CN101027975A - Fast reproduction of European wild black cherry tissue - Google Patents
Fast reproduction of European wild black cherry tissue Download PDFInfo
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- CN101027975A CN101027975A CNA2007100678429A CN200710067842A CN101027975A CN 101027975 A CN101027975 A CN 101027975A CN A2007100678429 A CNA2007100678429 A CN A2007100678429A CN 200710067842 A CN200710067842 A CN 200710067842A CN 101027975 A CN101027975 A CN 101027975A
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Abstract
A tissue culture method for fast reproduction of wild European cherry includes such steps as choosing explant, sterilizing it, removing two brocon ends, inoculating it in the inductive culture medium, inductive culture to induce terminal bud and axillary bud, differentiating rosette buds, cutting the rosette buds to obtain single bud, reproductive culture in the reproducing culture medium, rooting culture in the culture medium A and then in culture medium B to obtain tissue cultured seedlings, naturalizing, and transplanting them in the matrix prepared from peat, pearlite and vermiculite.
Description
Technical field
The present invention relates to the woody plant tissure culture technique, mainly is a kind of method of European wild black cherry mahogany tissue-culturing rapid propagation.
Background technology
Europe wild black cherry mahogany (Wild Cherry, Prunus Avium wildstar) is the cultivar of cultivating in the wild morello wood that Britain finds, these seeds are unique seeds that have commercial with material value during cherry belongs to.These seeds are high megaphanerophyte, and the height of tree reaches as high as 39 meters, and the maximum diameter of a cross-section of a tree trunk 1.3 meters above the ground is 2.5 meters, and trunk is perfectly straight satisfactory, major branch rareness, trunk is liftoff 3～6 meters high no branches.The lumber quality height, timber light weight, solid, texture is straight and fine and closely woven, and color is pale red to be the high-quality material of doing noble furniture and plywood veneer to pale red brown, and economic worth is very high, 1500～2000 dollars every cubic metre of original producton location sheet material prices.It also is widely used in aircraft, boats and ships, military project furniture, engraving, floor etc.Except that as high-quality with the material, cherry also can do ornamental tree species, medicinal seeds, has many-sided quality merchandise value.The nearly all up to now broad-leaved tree of Britain all is to use seminal propagation, seed is actually the result of two parents hybridization, so they have produced a large amount of variations, and some are desirable, some are undesirable, so their phenotype has very large difference.
The academy of agricultural sciences, Zhejiang Province introduces European wild black cherry mahogany kind from Britain, intends breeding seedling in a large number by plant tissue culture technique, exports American-European countries then.Promoted after can also planting experimentally at home simultaneously.Adopt method for plant tissue culture can breed various plants in a short time fast, reproduction rate height not only, and, can keep the merit of former stock because it is vegetative propagation, on producing, use more and more wider in recent years.But the complexity of different plant utilization method for plant tissue culture breeding seedling is different, and particularly woody plant is difficult.In group training test in the early stage of European wild black cherry mahogany, woody plant normal some problems that occur in tissue culture have also appearred, as explant inoculation pollution rate height, the easy vitrifying of tissue cultivating seedling, yellow, take root problems such as difficulty, transplanting survival rate be low of tissue cultivating seedling make it can't realize large-scale industrialized production.
Summary of the invention
The present invention will solve the defective of above-mentioned prior art, and a kind of method of European wild black cherry mahogany tissue-culturing rapid propagation is provided.
The present invention solves the technical scheme that its technical problem adopts.
