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Immature-embryo culture method for red-peel peanuts

A technology for immature embryo culture and red-skinned peanuts, which is applied in the field of plant tissue culture, can solve the problems of difficulty in plant growth, difficulty in differentiation into plant growth, low regeneration frequency, etc., shorten the period required for tissue culture, and have significant social benefits and scientific research value Effect

Inactive Publication Date: 2017-02-15
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Peanut is a kind of large-grained leguminous plant. There have been reports on the successful in vitro regeneration of peanuts at home and abroad: McKently et al. used cotyledons with embryos and cotyledons without embryos as explants to induce clustered buds, and found that cotyledons with embryos were the most suitable explants. , grew into a complete plant after 258 days, but the process of obtaining explants in the above studies was complicated and required a large amount of seeds; Charleen and Venkatachala respectively used embryonic leaflets as explants, and successfully induced embryogenic callus through somatic embryo regeneration. tissue, but there is a problem of differentiation into plants; Mroginski et al. used immature leaflets of 3-5d seedling age as explants to induce clustered buds and obtained complete plants; Tiwari et al. used immature embryonic leaflets as explants to study the pre-culture time , medium and genotype on the regeneration of embryonic leaflets, the regeneration rate of the whole process can reach 80%, and it takes 4 months, but the above-mentioned regeneration system using embryonic leaflets as explants still has difficulty in plant formation and low repeatability At the same time, the problems of low regeneration frequency and long cycle have yet to be solved. At the same time, studies have shown that plant regeneration in vitro has a strong dependence on genotype

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] A method for culturing immature embryos of red-skinned peanuts, the technical scheme of which comprises the following steps:

[0027] (1) Remove the peel of the mature peanut pods, select the seeds with full particles and smooth surface, rinse with water, absorb the surface water, and soak in 75% ethanol solution with volume percentage for 1min, and 0.1% by mass mercuric chloride soak in the solution for 9 minutes, then rinse with sterile water for 4 times, peel off the seed coat of peanut seeds, soak in sterile water for 6 hours, and obtain sterile peanut embryo leaflets;

[0028] (2) Take the sterile peanut embryo leaflets obtained in step (1) and inoculate them on the cluster bud induction medium, the cluster bud induction medium is MS+0.1mg / L NAA+5mg / L6-BA, pH=5.8 cultured for 20 days to induce embryonic leaflets to produce clustered buds or clustered bud tissue blocks to obtain tissue cultured plantlets;

[0029] (3) Cut out the tissue culture seedlings obtained i...

Embodiment 2

[0036] A method for culturing immature embryos of red-skinned peanuts, the technical scheme of which comprises the following steps:

[0037] (1) Peel the mature pods of peanuts, select the seeds with full particles and smooth surface, rinse with water, absorb the surface water, and then soak them in 75% ethanol solution by volume for 3 minutes, and mercuric chloride with 0.1% by mass solution for 13 minutes, then rinsed with sterile water for 8 times, then peeled off the seed coat of peanut seeds, soaked in sterile water for 10 hours, to obtain sterile peanut embryo leaflets;

[0038] (2) Take the sterile peanut embryo leaflets obtained in step (1) and inoculate them on the cluster bud induction medium, the cluster bud induction medium is MS+0.3mg / L NAA+7mg / L6-BA, pH=5.8 cultured for 25 days to induce embryonic leaflets to produce clustered buds or clustered bud tissue blocks to obtain tissue cultured plantlets;

[0039] (3) Cut out the tissue culture seedlings obtained in st...

Embodiment 3

[0046] A method for culturing immature embryos of red-skinned peanuts, the technical scheme of which comprises the following steps:

[0047] (1) Peel the mature pods of peanuts, select the seeds with full particles and smooth surface, rinse with water, absorb the surface water, and then soak them in 75% ethanol solution by volume for 2 minutes, and 0.1% by mass of mercuric chloride soak in the solution for 11 minutes, then rinse with sterile water for 6 times, peel off the seed coat of the peanut seeds, soak in sterile water for 8 hours, and obtain sterile peanut embryo leaflets;

[0048] (2) Take the sterile peanut embryo leaflets obtained in step (1) and inoculate them on the cluster bud induction medium, the cluster bud induction medium is MS+0.2mg / L NAA+6mg / L6-BA, pH=5.8 cultured for 23 days to induce embryonic leaflets to produce clustered buds or clustered bud tissue blocks to obtain tissue cultured plantlets;

[0049] (3) Cut out the tissue-cultured seedlings obtained ...

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Abstract

The invention belongs to the field of plant-tissue culture methods, and particularly relates to an immature-embryo culture method for red-peel peanuts. The immature-embryo culture method based on the existing peanut tissue culture comprises the following steps: disinfection and soaking of seeds, induced culture of clustered shoots, culture of strong tissue culture seedlings, rooting culture of seedlings, and transplantation after seedling hardening of complete plants. The immature-embryo culture method has the advantages that complete peanut plants can be obtained within 15 weeks by carrying out tissue culture on peanuts, so the period for tissue culture is greatly shortened, mass propagation in short time can be achieved, and the significance for propagation of rare varieties and the current peanut transgenosis is very important; secondly, culture conditions can be artificially controlled, and are not limited by regions and seasons, so that convenience is brought for stable culture production, and foundation is laid for development of tissue culture and genetic transformation of peanuts.

Description

technical field [0001] The invention belongs to the field of plant tissue culture methods, in particular to a method for culturing immature embryos of red-skinned peanuts. Background technique [0002] Peanut is an important economic crop and oil crop in the world. my country's peanut production ranks first in the world, and its planting area ranks second in the world. It plays a pivotal role in the world's peanut production. Traditional breeding can effectively improve the characteristics of peanuts in terms of yield and drought resistance, but genetic engineering breeding can more effectively retain and screen some beneficial purpose traits than traditional breeding methods. The genetic basis of peanut is weak, and the available effective resources are limited. The key to genetic transformation of peanut using genetic engineering technology is to establish an efficient receptor system, that is, to establish an efficient method for tissue culture of peanut plants. [0003...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/005A01H4/001A01H4/008
Inventor 卢海凤
Owner GUANGXI UNIV
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