Structure and expression of 12-oxophytodienoate reductase (OPR) subgroup I gene in pea and oxidoreductase activity of their recombinant proteins

Recently, we observed that expression of a pea gene (S64) encoding an oxophytodienoic acid reductase (OPR) was induced by a suppressor of pea defense responses, secreted by the pea pathogen Mycosphaerella pinodes. Because it is known that OPRs are usually encoded by families of homologous genes, we screened for genomic and cDNA clones encoding members of this putative OPR family in pea. We isolated five members of the OPR gene family from a pea genomic DNA library, and amplified six cDNA clones, including S64, by RT-PCR (reverse transcriptase-PCR). Sequencing analysis revealed that S64 corresponds to PsOPR2, and the amino acid sequences of the predicted products of the six OPR-like genes shared more than 80% identity with each other. Based on their sequence similarity, all these OPR-like genes code for OPRs of subgroup I, i.e., enzymes which are not required for jasmonic acid biosynthesis. However, the genes varied in their exon/intron organization and in their promoter sequences. To investigate the expression of each individual OPR-like gene, RT-PCR was performed using gene-specific primers. The results indicated that the OPR-like gene most strongly induced by the inoculation of pea plants with a compatible pathogen and by treatment with the suppressor from M. pinodes was PsOPR2. Furthermore, the ability of the six recombinant OPR-like proteins to reduce a model substrate, 2-cyclohexen-1-one (2-CyHE), was investigated. The results indicated that PsOPR1, 4 and 6 display robust activity, and PsOPR2 has a most remarkable ability to reduce 2-CyHE, whereas PsOPR3 has little and PsOPR5 does not reduce this compound. Thus, the six OPR-like proteins can be classified into four types. Interestingly, the gene structures, expression profiles, and enzymatic activities used to classify each member of the pea OPR-like gene family are clearly correlated, indicating that each member of this OPR-like family has a distinct function.</p

Modulation of defense signal transduction by flagellin-induced WRKY41 transcription factor in Arabidopsis thaliana

Flagellin, a component of the flagellar filament of Pseudomonas syringae pv. tabaci 6605 (Pta), induces hypersensitive reaction in its non-host Arabidopsis thaliana. We identified the WRKY41 gene, which belongs to a multigene family encoding WRKY plant-specific transcription factors, as one of the flagellin-inducible genes in A. thaliana. Expression of WRKY41 is induced by inoculation with the incompatible pathogen P. syringae pv. tomato DC3000 (Pto) possessing AvrRpt2 and the non-host pathogens Pta within 6-h after inoculation, but not by inoculation with the compatible Pto. Expression of WRKY41 was also induced by inoculation of A. thaliana with an hrp-type three secretion system (T3SS)-defective mutant of Pto, indicating that effectors produced by T3SS in the Pto wild-type suppress the activation of WRKY41. Arabidopsis overexpressing WRKY41 showed enhanced resistance to the Pto wild-type but increased susceptibility to Erwinia carotovora EC1. WRKY41-overexpressing Arabidopsis constitutively expresses the PR5 gene, but suppresses the methyl jasmonate-induced PDF1.2 gene expression. These results demonstrate that WRKY41 may be a key regulator in the cross talk of salicylic acid and jasmonic acid pathways.</p

Geochemistry of surface sediments in tsunami-affected Sri Lankan lagoons regarding environmental implications

The December 26, 2004 Indian Ocean tsunami was one of the largest in human history, devastating the coastal wetlands of surrounding countries. This study present the chemical analyses of tsunamigenic and pre-tsunami sediments from Hikkaduwa and Hambantota lagoons in southern Sri Lanka, to assess their geochemical composition, their source, and subsequent environmental impacts. Principal component analysis of the tsunami sediments shows that 42% of the total variance is accounted for calcium oxide and Sr. That is, the tsunami deposits are rich in biogenic phases derived from shallow marine sediments. High organic matter contents of the tsunami sediments of up to 80 wt% also support this interpretation. The association of chlorine (<9.4 wt%), brome (<170 mg/kg), arsenic (<17 mg/kg), iron (III) oxide (<12.9 wt%) and sulfur (<7.6 wt%) accounts for 33% of the variance, reflecting higher salinity. This further suggests that the sediments were mainly derived from a marine environment, rather than from non-marine sands and/or soils. Immobile element contents and relations (thorium, scandium and zirconium) suggest that the tsunami sediment source was mostly felsic in composition, with some mafic component, and mixed with predominantly shallow marine shelf or slope sediments. Additional compositional variations in the tsunami sediments in both lagoons may be associated with variations of wave strength along the coast and by the morphology of the continental shelf. Lower elemental abundances in Hambantota lagoon sediments compared to Hikkaduwa equivalents may thus reflect a greater non-marine component in the former, and greater shelf sediment component in the latter