The method of this European wild black cherry mahogany tissue-culturing rapid propagation, carry out as follows:
1), explant selection and sterilization: the semi-lignified spray of the potted plant wild black cherry mahogany of clip seedling cuts its terminal bud and the stem section of being with axillalry bud, after surface sterilizing is handled, as the explant of group training usefulness;
2), inducing culture: will be behind an end or the excision of two end-grain cutting mouth browning places of the explant terminal bud behind the surface sterilizing and the stem section of band axillalry bud under aseptic condition, the bud of terminal bud and stem segment with axillary bud upwards is seeded on the inducing culture, induce terminal bud and axillalry bud rudiment, then differentiation clump bud;
3), enrichment culture: under aseptic condition, the clump bud is cut into individual plant and inserts in the proliferated culture medium and carry out enrichment culture, the induced bundle bud;
4), culture of rootage: the tissue culture plant inoculation that earlier growth is reached 2～3 centimetres was cultivated 7 days in root media 1, changed in the root media 2 again and cultivated, and induced it to take root;
5), the domestication and the transplanting of tissue cultivating seedling: when the tissue cultivating seedling growth reaches more than 3～5 centimetres, carry out the domestication of tissue cultivating seedling earlier, be transplanted into again in the mixed-matrix of peat, perlite, vermiculite.
The tissue-culturing rapid propagation medium that the present invention adopts, by minimal medium and do not train the medium in stage on the same group and form, minimal medium and each stage medium additives content are:
1), to select MS+ sucrose or white sugar 30g/L+ agar for use be 7g/L for the minimal medium of inducing culture and enrichment culture, pH5.8; It is 7g/L for 20g/L+ agar that the minimal medium of culture of rootage is selected 1/2MS+ sucrose or white sugar for use, pH5.8;
2), inducing culture: MS+BA0.25～1.0mg/L+NAA0.05～0.5mg/L;
3), proliferated culture medium: MS+BA0.25～1.0mg/L+NAA0.005～0.05mg/L;
4), root media 1:1/2MS+IBA1～3mg/L, root media 2:1/2MS.
The tissue-culturing rapid propagation medium that the present invention adopts, by minimal medium and do not train the medium in stage on the same group and form, minimal medium and each stage medium additives content are:
1), to select MS+ white sugar 30g/L+ agar for use be 7g/L for the minimal medium of inducing culture and enrichment culture, pH5.8; It is 7g/L for 20g/L+ agar that the minimal medium of culture of rootage is selected 1/2MS+ white sugar for use, pH5.8;
2), inducing culture: MS+BA0.75mg/L+NAA0.1mg/L;
3), proliferated culture medium: MS+BA0.5mg/L+NAA0.01mg/L;
4), root media 1:1/2MS+IBA2mg/L, root media 2:1/2MS.
The condition of culture of respectively organizing the training stage that the present invention adopts is: in inducing culture and enrichment culture stage: 22 ± 2 ℃ of temperature, illumination 12h/d, intensity of illumination are about 3000lx; In the culture of rootage stage: 20 ± 2 ℃ of temperature, illumination 8h/d, intensity of illumination are about 2000lx.
The transplanting medium that the present invention adopts is by peat: perlite: vermiculite 2: 1: 1 by volume is formulated.
The effect that the present invention is useful is:
1), set up European wild black cherry mahogany group culturation rapid propagating technology system, inducing culture, proliferated culture medium and the root media of suitable European wild black cherry mahogany tissue-culturing rapid propagation have been adopted, month rate of increase can reach 3 times, tissue cultivating seedling does not have vitrifying and yellow, rooting rate reaches 100%, and tissue cultivating seedling robust growth, leaf look dark green, and transplanting survival rate can reach 95%, thereby key technologies such as explant sterilization is difficult for, the easy vitrifying of tissue cultivating seedling, yellow, rooting and transplant difficulty have been solved, for batch production production is laid a good foundation.
2), the European wild black cherry mahogany tissue cultivating seedling that obtains of the quick breeding method for tissue culture that proposes, the genetic character unanimity has overcome conventional seminal propagation offspring Yi Bianyi, shortcoming that the cottage propagation coefficient is low.
3), utilize the European wild black cherry mahogany tissue cultivating seedling of plant tissue culture technique breeding not only can export goods and earn foreign currency, also can be applicable at home, will produce good economic society ecological benefits.
4), applied for invention patent protection, not only help to change product image in the international market, but also help to improve the occupation rate of market of exported product.
