Efficient photosynthesis requires a balance of ATP and NADPH production/consumption in chloroplasts, and the exportation of reducing equivalents from chloroplasts is important for balancing stromal ATP/NADPH ratio. Here, we showed that the overexpression of purple acid phosphatase 2 on the outer membranes of chloroplasts and mitochondria can streamline the production and consumption of reducing equivalents in these two organelles, respectively. A higher capacity of consumption of reducing equivalents in mitochondria can indirectly help chloroplasts to balance the ATP/NADPH ratio in stroma and recycle NADP+, the electron acceptors of the linear electron flow (LEF). A higher rate of ATP and NADPH production from the LEF, a higher capacity of carbon fixation by the Calvin–Benson–Bassham (CBB) cycle and a greater consumption of NADH in mitochondria enhance photosynthesis in the chloroplasts, ATP production in the mitochondria and sucrose synthesis in the cytosol and eventually boost plant growth and seed yields in the overexpression lines.

A study of the C4 plant itchgrass (Rottboellia exaltata L.f.) grown under 100%, 60%, 25% and 2% sunlight revealed differences in the anatomy and cytology of the foliar mesophyll and vascular bundles associated with shading. In the bundle sheath, shading caused a reduction in thickness of the cell walls, shrinkage of plastids, rearrangement of plastid thylakoids, a reduction in starch deposits and vacuolization of the cytoplasm. In general, plastids and mitochondria retained membrane integrity but underwent stromal deterioration. Shading effects on cytoplasm were similar for mesophyll and bundle sheath cells. Mesophyll chloroplasts lost starch grains and the peripheral reticulum tended to decrease with greater shading. Grana were well developed at all irradiances, although the chloroplasts themselves decreased in size with shading. Movement of mesophyll chloroplasts away from bundle sheath-mesophyll borders was pronounced at 60% sunlight and was progressively greater at the two lower irradiances. These observations suggest that the structural relationships thought to be necessary for the intercellular transfer of C4 acids and the functioning of the C4 photosynthetic pathway were disrupted by shading.

A procedure is described for the rapid analysis of leaf samples to determine triazine resistance. The technique uses a commercially available fluorometer and is based upon the fact that photosynthesis-inhibiting herbicides increase chlorophyll fluorescence (because of dissipation of absorbed radiant energy in the absence of useful photochemistry). Resistant and susceptible biotypes of six weed species were assayed. Fluorescence of susceptible leaf sections increased dramatically over a 1- to 3-h exposure to atrazine [2-chloro-4-(ethylamino)-6-(isopropylamino)-s-triazine], but resistant leaf sections showed no fluorescence increase. Substantial fluorescence increases of both resistant and susceptible leaf sections were induced by diuron [3-(3,4-dichlorophenyl)-1,1-dimethylurea]. In the absence of herbicides, fluorescence was higher in resistant than in susceptible leaf sections, suggesting that the resistant biotypes are less efficient photosynthetically. Fluorescence analysis was used to characterize atrazine inhibition of photosynthesis in leaf sections of three crop species. Differences in atrazine-induced fluorescence between crop lines were relatively small and were not well correlated with atrazine tolerance. The fluorometer is a convenient device for monitoring photosynthesis inhibition, however, and could be useful in detecting plants having resistance to photosynthesis-inhibiting herbicides.

Suspected triazine resistant Pennsylvania smartweed plants were discovered near Holy Cross, Dubuque County, IA. Subsequent variable chlorophyll a fluorescence assays confirmed these selections to be triazine resistant. This is the first reported evidence of s-triazine resistance in Pennsylvania smartweed.

The effects of oxyfluorfen [2-chloro-1-(3-ethoxy-4-nitrophenoxy)-4-(trifluoromethyl)benzene] were studied on electron transport and phosphorylation in isolated spinach (Spinacia oleracea L.) chloroplasts and on the response of green bean (Phaseolus vulgaris L. ‘Spartan Arrow’) to applications of oxyfluorfen alone or in combination with other herbicides. Coupled non-cyclic electron transfer and phosphorylation through photosystems I and II (H2 O å methyl viologen) were inhibited about 30% and 55%, respectively, by 10−4 M oxyfluorfen. Photosystem II-linked phosphorylation with dimethylbenzoquinone (2,5-dimethyl-p-benzoquinone) as the electron acceptor (H2 O å DMQ) was completely inhibited by 10−4 M oxyfluorfen. Photosystem II electron transport with dimethylbenzoquinone as the electron acceptor was inhibited 60% by 10−4 M oxyfluorfen, whereas photosystem I electron transport with 2,6-dichlorophenol indophenol as electron donor (DCIPH2 å MV) was not susceptible to oxyfluorfen inhibition. Photosystem I-linked phosphorylation and the accompanying electron transport supported by durohydroquinone electron donation (DQH2 å MV) were inhibited about 50% by 10−4 M oxyfluorfen, whereas cyclic phosphorylation was not inhibited at that concentration. Increased conductivity of a solution that contained leaf discs taken from green beans treated with various combinations of foliar-applied herbicides was a measure of membrane damage caused by the herbicides, and revealed that oxyfluorfen has a different site of action than do photosynthesis inhibitor and bipyridilium herbicides. Oxyfluorfen plus dinoseb (2-sec-butyl-4,6-dinitrophenol) injury to green beans was additive, but the two herbicides did not have the same site of action. Oxyfluorfen did not appear to inhibit electron transport in chloroplasts at herbicidal rates, nor was it dependent on electron transport for activation.

