Measurement of Inner Bark and Leaf Osmolality.

Sugar transport in the phloem is driven by turgor pressure gradients which are created by osmotic gradients resulting from sugars loaded to the phloem at the source tissue and unloaded at the sink tissue. Therefore, osmolality is a key parameter that can be used to evaluate sugar status and get an indication of the driving force for phloem transport. Here we describe how osmotic concentration measurements from inner bark (practically, the phloem) and needles of trees can be measured. This protocol presents the procedure used by Lintunen et al. (Front Plant Sci 7:726, 2016) and Paljakka et al. (Plant Cell Environ 40:2160-2173, 2017), extended by practical advice and discussion of potential errors and caveats. We describe how to implement this procedure for gymnosperm as well as angiosperm trees. This method uses mechanical sap extraction with a centrifuge from inner bark and leaf samples, which have gone through a deep freeze treatment and thawing. The osmotic potential of these samples is then analyzed with a freezing point or vapor pressure osmometer. The aim of these measurements is to study the spatial and temporal dynamics of phloem function.