CN104106467A - Tissue culture and rapid propagation method of sugarcane - Google Patents
Tissue culture and rapid propagation method of sugarcane Download PDFInfo
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- CN104106467A CN104106467A CN201410296653.9A CN201410296653A CN104106467A CN 104106467 A CN104106467 A CN 104106467A CN 201410296653 A CN201410296653 A CN 201410296653A CN 104106467 A CN104106467 A CN 104106467A
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Abstract
The invention discloses a tissue culture and rapid propagation method of sugarcane, and relates to the technical field of plant tissue culture. The method comprises the following steps of selecting sugarcane axillary buds; washing the sugarcane axillary buds clean with clear water; sterilizing; inoculating the sugarcane axillary buds to a bud-inducing culture medium; regulating light and temperature; culturing until new buds are grown; transferring the new buds to a seedling culture medium; regulating the light and temperature to obtain seedlings; transferring the seedlings to a rooting culture medium; regulating the light and temperature; culturing to obtain sugarcane tissue cultured seedlings; carrying out seedling-hardening culture; and transplanting the hardened seedlings to an open field for cultivation. The tissue culture and rapid propagation method shortens a propagation period of the sugarcane.
Description
Technical field
The present invention relates to field of plant tissue culture technique, especially a kind of sugarcane tissue-culture quick-breeding method.
Background technology
Sugarcane is grass family saccharum plant, is important in the world sugar [yielding.The cultivation of sugarcane, normally with the axillalry bud breeding on sugarcane stipes, utilizes the axillalry bud remaining on the stem foot in soil to do perennial root breeding.These method reproduction speeds are slow, gained cane seedling genetic character differs greatly, and the infecting of susceptible viral in cultivation process, cause its deterioration of variety serious.So, a kind of method of being badly in need of seeking high yield, high-quality, sugarcane breeding efficiently.
Summary of the invention
The object of this invention is to provide a kind of sugarcane tissue-culture quick-breeding method, this method can solve the slow problem of sugarcane reproduction speed.
In order to address the above problem, the technical solution used in the present invention is: a kind of sugarcane tissue-culture quick-breeding method, is characterized in that comprising the following steps:
A, get sugarcane axillalry bud, clear water is cleaned, the alcohol-pickled 45s～60s with 75%, then proceed to 0.05%～0.1% mercuric chloride solution and soak 20 minutes～30 minutes, sterile distilled water flushing 3 times～5 times;
B, be seeded to bud inducing culture, regulating illumination 1800LUX～2200LUX, temperature is 23 ℃～26 ℃, cultivates 10 days～15 days to growing sprouting; Wherein, described bud inducing culture, for take MS medium as minimal medium, adds 30 grams per liter～50 grams per liter sucrose and 10 grams per liter～15 grams per liter agar;
C, treat that sprouting grows to 2 centimetres～3 centimetres, proceed to strong seedling culture base, regulating illumination 2000LUX～2500LUX, temperature is 23 ℃～25 ℃, cultivates 15 days～18 days, to 1 centimetre～2 centimetres of heights of seedling; Wherein, described strong seedling culture base be take 1/2MS medium as minimal medium, adds 8 grams per liter～10 grams per liter carragheens, 0.5 grams per liter～1 grams per liter peptone;
D, transferred into root media above-mentioned strong sprout, regulating illumination 2200 Lux～2300 Lux, temperature is 23 ℃～26 ℃, cultivates and to seedling base portion, grows the root of 1 centimetre～3 centimetres in 12 days～15 days, obtains sugar-cane tissue culture seedlings; The described foster base of taking root be take 1/2MS medium as minimal medium, adds 1～1.5 grams per liter methyl α-naphthyl acetate, 0.1～0.2 grams per liter gibberellin and 2.5～3 grams per liter paclobutrazols;
E, hardening cultivate 25 days～30 days, transplant to open country and cultivate.
Owing to having adopted technique scheme, the present invention compared with prior art has following beneficial effect:
The present invention be take sugarcane axillalry bud and is organized cultivation as material, has greatly shortened the breeding cycle of sugarcane, and gained seedling early growth is healthy and strong, and genetic character is more consistent, and the breeding of sugarcane is not subject to seasonal restrictions, can spread.
