Expression of cytokinin biosynthetic isopentenyltransferase genes in <i>Arabidopsis</i>: tissue specificity and regulation by auxin, cytokinin, and nitrate

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<jats:title>Summary</jats:title><jats:p>The rate‐limiting step of cytokinin biosynthesis in <jats:italic>Arabidopsis thaliana</jats:italic> Heynh. is catalyzed by ATP/ADP isopentenyltransferases, <jats:italic>A. thaliana</jats:italic> IsoPentenyl Transferase (AtIPT)1, and AtIPT4, and by their homologs AtIPT3, AtIPT5, AtIPT6, AtIPT7, and AtIPT8. To understand the dynamics of cytokinins in plant development, we comprehensively analyzed the expression of isopentenyltransferase genes of <jats:italic>Arabidopsis</jats:italic>. Examination of their mRNA levels and the expression patterns of the <jats:italic>beta‐glucuronidase</jats:italic> (<jats:italic>GUS</jats:italic>) gene fused to the regulatory sequence of each <jats:italic>AtIPT</jats:italic> gene revealed a specific expression pattern of each gene. The predominant expression patterns were as follows: <jats:italic>AtIPT1::GUS</jats:italic>, xylem precursor cell files in the root tip, leaf axils, ovules, and immature seeds; <jats:italic>AtIPT3::GUS</jats:italic>, phloem tissues; <jats:italic>AtIPT4::GUS</jats:italic> and <jats:italic>AtIPT8::GUS</jats:italic>, immature seeds with highest expression in the chalazal endosperm (CZE); <jats:italic>AtIPT5::GUS</jats:italic>, root primordia, columella root caps, upper part of young inflorescences, and fruit abscission zones; <jats:italic>AtIPT7::GUS</jats:italic>, endodermis of the root elongation zone, trichomes on young leaves, and some pollen tubes. <jats:italic>AtIPT1</jats:italic>, <jats:italic>AtIPT3</jats:italic>, <jats:italic>AtIPT5</jats:italic>, and <jats:italic>AtIPT7</jats:italic> were downregulated by cytokinins within 4 h. <jats:italic>AtIPT5</jats:italic> and <jats:italic>AtIPT7</jats:italic> was upregulated by auxin within 4 h in roots. <jats:italic>AtIPT3</jats:italic> was upregulated within 1 h after an application of nitrate to mineral‐starved <jats:italic>Arabidopsis</jats:italic> plants. The upregulation by nitrate did not require <jats:italic>de novo</jats:italic> protein synthesis. We also examined the expression of two genes for tRNA isopentenyltransferases, <jats:italic>AtIPT2</jats:italic> and <jats:italic>AtIPT9</jats:italic>, which can also be involved in cytokinin biosynthesis. They were expressed ubiquitously, with highest expression in proliferating tissues. These findings are discussed in relation to the role of cytokinins in plant development.</jats:p>

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