Orcein staining and the study of the effect of chemicals on chromosomes

Summary

Plant materials of both dicotyledonous and monocotyledonous groups were subjected to treatment with alloxan followed by coumarin, phenols, oxyquinoline etc. for 3 to 4 hours.

Root tips were then heated in an orcein-acid mixture for 10 to 40 seconds and studied after mounting in 1% orcein. Clear metaphase plates were obtained after 10 seconds of heating, whereas after 20 and 30 seconds considerable erosion and fragmentation respectively were recorded. Complete loss of stainability resulted after heating for 40 seconds.

Fragmentation is caused only by orcein under heat conditions. This is demonstrated by control series checking all variables.

Pretreatment with the chemicals stated is of absolute necessity for fragmentation. It is suggested that this pretreatment causes depolymerisation of DNA and lability of nucleoprotein linkage at certain segments from which the DNA becomes detached during subsequent heating.

Of all the chemicals tested for pretreatment effects, marked positive results were obtained with alloxan (1 hour) followed by coumarin (2 hours at 16–20 ° C.). Phenols also gave positive results. The other chemicals tried need slightly longer time of treatment for fragment induction.