Embodiment
The invention will be described further below in conjunction with embodiment: but content of the present invention is not limited thereto.
Embodiment 1: this European wild black cherry mahogany tissue culture and rapid propagation method, and concrete steps are as follows:
1), explant selection and sterilization: the semi-lignified spray of the potted plant wild black cherry mahogany of clip seedling, cut the stem section of its terminal bud and band axillalry bud, explant is through 75% alcohol-pickled 0.5～1.0min, with volume ratio is 2% aqueous sodium hypochlorite solution sterilization, 10～15min, and aseptic water washing is 3～5 times then.After surface sterilizing is handled, as the explant of group training usefulness.
2), inducing culture: will be behind an end or the excision of two end-grain cutting mouth browning places of the explant terminal bud behind the surface sterilizing and the stem section of band axillalry bud under aseptic condition, the bud of terminal bud and stem segment with axillary bud upwards is seeded on the inducing culture of MS+BA0.25～1.0mg/L+NAA0.05～0.5mg/L, induce terminal bud and axillalry bud rudiment, differentiate clump bud then.
3), enrichment culture: under aseptic condition, the clump bud is cut into again in the proliferated culture medium that individual plant inserts MS+BA0.25～1.0mg/L+NAA0.005～0.05mg/L and carries out enrichment culture, induce clump bud.Per 6 all shoot proliferations are cultivated 1 time.
4), culture of rootage: the tissue culture plant inoculation that earlier growth is reached 2～3 centimetres was cultivated 7 days in the root media 1 of 1/2MS+IBA1～3mg/L, and changing 1/2MS again over to does not have cultivation in the hormone root media 2, induces it to take root.
5), the domestication and the transplanting of tissue cultivating seedling: when the tissue cultivating seedling growth reaches more than 3～5 centimetres, carry out the domestication of tissue cultivating seedling earlier, the volume ratio that is transplanted into peat, perlite, vermiculite again is in the nutritive cube of 2: 1: 1 matrix.
Each condition of culture of organizing the training stage is: in inducing culture and enrichment culture stage: 22 ± 2 ℃ of temperature, illumination 12h/d, intensity of illumination are about 3000lx; In the culture of rootage stage: 20 ± 2 ℃ of temperature, illumination 8h/d, intensity of illumination are about 2000lx.
Embodiment 2: this European wild black cherry mahogany tissue culture and rapid propagation method, and concrete steps are as follows:
1), explant selection and sterilization: the semi-lignified spray of the potted plant wild black cherry mahogany of clip seedling, cut the stem section of its terminal bud and band axillalry bud, explant is through 75% alcohol-pickled 0.5～1.0min, with volume ratio is 2% aqueous sodium hypochlorite solution sterilization, 10～15min, and aseptic water washing is 3～5 times then.Handle through surface sterilizing, as the explant of group training usefulness.
2), inducing culture: will be behind an end or the excision of two end-grain cutting mouth browning places of the explant terminal bud behind the surface sterilizing and the stem section of band axillalry bud under aseptic condition, the bud of terminal bud and stem segment with axillary bud upwards is seeded on the inducing culture of MS+BA0.75mg/L+NAA0.1mg/L, induce terminal bud and axillalry bud rudiment, differentiate clump bud then.
3), enrichment culture: under aseptic condition, the clump bud is cut into again in the proliferated culture medium that individual plant inserts MS+BA0.5mg/L+NAA0.01mg/L and carries out enrichment culture, induce clump bud.Per 6 all shoot proliferations are cultivated 1 time.
4), culture of rootage: the tissue culture plant inoculation that earlier growth is reached 2～3 centimetres was cultivated 7 days in the root media 1 of 1/2MS+IBA2mg/L, and changing 1/2MS again over to does not have cultivation in the hormone root media 2, induces it to take root.
5), the domestication and the transplanting of tissue cultivating seedling: when the tissue cultivating seedling growth reaches more than 3～5 centimetres, carry out the domestication of tissue cultivating seedling earlier, the volume ratio that is transplanted into peat, perlite, vermiculite again is in the nutritive cube of 2: 1: 1 matrix.