Embodiment
Below in conjunction with embodiment, the invention will be further described:
Embodiment 1
Get sugarcane axillalry bud, with clear water, clean, the alcohol-pickled 45s with 75%, then proceed to 0.05% mercuric chloride solution immersion 30 minutes, sterile distilled water rinses 3 times; Be seeded to bud inducing culture, regulating illumination 1800LUX, regulating temperature is 23 ℃, cultivates 15 days; When sprouting grows to 2 centimetres, proceed to strong seedling culture base, regulating illumination 2000LUX, temperature is 23 ℃, cultivate 18 days, during to 1 centimetre of height of seedling, transfer into root media, regulating illumination 2200 Lux, temperature is 26 ℃, cultivates and to seedling base portion, grows the root of 3 centimetres in 12 days, obtains sugar-cane tissue culture seedlings; Hardening is cultivated 25 days, finally transplants to open country and cultivates.
Bud inducing culture in the present embodiment, for take MS medium as minimal medium, adds 30 grams per liter sucrose and 10 grams per liter agar;
Strong seedling culture base in the present embodiment be take 1/2MS medium as minimal medium, adds 8 grams per liter carragheens and 1 grams per liter peptone;
Taking root foster base in the present embodiment be take 1/2MS medium as minimal medium, adds 1 grams per liter methyl α-naphthyl acetate, 0.2 grams per liter gibberellin and 2.5 grams per liter paclobutrazols.
Embodiment 2
Get sugarcane axillalry bud, with clear water, clean, the alcohol-pickled 55s with 75%, then proceed to 0.1% mercuric chloride solution immersion 20 minutes, sterile distilled water rinses 4 times; Be seeded to bud inducing culture, regulating illumination 2000LUX, regulating temperature is 26 ℃, cultivates 13 days; When sprouting grows to 3 centimetres, proceed to strong seedling culture base, regulating illumination 2500LUX, temperature is 25 ℃, cultivate 15 days, during to 2 centimetres of heights of seedling, transfer into root media, regulating illumination 2300 Lux, temperature is 23 ℃, cultivates and to seedling base portion, grows the root of 1 centimetre in 12 days, obtains sugar-cane tissue culture seedlings; Hardening is cultivated 30 days, finally transplants to open country and cultivates.
Bud inducing culture in the present embodiment, for take MS medium as minimal medium, adds 50 grams per liter sucrose and 15 grams per liter agar;
Strong seedling culture base in the present embodiment be take 1/2MS medium as minimal medium, adds 10 grams per liter carragheens and 0.5 grams per liter peptone;
Taking root foster base in the present embodiment be take 1/2MS medium as minimal medium, adds 1.2 grams per liter methyl α-naphthyl acetates, 0.1 grams per liter gibberellin and 3 grams per liter paclobutrazols.
Embodiment 3
Get sugarcane axillalry bud, with clear water, clean, the alcohol-pickled 60s with 75%, then proceed to 0.1% mercuric chloride solution immersion 25 minutes, sterile distilled water rinses 5 times; Be seeded to bud inducing culture, regulating illumination 2200LUX, regulating temperature is 26 ℃, cultivates 10 days; When sprouting grows to 3 centimetres, proceed to strong seedling culture base, regulating illumination 2300LUX, temperature is 24 ℃, cultivate 17 days, during to 2 centimetres of heights of seedling, transfer into root media, regulating illumination 2300 Lux, temperature is 23 ℃, cultivates and to seedling base portion, grows the root of 1 centimetre in 12 days, obtains sugar-cane tissue culture seedlings; Hardening is cultivated 28 days, finally transplants to open country and cultivates.
Bud inducing culture in the present embodiment, for take MS medium as minimal medium, adds 50 grams per liter sucrose and 13 grams per liter agar;
Strong seedling culture base in the present embodiment be take 1/2MS medium as minimal medium, adds 9 grams per liter carragheens and 0.75 grams per liter peptone;
Taking root foster base in the present embodiment be take 1/2MS medium as minimal medium, adds 1.5 grams per liter methyl α-naphthyl acetates, 0.15 grams per liter gibberellin and 3 grams per liter paclobutrazols.
