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Coffee (Coffea spp.), a globally traded commodity, is a slow -growing tropical tree species that displays an improved photosynthetic performance when grown under elevated atmospheric CO2 concentrations ([CO2]). To investigate the mechanisms underlying this response, two commercial coffee cultivars (Catuaí and Obatã) were grown using the first free-air CO2 enrichment (FACE) facility in Latin America. Measurements were conducted in two contrasting growth seasons, which were characterized by the high (February) and low (August) sink demand. Elevated [CO2] led to increases in net photosynthetic rates (A) in parallel with decreased photorespiration rates, with no photochemical limitations to A. The stimulation of A by elevated CO2 supply was more prominent in August (56% on average) than in February (40% on average). Overall, the stomatal and mesophyll conductances, as well as the leaf nitrogen and phosphorus concentrations, were unresponsive to the treatments. Photosynthesis was strongly limited by diffusional constraints, particularly at the stomata level, and this pattern was little, if at all, affected by elevated [CO2]. Relative to February, starch pools (but not soluble sugars) increased remarkably (>500%) in August, with no detectable alteration in the maximum carboxylation capacity estimated on a chloroplast [CO2] basis. Upregulation of A by elevated [CO2] took place with no signs of photosynthetic downregulation, even during the period of low sink demand, when acclimation would be expected to be greatest.

Over the last decades, most information on the mechanisms underlying tolerance to drought has been gained by considering this stress as a single event that happens just once in the life of a plant, in contrast to what occurs under natural conditions where recurrent drought episodes are the rule. Here we explored mechanisms of drought toler- ance in coffee (Coffea canephora) plants from a broader perspective, integrating key aspects of plant physiology and biochemistry. We show that plants exposed to multiple drought events displayed higher photosynthetic rates, which were largely accounted for by biochemical rather than diffusive or hydraulic factors, than those submitted to drought for the first time. Indeed, these plants displayed higher activities of RuBisCO and other enzymes associated with car- bon and antioxidant metabolism. Acclimation to multiple drought events involved the expression of trainable genes related to drought tolerance and was also associated with a deep metabolite reprogramming with concordant altera- tions in central metabolic processes such as respiration and photorespiration. Our results demonstrate that plants exposed to multiple drought cycles can develop a differential acclimation that potentiates their defence mechanisms, allowing them to be kept in an ‘alert state’ to successfully cope with further drought events.

Plant defense compounds occur in floral nectar, but their ecological role is not well understood. We provide evidence that plant compounds pharmacologically alter pollinator behavior by enhancing their memory of reward. Honeybees rewarded with caffeine, which occurs naturally in nectar of Coffea and Citrus species, were three times as likely to remember a learned floral scent as were honeybees rewarded with sucrose alone. Caffeine potentiated responses of mushroom body neurons involved in olfactory learning and memory by acting as an adenosine receptor antagonist. Caffeine concentrations in nectar did not exceed the bees' bitter taste threshold, implying that pollinators impose selection for nectar that is pharmacologically active but not repellent. By using a drug to enhance memories of reward, plants secure pollinator fidelity and improve reproductive success.

Wild animals substantially support crop production by providing ecosystem services, such as pollination and natural pest control. However, the strengths of synergies between ecosystem services and their dependencies on land-use management are largely unknown. Here, we took an experimental approach to test the impact of land-use intensification on both individual and combined pollination and pest control services in coffee production systems at Mount Kilimanjaro. We established a full-factorial pollinator and vertebrate exclosure experiment along a land-use gradient from traditional homegardens (agroforestry systems), shaded coffee plantations to sun coffee plantations (total sample size = 180 coffee bushes). The exclusion of vertebrates led to a reduction in fruit set of ca 9%. Pollinators did not affect fruit set, but significantly increased fruit weight of coffee by an average of 7.4%. We found no significant decline of these ecosystem services along the land-use gradient. Pest control and pollination service were thus complementary, contributing to coffee production by affecting the quantity and quality of a major tropical cash crop across different coffee production systems at Mount Kilimanjaro.

