CN101642049A - Plant regrowing method for western balsam poplar isolated organ - Google Patents
Plant regrowing method for western balsam poplar isolated organ Download PDFInfo
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- CN101642049A CN101642049A CN200910035029A CN200910035029A CN101642049A CN 101642049 A CN101642049 A CN 101642049A CN 200910035029 A CN200910035029 A CN 200910035029A CN 200910035029 A CN200910035029 A CN 200910035029A CN 101642049 A CN101642049 A CN 101642049A
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Abstract
The invention discloses a plant regrowing method for a western balsam poplar isolated organ, comprising the steps: cultivating the stem segment or the leafstalk of a western balsam poplar in an MS culture medium containing 0.001 to 0.005 mg/L of TDZ for 9 to 10 days; grafting to an MS culture medium containing 0.001 to 0.002 mg/L of TDZ for 14 to 16 days; inducing adventitious buds; continuously grafting the induced adventitious buds to the MS culture medium containing 0.001 to 0.002 mg/L of TDZ for strengthening seedlings; reaching the height of the seedlings of 4 to 5 cm after 18 to 22 days;grafting to an MS culture medium with a large amount of 1/2 elements and growing roots directly after 18 to 22 days. The plant regrowing method for a western balsam poplar isolated organ is easy to operate and realize and creates basic conditions for researching on poplar functional genes.
Description
Technical field
The invention belongs to plant tissue culture regeneration techniques field, be specifically related to the method for comospore poplar isolated organ regeneration plant.
Background technology
The comospore poplar mainly is distributed in Western North America, belongs to more unobtainable relevant material in China.USDOE's Oak Ridge National Laboratory (ORNL) in 2002 and associating Joint Genome Institute (JGI) have started the complete genomic order-checking plan of willow, and its material source is in the clone (Nisqully-1) of comospore poplar (Populus trichocarpa).The inventor furthers investigate this clonal tissue culture, has set up rapid propagation system.
Summary of the invention
Technical problem to be solved by this invention provides a kind of method of comospore poplar isolated organ regeneration plant, with fill up domestic so that the world blank.
For solving the problems of the technologies described above, the technical solution used in the present invention is as follows:
The method of comospore poplar isolated organ regeneration plant, the stem section of comospore poplar or petiole were cultivated 9～10 days containing on the MS medium of 0.001～0.005mg/LTDZ, be forwarded to again on the MS medium that contains 0.001～0.002mg/L TDZ and cultivated 14～16 days, induce indefinite bud; The indefinite bud that induces is continued subculture carry out strong sprout containing on the MS medium of 0.001～0.002mg/L TDZ, after 18～22 days, height of seedling can reach 4～5cm, transfers on macroelement is 1/2 MS medium again, can directly take root in 18～22 days.
Wherein, optimized technical scheme is: the stem section of comospore poplar or petiole were cultivated 9～10 days containing on the MS medium of 0.003～0.005mg/L TDZ, be forwarded on the MS medium that contains 0.001mg/L TDZ again and cultivated 14～16 days, induce indefinite bud.Preferred technical scheme is: the stem section of comospore poplar or petiole were cultivated 9～10 days containing on the MS medium of 0.005mg/L TDZ, be forwarded on the MS medium that contains 0.001mg/L TDZ again and cultivated 14～16 days, induce indefinite bud.Most preferred technical scheme is: the stem section of comospore poplar or petiole were cultivated 10 days containing on the MS medium of 0.005mg/L TDZ, be forwarded on the MS medium that contains 0.001mg/L TDZ again and cultivated 15 days, induce indefinite bud.
Wherein, optimized technical scheme is: the indefinite bud that induces is continued subculture carry out strong sprout containing on the MS medium of 0.001mg/L TDZ, after 18～22 days, height of seedling can reach 4～5cm, transfer again on macroelement is 1/2 MS medium, can directly take root in 18～22 days.Most preferred technical scheme is: the indefinite bud that induces is continued subculture carry out strong sprout containing on the MS medium of 0.001mg/L TDZ, after 20 days, height of seedling can reach 4～5cm, transfers on macroelement is 1/2 MS medium again, can directly take root in 20 days.
