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Method for culturing wheat tissue capable of overcoming gene distrubance

A tissue culture and genotyping technology, applied in horticultural methods, botanical equipment and methods, plant phenotype improvement, etc. Genotypic disorders, the effect of increasing the frequency of regeneration, and improving the efficiency of tissue culture

Inactive Publication Date: 2004-06-09
INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Most previous researchers only selected one or several breed genotypes as materials. Due to the small number of breeds studied and not representative, these studies are difficult to answer and solve the problem of genotype barriers

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Response of young ears at different developmental stages to in vitro culture

[0022] A plant material: 5 varieties (lines) of 6756, 8581, H89, 311 and 1772 were selected.

[0023] B sampling period: the two-edge stage (1mm), the glume primordium formation stage (2mm), the floret primordium formation stage (2.5mm), the pistil primordium formation stage (4mm), and the connective differentiation stage (6mm) and the young ears of the tetrad formation stage (10mm) stage are carried out in vitro culture, wherein, the young ears of the drug interval stage and the tetrad stage are cut into 2-3 sections for culture. 0.1% HgCl before inoculation 2 Disinfect for 8-10 minutes, rinse with sterile water 3-5 times, use tweezers and a dissecting needle to peel off the young ears, and inoculate them on the induction medium.

[0024] C medium

[0025] The induction medium is MS+2, 4-D (2mg / l);

[0026] The differentiation medium is MS basic medium;

[0027] Sprout elongat...

Embodiment 2

[0059] The response of different explants of embodiment 2 to in vitro culture

[0060] The material selection and cultivation method of A suitable young ear are the same as above;

[0061] B Immature Embryo Selection and Culture Method

[0062] Operation steps: a. Take immature embryos 14 days after flowering, and use 0.1% HgCl 2 Disinfect for 8-10 minutes, rinse with sterile water 3-5 times, use tweezers and a dissecting needle to peel off immature embryos, inoculate on induction medium and culture in dark for 25-30 days; b. Transfer callus to differentiation medium Differentiate (25-30 days); c. Green seedlings are transferred to strong root medium.

[0063] Medium and culture conditions: induction medium is MS+2, 4-D (2mg / 1); differentiation medium is MS basic medium; strong root medium is 1 / 2MS+IAA2mg / l. In the callus induction period, culture in the dark at room temperature (25±1)°C; in the green seedling differentiation stage, culture in the room temperature (26±1)°C ...

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Abstract

The present invention relates to a wheat tissue culture method overcoming genotypic disturbance. Said invention selects the young-ear as proper explant, and finds out optimum development stage of young-gear which is glume-protecting differentiation period to pistil and stamen primordial differnetiation period, and researches the all proper culture media and culture conditions of different culture periods. Said invention utilizes the selection of proper explant and regulation of culture medium component, and matches the improved measure so as to can effectively overcome the genotypic disturbance problem in wheat tissue culture, and can greatly raise the plant regeneration frequency and tissue culture efficiency.

Description

Technical field: [0001] The invention relates to a wheat tissue culture method for overcoming genotype barriers. Background technique: [0002] At present, the research on transgenic wheat is basically in the stage of establishing and optimizing the transgenic system, which is far behind that of rice and corn. One of the main reasons is the low plant regeneration frequency and genotype barriers of wheat transgenic recipients, which have become the bottleneck limiting the success of wheat transgenics. For wheat varieties with high plant regeneration frequency, transgenic plants have been obtained no matter by biolistic method or Agrobacterium method, while for varieties with low plant regeneration frequency, transgenic plants cannot be obtained even by biolistic method. It is precisely because of the problem of regeneration frequency caused by the genotype difference of varieties that most of the research on transgenic wheat in the world is concentrated on a few genotypes su...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H3/02A01H4/00C12N5/04
Inventor 刘录祥赵林姝郑企成王晶赵世荣郭会君陈文华
Owner INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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