The selection of valine-resistant mutants was carried out in leaf explant cultures of three Arabidopsis thaliana (L.) Heynh. ecotypes: C-24, RLD and Columbia. The valine concentration used for in vitro selection, lethal for seed-growing plants, has not affected callus formation and growth. However, strong inhibition of shoot regeneration ability of calli growing under selection pressure was noticed. In total, 1043 explants were cultured on valine medium and 18 shoots were regenerated with an average frequency of 1.7 shoots per 100 calli. Most R1 shoots were sterile and seeds were collected from 3 plants. The transmission of valine-resistance to the sexual progeny of these plants was scored and the increased level of valine-resistance was found in progeny of one line - 61 C. This line originated from the culture of Columbia leaf explant and displayed tetraploid chromosome number.

BACKGROUND: In the floral ABC model, B-class genes comprised of DEFICIENS (DEF)/APETALA3 (AP3) and GLOBOSA (GLO)/PISTILLATA (PI) had been proposed to involve in second and third whorl floral organ development. However, less is known about the function of B-class genes from early-diverging angiosperms. Chloranthaceae is one of the early-diverging angiosperm families. In this study, we characterized the role of the PI-like gene CsPI cloned from Chloranthus spicatus which have the simplest perianthless bisexual flowers. RESULTS: The expression profile analysis reveals high levels of CsPI mRNA in stamens in Chloranthus spicatus, with weak distribution in leaves and other floral organs. Nevertheless, CsPI rescued both stamen and petal development in Arabidopsis thaliana pi-1 mutants and caused partially conversion of sepals into petaloid organs in wild-type Arabidopsis thaliana plants. Yeast two-hybrid analysis showed that CsPI can form not only homodimers but also heterodimers with proteins encoded by Arabidopsis thaliana and Chloranthus spicatus AP3-like genes. CONCLUSIONS: These results suggested that CsPI has an ancestral function on stamen development and that CsPI has capability to specify petal development in Arabidopsis thaliana. The finding indicates that the activity of the encoded PI-like proteins is highly conserved between the early-diverging Chloranthus and Arabidopsis. Moreover, our results appear to suggest that B-function genes may not play a role in perianth development in Chloranthus spicatus.

Reactive oxygen species (ROS) have been shown to be potent signaling molecules. Today, oxidation of cysteine residues is a well-recognized posttranslational protein modification, but the signaling processes steered by such oxidations are poorly understood. To gain insight into the cysteine thiol-dependent ROS signaling in Arabidopsis thaliana , we identified the hydrogen peroxide (H ₂O ₂)-dependent sulfenome: that is, proteins with at least one cysteine thiol oxidized to a sulfenic acid. By means of a genetic construct consisting of a fusion between the C-terminal domain of the yeast (Saccharomyces cerevisiae) AP-1–like (YAP1) transcription factor and a tandem affinity purification tag, we detected ∼100 sulfenylated proteins in Arabidopsis cell suspensions exposed to H ₂O ₂ stress. The in vivo YAP1-based trapping of sulfenylated proteins was validated by a targeted in vitro analysis of DEHYDROASCORBATE REDUCTASE2 (DHAR2). In DHAR2, the active site nucleophilic cysteine is regulated through a sulfenic acid-dependent switch, leading to S-glutathionylation, a protein modification that protects the protein against oxidative damage.

The auxin‐producing bacterium Azospirillum brasilense Sp245 can promote the growth of several plant species. The model plant Arabidopsis thaliana was chosen as host plant to gain an insight into the molecular mechanisms that govern this interaction. The determination of differential gene expression in Arabidopsis roots after inoculation with either A. brasilense wild‐type or an auxin biosynthesis mutant was achieved by microarray analysis. Arabidopsis thaliana inoculation with A. brasilense wild‐type increases the number of lateral roots and root hairs, and elevates the internal auxin concentration in the plant. The A. thaliana root transcriptome undergoes extensive changes on A. brasilense inoculation, and the effects are more pronounced at later time points. The wild‐type bacterial strain induces changes in hormone‐ and defense‐related genes, as well as in plant cell wall‐related genes. The A. brasilense mutant, however, does not elicit these transcriptional changes to the same extent. There are qualitative and quantitative differences between A. thaliana responses to the wild‐type A. brasilense strain and the auxin biosynthesis mutant strain, based on both phenotypic and transcriptomic data. This illustrates the major role played by auxin in the Azospirillum–Arabidopsis interaction, and possibly also in other bacterium–plant interactions.

