CN1344489A - Fast excised African chrysanthemum propagation method - Google Patents
Fast excised African chrysanthemum propagation method Download PDFInfo
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- CN1344489A CN1344489A CN 01129821 CN01129821A CN1344489A CN 1344489 A CN1344489 A CN 1344489A CN 01129821 CN01129821 CN 01129821 CN 01129821 A CN01129821 A CN 01129821A CN 1344489 A CN1344489 A CN 1344489A
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Abstract
The fast excised propagation of African chrysanthemum includes three improved stpes: the disinfection of explant, induced germination of adventitious bud and culture to root. In the disinfection of explant, bud is first treated of low temperature before the surface disinfection of receptacle; in induced germination of adventitious bud, culture medium with phenyl carbamide as cytomin is used; and in culture to root, test tube seedling base part is soaked with auxin solution before culture in solid culture medium containing no hormone. The said method can raise the asexual propagation coefficient of African chrysanthemum through the simple operate process.
Description
The present invention relates to a kind of method for plant tissue culture, specifically, be meant a kind of tissue culture technique method that cultivation is bred fast to excised African chrysanthemum that adopts.
Plant Tissue Breeding is meant any organ, tissue or the cell with plant, carries out the process that aseptic culture is grown under the manual control condition.This technology is drawn materials few, and it is low to cultivate the vegetable material cost, and growth cycle is short, convenient management.Utilize this quick propagating technology of Plant Tissue Breeding, can multiply the plant of maternal biological nature of a large amount of maintenances and genetic character at short notice.It is reported that China plant of breeding fast has nearly dried kind.Normally used minimal medium has several, as MS, B5, White, N6 etc.In tissue culture, plant explants will be under the effect of plant growth substance, through inducing dedifferentiation, recover fissional ability, influencing processes such as breaking up, promote organ formation again.The size of plant explants reproduction coefficient directly affects the quality and quantity of producing the offspring plant.
Flameray gerbera (Gerbera jamesonil has another name called African daisy) belongs to the African Chrysanthemum in the composite family, and nearly in the world at present kind more than 80 is that a kind of ornamental value is higher, the fresh cut-flowers of domestic and international fast sale and potted plant famous-brand and high-quality flowers, the strains up to a million of annual market demand.But flameray gerbera is a cross-pollinated plant, seed progeny's life-span is short, germination rate is low and aberration rate is high, can not keep all property of original, simultaneously division propagation coefficient low (every strain only divides 5～6 strains), and degenerate easily and propagate disease, so conventional breeding chrysanthemum method is difficult to carry out large-scale production.[Ru Huangji is bright, Ni Yueyuan, Lin Manhong for the existing at home report of the method for tissue culture of relevant flameray gerbera vegetative propagation seedling.The quick breeding of flameray gerbera.The gardening journal, 1987,14 (1): 125; The Shandong Setsuka, woods is brave, Guo Wenjie.The cultured in vitro of the little holder of flameray gerbera.The subtropical plant communication, 1996,25 (2): 21; Liu Qunliang, Huang Aibing, Li Hua is foster etc.Flameray gerbera tissue culture quick breeding Study on Technology.Guangdong gardening, 2001,2 (1): 21 etc.], its method generally comprises following steps: (1) explant sterilization: be explant successively adopt 75% ethanol and the surface sterilization of 0.1-0.2% mercuric chloride each 1 minute and 10-20 minute (add surfactant number droplet) with the holder; (2) evoking adventive bud takes place: will be cut into the square of 0.3-1 centimeter length through the explant of surface sterilization, being placed in 1/2 MS, B5 or the White solid culture medium 25-40 days respectively takes place with evoking adventive bud, should add the benzyl purine class basic element of cell division of 0.1-10mg/L and indoles or the naphthalene class growth hormone of 0.1-2mg/L in the medium, its condition of culture is: 25 ± 2 ℃ of temperature, intensity of illumination 2000 ± 50Lx, light application time 12-16 hour/day; (3) cultivation is taken root: the indefinite bud that will highly be 3-7 centimetre cuts into individual plant, (indefinite bud that generally adopts 1/2MS to incite somebody to action substantially highly to 3-7 centimetre cuts into individual plant at the solid culture medium of taking root, go up cultivation at the solid culture medium of taking root (generally adopting the 1/2MS minimal medium), the indoles or the naphthalene class growth hormone that should add 0.1-5mg/L in the medium, the same step of condition of culture (2) can produce by inducing adventitious root in 25-50 days.Because flameray gerbera is wide in variety, it is very big to induce the capacity variance of indefinite bud by holder between each kind, and the bud ratio of directly being induced by holder is extremely low, so this method is unfavorable for the quick breeding of flameray gerbera equally.
