant genotype and the good characteristic of viewing and admiring preferably, and select the optimal medium hormone combination for use, make and test good reproducibility, the test-tube plantlet well differentiated has improved deer horn cuckoo reproduction speed and transplanting survival rate greatly.
Embodiment
The present invention adopts tender stem apex and stem section to form the method for quickly breeding of the bud of growing thickly, and further describes the present invention below in conjunction with example.
1, the preparation of explant
Material is taken from the deer horn cuckoo spray of robust growth, no damage by disease and insect in the cuckoo plantation greenhouse, resource garden, academy of agricultural sciences, Jiangsu Province and is done explant.Explant dips in liquid detergent with banister brush and carefully scrubs, and especially the axil place uses running water drip washing 10～12h again, and is standby.
2, the sterilization of explant
On the desinfection chamber super-clean bench, handle 30s with 70% alcohol, aseptic water washing is once; 0.1% HgCl2 (mercury chloride) soaks stem apex 5min, stem section 8min, during ceaselessly shake bottle, explant is fully contacted with bactericidal liquid, aseptic water washing is 5～6 times afterwards.Remove the part of blackening on stem apex and the stem section, put into the sterile petri dish that filter paper is housed and blot surface moisture, standby.
3, the startup of indefinite bud is cultivated
The explant that sterilization is good, oblique cutting in explant just for inducing culture (WPM+IBA 0.15mg/L+TDZ 0.3mg/L+ sucrose 30g/L+ agar 5g/L, pH 5.6) on, 1～2 explant of every bottle graft kind.Culturing room's temperature is (24 ± 1) ℃, light application time 12h/d, and intensity of illumination is 2000LX.Note during this time observing, reject the material that pollutes at any time, in order to avoid cross-infection.
4, the enrichment culture of indefinite bud
The indefinite bud that induces changes over to and carries out enrichment culture in the adventitious buds proliferation medium, the adventitious buds proliferation medium is identical with the medium that explant starts cultivation, be WPM+IBA 0.15mg/L+TDZ 0.3mg/L+ sucrose 30g/L+ agar 5g/L, pH 5.6, and condition of culture is constant.The base portion of indefinite bud grows callus earlier, differentiate the seedling of growing thickly from callus, the Miao Tuanzai of will growing thickly for a short time is transferred to subculture propagation in the proliferated culture medium, about 10 of every bottle grafts, each bud can be bred more than 5 times, shoot proliferation can obtain a large amount of seedling groups of growing thickly so repeatedly, reaches the purpose of quick breeding.
5, strong sprout and culture of rootage
The bud of growing thickly of propagation back robust growth is cut into individual plant, changes in the root media (1/2WPM+IBA 0.5mg/L+NAA 0.2mg/L+ sucrose 20g/L+ active carbon 0.3g/L+ agar 5g/L, pH 5.6), and strong sprout and culture of rootage are carried out simultaneously, and condition of culture is constant.Cultivate the adventive root that grows varying length about 40d, rooting rate is 85.4%.
6, hardening and transplanting
The seedling of taking root that will have 4 above roots moves into the greenhouse, and uncork goes out transplantation of seedlings to the dish of cave behind the hardening of becoming less severe under normal temperature, the natural daylight 1～2d.Flush away is attached to the medium of root, and field planting is in 1: 1 by volume matrix prepared of perlite and turfy soil, and matrix is disinfected with 0.1% carbendazim in advance.Water permeablely, cover plastic film for agricultural use and preserve moisture, illumination is crossed and suitably shaded when strong, opens film gradually after cultivating about 1 week, and is ventilated.The tissue cultivating seedling transplanting survival rate is about 90%.
