CN101530063A - Rapid propagation method for clonal tissue culture of oil tea - Google Patents
Rapid propagation method for clonal tissue culture of oil tea Download PDFInfo
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- CN101530063A CN101530063A CN200910043029A CN200910043029A CN101530063A CN 101530063 A CN101530063 A CN 101530063A CN 200910043029 A CN200910043029 A CN 200910043029A CN 200910043029 A CN200910043029 A CN 200910043029A CN 101530063 A CN101530063 A CN 101530063A
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Abstract
The invention relates to a rapid propagation method for clonal tissue culture of oil tea, comprising four stages of primary generation, secondary generation, rootage and transplantation; wherein the preparation of cutting rootage of tissue culture plantlets and rootage matrix comprises the following steps: the rootage matrix is prepared by yellow-core soil, carbonized rice husk ash and fine sand with the volume ratio of 1:2 to 1: 3; the matrix is put into a cutting container and the tissue culture plantlets are cut in the container and sealed by a transparent film. The rootage and transplantation of the tissue culture plantlets are integrated into a whole body; after 30 to 40 days, the tissue culture plantlets take roots and form complete plants, therefore, the section of transplantation of the tissue culture plantlets is saved and the rootage rate and survival rate are more than 87 percent.
Description
Technical field
The present invention relates to a kind of clonal tissue culture of oil tea fast breeding technique, specifically provide a kind of method that obtains a large amount of Camellia Oleifera Clones nursery stocks by tissue-culturing quick-propagation.
Background technology
Oil tea (Camellia spp) is seed oil content height in Theaceae (Theacae) Camellia (Camellia), have the general name of the oil of productive value with species, be the distinctive high-quality woody oil tree species of China, with oil palm, coconut and olive and be called the world's four big woody oil tree species
[1]4,531 ten thousand mu of the existing oil tea area under cultivations of China, mainly be distributed in south 14 provinces (district), the tea oil unique flavor that the oil tea seed squeezes, nutritious, fatty acid is formed rationally, unsaturated fatty acid content is up to 90%, oleic acid content does not contain erucic acid and mountain Yu's acid that human body is difficult to absorb more than 80%, does not contain to cause that human blood-pressure increases and causes angiosclerotic cholesterol yet, oil is better than other edible oil of olive wet goods, is desirable edible oil
[2]The by-product of tea oil is sampled tea withered and the tea shell can also extract Tea Saponin by comprehensive utilization, grind scrape powder and fermentation high protein feed etc., oil-tea camellia husks can be produced activated carbon and extract tanning material etc., so can increase substantially the oil tea value-added content of product by deep processing and comprehensive utilization.Oil tea wide adaptability, barren-resistant is one of low mountain, south, the good economic forest seeds in hills and hillock, does not strive ground with grain, cotton, but also tool is planted one year, characteristics of Shou Yiing for many years, high yield period reaches 80 years, occupies very consequence in China's south rural economy and ecological construction
[3], the development oil tea produces and helps alleviating the long-term dependence on import of China's edible oil, and the contradiction of supply and demand anxiety is significant to improving Chinese national economy and living standards of the people.
The oil tea seedling breeding mainly adopts seeding and seedling raising and sprout stock grafting.Past oil tea propagation method mainly is a seeding and seedling raising, and the offspring breaks up seriously, is difficult to keep the merit of maternal plant, and general 6～8 years beginnings, really output just differed.The oil tea seedling breeding mainly adopts vegetative propagation mode-sprout stock grafting at present, this method and technology difficulty is big, graft survival rate and qualified seedling outplanting rate are all lower, go out the garden for up to 2 years, survival rate instability, cost of forestation height, the forest form regularity is poor, the seedling propagation coefficient is low, is difficult to satisfy the demand of national camellia oleiferaindustry development to Camellia Oleifera Clones, presses for the efficient tissue-culturing rapid propagation new technology of research Camellia Oleifera Clones.
