CN102823493B - Method of sugarcane distant hybrid embryo in-vitro culture and seedling breeding - Google Patents
Method of sugarcane distant hybrid embryo in-vitro culture and seedling breeding Download PDFInfo
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- CN102823493B CN102823493B CN 201210296251 CN201210296251A CN102823493B CN 102823493 B CN102823493 B CN 102823493B CN 201210296251 CN201210296251 CN 201210296251 CN 201210296251 A CN201210296251 A CN 201210296251A CN 102823493 B CN102823493 B CN 102823493B
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Abstract
The invention relates to a method of sugarcane distant hybrid embryo in-vitro culture seedling growing, and belongs to the field of sugarcane genetic breeding research; the invention adopts sugarcane hybrid embryos as explants; the sugarcane distant hybrid embryos are inoculated on a modified KC germination medium for germination culture; after seedling emergence, the seedlings are transferred to a modified N6 rooting medium for root induction; the sugarcane hybrid embryo explants are sugarcane hybrid spica seeds after artificial peeling; the invention simplifies the seedling growing program, shortens the seedling growing period, reduces labor force cost, also improves the germination rate and seedling rate of sugarcane hybrid seeds, and provides reference for seedling growing in related sugarcane scientific research units and breeding facilities.
Description
Technical field
The present invention relates to the method that a kind of sugarcane distant hybrid embryo cultured in vitro is grown seedlings, belong to sugarcane genetic breeding research field.
Background technology
Distant hybridization is the important channel of carrying out Plant genetics, thremmatology and Study on Evolution, has now become one of key link that plant breeding makes a breakthrough.The cultured in vitro technology is a simple and practical biotechnology, and the equipment of requirement is simple, just can carry out in common Plant Tissue Breeding laboratory.This technology is applied to save abortion or hypogenetic distant hybrid embryo, hybrid seed is carried out culture in vitro, thereby can improve the seed planting percent, the seedling offspring that acquisition can be educated.At present, embryo cultured in vitro technology has been widely used in cereal crops such as paddy rice, wheat, and the application prospect in economic crops such as sugarcane also is worth expectation very much.But, because it is very complicated to influence the factor of hybrid embryonic development, and the condition of different plant species Different Cross Combinations hybrid embryonic development has nothing in common with each other, cause the cultured in vitro rescue of hybrid embryo to be difficult to sequencing, must carry out the optimization of various influence factors at different hybrid embryos, set up the hybrid embryo cultured in vitro that is applicable to different plant species and the combination system of growing seedlings.
Summary of the invention
The method that the object of the present invention is to provide a kind of sugarcane distant hybrid embryo cultured in vitro to grow seedlings, be that a kind of energy simple and effective ground passes through the effective ways that sugarcane hybrid seed cultured in vitro obtains the seedling offspring, this method is conducive to improve germination rate and the planting percent of sugarcane hybrid seed.
The present invention is achieved through the following technical solutions: the method that a kind of sugarcane distant hybrid embryo cultured in vitro is grown seedlings, it is characterized in that with sugarcane hybrid embryo as explant, be inoculated into earlier on the KC germination medium of improvement and sprout cultivation, transfer to root induction on the N6 root media of improvement after emerging;
Described sugarcane hybrid embryo explants is the sugarcane hybridization flower fringe seed behind the manual removal;
The KC germination medium prescription of described improvement is: KC+1 mg/L IBA+30g/L sucrose+4g/L agar, pH5.8;
The N6 culture of rootage based formulas of described improvement is: N6(macroelement, other components unchanged)+2 mg/L NAA+30g/L sucrose+4g/L agar+0.2g/L active carbon, pH5.8.
The present invention spends the fringe seed as culture materials with the sugarcane distant hybridization of various combination, tested MS, Knudson C(KC), multiple improved culture medium such as N6, and the medium that adds different 2.4-D and IBA concentration compared research, finally obtained germination medium and root media that suitable sugarcane distant hybrid embryo cultured in vitro is used, set up one and overlap the method that the embryo of sugarcane distant hybrid easily cultured in vitro is grown seedlings.
