A method for making slices for identifying chromosome numbers of tetraploid oats, common wheat or their hybrid offspring

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 (Avena magna L.) is a wild species, allotetraploid, showing high resistance or immunity to wheat powdery mildew, rust, scab, and early maturity, drought resistance, protein (42% ), high lysine content, etc. (Sun Zemin et al. Chinese oats. Beijing: China Agricultural Press, 1989; Yu Maoqun et al. Karyotype studies of three species of oats. 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.), our research group has carried out a ser...

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 0.075M KCl 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

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