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Method for quickly culturing barley anther and used culture media

The technology of a culture medium and induction medium, which is applied in the field of plant tissue culture, can solve the problems of increasing the cost and workload of barley anther culture, limiting the application of barley anther culture technology, and the regeneration rate of green shoots being less than 3%, so as to shorten the time of tissue culture. cycle, usage reduction, and the effect of reducing workload

Inactive Publication Date: 2012-07-18
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since barley anther culture is one of the most difficult crops to induce plant regeneration, the green shoot regeneration rate of most genotypes is still less than 3% so far.
And the currently used barley anther culture method generally passes through 4 steps of cultivation, followed by anther pretreatment, induced callus culture, differentiation culture and rooting and strong seedling culture, and the latter 3 steps all need to prepare corresponding medium, which increases the barley anther The cost and workload of cultivation, and the cultivation time used is longer, generally it takes about 15 weeks to complete the whole cultivation process
These all limit the application of barley anther culture technology

Method used

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  • Method for quickly culturing barley anther and used culture media
  • Method for quickly culturing barley anther and used culture media

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1, a kind of barley anther medium, is made up of pretreatment medium, induction medium, differentiation / rooting medium:

[0040] 1), pretreatment medium:

[0041] Mannitol: 182g / L, Agar: 15g / L, the rest is distilled water.

[0042] The preparation method is as follows: Weigh 182 g of Mannitol and 15 g of agar (Agar), dilute to 1 L with distilled water; then sterilize at 1.1 atmospheres and 121° C. for 20 minutes. Pour it into a 75x15mm sterilized Petri dish, seal it with a parafilm after solidification, and store it at room temperature for later use.

[0043] 2), induction medium:

[0044] Its preparation method is as follows:

[0045] I), glutamine, inositol, VB 1 , IAA and BAP were prepared with distilled water respectively, and after 0.2 μm microporous membrane filtration sterilization (1.1 atmospheric pressure, sterilized at 121° C. for 20 min), glutamine solution (concentration was 750 mg / ml) and inositol solution ( Concentration is 100mg / ml), VB 1...

Embodiment 2

[0053] Example 2, a set of methods for culturing barley anthers, using the corresponding medium in Example 1, using the barley hybrid combination "Gairdner / Zhebei 33" F 1 Generation (this material is planted in the field under natural conditions) is the material and carries out the following steps successively:

[0054] 1) Take the anthers before the barley ears have bloomed, and observe the growth period of the anthers under a microscope by staining with acetic acid magenta. Take the ears of wheat in this period in the field, disinfect them with 70% (v / v) alcohol on the aseptic operation table, and after drying, take out the ears in the leaf sheath with a sterilized blade, and then use a pointed Head tweezers take out the anthers in the ears, put them on the pretreatment medium, put 60-70 anthers in each petri dish, and seal the parafilm. Cultivate for 3-5 days in a dark environment at 20-25°C.

[0055] 2), transfer the cultured anthers obtained in step 1) to the induction ...

Embodiment 3

[0060] Embodiment 3, according to the method of embodiment 2, to the F of hybrid combination "Kunlun 12 / flower 30" 1 A total of 500 anthers were cultivated, and 77 green shoots were obtained, and the regeneration rate of green shoots reached 15.4%.

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Abstract

The invention discloses a barley anther culture medium, which comprises a pretreatment culture medium, an inducing culture medium and a differentiating / rooting culture medium. The invention further provides a method for rapidly culturing barley anther by using the barley anther culture medium. The method comprises the following steps of: (1) taking a wheat ear of which a microspore develops at a mononuclear metaphase from a field, taking anther out of the wheat ear, putting onto the pretreatment culturing medium, and culturing in a dark environment for 3-5 days; (2) transferring the pretreated anther onto the inducing culture medium, and culturing in a dark environment for 4-5 weeks; (2) transferring callus and a seedling obtained by inducing in the step (2) onto the differentiating / rooting culture medium, performing illumination culturing at the temperature of 22-25 DEG C and at the illumination intensity of 40-60 mumol / m<2> / s for 3-4 weeks, 12 hours every day, to obtain a rooting green seeding. Due to the adoption of the method disclosed by the invention, the culturing efficiency of the barley anther can be increased remarkably.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to an optimized culture medium for barley anther culture and a simple and rapid culture method thereof. Background technique [0002] Barley is the fourth largest cereal crop in the world. Its planting area and total output are second only to wheat, rice and corn. It is mainly used in animal feed, malt manufacturing, food manufacturing and medicine. Breeding fine varieties is an important part of barley breeding research. Target. Barley anther culture is a new technique for haploid breeding, which can simplify the breeding procedure and speed up the breeding process. The time required for doubling haploid breeding to produce pure lines usually saves 3-4 generations compared with the time required for conventional breeding , and the selection of qualitative traits and quantitative traits is more reliable and effective, and the doubled haploid population is a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 杨建明华为汪军妹贾巧君朱靖环尚毅林峰顾玉坤
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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