The present invention with the sugarcane hybrid embryo of Different Cross Combinations as explant, be inoculated into earlier on the KC germination medium of improvement and sprout cultivation, transfer to root induction on the N6 root media of improvement after emerging, thick shape body around behind the attention seed germination should cut, be conducive to surviving of seedling in the seedling culture of rootage, this method can make sugarcane hybrid embryo germination rate and the planting percent of taking root be significantly improved.
Described sugarcane hybrid embryo explants is the sugarcane hybridization flower fringe seed behind the manual removal, handles through 70% alcohol disinfecting 60s and 1 ‰ mercuric chloride solutions sterilization 6min successively.
The KC germination medium prescription of described improvement is: KC+1 mg/L IBA+30g/L sucrose+4g/L agar, and pH 5.8; The N6 culture of rootage based formulas of improvement is: N6(macroelement, other components unchanged)+2 mg/L NAA+30g/L sucrose+4g/L agar+0.2g/L active carbon, pH5.8.
The sugarcane distant hybrid embryo cultured in vitro seedling-cultivating method that the present invention proposes is compared with conventional culturing sugarcane seedlings technology has remarkable advantage, owing to adopted germination medium and root media through the test improvement to carry out the stage cultivation, make sugarcane hybrid blastocyte emerge and the differentiation of the cell of the different phase of taking root has obtained abundant propagation, only used emerging about 10 days to cultivate and culture of rootage cycle in 2 weeks the seedling that can obtain taking root.The present invention has simplified the program of growing seedlings, and has shortened growing-seedling period, has reduced labor cost, has improved the germination rate of sugarcane distant hybrid embryo and the planting percent of taking root, and can be relevant sugarcane R﹠D institution and breeding agency and is carrying out providing reference to use for reference when seedling is grown seedlings.
Embodiment:
Sugarcane distant hybrid embryo cultured in vitro seedling-cultivating method, the instrument and equipment of using in concrete the enforcement has: superclean bench, autoclave, low temperature refrigerator and illumination box etc., its detailed implementation step is as follows:
1. hybridization flower fringe seed is peeled off
Gather sugarcane hybridization flower fringe, manual removal is left and taken hybrid seed, and it is standby to be saved to-20 ℃ of low temperature refrigerators.
2. medium preparation
According to following culture medium prescription, prepare KC/N6 medium mother liquor successively and preserve standby in 4 ℃ of refrigerators of ﹣.
Induce seed germination KC improved culture medium: KC+1 mg/L IBA+30g/L sucrose+4g/L agar;
Induce the seedling N6 improved culture medium of taking root: N6(macroelement, other components unchanged)+2 mg/L NAA+30g/L sucrose+4g/L agar+0.2g/L active carbon;
Above medium is solid culture medium, pH5.8.
3. seed disinfection inoculation and cultured in vitro
The sugarcane seed that strips is wrapped with sterile gauze and is placed on aseptic bubbly water and washes 3 times, the alcohol disinfecting 60s of 70% water content, and 1 ‰ mercuric chloride solutions sterilization 6min, aseptic bubble is washed 5 times; With the spoon of the bacterium of going out seed is got on the filter paper to inhale during inoculation and remove surface moisture, seed is inoculated on the KC germination medium with tweezers in the other dry back of alcolhol burner, seal and place illumination box to cultivate to observe; When treating seed sprouting length to 4-5cm (about 10 days), cut the thick shape body of seedling root, be transferred to illumination incubator continuation cultivation on the N6 root media, seedling begins to take root after 1 week, can obtain all right group training seedling of root growth after 2 weeks.
