Somatic embryogenesis and plant regeneration method of Rosa chinesis

A technology for embryogenesis and somatic embryos, applied in plant regeneration, botanical equipment and methods, horticultural methods, etc. The problem of low rate and regeneration rate can be achieved, and the effect of promoting the formation of cotyledons, promoting accumulation, and increasing the induction rate can be achieved.

Inactive Publication Date: 2013-01-23
HUNAN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The inventor has been engaged in the research on somatic embryogenesis and regeneration of Yueyuehong, and found that the organogenesis of Yueyuehong can be induced by hormone regulation, and buds can be directly produced, but the induction rate and regeneration rate are very low, and no similar zygotic embryos have been seen. somatic embryogenesis
Cannot form rapid, complete, juvenile and regenerated strains, and it is difficult to improve rose (Luna rose) through gene transformation

Method used

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  • Somatic embryogenesis and plant regeneration method of Rosa chinesis
  • Somatic embryogenesis and plant regeneration method of Rosa chinesis
  • Somatic embryogenesis and plant regeneration method of Rosa chinesis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Plant material

[0023] The annual semi-lignified branches of Yueyuehong (R. chinensis Jacq.) in the nursery of Beijing Forestry University were selected for surface disinfection (disinfected with 70% alcohol for 30 seconds, then placed in 0.3% sodium hypochlorite solution for 15 minutes, and finally with washed 3 times with sterile water). Placed in containing 6-BA 2.0mg / L, 0.1mg / L NAA and 3.0mg / L GA 3 The MS solid medium (the medium containing these hormones is named SP medium) was cultured for 2 months to produce sterile shoots. The sterile buds were subcultured to fresh SP medium every month, and all the medium was sterilized at 121°C and 125 kPa for 15 minutes after adjusting the pH value to 6.0. Culture conditions: the temperature is 23°C-27°C, 16 hours of light and 8 hours of darkness per day, and the light intensity is 1600Lux of ordinary white light.

[0024] The results showed that Yueyuehong grew normally and vigorously on SP medium, and the aver...

Embodiment 2

[0025] Induction of embodiment 2 callus

[0026] Put the aseptic buds with 1mm petiole on the EP medium (EP medium is the basic SH medium plus 3.0mg / L 2,4-D, 1mg / L TDZ and 300.0 mg / L L-preserved Amino acid, use 4.0g / L agarose to solidify the medium, adjust the pH value to 5.4), and culture in the dark at 23-27°C for 2 months.

[0027] The results showed that the medium containing 3mg / L 2,4-D and 1mg / LTDZ could produce light brown or transparent water-soaked embryogenic callus, and the induction rate was about 25%.

Embodiment 3

[0028] Example 3 Somatic Embryogenesis

[0029] Most of the embryogenic callus (usually brown loose particles, or transparent water-soaked callus) on the optimized EM medium (optimized EM medium is the basic SH medium plus 1.0mg / L 2, 4-D, 0.2 mg / L TDZ, 2.5 mg / L ABA, 3 mg / L GA 3 And 300.0 mg / L L-proline, use 4.0g / L agarose to solidify the medium. Adjust the pH value to 5.4) for culture, the culture temperature is 23°C-27°C, and the light is dark culture, or red light (400Lux, light 16h / dark 8h), or ordinary white light (1600Lux, light 16h / dark 8h). After two weeks spherical proembryos are produced (see figure 1 A). A part continued to differentiate into various types of embryos in subsequent cultures (see figure 1 B- figure 1 D).

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Abstract

The invention discloses a somatic embryogenesis and plant regeneration method of Rosa chinesis. The method is a process that sterile sprout tender leaves of Rosa chinesis (Rosa chinesis Jacq.) with leaves and stems are used as the raw materials and co-acted with SH culture mediums added with 2,4-D (2,4-dichlorophenoxyacetic acid), TDZ (thidiazuron) and L-proline so as to induce out calluses, and subsequently somatic embryos are induced under the action of red light so as to obtain regeneration plants. According to the method, somatic embryogenesis generated in a high frequency mode is obtained by using different culture mediums under conditions of dark cultivation, red light cultivation and ordinary white light respectively, and the regeneration rate or normal plants is more or less than 50%. The somatic embryogenesis and plant regeneration method in a high frequency generation has very wide use in aspects of gene transformation, gene function verification, intermediate propagation, tree-age rejuvenation, artificial seed creation, germplasm improvement and innovation.

Description

technical field [0001] The present invention relates to a method for somatic embryogenesis and plant regeneration of Chinese rose, especially a method for co-inducing somatic embryogenesis of Chinese rose with 2,4-D and TDZ, and the method rapidly develops in Chinese rose. Applications in reproduction, gene transformation, and artificial somatic embryos. Background technique [0002] Somatic embryogenesis (Somatic Embryogenesis) refers to the process in which plant somatic cells develop into new individuals through a similar pathway to zygotic embryos under in vitro culture conditions. Due to its characteristics of fast speed, large quantity, and complete structure, the use of somatic embryogenesis for artificial seedling cultivation of arbor species may become an important means for the source of plantation seedlings in the future; since somatic embryos are often produced from a single cell, through Gene transformation through the somatic embryogenesis pathway is conducive...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 陈己任熊兴耀邓子牛
Owner HUNAN AGRICULTURAL UNIV
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