Calcium Variously Mediates the Effect of Cytokinin on Chlorophyll and LHCPII Accumulation During Greening in Barley Leaves and Cucumber Cotyledons

During greening, excised etiolated barley leaves and cucumber cotyledons that were depleted of exogenous Ca2+ by a chelating agent (ethylene glycol-bis (beta aminoethyl ether)-N,N,N`N`-tetraacetic acid, EGTA) showed ∼50% reduced chlorophyll (Chl) accumulation and ∼30% accumulation of apoprotein of the light-harvesting chlorophyll a/b-binding protein complex of photosystem II (LHCPII). The Ca2+ channel blocker lanthanum chloride (LaCl3) applied to cucumber cotyledons reduced LHCPII accumulation more than EGTA did. In both plant mate-rials, cytokinins enhanced chlorophyll accumulation by 50-60% and this effect was completely canceled by EGTA application. Hormones significantly increased LHCPII accumulation but EGTA application reduced that effect in barley leaves by ∼30% and in cucumber cotyledons by ∼80%. A similar effect was observed in LaCl3-treated cotyledons. CaCl2 application boosted chlorophyll accumulation in both plant materials. CaCl2 applied together with cytokinin reduced the hormonal effect on chlorophyll accumulation by ∼38% in barley leaves and 23% in cucumber cotyledons, but almost totally inhibited cytokinin-stimulated LHCPII accumulation. Our results indicate that calcium variously mediates the effect of cytokinin on chlorophyll and LHCPII accumulation. Cytokinin-induced enhancement of chlorophyll accumulation seems totally dependent on the exogenous pool of Ca2+, while Ca2+-dependent and Ca2+-independent pathways are involved in the hormonal effect on LHCPII accumulation. The effect of cytokinin on the increase of light-induced LHCPII accumulation appears to be sensitive to exogenously applied Ca2+, which almost totally blocked the hormonal effect. Our results give indirect evidence that the responses to cytokinin and light act on different events leading to Chl and LHCPII accumulation.

No effect of plant growth retarding compounds and growth stimulators on indolo-3-acetic acid oxidase activity in greening cucumber cotyledons

Cotyledons dissected from 5-day-old etiolated cucumber seedlings were incubated in solutions on AMO-1618, B-Nine, CCC and Phosfon D for 48 h in light. In some tests the retardants were applied in mixed solutions with GA₃ or BAP. IAA oxidase was extracted and purified by means of molecular sieving through a bed of Sephadex G-25. The retardants inhibited chlorophyll synthesis by 50 % or more, and had essentially no effect on IAA oxidase activity per cotyledon basis. GA₃ and BAP also had no effect on enzyme activity in spite of a fact that the compounds stimulated growth of the cotyledons. The crude enzyme extract from B-Nine treated cotyledons showed lower IAA oxidase activity in comparison with the water treated control, the effect being due to a longer lag-phase preceding the initiation of IAA oxidation. KNO₃ strikingly stimulated expansional growth of the cotyledons, the effect being correlated with the accelerated chlorophyll accumulation. KNO₃ had no effect on IAA oxidase activity per cotyledon and decreased it per gram fr wt. It is concluded that [1] the growth rate of cucumber cotyledons is not correlated with IAA oxidase activity, and ;[2] the growth retarding compounds do not affect IAA oxidase system is this tissue.

Hamowanie syntezy chlorofilu i RNA w izolowanych liścieniach ogórka przez N-hydroksymocznik [Inhibition of chlorophyll and RNA synthesis by N-hydroxyurea in detached cucumber cotyledons]

N-hydroxyurea (HU) at the concentration of 5 x 10^-4 M decreased the content of chlorophyll in detached cucumber cotyledons; at this concentration it has no inhibitory effect on growth. Benzylaminopurine, gibberelic acid and KCl partially reversed the inhibitory effect of HU on chlorophyll synthesis. HU stimulated yellowing of barley first leaf sections. The compound had little effect on leucine-¹⁴C incorporation to protein, and markedly inhibited uracil-¹⁴C incorporation in to RNA of the greening cucumber cotyledons. It is suggested that the inhibition of RNA and chlorophyll synthesis in HU-treated cucumber cotyledons follows the HU-dependent inhibition of DNA replication.

Effect of 2-chloroethylphosphonic acid and vanillin on chlorophyll, protein and RNA synthesis in detached cucumber cotyledons, and chlorophyll degradation in senescing leaf discs of kale

N⁶-benzylaminopurine (BAP) and KC1 stimulated growth in detached greening cucumber cotyledons; BAP enhanced RNA synthesis whereas KCl strikingly accelerated leucine-¹⁴C incorporation into proteins. Vanillin (0.01 M) inhibited greening and protein synthesis, and nullified the stimulatory effects of KC1. a-chloroethylphosphonic acid (CEPA, 0.01 M) inhibited greening without any effect on protein synthesis when it was applied alone; CEPA decreased RNA synthesis, completely nullified the K⁺-induced synthesis of chlorophyll, and reduced the BAP^- or KCl^-induced growth. Vanillin inhibited yellowing in 'senescing leaf discs of kale kept in darkness, whereas CEPA had an opposite effect.