We undertook a study of Porphyra acanthophora var. brasiliensis to determine its responses under ambient conditions, photosynthetically active radiation (PAR), and PAR+UVBR (ultraviolet radiation-B) treatment, focusing on changes in ultrastructure, and cytochemistry. Accordingly, control ambient samples were collected in the field, and two different treatments were performed in the laboratory. Plants were exposed to PAR at 60 μmol photons m−2 s−1 and PAR + UVBR at 0.35 W m−2 for 3 h per day during 21 days of in vitro cultivation. Confocal laser scanning microscopy analysis of the vegetative cells showed single stellate chloroplast in ambient and PAR samples, but in PAR+UVBR-exposed plants, the chloroplast showed alterations in the number and form of arms. Under PAR+UVBR treatment, the thylakoids of the chloroplasts were disrupted, and an increase in the number of plastoglobuli was observed, in addition to mitochondria, which appeared with irregular, disrupted morphology compared to ambient and PAR samples. After UVBR exposure, the formation of carpospores was also observed. Plants under ambient conditions, as well as those treated with PAR and PAR+UVBR, all showed different concentrations of enzymatic response, including glutathione peroxidase and reductase activity. In summary, the present study demonstrates that P. acanthophora var. brasiliensis shows the activation of distinct mechanisms against natural radiation, PAR and PAR+UVBR.

Ruminant products are criticised for their SFA content relative to PUFA, although n-6:n-3 PUFA is desirable for human health ( < 4). Rumen protozoa are rich in unsaturated fatty acids due to engulfment of PUFA-rich chloroplasts. Increasing the chloroplast content of rumen protozoa offers a potentially novel approach to enhance PUFA flow to the duodenum and subsequent incorporation into meat and milk. We evaluated protozoal contribution to duodenal n-3 PUFA flow due to intracellular chloroplast content. A total of six Holstein × Friesian steers were fed, in a two-period changeover design, either straw:concentrate (S:C, 60:40; DM basis; S:C, low chloroplast) or fresh perennial ryegrass (PRG; high chloroplast). Following 12 d adaptation to diet, ruminal protozoal and whole duodenal samples were obtained. N and fatty acid content of whole duodenum and rumen protozoal samples were assessed and protozoal 18S rDNA quantitative PCR performed, enabling calculation of protozoal N flow. The ratio of individual fatty acids:N in rumen protozoal samples was calculated to obtain protozoal fatty acid flows. Based on total fatty acid flow, contribution (%) of protozoa to individual fatty acid flows was calculated. Protozoal fatty acid data and microscopical observations revealed that protozoa were enriched with 18 : 3n-3 following PRG feeding, compared with the S:C diet, due to increased intracellular chloroplast content. However, duodenal protozoal 18S rDNA concentration post PRG feeding was low, indicating rumen retention of the protozoa. Nutrition influences the 18 : 3n-3 content of protozoa; the challenge is to increase protozoal flow to the small intestine, while maintaining sustainable rumen densities.

Twenty-five years ago, a basic tenet of biochemistry held that enzymes are made solely of protein. Ribosomes, which are large ribonucleoprotein enzymes, were conveniently overlooked (but see Woese, 1967; Crick, 1968; Orgel, 1968). The then current orthodoxy was first challenged by the discovery that Escherichia coli RNase P had an essential RNA subunit (Stark et al., 1978) and completely demolished when the catalytic properties of RNA were described (Kruger et al., 1982; Guerrier-Takada et al., 1983; Buzayan et al., 1986; Hutchins et al., 1986). With respect to RNase P, in particular, a new orthodoxy arose: RNase P from any source would have an essential RNA subunit although not necessarily a catalytic one. This adolescent orthodoxy is now being challenged by reports of putative RNase P activities isolated from certain organelles that lack an RNA subunit (Manam & van Tuyle, 1987; Rossmanith & Karwan, 1998; Wang et al., 1998). The following questions must be asked of the aforementioned data: (1) Are the substrate specificities the same as in the canonical reaction? (2) Are the chemical mechanisms of the newly identified “RNase P” reactions identical to the canonical reaction? (3) Have completely purified (i.e., cloned, expressed, and/or purified) activities been studied? (4) Have the phenotypes of these “new” enzymes been verified by genetic means? (5) In sum, are we dealing with RNase P or some other activities that share some characteristics with RNase P? Should these be called RNase P, in any case?

Effects of ozone on spring wheat (Triticum aestivum L. cv. Satu) were studied in an open-top chamber experiment during two growing seasons (1992–1993) at Jokioinen in south-west Finland. The wheat was exposed to filtered air (CF), non-filtered air (NF), non-filtered air+35 nl l−1 ozone for 8 h d−1 (NF+) and ambient air (AA). Each treatment was replicated five times. Two wk after anthesis, after 4 wk of ozone treatment (NF+, 45 nl l−1 1000–1800 hours, seasonal mean) the net CO2 uptake of wheat flag leaves was decreased by c. 40% relative to CF and NF treatments, both initial and total activity of Rubisco and the quantity of protein-bound SH groups were decreased significantly. Added ozone also significantly accelerated flag leaf senescence recorded as a decrease in chloroplast size. The effect was significant 2 wk after anthesis, and senescence was complete after 4 wk. In the CF and NF treatments senescence was complete 5 wk after anthesis. The significant effect of ozone on the chloroplasts and net CO2 uptake 2 wk after anthesis did not affect the grain filling rate. However, since the grain filling period was shorter for ozone fumigated plants, kernels were smaller. The decrease in 1000-grain weight explained most of the yield reduction in the plants under NF+ treatment. The results indicate that wheat plants are well buffered against substantial decrease in source activity, and that shortened flag leaf duration is the major factor causing ozone-induced yield loss.