Calcium‐calmodulin in germination of Phacelia tanacetifolia seeds: effects of light, temperature, fusicoccin and calcium‐calmodulin antagonists

Abstract

Germination in the dark and at 16°C of photoblastic and thermosensitive seeds of Phacelia tanacetifolia was inhibited when incubated with EGTA and the Ca2+‐ionophore A 23187; A 23187 in the presence of Ca2+ still inhibited germination, but to a lesser extent. Treatments with EGTA or Ca2+ at different concentrations in the presence or in the absence of A 23187 did not remove light inhibition. The calmodulin (CaM) inhibitor, calmidazolium, strongly inhibited germination. The specificity of these inhibitors and their effects on seed germination are discussed.CaM from Phacelia tanacetifolia seeds has been purified and its characteristics (molecular weight, heat and acid stability, kinetics of phosphodiesterase [EC 3.1.4.17] activation) were very similar to those of other plant sources. More than 90% of total CaM was present in the soluble fraction (ca 41 μg g‐1 fresh weight in ungerminated seeds). The CaM level greatly increased in the early phases of seed germination; this increase did not take place when germination was inhibited by light or high temperature. When fusicoccin, a toxin which promotes germination by activating membrane functions, relieved light or high temperature inhibition, CaM increased up to the control value in the dark at 16°C. The parallel increase in CaM and seed germination suggest that CaM plays an important role in the process. Fusicoccin in the dark at 16°C stimulated CaM and fresh weight increase, but not the metabolic reactivation measured as increase in DNA and total RNA levels; at 30°C fusicoccin stimulated the increase in fresh weight and in CaM level, but the increases in DNA and total RNA were very low. These results suggest that the activation of membrane functions with cell enlargement induced by fusicoccin is related to CaM increase.