The glutamate receptor gene GLR3.3:A bridge of calciummediated root development in poplar

Poplar is one of the fastest-growing temperate trees in the world and is widely used in ornamental horticulture for shade.The root is essential for tree growth and development and its utilization potential is huge.Calcium(Ca),as a signaling molecule,is involved in the regulation of plant root development.However,the detailed underlying regulatory mechanism is elusive.In this study,we analyzed the morphological and transcriptomic variations of 84K poplar(Populus alba×P.glandulosa)in response to different calcium concentrations and found that low Ca^(2+)(1 mmol·L^(-1))promoted lateral root development,while deficiency(0.1 mmol·L^(-1)Ca^(2+))inhibited lateral root development.Co-expression analysis showed that Ca^(2+)channel glutamate receptors(GLRs)were present in various modules with significance for root development.Two GLR paralogous genes,PagGLR3.3a and Pag GLR3.3b,were mainly expressed in roots and up-regulated under Ca^(2+)deficiency.The CRISPR/Cas9-mediated signal gene(crispr-PagGLR3.3a,PagGLR3.3b)and double gene(crispr-PagGLR3.3ab)mutants presented more and longer lateral roots.Anatomical analysis showed that crispr-PagGLR3.3ab plants had more xylem cells and promoted the development of secondary vascular tissues.Further transcriptomic analysis suggested that knockout of PagGLR3.3a and PagGLR3.3b led to the up-regulation of several genes related to protein phosphorylation,auxin efflux,lignin and hemicellulose biosynthesis as well as transcriptional regulation,which might contribute to lateral root growth.This study not only provides novel insight into how the Ca^(2+)channels mediated root growth and development in trees,but also provides a directive breeding of new poplar species for biofuel and bioenergy production.