高质量花生根尖细胞染色体制片方法研究

为了能快速获得用于染色体分带或者荧光原位杂交的高质量花生根尖细胞染色体制片,通过选择适当粗细的花生侧根,经过对二氯苯浸泡预处理,卡诺氏固定液固定后,直接用45%醋酸烘烤压片,得到了细胞平整、染色体分散的根尖细胞染色体制片。与HCl解离压片的方法相比,该方法中试剂对染色体影响微小。与酶解去壁低渗法相比,该方法没有酶解去壁低渗的过程,不需要用果胶酶和纤维素酶等较贵重试剂,节约了时间和成本,提高了制片效率。该方法不仅适用于花生,也适用于芝麻、水稻、玉米、大豆、青豆、番茄等多种植物。

Study on Slide Preparation Methods for High Quality Chromosomes from Root Tip Cells of Arachis

In order to get high quality slide preparation to meet the requirements of chromosome banding and in situ hybridization,the cytogenetic slide preparation with evenly dispersing celllayers and well spreading chromosomes was obtained from root tip cells of Arachis.The key procedures included selecting of roots with appropriate size,pretreating with p-dichlorobenzene and fixing with Carnoy's solution,and making slide directly after heating in 45% glacial acid.Thismethod had little effect on chromosome compared with the method of HCl disintegrating.By saving the costly pectinase and cellulose through omitting steps of enzymatic hydrolysis and hypotonic to cell wall,the method saved time and costs.This method was also applied to chromosome preparation of other crops such as sesame,rice,corn,soybean,green bean and tomato,which again proved to be efficient.