Using degenerate primers designed by deduced amino acid sequences of known aldehyde oxidases (AO) from maize and bovine, two independent cDNA fragments were amplified by reverse transcription-polymerase chain reaction (PCR). The two corresponding full-length cDNAs (atAO-1 and atAO-2; 4,484 and 4,228 bp long, respectively) were cloned by screening the Arabidopsis cDNA library followed by rapid amplification of cDNA end-PCR. These cDNAs are highly homologous at both the nucleotide and amino acid sequence levels, and the deduced amino acid sequences showed high similarity with those of maize and tomato AOs. They contain consensus sequences for two iron-sulfur centers and a molybdenum cofactor (MoCo)-binding domain. In addition, another cDNA having a sequence similar to that of the cDNAs was screened (atAO-3; 3,049 bp), and a putative AO gene (AC002376) was reported on chromosome 1, which (atAO-4) was distinct from, but very similar to, the above three AOs. atAO-1, 2, 3, and 4 were physically mapped on chromosomes 5, 3, 2 and 1, respectively. These data indicate that there is an AO multigene family in Arabidopsis. atAO-1 protein was shown to be highly similar to one of the maize AOs in respect to a region thought to be involved in determination of substrate specificity, suggesting that they might encode a similar type of AO, which could efficiently oxidize indole-3-acetaldehyde to indole-3-acetic acid (IAA). atAO-1 and atAO-2 genes were expressed at higher levels in lower hypocotyls and roots of the wild-type seedlings, while atAO-3 was slightly higher in cotyledons and upper hypocotyls. The expression of atAO-1 was more abundant in the seedlings of an IAA overproducing mutant (superroot1; sur1) than in those of wild type. atAO-2 and atAO-3 transcripts were rather evenly distributed in these seedlings. A possible involvement of atAO genes in phytohormone biosynthesis in Arabidopsis is discussed.

Abscisic acid (ABA) is involved in seed development and plant adaptation to environmental stresses. ABA is synthesized from cleaved xanthophylls and zeaxanthin epoxidase (ZEP) is the enzyme responsible for the conversion of zeaxanthin to violaxanthin. In this study, we have characterized the ABA1 gene (AtZEP) of Arabidopsis thaliana L. and show that this complements the aba1 mutant, defective in zeaxanthin epoxidation. The molecular basis for two aba1 mutant alleles has been determined and the reduction in their AtZEP transcript levels correlates with the molecular defect identified. As AtZEP mRNA abundance was not affected in two other ABA-deficient mutants (aba2 and aba3) and in two ABA-insensitive mutants (abi1 and abi2), no feedback regulation of ABA biosynthesis seems to occur at the level of ZEP transcription. Steady state transcript levels increased in roots during rapid water stress as well as progressive drought stress, providing evidence that zeaxanthin epoxidation contributed to the regulation of ABA biosynthesis in roots and consequently to the plant adaptive response to hydric stress. In seeds in situ hybridization analysis detectedAtZEP mRNA in the embryo cells from the globular stage to desiccation phase. In contrast, expression of AtZEP in maternal tissues was specific to the maturation phase. These results are discussed in relation to the role of ABA both in response to drought stress and in seed development.

Besides these differences in cell pigmentation, a rapid but transient accumulation of cyanophycin granule polypeptide occurred in ammoniumgrown cells, while these macromolecules were not detected in cells grown with nitrate. In contrast, glycogen reserves displayed a dynamic pattern of accumulation and disappearance during cell growth which varied only slightly with the nitrogen source. The observed changes in cell pigmentation are reminiscent of the phenomenon of complementary chromatic adaptation, in which green and red wavelengths promote the syntheses of phycoerythrin and phycocyanin-2, respectively. As in complementary chromatic adaptation, the regulation of synthesis of phycoerythrin and phycocyanin-2 by the nitrogen source occurred mainly at the mRNA level. Moreover, the transcriptional start sites for the expression of the cpeBA and the cpc2 operons, which respectively encode the two subunits of phycoerythrin and phycocyanin-2, were the same in cells grown in nitrate or ammonium, and identical to those in green-and red-light-grown cells. The results of this study suggest that acclimation to the spectral light quality and to the nitrogen source share some common regulatory elements.

No abstract

In the absence of a tight control of copper entrance into cells, bacteria have evolved different systems to control copper concentration within the cytoplasm and the periplasm. Central to these systems, the Cu+ ATPase CopA plays a major role in copper tolerance and translocates copper from the cytoplasm to the periplasm. The fate of copper in the periplasm varies among species. Copper can be sequestered, oxidized, or released outside the cells. Here we describe the identification of CopI, a periplasmic protein present in many proteobacteria, and show its requirement for copper tolerance in Rubrivivax gelatinosus. The ΔcopI mutant is more susceptible to copper than the Cu+ ATPase copA mutant. CopI is induced by copper, localized in the periplasm and could bind copper. Interestingly, copper affects cytochrome c membrane complexes (cbb3 oxidase and photosystem) in both ΔcopI and copA-null mutants, but the causes are different. In the copA mutant, heme and chlorophyll synthesis are affected, whereas in ΔcopI mutant, the decrease is a consequence of impaired cytochrome c assembly. This impact on c-type cytochromes would contribute also to the copper toxicity in the periplasm of the wild-type cells when they are exposed to high copper concentrations.