A method for plant regeneration of pure solid cultured hybrid Liriodendron somatic embryo
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A technology for hybridizing Liriodendron tulipifera and Liriodendron tulipifera body, applied in plant regeneration, botanical equipment and methods, horticultural methods, etc., can solve the problems of many steps, long time, and insufficient convenience, and achieve simple operation and seedling resistance good effect
Active Publication Date: 2017-11-24
NANJING FORESTRY UNIV
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Problems solved by technology
This culture method has many steps, takes a long time, and is not convenient enough. It needs to be further innovated to meet the needs of use.
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Embodiment 1
[0029] A method for rapid propagation of hybrid Liriodendron somatic embryo solid culture, comprising the following steps:
[0030] 1) Solid culture of embryogenic callus of Liriodendron chinensis
[0031] 3 / 4MS+VC5mg / L+sucrose 30g / L was used as the basic medium, and different concentrations of amino-oligosaccharins were added to form the somatic embryo induction medium of Liriodendron tulipifera. The basic medium without amino oligosaccharin was used as the control. Four genotypes were used to hybridize the embryogenic callus of Liriodendron tulipifera, and nine concentration gradients were set up (A1 0mg / L, A2 0.005mg / L, A3 0.01mg / L, A40.05mg / L, A5 0.1mg / L, A6 1mg / L, A7 10mg / L, A8 100mg / L, A9 500mg / L), at least 3 dishes were inoculated for each concentration, and 5 pieces of callus were connected to each dish. The diameter of each piece of callus was about 0.5cm, and the weight was about 1.6g / dish. The initial culture environment was dark culture at 24°C.
[0032] will b...
Embodiment 2
[0057] A method for rapid propagation of hybrid Liriodendron somatic embryo solid culture, with embodiment 1, wherein in the amino-oligosaccharin culture medium of optimal concentration (3 / 4MS+VC5mg / L+amino-oligosaccharin 0.01~0.1mg / L+sucrose 30g / L) was added with 2.0mg / L ABA, and a new round of somatic embryo induction culture was carried out for genotypes 253010, 112040, and C136. The experiment was repeated 3 times, and the state of somatic embryogenesis was observed and recorded every 5 days. When somatic embryos matured, the number of somatic embryos was counted by weighing method. Statistical analysis was performed on genotype 253010 cultured for 56 days, and genotypes C136, 82133, and 112040 cultured for 84 days, and each subculture time was 28 days.
[0058] Select amino-oligosaccharin 0.01mg / L as the culture concentration, add 2.0mg / L ABA, that is, induce somatic embryo medium formula: 3 / 4MS+VC5mg / L+amino-oligosaccharin 0.01mg / L+ABA2.0mg / L+ Sucrose 30g / L, pH=5.72, so...
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Abstract
The invention discloses a regeneration method adopting pure solid culture for a somatic embryo plant of liriodendron chinense. The method comprises steps as follows: 1) hybrid liriodendron chinense embryonic calli are induced; 2) 3 / 4 MS, VC with concentration of 5mg / L, saccharose with concentration of 30g / L and amino oligochitosan with concentration of 0.01-0.1mg / L are taken as basic media, and the hybrid liriodendron chinense embryonic calli are cultured in dark at 24 DEG C; 3) the material is transferred to a tissue-culture vessel in a light culture room and cultured continuously with the 3 / 4 MS basic medium until the regeneration plant is formed; 4) the regeneration plant is acclimated and transplanted. According to the regeneration method adopting pure solid culture for the somatic embryo plant of liriodendron chinense, the pure solid culture manner is adopted, the somatic embryo induction links including suspension culture and plate culture in the somatic embryo generation process are skipped, a hybrid liriodendron chinense somatic embryo generation system is optimized and enriched, and accordingly, the high-quality somatic embryo regeneration plant is obtained more efficiently and more conveniently; a stable and high-frequency hybrid liriodendron chinense somatic embryo generation system adopting pure solid media and having properties of being easy to operate and good in seedling resistance is established, and reference is provided for establishment of a more efficient genetic transformation receptor system of hybrid liriodendron chinense.
Description
technical field [0001] The invention belongs to the technical field of plant breeding, and in particular relates to a plant regeneration method for pure solid culture of somatic embryos of Liriodendron chinensis. Background technique [0002] The tissue culture of Liriodendron began in 1986. American Sommer et al. conducted tissue culture experiments on Liriodendron tulipifera and obtained some plants. Parks et al. used immature embryos to cultivate Liriodendron tulipifolia. The most notable achievement in domestic research on somatic embryos of Liriodendron chinensis is that Chen Jinhui and others established a complete somatic embryogenesis system of Liriodendron chinensis by using immature embryos as explants, through callus induction and suspension culture , and somatic embryo rapid seedling formation system. In this study, the best culture schemes were optimized for each culture stage of somatic embryogenesis of Liriodendron chinensis, including callus induction, subc...
Claims
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