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Short-period tissue culture method of peanuts

A tissue culture, short-cycle technology, applied in horticultural methods, botanical equipment and methods, plant regeneration, etc., can solve the problems of difficulty in growing plants, large demand for seeds, low repeatability, etc., and shorten the cycle required for tissue culture. , speed up the breeding speed, good repeatability

Inactive Publication Date: 2013-06-26
HEBEI AGRICULTURAL 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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  • Short-period tissue culture method of peanuts
  • Short-period tissue culture method of peanuts
  • Short-period tissue culture method of peanuts

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The screening of embodiment 1 cluster bud induction medium

[0040] The present embodiment takes kind sesame oil 1-1 as material.

[0041] The basic medium used in this example is MSB 5 Medium, including the large amount, trace amount and iron salt components in MS medium and B 5 For the culture medium of organic components in the culture medium, its mother liquor configuration is shown in Table 1-Table 5:

[0042] Table 1MS a large number of mother liquor (20 times) configuration

[0043]

[0044] Table 2 MS trace mother solution (200 times) configuration

[0045]

[0046] Table 3 MS iron salt mother liquor (200 times) configuration

[0047]

[0048] Table 4B 5 Organic element mother solution (1000 times) configuration

[0049]

[0050]

[0051] Table 5 Inositol mother liquor (100 times) configuration

[0052]

[0053] MSB per liter 5 The medium contained 30 g of sucrose, 7 g of agar, and had a pH of 5.8.

[0054] to the above MSB 5 The medium...

Embodiment 2

[0065] The screening of embodiment 2 elongation medium

[0066] This embodiment uses the MSB in Embodiment 1 5 The medium is the basic medium, which is formulated according to the combination in Table 8 and added with 6-benzylpurine (6-BA), α-naphthaleneacetic acid (NAA), gibberellin (GA 3 ), to obtain the elongation medium to be screened, with a pH value of 5.8.

[0067] Table 8 The hormone type and concentration combination of the elongation medium to be selected

[0068]

[0069] In this example, the clustered buds or clustered bud tissue pieces induced in Example 1 were cut, transferred to 7 elongation mediums to be tested, and placed in a light incubator for cultivation, with light intensity of 1500-2000lx, time of 16h / d, and temperature 26±0.5°C, subculture once every three weeks, the subcultured seedlings are as follows: image 3 As shown, there are both seedlings and newly differentiated cluster buds on the side, which are used to elongate and proliferate the clu...

Embodiment 3

[0075] Embodiment 3 Establishment and verification of peanut short-cycle tissue culture method of the present invention

[0076] According to the result of embodiment 1-2, establish peanut short cycle tissue culture method of the present invention, comprise the steps:

[0077] 1) Surface disinfection and soaking of peanut seeds;

[0078] 2) Taking the embryonic leaflets of the peanut seeds described in step 1), inoculating them on the cluster bud induction medium for culture, and inducing the embryonic leaflets to produce cluster buds or cluster bud tissue blocks;

[0079] The cluster bud induction medium is MSB 5 +0.2mg / L NAA+5~6mg / L6-BA, solid medium with pH=5.8;

[0080] 3) Cut out the clustered buds or clustered bud tissue pieces described in step 2), inoculate them on the No. 1 elongation medium for culture, and subculture once every three weeks; cut newly differentiated clustered buds or clustered bud tissue pieces for each subculture and go to step 2) Cultivate on th...

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Abstract

The invention provides a short-period tissue culture method of peanuts. The short-period tissue culture method comprises the following steps of: (1) disinfecting the surfaces of peanut seeds and soaking the seeds; (2) inoculating embryonic leaflets of the peanut seeds on a cluster-bud inducing medium; (3) cutting cluster buds or cluster-bud tissue blocks, inoculating the cluster buds or the cluster-bud tissue blocks on a first extending medium for culture and a second extending medium for alternative culture till seedlings of 2cm-3cm in length are obtained; (4) cutting the seedlings of 2cm-3cm in length and inoculating the cut seedlings on a rooting medium for culture till strong roots grow and complete plants are obtained; and (5) hardening the complete plants and transplanting the hardened complete plants onto vermiculite. The short-period tissue culture method provided by the invention is used for carrying out tissue culture on peanuts, rooted peanut tissue culture seedlings can be obtained in 15 weeks, and the period of tissue culture is greatly shortened, so that the breeding speed of the peanut tissue culture seedlings is enhanced; and the peanut tissue culture seedlings cultured by the method are high in regeneration rate and good in repeatability.

Description

technical field [0001] The invention belongs to the field of plant tissue culture, and in particular relates to a peanut short-cycle tissue culture method. 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] Peanut is a ki...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 刘立峰马彩霞穆国俊侯名语陈焕英何美敬
Owner HEBEI AGRICULTURAL UNIV.
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