Role of polyamines in the cytokinin-dependent physiological processes. I. Effect of benzyladenine on polyamine levels during chloroplast differentiation in the tissue culture of Dianthus caryophyllus

Abstract

Tissue culture of Dianthus caryophyllus L. (cv. William Sire.) obligatory requiring N6-bermyladenine for greening provides a good system to study the interactions between cytokinins and polyamines. Polyamines were analyzed as dansyl derivatives which are separated by thin layer chromatography and detected by fluorescence spectrophotometry.Green callus growing on benzyladenine -containing medium showed decrease in the contents of free, conjugated and bound putrescine and spermidine in comparison to chlorophyll-less callus (control callus) growing on cytokinin-free medium. The level of spermine free, conjugated and bound forms increased about 6 %, 77 % and 28 % respectively in tissue culture growing in the presence of cytokinin.Spermidine was dominant polyamine bound to ehromatin isolated from control callus. Chromatin isolated from green-callus was characterized by a lower level of each polyamine in comparison to chlorophyll-less callus.Polyamines were found in plastid membrane fraction isolated from chlorophyll-less and green callus. A significant increase the levels of polyamines (putrescine, spermidine and spermine) bound to plastid membranes in green callus (+ benzyladenine) in comparison to chlorophyll-less callus (-benzyladenine) was observed. Additionaly, methylglyoxal-bis(guanylhydrazone) an inhibitor of S-adenosylmethionine decarboxylase depressed the greening process.Our results suggest that cytokinin-induced chloroplast differentiation in carnation tissue culture may be partly mediated through the polyamines bound to thylakoid membranes. A possible role of polyamines during cytokinin-induced formation of photosynthetic apparatus is discussed.

Polyamine metabolism and S-adenosylmethionine decarboxylase gene expression during the cytokinin-stimulated greening process

“…It has been shown that in green tissue culture of Dianthus caryophyllus L. (cv. William Sim), which required N 6 -benzyladenine for greening, a significant increase in the level of Put, Spd and Spm bound to plastid membranes was observed in comparison to the control (chlorophyll-less callus) (Legocka and _ Zarnowska 1999). Kotzabasis et al (1993a, b) and Del Duca et al (1994) reported that the main polyamines: Put, Spd, Spm are associated with the light-harvesting complex (LHC) and the photosystem II (PSII) of spinach and Helianthus tuberosus.…”

“…As suggested by Legocka and _ Zarnowska (1999 polyamines might be involved in cytokinin-mediated mechanisms of chloroplast differentiation in tissue culture of carnation, expansion of cucumber cotyledons or gametophore bud formation of moss. It has been shown that in green tissue culture of Dianthus caryophyllus L. (cv.…”

Action of humic acid on promotion of cucumber shoot growth involves nitrate-related changes associated with the root-to-shoot distribution of cytokinins, polyamines and mineral nutrients

“…These effects on PA distribution within the plant could be linked to those related to CKs. In fact, some studies have reported that CK application caused significant increases in put concentration in excised cotyledons (Legocka and Zarnowska, 1999;2000;Sobieszczuk-Nowicka et al, 2007). These studies showed that this increase in put was not essential for the CK-induced expansion of cotyledons (Legocka and Zarnowska, 2000).…”

Plastid‐membrane‐associated polyamines and thylakoid transglutaminases during etioplast‐to‐chloroplast transformation stimulated by kinetin

“…cotyledons (Cho 1983), C. sativum cotyledons (Legocka and Żarnowska 2000, Suresh et al 1978, Walker et al 1988) and C. purpureus tissue culture (Legocka and Żarnowska 2002). Furthermore, our previous study (Legocka and Żarnowska 1999) suggested that cytokinin‐induced chloroplast differentiation in carnation tissue may be partly mediated by PAs bound to thylakoid membranes, but the molecular mechanism still needs to be clarified.…”