Claims (4)
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| CN 201010584915 CN102090337A (en) | 2010-12-13 | 2010-12-13 | Rapid propagation method of rhododendron latoucheae |
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| CN 201010584915 CN102090337A (en) | 2010-12-13 | 2010-12-13 | Rapid propagation method of rhododendron latoucheae |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102960250A (en) * | 2012-11-30 | 2013-03-13 | 通化师范学院 | Method for efficiently forming seedlings in vitro by utilizing stem segments of rhododendron plants in Changbai Mountains |
| CN103125384A (en) * | 2011-12-02 | 2013-06-05 | 刘家迅 | Tissue culture and fast propagation method of South China Sea azalea |
| CN103371103A (en) * | 2013-07-30 | 2013-10-30 | 杭州植物园 | Rapid propagation method for tissue culture of Rhododendron delavayi Franch |
| CN105475138A (en) * | 2015-12-18 | 2016-04-13 | 深圳职业技术学院 | Rapid propagation method of Rhododendron moulmainense Hook. f. |
| CN105766295A (en) * | 2016-03-11 | 2016-07-20 | 台州学院 | Artificial breeding method for rhododendron huadingense seedlings |
| CN105830917A (en) * | 2016-03-24 | 2016-08-10 | 江苏省农业科学院 | Method of regeneration plant from rhododendron molle leaves |
| CN109618932A (en) * | 2019-01-11 | 2019-04-16 | 东北林业大学 | A kind of method of Xing'an Rhododendron adventitious bud induction and plant regeneration |
| CN115250908A (en) * | 2022-04-06 | 2022-11-01 | 贵州民族大学人文科技学院 | Tissue culture propagation method of extremely endangered plant rhododendron lychee |
| CN117571935A (en) * | 2024-01-15 | 2024-02-20 | 海南大学三亚南繁研究院 | Application of TDZ solution in-vitro identification of bacterial leaf blight resistance of rice and identification method |
Citations (1)
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| CN1568670A (en) * | 2004-04-26 | 2005-01-26 | 上海市闵行区农业科学研究所 | Tissue culture and tachytelic propagating method for alpine rhododendron |
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2010
- 2010-12-13 CN CN 201010584915 patent/CN102090337A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1568670A (en) * | 2004-04-26 | 2005-01-26 | 上海市闵行区农业科学研究所 | Tissue culture and tachytelic propagating method for alpine rhododendron |
Non-Patent Citations (3)
| Title |
|---|
| 《亚热带植物科学》 20071231 吴连松等 二乔杜鹃无菌株系建立的初步研究 39-41 1-4 第36卷, 第2期 2 * |
| 《扬州大学学报（农业与生命科学版）》 20071231 刘晓青等 高山杜鹃茎段组织培养和优化体系的建立 91-94 1-4 第28卷, 第3期 2 * |
| 《湖南林业科技》 20091231 向光锋等 鹿角杜鹃的扦插试验 13-14 1-4 第36卷, 第6期 2 * |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103125384A (en) * | 2011-12-02 | 2013-06-05 | 刘家迅 | Tissue culture and fast propagation method of South China Sea azalea |
| CN102960250A (en) * | 2012-11-30 | 2013-03-13 | 通化师范学院 | Method for efficiently forming seedlings in vitro by utilizing stem segments of rhododendron plants in Changbai Mountains |
| CN102960250B (en) * | 2012-11-30 | 2014-02-26 | 通化师范学院 | Method for efficiently forming seedlings in vitro by utilizing stem segments of rhododendron plants in Changbai Mountains |
| CN103371103A (en) * | 2013-07-30 | 2013-10-30 | 杭州植物园 | Rapid propagation method for tissue culture of Rhododendron delavayi Franch |
| CN105475138A (en) * | 2015-12-18 | 2016-04-13 | 深圳职业技术学院 | Rapid propagation method of Rhododendron moulmainense Hook. f. |
| CN105766295A (en) * | 2016-03-11 | 2016-07-20 | 台州学院 | Artificial breeding method for rhododendron huadingense seedlings |
| CN105830917A (en) * | 2016-03-24 | 2016-08-10 | 江苏省农业科学院 | Method of regeneration plant from rhododendron molle leaves |
| CN105830917B (en) * | 2016-03-24 | 2017-11-17 | 江苏省农业科学院 | The method of Chinese azalea leaf regeneration plant |
| CN109618932A (en) * | 2019-01-11 | 2019-04-16 | 东北林业大学 | A kind of method of Xing'an Rhododendron adventitious bud induction and plant regeneration |
| CN109618932B (en) * | 2019-01-11 | 2021-06-18 | 东北林业大学 | A kind of method of Xing'an Rhododendron adventitious bud induction and plant regeneration |
| CN115250908A (en) * | 2022-04-06 | 2022-11-01 | 贵州民族大学人文科技学院 | Tissue culture propagation method of extremely endangered plant rhododendron lychee |
| CN115250908B (en) * | 2022-04-06 | 2023-09-08 | 贵州民族大学人文科技学院 | Tissue culture propagation method of extremely endangered plant azalea |
| CN117571935A (en) * | 2024-01-15 | 2024-02-20 | 海南大学三亚南繁研究院 | Application of TDZ solution in-vitro identification of bacterial leaf blight resistance of rice and identification method |
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Application publication date: 20110615 |