Main conclusion The application of gibberellin and abscisic acid in coffee plants resulted in increased floral bud formation and fruit production by regulating key genes involved in flowering and hormonal biosynthesis pathways. Despite ongoing efforts, understanding hormonal regulation in perennial and woody species with complex phenological cycles, such as Coffea arabica L., remains limited. Given the global importance of coffee, identifying the main regulators of reproductive development is crucial to guarantee production, especially in face of climate change. This study investigated the effects of gibberellin (GA) and abscisic acid (ABA) at different concentrations (5, 25 and 100 ppm) in the reproductive development of C. arabica . Phenological analyses, molecular identification of genes involved in GA and ABA biosynthesis, degradation, and signaling, as well as gene expression profiling in leaves and floral buds during floral induction and development, were conducted. Promoter analysis of CaFT , quantification of 1-aminocyclopropane-1-carboxylate (ACC), enzymatic activity of ACC oxidase (ACO), and ethylene content were also assessed. Results showed that GA irrespective of concentration and ABA at 25 ppm applied during the main period of floral induction (March) significantly increased the number of floral buds, with ABA also accelerating the development. Similarly, applying these regulators in plants with floral buds at more advanced stages (August) increased the number of floral buds and fruit production in the GA (5 and 100 ppm) and ABA (25 and 100 ppm) treatments. Phylogenetic and molecular analyses identified genes related to GA and ABA biosynthesis, degradation, and signaling in coffee plants. GA and ABA treatments affected the expression of genes related to floral induction and organ formation, such as CaDELLA in March, which may relate to the increased number of floral buds. Moreover, in August, plants treated with 5 and 100 ppm GA and 100 ppm ABA showed up-regulation of CaFT1 expression, likely due to the down-regulation of CaCO during this period. In addition to GA-ABA interactions, our results suggest that GA promotes ACC accumulation in leaves in August, which may act as a mobile signal transported to floral buds, where its conversion to ethylene could regulate anthesis, highlighting a GA-ACC-ethylene interaction in coffee flowering. However, no significant differences in ethylene biosynthesis were observed in March with the application of these hormones, underscoring the incipient role of ethylene during floral induction in coffee. These results suggest reciprocal regulation of floral development by GA-ABA pathways in a dose-dependent manner and interacting with other hormonal pathways such as the ethylene biosynthesis in leaves and floral buds. These findings provide new insights into the hormonal regulation of coffee flowering, guiding field practices and breeding programs to maximize coffee production.

The understanding of acclimation strategies to low temperature and water availability is decisive to ensure coffee crop sustainability, since these environmental conditions determine the suitability of cultivation areas. In this context, the impacts of single and combined exposure to drought and cold were evaluated in three genotypes of the two major cropped species, Coffea arabica cv. Icatu, Coffea canephora cv. Apoatã, and the hybrid Obatã. Crucial traits of plant resilience to environmental stresses have been examined: photosynthesis, lipoperoxidation and the antioxidant response. Drought and/or cold promoted leaf dehydration, which was accompanied by stomatal and mesophyll limitations that impaired leaf C-assimilation in all genotypes. However, Icatu showed a lower impact upon stress exposure and a faster and complete photosynthetic recovery. Although lipoperoxidation was increased by drought (Icatu) and cold (all genotypes), it was greatly reduced by stress interaction, especially in Icatu. In fact, although the antioxidative system was reinforced under single drought and cold exposure (e.g., activity of enzymes as Cu,Zn-superoxide dismutase, ascorbate peroxidase, APX, glutathione reductase and catalase, CAT), the stronger increases were observed upon the simultaneous exposure to both stresses, which was accompanied with a transcriptional response of some genes, namely related to APX. Complementary, non-enzyme antioxidant molecules were promoted mostly by cold and the stress interaction, including α-tocopherol (in C. arabica plants), ascorbate (ASC), zeaxanthin, and phenolic compounds (all genotypes). In general, drought promoted antioxidant enzymes activity, whereas cold enhanced the synthesis of both enzyme and non-enzyme antioxidants, the latter likely related to a higher need of antioxidative capability when enzyme reactions were probably quite repressed by low temperature. Icatu showed the wider antioxidative capability, with the triggering of all studied antioxidative molecules by drought (except CAT), cold, and, particularly, stress interaction (except ASC), revealing a clear stress cross-tolerance. This justified the lower impacts on membrane lipoperoxidation and photosynthetic capacity under stress interaction conditions, related to a better ROS control. These findings are also relevant to coffee water management, showing that watering in the cold season should be largely avoided.

Robusta coffee (Coffea canephora ) accounts for about 80% of Uganda’s coffee production and supports over 2.2 million livelihoods. Yet recurrent droughts and erratic rainfall, exacerbated by climate change, pose severe threat to its productivity. Despite this threat, Ugandan breeding programs lack drought-tolerant varieties, largely because selection has not incorporated precise physiological traits linked to drought adaptation. Here, we evaluated 165 diverse Ugandan C. canephora genotypes, including local wild accessions, commercial lines, and breeders’ selections, under controlled screenhouse drought assay. Morphophysiological traits were collected before, during, and after drought stress. Using linear mixed-effects models, K-means clustering, and the Drought Factor Index, variation in drought tolerance was characterized across genotypes differing in biomass. Drought reduced water relations, gas exchange (gs and E), and PSII efficiency (Fv/Fm, Fo/Fv, ETR, Y(II), and PI) across all biomass groups. Nevertheless, 24 genotypes maintained higher DFI values, stable PSII function (Fv/Fm, PI, Fv/F₀), and less negative pre‑dawn water potential under severe stress. Among all traits, photosynthetic performance index (PI) and Fv/F₀ emerged as the most robust and biologically interpretable predictors of drought tolerance. These fluorescence‑based markers, together with the identified drought‑tolerant genotypes, provide a powerful foundation for accelerating climate‑smart coffee breeding in Uganda.