Group culturation rapid propagating technology is a kind of asexual reproduction method that is widely used in agricultural, forestry, has that reproduction speed is fast, method is simple, processing ease, and material consumption is few, is not subjected to the characteristics of season and region restriction, and filial generation simultaneously can keep the good hereditary capacity of maternal plant.Therefore, research Camellia Oleifera Clones group culturation rapid propagating technology to alleviating oil tea nursery stock anxiety, accelerating the camellia oleiferaindustry development, is alleviated the nervous situation of China's edible oil, promotes rural economy and new socialist countryside construction and has great strategic importance.Plant Tissue Breeding need be passed through first generation, subculture, be taken root, transplants four-stage, obtains whole plant.In the subculture stage, improve growth coefficient by adjusting minimal medium and plant hormone; Take root the stage, the general plant hormone that adds hestening rooting in medium was taken root about 30 days, and the stage of taking root is still aseptic, needed the preparation root media, expended a large amount of medicine expenses, charges for water and electricity, service charge etc.; Back test-tube plantlet is moved down from aseptic condition of taking root is planted in the outdoor environment, and its adaptive capacity is poor, and transplanting survival rate is low, causes the test-tube plantlet production cost than high several times or tens times of common seedling price.
Phase early 1980s, the domestic scholar of having has carried out research to the oil tea tissue culture, institute of woods section of Guangxi autonomous region face admire be diligent in 1980 the report, mainly induced embryoid by the oil tea tissue culture, cotyledon placed contain high concentration 2, cultivate in 4-D (mg/L) the SH medium, formed regeneration plant by embryoid
[4]1981 and nineteen eighty-two, grand shake male and Lu Tianling successively utilizes oil tea unmature subleaf rataria cultured in vitro respectively, adopt the additional NAA (0.05) of White+BA (0.5), the embryoid induction rate reaches 100%, employing N
6Additional IAA (0.5)+BA (2)+LH (500), differentiation rate reaches 100%
[5,6], after this, the grand hero that shakes is studied the oil tea anther culture, induces by tissue culture to have produced callus, and has obtained green bud point and root, but do not obtain regeneration plant
[7]The oil tea tissue culture is stagnated substantially subsequently, 2004, the Bi Fang tomahawk of Zhongnan Forestry Inst., Zhang Zhijun etc. induce under isolated condition stem section, rataria, the cotyledon of oil tea, stem section cultured in vitro is minimal medium with MS, additional 6-BA 3.0mg/L+NAA 1.0mg/L induces bud, and inductivity reaches 83.33%; Continuation subculture on this medium obtains the bud of growing thickly, and aseptic seedling is gone up root induction at MS medium (additional NAA 7mg/L), but the regrowth that obtains has only a main root mostly, fibrous root seldom, test-tube seedling transplanting is difficult to survive, so fail to obtain the plant of transplant survival
[8,9]Wu Zhengkai
[10]Find that Deng in the oil tea tissue culture procedures MV medium can root induction, but rooting rate only 72%, also fail to obtain the plant of transplant survival.
Show that according to prior art the tissue-culturing rapid propagation of Theaceae Camellia plant oil tea is difficulty comparatively, cell is difficult for inducing the differentiation regeneration plant, and the difficulty of taking root, and test-tube seedling transplanting is difficult to survive, and does not obtain the test-tube plantlet of transplant survival so far.The present invention is directed to the deficiencies in the prior art, provide a kind of simple to operate, reproduction coefficient height, rooting rate and survival rate height, the oil tea tissue-culturing rapid propagation new technology that the tissue cultivating seedling production cost is low.
Summary of the invention
The invention provides a kind of rapid propagation method for clonal tissue culture of oil tea, comprise just generation, subculture, take root, transplant four-stage, after material is taken root in acquisition, will take root and transplant step unites two into one, wherein:
The preparation of the tissue cultivating seedling cuttage root-taking and the matrix of taking root: be mixed with the matrix of taking root in 1: 2 by volume～3: 1 by yellow soil, charing rice husk ash, fine sand; Matrix is packed in the cuttage container, with the tissue cultivating seedling cuttage wherein, and seal with light transmission film.
Further preferred version is after tissue cultivating seedling is taken out from blake bottle, tissue cultivating seedling to be carried out disinfection.Preferably in 0.1% metalaxyl-MnZn solution, soaked 2-4 minute.
According to embodiments of the invention, also to the take root hormone preliminary treatment of material of tissue cultivating seedling.Preferably the base portion with tissue cultivating seedling soaked 3-4 seconds in the KIBA of 500～8000ppm solution.Further preferably, the matrix to the cuttage tissue cultivating seedling also carries out disinfection.
The invention also discloses the take root optimization of period management of tissue cultivating seedling.Place the green house of tool intermittent spraying facility to manage in the cuttage container of cuttage tissue cultivating seedling, temperature keeps 20～30 degree, after 30～40 days, and the tissue cultivating seedling formation whole plant of taking root.
