The vascular system of plants consists of two conducting tissues, xylem and phloem, which differentiate from procambium cells. Xylem serves as a transporting system for water and signaling molecules and is formed by sequential developmental processes, including cell division/expansion, secondary cell wall deposition, vacuole collapse, and programmed cell death (PCD). PCD during xylem differentiation is accomplished by degradation of cytoplasmic constituents, and it is required for the formation of hollow vessels, known as tracheary elements (TEs). Our recent study revealed that the small GTPase RabG3b acts as a regulator of TE differentiation through its autophagic activation. By using an Arabidopsis in vitro cell culture system, we showed that autophagy is activated during TE differentiation. Overexpression of a constitutively active RabG3b (RabG3bCA) significantly enhances both autophagy and TE differentiation, which are consistently suppressed in transgenic plants overexpressing a dominant negative form (RabG3bDN) or RabG3bRNAi (RabG3bRNAi), a brassinosteroidinsensitive mutant bri1-301, and an autophagy mutant atg5-1. Wood (called secondary xylem) is the most abundant biomass produced by land plants including Populus and Eucalyptus, and therefore is considered to be one of the most cost-effective and renewable bioenergy resources. In an attempt to enhance xylem differentiation and thus to improve biomass traits in poplars, we generated transgenic poplars overexpressing the RabG3bCA form. As notable phenotypes, both stem height and diameter were increased and xylem area in vascular bundles was significantly expanded in RabG3bCA transgenic poplars compared to control plants. Taken together, these results demonstrate that RabG3b regulates xylem differentiation in both Arabidopsis and Populus. This study enhances our understanding of biological mechanisms underlying wood formation and serve as a framework to engineer the quality and quantity of wood as useful biomass.

An Arabidopsis small GTPase, RabG3b, was previously characterized as a component of autophagy and as a positive regulator for xylem development in Arabidopsis. In this work, we assessed whether RabG3b modulates xylem-associated traits in poplar in a similar way as in Arabidopsis. We generated transgenic poplars (Populus alba x P. tremula var. glandulosa) overexpressing a constitutively active form of RabG3b (RabG3bCA) and performed arrange of morphological, histochemical, and molecular analyses to examine xylogenesis. RabG3bCA transgenic poplars showed increased stem growth due to enhanced xylem development. Autophagic structures were observed in differentiating xyelm cells undergoing programmed cell death (PCD) in wild type poplar, and were more abundant in RabG3bCA transgenic poplar plants and cultured cells. Xylogenic activation was also accompanied by the expression of secondary wall-, PCD-, and autophagy-related genes. Collectively, our results suggest that Arabidopsis RabG3b functions to regulate xylem growth through the activation of autophagy during wood formation in Populus, as does the same in Arabidopsis.