Development of capitate glands on the leaves of annual wormwood (Artemisia annua L.) was monitored with scanning and transmission electron microscopy. Differentiation of foliar cells into gland cells began in the youngest leaf primordia. After differentiation into a 10-celled biseriate structure of two stalk cells, two basal cells, and three pairs of secretory cells, the cuticle of the six secretory cells separated from the cell walls to form a bilobed sac that eventually splits to release its contents. At every developmental stage, the cells of the the gland contained relatively little vacuolar volume. The secretory cells contained extensive endoplasmic reticulum. The plastids of each cell pair were different. At maturity, the apical cells contained proplastids or leucoplasts with only occasional thylakoids. The cell pair below the apical cell pair contained amorphous chloroplasts without starch grains. The basal cell pair contained proplastids or leucoplasts and the stalk cells contained chloroplasts. The stroma to thylakoid ratio in the secretory cell chloroplasts was high. Initially, osmiophilic product was observed most freqently associated with stacked thylakoids, plastid envelopes, and smooth endoplasmic reticulum, although it was associated with all cell membranes. Near the plasma membrane adjacent to cell walls bordering the subcuticular space, the cytoplasm was enriched in smooth endoplasmic reticulum containing osmiophilic material. The apical cell wall of the apical secretory cell pair was reticulated on the inner cytoplasmic side and contained osmiophilic staining on the cuticular side. During early senescence, osmiophilic product was commonly associated with outer mitochondrial membranes.
The root exudates produced by sorghums contain a biologically active constituent known as sorgoleone. Seven sorghum accessions were evaluated for their exudate components. Except for johnsongrass, which yielded 14.8 mg root exudate/g fresh root wt, sorghum accessions consistently yielded approximately 2 mg root exudate/g fresh root wt. Exudates contained four to six major components, with sorgoleone being the major component (> 85%). Three-dimensional structure analysis was performed to further characterize sorgoleone's mode of action. These studies indicated that sorgoleone required about half the amount of free energy (493.8 kcal/mol) compared to plastoquinone (895.3 kcal/mol) to dock into the QB-binding site of the photosystem II complex of higher plants. Light, cryo-scanning, and transmission electron microscopy were utilized in an attempt to identify the cellular location of root exudate production. From the ultrastructure analysis, it is clear that exudate is being produced in the root hairs and being deposited between the plasmalemma and cell wall. The exact manufacturing and transport mechanism of the root exudate is still unclear. Studies were also conducted on sorgoleone's soil persistence and soil activity. Soil impregnated with sorgoleone had activity against a number of plant species. Recovery rates of sorgoleone-impregnated soil ranged from 85% after 1 h to 45% after 24 h. Growth reduction of 9 14-d-old weed species was observed with foliar applications of sorgoleone.
The seed coats of S. spinosa (prickly sida, Malvaceae) become impermeable to water during seed development on the mother plant. After the seeds have dehydrated during the final maturation stages, piercing of seed coats is necessary to induce imbibition of water and germination. Onset of impermeability occurs during seed coat browning, well in advance of seed dehydration. I. Marbach and A.M. Mayer (1975, Plant Physiol. 56, 93-96) implicated polyphenol oxidase (PO; EC 1.10.3.1) as catechol oxidase in the formation of insoluble polymers during development of coat impermeability in a wild strain of pea (Pisum elatius) seeds. We found, however, that peroxidase (EC 1.11.1.7), not PO, is instrumental in the development of water-impermeable seed coats in prickly sida. We isolated coats and embryos from seeds harvested at several stages of development. Highest peroxidase activity of coat extracts correlated well with the developmental stages of maximum conversion of soluble phenolics to insoluble lignin polymers. Although seed extracts oxidized dihydroxyphenylalanine, this activity was eliminated by catalase, indicating that the oxidation of phenolics in the coat is catalyzed by peroxidase rather than PO. Histochemical localization of peroxidase was strongest in the palisade layer; both the level and time of appearance of activity was proportional to the spectrophotometric assays of seed-coat extracts. The presence of peroxidase and the absence of PO in the seed coat were also confirmed with immunocytochemistry. Our results support the view that peroxidase is involved in the polymerization of soluble phenolics to insoluble lignin polymers during development of prickly sida seed coats, causing the formation of a water-impermeable barrier prior to seed dehydration. As dehydration proceeds, the chalazal area finally becomes impermeable resulting in the hard mature seeds of prickly sida.
The effect of FMC 57020 [2-(2-chlorophenyl) methyl-4,4-dimethyl-3-isoxalidinone] on chloroplast development was examined in the cotyledons of 5-day-old, etiolated pitted morningglory (Ipomoea lacunosaL. ♯ IPOLA) seedlings grown from seeds inbibed for 24 h in water or 0.5 mM FMC 57020. In etiolated tissues, protochlorophyllide content was unaffected by FMC 57020; however, the herbicide eliminated carotenoid accumulation. There was no effect of FMC 57020 on phytoene or phytofluene content, although norflurazon [4-chloro-5-(methylamino)-2-(3-trifluoromethyl) phenyl)-3(2H)-pyridazinone] increased phytoene content in these tissues. The Shibata shift was greatly retarded in FMC 57020-treated cotyledons, suggesting that phytol levels are also reduced by the herbicide. There were no ultrastructural effects on etioplasts; however, under low white light (150 μE·m-2·s-1PAR), plastids of FMC 57020-treated seedlings did not develop into chloroplasts but rapidly developed ultrastructural symptoms of photobleaching. Starch was not mobilized in herbicide-treated plastids and sugar levels were higher in these plastids than in control plastids. Etiolated hypocotyl growth was inhibited by FMC 57020, whereas norflurazon had no effect upon it. Our results suggest that FMC 57020 blocks both diterpene and tetraterpene synthesis.
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