Each condition of culture of organizing the training stage is: in inducing culture and enrichment culture stage: 22 ± 2 ℃ of temperature, illumination 12h/d, intensity of illumination are about 3000lx; In the culture of rootage stage: 20 ± 2 ℃ of temperature, illumination 8h/d, intensity of illumination are about 2000lx.
Claims (5)
1, a kind of method of European wild black cherry mahogany tissue-culturing rapid propagation is characterized in that:
1.1), explant selection and sterilization: the semi-lignified spray of the potted plant wild black cherry mahogany of clip seedling cuts the stem section of its terminal bud and band axillalry bud, after surface sterilizing is handled, as the explant of group training usefulness;
1.2), inducing culture: will be behind the end of stem section or the excision of two end-grain cutting mouth browning places of explant terminal bud behind the surface sterilizing and band axillalry bud under aseptic condition, the bud of terminal bud and stem segment with axillary bud upwards is seeded on the inducing culture, induce terminal bud and axillalry bud rudiment, then differentiation clump bud;
1.3), enrichment culture: under aseptic condition, the clump bud is cut into individual plant and inserts in the proliferated culture medium and carry out enrichment culture, the induced bundle bud;
1.4), culture of rootage: the tissue culture plant inoculation that earlier growth is reached 2～3 centimetres was cultivated 7 days in root media 1, changed in the root media 2 again and cultivated, and induced it to take root;
1.5), the domestication and the transplanting of tissue cultivating seedling: when the tissue cultivating seedling growth reaches more than 3～5 centimetres, carry out the domestication of tissue cultivating seedling earlier, be transplanted into again in the mixed-matrix of peat, perlite, vermiculite.
2, the method for European wild black cherry mahogany tissue-culturing rapid propagation according to claim 1, it is characterized in that, the medium of described tissue-culturing rapid propagation comprises minimal medium and does not train the medium in stage on the same group, and minimal medium and each stage medium additives content are:
2.1), to select MS+ sucrose or white sugar 30g/L+ agar for use be 7g/L for the minimal medium of inducing culture and enrichment culture, pH5.8; It is 7g/L for 20g/L+ agar that the minimal medium of culture of rootage is selected 1/2MS+ sucrose or white sugar for use, pH5.8;
2.2), inducing culture: MS+BA0.25～1.0mg/L+NAA0.05～0.5mg/L;
2.3), proliferated culture medium: MS+BA0.25～1.0mg/L+NAA0.005～0.05mg/L;
2.4), root media 1:1/2MS+IBA1～3mg/L, root media 2:1/2MS.
3, the method for a kind of European wild black cherry mahogany tissue-culturing rapid propagation according to claim 2, it is characterized in that, the medium of described tissue-culturing rapid propagation comprises minimal medium and does not train the medium in stage on the same group, and minimal medium and each stage medium additives content are:
3.1), to select MS+ white sugar 30g/L+ agar for use be 7g/L for the minimal medium of inducing culture and enrichment culture, pH5.8; It is 7g/L for 20g/L+ agar that the minimal medium of culture of rootage is selected 1/2MS+ white sugar for use, pH5.8;
3.2), inducing culture: MS+BA0.75mg/L+NAA0.1mg/L;
3.3), proliferated culture medium: MS+BA0.5mg/L+NAA0.01mg/L;
3.4), root media 1:1/2MS+IBA2mg/L, root media 2:1/2MS.
4, according to the method for claim 1 or 2 or 3 described European wild black cherry mahogany tissue-culturing rapid propagations, it is characterized in that the described condition of culture of respectively organizing the training stage is: in inducing culture and enrichment culture stage: 22 ± 2 ℃ of temperature, illumination 12h/d, intensity of illumination are about 3000lx; In the culture of rootage stage: 20 ± 2 ℃ of temperature, illumination 8h/d, intensity of illumination are about 2000lx.
