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Title:   Author  item
                                            KNIPFER, THORSTEN - University Of Californiaitem
                                            CUNEO, ITALO - University Of Californiaitem
                                            BRODERSEN, CRAIG - Yale Universityitem
                                            McElrone, Andrew
  Submitted to: Plant Physiology
                                    Publication Type: Peer Reviewed Journal
                                    Publication Acceptance Date: 4/19/2016
                                    Publication  Date: 4/22/2016
                                    Citation: Knipfer, T., Cuneo, I., Brodersen, C., Mcelrone, A.J. 2016. In-situ visualization of the dynamics in xylem embolism formation and removal in the absence of root pressure: A study on excised grapevine stems. Plant Physiology. 171(2)1024-1036. doi: 10.1104/pp.16.00136.Interpretive Summary: Gas bubbles form in plant xylem vessels during drought and disrupt long-distance water transport. Some species, like grapevines, have the ability to remove these blockages. Despite evidence suggesting that embolism removal is linked to living cells in the xylem, the underlying mechanism remains controversial and is thought to involve positive pressure generated by roots. Here, we used in situ X-ray micro-tomography (microCT) on excised grapevine stems to show that embolism removal is possible without root pressure, and was qualitatively identical to refilling in intact plants. Overall, our results emphasize that root pressure is not required as a driving force for vessel refilling, and care should be taken when performing measurements on excised plant organs containing living cells as vessel behaviour is not static.
                                Technical Abstract: Gas embolisms formed during drought can disrupt long-distance water transport through plant xylem vessels, but some species have the ability to remove these blockages. Despite evidence suggesting that embolism removal is linked to the presence of vessel-associated parenchyma, the underlying mechanism remains controversial and is thought to involve positive pressure generated by roots. Here, we used in situ X-ray micro-tomography (microCT) on excised grapevine stems to determine if embolism removal is possible without root pressure, and if embolism formation/removal affects vessel functional status following sample excision. Our data show that embolism removal in excised stems was driven by water droplet growth and was qualitatively identical to refilling in intact plants. When stem segments were rehydrated with H2O following excision vessel refilling occurred rapidly (<1 h). The refilling process was substantially slower when polyethylene glycol was added to the H2O source to lower the osmotic potential, thereby providing new support for an osmotically driven refilling mechanism. In contrast, segments not supplied with H2O showed no refilling and increased embolism formation. Dynamic changes in liquid:wall contact angles indicated that the processes of embolism removal (i.e. vessel refilling) by water influx and embolism formation by water efflux were directly linked to the activity of vessel-associated living tissue. Overall, our results emphasize that root pressure is not required as a driving force for vessel refilling, and care should be taken when performing hydraulics measurements on excised plant organs containing living vessel-associated tissue as the vessel behaviour may not be static.


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