Morin, H. | Search bioRxiv

bioRxiv2024

M2WISH: an easy and efficient protocol for whole-mount mRNA in situ hybridization that allows 3D cell resolution of gene expression in Arabidopsis thaliana.

Gene expression analysis is essential for understanding the mechanisms involved in plant development. Here, we developed M2WISH, a protocol based on MicroWave treatment for Wholemount mRNA In Situ Hybridization in Arabidopsis. By permeabilizing tissues without damaging cellular organisation this protocol results in high and homogeneous hybridization yields that enables systematic analysis of gene expression dynamics. Moreover, when combined with cellular histochemical staining, M2WISH provides a cellular resolution of gene expression on roots, aerial meristems, leaves and embryos in the seed. We applied M2WISH to study the spatial dynamics of WUSCHEL (WUS) and CLAVATA3 (CLV3) expression during in vitro meristematic conversion of roots into shoot apical meristems. Thus, we showed that shoot apical meristems could arise from two different types of root structures that differed by their CLV3 gene expression patterns. We constructed 3D cellular representations of WUS and CLV3 gene co-expression pattern, and stressed the variability inherent to meristem conversion. Thus, this protocol generates a large amount of data on the localization of gene expression, which can be used to model complex systems.

plant biology↗

bioRxiv2020

Driving factors of conifer regeneration dynamics in eastern Canadian boreal old-growth forests

Old-growth forests play a major role in conserving biodiversity, protecting water resources, sequestrating carbon, and these forests are indispensable resources for indigenous societies. To preserve the ecosystem services provided by these boreal ecosystems, it becomes necessary to develop novel silvicultural practices capable of emulating the natural dynamics and structural attributes of old-growth forests. The success of these forest management strategies depends on developing an accurate understanding of natural regeneration dynamics. Our goal was therefore to identify the main patterns and the drivers involved in the regeneration dynamics of old-growth forests, placing our focus on boreal stands dominated by black spruce (Picea mariana (L.) Mill.) and balsam fir (Balsam fir (L.) Mill.) in eastern Canada. We sampled 71 stands in a 2200 km2 study area located within Quebecs boreal region. For each stand, we noted tree regeneration (seedlings and saplings), structural attributes (diameter distribution, deadwood volume, etc.), and abiotic (topography and soil) factors. We observed that secondary disturbance regimes and topographic constraints were the main drivers of balsam fir and black spruce regeneration. Furthermore, the regeneration dynamics of black spruce appeared more complex than those of balsam fir. We observed distinct phases of seedling production first developing within the understory, then seedling growth when gaps opened in the canopy, followed by progressive canopy closure. Seedling density, rather than the sapling density, had a major role in explaining the ability of black spruce to fill the canopy following a secondary disturbance. The density of balsam fir seedlings and saplings was also linked to the abundance of balsam fir trees at the stand level. This research helps explain the complexity of old-growth forest dynamics where many ecological factors interact at multiple temporal and spatial scales. This study also improves our understanding of ecological processes within native old-growth forests and identifies the key factors to consider when ensuring the sustainable management of old-growth boreal stands.

ecology↗