Implement when of the present invention, tissue cultivating seedling used in the said method can obtain according to the method for announcing in the prior art.The present invention simultaneously also provides the method for preferred acquisition tissue cultivating seedling.
In the method for acquisition tissue cultivating seedling provided by the invention:
(1) subculture medium and hormone combination are that WPM+BA 3-7mg/L+IBA 0.1-2mg/L+ZT 0.05-2mg/L breeds;
(2) strong seedling culture base and hormone combination are WPM+IBA0.1-1mg/L+GA5-20mg/L; Root induction.
A kind of clonal tissue culture of oil tea propagation method provided by the invention can realize the tissue cultivating seedling formation whole plant of taking root, and saves the link that tissue cultivating seedling is transplanted, and rooting rate and survival rate reach more than 87%.
Description of drawings
Fig. 1-2 is the experimental result picture of oil tea tissue cultivating seedling vegetative state;
Fig. 3 is the experimental result picture of oil tea tissue cultivating seedling state in strong sprout;
Fig. 4 is the take root experimental result picture of shape of oil tea tissue cultivating seedling;
Fig. 5 is that the oil tea tissue cultivating seedling is taken root and survived the figure as a result of shape.
Embodiment
Embodiment 1
Clip is given birth to newly slightly then from the healthy and strong plant of oil tea, gets top 4-6cm disinfection; The aseptic seedling that first generation obtains is transferred to the WPM+BA3mg/L+IBA1.0mg/L+ZT0.5mg/L successive transfer culture, and be 30 days proliferating cycle, and growth coefficient reaches 4.6; Adopt WPM+IBA0.1mg/L+GA10mg/L to carry out strong sprout the aseptic seedling that obtains in the breeding, from blake bottle, take out when tissue cultivating seedling grew to 2-3cm in 20 days, its base portion is soaked 4s in the KIBA of 500ppm solution, with the nursery stock cuttage in yellow soil, charing rice husk ash, fine sand by volume in the matrix of 1:3:1, rooting rate 87% after 30 days.
Concrete operations are as follows:
(1) aseptic seedling that obtains is transferred on the subculture medium bred, medium is with the wide-mouth bottle packing of 300ml, and every bottle of 30ml seals the film wrapping, at 121 ℃ and 1.1kg.cm
-2Under the condition, with the high-pressure steam sterilizing pan 20min that sterilizes.The screening of condition of culture: in cultivation temperature is 25 ± 2 ℃, illumination 1500～2000LX, and every day, light application time was to cultivate in the culturing room of 12h.The result is referring to Fig. 1-2.
(2) will breed the aseptic seedling that obtains and transfer on the strong seedling culture base and carry out strong sprout, medium is with the wide-mouth bottle packing of 300ml, and every bottle of 30ml seals the film wrapping, at 121 ℃ and 1.1kg.cm
-2Under the condition, with the high-pressure steam sterilizing pan 20min that sterilizes.Condition of culture is with (1).The result is referring to Fig. 3.
(3) aseptic seedling is taken out from blake bottle, clean the root medium, and its base portion soaked in KIBA solution, in disposal plastic cup, a cup mesostroma is prepared by a certain percentage by yellow soil, charing rice husk ash, fine sand with the nursery stock cuttage, the rim of a cup use is sealed the film covering and is preserved moisture, cup is placed the management of taking root of the green house of tool intermittent spraying facility, and temperature keeps 20～30 ℃, 10 minutes intermittent times of spraying, spray time 10 seconds, rooting rate 87%.The result is referring to Fig. 4-5.
Embodiment 2
Clip is given birth to newly slightly then from the healthy and strong plant of oil tea, gets top 4-6cm disinfection; The aseptic seedling that first generation obtains is transferred to the WPM+BA7mg/L+IBA0.1mg/L+ZT0.05mg/L successive transfer culture, and be 40 days proliferating cycle, and growth coefficient reaches 7.0; Adopt WPM+IBA1.0mg/L+GA20mg/L to carry out strong sprout the aseptic seedling that obtains in the breeding, from blake bottle, take out when tissue cultivating seedling grew to 2-4cm in 25 days, its base portion is soaked 3s in the KIBA of 8000ppm solution, with the nursery stock cuttage in yellow soil, charing rice husk ash, fine sand by volume in the matrix of 1:2:1, rooting rate 89% after 40 days.
Concrete operations are referring to embodiment 1.
