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Somatic embryo all-solid cultivation plant regeneration method of hybridized liriodendron tulipifera by deionized formamide

A technology of deionized formamide and hybrid Liriodendron, which is applied in plant regeneration, botany equipment and methods, horticultural methods, etc., can solve the problems of inconvenient, long time, and many steps, and achieve good seedling resistance and easy operation. Simple, powerful effect of somatic embryo induction

Active Publication Date: 2016-12-14
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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.

Method used

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  • Somatic embryo all-solid cultivation plant regeneration method of hybridized liriodendron tulipifera by deionized formamide
  • Somatic embryo all-solid cultivation plant regeneration method of hybridized liriodendron tulipifera by deionized formamide
  • Somatic embryo all-solid cultivation plant regeneration method of hybridized liriodendron tulipifera by deionized formamide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] A method for rapid propagation of hybrid Liriodendron somatic embryo solid culture, comprising the following steps:

[0031] 1) Solid culture of embryogenic callus of Liriodendron chinensis

[0032] 3 / 4MS+VC5mg / L+sucrose 30g / L was used as the basic medium, and different concentrations of liquid deionized formamide (Formamide Deionized) were added, with a purity of >99.5%, to form a somatic embryo induction medium for hybrid Liriodendron chinensis. The basic medium without deionized formamide was used as the control. Using 3 genotypes and 7 concentration gradients (F1 0mg / L, F2 0.05mg / L, F3 0.1mg / L, F4 0.5mg / L, F5 1mg / L, F6 5mg / L, F7 10mg / L), At least 3 dishes were inoculated for each concentration, and 5 pieces of callus were connected to each dish. The diameter of each callus was about 0.5 cm, and the mass was about 1.6 g / dish. The initial culture environment was dark culture at 24°C. Each subculture time is 28d.

[0033] right figure 1 The somatic embryos of vari...

Embodiment 2

[0051] A method for rapid propagation of hybrid Liriodendron somatic embryo solid culture, with embodiment 1, wherein in the optimal concentration of deionized formamide medium (3 / 4MS+VC5mg / L+deionized formamide 0.1mg / L+sucrose 30g / L) was added with 2.0 mg / L ABA, and a new round of somatic embryo induction culture was carried out for genotypes 253010, 112040, and C136. The experiment was repeated three times, and the state of somatic embryogenesis was observed and recorded every 5 days. When mature, the number of somatic embryos was counted by weighing method. The results are shown in Table 2.

[0052] Table 2 The effect of deionized formamide combined with ABA on the induction of hybrid Liriodendron somatic embryos

[0053]

[0054]

[0055] Note: the material used in this example is a new material, so it is slightly different from the data in Example 1.

[0056] Table 2 shows that the relationship between the somatic embryogenesis speed of Liriodendron chinensis is ...

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Abstract

The invention discloses a somatic embryo all-solid cultivation plant regeneration method of hybridized liriodendron tulipifera by deionized formamide. The method includes the steps of: 1) inducing embryogenic callus of hybridized liriodendron tulipifera; 2) dark-culturing the embryogenic callus of the hybridized liriodendron tulipifera at 24 DEG C with a basic culture medium of 3 / 4MS + 5 mg / L of VC + 30 g / L of saccharose + 0.01-1 mg / L of the deionized formamide; 3) moving the material to a tissue culture bottle in a light culturing room, and continuously culturing the material with 3 / 4MS as a basic culture medium to obtain a regenerated plant; and 4) domesticating and transplanting the regenerated plant. Through the all-solid cultivation method, two somatic embryo, suspension cultivation and plate cultivation, during a somatic embryo generation process are omitted, thereby optimizing and enriching the somatic embryo regeneration system of the hybridized liriodendron tulipifera and further more high-effectively and conveniently obtaining high-quality somatic embryo regeneration regenerated plants. The method can establish the all-solid culture medium somatic embryo generation system, which is stable and high-frequent, is simple in operation and has excellent nursery stock resistance, of the hybridized liriodendron tulipifera, and provides references for establishing a more high-effective genetic transformation receptor system of the hybridized liriodendron tulipifera.

Description

technical field [0001] The invention belongs to the technical field of plant breeding, and in particular relates to a plant regeneration method for hybrid Liriodendron tulipifera somatic embryo pure solid culture by using deionized formamide. 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, in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 成铁龙陈金慧周敏敏施季森刘欣陆凤娟盛宇
Owner NANJING FORESTRY UNIV
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