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A method for repeatedly regenerating leaves of test-tube plantlets of wheat

A technology for test-tube seedlings and wheat, applied in the field of plant tissue culture, can solve the problems of repeated regeneration of wheat test-tube seedlings and other problems

Inactive Publication Date: 2011-11-30
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the deficiencies in the prior art, the purpose of the present invention is to provide a method for repeated regeneration of young leaves of wheat test-tube plantlets to solve the problem of repeated regeneration of wheat test-tube plantlets.

Method used

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  • A method for repeatedly regenerating leaves of test-tube plantlets of wheat
  • A method for repeatedly regenerating leaves of test-tube plantlets of wheat
  • A method for repeatedly regenerating leaves of test-tube plantlets of wheat

Examples

Experimental program
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Effect test

Embodiment 1

[0048] The immature ears of Jinan 177 wheat 14 days after flowering were taken and sterilized by immersing them in 75% alcohol by volume for 5 minutes. Peel the immature wheat grains from the ears and place them in a sterile petri dish, clamp them with the ventral groove downward, and pick out the immature embryos with a dissecting needle. Select immature embryos with a diameter of 1mm, and inoculate them with the scutellum up in MS basic components + B5 vitamins + 146mg / L glutamine + 200mg / L hydrolyzed casein + 1mg / L 2,4-D + 7g / L agar + 30g / L Sucrose callus induction medium I (pH 5.8), 25 ℃, dark culture. The embryogenic callus was obtained in 2 weeks, and the induction rate reached 100%.

[0049] Excise the embryo directly grown from the immature embryo on the induced callus, and then transfer the induced callus to MS basic component + B5 vitamin + 146mg / L glutamine + 2mg / L 2,4-D+ On the embryogenic callus subculture medium of 7g / L agar+30g / L sucrose (pH value 5.8), the li...

Embodiment 2

[0056] The immature ears of Chinese spring wheat 15 days after flowering were taken and sterilized by soaking them in 75% alcohol for 3 minutes. Peel the immature wheat grains from the ears and place them in a sterile petri dish, clamp them with the ventral groove downward, and pick out the immature embryos with a dissecting needle. Select immature embryos with a diameter of 0.8mm, and inoculate them with MS basic components + B5 vitamins + 130mg / L glutamine + 180mg / L hydrolyzed casein + 0.5mg / L 2,4-D + 6g / L agar + 25g with the scutellum up / L sucrose callus induction medium I (pH 5.8), 25 ℃, dark culture. The embryogenic callus was obtained in 2 weeks, and the induction rate reached 99%.

[0057] Excise the embryo directly grown from the immature embryo on the induced callus, and then transfer the induced callus to MS basic ingredient + B5 vitamin + 130mg / L glutamine + 1mg / L 2,4-D+ On the embryogenic callus subculture medium of 6g / L agar+25g / L sucrose (pH 5.8), the light in...

Embodiment 3

[0064] The immature ears of Wheat No. 3 were taken 16 days after flowering and soaked in 75% alcohol for 8 minutes to sterilize. Peel the immature wheat grains from the ears and place them in a sterile petri dish, clamp them with the ventral groove downward, and pick out the immature embryos with a dissecting needle. Select immature embryos with a diameter of 1.2mm, and inoculate them with MS basic components + B5 vitamins + 170mg / L glutamine + 220mg / L hydrolyzed casein + 2mg / L 2,4-D + 8g / L agar + 35g / L sucrose on callus induction medium I (pH 5.8), cultured at 25°C in the dark. The embryogenic callus was obtained in 2 weeks, and the induction rate reached 100%.

[0065] Excise the embryo directly grown from the immature embryo on the induced callus, and then transfer the induced callus to MS basic component + B5 vitamin + 170mg / L glutamine + 2mg / L 2,4-D+ 8g / L agar + 35g / L sucrose on the subculture medium of embryogenic callus (pH 5.8), carry out day and night light cycle c...

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Abstract

The invention relates to a method for repeatedly regenerating wheat test-tube plantlets through tissue culture of wheat test-tube plantlet leaves, belonging to the technical field of plant tissue culture, and the steps include: (1) induction of embryogenic callus; (2) callus (3) Acquisition of test-tube plantlets; (4) Induction of callus from leaves of test-tube plantlets; (5) Induction of secondary regenerated plantlets; (6) Induction of rooting; (7) Transplantation of secondary regenerated plantlets . The method for repeated regeneration of young leaves of wheat test-tube plantlets provided by the present invention successfully solves the problem of repeated regeneration of wheat test-tube plantlets. It is possible to obtain homogeneous chloroplast transgenic wheat. According to test statistics: the frequency of callus induced by leaves of test-tube plantlets using the method and the frequency of secondary regenerated seedlings have reached more than 93% and 66% respectively.

Description

technical field [0001] The invention relates to a method for regenerating leaves of test-tube plantlets of wheat, in particular to a method for repeatedly regenerating test-tube plantlets of wheat through tissue culture of leaves of test-tube plantlets of wheat, and belongs to the technical field of plant tissue culture. Background technique [0002] Wheat (Triticum aestivum L) is an important economic crop in my country and one of the main sources of food. In order to improve wheat, the current wheat transgenic research is to transfer the target gene into the nucleus. Compared with nuclear transgenes, chloroplast transgenes have the advantages of high expression, genetic stability, and environmental friendliness. As a new direction of crop genetic improvement, chloroplast genetic engineering has not been applied to the improvement of wheat genetic traits. This is because it is difficult for cereal crops such as wheat to regenerate repeatedly with green leaf tissue, which ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04A01H4/00
Inventor 侯丙凯王文超耿晓霞胡洪群
Owner SHANDONG UNIV
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