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The plant hormones brassinosteroids (BR) and ethylene (ET) act together to regulate plant metabolism. We used BR and 1-methylcyclopropene (1-MCP, an ET action inhibitor) to elucidate the interactions between both hormones for the regulation of mitochondrial respiratory pathways in papaya fruit. The exogenous application of the 24-epibrassinolide (epiBR) enhanced the alternative oxidase (AOX) capacity. While treatment with Brz2001 (Brz is a specific inhibitor of the BR synthesis) also enhanced AOX capacity, these effects lacked in fruit treated simultaneously with epiBR and Brz. Changing the BR level had no effect on ET emission rate in the first 24 h, but a reduction in ET emission was observed in Brz-treated fruit on the fifth day. Together with Brz, epiBR increased the ET production on the fifth day, following the day in which the treatment was carried out. When the ET sensitivity of fruit was inhibited by the application of 1-MCP, the effects of epiBR and Brz were opposite to those obtained without 1-MCP. AOX capacity was slightly inhibited by epiBR in fruit pre-treated with 1-MCP. Data suggest that BR and ET act antagonistically, therefore regulating, directly or indirectly, AOX capacity during papaya fruit ripening.

Enhanced respiration during ripening in climacteric fruits is sometimes associated with an uncoupling between the ATP synthesis and the mitochondrial electron transport chain. While the participation of two energy-dissipating systems, one of which is mediated by the alternative oxidase (AOX) and the other mediated by the uncoupling protein (UCP), has been linked to fruit ripening, the relation between the activation of both mitochondrial uncoupling systems with the transient increase of ethylene synthesis (ethylene peak) remains unclear. To elucidate this question, ethylene emission and the two uncoupling (AOX and UCP) pathways were monitored in harvested papaya fruit during the ripening, from green to fully yellow skin. The results confirmed the typical climacteric behavior for papaya fruit: an initial increase in endogenous ethylene emission which reaches a maximum (peak) in the intermediate ripening stage, before finally declining to a basal level in ripe fruit. Respiration of intact fruit also increased and achieved higher levels at the end of ripening. On the other hand, in purified mitochondria extracted from fruit pulp the total respiration and respiratory control decrease while an increase in the participation of AOX and UCP pathways was markedly evident during papaya ripening. There was an increase in the AOX capacity during the transition from green fruit to the intermediate stage that accompanied the transient ethylene peak, while the O2 consumption triggered by UCP activation increased by 80% from the beginning to end stage of fruit ripening. Expression analyses of AOX (AOX1 and 2) and UCP (UCP1-5) genes revealed that the increases in the AOX and UCP capacities were linked to a higher expression of AOX1 and UCP (mainly UCP1) genes, respectively. In silico promoter analyses of both genes showed the presence of ethylene-responsive cis-elements in UCP1 and UCP2 genes. Overall, the data suggest a differential activation of AOX and UCP pathways in regulation related to the ethylene peak and induction of specific genes such as AOX1 and UCP1.

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Antioxidant system is involved in arbuscular mycorrhizal symbiosis, but its role during the colonization process is still poorly understood. To gain new insights into the role of antioxidant system during root colonization by arbuscular mycorrhizal fungi, the activities of key antioxidant enzymes were evaluated in tomato (Solanum lycopersicum L.) roots inoculated with six strains of different genera and species: two Glomus mosseae, Glomus cubense, Glomus intraradices, Glomus sp. and Acaulospora scrobiculata. Glomus cubense and A. scrobiculata strains reached the highest infectivity levels with maximum values of colonization frequency and intensity of 29-10.88% and 18-9.20%, respectively; G. mosseae strains showed an intermediate infectivity, both with 15% of colonization frequency and maximum intensities of 7.64-7.06%, respectively; while the infectivity levels of Glomus sp. and G. intraradices strains were the lowest with colonization frequency-13% and intensities-5.07 and 5.41, respectively. Some activity patterns of peroxidase, superoxide dismutase, and polyphenol oxidase enzymes were not specific for early or late colonization stages neither for the colonization level and type of strain. However, a unique superoxide dismutase-band presents at early colonization and the low level of guaiacolperoxidase activity at later stages presents in all inoculated roots indicate that these antioxidant responses are independent of colonization degree and strain. Taking together, our data suggest that alterations of the antioxidant enzyme activities are not general characteristics of the colonization process by arbuscular mycorrhizal fungi, probably having the key role on those responses the specific feature of each strain rather than colonization per se.