Jorge Marques da Silva

Although the use of the term “sentience” in philosophy dates to the 17th century, its use expanded particularly in the second half of the 20th century, with the emergence of non-anthropocentric ethics. A search for sentience in the animal kingdom began, which required the identification of a set of evidential sources. The difficulty in establishing, beyond any reasonable doubt, the existence of sentience in animals more distantly related to humans, and the consequences that the matter has for the legislation on animal protection, led to the proposal to apply the precautionary principle in animal welfare policy. This principle establishes that the fulfilment of minimum criteria is enough for the existence of sentience to be admitted. This led to the attribution of sentience to invertebrates, namely molluscs, insects, and crustaceans. As a result, the possibility of including decapod crustaceans into animal legislation is currently discussed. In this article, we will perform the comparative exercise of applying a framework of minimum criteria of sentience, previously applied to crustaceans of the order Decapoda, to vascular plants. The framework is composed of eight independent criteria and the likelihood of plants meeting each of them will be assessed and discussed in the light of the existing empirical evidence.

Introduction: Artemisinin is a secondary metabolite well-known for its use in the treatment of malaria. It also displays other antimicrobial activities which further increase its interest. At present, Artemisia annua is the sole commercial source of the substance, and its production is limited, leading to a global deficit in supply. Furthermore, the cultivation of A. annua is being threatened by climate change. Specifically, drought stress is a major concern for plant development and productivity, but, on the other hand, moderate stress levels can elicit the production of secondary metabolites, with a putative synergistic interaction with elicitors such as chitosan oligosaccharides (COS). Therefore, the development of strategies to increase yield has prompted much interest. With this aim, the effects on artemisinin production under drought stress and treatment with COS, as well as physiological changes in A. annua plants are presented in this study.Methods: Plants were separated into two groups, well-watered (WW) and drought-stressed (DS) plants, and in each group, four concentrations of COS were applied (0, 50,100 and 200 mg•L-1). Afterwards, water stress was imposed by withholding irrigation for 9 days.Results: Therefore, when A. annua was well watered, COS did not improve plant growth, and the upregulation of antioxidant enzymes hindered the production of artemisinin. On the other hand, during drought stress, COS treatment did not alleviate the decline in growth at any concentration tested. However, higher doses improved the water status since leaf water potential (YL) improved by 50.64% and relative water content (RWC) by 33.84% compared to DS plants without COS treatment. Moreover, the combination of COS and drought stress caused damage to the plant’s antioxidant enzyme defence, particularly APX and GR, and reduced the amount of phenols and flavonoids. This resulted in increased ROS production and enhanced artemisinin content by 34.40% in DS plants treated with 200 mg•L-1 COS, compared to control plants.Conclusion: These findings underscore the critical role of ROS in artemisinin biosynthesis and suggest that COS treatment may boost artemisinin yield in crop production, even under drought conditions.

MicroRNAs (miRNAs) are non-coding small RNAs that play crucial roles in plant development and stress responses and can regulate plant interactions with beneficial soil microorganisms such as arbuscular mycorrhizal fungi (AMF). To determine if root inoculation with distinct AMF species affected miRNA expression in grapevines subjected to high temperatures, RNA-seq was conducted in leaves of grapevines inoculated with either Rhizoglomus irregulare or Funneliformis mosseae and exposed to a high-temperature treatment (HTT) of 40 °C for 4 h per day for one week. Our results showed that mycorrhizal inoculation resulted in a better plant physiological response to HTT. Amongst the 195 identified miRNAs, 83 were considered isomiRs, suggesting that isomiRs can be biologically functional in plants. The number of differentially expressed miRNAs between temperatures was higher in mycorrhizal (28) than in non-inoculated plants (17). Several miR396 family members, which target homeobox-leucine zipper proteins, were only upregulated by HTT in mycorrhizal plants. Predicted targets of HTT-induced miRNAs in mycorrhizal plants queried to STRING DB formed networks for Cox complex, and growth and stress-related transcription factors such as SQUAMOSA promoter-binding-like-proteins, homeobox-leucine zipper proteins and auxin receptors. A further cluster related to DNA polymerase was found in R. irregulare inoculated plants. The results presented herein provide new insights into miRNA regulation in mycorrhizal grapevines under heat stress and can be the basis for functional studies of plant-AMF-stress interactions.