In regulating controlling plant material cultured in vitro process, outside the Pass its reproduction coefficient removes and to have with condition of culture such as the characteristic of vegetable material itself and nutrition, illumination, temperature, plant growth substance also plays an important role, and wherein influence is the most significant is auxins and cytokinin-like substance.Other has report to point out, before the bud to woody plant carried out cultured in vitro, carrying out low temperature preliminary treatment a period of time was of value to cultured in vitro growth (the yellow-study woods, Li Xiaoju writes.Higher plant is organized the morphogenesis and the regulation and control thereof of cultured in vitro, Beijing: Science Press, 1995,78), but the present identical report that the herbaceous plant aspect is not arranged.
The objective of the invention is to the deficiency at the existing method for tissue culture of flameray gerbera, a kind of new method of quick breeding is provided, it can obviously improve the vegetative propagation coefficient of flameray gerbera, and easy and simple to handle.
In order to realize purpose of the present invention, the present invention carries out following improvement on the basis of existing method for tissue culture: in step (1), before the holder surface sterilization, at first bud is carried out low temperature treatment.
Further improvement of the present invention is: on the basis of low temperature treatment, the medium in the step (2) adopts the phenyl ureas basic element of cell division to replace the benzyl purine class basic element of cell division, to improve the ability that evoking adventive bud takes place.
Better scheme of the present invention is: on above-mentioned improved basis of two steps, in step (3), change the method that auxin substance is handled test-tube plantlet simultaneously, to shorten the time that the test-tube plantlet adventive root takes place, that is: at first with indoles or naphthalene class growth hormone solution soaking test-tube plantlet base portion, and then change on the 1/2MS solid culture medium that does not contain hormone and cultivate.
The specific requirement of this method is as follows: temperature range is 9 ± 2 ℃ in the low temperature treatment, and the processing time is 1-5 days, preferably 3 days; In the medium that step (2) adopts, the phenyl ureas basic element of cell division can adopt 2-chloro-4-pyridine radicals-N-phenylurea (being called for short CPPU), (4-pyrans)-1,1-purine amino-9-hydrogen pyranose-9H purine-4-hydrogen pyranose (being called for short TDZ) etc., preferably adopt 2-chloro-4-pyridine radicals-N-phenylurea (being called for short CPPU), its content in minimal medium is: 0.5-2mg/L, preferably 1mg/L; In the step (3), indoles growth hormone can adopt heteroauxin, indolebutyric acid etc., and naphthalene class growth hormone adopts methyl.Auxin substance solution content is 2mg/L-15mg/L, and its preferable range is 5-10mg/L, and the time that the test-tube plantlet base portion soaks is 1 day, and the continuation incubation time in solid culture medium is 5-15 days.
Because the present invention adopts the low temperature preprocess method, promoted the cultured in vitro growth of flameray gerbera, further adopt the phenyl ureas basic element of cell division again, improved the ability that evoking adventive bud takes place, also change auxin substance and handle time, the raising adventive root quality that the method for test-tube plantlet takes place to shorten the test-tube plantlet adventive root, so improved the in-vitro propagate coefficient of flameray gerbera widely, help the quick breeding of flameray gerbera, reached purpose of the present invention.