Beneficial effect: the method for comospore poplar isolated organ regeneration plant of the present invention, easy operating and realization have been created primary condition for carrying out the research of willow functional gene.
Embodiment:
According to following embodiment, the present invention may be better understood.Yet, those skilled in the art will readily understand that the described concrete material proportion of embodiment, condition of culture and result thereof only are used to illustrate the present invention, and should also can not limit the present invention described in detail in claims.
Embodiment 1: the different basic elements of cell division are to the influence of adventitious bud inducing.
1 materials and methods
1.1 material:
Comospore poplar test-tube plantlet.
1.2 method:
Adopt stem section, petiole and the blade evoking adventive bud of aseptic seedling respectively, with MS is minimal medium, 6-BA (six-benzyl purine), TDZ (Thidiazuron), 2ip (isopentennyladenine) and ZT (zeatin) exogenous hormone of additional variable concentrations, sucrose concentration is 3% (w/w), pH5.7, intensity of illumination 2000Lx, illumination every day 16h, 23～28 ℃ of temperature.
2 results and analysis:
Long blade for stem section, petiole and 1 * 1cm size about 1cm is inoculated in respectively on the medium of the additional various basic elements of cell division of MS, the results are shown in Table 1 behind the cultivation 20d.
As shown in Table 1, three kinds of explants all can't directly induce indefinite bud on the medium of additional 4 kinds of variety classes basic elements of cell division respectively at MS.Stem section and petiole can induce callus, and inductivity does not wait 0～100%, and the blade inductivity is zero.The increase stem section callus induction rate of TDZ concentration containing on the medium of TDZ, along with can reach 100%.The callus that the medium of different exogenous hormone combinations produces all has vitrification phenomenon in various degree, and serious to some extent with the cytokinin concentration increase.
The different basic elements of cell division of table 1 are to the influence of adventitious bud inducing
(continuous table 1)
Annotate: A is the callus that stem section end produces, and the callus that B produces for the petiole end, C are the callus (nothing) that blade produces
Embodiment 2: best group is trained groping of regeneration condition.
In view of the MS medium callus induction rate of additional TDZ the highest, so its further test-induced indefinite bud of selection.In this test because of the easy evoked callus of high concentration of cytokinin also easily produces vitrification phenomenon simultaneously, so employing high-concentration and low-concentration TDZ cultivates in succession.Cultivate about 10d containing on the MS medium of higher concentration TDZ, when the explant end occur expanding but before not producing vitrifying as yet subculture on the MS of TDZ low concentration medium, can produce a large amount of indefinite buds at the explant base portion that expands about 15d, growth coefficient is 8～10 times (the results are shown in Table 2).
Can see by table 2 and to know, explant is transformed into MS+TDZ0.001mg/L or MS+TDZ0.002mg/L after MS+TDZ0.001～0.005mg/L cultivates 10d, adventitious bud induction frequency improves gradually, cultivate 10d by MS+TDZ0.005～0.5mg/L and be transformed into MS+TDZ0.001mg/L or MS+TDZ0.002mg/L, indefinite bud increases with TDZ concentration and reduces, it is the highest wherein to be transformed into the MS+TDZ0.001mg/L adventitious bud induction frequency with MS+TDZ0.005mg/L, and does not have vitrification phenomenon.
The indefinite bud that induces is continued subculture carry out strong sprout on the MS+TDZ0.001mg/L medium, about 20d left and right sides height of seedling can reach 4～5cm..Transfer on macroelement is 1/2 MS medium, can directly take root about 20d.
Table 2 high-concentration and low-concentration TDZ cultivates the influence to adventitious bud inducing and vitrification phenomenon in succession
(continuous table 2)
Annotate: it is a small amount of below 20% that vitrification phenomenon accounts for inoculation several, and 40～60% is more, is many more than 70%.