The advancement of digital agriculture combined with computational tools and Unmanned Aerial Vehicles (UAVs) has opened the way to large-scale data collection for the calculation of vegetation indices (VIs). These vegetation indexes (VIs) are useful for agricultural monitoring, as they highlight the inherent characteristics of vegetation and optimize the spatial and temporal evaluation of different crops. The experiment tested three coffee genotypes (Catuaí 62, E237 and Iapar 59) under five water regimes: (1) FI 100 (year-round irrigation with 100% replacement of evapotranspiration loss), (2) FI 50 (year-round irrigation with 50% evapotranspiration replacement), (3) WD 100 (no irrigation from June to September (dry season) and, thereafter, 100% evapotranspiration replacement), (4) WD 50 (no irrigation from June to September (water stress) and, thereafter, 50% evapotranspiration replacement) and (5) rainfed (no irrigation during the year). The irrigated treatments were watered with irrigation and precipitation. Most indices were highest in response to full irrigation (FI 100). The values of the NDVI ranged from 0.87 to 0.58 and the SAVI from 0.65 to 0.38, and the values of these indices were lowest for genotype E237 in the rainfed areas. The indices NDVI, OSAVI, MCARI, NDRE and GDVI were positively correlated—very strongly with photosynthesis (A) and strongly with transpiration (E) of the coffee trees. On the other hand, temperature-based indices, such as canopy temperature and the TCARI index correlated negatively with A, E and stomatal conductance (gs). Under full irrigation, the tested genotypes did not differ between the years of evaluation. Overall, the index values of Iapar 59 exceeded those of the other genotypes. The use of VIs to evaluate coffee tree performance under different water managements proved efficient in discriminating the best genotypes and optimal water conditions for each genotype. Given the economic importance of coffee as a crop and its susceptibility to extreme events such as drought, this study provides insights that facilitate the optimization of productivity and resilience of plantations under variable climatic conditions.

It has long been held that the low photosynthetic rates (A) of coffee leaves are largely associated with diffusive constraints to photosynthesis. However, the relative limitations of the stomata and mesophyll to the overall diffusional constraints to photosynthesis, as well as the coordination of leaf hydraulics with photosynthetic limitations, remain to be fully elucidated in coffee. Whether the low actual A under ambient CO 2 concentrations is associated with the kinetic properties of Rubisco and high (photo)respiration rates also remains elusive. Here, we provide a holistic analysis to understand the causes associated with low A by measuring a variety of key anatomical/hydraulic and photosynthetic traits in sun- and shade- grown coffee plants. We demonstrate that leaf hydraulic architecture imposes a major constraint on the maximisation of the photosynthetic gas exchange of coffee leaves. Regardless of the light treatments, A was mainly limited by stomatal factors followed by similar limitations associated with the mesophyll and biochemical constraints. No evidence of an inefficient Rubisco was found; rather, we propose that coffee Rubisco is well tuned for operating at low chloroplastic CO 2 concentrations. Finally, we contend that large diffusive resistance should lead to large CO 2 drawdown from the intercellular airspaces to the sites of carboxylation, thus favouring the occurrence of relatively high photorespiration rates, which ultimately leads to further limitations to A.

Integrative analysis of developing coffee seeds reveals the key cellular, metabolic, and regulatory processes associated with acquisition of desiccation tolerance in intermediate seeds Abstract The 'intermediate seed' category was defined in the early 1990s using coffee (Coffea arabica) as a model. In contrast to orthodox seeds, intermediate seeds cannot survive complete drying, which is a major constraint for seed storage and has implications for both biodiversity conservation and agricultural purposes. However, intermediate seeds are considerably more tolerant to drying than recalcitrant seeds, which are highly sensitive to desiccation. To gain insight into the mechanisms governing such differences, changes in desiccation tolerance (DT), hormone contents, and the transcriptome were analysed in developing coffee seeds. Acquisition of DT coincided with a dramatic transcriptional switch characterised by the repression of primary metabolism, photosynthesis, and respiration, and the up-regulation of genes coding for late-embryogenesis abundant (LEA) proteins, heat-shock proteins (HSPs), and antioxidant enzymes. Analysis of the heat-stable proteome in mature coffee seeds confirmed the accumulation of LEA proteins identified at the transcript level. Transcriptome analysis also suggested a major role for ABA and for the transcription factors CaHSFA9, CaDREB2G, CaANAC029, CaPLATZ, and CaDOG-like in DT acquisition. The ability of CaHSFA9 and CaDREB2G to trigger HSP gene transcription was validated by Agrobacterium-mediated transformation of coffee somatic embryos.