Characterization and varied expression of a membrane-bound endo-beta-1,4-glucanase in hybrid poplar.
To understand better the intricacies of secondary cell wall biosynthesis in trees, we investigated changes in cellulose chemistry and ultrastructure manifested by the mis-regulation of the poplar membrane-bound beta-1,4-endoglucanase orthologous to KORRIGAN (AtKOR). We isolated the poplar KORRIGAN gene from hybrid poplar (Populus albaxgrandidentata; designated PaxgKOR) and created a self-complementary (hairpin) RNAi suppression construct using PCR products derived from the gene. Additionally, AtKOR was employed to generate transgenic poplar over-expressing KORRIGAN. It was found that down-regulation leads to moderate to severe defects in plant growth, an irregular xylem (irx) phenotype, and significantly impacts the ultrastructure of the cellulose synthesized. The RNAi-suppressed lines deposited significantly reduced quantities of a more highly crystalline cellulose, while the hemicellulose content and, more specifically, the xylose content increased. In addition, the amount of soluble sucrose in the leaves and xylem decreased. Conversely, the AtKOR transgenics did not significantly alter cell wall development or plant growth parameters, but it did impact the ultrastructure of the cellulose produced, generating trees with less crystalline cellulose and reduced xylose content.
Maloney
Victoria J
VJ
Department of Wood Science, Faculty of Forestry, The University of British Columbia, Vancouver, BC, Canada.
Mansfield
Shawn D
SD
D000595:Amino Acid Sequence; D002241:Carbohydrates / Q000032:analysis; D002473:Cell Wall / Q000378:metabolism; D002480:Cellulase / Q000235:genetics / Q000378:metabolism*; D002482:Cellulose / Q000737:chemistry / Q000648:ultrastructure; D015536:Down-Regulation; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D017705:Lignans / Q000032:analysis; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics; D032107:Populus / Q000201:enzymology* / Q000235:genetics; D034622:RNA Interference; D014197:Trees / Q000254:growth & development*; D052584:Xylem / Q000378:metabolism; D014994:Xylose / Q000737:chemistry
D002241:Carbohydrates; D017705:Lignans; D010940:Plant Proteins; D002482:Cellulose; D014994:Xylose; D002480:Cellulase