The present invention has following advantage and effect:
1, the experiment proved that, with conventional method relatively, before inoculation, bud carried out 9 ± 2 ℃ of low temperature treatment of 1～5 day, cut holder and cultivated 30 days, the bud number average that the induction frequency of indefinite bud and every explant produce is than conventional method height; In the step (2), adopt the phenyl ureas basic element of cell division to replace the benzyl purine class basic element of cell division, the concentration in the medium is lowered, adventitious bud induction frequency improves; Improve the concentration of auxin substance in the root media, adopt infusion method to handle, the adventive root time of origin shortens, and rooting rate increases, and the number of taking root of every strain test-tube plantlet is improved, and grows fine, and has guaranteed test-tube plantlet adventive root quality.
2, the inventive method is compared with existing conventional method, need not to increase special equipment, and is easy and simple to handle.
Below in conjunction with embodiment the present invention is further detailed:
Embodiment 1:
Get 42 on the orange red kind flameray gerbera holder of clearing away heart-fire as explant, at first bud is placed 9 ± 2 ℃ environment to handle 5 days, take out holder according to conventional method then and carry out surface sterilization and inoculated and cultured, its medium is 1/2MS, contain 0.5mg/L heteroauxin and 10mg/L 6-benzyladenine, find when cultivating 30 days, existing 18 explants germinate, adventitious bud induction frequency reaches 41.9%, and on average 3.3 buds take place each explant.The cutting indefinite bud is cultivated into seedling, obtains healthy seedling, and the growth shape of its adventive root is better.And adopt conventional method to cultivate the control group of (without low temperature treatment), and the incidence of its indefinite bud only is 22.2%, and it is few that its adventive root takes place, and growth is slow, obviously is worse than the inventive method.
Embodiment 2:
Other operation and control group are with embodiment 1, the orange red explant that clears away heart-fire is 56, the time of low temperature treatment bud is 3 days, cultivating had 24 explants to germinate in 30 days, bud induction rate reaches 42.9%, and on average each explant has 6 buds to form, and takes root through cultivation, the upgrowth situation of its adventive root is better, and adventitious bud induction frequency and adventive root upgrowth situation all obviously are better than control group.
Embodiment 3:
Other operation and control group are with embodiment 1, and flameray gerbera adopts the bright red kind that clears away heart-fire, and the low temperature treatment time is 1 day, cultivate 30 days indefinite bud incidences and reach 14.3%, 2 of the flower bud number that average each holder explant forms are taken root through cultivation, and the upgrowth situation of its adventive root is general.And the adventitious bud induction frequency of control group only is 5%, and on average the flower bud number of each holder explant formation is 1, and the growth difficulty of adventive root obviously is worse than the inventive method.
Embodiment 4:
Other is with embodiment 3, and flameray gerbera adopts the bright red kind that clears away heart-fire, and the medium of step (2) is 1/2 B5, cultivates 30 days indefinite bud incidences and reaches 12.5%, and on average the flower bud number of each holder explant formation is 2, and the upgrowth situation of its adventive root is general.And the adventitious bud induction frequency of control group only is 5.8%, and on average the flower bud number of each holder explant formation is 1, and difficulty takes place adventive root, and established adventive root poor growth obviously is worse than the inventive method.
Embodiment 5:
Other is with embodiment 3, and the medium of step (2) is 1/2 White, cultivates 30 days indefinite bud incidences and reaches 13.0%, and on average the flower bud number of each holder explant formation is 1.6, has adventive root to take place, and growth conditions is general.And the adventitious bud induction frequency of control group only is 5.8%, and on average the flower bud number of each holder explant formation is 1, and difficulty takes place adventive root, and established adventive root poor growth obviously is worse than the inventive method.