Embodiment 3
Clip is given birth to newly slightly then from the healthy and strong plant of oil tea, gets top 4-6cm disinfection; The aseptic seedling that first generation obtains is transferred to the WPM+BA5mg/L+IBA2.0mg/L+ZT2.0mg/L successive transfer culture, and be 35 days proliferating cycle, and growth coefficient reaches 5.2; Adopt WPM+IBA0.1mg/L+GA5mg/L to carry out strong sprout the aseptic seedling that obtains in the breeding, from blake bottle, take out when tissue cultivating seedling grew to 2-3cm in 23 days, its base portion is soaked 4s in the KIBA of 2000ppm solution, with the nursery stock cuttage in yellow soil, charing rice husk ash, fine sand by volume in the matrix of 1:3:1, rooting rate 94% after 30 days.
Concrete operations are referring to embodiment 1.
List of references:
[1] Rayleigh, the village. Chinese oil tea [M]. Beijing: China Forest publishing house, 1988
[2] Chen Yongzhong, poplar Xiao Hu, Peng Shaofeng. China's oil tea fine-variety breeding present Research and development tactics [J]. forestry science and technology exploitation, 2005,19 (4): 1-4
[3] Li Xiansheng. the development and use researchs [J] of China's oil tea resource. Hu'nan University of Science and Engineering's journal, 2005,26 (11): 127-129
[4] Yan Muqin. the generation [J] of oil tea somatic cell embryoid. Journal of Experimental Biology, 1980,3 (3): 343-347
[5] the grand hero that shakes. oil tea rataria cultured in vitro just obtains whole plant [J]. forestry science and technology communication, 1981 (1): 12-16
[6] Lu Tianling. the research [J] of oil tea unmature subleaf rataria cultured in vitro Cheng Miao. Journal of Experimental Biology, 1982,5 (4): 393-403
[7] the grand hero that shakes. the oil tea anther culture obtains green bud point and root [J]. forestry science and technology communication, 1981 (5): 6-9
[8] Bi Fangcheng, Tan Xiaofeng, Zhang Zhijun, etc. the research [J] of oil tea cultured in vitro regeneration induction plant. economic forest research, 2004,22 (2): 5-9
[9] Zhang Zhijun, Luo Shuping, Li Yaling, etc. Camellia Oleifera Clones cotyledon somatic embryo plant regeneration [J]. BULLETIN OF BOTANY Vol., 2005,22 (supplementary issue): 43-49
[10] Wu Zhengkai, Zheng Xiaofeng. oil tea tissue culture technique research [J]. Inner Mongol agricultural science and technology 2008 (1): 63-64
Claims (9)
1, a kind of clonal tissue culture of oil tea propagation method, comprise that just generation, subculture obtain tissue cultivating seedling, take root, transplant four-stage, it is characterized in that obtaining to take root behind the material, to take root and transplant step unite two into one the wherein preparation of the tissue cultivating seedling cuttage root-taking and the matrix of taking root: be mixed with the matrix of taking root in 1: 2 by volume～3: 1 by yellow soil, charing rice husk ash, fine sand; Matrix is packed in the cuttage container, with the tissue cultivating seedling cuttage wherein, and seal with light transmission film.
2, propagation method according to claim 1 after it is characterized in that tissue cultivating seedling taken out from blake bottle, carries out disinfection to tissue cultivating seedling.
3, propagation method according to claim 2 is characterized in that it being to soak 2-4 minute in 0.1% metalaxyl-MnZn solution.
4,, it is characterized in that the matrix of cuttage tissue cultivating seedling is also carried out disinfection according to claim 2 or 3 described propagation methods.
5,, it is characterized in that also the take root hormone preliminary treatment of material of tissue cultivating seedling according to the described propagation method of one of claim 1 to 3.
6, propagation method according to claim 5 is characterized in that the base portion of tissue cultivating seedling was soaked 3-4 seconds in the KIBA of 500～8000ppm solution.
7, according to the described propagation method of one of claim 1 to 3, it is characterized in that placing the green house of tool intermittent spraying facility to manage in the cuttage container of cuttage tissue cultivating seedling, temperature keeps 20～30 degree, after 30～40 days, the tissue cultivating seedling formation whole plant of taking root.
8,, it is characterized in that obtaining in the tissue cultivating seedling step according to the described propagation method of one of claim 1 to 3:
(1) subculture medium and hormone combination are that WPM+BA3-7mg/L+IBA0.1-2mg/L+ZT0.05-2mg/L breeds;
(2) strong seedling culture base and hormone combination are WPM+IBA0.1-1mg/L+GA5-20mg/L; Root induction.
