A Root Squash Technique for Counting Somatic Chromosomes in Sugar Cane

Autor: N. S. Sisodia
Rok vydání: 1968
Předmět:
Zdroj: Stain Technology. 43:129-134
ISSN: 0038-9153
DOI: 10.3109/10520296809115055
Popis: Sugar cane cuttings after treatment with 0.75% Aeratan (organic mercurial fungicide) were germinated in a soil mixture at 26–28 C and transferred to an incubator at 33–34 C for 1 day before collecting roots. The excised roots were pretreated with a saturated solution of α-bromonaphthalene in 0.05% saponin for 3 hr at 8–10 C, fixed in 1:3 acetic-alcohol for 48–72 hr at room temperature, hydrolysed in 1 N HCl for 7–8 min at 60 C, treated with 3% pectinase solution in pH 3.6 acetate buffer for 60–90 min, stained by the Feulgen procedure, and squashed in 1% acetocarmine. Following this procedure, excellent preparations were obtained for counting chromosomes, in which the details of chromosome morphology were often clearly visible. The important precautions for the successful application of this technique include: avoidance of excessive watering of the germinating cuttings, avoidance of sudden temperature changes during processing of unfixed roots, ensuring correct acid temperature during hydrolysis and, final...
Databáze: OpenAIRE