Embodiment 6:
Other is with embodiment 1, flameray gerbera adopts the orange red kind that clears away heart-fire, the time of low temperature treatment is 3 days, the medium that adopts in the step (2) is 1/2MS, and wherein the basic element of cell division adopts 2-chloro-4-pyridine radicals-N-phenylurea (being called for short CPPU) of 1mg/L, and adventitious bud induction frequency is 43% as a result, cultivate through step (3), the adventive root upgrowth situation is good, and it is many that adventive root produces quantity, and growth is fast and sturdy.And the control group of employing conventional method (do not have low temperature treatment, the basic element of cell division is the 10mg/L 6-benzyladenine), its adventitious bud induction frequency is 34.7%, the adventive root upgrowth situation is relatively poor, obviously is worse than the inventive method.
Embodiment 7:
Other is with embodiment 6, flameray gerbera adopts the orange red kind of evil mind, the time of low temperature treatment is 3 days, the basic element of cell division adopts 2-chloro-4-pyridine radicals-N-phenylurea (being called for short CPPU) of 0.5mg/L, its adventitious bud induction frequency is 20% as a result, and the adventive root upgrowth situation is better, and the adventitious bud induction frequency of control group is 12.4%, the adventive root upgrowth situation a little less than, both all obviously are worse than the inventive method.
Embodiment 8:
Other is with embodiment 6, flameray gerbera adopts the orange red kind of evil mind, the time of low temperature treatment is 3 days, the basic element of cell division adopts 2-chloro-4-pyridine radicals-N-phenylurea (being called for short CPPU) of 2mg/L, its adventitious bud induction frequency is 22.2% as a result, and the adventive root upgrowth situation is better, and the adventitious bud induction frequency of control group is 8.3%, the adventive root upgrowth situation a little less than, both all obviously are worse than the inventive method.
Embodiment 9:
Other is with embodiment 8, flameray gerbera adopts the orange red kind of evil mind, the time of low temperature treatment is 3 days, the basic element of cell division adopts (the 4-pyrans)-1 of 2mg/L, 1-purine amino-9-hydrogen pyranose-9H purine-4-hydrogen pyranose (being called for short TDZ), and its adventitious bud induction frequency is 20.3% as a result, the adventive root upgrowth situation is better, and the adventitious bud induction frequency of control group is 8.3%, the adventive root upgrowth situation a little less than, both all obviously are worse than the inventive method.
Embodiment 10:
Other is with embodiment 6, flameray gerbera adopts the pink kind that clears away heart-fire, soaked the test-tube plantlet base portion 1 day with the 2mg/L naphthalene acid solution in the step (3), continue then to cultivate in the 1/2MS solid culture medium, its root induction rate was 50% in 15 days as a result, root growth is good, and three steps all adopt the control group of conventional method, and its root induction rate is 20%, and root takes place seldom, it is extremely slow to grow, and obviously is worse than the inventive method.
Embodiment 11:
Other is with embodiment 10, use 15mg/L heteroauxin solution soaking test-tube plantlet base portion 1 day in the step (3), cultivating 15 days its root induction rates is 88%, the adventive root growth better, and three steps all adopt the control group of conventional method, and its root induction rate is 20%, and adventive root takes place few, root growth is slow, obviously is worse than the inventive method.
Embodiment 12:
Other uses 5mg/L indolebutyric acid solution soaking test-tube plantlet base portion 1 day with embodiment 10 in step (3), continue then to cultivate in the 1/2MS solid culture medium, cultivating 15 days its root induction rates is 93%, and adventive root takes place many, and the radical of every strain test-tube plantlet growth has 2, root growth is good, and fibrous root is arranged.And three steps all adopt the control group of conventional method, and its root induction rate is 18%, and adventive root takes place few, and root growth is slow, obviously is worse than the inventive method.
Embodiment 13:
Other is with embodiment 10, flameray gerbera adopts the pink kind that clears away heart-fire, and uses 10mg/L indolebutyric acid solution soaking test-tube plantlet base portion 1 day in the step (3), continues then to cultivate 15 days in the 1/2MS solid culture medium, its root induction rate is 100%, and the radical of every strain test-tube plantlet growth has 2.2.Root is long, grows, and fibrous root is many, and the strong leaf of seedling is green.And three steps all adopt the control group of conventional method, and its root induction rate is 34%, and every strain rooting of vitro seedling number is 1.3, and root is carefully short and small, and fibrous root is few, and growth is slow, obviously is worse than the inventive method.