9, the tissue cultivating seedling cuttage root-taking and the matrix of taking root is characterized in that being mixed with in 1: 2 by volume～3: 1 by yellow soil, charing rice husk ash, fine sand.
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101822160A (en) * | 2010-05-05 | 2010-09-08 | 广西壮族自治区林业科学研究院 | Method using tissue-subculture sprouts of oil-tea camellia to raise seedlings by grafting |
| CN102696480A (en) * | 2012-05-07 | 2012-10-03 | 广西壮族自治区林业科学研究院 | Method for culturing aseptic oil tea buds in test tube by using sand matrix |
| CN102860229A (en) * | 2012-08-05 | 2013-01-09 | 肖勇 | Tea tree tray-soilless seedling substrate |
| CN103039362A (en) * | 2013-01-14 | 2013-04-17 | 黄山市林业科学研究所 | Subculture medium for tissue culture seedling propagation of camellia oleifera abel and propagation method thereof |
| CN103461143A (en) * | 2013-09-30 | 2013-12-25 | 中南林业科技大学 | Method for tissue culture and rapid propagation of camellia oleifera |
| CN105557432A (en) * | 2016-01-07 | 2016-05-11 | 郎溪县姚村乡盛茂贵茶叶种植家庭农场 | Cuttage seedling raising method for promoting quick growth of yellow tea |
| CN107125136A (en) * | 2017-06-08 | 2017-09-05 | 合肥市风达农业有限责任公司 | A kind of Camellia nitidissima tissue culture mating system |
| CN107232002A (en) * | 2017-06-02 | 2017-10-10 | 金平县金山沉香油茶产销专业合作社 | A kind of oil tea cuttage planting method |
| CN107278561A (en) * | 2017-06-28 | 2017-10-24 | 玉屏侗族自治县油茶产业发展领导小组办公室 | A kind of seed seedling-raising method of oil tea |
| CN108402082A (en) * | 2018-02-28 | 2018-08-17 | 湖南省林业科学院 | The root-growing agent and its rooting method for promoting oil tea nursery stock to take root |
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| CN101822160A (en) * | 2010-05-05 | 2010-09-08 | 广西壮族自治区林业科学研究院 | Method using tissue-subculture sprouts of oil-tea camellia to raise seedlings by grafting |
| CN102696480A (en) * | 2012-05-07 | 2012-10-03 | 广西壮族自治区林业科学研究院 | Method for culturing aseptic oil tea buds in test tube by using sand matrix |
| CN102860229A (en) * | 2012-08-05 | 2013-01-09 | 肖勇 | Tea tree tray-soilless seedling substrate |
| CN102860229B (en) * | 2012-08-05 | 2013-11-06 | 肖勇 | Tea tree tray-soilless seedling substrate |
| CN103039362A (en) * | 2013-01-14 | 2013-04-17 | 黄山市林业科学研究所 | Subculture medium for tissue culture seedling propagation of camellia oleifera abel and propagation method thereof |
| CN103461143A (en) * | 2013-09-30 | 2013-12-25 | 中南林业科技大学 | Method for tissue culture and rapid propagation of camellia oleifera |
| CN105557432A (en) * | 2016-01-07 | 2016-05-11 | 郎溪县姚村乡盛茂贵茶叶种植家庭农场 | Cuttage seedling raising method for promoting quick growth of yellow tea |
| CN107232002A (en) * | 2017-06-02 | 2017-10-10 | 金平县金山沉香油茶产销专业合作社 | A kind of oil tea cuttage planting method |
| CN107125136A (en) * | 2017-06-08 | 2017-09-05 | 合肥市风达农业有限责任公司 | A kind of Camellia nitidissima tissue culture mating system |
| CN107278561A (en) * | 2017-06-28 | 2017-10-24 | 玉屏侗族自治县油茶产业发展领导小组办公室 | A kind of seed seedling-raising method of oil tea |
| CN108402082A (en) * | 2018-02-28 | 2018-08-17 | 湖南省林业科学院 | The root-growing agent and its rooting method for promoting oil tea nursery stock to take root |
| CN108402082B (en) * | 2018-02-28 | 2021-07-06 | 湖南省林业科学院 | Rooting agent for promoting rooting of camellia oleifera seedlings and rooting method thereof |
| CN113966716A (en) * | 2021-06-17 | 2022-01-25 | 陕西理工大学 | Camellia oleifera callus in-vitro culture and karyotype analysis method |
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