The cold-induced factor CBF3 mediates root stem cell activity,regeneration,and developmental responses to cold

Plant growth and development involve the specification and regeneration of stem cell niches(SCNs).Although plants are exposed to disparate environmental conditions,how environmental cues affect developmental programs and stem cells is not well understood.Root stem cells are accommodated in meristems in SCNs around the quiescent center(QC),which maintains their activity.Using a combination of genetics and confocal microscopy to trace morphological defects and correlate them with changes in gene expression and protein levels,we show that the cold-induced transcription factor(TF)C-REPEAT BINDING FACTOR 3(CBF3),which has previously been associated with cold acclimation,regulates root development,stem cell activity,and regeneration.CBF3 is integrated into the SHORT-ROOT(SHR)regulatory network,forming a feedback loop that maintains SHR expression.CBF3 is primarily expressed in the root endodermis,whereas the CBF3 protein is localized to other meristematic tissues,including root SCNs.Complementation of cbf3-1 using a wild-type CBF3 gene and a CBF3 fusion with reduced mobility show that CBF3 movement capacity is required for SCN patterning and regulates root growth.Notably,cold induces CBF3,affecting QC activity.Furthermore,exposure to moderate cold around 10℃–12℃promotes root regeneration and QC respecification in a CBF3-dependent manner during the recuperation period.By contrast,CBF3 does not appear to regulate stem cell survival,which has been associated with recuperation from more acute cold(-4℃).We propose a role for CBF3 in mediating the molecular interrelationships among the cold response,stem cell activity,and development.

Reconstruction of lateral root formation through single-cell RNA sequencing reveals order of tissue in itiation

Postembryonic organogenesis is critical for plant development.Underground,lateral roots(LRs)form the bulk of mature root systems,yet the ontogeny of the LR primordium(LRP)is not clear.In this study,we performed the single-cell RNA sequencing through the first four stages of LR formation in Arabidopsis.Our analysis led to a model in which a single group of precursor cells,with a cell identity different from their pericycle origins,rapidly reprograms and splits into a mixed ground tissue/stem cell niche fate and a vascular precursor fate.The ground tissue and stem cell niche fates soon separate and a subset of more specialized vascular cells form sucrose transporting phloem cells that appear to connect to the primary root.We did not detect cells resembling epidermis or root cap,suggesting that outer tissues may form later,preceding LR emergence.At this stage,some remaining initial precursor cells form the primordium flanks,while the rest create a reservoir of pluripotent cells that are able to replace the LR if damaged.Laser ablation of the central and lateral LRP regions showed that remaining cells restart the sequence of tissue initiation to form a LR.Collectively,our study reveals an ontological hierarchy for LR formation with an early and sequential split of main root tissues and stem cells.