南方农业学报2017，Vol.48Issue(4)：663-668,6.DOI:10.3969/j.issn.2095-1191.2017.04.016
多倍体西瓜染色体制片技术改良及其核型分析
Optimization of chromosome preparation and karyotype analysis of polyploid watermelons
摘要
Abstract
[Objective]Technique of chromosome preparation for polyploid watermelon was improved,conventional karyotype and fluorescence karyotype for polyploid watermelons were compared in order to provide reference for research on genetics of watermelon and establishment of physical genetic map of watermelon . [Method]Polyploid watermelons were used as materials in the study. Based on traditional chromosome preparation technique , namely wall degradation-hypotonic flaming-drying method, pre-hypotonic and after-hypotonic were deleted, and re-fixation and flame smear were combined into one procedure. Giemsa stain and DAPI fluorescence stain were used to stain, and carried out conventional karyotype analysis and fluorescence karyotype analysis respectively. [Result]The optimized enzymolysis flaming-drying method was used to prepare chromosome of polyploid watermelons. Compared with the traditional technique , the optimized one simplified three procedures which were pre-hypotonic and after-hypotonic of KCL solution and re-fixation by Carnoy fixative. Thus the time was 1.0-2.0 h shorter. Production efficiency was improved, and samples with complete amount, good dispersion and clear configuration were obtained. Slides stained with DAPI could shorten the detection time for 0.5-1.5 h each slide, and were more easily to be detected metaphases with more clear centromere,long arm and short arm of chromosome than stained with Giemsa . [Conclusion]The modified method can improve the efficiency of chromosome preparation of polyploid watermelon. Fluorescent karyotype analysis is more accurate and efficient than conventional karyotype analysis.关键词
多倍体西瓜/染色体制片/荧光染色/核型分析Key words
polyploid watermelon/chromosome preparation/fluorescence stain/karyotype analysis分类
农业科技引用本文复制引用
李桂芬,李文信,何毅,覃斯华,黄金艳,柳唐镜,洪日新,李天艳,樊学军,韦正光..多倍体西瓜染色体制片技术改良及其核型分析[J].南方农业学报,2017,48(4):663-668,6.基金项目
国家西甜瓜产业技术体系建设项目(CARS-26) （CARS-26）
广西自然科学基金项目(2013GXNSFBA019099) （2013GXNSFBA019099）
广西农业科学院科技发展基金项目(桂农科2013JZ02,桂农科2015JZ013) （桂农科2013JZ02,桂农科2015JZ013）