Claims (5)
1, a kind of method of fast excised African chrysanthemum propagation, three steps that comprise that explant sterilization, evoking adventive bud take place and cultivation is taken root is characterized in that: in step (1), before the holder surface sterilization, at first bud is carried out low temperature treatment.
2, the method for claim 1 is characterized in that: the basic element of cell division that used medium adopts in the step (2) is the phenyl ureas basic element of cell division.
3, method as claimed in claim 2 is characterized in that: at first with indoles or naphthalene class growth hormone solution soaking test-tube plantlet base portion, and then test-tube plantlet changed on the 1/2MS solid culture medium that does not contain hormone cultivate in step (3).
4, method as claimed in claim 3 is characterized in that: the temperature of in the step (1) bud being carried out low temperature treatment is 9 ± 2 ℃, and the processing time is 1-5 days; The content of the phenyl ureas basic element of cell division in minimal medium is 0.5-2mg/L in the step (2); Growth hormone adopts heteroauxin, indolebutyric acid or methyl in the step (3), and growth hormone growth from solution cellulose content is 2mg/L-15mg/L, and the time that the test-tube plantlet base portion soaks is 1 day, and the continuation incubation time in solid culture medium is 5-15 days.
5, method as claimed in claim 4 is characterized in that: the time of in the step (1) bud being carried out low temperature treatment is 3 days; The phenyl ureas basic element of cell division adopts 2-chloro-4-pyridine radicals-N-phenylurea in the step (2), and its content in minimal medium is 1mg/L; Growth hormone growth from solution cellulose content is 5-10mg/L in the step (3).
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| CNB011298219A CN1133364C (en) | 2001-10-29 | 2001-10-29 | Fast excised African chrysanthemum propagation method |
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| CNB011298219A CN1133364C (en) | 2001-10-29 | 2001-10-29 | Fast excised African chrysanthemum propagation method |
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| CN1133364C CN1133364C (en) | 2004-01-07 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100420369C (en) * | 2006-04-30 | 2008-09-24 | 江汉大学 | Method for obtaining hormone-free tissue culture detoxified seedling of chrysanthemum for tea use |
| CN102415337A (en) * | 2011-09-13 | 2012-04-18 | 沈阳农业大学 | Breeding method of gerbera hybrida tissue culture plug seedlings |
| CN106069767A (en) * | 2016-06-29 | 2016-11-09 | 无锡南理工科技发展有限公司 | A kind of breeding method of African Chrysanthemum detoxification seedling |
| CN106105669A (en) * | 2016-06-29 | 2016-11-16 | 无锡南理工科技发展有限公司 | A kind of African Chrysanthemum transplants substrate and the application thereof of seedling exercising |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101411305B (en) * | 2008-11-25 | 2011-11-30 | 华南师范大学 | Method for quickly reproducing in-vitro pea nut |
-
2001
- 2001-10-29 CN CNB011298219A patent/CN1133364C/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100420369C (en) * | 2006-04-30 | 2008-09-24 | 江汉大学 | Method for obtaining hormone-free tissue culture detoxified seedling of chrysanthemum for tea use |
| CN102415337A (en) * | 2011-09-13 | 2012-04-18 | 沈阳农业大学 | Breeding method of gerbera hybrida tissue culture plug seedlings |
| CN106069767A (en) * | 2016-06-29 | 2016-11-09 | 无锡南理工科技发展有限公司 | A kind of breeding method of African Chrysanthemum detoxification seedling |
| CN106105669A (en) * | 2016-06-29 | 2016-11-16 | 无锡南理工科技发展有限公司 | A kind of African Chrysanthemum transplants substrate and the application thereof of seedling exercising |
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| CN1133364C (en) | 2004-01-07 |
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