Claims (1)
1. a sugarcane tissue-culture quick-breeding method, is characterized in that comprising the following steps:
A, get sugarcane axillalry bud, clear water is cleaned, the alcohol-pickled 45s～60s with 75%, then proceed to 0.05%～0.1% mercuric chloride solution and soak 20 minutes～30 minutes, sterile distilled water flushing 3 times～5 times;
B, be seeded to bud inducing culture, regulating illumination 1800LUX～2200LUX, temperature is 23 ℃～26 ℃, cultivates 10 days～15 days to growing sprouting; Wherein, described bud inducing culture, for take MS medium as minimal medium, adds 30 grams per liter～50 grams per liter sucrose and 10 grams per liter～15 grams per liter agar;
C, treat that sprouting grows to 2 centimetres～3 centimetres, proceed to strong seedling culture base, regulating illumination 2000LUX～2500LUX, temperature is 23 ℃～25 ℃, cultivates 15 days～18 days, to 1 centimetre～2 centimetres of heights of seedling; Wherein, described strong seedling culture base be take 1/2MS medium as minimal medium, adds 8 grams per liter～10 grams per liter carragheens, 0.5 grams per liter～1 grams per liter peptone;
D, transferred into root media above-mentioned strong sprout, regulating illumination 2200 Lux～2300 Lux, temperature is 23 ℃～26 ℃, cultivates and to seedling base portion, grows the root of 1 centimetre～3 centimetres in 12 days～15 days, obtains sugar-cane tissue culture seedlings; The described foster base of taking root be take 1/2MS medium as minimal medium, adds 1～1.5 grams per liter methyl α-naphthyl acetate, 0.1～0.2 grams per liter gibberellin and 2.5～3 grams per liter paclobutrazols;
E, hardening cultivate 25 days～30 days, transplant to open country and cultivate.
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| CN201410296653.9A CN104106467A (en) | 2014-06-27 | 2014-06-27 | Tissue culture and rapid propagation method of sugarcane |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104823848A (en) * | 2015-05-05 | 2015-08-12 | 广州甘蔗糖业研究所 | Root-inducing strong seedling medium for sugarcane and direct field planting method of sugarcane tissue culture bottle seedling |
| CN107094623A (en) * | 2017-04-19 | 2017-08-29 | 江苏农林职业技术学院 | A kind of propagation method of sugarcane tissue culture |
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| CN101711504A (en) * | 2009-12-14 | 2010-05-26 | 湖南农业大学 | Rapid propagation method of triarrhena sacchariflora |
| CN101720668A (en) * | 2009-09-18 | 2010-06-09 | 广西壮族自治区农业科学院 | Method for tissue culturing and quick propagation of sugarcane with intermittent immersion bioreactor |
| CN101803574A (en) * | 2010-04-23 | 2010-08-18 | 广州甘蔗糖业研究所 | Detoxication and tissue culture rapid propagation method of chewing cane axillary buds |
| CN101904262A (en) * | 2010-07-16 | 2010-12-08 | 广西壮族自治区甘蔗研究所 | Ex vitro rooting method of sugarcane test tube plantlets |
| CN102090340A (en) * | 2010-12-22 | 2011-06-15 | 广州甘蔗糖业研究所湛江甘蔗研究中心 | Method for rapidly breeding sugarcane stem tip by tissue culture and virus removal |
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2014
- 2014-06-27 CN CN201410296653.9A patent/CN104106467A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101720668A (en) * | 2009-09-18 | 2010-06-09 | 广西壮族自治区农业科学院 | Method for tissue culturing and quick propagation of sugarcane with intermittent immersion bioreactor |
| CN101711504A (en) * | 2009-12-14 | 2010-05-26 | 湖南农业大学 | Rapid propagation method of triarrhena sacchariflora |
| CN101803574A (en) * | 2010-04-23 | 2010-08-18 | 广州甘蔗糖业研究所 | Detoxication and tissue culture rapid propagation method of chewing cane axillary buds |
| CN101904262A (en) * | 2010-07-16 | 2010-12-08 | 广西壮族自治区甘蔗研究所 | Ex vitro rooting method of sugarcane test tube plantlets |
| CN102090340A (en) * | 2010-12-22 | 2011-06-15 | 广州甘蔗糖业研究所湛江甘蔗研究中心 | Method for rapidly breeding sugarcane stem tip by tissue culture and virus removal |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104823848A (en) * | 2015-05-05 | 2015-08-12 | 广州甘蔗糖业研究所 | Root-inducing strong seedling medium for sugarcane and direct field planting method of sugarcane tissue culture bottle seedling |
| CN107094623A (en) * | 2017-04-19 | 2017-08-29 | 江苏农林职业技术学院 | A kind of propagation method of sugarcane tissue culture |
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