The ubiquitin-like modifying peptide SMALL UBIQUITIN-LIKE MODIFIER (SUMO) has become a known modulator of the plant response to multiple environmental stimuli. A common feature of many of these external stresses is the production of reactive oxygen species (ROS). Taking into account that SUMO conjugates rapidly accumulate in response to an external oxidative stimulus, it is likely that ROS and sumoylation converge at the molecular and regulatory levels. In this study, we explored the SUMO–ROS relationship, using as a model the Arabidopsis (Arabidopsis thaliana) null mutant of the major SUMO-conjugation enhancer, the E3 ligase SAP AND MIZ 1 (SIZ1). We showed that SIZ1 is involved in SUMO conjugate increase when primed with both exogenous and endogenous ROS. In siz1, seedlings were sensitive to oxidative stress imposition, and mutants accumulated different ROS throughout development. We demonstrated that the deregulation in hydrogen peroxide and superoxide homeostasis, but not of singlet O₂ (¹O₂), was partially due to SA accumulation in siz1. Furthermore, transcriptomic analysis highlighted a transcriptional signature that implicated siz1 with ¹O₂ homeostasis. Subsequently, we observed that siz1 displayed chloroplast morphological defects and altered energy dissipation activity and established a link between the chlorophyll precursor protochlorophyllide and deregulation of PROTOCHLOROPHYLLIDE OXIDOREDUCTASE A (PORA), which is known to drive overproduction of ¹O₂. Ultimately, network analysis uncovered known and additional associations between transcriptional control of PORA and SIZ1-dependent sumoylation. Our study connects sumoylation, and specifically SIZ1, to the control of chloroplast functions and places sumoylation as a molecular mechanism involved in ROS homeostatic and signaling events.

As a result of the development of non-invasive optical spectroscopy, the number of prospective technologies of plant monitoring is growing. Being implemented in devices with different functions and hardware, these technologies are increasingly using the most advanced data processing algorithms, including machine learning and more available computing power each time. Optical spectroscopy is widely used to evaluate plant tissues, diagnose crops, and study the response of plants to biotic and abiotic stress. Spectral methods can also assist in remote and non-invasive assessment of the physiology of photosynthetic biofilms and the impact of plant species on biodiversity and ecosystem stability. The emergence of high-throughput technologies for plant phenotyping and the accompanying need for methods for rapid and non-contact assessment of plant productivity has generated renewed interest in the application of optical spectroscopy in fundamental plant sciences and agriculture. In this perspective paper, starting with a brief overview of the scientific and technological backgrounds of optical spectroscopy and current mainstream techniques and applications, we foresee the future development of this family of optical spectroscopic methodologies.

Plant phenotyping is an emerging science that combines multiple methodologies and protocols to measure plant traits (e.g., growth, morphology, architecture, function, and composition) at multiple scales of organization. Manual phenotyping remains as a major bottleneck to the advance of plant and crop breeding. Such constraint fostered the development of high throughput plant phenotyping (HTPP), which is largely based on imaging approaches and automatized data retrieval and processing. Field phenotyping still poses major challenges and the progress of HTPP for field conditions can be relevant to support selection and breeding of grapevine. The aim of this review is to discuss potential and current methods to improve field phenotyping of grapevine to support characterization of inter- and intravarietal diversity. Vitis vinifera has a large genetic diversity that needs characterization, and the availability of methods to support selection of plant material (polyclonal or clonal) able to withstand abiotic stress is paramount. Besides being time consuming, complex and expensive, field experiments are also affected by heterogeneous and uncontrolled climate and soil conditions, mostly due to the large areas of the trials and to the high number of traits to be observed in a number of individuals ranging from hundreds to thousands. Therefore, adequate field experimental design and data gathering methodologies are crucial to obtain reliable data. Some of the major challenges posed to grapevine selection programs for tolerance to water and heat stress are described herein. Useful traits for selection and related field phenotyping methodologies are described and their adequacy for large scale screening is discussed.

