生物技术通报2016，Vol.32Issue(4)：74-79,6.DOI:10.13560/j.cnki.biotech.bull.1985.2016.04.009
莲藕根尖染色体制片与免疫荧光染色技术的优化
The Optimization of Chromosome Preparation and Immunofluorescence Staining for Root Tip of Nelumbo nucifera
摘要
Abstract
It is difficult to prepare lotus’s(Nelumbo nucifera)root tip chromosome in the cytogenetic study. Therefore,some related factors were studied,such as selecting the proper length of root tip,mitotic index,pretreatment medicament and time,the fixing time,the time of enzyme hydrolysis,and low osmosis in the chromosome preparation from the root tip cells of lotus,and then the immunofluorescence staining was also carried out to validate the chromosome preparation. The results showed that the mitotic index was the highest and morphology was clear when the root tips length was approximately 1.5 cm and the root tips was treated with ice water for 24 hours. The chromosome morphology was solid while root tips was for 1 h fixing with 4% paraformaldehyde. The enzyme hydrolysis was complete,the residue of cytoplasm was little,and the chromosome scattered well while the tips were digesting 30 min with combination of 2.5%cellulase and 2.5%pectinase at 37℃. The chromosome morphology was fine under 30 min low osmosis treatment by 1×PBS. Strong signals were detected after the immunofluorescence staining. This study set up the foundation for the relevant researches on cytogenetic and epigenetic studies of N. nucifera.关键词
莲藕根尖细胞/预处理/有丝分裂指数/染色体制片/DAPI染色/免疫荧光Key words
root tip cells of Nelumbo nucifera/pretreatment/mitotic index/chromosome preparations/DAPI staining/immunofluorescence引用本文复制引用
王海燕,朱之轩,金晶,丁毅..莲藕根尖染色体制片与免疫荧光染色技术的优化[J].生物技术通报,2016,32(4):74-79,6.基金项目
国家自然科学基金资助项目(31271310) （31271310）