Arabinogalactan proteins are abundant cell-surface proteoglycans in plants and are involved in many cellular processes including somatic embryogenesis, cell-cell interactions, and cell elongation. We reported a glucuronosyltransferase encoded by Arabidopsis AtGlcAT14A, which catalyzes an addition of glucuronic acid residues to β-1,3- and β-1,6-linked galactans of arabinogalactan (Knoch et al. 2013). The knockout mutant of this gene resulted in the enhanced growth rate of hypocotyls and roots of seedlings, suggesting an involvement of AtGlcAT14A in cell elongation. AtGlcAt14A belongs to the family GT14 in the Carbohydrate Active Enzyme database (CAZy; www.cazy.org), in which a total of 11 proteins, including AtGLCAT14A, are classified from Arabidopsis thaliana. In this paper, we report the enzyme activities for the rest of the Arabidopsis GT14 isoforms, analyzed in the same way as for AtGlcAT14A. Evidently, two other Arabidopsis GT14 isoforms, At5g15050 and At2g37585, also possess the glucuronosyltransferase activity adding glucuronic acid residues to β-1,3- and β-1,6-linked galactans. Therefore, we named At5g15050 and At2g37585 as AtGlcAT14B and AtGlcAT14C, respectively.

We report that fluorescently tagged arabinogalactan glycosyltransferases target not only the Golgi apparatus but also uncharacterized smaller compartments when transiently expressed in Nicotiana benthamiana. Approximately 80% of AtGALT31A [Arabidopsis thaliana galactosyltransferase from family 31 (At1g32930)] was found in the small compartments, of which, 45 and 40% of AtGALT29A [Arabidopsis thaliana galactosyltransferase from family 29 (At1g08280)] and AtGlcAT14A [Arabidopsis thaliana glucuronosyltransferase from family 14 (At5g39990)] colocalized with AtGALT31A, respectively; in contrast, N-glycosylation enzymes rarely colocalized (3-18%), implicating a role of the small compartments in a part of arabinogalactan (O-glycan) biosynthesis rather than N-glycan processing. The dual localization of AtGALT31A was also observed for fluorescently tagged AtGALT31A stably expressed in an Arabidopsis atgalt31a mutant background. Further, site-directed mutagenesis of a phosphorylation site of AtGALT29A (Y144) increased the frequency of the protein being targeted to the AtGALT31A-localized small compartments, suggesting a role of Y144 in subcellular targeting. The AtGALT31A localized to the small compartments were colocalized with neither SYP61 (syntaxin of plants 61), a marker for trans-Golgi network (TGN), nor FM4-64-stained endosomes. However, 41% colocalized with EXO70E2 (Arabidopsis thaliana exocyst protein Exo70 homolog 2), a marker for exocyst-positive organelles, and least affected by Brefeldin A and Wortmannin. Taken together, AtGALT31A localized to small compartments that are distinct from the Golgi apparatus, the SYP61-localized TGN, FM4-64-stained endosomes and Wortmannin-vacuolated prevacuolar compartments, but may be part of an unconventional protein secretory pathway represented by EXO70E2 in plants.

The release of a reference genome for Arabidopsis thaliana in 2000 has been an enormous boon for the study of plant genetics. Less than a decade later, however, a revolution in sequencing technology had enabled rapid and inexpensive re-sequencing of whole A. thaliana genomes. Large-scale efforts to characterize natural genomic variation in A. thaliana have revealed remarkable intra-specific variation in this species, ranging from single-nucleotide differences to large structural rearrangements. The partitioning of this variation by geography and local adaptation has been described using powerful new methods and tools. Simultaneously, an ambitious research agenda has emerged to sequence 1001 A. thaliana lines from around the world, while sequencing of related species is enabling powerful evolutionary genomic analyses. In this review, I summarize recent progress in genomic analysis of natural variation in A. thaliana and its close relatives. This progress has set the stage for the emergence of Arabidopsis as a model genus for evolutionary and functional genomics.