5, the method for European wild black cherry mahogany tissue-culturing rapid propagation according to claim 1 is characterized in that, described transplanting medium is by peat: perlite: vermiculite 2: 1: 1 by volume is formulated.
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Cited By (8)
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| CN102657098A (en) * | 2012-06-05 | 2012-09-12 | 上海闵行区苗圃 | In-vitro culture method for tender stem segments of cherry rootstock |
| CN103299909A (en) * | 2013-07-08 | 2013-09-18 | 上海闵行区苗圃 | Method for breeding cherry seedlings in large scales through tissue culturing |
| CN104012406A (en) * | 2014-04-04 | 2014-09-03 | 大连大学 | Regeneration in-vitro method for sweet cherry variety wanhongzhu |
| CN104521755A (en) * | 2014-12-18 | 2015-04-22 | 山东省果树研究所 | Culture medium for preventing etiolation of Yunan wild cherry test-tube plantlet and tissue culture method |
| CN105494100A (en) * | 2015-12-30 | 2016-04-20 | 江汉大学 | Tissue culture and rapid propagation method of oriental cherries |
| CN109362524A (en) * | 2018-11-07 | 2019-02-22 | 中国科学院昆明植物研究所海盐工程技术中心 | A kind of cultural method of African Chrysanthemum new varieties |
| CN110463607A (en) * | 2019-09-09 | 2019-11-19 | 新疆农业大学 | The method of wild Europe Lee's leaf tissue culture |
| CN111084046A (en) * | 2019-12-24 | 2020-05-01 | 河北理查德农业科技有限公司 | Cherry breeding method |
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| JP4647804B2 (en) * | 2000-02-22 | 2011-03-09 | 住友林業株式会社 | A method for mass growth of cherry genus by tissue culture |
| CN1399868A (en) * | 2001-08-02 | 2003-03-05 | 初慧民 | Base material for cultivating plant |
| CN1285259C (en) * | 2004-12-27 | 2006-11-22 | 华南师范大学 | Breeding method of Wedelia trilobata |
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- 2007-03-29 CN CNB2007100678429A patent/CN100553446C/en not_active Expired - Fee Related
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102657098A (en) * | 2012-06-05 | 2012-09-12 | 上海闵行区苗圃 | In-vitro culture method for tender stem segments of cherry rootstock |
| CN103299909A (en) * | 2013-07-08 | 2013-09-18 | 上海闵行区苗圃 | Method for breeding cherry seedlings in large scales through tissue culturing |
| CN103299909B (en) * | 2013-07-08 | 2014-10-01 | 上海闵行区苗圃 | Method for breeding cherry seedlings in large scales through tissue culturing |
| CN104012406A (en) * | 2014-04-04 | 2014-09-03 | 大连大学 | Regeneration in-vitro method for sweet cherry variety wanhongzhu |
| CN104012406B (en) * | 2014-04-04 | 2016-08-17 | 大连大学 | The in-vitro regeneration method of sweet cherry variety red pearl in evening |
| CN104521755A (en) * | 2014-12-18 | 2015-04-22 | 山东省果树研究所 | Culture medium for preventing etiolation of Yunan wild cherry test-tube plantlet and tissue culture method |
| CN105494100A (en) * | 2015-12-30 | 2016-04-20 | 江汉大学 | Tissue culture and rapid propagation method of oriental cherries |
| CN109362524A (en) * | 2018-11-07 | 2019-02-22 | 中国科学院昆明植物研究所海盐工程技术中心 | A kind of cultural method of African Chrysanthemum new varieties |
| CN109362524B (en) * | 2018-11-07 | 2020-11-17 | 中国科学院昆明植物研究所海盐工程技术中心 | Cultivation method of new gerbera jamesonii variety |
| CN110463607A (en) * | 2019-09-09 | 2019-11-19 | 新疆农业大学 | The method of wild Europe Lee's leaf tissue culture |
| CN111084046A (en) * | 2019-12-24 | 2020-05-01 | 河北理查德农业科技有限公司 | Cherry breeding method |
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