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Method for increasing barley microspore culture callus yield and green seedling yield

A microspore culture and barley technology, applied in the field of bioengineering, can solve problems such as the limitation of the area of ​​an artificial climate chamber, the limitation of the number of planted barley seedlings, etc., and achieve the effects of increasing the number of green seedlings, increasing the yield of callus, and optimizing the cultivation process.

Inactive Publication Date: 2018-08-17
SHANGHAI ACAD OF AGRI SCI
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  • Description
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Problems solved by technology

However, due to economic reasons, the area of ​​the artificial climate chamber is limited, and the number of barley seedlings to be planted is greatly limited.

Method used

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  • Method for increasing barley microspore culture callus yield and green seedling yield
  • Method for increasing barley microspore culture callus yield and green seedling yield

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Embodiment 1 low temperature treatment barley material flower 30 seedlings

[0020] Plant barley material flower 30 in the pot (barley variety flower 30 is an excellent barley variety cultivated by the cell engineering laboratory of Shanghai Academy of Agricultural Sciences Institute of Biotechnology, the domestic planting area is relatively wide, the seeds are provided by the cell engineering laboratory of Shanghai Academy of Agricultural Sciences Preservation), when the seedlings grow to the stage of two leaves and one heart, they are moved to the intelligent incubator for low temperature treatment. The plant cultivation conditions are: temperature 4°C at night and 4°C during the day; humidity 65%; 12 hours of light and 12 hours of darkness. After 30 days of low temperature treatment, they were transplanted into the artificial climate chamber, and the plants without low temperature treatment were planted as the control group at the same time. The plant culture conditio...

Embodiment 2

[0021] Embodiment 2 barley material flower 30 microspore culture

[0022] Select low-temperature treatment and control barley ears whose florets and microspores in the middle of the young ears of the control plants are in the early-middle stage of uninucleation, cut off the leaves (keep the base of the upper two leaves about 1.5 cm), wrap the young ears with clean wet gauze, and put them in Moisturize in a fresh-keeping bag, mark the date and quantity of the material, and put it in a 4°C refrigerator for 2 to 4 weeks of low-temperature pretreatment. Free microspores from flower buds were taken from low-temperature-treated materials, and the microspores were freed with reference to methods such as Lu Ruiju (references: Lu Ruiju, Huang Jianhua, etc., the impact of colchicine on the survival and seedling formation of barley in vitro cultured microspores. Journal of Plant Physiology, 2001 , 27(2): 135-140). Each test tube connects the flower buds of 4 spikes, and adds 12ml extrac...

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Abstract

The invention relates to the technical field of bioengineering, in particular to a method for increasing barley microspore culture callus yield and green seedling yield. The method includes steps: planting barley, and subjecting to low-temperature treatment when seedlings grow to a two-leaf period, namely transplanting in a phytotron after culture at 3-5 DEG C for 28-32 days. The method has advantages that improvement is performed aiming at seedling management in a barley microspore technique, barley seedlings are subjected to low-temperature treatment for a period of time and then transplanted in the phytotron, and accordingly barley microspore culture callus yield and green seedling yield are remarkably increased, and high efficiency in operation of the barley microspore technique is realized. The improvement technique can be applied to barley haploid breeding and haploid transformation.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for improving the callus yield and green shoots of barley microspore culture. Background technique [0002] Plant microspores (Microspores) are haploid cells produced by tetrads formed by pollen mother cells after meiosis. Embryos develop into complete plants. Using this haploid cell culture system for gene transformation, homozygosity can be quickly stabilized through chromosome doubling, and high-frequency regeneration plants can be obtained at the same time. [0003] Microspore in vitro culture technology is an important haploid breeding technology developed at present, and a mature free microspore culture system has been established in various crops such as tobacco, rape, corn, and pepper. Using field planting materials as donor plants, our laboratory has established a domestically leading and internationally advanced high-efficiency microspore regeneration ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 李颖波何婷郭桂梅刘成洪陆瑞菊黄剑华
Owner SHANGHAI ACAD OF AGRI SCI
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