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Method for rapid identification of Gerbera jamesonii ploidy by using flow cytometry

A flow cytometry, rapid technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problem of no ploidy identification report, achieve fast test speed, high sensitivity and resolution, and make the simple effect

Inactive Publication Date: 2012-06-27
FLOWER RES INST OF YUNNAN ACAD OF AGRI SCI
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  • Application Information

AI Technical Summary

Problems solved by technology

At present, it has been successfully used in the ploidy identification of cauliflower, broccoli, Brussels sprouts, grapes, Chinese cabbage, and kiwifruit plants. It has special requirements for ploidy identification, so there is no report on ploidy identification of gerbera megaspore regenerated plants by flow cytometry

Method used

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  • Method for rapid identification of Gerbera jamesonii ploidy by using flow cytometry
  • Method for rapid identification of Gerbera jamesonii ploidy by using flow cytometry
  • Method for rapid identification of Gerbera jamesonii ploidy by using flow cytometry

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Experimental program
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Effect test

Embodiment 1

[0038] Example 1 Ploidy identification of gerbera megaspore regenerated plants

[0039] 1. Preparation of materials and sample processing preparations

[0040] ① Sample material to be tested: gerbera variety 'Ju Pai Wang', which is derived from the gerbera germplasm resource preservation nursery of the Institute of Flowers, Yunnan Academy of Agricultural Sciences, and was induced in vitro by Wang Lihua et al. and plant regeneration [J]. Physiological Communications, 2007, 43 (6) 1089-1092) to induce megaspore culture, and obtain 24 megaspore regeneration plants. Cytometry, flow cytometry ploidy analysis of megaspore regenerated plants. The applied analysis software is Partec CyView 85 software.

[0041] ② Control sample material: conventional test-tube seedlings of gerbera variety 'Ju Pai Wang', identified as diploid (2n=2x=50) material by root tip chromosome.

[0042] ③Preparation of sample treatment preparation: The sample treatment preparation consists of extraction and ...

Embodiment 2

[0055] Example 2 Ploidy identification of test-tube plantlets obtained from regenerated haploid plants of Gerbera chrysanthemum megaspores treated with colchicine.

[0056] 1. Materials and methods

[0057] ① Sample material to be tested: gerbera variety 'Savannah', which comes from the gerbera germplasm resource preservation nursery of the Yunnan Academy of Agricultural Sciences Flower Institute, using Wang Lihua et al. Regeneration [J]. Physiological Communications, 2007, 43(6) 1089-1092) to induce megaspore culture and obtain 11 regenerated plants with megaspores. After root tip chromosome identification, 3 haploid plants were obtained. Propagate these three haploid strains to more than 50 strains respectively, and place them in the propagation medium MS+BA 0.6 mg.L supplemented with 0.025% colchicine -1 +NAA 0.1 mg.L -1 (pH 5.8) for 6 days. Transfer to MS+BA 0.3 mg.L after treatment -1 +NAA 0.1 mg.L -1 (pH 5.8) culture medium for 30-50 days. The 114 normal survivin...

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Abstract

The invention discloses a method for rapid identification of Gerbera jamesonii ploidy by using flow cytometry. The method comprises steps of: preparing a unicell fluid suspension by using an extraction cracking buffer and a dyeing buffer; determining DNA fluorescence intensity of a sample by using a CyFlow Cube type of flow cytometer produced by German partec company and carrying out ploidy analysis; and comparing the abscissa positions of the sample to be measured and the contrast DNA content fluorescence intensity separation peak to identify the ploidy of the sample to be measured. A flow cytometer ploidy identification result is consistent with root tip chromosome counting ploidy. According to the method, ploidy of batch of Gerbera jamesonii macrospore regeneration plant can be screened out rapidly, and efficiency of DH colony construction and heredity atlas drafting is increased substantially, so as to accelerate a breeding process. The invention has advantages of simple sample preparation, rapid detection and stable high detection rate.

Description

technical field [0001] The invention relates to a method for quickly identifying the ploidy of gerbera by flow cytometry, which belongs to the field of biotechnology and is specially used for the rapid identification of ploidy of gerbera megaspore regenerated plants. Background technique [0002] Gerbera ( Gerbera jamesonil Bolus), also known as Gerbera, is a plant of the genus Gerbera in the family Asteraceae. It has bright colors and can be used for cut flower production or potted ornamental. It is widely distributed all over the world and is a flower with high ornamental value. At present, there is an urgent need for high-quality and high-yield new varieties of gerbera that are resistant to low temperature, low light, blight and viral diseases. However, the conventional breeding cycle is long and the range of variation spectrum is limited, so it is difficult to capture the target traits. Obtaining double haploids through pollen (anther) culture is a method to efficientl...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 王丽花瞿素萍杨秀梅吴学尉张丽芳张艺萍苏艳彭绿春
Owner FLOWER RES INST OF YUNNAN ACAD OF AGRI SCI
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