高质量杨树粗线期染色体制片及荧光原位杂交  

The High Quality Slides of Pachytene Chromosome Preparation and Fluorescence in situ Hybridization in Populus deltoides

作　　者：辛昊阳 兰月[1] 施季森[1] 戴兆霞 席梦利[1] 

机构地区：[1]南京林业大学南方现代林业协同创新中心,南京210037

出　　处：《分子植物育种》2016年第3期655-659,共5页Molecular Plant Breeding

基　　金：江苏高校优势学科建设工程项目资助

摘　　要：以美洲黑杨无性系‘哈佛杨’处于减数分裂粗线期的花药为研究材料,比较了蒸汽滴片法、涂片法、压片法及改良压片法4种不同制片方法,在制备美洲黑杨粗线期染色体中的差异,以期获得高质量的美洲黑杨粗线期染色体制片。结果表明:改良压片法是获得高质量美洲黑杨粗线期染色体的有效方法,获得的染色体制片背景干净,染色体形态良好,适于进一步荧光原位杂交研究。以拟南芥端粒序列为探针,采用缺刻平移法用地高辛标记探针DNA,杂交后信号用Rodamine anit-DIG-sheep抗体检测,建立了高效的荧光原位杂交技术体系。为进一步开展杨树的分子细胞遗传学研究奠定基础。In this research,anthe rs at the stage of pachytene in Populus deltoides Marsh were used as materials.Four different methods('steamdrop'method,smear method,squash method,modified squash method) were compared to obtain the high quality slides of pachytene chromosomes.The results showed that modified squash method is an effective method to obtain the high quality slides of pachytene chromosomes with a clear background and well shaped pachytene chromosomes.Therefore,the high quality slides of pachytene chromosomes can be suitable for fluorescence in situ hybridization further more.Finally,a highly efficient fluorescence in situ hybridization technique system was established with a probe of Arabidopsis telomere sequence,the probe was labeled with digoxigenin-11-d UTP using nick translation,and detected using Rodamine anit-DIG-sheep.The research was to lay the foundation for improving the molecular cytogenetic study of Populus.

关 键 词：美洲黑杨 粗线期染色体 荧光原位杂交 

分 类 号：S792.11[农业科学—林木遗传育种]