Ajaree Thonglim

Numerous plant species can actively pressurize water in their xylem. It has been hypothesized that regular peaks in positive pressure help plants to remove xylem embolism and to replenish water reservoirs. This may happen after rewatering when plants have suffered from soil water deficit (WD) and/or following diurnal cycles under well-watered (WW) conditions. Using X-ray microcomputed tomography (µCT), we tested these hypotheses in tomato plants by quantifying xylem functionality and internal water pools in stems of 35 plants subjected to WD intensities before and after rewatering and 14 WW plants. Positive xylem pressure was detected in only 30% of the WD plants after rewatering, with a majority of this 30% showing no or rare events of xylem embolism repair. However, all WD tomato plants restored their internal water pools after rewatering, independently of their ability to generate detectable positive xylem pressure. In WW plants, positive xylem pressure was consistently generated and allowed for repair of the few embolized vessels. The ability of tomato plants to rehydrate non-xylary parenchyma tissues after WD and to repair embolisms under WW conditions sheds new light on alternative water-use mechanisms of herbaceous species to recover from drought.

Soil water deficit (WD) significantly impacts plant survival and crop yields. Many gaps remain in our understanding of the synergistic coordination between molecular and ecophysiological responses delaying substantial drought-induced effects on plant growth. To investigate this synergism in tomato leaves, we combined molecular, ecophysiological, and anatomical methods to examine gene expression patterns and physio-anatomical characteristics during a progressing WD experiment. Four sampling points were selected for transcriptomic analysis based on the key ecophysiological responses of the tomato leaves: 4 and 5 days after WD (d-WD), corresponding to 10% and 90% decrease in leaf stomatal conductance; 8 d-WD, the leaf wilting point; and 10 d-WD, when air embolism blocks 12% of the leaf xylem water transport. At 4 d-WD, upregulated genes were mostly linked to ABA-independent responses, with larger-scale ABA-dependent responses occurring at 5 d-WD. At 8 d-WD, we observed an upregulation of heat shock transcription factors, and two days later (10 d-WD), we found a strong upregulation of oxidative stress transcription factors. Finally, we found that young leaves present a stronger dehydration tolerance than mature leaves at the same drought intensity level, presumably because young leaves upregulate genes related to increased callose deposition resulting in limiting water loss to the phloem, and related to increased cell rigidity by modifying cell wall structures. This combined dataset will serve as a framework for future studies that aim to obtain a more holistic WD plant response at the molecular, ecophysiological and anatomical level.

Soil water deficit (WD) is one of the most important abiotic stresses affecting plant survival and crop yield. Despite its economic relevance, many gaps remain in our understanding of how crops respond to WD, especially concerning the synergistic coordination of molecular and ecophysiological adaptations delaying plant damage and mortality. In this study, we investigated the gene expression imposed by a progressive WD and combined it with measurements pointing to key ecophysiological thresholds in leaves of tomato plants. We uncovered the transcriptomic changes in mature leaves at four stages defined by physiological markers relating to different WD intensities: partial stomatal closure, complete stomatal closure, after leaf wilting, and beginning of embolism development in the veins. By identifying key transcription factors (TFs) across these progressively worsening WD stages, we investigated the timing and impact of ABA-(in)dependent gene regulatory pathways during WD. In addition, we compared the transcriptome in young developing versus mature leaves and explored the physiological mechanisms that may explain the higher tolerance to dehydration in younger leaves. By correlating the transcriptomic changes to precise ecophysiological measurements, the combined dataset will serve as a framework for future studies comparing leaf molecular and physiological responses to WD at specific intensities. Highlight Integrated ecophysiological and gene expression analyses identify key mechanisms underlying the different thresholds of tomato responses to water deficit

The synergy between drought-responsive traits across different organs is crucial in the whole-plant mechanism influencing drought resilience. These organ interactions, however, are poorly understood, limiting our understanding of drought response strategies at the whole-plant level. Therefore, we need more integrative studies, especially on herbaceous species that represent many important food crops but remain underexplored in their drought response. We investigated inflorescence stems and rosette leaves of six Arabidopsis thaliana genotypes with contrasting drought tolerance, and combined anatomical observations with hydraulic measurements and gene expression studies to assess differences in drought response. The soc1ful double mutant was the most drought-tolerant genotype based on its synergistic combination of low stomatal conductance, largest stomatal safety margin, more stable leaf water potential during non-watering, reduced transcript levels of drought stress marker genes, and reduced loss of chlorophyll content in leaves, in combination with stems showing the highest embolism resistance, most pronounced lignification, and thickest intervessel pit membranes. In contrast, the most sensitive Cvi ecotype shows the opposite extreme of the same set of traits. The remaining four genotypes show variations in this drought syndrome. Our results reveal that anatomical, ecophysiological, and molecular adaptations across organs are intertwined, and multiple (differentially combined) strategies can be applied to acquire a certain level of drought tolerance.

The ability to avoid drought-induced embolisms in the xylem is one of the essential traits for plants to survive periods of water shortage. Over the past three decades, hydraulic studies have been focusing on trees, which limits our ability to understand how herbs tolerate drought. Here we investigate the embolism resistance in inflorescence stems of four Arabidopsis thaliana accessions that differ in growth form and drought response. We assess functional traits underlying the variation in embolism resistance amongst the accessions studied using detailed anatomical observations. Vulnerability to xylem embolism was evaluated via vulnerability curves using the centrifuge technique and linked with detailed anatomical observations in stems using light microscopy and transmission electron microscopy. The data show significant differences in stem P_50, varying 2-fold from −1.58 MPa in the Cape Verde Island accession to −3.07 MPa in the woody soc1 ful double mutant. Out of all the anatomical traits measured, intervessel pit membrane thickness (T_PM) best explains the differences in P₅₀, as well as P₁₂ and P₈₈. The association between embolism resistance and T_PM can be functionally explained by the air-seeding hypothesis. There is no evidence that the correlation between increased woodiness and increased embolism resistance is directly related to functional aspects. However, we found that increased woodiness is strongly linked to other lignification characters, explaining why mechanical stem reinforcement is indirectly related to increased embolism resistance. The woodier or more lignified accessions are more resistant to embolism than the herbaceous accessions, confirming the link between increased stem lignification and increased embolism resistance, as also observed in other lineages. Intervessel pit membrane thickness and, to a lesser extent, theoretical vessel implosion resistance and vessel wall thickness are the missing functional links between stem lignification and embolism resistance.