ABSTRACT
Flavonoids are secondary metabolites that fulfil a multitude of functions during the plant life cycle. In Arabidopsis proanthocyanidins (PAs) are flavonoids that specifically accumulate in the innermost integuments of the seed testa (i.e. endothelium), as well as in the chalaza and micropyle areas, and play a vital role in protecting the embryo against various biotic and abiotic stresses. PAs accumulation in the endothelium requires the activity of the MADS box transcription factor TRANSPARENT TESTA (TT) 16 (ARABIDOPSIS B-SISTER/AGAMOUS-LIKE 32) and the UDP-glycosyltransferase TT15 (UGT80B1). Interestingly tt16 and tt15 mutants display a very similar flavonoid profiles and patterns of PA accumulation. By using a combination of genetic, molecular, biochemical, and histochemical methods, we showed that both TT16 and TT15 act upstream the PA biosynthetic pathway, but through two distinct genetic routes. We also demonstrated that the activity of TT16 in regulating cell fate determination and PA accumulation in the endothelium is required in the chalaza prior to the globular stage of embryo development. Finally this study provides new insight showing that TT16 and TT15 functions extend beyond PA biosynthesis in the inner integuments of the Arabidopsis seed coat.
Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Glucosyltransferases/metabolism , MADS Domain Proteins/metabolism , Proanthocyanidins/biosynthesis , Arabidopsis/cytology , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Cell Differentiation/genetics , MADS Domain Proteins/genetics , Seeds/metabolismABSTRACT
Stomatal opening, which controls gas exchanges between plants and the atmosphere, results from an increase in turgor of the two guard cells that surround the pore of the stoma. KAT1 was the only inward K(+) channel shown to be expressed in Arabidopsis guard cells, where it was proposed to mediate a K(+) influx that enables stomatal opening. We report that another Arabidopsis K(+) channel, KAT2, is expressed in guard cells. More than KAT1, KAT2 displays functional features resembling those of native inward K(+) channels in guard cells. Coexpression in Xenopus oocytes and two-hybrid experiments indicated that KAT1 and KAT2 can form heteromultimeric channels. The data indicate that KAT2 plays a crucial role in the stomatal opening machinery.
Subject(s)
Arabidopsis Proteins , Arabidopsis/enzymology , Potassium Channels, Inwardly Rectifying , Potassium Channels/biosynthesis , Potassium Channels/genetics , Potassium Channels/metabolism , Amino Acid Sequence , Animals , Arabidopsis/genetics , Base Sequence , Cloning, Molecular , DNA, Plant/analysis , Molecular Sequence Data , Oocytes/metabolism , Plant Proteins , Potassium Channels, Voltage-Gated , RNA, Messenger/metabolism , Sequence Homology, Amino Acid , Transfection , Xenopus laevisABSTRACT
SKOR, a K+ channel identified in Arabidopsis, displays the typical hydrophobic core of the Shaker channel superfamily, a cyclic nucleotide-binding domain, and an ankyrin domain. Expression in Xenopus oocytes identified SKOR as the first member of the Shaker family in plants to be endowed with outwardly rectifying properties. SKOR expression is localized in root stelar tissues. A knockout mutant shows both lower shoot K+ content and lower xylem sap K+ concentration, indicating that SKOR is involved in K+ release into the xylem sap toward the shoots. SKOR expression is strongly inhibited by the stress phytohormone abscisic acid, supporting the hypothesis that control of K+ translocation toward the shoots is part of the plant response to water stress.
Subject(s)
Arabidopsis Proteins , Plant Proteins/isolation & purification , Potassium Channels/chemistry , Amino Acid Sequence , Animals , Arabidopsis , Cloning, Molecular , Gene Expression Regulation, Plant , Molecular Sequence Data , Plant Proteins/biosynthesis , Plant Proteins/genetics , Plant Proteins/physiology , Plant Structures/metabolism , RNA, Messenger/metabolism , RNA, Plant/metabolism , Shaker Superfamily of Potassium ChannelsABSTRACT
A ferritin cDNA, AtFer1, from seedlings of Arabidopsis thaliana has been characterized. The deduced amino acid sequence of the AtFer1 protein indicates that A. thaliana ferritin shares the same characteristics as the plant ferritin already characterized from the Leguminosae and Graminacea families: (i) it contains an additional sequence in its N-terminal part composed of two domains: a transit peptide responsible for plastid targeting and an extension peptide; (ii) amino acids that form the ferroxidase centre of H-type animal ferritin, as well as Glu residues characteristic of L-type animal ferritin, are conserved in AtFer1; (iii) the C-terminal part of the A. thaliana ferritin subunit defining the E-helix is divergent from its animal counterpart, and confirms that 4-fold-symmetry axis channels are hydrophilic in plant ferritin. Southern blot experiments indicate that AtFer1 is likely to be encoded by a unique gene in the A. thaliana genome, although a search in the NCBI dbEST database indicates that other ferritin genes, divergent from AtFer1, may exist. Iron loading of A. thaliana plantlets increased ferritin mRNA and protein abundance. In contrast to maize, the transcript abundance of a gene responding to abscisic acid (RAB18) did not increase in response to iron loading treatment, and A. thaliana ferritin mRNA abundance is not accumulated in response to a treatment with exogenous abscisic acid, at least in the culture system used in this study. In addition, iron-induced increases in ferritin mRNA abundance were the same as wild-type plants in abi1 and abi2 mutants of A. thaliana, both affected in the abscisic acid response in vegetative tissues. Increased AtFer1 transcript abundance in response to iron is inhibited by the antioxidant N-acetylcysteine. These results indicate that an oxidative pathway, independent of abscisic acid, could be responsible for the iron induction of ferritin synthesis in A. thaliana.
