Culture medium for improving protoplast regeneration frequency of lupinus albus

A protoplast and white lupine technology, applied in the field of modern agriculture and plant biotechnology research, can solve the problems of callus regeneration plants, unfavorable lupine biotechnology, poor regeneration ability, etc.

Inactive Publication Date: 2012-07-11
YANGZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Lupine is an important economic plant, but its regenerative ability is very poor, not only from callus to regenerated plant, but also from plant to root. It is currently recognized in the world as a plant that is difficult to carry out biotechnology research through tissue culture methods one
At present, there is only one report about lupine protoplast regenerated plants at home and abroad, and only one regenerated plant was obtained. Therefore, the development of its biotechnology is limited, including protoplast fusion, embryo culture and microspore culture, etc.; Facilitating lupine-related biotechnological research

Method used

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  • Culture medium for improving protoplast regeneration frequency of lupinus albus
  • Culture medium for improving protoplast regeneration frequency of lupinus albus
  • Culture medium for improving protoplast regeneration frequency of lupinus albus

Examples

Experimental program
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Embodiment 1

[0018] The screening process selects the cultivar white lupine (Lupinus albus) as the experimental material; by cultivating 4-5 weeks of sterile seedlings, the young leaves are taken; the protoplasts are separated by enzymatic digestion; Injured tissue; regenerated shoots were obtained by selecting different medium combinations. The specific operation is as follows:

[0019] (1) White lupine seeds ( figure 1 A) Treat with 96% alcohol for 15 seconds, then treat with 3% sodium hypochlorite for 15 minutes, rinse with sterilized double distilled water for 3 times, then soak in sterilized double distilled water for 2 hours, so that the seeds can absorb water and germinate.

[0020] (2) Sterilized seeds ( figure 1 B) Put in 1 / 2 MS medium (without organic ingredients) + 20g sucrose, pH5.8, grow in light incubator for 4 weeks (25°C, 16h light / 8 hours dark, 1500lx).

[0021] (3) Get sterile young leaves ( figure 1 C) After enzymatic hydrolysis (0.5% Isozyme R-10, 1% cellulase 'Onoz...

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Abstract

The invention relates to a culture medium for improving the protoplast regeneration frequency of lupinus albus, and the method comprises the following components: a minimal medium, hormone and other components, wherein the minimal medium is composed of mass and trace of B5 and organic MS; the hormone is combined as follows: 0.5 mg / L NAA (Naphthyl Acetic Acid) + 2.0 mg / L 6-BA (Butyl Acrylate) + 1.0 mg / L TDZ (Thidiazuron) + 0.5 mg / L gibberellins; and the other components are combined as follows: 5 g / L cane sugar + 126 mg / L fructose + 120 mg / L ribose + 120 mg / L xylose + 126 mg / L mannose + 137 mg / L cellobiose + 200 mg / L casein acid hydrolysate +5 g / L silver nitrate + 8 g / L agarose. The culture medium can be used for solving the bottleneck problem of growing a lupinus albus callus into a plant, and has great significance to developing biotechnology related to lupinus albus.

Description

technical field [0001] The invention relates to a culture medium for increasing the regeneration frequency of lupine protoplasts, which belongs to the field of modern agriculture and plant biotechnology research. Background technique [0002] Lupine is a herbaceous plant with extremely high application value. Its seed protein content is as high as 50%, and its oil content is 5%-20%. It can not only be used as food, but also as high-quality feed. At the same time, the flower shape of lupine is beautiful, and the flowering period is more than 2 months. It has single color and multiple colors. It is very decorative when planted indoors, in courtyards, flower beds and other places. At present, it is planted in many countries in Europe, Australia, Asia and Africa. The widely cultivated varieties mainly include white lupine (Lupinus albus), narrow-leaved lupine (L. angustifolius) and yellow lupine (L. luteus). my country's lupine is limited to sporadic cultivation attempts, mainl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 王幼平蒋金金王娟刘银杜坤
Owner YANGZHOU UNIV
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