Acta Univ. Agric. Silvic. Mendelianae Brun. 2010, 58(4), 79-88 | DOI: 10.11118/actaun201058040079

Cytokininy a polární transport auxinu v axilárních pupenech hrachu

Petr Kalousek, Dagmar Buchtová, Jozef Balla, Vilém Reinöhl, Stanislav Procházka
Ústav biologie rostlin, Mendelova univerzita v Brně, Zemědělská 1, 613 00 Brno, Česká republika

V této práci byl sledován vliv cytokininu na polární transport auxinu v axilárních pupenech 2 nodu sedmidenních rostlin hrachu setého (Pisum sativum L.) cv. Vladan v průběhu jejich uvolňování z apikální dominance. V axilárních pupenech dekapitovaných, příp. exogenní aplikací benzyladeninu ošetřených intaktních rostlin byla metodou dvoukrokové, semikvantitativní RT PCR studována exprese genů PsAUX1 (AUXIN RESISTANT 1) a PsPIN1 (PIN-FORMED 1), které kódují auxinové přenašeče, a genů PsDRM1 (DORMANCY-ASSOCIATED PROTEIN 1) a PsAD1 (APICAL DOMINANCE 1), jejichž exprese souvisí s dormantním, resp. korelačně inhibovaným stavem pupenů. Pro srovnání byla sledována i exprese těchto genů v axilárních pupenech ošetřených exogenně aplikovaným auxinem. Exprese genu PsDEADbox byla použita jako vnitřní kontrola. Lokalizace PIN1 proteinu, klíčového faktoru polárního transportu auxinu v axilárních pupenech, byla vizualizována imunohistochemicky na podélných řezech axilárním pupenem podle metodiky Paciorek et al. (2006). Objekty byly snímány laserovým konfokálním mikroskopem TCS SP2 AOBS (Leica, Mannheim, Germany). Po exogenní aplikaci cytokininu na axiláry byl prokázán rychlý vzestup exprese genů PsAUX1 a PsPIN1 se současným poklesem exprese genů PsDRM1 a PsAD1. Obdobné změny v expresi byly zaznamenány rovněž v axilárech dekapitovaných rostlin, byly však zřetelně pomalejší. Auxinový přenašeč PsPIN1 protein nebyl v axilárních pupenech intaktních rostlin lokalizován polárně. Po aplikaci cytokininu došlo k postupné polarizaci PIN1 proteinu na bázi auxin transportujících buněk. Přestože přímá aplikace auxinu na axilární pupeny intaktních rostlin vedla rovněž ke zvýšení exprese genů PsAUX1 a PsPIN1, pupeny zůstaly inhibovány, což prokazuje přetrvávající exprese markerů dormantního stavu genů PsDRM1 and PsAD1. Dosažené výsledky ukazují na možný efekt cytokininů na biosyntézu, respektive transport auxinu v axilárních pupenech a vyzdvihují význam vzájemných interakcí auxinu a cytokininů při uvolňování pupenů z lodyžní apikální dominance.

apikální dominance, hrách setý, cytokininy, polární transport auxinu

Cytokinins and polar transport of auxin in axillary pea buds

The influence of cytokinin on auxin transport during release of axillary buds from apical dominance was studied. Expression of auxin-carrier coding genes PsAUX1 (AUXIN RESISTANT 1) and PsPIN1 (PIN-FORMED 1) was explored in axillary buds of the 2nd node of 7-day pea plants (Pisum sativum L.) cv. Vladan after decapitation or after exogenous application of benzyladenine (6-benzylaminopurine) onto axillary buds of intact plants. Localization of the PsPIN1 protein, the key factor for polar transport of auxin in axillary buds, was visualised by immunohistochemistry. After exogenous application of cytokinin the expression of PsAUX1 and PsPIN1 rapidly increased with a simultaneous rapid decrease in PsDRM1 and PsAD1 expression - genes related to bud dormancy. The same changes in expression were observed after decapitation, however they were markedly slower. The PsPIN1 auxin efflux carrier in the inhibited axillary buds of intact plants was localised in a non-polar manner. After exogenous application of cytokinin gradual polarisation of the PsPIN1 protein occurred on the basal pole of polar auxin transport competent cells. Despite the fact that direct auxin application to buds of intact plants led to an increase in PsAUX1 and PsPIN1 expression, the buds remained dormant (non-growing) what was accompanied by persistent expression of the dormancy markers PsDRM1 and PsAD1. The results indicate a possible effect of cytokinins on biosynthesis, and/or transport of auxin in axillary buds and they highlight the importance of auxin-cytokinin crosstalk in the regulation of bud outgrowth after breaking of apical dominance.

Keywords: apical dominance, cytokinins, pea, polar auxin transport
Grants and funding:

This work was supported by a grant of the Ministry of Education of the Czech Republic No. LC06034.

Received: May 31, 2010; Published: September 15, 2014  Show citation

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Kalousek, P., Buchtová, D., Balla, J., Reinöhl, V., & Procházka, S. (2010). Cytokinins and polar transport of auxin in axillary pea buds. Acta Universitatis Agriculturae et Silviculturae Mendelianae Brunensis, 58(4), 79-88. doi: 10.11118/actaun201058040079
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