Claims (6)
1, the method for comospore poplar isolated organ regeneration plant, it is characterized in that the stem section of comospore poplar or petiole were cultivated 9～10 days containing on the MS medium of 0.001～0.005mg/L TDZ, be forwarded to again on the MS medium that contains 0.001～0.002mg/L TDZ and cultivated 14～16 days, induce indefinite bud; The indefinite bud that induces is continued subculture carry out strong sprout containing on the MS medium of 0.001～0.002mg/L TDZ, after 18～22 days, height of seedling can reach 4～5cm, transfers on macroelement is 1/2 MS medium again, can directly take root in 18～22 days.
2, the method for comospore poplar isolated organ regeneration plant according to claim 1, it is characterized in that the stem section of comospore poplar or petiole were cultivated 9～10 days containing on the MS medium of 0.003～0.005mg/L TDZ, be forwarded to again on the MS medium that contains 0.001mg/L TDZ and cultivated 14～16 days, induce indefinite bud.
3, the method for comospore poplar isolated organ regeneration plant according to claim 2, it is characterized in that the stem section of comospore poplar or petiole were cultivated 9～10 days containing on the MS medium of 0.005mg/L TDZ, be forwarded to again on the MS medium that contains 0.001mg/LTDZ and cultivated 14～16 days, induce indefinite bud.
4, the method for comospore poplar isolated organ regeneration plant according to claim 3, it is characterized in that the stem section of comospore poplar or petiole were cultivated 10 days containing on the MS medium of 0.005mg/L TDZ, be forwarded to again on the MS medium that contains 0.001mg/LTDZ and cultivated 15 days, induce indefinite bud.
5, the method for comospore poplar isolated organ regeneration plant according to claim 1, it is characterized in that will induce indefinite bud continue subculture and carry out strong sprout containing on the MS medium of 0.001mg/L TDZ, after 18～22 days, height of seedling can reach 4～5cm, transfer again on macroelement is 1/2 MS medium, can directly take root in 18～22 days.
6, the method for comospore poplar isolated organ regeneration plant according to claim 5, it is characterized in that will induce indefinite bud continue subculture and carry out strong sprout containing on the MS medium of 0.001mg/L TDZ, after 20 days, height of seedling can reach 4～5cm, transfer again on macroelement is 1/2 MS medium, can directly take root in 20 days.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103461123A (en) * | 2013-09-13 | 2013-12-25 | 东北林业大学 | Method for inducing regeneration of adventitious bud of western balsam poplar by histone deacetylase inhibitor |
| CN104396754A (en) * | 2014-11-25 | 2015-03-11 | 东北林业大学 | Populus trichocarpa Torr.Gray leaf adventitious bud induction and plant regeneration method |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103202231B (en) * | 2013-04-25 | 2016-03-16 | 陕西理工学院 | The method for building up of tissue culture system of gray poplar |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101341853B (en) * | 2008-08-04 | 2011-07-06 | 湖南茂源林业有限责任公司 | Tissue culture method for Beijing poplar |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103461123A (en) * | 2013-09-13 | 2013-12-25 | 东北林业大学 | Method for inducing regeneration of adventitious bud of western balsam poplar by histone deacetylase inhibitor |
| CN103461123B (en) * | 2013-09-13 | 2015-06-10 | 东北林业大学 | Method for inducing regeneration of adventitious bud of western balsam poplar by histone deacetylase inhibitor |
| CN104396754A (en) * | 2014-11-25 | 2015-03-11 | 东北林业大学 | Populus trichocarpa Torr.Gray leaf adventitious bud induction and plant regeneration method |
| CN104396754B (en) * | 2014-11-25 | 2016-08-31 | 东北林业大学 | A kind of comospore Folium Populi Pseudo-simonii sheet Induce aerosor and the method for plant regeneration |
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