RESUMO
Many plant species accumulate calcium oxalate crystals in specialised cells called crystal idioblasts. In one species of crystal-forming plants (Pistia stratiotes L.; forming raphide crystals), it has been shown that ascorbic acid is the primary precursor of oxalic acid. The question remains if this is true of other calcium oxalate crystal-forming plants. One way of answering the above question is by examining ascorbic acid as the oxalic acid precursor in diverse species with a variety of crystal types. In this study we tested ascorbic acid as the primary precursor of oxalic acid in four different species, each forming one of the four, thus far, unexamined crystal types: water hyacinth, styloid (and raphide); tomato, crystal sand; winged-bean, prismatic; water lily, astrosclereids with surface prismatic crystals. Pulse-chase feeding of 1-[14C]-ascorbic acid followed by resin embedding, microautoradiography and light microscopy were employed to examine incorporation of label into calcium oxalate crystals. For the species and crystal types studied, ascorbic acid is the primary precursor of oxalic acid and further, oxalic acid is added to crystals in patterns that correlate with the age and type of crystal involved.
RESUMO
Nep1 is an extracellular fungal protein that causes necrosis when applied to many dicotyledonous plants, including invasive weed species. Using transmission electron microscopy, it was determined that application of Nep1 (1.0 micro g mL(-)(1), 0.1% [v/v] Silwet-L77) to Arabidopsis and two invasive weed species, spotted knapweed (Centaurea maculosa) and dandelion (Taraxacum officinale), caused a reduction in the thickness of the cuticle and a breakdown of chloroplasts 1 to 4 h after treatment. Membrane breakdown was most severe in cells closest to the surface of application. Differential display was used to isolate cDNA clones from the three species showing differential expression in response to Nep1 treatment. Differential gene expression was observed for a putative serpin (CmSER-1) and a calmodulin-like (CmCAL-1) protein from spotted knapweed, and a putative protein phosphatase 2C (ToPP2C-1) and cytochrome P-450 (ToCYP-1) protein from dandelion. In addition, differential expression was observed for genes coding for a putative protein kinase (AtPK-1), a homolog (AtWI-12) of wound-induced WI12, a homolog (AtLEA-1) of late embryogenesis abundant LEA-5, a WRKY-18 DNA-binding protein (AtWRKY-18), and a phospholipase D (AtPLD-1) from Arabidopsis. Genes showing elevated mRNA levels in Nep1-treated (5 micro g mL(-)(1), 0.1% [v/v] Silwet-L77) leaves 15 min after Nep1 treatment included CmSER-1 and CmCAL-1 for spotted knapweed, ToCYP-1 and CmCAL-1 for dandelion, and AtPK-1, AtWRKY-18, AtWI-12, and AtLEA-1 for Arabidopsis. Levels of mRNA for AtPLD-1 (Arabidopsis) and ToPP2C-1 (dandelion) decreased rapidly in Silwet-L77-treated plants between 15 min and 4 h of treatment, but were maintained or decreased more slowly over time in Nep1-treated (5 micro g mL(-)(1), 0.1% [v/v] Silwet-L77) leaves. In general, increases in mRNA band intensities were in the range of two to five times, with only ToCYP-1 in dandelion exceeding an increase of 10 times. The identified genes have been shown to be involved or are related to gene families that are involved in plant stress responses, including wounding, drought, senescence, and disease resistance.