Increasing temperatures and extended drought episodes are among the major constraints affecting food production. Maize has a relatively high temperature optimum for photosynthesis compared to C₃ crops, however, the response of this important C₄ crop to the combination of heat and drought stress is poorly understood. Here, we hypothesized that resilience to high temperature combined with water deficit (WD) would require efficient regulation of the photosynthetic traits of maize, including the C₄–CO₂ concentrating mechanism (CCM). Two genotypes of maize with contrasting levels of drought and heat tolerance, B73 and P0023, were acclimatized at high temperature (38°C versus 25°C) under well-watered (WW) or WD conditions. The photosynthetic performance was evaluated by gas exchange and chlorophyll a fluorescence, and in vitro activities of key enzymes for carboxylation (phosphoenolpyruvate carboxylase), decarboxylation (NADP-malic enzyme), and carbon fixation (Rubisco). Both genotypes successfully acclimatized to the high temperature, although with different mechanisms: while B73 maintained the photosynthetic rates by increasing stomatal conductance (gs), P0023 maintained gs and showed limited transpiration. When WD was experienced in combination with high temperatures, limited transpiration allowed water-savings and acted as a drought stress avoidance mechanism. The photosynthetic efficiency in P0023 was sustained by higher phosphorylated PEPC and electron transport rate (ETR) near vascular tissues, supplying chemical energy for an effective CCM. These results suggest that the key traits for drought and heat tolerance in maize are limited transpiration rate, allied with a synchronized regulation of the carbon assimilation metabolism. These findings can be exploited in future breeding efforts aimed at improving maize resilience to climate change.

Plant breeders and plant physiologists are deeply committed to high throughput plant phenotyping for drought tolerance. A combination of artificial intelligence with reflectance spectroscopy was tested, as a non-invasive method, for the automatic classification of plant drought stress. Arabidopsis thaliana plants (ecotype Col-0) were subjected to different levels of slowly imposed dehydration (S0, control; S1, moderate stress; S2, severe stress). The reflectance spectra of fully expanded leaves were recorded with an Ocean Optics USB4000 spectrometer and the soil water content (SWC, %) of each pot was determined. The entire data set of the reflectance spectra (intensity vs. wavelength) was given to different machine learning (ML) algorithms, namely decision trees, random forests and extreme gradient boosting. The performance of different methods in classifying the plants in one of the three drought stress classes (S0, S1 and S2) was measured and compared. All algorithms produced very high evaluation scores (F1 > 90%) and agree on the features with the highest discriminative power (reflectance at ~670 nm). Random forests was the best performing method and the most robust to random sampling of training data, with an average F1-score of 0.96 ± 0.05. This classification method is a promising tool to detect plant physiological responses to drought using high-throughput pipelines.

Landraces are rightfully known as the backbone of sustainable food production, particularly in areas experiencing significant environmental constraints. However, protecting landraces from genetic erosion and valuing their potential for plant breeding and sustainable food production requires in-depth understanding of their phenotypic traits. Two Guinea-Bissau landraces (GA, GV) and two elite cultivars (P98438, P0023) were subjected to drought stress for 7 (S1) and 12 (S2) days. After this period plants were rewatered (R). The relative water content (RWC) was unchanged in S1 and decreased in S2 in all genotypes. Chlorophyll a fluorescence parameters changed moderately in S1 and remarkably in S2, including on GA and GV plants, where a decrease of Fv/Fm and PI, and an increase in absorbed, trapped, and dissipated energy per reaction center, was found. P0023 plants showed the most contrasting behavior to Guinea-Bissau genotypes, presenting an increase in Fv/Fm and PI values and a decrease in the specific energy fluxes per reaction center (RC), whereas P9838 presented an intermediate behavior. Drought (S1 and S2) decreased the amount of chlorophyll (Chl.) and carotenoids in GA and GV plants. On the contrary, in the P0023, the only pigment to decrease with stress was Chl. b. Fatty acid (FA) analyses allowed for the identification of C16:0, C18:2, and C18:3 in larger amounts, and C14:0, C16:1t, C18:0, and C18:1 acids in lower abundance. Drought stress decreased C18:3, the double bond index, and the total FA (except for P0023 and GV in S2) and increased C14:0 and C16:0 acids. The expression of phospholipase D (PLD) was higher at S2. After recovery, PLD expression presented a notorious decrease on the Guinea-Bissau landraces. P0023 showed the smallest alterations after recovery, while Guinea’s plants suffered more radical alterations leading to the conclusion that Guinea’s landraces were more drought-sensitive and that their recovery rate was impaired.