滇杨根尖细胞染色体制片技术优化  

Technique optimization on chromosome preparation of Populus yunnanensis Dode root-tip cell

作　　者：尹丽莎 陈杰 周军[1,2] 何承忠[1,2] 李斌[1] 张辉[1] 罗一然[1] 辛培尧[1,2] 

机构地区：[1]西南林业大学/云南省高校林木遗传改良与繁育重点实验室,昆明650224 [2]西南林业大学林学院,昆明650224 [3]毕节市科技局中药研究所,贵州毕节551700

出　　处：《南方农业学报》2015年第7期1253-1258,共6页Journal of Southern Agriculture

基　　金：国家林业局林业公益性行业科研专项项目(201104076);云南省省院省校教育合作咨询共建重点学科项目(211015);云南省教育厅科学研究基金重大专项项目(ZD2015-014)

摘　　要：【目的】获取滇杨根尖细胞染色体优良制片,为滇杨染色体数目鉴定及遗传改良提供参考依据。【方法】以滇杨试管苗根尖为材料,采用常规制片法制片,筛选出获取最佳染色体制片效果的预处理液、预处理时间、固定时间、解离液种类及解离时间、染色时间等处理条件,并鉴定滇杨染色体数目。【结果】滇杨根尖材料在0℃饱和对二氯苯溶液中预处理1.0 h,再转入0℃卡诺固定液中固定1.0 h,随后在浓盐酸∶无水乙醇=1∶1的解离液中常温解离4.0min,常规压片法染色、压片,其染色体制片着色良好、清晰、无杂质。经显微观察,确定滇杨染色体数目为2n=2X=38。【结论】在建立的滇杨根尖染色体制片技术体系下,可获得良好的染色体制片效果,快速清晰观察到滇杨的染色体数目。[Objective]The present experiment was conducted to investigate the optimal chromosome preparation tech- nique of P. yunnanensis Dode root-tip cell, in order to provide references for identification of P. yunnanensis Dode＇s chromosome number and its genetic improvement. [Method]Taking test-tube plantlet root tip of P.yunnanensis Dode as materials, the conventional chromosome preparation method was used to optimize pretreatment solution,pretreatment time, fixing time, type of dissociated solution, dissociated time and dyeing time, in order to obtain optimal chromosome prepara- tion effect. Besides, the chromosome number of P. yunnaneusis Dode was identified. [Result]After being pretreated in sat- urated p-dichlorobenzene solution at 0 ℃ for 1.0 h,the root-tip materials were fixed in Carnot solution for 1.0 h at 0 ℃. Then, these materials were dissociated in mixed solution （concentrated hydrochloric acid：anhydrous ethanol=1：1）for 4 min at room temperature. In addition, after being dyed in modified Carbol fuchsin solution for 9 min, the clear and favor- able chromosome images with good dyeing effect were obtained with traditional squash methods. Furthermore, the chromo- some number of P. yunuanensis Dode was 2n=2X=38 through observing with microscope. [Conclusion ]The established root-tip chromosome preparation technique system of P. yunnanensis Dode can make root-tip chromosome number be observed quickly and clearly, so as to obtain favorable chromosome images with good dyeing effect.

关 键 词：滇杨 染色体制片 染色体数目鉴定 

分 类 号：S792.118[农业科学—林木遗传育种]