甘蔗根尖染色体制片技术研究

Studies on Technique of Chromosome Squashing of Root-tip in Sugarcane

甘蔗是异源多倍体植物,染色体数目多、体积小,染色体制片困难,导致细胞遗传学研究进展缓慢。以分裂旺盛的甘蔗根尖为材料,对不同采样时间、预处理方法、解离时间、染色方法等各技术环节进行实验研究。结果表明:在冬季气温较低的条件下,于15:00进行采样,饱和对二氯苯与0.002mol/L8-羟基喹啉等体积混合液室温预处理4～4.5h,1mol/LHCl与45%乙酸等体积混合液于60℃水浴锅解离8min后,用改良苯酚卡宝品红进行涂片滴染,所获得的染色体制片效果最好。

Sugarcane belongs to the allopolyploid plant which contains large number of chromosome with small size.The cytological research on sugarcane may hence be influenced by its difficulties to obtain chromosome products successful.In this paper,the root-tips of sugarcane were used to study the effects of the sampling time,the pretreatment chemical solution,hydrolysing time and staining on the chromosome squashing.The results showed that the optimal sampling time of roots was 15：00 pm under the lower temperature condition in winter.Among the four pretreatment of chemical solutions,the solution mixed with equal volume of saturated p-dichlorobenzene and 0.002 mol/L 8-hydroxyquinoline,was pretreated at room temperature for 4-4.5 h got best result.Root tips were then hydrolysed with the solution mixed by equal volume of 1 mol/L HCl and 45% acetic acid,for 8 min at 60℃,and stained with modified Carbol fuchsin solution.With this method a very good effect of chromosome squashing could be obtained.