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Slide preparation method for discriminating chromosome number of Avena magna, Triticum aestivum or filial generation of Avena magna and Triticum aestivum

A chromosome number and tetraploid technology, applied in the biological field, can solve the problems of hypotonicity after lack, chromosome sticking together, poor chromosome dispersion, etc., and achieve the effect of good chromosome dispersion, more cell division phases, and high repeatability

Inactive Publication Date: 2016-04-20
CAPITAL NORMAL UNIVERSITY
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AI Technical Summary

Problems solved by technology

[0019] First, after pretreatment with saturated α-bromide, which is commonly used by researchers, at room temperature for 3 hours or with distilled water at 4°C for overnight treatment, there are fewer mitotic phases in the root tip cells, and the chromosomes stick together, making it difficult to identify
Direct hypotonicity and fixation without any pretreatment, there are more mitotic phases in the root tip cells, but the chromosome dispersion is poor
[0020] Second, hypotonic treatment with distilled water or 0.075MKCl at room temperature for 30 minutes cannot achieve the effect of dispersing chromosomes
[0021] Third, it is difficult to soften the root tip effectively only by enzymatic hydrolysis or acid hydrolysis, lack of post-hypotonicity, post-fixation time shorter than 1 hour, and insufficient pressing force, etc., all of which affect the dispersion of cells and the separation of chromosomes

Method used

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  • Slide preparation method for discriminating chromosome number of Avena magna, Triticum aestivum or filial generation of Avena magna and Triticum aestivum
  • Slide preparation method for discriminating chromosome number of Avena magna, Triticum aestivum or filial generation of Avena magna and Triticum aestivum
  • Slide preparation method for discriminating chromosome number of Avena magna, Triticum aestivum or filial generation of Avena magna and Triticum aestivum

Examples

Experimental program
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Effect test

Embodiment 1 4

[0046]Example 1 Chromosome Number Analysis of 22 Strains of Tetraploid Large Oat and Common Wheat Hybrid Progeny

[0047] 1. Material preparation

[0048] Soak the seeds of 22 strains of tetraploid large oats and common wheat hybrid progenies obtained in water, place them on moist filter paper and cultivate them at 25°C for germination. When the roots grow to about 10mm, cut out the root tip meristem , quickly follow the steps below.

[0049] 2. Production

[0050] (1) Pretreatment: 0.075mol / L KCl solution, saturated α-bromonaphthalene solution, and distilled water are prepared as a pretreatment solution at a volume ratio of 2:3:5, and the cut root tips are soaked in the pretreatment solution, and treated at 25°C After 3 hours, wash with water for 15 minutes;

[0051] (2) Fixation: Methanol and glacial acetic acid were prepared into a fixative solution at a volume ratio of 3:1, and the pretreated root tips were fixed with the fixative solution, fixed at 25°C for 1 hour, and...

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Abstract

The invention discloses a slide preparation method for discriminating the chromosome number of Avena magna, Triticum aestivum or a filial generation of Avena magna and Triticum aestivum. The method comprises the following steps: pretreatment; fixation; enzymatic hydrolysis; acidolysis; post-hypotonic treatment; post-fixation; slide preparation; slide compression; etc. Through improvement of conventional methods, the slide preparation method provided by the invention can effectively discriminate the chromosome number of Avena magna, Triticum aestivum or the filial generation of Avena magna and Triticum aestivum. Experimental results show that a slide prepared by using the slide preparation method has a good number of cell division phases and good chromosome dispersion, and high repeatability and reliable results are obtained. Thus, a novel method is provided for discriminating the chromosome number of Avena magna, Triticum aestivum or the filial generation of Avena magna and Triticum aestivum, and technical support is provided for hybridization and seed selection of Avena magna and Triticum aestivum.

Description

technical field [0001] The invention relates to a method for preparing plant chromosomes, in particular to a method for effectively identifying the number of chromosomes of tetraploid large oats, common wheat or their hybrid offspring, and belongs to the field of biotechnology. Background technique [0002] Tetraploid large oat (Avenamagna L.) is a wild species, allotetraploid, showing high resistance or immunity to wheat powdery mildew, rust, scab, and has early maturity, drought resistance, protein (42%), High lysine content and other characteristics (Sun Zemin et al. Chinese oats. Beijing: China Agricultural Press, 1989; Yu Maoqun et al. Karyotype study of three oat species. Wuhan Botanical Research, 1995,13(2):177-179; Deng Guangbing et al. Oat interspecific hybrid F 1 Morphological and cytogenetic studies. Acta Crops, 2005,31(9):1186-1191.). In order to transfer these excellent traits of tetraploid oats to common wheat (Triticum aestivum L.), in the past ten years, ou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N1/30
CPCG01N1/28G01N1/30
Inventor 张飞雄杨才赵云云史文硕张彩云许梦月马克世
Owner CAPITAL NORMAL UNIVERSITY
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