Claims (2)
1. method that sugarcane distant hybrid embryo cultured in vitro is grown seedlings is characterized in that with sugarcane hybrid embryo as explant, is inoculated into earlier on the KC germination medium of improvement to sprout cultivation, transfers to root induction on the N6 root media of improvement after emerging;
Sugarcane hybrid embryo explants is the sugarcane hybridization flower fringe seed behind the manual removal;
The KC germination medium prescription of described improvement is: KC+1 mg/L IBA+30g/L sucrose+4g/L agar, and pH 5.8;
The N6 culture of rootage based formulas of described improvement is: N6+ 2 mg/L NAA+30g/L sucrose+4g/L agar+0.2g/L active carbon, pH5.8, described N6 refer to that macroelement reduces by half in the N6 medium, other components unchanged.
2. a kind of sugarcane distant hybrid embryo cultured in vitro according to claim 1 method of growing seedlings is characterized in that step is as follows:
The N6 culture of rootage based formulas of described improvement is: N6+ 2 mg/L NAA+30g/L sucrose+4g/L agar+0.2g/L active carbon, pH5.8, described N6 refer to that macroelement reduces by half in the N6 medium, other components unchanged.
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| CN 201210296251 CN102823493B (en) | 2012-08-20 | 2012-08-20 | Method of sugarcane distant hybrid embryo in-vitro culture and seedling breeding |
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| CN102823493B true CN102823493B (en) | 2013-07-17 |
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| CN103081606B (en) * | 2013-01-30 | 2014-05-07 | 云南省农业科学院甘蔗研究所 | Method for sugarcane spica seed sprouting and sprouted seedling temporary-planting |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101138321A (en) * | 2007-08-22 | 2008-03-12 | 浙江省农业科学院 | Sugarcane detoxication tissue culturing and fast propagating method |
| CN101401549A (en) * | 2008-11-04 | 2009-04-08 | 广州甘蔗糖业研究所 | Once-seedling forming quick propagating method for sugarcane tissue culture |
| CN101715730A (en) * | 2009-12-17 | 2010-06-02 | 昆明理工大学 | Tissue culture rapid propagation method for sugarcane new Taisugar No.10 |
| CN102118965A (en) * | 2008-07-24 | 2011-07-06 | 德克萨斯A&M大学体系 | Intergeneric hybrid plants between sorghum and saccharum and methods for production thereof |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101138321A (en) * | 2007-08-22 | 2008-03-12 | 浙江省农业科学院 | Sugarcane detoxication tissue culturing and fast propagating method |
| CN102118965A (en) * | 2008-07-24 | 2011-07-06 | 德克萨斯A&M大学体系 | Intergeneric hybrid plants between sorghum and saccharum and methods for production thereof |
| CN101401549A (en) * | 2008-11-04 | 2009-04-08 | 广州甘蔗糖业研究所 | Once-seedling forming quick propagating method for sugarcane tissue culture |
| CN101715730A (en) * | 2009-12-17 | 2010-06-02 | 昆明理工大学 | Tissue culture rapid propagation method for sugarcane new Taisugar No.10 |
Non-Patent Citations (6)
| Title |
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| "Plant Regeneration Via Somatic Embryogenesis in Sugarcane";B. S. Ahloowalia等;《Plant Cell Reports》;19831231;第2卷;第21-25页 * |
| "新台糖16号甘蔗组织培养技术研究";徐新江;《现代农业科技》;20101231(第21期);第50-51页 * |
| "甘蔗组织培养研究进展";杨柳等;《安徽农业科学》;20071231;第35卷(第12期);第3490-3492页 * |
| B. S. Ahloowalia等."Plant Regeneration Via Somatic Embryogenesis in Sugarcane".《Plant Cell Reports》.1983,第2卷第21-25页. |
| 徐新江."新台糖16号甘蔗组织培养技术研究".《现代农业科技》.2010,(第21期),第50-51页. |
| 杨柳等."甘蔗组织培养研究进展".《安徽农业科学》.2007,第35卷(第12期),第3490-3492页. |
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