Low expression of lipoxygenase 3 (LOX3) enhances the retention of kernel tocopherols in maize during storage.

BACKGROUND: Deficiency of vitamin E results in several neurological and age-related disorders in humans. Utilization of maize mutants with favourable vte4-allele led to the development of several α-tocopherol (vitamin E) rich (16-19 µg/g) maize hybrids worldwide. However, the degradation of tocopherols during post-harvest storage substantially affects the efficacy of these genotypes. METHODS AND RESULTS: We studied the role of lipoxygenase enzyme and Lipoxygenase 3 (LOX3) gene on the degradation of tocopherols at monthly intervals under traditional storage up to six months in two vte4-based contrasting-tocopherol retention maize inbreds viz. HKI323-PVE and HKI193-1-PVE. The analysis revealed significant degradation of tocopherols across storage intervals in both the inbreds. Lower retention of α-tocopherol was noticed in HKI193-1-PVE. HKI323-PVE with the higher retention of α-tocopherol showed lower lipoxygenase activity throughout the storage intervals. LOX3 gene expression was higher (~ 1.5-fold) in HKI193-1-PVE compared to HKI323-PVE across the storage intervals. Both lipoxygenase activity and LOX3 expression peaked at 120 days after storage (DAS) in both genotypes. Further, a similar trend was observed for LOX3 expression and lipoxygenase activity. The α-tocopherol exhibited a significantly negative correlation with lipoxygenase enzyme and expression of LOX3 across the storage intervals. CONCLUSIONS: HKI323-PVE with high tocopherol retention, low -lipoxygenase activity, and -LOX3 gene expression can act as a potential donor in the vitamin E biofortification program. Protein-protein association network analysis also indicated the independent effect of vte4 and LOX genes. This is the first comprehensive report analyzing the expression of the LOX3 gene and deciphering its vital role in the retention of α-tocopherol in biofortified maize varieties under traditional storage.

Contribution of Deep Learning in the Investigation of Possible Dual LOX-3 Inhibitors/DPPH Scavengers: The Case of Recently Synthesized Compounds.

Even though non-steroidal anti-inflammatory drugs are the most effective treatment for inflammatory conditions, they have been linked to negative side effects. A promising approach to mitigating potential risks, is the development of new compounds able to combine anti-inflammatory with antioxidant activity to enhance activity and reduce toxicity. The implication of reactive oxygen species in inflammatory conditions has been extensively studied, based on the pro-inflammatory properties of generated free radicals. Drugs with dual activity (i.e., inhibiting inflammation related enzymes, e.g., LOX-3 and scavenging free radicals, e.g., DPPH) could find various therapeutic applications, such as in cardiovascular or neurodegenerating disorders. The challenge we embarked on using deep learning was the creation of appropriate classification and regression models to discriminate pharmacological activity and selectivity as well as to discover future compounds with dual activity prior to synthesis. An accurate filter algorithm was established, based on knowledge from compounds already evaluated in vitro, that can separate compounds with low, moderate or high activity. In this study, we constructed a customized highly effective one dimensional convolutional neural network (CONV1D), with accuracy scores up to 95.2%, that was able to identify dual active compounds, being LOX-3 inhibitors and DPPH scavengers, as an indication of simultaneous anti-inflammatory and antioxidant activity. Additionally, we created a highly accurate regression model that predicted the exact value of effectiveness of a set of recently synthesized compounds with anti-inflammatory activity, scoring a root mean square error value of 0.8. Eventually, we succeeded in observing the manner in which those newly synthesized compounds differentiate from each other, regarding a specific pharmacological target, using deep learning algorithms.

Antisense suppression of LOX3 gene expression in rice endosperm enhances seed longevity.

Lipid peroxidation plays a major role in seed longevity and viability. In rice grains, lipid peroxidation is catalyzed by the enzyme lipoxygenase 3 (LOX3). Previous reports showed that grain from the rice variety DawDam in which the LOX3 gene was deleted had less stale flavour after grain storage than normal rice. The molecular mechanism by which LOX3 expression is regulated during endosperm development remains unclear. In this study, we expressed a LOX3 antisense construct in transgenic rice (Oryza sativa L.) plants to down-regulate LOX3 expression in rice endosperm. The transgenic plants exhibited a marked decrease in LOX mRNA levels, normal phenotypes and a normal life cycle. We showed that LOX3 activity and its ability to produce 9-hydroperoxyoctadecadienoic acid (9-HPOD) from linoleic acid were significantly lower in transgenic seeds than in wild-type seeds by measuring the ultraviolet absorption of 9-HPOD at 234 nm and by high-performance liquid chromatography. The suppression of LOX3 expression in rice endosperm increased grain storability. The germination rate of TS-91 (antisense LOX3 transgenic line) was much higher than the WT (29% higher after artificial ageing for 21 days, and 40% higher after natural ageing for 12 months). To our knowledge, this is the first report to demonstrate that decreased LOX3 expression can preserve rice grain quality during storage with no impact on grain yield, suggesting potential applications in agricultural production.

Four shades of detachment: regulation of floral organ abscission.

Abscission of floral organs from the main body of a plant is a dynamic process that is developmentally and environmentally regulated. In the past decade, genetic studies in Arabidopsis have identified key signaling components and revealed their interactions in the regulation of floral organ abscission. The phytohormones jasmonic acid (JA) and ethylene play critical roles in flower development and floral organ abscission. These hormones regulate the timing of floral organ abscission both independently and inter-dependently. Although significant progress has been made in understanding abscission signaling, there are still many unanswered questions. These include considering abscission in the context of reproductive development and interplay between hormones embedded in the developmental processes. This review summarizes recent advances in the identification of molecular components in Arabidopsis and discusses their relationship with reproductive development. The emerging roles of hormones in the regulation of floral organ abscission, particularly by JA and ethylene, are examined.