Microdissection to isolate vascular cambium cells in poplar

Abstract

Vascular cambium is the lateral meristem producing xylem cells inwards and phloem cells outwards in plant stem. Thus, in trees, the quality and quantity of wood is a result of highly regulated developmental process depending initially on the vascular cambium cell production. The availability of accurate transcriptomics technologies based on high coverage sequencing raises the level of expectations on tissue sampling to a very high degree. What is the benefit of top-level transcriptomics in wood formation studies if we are using these technologies on raw tissues, mixing cells at the organ level or even higher scale? The presented work describes a nine-step procedure, from standing tree to isolated ray and fusiform cells from cryolyophilized tangential sections of the poplar cambial zone. The aim of this paper is to present a step by step procedure including advices on how to select the optimal tree, how to fell the tree while securing its physiological parameters, how to cryolyophilize and microdissect under binocular, presenting the time schedule of the whole process and RNA analysis.