Publication details

Esterases as a marker for growth of BY-2 tobacco cells and early somatic embryos of the Norway spruce.

Authors

VÍTEČEK Jan ADAM Vojtěch PETŘEK Jiří VACEK Jan KIZEK René HAVEL Ladislav

Year of publication 2004
Type Article in Periodical
Magazine / Source Plant cell tissue and organ culture
MU Faculty or unit

Faculty of Science

Citation
Field Physiology
Keywords BY-2; cell viability; early somatic embryo; esterase; fluorescein diacetate; Nicotiana tabacum; Picea abies
Description Growth is one of the basic properties of biological systems. The methods which are commonly used for the determination of growth are usually difficult and not very accurate. In the present work we decided to use esterase activity as a growth marker in tobacco suspension culture (BY-2 line) and in early somatic embryos of Norway spruce (clone 2/32) grown on a semi-solid medium. Esterase activity correlates well with the classical growth characteristics of BY-2 and spruce early somatic embryos. Determination of esterase activity is based on spectrophotometric and spectrofluorimetric detection of reaction products, which arise from the enzymatic hydrolysis of two substrates (p-nitrophenyl acetate and fluorescein diacetate) by esterase. The spectrophotometric method enabled us to detect approximately 10(4) BY-2 cells and 25 spruce embryos whereas the more sensitive spectrofluorimetric method allowed us to detect approximately 800 BY-2 cells and 5 early somatic embryos of Norway spruce.

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