摘要
[目的]探索速生杨愈伤组织染色体制片的可行措施。[方法]以中林46号速生杨愈伤组织为材料,取2 mm^3左右生长旺盛的愈伤组织块,采用不同的预处理方法、解离及染色方法制备永久封片,对比不同方法的制片效果。[结果]在5种预处理方法中,愈伤组织用4℃冰水预处理24 h或用0.05%秋水仙素在8-16℃条件下预处理6 h时,制片效果较好;在2种解离方法中,酸解和酶解愈伤组织效果相当,较理想;在3种染色方法中愈伤组织用吉姆萨染色2 h左右效果较好,改良苯酚品红染色2 h效果次之,醋酸洋红染色较浅,其显微摄影效果不理想。[结论]用4℃冰水或0.05%秋水仙素预处理、用盐酸或含果胶酶、纤维素酶的甘露醇溶液解离、用吉姆萨染色时,速生杨愈伤组织染色体制片效果较好。
[ Objective] The study was to explore the feasible measures on chromosome preparation of the callus of fast-growing poplar. [ Method] With callus of fast-growing poplar of Zhonglin 46 as material, the eugonic callus with about 2 mm^3 were taken to prepare the permanent chromosome preparation with different methods of pretreatment, dissociation and staining and their the preparation effects were compared. [ Result] Among 5 pretreatment methods, the preparation effects were better when the callus was pretreated by ice water at 4 ℃ for 24 h or pretreated by 0.05% colchicine at 8 - 16 ℃ for 6 h. Awing 2 dissociation methods, the effects of acidolysis and enzymatic hydrolysis on callus were equivalent, and they all were more perfect. Among 3 staining methods, the staining results of Giemsa was better when the callus was stained for about 2 h, the second was that stained by improved phenol fuchsin for about 2 h, and the staining color by aceto carmine was lighter, so its effect of photomicrography was non-ideal. [Conclusion] The chromosome preparation effect of fast-growing poplar callus was better when callus was pretreated by ice water at 4 ℃ or by 0.05% colchieine, and dissociated by hydroehloride or mannitol solution that contained pectinase and eellulase and stained by Giemsa.
出处
《安徽农业科学》
CAS
北大核心
2008年第25期10895-10896,10907,共3页
Journal of Anhui Agricultural Sciences
基金
新疆生物资源基因工程重点实验室资助项目
关键词
速生杨
染色体
愈伤组织
Fast-growing poplar
Chromosome
Callus