用于流式细胞术分析的杉木根尖细胞同步化及染色体分离
Cell synchronization and isolation of chromosomes from Chinese fir root tips for flow cytometric analysis.
作者信息
Shen Jie, Xu Jin, Chen Jinhui, Zheng Renhua, Shi Jisen
机构信息
Key Laboratory of Forest Genetics and Biotechnology, Ministry of Education, Nanjing Forestry University, Nanjing, 210037, China.
出版信息
Biotechnol Lett. 2015 Jun;37(6):1309-14. doi: 10.1007/s10529-015-1800-x. Epub 2015 May 8.
OBJECTIVES
To optimize treatment parameters to synchronize the cell cycle of Chinese fir root tip meristem cells.
RESULTS
Optimal mitotic indices were achieved by treating seedlings with 1.25 mM hydroxyurea for 10 h, incubating for 7 h without chemical treatment to release the hydroxyurea block, and then treating emerging roots with 0.05 % colchicine for 16 h. The mitotic index of synchronized Chinese fir root tips was over 30 %. Suspensions of intact chromosomes were prepared using a simple slicing procedure.
CONCLUSIONS
The chromosome preparations were suitable for flow cytometric characterization and sorting. The chromosome peaks of the observed flow karyotype resembled the predicted flow karyotype calculated from the Chinese fir chromosome size.
目的
优化处理参数以同步杉木根尖分生组织细胞的细胞周期。
结果
通过用1.25 mM羟基脲处理幼苗10小时，在无化学处理的情况下孵育7小时以解除羟基脲阻断，然后用0.05%秋水仙碱处理新生根16小时，获得了最佳有丝分裂指数。同步化杉木根尖的有丝分裂指数超过30%。使用简单的切片程序制备了完整染色体的悬浮液。
结论
染色体标本适用于流式细胞术表征和分选。观察到的流式核型的染色体峰与根据杉木染色体大小计算出的预测流式核型相似。
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