Petr GALUSZKA Centre of the Region Haná for Biotechnological and Agricultural Research Palacký University in Olomouc FROM CYTOKININ METABOLISM TO AGRICULTURE.

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Petr GALUSZKA Centre of the Region Haná for Biotechnological and Agricultural Research Palacký University in Olomouc FROM CYTOKININ METABOLISM TO AGRICULTURE

Gajdošová et al., JXB 2011 CK qualitative content analysis barleymaizeCK-typeArabidopsis pmol/g of leaf fresh weight iP tZ cZ DHZ4.2

CK qualitative analysis during maize development pmol/g of leaf fresh weight kernel germination shoot developmentroot development 20th day 20th day isopentenyladenine trans-zeatin cis-zeatin dihydrozeatin

CYTOKININ SENSING in Zea mays Histidine kinase – CK receptor bound to membranes Apparent K D (nM) CKZmHK1ZmHK2ZmHK3 isopentenyladenine trans-zeatin cis-zeatin dihydrozeatin tZR iPRnot detectable400 tZOGnot detectable S. Lomin et al., JXB 2011 Contrary to Arabidopsis receptors, there is higher sensitivity to cZ and DHZ at least on some maize receptors

iP ? cZ DHZ tZ zeatin metabolism in planta is rather obscure de novo biosynthesis tRNA degradation CYP450 zeatin isomerase IPT LOG zeatin reductase ? proved in planta gene is known proved in vitro activity detected hypothesized

Cytokinin dehydrogenases have different preferences CKXiPtZcZDHZ AtCKX AtCKX AtCKX AtCKX AtCKX AtCKX AtCKX ZmCKX ZmCKX HvCKX CKX relative activity CKX gene families: Arabidopsis thaliana7 genes Zea mays13 genes Oryza sativa11 genes Hordeum vulgare11 genes

CKX as a transgene Constitutive overexpression of CKX causes cytokinin-deficient phenotype Main function of CKs on plant differentiation proved together with opposite role on root and shoot development Cytokinin content 30–50% of WT Construct: 35S::HvCKX2 pmol/g fresh w CKWTHvCKX2 seedlings iP tZ

Transformation of Barley with CKX genes Ubiqutin::ZmCKX1 Ubiqutin::HvCKX2 WT trsg WT Z2 Z4 Z7 Z8 Z10 WTZ10 Constitutive CKX overexpression in cereal: prematured senescence no onset of flowering (sterility) enlarged root system (dependent of transgene) pmol/g fresh w CKWTtrsg root iP tZ cZ DHZ shoot iP tZ cZ DHZ2.62.9

CKX and IPT gene families in barley 11 CKX genes 2 tRNA IPT genes 5 de novo IPT genes

wild type situation our transgenic approach