CN102090337A - Rapid propagation method of rhododendron latoucheae - Google Patents
Rapid propagation method of rhododendron latoucheae Download PDFInfo
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- CN102090337A CN102090337A CN 201010584915 CN201010584915A CN102090337A CN 102090337 A CN102090337 A CN 102090337A CN 201010584915 CN201010584915 CN 201010584915 CN 201010584915 A CN201010584915 A CN 201010584915A CN 102090337 A CN102090337 A CN 102090337A
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Abstract
The invention relates to a tissue culture and rapid propagation method of rhododendron latoucheae. A woody plant medium (WPM) is taken as a basic culture medium, and a tissue culture and rapid propagation system of the rhododendron latoucheae is established through the isolated culture of the rhododendron latoucheae by proportioning regulation of different hormones. The method sequentially comprises the following steps of: selecting an explant; disinfecting; performing primary culturing; performing propagation culturing; performing rooting cultivation on a strong seedling; and transplanting to obtain a test tube seedling which can keep the original plant genotype, has a high aesthetic property and is better differentiated, wherein a primary induction culture medium of the explant is WPM+IBA 0.15mg/L+TDZ 0.3mg/L+cane sugar 30g/L and the pH value is 5.6; a propagation culture medium of an adventitious bud is WPM+IBA 0.15mg/L+TDZ 0.3mg/L+cane sugar 30g/L and the pH value is 5.6; and a rooting culture medium of the strong seedling is 1/2WPM+IBA 0.5mg/L+NAA 0.2mg/L+cane sugar 20g/L+activated carbon 0.3g/L and the pH value is 5.6. By the method, the propagation speed of the rhododendron latoucheae is greatly increased; and the method is suitable for industrialized commercial production of the rhododendron latoucheae.
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)
1. the quick-breeding method of a deer horn cuckoo is characterized in that by the cultured in vitro to the deer horn cuckoo, utilizes different hormone combination regulation and control, sets up the tissue-culturing quick-propagation system of deer horn cuckoo, and described method comprises following 4 cultivation stages:
(1) the clean deer horn cuckoo explant of cancellation poison is removed 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, oblique cutting in explant just for inducing culture (WPM+IBA 0.15mg/L+TDZ 0.3mg/L+ sucrose 30g/L) on, 1～2 explant of every bottle graft kind;
(2) indefinite bud that induces is changed over to carry out enrichment culture in the adventitious buds proliferation medium, it is identical that adventitious buds proliferation medium and explant start the medium of cultivating, and is WPM+IBA 0.15mg/L+TDZ 0.3mg/L+ sucrose 30g/L; The base portion of indefinite bud grows callus earlier, differentiates the seedling of growing thickly from callus, and 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, and each bud can be bred more than 5 times, so repeatedly, can obtain a large amount of seedlings of growing thickly;
(3) bud of growing thickly of propagation back robust growth is cut into individual plant, change root media (1/2WPM+IBA 0.5mg/L+NAA 0.2mg/L+ sucrose 20g/L+ active carbon 0.3g/L) over to and carry out culture of rootage, cultivate the adventive root that grows varying length about 40d, rooting rate is 85.4%;
(4) has the seedling of taking root of 4 above roots, move into the greenhouse, uncork goes out transplantation of seedlings behind the hardening of becoming less severe under normal temperature, the natural daylight 1～2d, and 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, matrix is disinfected with 0.1% carbendazim in advance, water permeablely, cover plastic film for agricultural use and preserve moisture, illumination is crossed and is suitably shaded when strong, open film gradually after cultivating about 1 week, ventilated.
2. deer horn cuckoo quick breeding method for tissue culture according to claim 1, it is characterized in that the deer horn cuckoo spray of getting robust growth, no damage by disease and insect does explant, dip in liquid detergent with banister brush and carefully scrub, especially the axil place uses running water drip washing 10～12h again.
3. deer horn cuckoo quick breeding method for tissue culture according to claim 1 is characterized in that on the desinfection chamber super-clean bench, and with the explant 30s of 70% alcohol processing drip washing, aseptic water washing once; Use 0.1% HgCl again
2(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.
4. deer horn cuckoo quick breeding method for tissue culture according to claim 1, the medium that it is characterized in that initial culture, enrichment culture, culture of rootage all adds agar 5g/L, pH 5.6, each triangular flask packing 30mL, in 121 ℃ of autoclave sterilization 20min, culturing room's temperature is (24 ± 1) ℃, light application time 12h/d, and intensity of illumination is 2000LX.
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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 rhododendron dauricum adventitious bud inducing 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 |
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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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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 rhododendron dauricum adventitious bud inducing and plant regeneration |
| CN109618932B (en) * | 2019-01-11 | 2021-06-18 | 东北林业大学 | Method for inducing adventitious buds of rhododendron dauricum and regenerating plants |
| 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 |