Coffea canephora genotypes adopt distinct strategies to cope with drought and rehydration. We hypothesized that the greater drought tolerance of genotype ‘3V’ compared to ‘A1’, previously reflected in physiological and anatomical leaf traits after two water-stress (WS) cycles, could also be observed in P–V curve responses, root and branch anatomy, leaf midrib elongation (CVL), and root distribution. The ‘3V’ and ‘A1’ plants were grown under well-watered (WW) conditions and two cycles of water stress (WS). The ‘3V’ was more sensitive to WS, with reduced branch xylem vessel density (BXVD), while ‘A1’ demonstrated increased BXVD. Root xylem vessel area (RXVA) decreased to a greater extent in ‘3V’ than in ‘A1’, and both genotypes showed increased bulk elastic modulus. Regardless of water conditions, ‘A1’ maintained a higher relative leaf water content at the turgor loss point (RWC_TLP). Morphological acclimation did not occur in the second WS cycle. The ‘3V’ plants developed greater root mass in deeper soil layers than ‘A1’ under the WS condition. These findings suggest that ‘A1’ follows a conservative drought-avoidance strategy with lower physio-morphological plasticity, while ‘3V’ exhibits greater drought tolerance. Such responses highlighted coordinated physiological, morphological, and anatomical adaptations of the above- and below-ground organs for resource acquisition and conservation under WS.

Understanding the dynamics of physiological changes involved in the acclimation responses of plants after their exposure to repeated cycles of water stress is crucial to selecting resilient genotypes for regions with recurrent drought episodes. Under such background, we tried to respond to questions as: (1) Are there differences in the stomatal-related and non-stomatal responses during water stress cycles in different clones of Coffea canephora Pierre ex A. Froehner? (2) Do these C. canephora clones show a different response in each of the two sequential water stress events? (3) Is one previous drought stress event sufficient to induce a kind of “memory” in C. canephora? Seven-month-old plants of two clones (’3V’ and ‘A1’, previously characterized as deeper and lesser deep root growth, respectively) were maintained well-watered (WW) or fully withholding the irrigation, inducing soil water stress (WS) until the soil matric water potential (Ψ_msoil) reached ≅ −0.5 MPa (−500 kPa) at a soil depth of 500 mm. Two sequential drought events (drought-1 and drought-2) attained this Ψ_msoil after 19 days and were followed by soil rewatering until a complete recovery of leaf net CO₂ assimilation rate (A_net) during the recovery-1 and recovery-2 events. The leaf gas exchange, chlorophyll a fluorescence, and leaf reflectance parameters were measured in six-day frequency, while the leaf anatomy was examined only at the end of the second drought cycle. In both drought events, the WS plants showed reduction in stomatal conductance and leaf transpiration. The reduction in internal CO₂ diffusion was observed in the second drought cycle, expressed by increased thickness of spongy parenchyma in both clones. Those stomatal and anatomical traits impacted decreasing the A_net in both drought events. The ‘3V’ was less influenced by water stress than the ‘A1’ genotype in A_net, effective quantum yield in PSII photochemistry, photochemical quenching, linear electron transport rate, and photochemical reflectance index during the drought-1, but during the drought-2 event such an advantage disappeared. Such physiological genotype differences were supported by the medium xylem vessel area diminished only in ‘3V’ under WS. In both drought cycles, the recovery of all observed stomatal and non-stomatal responses was usually complete after 12 days of rewatering. The absence of photochemical impacts, namely in the maximum quantum yield of primary photochemical reactions, photosynthetic performance index, and density of reaction centers capable of Q_A reduction during the drought-2 event, might result from an acclimation response of the clones to WS. In the second drought cycle, the plants showed some improved responses to stress, suggesting “memory” effects as drought acclimation at a recurrent drought.

Coffea canephora cultivation areas in Brazil are frequently exposed to successive cycles of water deficit, triggering plant stress responses. In addition to water deficit, increased air temperature can act as a second stress factor. The recurrence of these stress factors may induce plant tolerance mechanisms, potentially mitigating future stress responses even of a different stress nature. We hypothesized that repeated cycles of water deficit can trigger tolerance mechanisms that make C. canephora leaves more resilient to supra-optimal temperatures. To test this hypothesis, young C. canephora plants were grown under non-limited water conditions for seven months (Ψ_mSoil > −20 kPa), after which they were subjected to two consecutive cycles of water deficit (Ψ_mSoil < −300 kPa), followed by rehydration. Two clones were used, ‘A1’ and ‘3V’, previously classified as drought sensitive and tolerant, respectively, considering the dynamics of physiological and architectural responses. After the second cycle, leaf discs were collected from completely expanded leaves formed during the two stress cycles and exposed to heat treatments (35 °C, 40 °C, 45 °C, 50 °C, and 55 °C) for 15 min in a water bath. Chlorophyll a fluorescence emission was then monitored, and the results were analyzed using OJIP transient kinetics and the JIP_Test. High temperatures induced negative changes in both OJIP kinetics and JIP_Test-derived parameters. A significant increase in F₀ and a reduction in F_M were observed mainly at 50 °C and 55 °C, due to changes in the stages of the OJIP curve. These changes impacted the “energy connectivity” and consequently the electron transport along the electron transfer chain (ETC), increasing energy dissipation, as confirmed by the JIP_Test variables. Despite the high temperature impacts, previous water deficit induced heat tolerance in clone ‘A1’, while it increased sensitivity in clone ‘3V’. This study suggests that selecting drought-resistant varieties should consider their subsequent response to short high-temperature stress to avoid cross-sensitivity caused by selecting for a single environmental factor.