mitochondria electron transport inhibitors or METIs ( Ware and Whitacre, 2004 ; Yu, 2008 ). The mitochondrion (plural: mitochondria) is a membrane-bounded organelle that is associated with intracellular respiration. The mitochondrion is a major site of
Mitochondria isolated from the pericarp tissue of green bell pepper (Capsicum annuum L.) fruit and purified on a Percoll gradient produced superoxide in buffers aerated with oxygen. ADP and uncouplers of the electron transport chain reduced superoxide production. Disulfiram, an inhibitor of the alternative oxidase, enhanced superoxide production. Inhibitors of the ubiquinone-cytochrome bc1 complex had little effect on superoxide production by mitochondria which were insensitive to cyanide. Less superoxide was produced when DTT was used to reduce the sulfhydryl groups of the alternative oxidase protein and the enzyme was activated with pyruvate than when the sulfhydryl groups were oxidized with diamide. A role for the alternative oxidase in limiting the level of reactive oxygen species produced in stressed and senescing plant tissues is suggested.
An emerging theory contends that chilling injury is due to oxidative damage resulting from the metabolic generation of active oxygen species. Mitochondria were isolated from chilling-sensitive (CS) and from conditioned chilling-resistant (CR) bell pepper fruit and their ability to generate superoxide determined by measuring the formation of adrenochrome from epinephrine. Mitochondria from CS fruit were sensitive to cyanide and produced superoxide when supplied with NADH, succinate, or malate-pyruvate. Mitochondria from CR fruit were insensitive to cyanide and sensitive to SHAM and produced little superoxide when supplied with respiratory substrates. ATP enhanced the production of superoxide and ADP reduced the production. Results suggest that the mitochondria are a major source of superoxide in CS plant tissue and the presence of the alternative pathway reduces the production of superoxide.
Reactive O2 species produced when electron transport is disrupted have been implicated in several environmental stress-induced disorders. Superoxide (
A strong association is implicit between mitochondrial function and the energy demands of cells responding to stress. Yet, the dynamics of this organelle-cellular dependency have been difficult to resolve. This study examines a new diagnostic parameter namely, mitochondrial maintenance and self-restoration as exhibited by the course of respiratory functions (states 3 and 4 respiratory rates, respiratory control) of mitochoudria extracted during and after exposure of intact `Hass' avocado (Persea americana) fruit to different stress atmospheres: anoxia (100% N2) or high (25% and 75%) CO2 for varying durations. Comparisons are made with direct exposure of the mitochondria themselves to similar atmospheres. In general, exposure of the fruit to CO2 rich atmospheres enhanced the capacity of their mitochondria to restore energy-linked functions whereas anoxia caused irreparable damage. The physiological (climacteric) state of the fruit also affected the stress capacity of the mitochondria contained therein, anaerobiosis being more harmful to mitochondria in riper fruit. In contrast to their effects in vivo, in vitro anoxia appeared to sustain mitochondrial energy-linked functions, whereas high CO2 was clearly harmful. These and other observations are discussed in the context of mitochondrial self-restoration or homeostasis and its relevance to postharvest stress-atmosphere storage for purposes such as pathogen suppression or insect control.
Chilling temperatures (≤12°C) can cause substantial economic damage to cucumber (Cucumis sativus L.) plants. Previous studies suggest chilling tolerance trait is controlled by nuclear gene(s). To investigate inheritance of chilling injury in cucumber, cucumber lines [susceptible GY14 (P1), tolerant `Chipper' (P2), and tolerant `Little John' (P3)], and their exact reciprocal F1 and F2 cross-progeny were evaluated to determine the inheritance of chilling injury at the first true-leaf stage when challenged at 4 °C for 5.5 hours. The mean chilling ratings [1(trace) to 9(dead)] of progeny comparisons were F1(P1 × P2) = 6.2 vs. F1(P2 × P1) = 1.6; F2(P1 × P2) = 6.4 vs. F2(P2 × P1) = 2.7; F1(P1 × P3) = 5.4 vs. F1(P3 × P1) = 1.7; and F2(P1 × P3) = 5.8 vs. F2(P3 × P1) = 2.2. These data suggest that chilling tolerance was maternally inherited as is the chloroplast genome in cucumber. Parents, reciprocal F1, and F2 progeny were evaluated for variation using random amplified polymorphism DNA (RAPD). Although no maternally inherited RAPD markers were detected, polymorphic and paternally inherited RAPD bands AD21249, AV8916, and AV8969 amplified by AD2 and AV8 primers were cloned and sequenced. A BLAST search of these sequences suggested that their origin is likely cucumber mitochondrial DNA. These results indicate that the mitochondria genome is not associated with the chilling tolerant trait because this genome is paternally inherited in progeny derived from this reciprocal mating. Therefore, the results of maternally inherited chilling tolerant trait and paternally transmitted mitochondria genome support that the chilling tolerant trait as identified is likely associated with the chloroplast genome which is maternally transmitted in cucumber.
Abstract
Cyanide-resistant respiration in carnation (Dianthus caryophyllus L.) petal tissue increased daily until 3 days after flowers were harvested. Mitochondria from petals exhibited only cyanide-resistant respiration when malate and pyruvate were provided as substrate. Although intact petals from freshly harvested flowers exhibited little cyanide-resistant respiration, mitochondria isolated from them were highly cyanide-resistant in their respiration if malate and pyruvate were provided as substrate. Pyruvate, in mitochondria, was capable of releasing mitochondria inhibited by cyanide in the presence of malate. Pentose-phosphate pathway activity was significant throughout the postharvest period, but was maximum 5 days after flowers were harvested. The gas exchange quotient (GEQ) was 1 for the first 3 days after harvest, declining on days 4 and 5 and dramatically increasing on day 6 as a result of a climacteric increase in CO2 evolution accompanying flower wilting.
intergenic regions in the mitochondria of carrot ( Daucus carota L.) ( Bach, 2000 ) were used. Primers primarily targeted mitochondrial genes, but several chloroplast gene primers were included because they occur in the carrot mitochondrial genome ( Bach
The mechanism by which diphenylamine (DPA) controls superficial scald in apples and reduces chilling injury in green bell peppers [Capsicum annuum L. var. annuum (Grossum Group)] has been assumed to be related to its antioxidant activity. In the present study, DPA inhibited the respiratory activity of green bell pepper fruit as well as oxygen uptake by the mitochondria isolated from them. When the alternative oxidase was inhibited with n-propyl gallate or disulfiram during state 4 respiration, DPA did not further inhibit O2 uptake. Treating green bell peppers with DPA before storage did not alter the induction and abundance of the alternative oxidase protein in mitochondria which was maximally induced in peppers stored at 4 °C. Whether added before or after the uncoupler, 2,4-dinitrophenol, DPA negated the enhanced O2 uptake associated with uncoupling of electron transfer in isolated mitochondria. These observations indicate that DPA inhibits the flow of electrons through the cytochrome path, probably somewhere in the cytochrome bc 1 complex. Although the secondary amine function of DPA makes it a powerful antioxidant, the effectiveness of DPA in reducing chilling injury in green bell peppers and superficial scald in apples [Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf.] also may be due, in part, to its inhibition of respiration.
Abstract
Mitochondrial suspensions isolated from ‘Homestead’ tomatoes swelled on exposure to CCl4 or C2H4 and contracted upon the addition of ATP, Mg++, and BSA. The responses to C2H4 treatment were less evident than for the CCl4 treatment. Rates of swelling and contraction were more pronounced when the fruits were irradiated at the mature green or the 2-day breaker stages of maturity and during subsequent storage of these fruits.