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Póster presentado en el XIM4 4th Xylem International Meeting Padua, 25-27 September 2019

Comunicación oral presentada en UC Davis Pressure Chamber Conference or 'Bombference', a conference on the use of the Scholander pressure chamber in plant research, 27 to 28 September 2018 Davis (California)

Plants have evolved a water transport system that relies on water sustaining a tensile force. Counter intuitively, this means water moves through the plant as a liquid under negative absolute pressures. This mechanism is made possible by the intricate plumbing system that constitutes the xylem tissue of plants. However, water under tension is prone to cavitation, which results in the formation of a gas bubble (embolism). Embolism reduces the capacity of the xylem tissue to deliver water to the canopy, eventually causing dieback and whole plant mortality. Xylem embolism is exacerbated by environmental stresses and is now considered one of the leading causes of plant mortality resulting from drought stress.Non-invasive imaging techniques offer the potential to make direct observations on intact plants at high resolution and in real time. In this presentation, I discuss recent exciting developments in the application of non-invasive imaging technologies such as X-ray Micro Computed Tomography (microCT) and optical imaging to studies of plant vascular function. This includes visualisation of xylem networks during drought stress and recovery in leaves, stems and roots. MicroCT imaging of stems and roots indicated that significant embolism formation occurs at similar time points and levels of water stress in dehydrating plants. This result was observed in herbaceous and woody species, and is surprising given previous hydraulic measurements indicating that, within a plant, roots were more vulnerable to drought-induced embolism than stems. A newly developed optical technique indicates that leaf vasculature is also similar in vulnerability to stems and roots. The overlap in vulnerability suggests that induction of embolism occurs at the same time in different organs or is propagated rapidly through the plant. In examining recovery from drought stress, we saw little evidence of embolism refilling in the xylem of woody plants, except in cases where substantial root pressure is produced. These results suggest that embolism refilling is less widespread than previously thought.

25 páginas.- 8 figuras.- 2 tablas.- 66 referencias.- Additional supporting information can be found online in the Supporting Information section at the end of this article., eaf water potential (psi(leaf)), typically measured using the pressure chamber, is the most important metric of plant water status, providing high theoretical value and information content for multiple applications in quantifying critical physiological processes including drought responses. Pressure chamber measurements of psi(leaf) (psi(leafPC)) are most typical, yet, the practical complexity of the technique and of the underlying theory has led to ambiguous understanding of the conditions to optimize measurements. Consequently, specific techniques and precautions diversified across the global research community, raising questions of reliability and repeatability. Here, we surveyed specific methods of psi(leafPC) from multiple laboratories, and synthesized experiments testing common assumptions and practices in psi(leafPC) for diverse species: (i) the need for equilibration of previously transpiring leaves; (ii) leaf storage before measurement; (iii) the equilibration of psi(leaf) for leaves on bagged branches of a range of dehydration; (iv) the equilibration of psi(leaf) across the lamina for bagged leaves, and the accuracy of measuring leaves with artificially 'elongated petioles'; (v) the need in psi(leaf) measurements for bagging leaves and high humidity within the chamber; (vi) the need to avoid liquid water on leaf surfaces; (vii) the use of 'pulse' pressurization versus gradual pressurization; and (viii) variation among experimenters in psi(leafPC) determination. Based on our findings we provide a best practice protocol to maximise accuracy, and provide recommendations for ongoing species-specific tests of important assumptions in future studies., European Union's Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No. 751918-AgroPHYS; Marie Curie Fellowship (FP7-PEOPLE-2013-IOF-624473); Austrian research agency (FWF) project P32203; National Science Foundation (Grant IOS-#1457279)

13 páginas.- 7 figuras.- 92 referencias, The mechanisms by which woody plants recover xylem hydraulic capacity after drought stress are not well understood, particularly with regard to the role of embolism refilling. We evaluated the recovery of xylem hydraulic capacity in young Eucalyptus saligna plants exposed to cycles of drought stress and rewatering. Plants were exposed to moderate and severe drought stress treatments, with recovery monitored at time intervals from 24 h to 6 months after rewatering. The percentage loss of xylem vessels due to embolism (PLV) was quantified at each time point using microcomputed tomography with stem water potential (Ψx) and canopy transpiration (Ec) measured before scans. Plants exposed to severe drought stress suffered high levels of embolism (47.38% ± 10.97% PLV) and almost complete canopy loss. No evidence of embolism refilling was observed at 24 h, 1 week, or 3 weeks after rewatering despite rapid recovery in Ψx. Recovery of hydraulic capacity was achieved over a 6-month period by growth of new xylem tissue, with canopy leaf area and Ec recovering over the same period. These findings indicate that E. saligna recovers slowly from severe drought stress, with potential for embolism to persist in the xylem for many months after rainfall events. © 2022 John Wiley & Sons Ltd., This study was supported by an ARC Discovery Project (DP170100761) to BC and TJB and an ARC Future Fellowship (FT130101115) to BC. BM acknowledges support from the ARC Laureate Fellowship FL190100003. CMR‐D was supported by an Individual Fellowship from the European Union's Horizon 2020 research and innovation program under the Marie Skłodowska‐Curie grant agreement no. 751918‐AgroPHYS. JMRP was supported by the ORNL, managed by UT‐Battelle, LLC, for the DOE under contract DE‐AC05‐1008 00OR22725.

14 páginas.- 6 figuras.- referencias.- Additional Supporting Information may be found online in the Supporting Information section at the end of the article https://nph.onlinelibrary.wiley.com/action/downloadSupplement?doi=10.1111%2Fnph.16746&file=nph16746-sup-0001-SupInfo.pdf, Hydraulic failure of the plant vascular system is a principal cause of forest die-off under drought. Accurate quantification of this process is essential to our understanding of the physiological mechanisms underpinning plant mortality. Imaging techniques increasingly are applied to estimate xylem cavitation resistance. These techniques allow forin situmeasurement of embolism formation in real time, although the benefits and trade-offs associated with different techniques have not been evaluated in detail. Here we compare two imaging methods, microcomputed tomography (microCT) and optical vulnerability (OV), to standard hydraulic methods for measurement of cavitation resistance in seven woody species representing a diversity of major phylogenetic and xylem anatomical groups. Across the seven species, there was strong agreement between cavitation resistance values (P-50) estimated from visualization techniques (microCT and OV) and between visual techniques and hydraulic techniques. The results indicate that visual techniques provide accurate estimates of cavitation resistance and the degree to which xylem hydraulic function is impacted by embolism. Results are discussed in the context of trade-offs associated with each technique and possible causes of discrepancy between estimates of cavitation resistance provided by visual and hydraulic techniques., This work was supported by an ARC Discovery Projec t(DP170100761) to BC and TJB and an ARC Future Fellowship (FT130101115) to BC. We thank Daniel Hausermann, ChrisHall and Anton Maksimenko from the Australian SynchrotronImaging and Medical Beamline in Melbourne for assisting withthe micro-computed tomography methodology, as well as thetechnical staff at SOLEIL. Travel funding for AG to attendbeamtime at SOLEIL was provided by the International Syn-chrotron Access Program (ISAP) managed by the Australian Syn-chrotron. CMR-D was supported by an Individual Fellowshipfrom the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie grant agree-ment no. 751918-AgroPHYS