The interactions of paraquat (methylviologen) and diphenylether herbicides with the Mehler reaction as investigated. Sera from two different rabbits (RS1 & RS2) were examined for their patterns of inhibition of the photosynthetic electron transport (PET) system. Serum from RS2 was greatly hemolyzed. Fifty ul of RS1 serum were required for 100% inhibition of a H₂O → methylviologen(MV)/O₂ reaction, whereas only 10 µl of a 1:10 dilution of RS2 were needed for 100% inhibition. The γ-globulin fraction from purified rabbit serum (RS1) did not inhibit PET, indicating that the antibody fraction of the rabbit serum does not contain the inhibitor. It appears that the inhibitor is from the hemolyzed red blood cells. Rabbit sera, added to chloroplast preparations prior illumination, caused no inhibition of a H₂O → MV/O₂ reaction while addition of rabbit sera during illumination inhibited the H₂O → MV/O₂ reaction within 1-3 s. Various Hill reactions were used to determine the site of inhibition. Rabbit sera inhibited photosystem I (PSI) Hill reactions, but did not inhibit a photosystem II (PSI II) Hill reaction indicating that inhibition is on the reducing side of PSI. It would be expected that a H₂O → Ferredoxin (Fd)/NADP Hill reaction should also be blocked. Surprisingly, rabbit sera did not inhibit this reaction. These results were interpreted as supportive evidence for parallel (branched) electron transport on the reducing side of PSI. Six pyridyl derivatives {benzylviologen, 2-anilinopyridine, 1,2-bis(4-pyridyl)ethane, 1,2-bis(4-pyridyl)ethylene, 2-benzoylpyridine, and 2-benzylaminopyridine} and five heme-iron derivatives {hemoglobin, hemin, hematin, ferritin, and ferrocene} were screened for their potential to counteract paraquat toxicity on pea (Pisum sativum L. cv. Little Marvel) isolated chloroplasts. H₂O → MV/O₂ and H₂O → Fd/NADP+ were the two Hill reactions assayed with these compounds. Antagonists of paraquat toxicity should inhibit the first Hill reaction but not the latter. None of the pyridyl derivatives examined inhibited the reaction H₂O → MV/O₂. Ferritin and ferrocene were also ineffective as inhibitors of this reaction. Hemoglobin inhibited the reaction H₂O → MV/O₂ without inhibiting the reaction H₂O → Fd/NADP+, providing protection to pea chloroplasts against paraquat. Hemin and hematin inhibited both Hill reactions examined. Hemin and hematin also inhibited H₂O → diaminodurene (ox) and durohydroquinone → MV/O₂ Hill reactions but not the dichlorophenylindolphenol(red) → MV/O₂ and diaminodurene(red) → MV/O₂ Hill reactions. These results indicate that hemin and hematin are inhibiting photosynthetic electron transport in the plastoquinone pool region. Potential involvement of hydroxyl and alkoxyl radicals in the peroxidative action of the p-nitro diphenyl ether herbicides acifluorfen was evaluated under laboratory conditions. Methional was added to illuminated pea thylakoids and its oxidation to ethylene was used as an indicator of hydroxyl and alkoxyl radical synthesis. Oxyfluorfenstimulation of the rate of methional oxidation was dependent on light, photosynthetic electron transport and hydrogen peroxide since it was not observed under dark conditions or in the presence of DCMU and catalase. Addition of FeEDTA, a catalyst of the Fenton reaction, stimulated the oxyfluorfen-induced enhancement of methional oxidation six-fold suggesting that hydroxyl radicals are synthesized through a Fenton reaction. Acifluorfen, nitrofen and nitrofluorfen inhibited the rate of methional oxidation whereas, acifluorfen-methyl had no effect on the rate of methional oxidation even at high concentrations (1 mM). Nitrofluorfen at 1 mM was the only p-nitro diphenyl ether herbicide tested which inhibited photosynthetic electron transport of pea thylakoids. In experiments with pea leaf discs, acifluorfen at low concentrations stimulated the rate of methional oxidation, while acifluorfen-methyl, nitrofen