Ploidy appraising method of isolated microspore regeneration plant strain of non-heading Chinese cabbage

A technology for regenerating plants and head cabbage, which is applied in the directions of instruments, fluorescence/phosphorescence, and analytical materials, etc., to achieve the effects of simple sample preparation, high resolution and accuracy, and fast testing speed.

Active Publication Date: 2008-03-05
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

It has been successfully used to identify the ploidy of cauliflower (CHIANG, 1985; ROULUND, 1990), broccoli (WANG, 1999), Brussels sprouts (Zhou Yuanchang, 2002), and Chinese

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  • Ploidy appraising method of isolated microspore regeneration plant strain of non-heading Chinese cabbage

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Embodiment Construction

[0024] A method for identifying the ploidy of non-heading cabbage free microspore regeneration plants provided by the present invention, comprising:

[0025] 1. Materials and methods: Free microspore culture was carried out on the excellent non-heading Chinese cabbage hybrid 'Sulu' (Geng Jianfeng, 2007), and 156 microspore regenerated plants were obtained. The material used was 'Sulu', which was sold in the market; in Nanjing The Cytogenetics Workstation of the College of Horticulture of the Agricultural University adopts the Epics XL flow cytometer produced by Beckman Coulter Company of the United States to conduct flow cytometry ploidy analysis on the test tube seedlings. The applied analysis software is EXPO32ADCsoftware.

[0026] After the microspore regenerated plants bolted and flowered, the adult morphological identification of all regenerated seedlings was carried out. Haploid plants have slender branches, many branches, small flower buds, petals and anthers, no polle...

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Abstract

The method comprises: preparing single cell suspension liquid; using a flow cytometer produced by US BeckMAN COULTER company to make the flow cytometer ploidy analysis; using 488nm argon ion air cooling laser to measure the DNA molecule fluorescence intensity; if the peak value of the plant to be measured is close to 140, it is considered as a diploid; if it is close to 70 and 280, it is considered as haploid and tetraploid.

Description

1. Technical field [0001] The invention relates to a method for identifying ploidy of non-heading cabbage free microspore regenerated plants, which belongs to the field of biotechnology and is specially used for rapid identification of non-heading cabbage free microspore regenerated plant ploidy. 2. Technical background [0002] Non-heading cabbage (Brassica campestris ssp.chinensis Makino), native to China, is the most popular vegetable widely grown in southern my country, and plays a pivotal role in the annual supply of vegetables. Non-heading Chinese cabbage is a typical cross-pollination crop, with severe selfing decline and obvious advantages of excellent hybrid combinations. The microspore culture technology using the haploid approach can accelerate the purification of breeding materials, shorten the breeding cycle, and accelerate the breeding process. The regenerated plant populations obtained through free microspore culture include, in addition to the double single ...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N15/00
Inventor 侯喜林成妍
Owner NANJING AGRICULTURAL UNIVERSITY
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