title: Altered cytokinin metabolism affects cytokinin, auxin, and abscisic acid contents in leaves and chloroplasts, and chloroplast ultrastructure in transgenic tobacco.
abstract: Cytokinins (CKs) are involved in the regulation of plant development including plastid differentiation and function. Partial location of CK biosynthetic pathways in plastids suggests the importance of CKs for chloroplast development. The impact of genetically modified CK metabolism on endogenous CK, indole-3-acetic acid, and abscisic acid contents in leaves and isolated intact chloroplasts of Nicotiana tabacum was determined by liquid chromatography/mass spectrometry and two-dimensional high-performance liquid chromatography, and alterations in chloroplast ultrastructure by electron microscopy. Ectopic expression of Sho, a gene encoding a Petunia hybrida isopentenyltransferase, was employed to raise CK levels. The increase in CK levels was lower in chloroplasts than in leaves. CK levels were reduced in leaves of tobacco harbouring a CK oxidase/dehydrogenase gene, AtCKX3. The total CK content also decreased in chloroplasts, but CK phosphate levels were higher than in the wild type. In a transformant overexpressing a maize beta-glucosidase gene, Zm-p60.1, naturally targeted to plastids, a decrease of CK-O-glucosides in chloroplasts was found. In leaves, the changes were not significant. CK-O-glucosides accumulated to very high levels in leaves, but not in chloroplasts, of plants overexpressing a ZOG1 gene, encoding trans-zeatin-O-glucosyltransferase from Phaseolus lunatus. Manipulation of the CK content affected levels of indole-3-acetic and abscisic acid. Chloroplasts of plants constitutively overexpressing Sho displayed ultrastructural alterations including the occasional occurrence of crystalloids and an increased number of plastoglobuli. The other transformants did not exhibit any major differences in chloroplast ultrastructure. The results suggest that plant hormone compartmentation plays an important role in hormone homeostasis and that chloroplasts are rather independent organelles with respect to regulation of CK metabolism.
authors.affiliation: Institute of Experimental Botany AS CR, Rozvojová 135, 165 02 Prague 6-Lysolaje, Czech Republic. polanska@ueb.cas.cz
authors.forename: Lenka
authors.initials: L
authors.lastname: Polanská
authors.forename: Anna
authors.initials: A
authors.lastname: Vicánková
authors.forename: Marie
authors.initials: M
authors.lastname: Nováková
authors.forename: Jirí
authors.initials: J
authors.lastname: Malbeck
authors.forename: Petre I
authors.initials: PI
authors.lastname: Dobrev
authors.forename: Bretislav
authors.initials: B
authors.lastname: Brzobohaty
authors.forename: Radomíra
authors.initials: R
authors.lastname: Vanková
authors.forename: Ivana
authors.initials: I
authors.lastname: Machácková
mesh_terms: D000040:Abscisic Acid / Q000378:metabolism*; D019883:Alkyl and Aryl Transferases / Q000235:genetics / Q000378:metabolism; D002736:Chloroplasts / Q000378:metabolism / Q000648:ultrastructure; D003583:Cytokinins / Q000378:metabolism*; D005964:Glucosyltransferases / Q000235:genetics; D007210:Indoleacetic Acids / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D032306:Petunia / Q000235:genetics; D027805:Phaseolus / Q000235:genetics; D018515:Plant Leaves / Q000378:metabolism / Q000648:ultrastructure; D010940:Plant Proteins / Q000235:genetics; D030821:Plants, Genetically Modified / Q000378:metabolism* / Q000648:ultrastructure; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D003313:Zea mays / Q000235:genetics; D001617:beta-Glucosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D003583:Cytokinins; D007210:Indoleacetic Acids; D010940:Plant Proteins; C030737:indoleacetic acid; D000040:Abscisic Acid; D010088:Oxidoreductases; C019984:cytokinin oxidase; D005964:Glucosyltransferases; C117387:UDPglucose zeatin O-glucosyltransferase, plant; D019883:Alkyl and Aryl Transferases; C024274:adenylate isopentenyltransferase; D001617:beta-Glucosidase