and nitrofluorfen had no effect. These data indicate that hydroxyl and alkoxyl radicals could be involved in the mechanism of cellular damage caused by oxyfluorfen, but they are not important for the activity of the diphenyl ether herbicides acifluorfen, acifluorfen-methyl, nitrofen, and nitrofluorfen. Diethyldithiocarbamate (DEDTC) does not accept electrons from the photosynthetic electron transport (PET), but can donate electrons to a photosystem I (PSI) Mehler reaction in the presence of the following PET inhibitors: diuron, dibromothymoquinone, and bathophenanthroline. It cannot photoreduce PSI in the presence of cyanide, a PET inhibitor. These data indicate that the site of electron donation is after the plastoquinone pool. Ascorbate is not required for the ability of DEDTC to donate electrons to PSI. There is no photoreductant activity by DEDTC in a ferredoxin/NADP Hill reaction. Superoxide dismutase inhibits DEDTC/diuron or bathophenanthroline → MV/O₂ Mehler reaction. Catalase does not restore the consumed O₂ from a DEDTC/diuron → MV/O₂ Mehler reaction, indicating O₂- has not been dissmutating into H₂O₂. These results indicate that superoxide is required for DEDTC ability to donate electrons, therefore DEDTC is limited only to Mehler-type reactions. / Ph. D.

A portable infra red gas analyzer was used to investigate the interactive effects of light, temperature, and CO,jO, ratios under controlled environmental conditions in an attempt to model gas exchange characteristics of Coix Iachryma-jobi L. Plotting light response curves as a function of temperature (20, 25 30 and 35°C) revealed no sign of light saturation even at a photosynthetic photon flux density (PPFD) close to 2000 !Lmol m-' sol. High net assimilation rates (A) of approximately 24 !Lmol CO, m"s'! were realized at 30-35°C. Assimilation (A) versus internal CO, partial pressure (C,) curves showed a steep rise with increase in C, but saturated at approximately 150 (JLII-!) and all the results, either in the absence or presence of 0" showed a similar response under all temperature regimes. C. Iachryma-jobi exhibited low CO, compensation points cr ) between 0 and 10 JLlI-! under similar experimental temperatures and either at 0 or 21%0,. The slopes of double reciprocal plots of llA versus llCi, were nearly identical and crossed the yintercept at almost identical points under all 0, concentrations. These data indicate first; that there was no apparent 0, inhibition and second; indicated that the apparent inhIbitor constant (K,) for 0, at the site of carboxylation did not change with increase in [OJ from 0 to 21% oxygen. These observations were further confirmed by results obtained from the analysis of apparent carboxylation efficiency (CE, as defined as the slope of response of A to increasing CO,), as no inhibition of A with increase of [OJ occurred. These characteristics were consistent with typical features of C,photosynthesis. The absence of 0, inhibition and low r values indicated that an efficient CO, concentrating mechanism which eliminates photorespiration exists in C. Iachryma-jobi. At the light microscope level, leaf anatomy exhibited typical C, structure viz. bundle sheath with large chloroplasts and this sheath is further surrounded by a radiate Kranz mesophyU cells. Furthermore the anatomical features suggested that C. wchryma-jobi was an NADP-ME species. Stomatal conductance (g,) to assimilation (g,/A) indicated an increase in A with decrease in g" an essential feature of improving water use efficiency (WUE), but one which drastically reduces CO, diffusion rate. The physical lintitation (stomatal lintitation, t) to CO, diffusion under various [0,] and temperatures, but constant PPFD, did not exhibit statistically significant change in t values at either 0 or 21% a, within each temperature regime, however there was a marked decrease in t as the plant approached its optimum photosynthetic temperature.