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Tissue culture method of young barley embryos directly into seedlings and the medium used

A tissue culture medium and culture medium technology, applied in the field of plant tissue culture, can solve the problems of high contamination rate, easy variation, long callus-differentiation-seedling process cycle, etc.

Active Publication Date: 2016-04-13
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method can solve the current problems of time-consuming barley seed maturation and breaking dormancy, traditional barley immature embryo tissue culture: long callus-differentiation-seedling process cycle, high pollution rate, easy to mutate, etc.

Method used

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  • Tissue culture method of young barley embryos directly into seedlings and the medium used

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1, barley immature embryo culture medium in vitro, is made up of these 2 types of culture medium of seedling medium and rooting strong seedling medium:

[0050] 1), seedling growth medium:

[0051] First prepare the mother liquor of the seedling medium:

[0052] The mother liquor of seedling growth medium is composed of the following components:

[0053] KNO 3 1900mg / L, NH 4 NO 3 165mg / L, KH 2 PO 4 170mg / L, CaCl 2 2H 2 O165mg / L, MgSO 4 ·7H 2 O185mg / L, Na 2 -EDTA·2H 2 O37.3mg / L, FeSO 4 ·7H 2 O27.8mg / L, H 3 BO 3 6.2mg / L, MnSO 4 4H 2 O22.3mg / L, ZnSO 4 ·7H 2 O8.6mg / L, Na 2 MoO 4 2H 2 O0.25mg / L, KI0.83mg / L, CuSO 4 ·5H 2 O0.025mg / L, CoCl 2 ·6H 2 O0.025mg / L, proline 600mg / L, glutamine 250mg / L, hydrolyzed casein 400mg / L, inositol 100mg / L and maltose 30g / L, the rest is distilled water;

[0054] The preparation method of the mother liquor of seedling growth medium is as follows:

[0055] ①. Weigh KNO separately 3 19.0g, NH 4 NO 3 1.65g, K...

Embodiment 2

[0068] Embodiment 2, a method for in vitro culture of barley immature embryos, using the corresponding medium in Example 1, with "Zhexiu 12 / Subei No. 3" F 1 Barley was used as the experimental material in the following steps:

[0069] 1) Select "Zhexiu 12 / Su Beer No. 3" F 1 The immature seeds of barley 12-15 days after pollination, peel off the shell to expose the embryo, then soak the above-mentioned treated seeds in 50% (mass concentration) sodium hypochlorite solution for 30 minutes, discard the sodium hypochlorite solution and add 70% (v / v) Soak in alcohol for 5 minutes, then discard 70% alcohol and add 3% (v / v) hydrogen peroxide to soak for 5 minutes, and finally wash 5 times with sterile water.

[0070] On the aseptic operating table, use a sterilized scalpel to gently puncture the sterilized immature seeds from the side of the embryo, then slowly take out the embryo from the front end of the embryo and inoculate it into the on seedling medium.

[0071] 2) Place the ...

Embodiment 3

[0077] Example 3. According to the method of Example 2, the immature embryos of the barley variety "Zhepi 33" were cultured in vitro. Three repetitions were set up, and 60 embryos were inoculated each time. The seedling rate (average value) was 90.6%.

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Abstract

The invention discloses a tissue culture medium for directly growing a seedling from a barley embryo. The tissue culture medium comprises a seedling growing culture medium and a root growing and seedling strengthening culture medium. A preparation method of the seedling growing culture medium comprises the following steps: firstly, regulating the pH value of every 1L of mother solution of the seedling growing culture medium to be 5.7-5.9; secondly, adding plant gel; thirdly, performing sterilization; and finally, adding filtered and sterilized vitamin B1, vitamin B6, nicotinic acid, 6-benzylaminopurine and 2,4-dichlorphenoxyacetic acid. A preparation method of the root growing and seedling strengthening culture medium comprises the following steps: firstly, adding inositol, casein hydrolysate, proline and malt sugar into every 1L of MS basic culture solution; secondly, regulating the pH value to be 5.8-6.0, then adding the plant gel and then performing the sterilization; and finally, adding the filtered and sterilized vitamin B1. The invention also provides a tissue culture method for directly growing the seedling from the barley embryo, and in the method, tissue culture medium is used.

Description

technical field [0001] The invention belongs to the field of plant tissue culture, and in particular relates to a tissue culture method and a corresponding tissue culture system of barley immature embryos that rapidly grow into seedlings. Background technique [0002] Barley is one of the main food crops in the world and can be used as beer, feed, food and health products. It takes too long to cultivate new barley varieties by conventional breeding, and it takes about 8 to 10 years without additional generations. In order to shorten the breeding period, breeders have explored and studied many methods of adding generations. In summary, there are mainly the following methods: one is to choose to use the natural environment and season of the barley ecological type that is geographically more suitable for breeding in different places; the other is to use natural In situ or off-site adding generations combined with natural conditions and artificial climate chambers; the third is...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04A01H4/00
Inventor 尚毅杨建明华为朱靖环汪军妹贾巧君
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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