Purification and biochemical characterization of recombinant rice UDP-arabinopyranose mutase generated in insect cells.

Plants utilize UDP-arabinofuranose (UDP-Araf) in the biosynthesis of Araf-containing complex carbohydrates. UDP-Araf is synthesized from UDP-arabinopyranose by UDP-arabinopyranose mutases (UAMs). Here we describe the heterologous expression of rice (Oryza sativa) UAM genes in insect cells and report some of their enzymatic properties. Recombinant UAMs might serve as useful tools for the biosynthesis of UDP-Araf and might be better than chemical synthesis.

Efficient construction of cDNA microarrays utilizing normalized cDNA libraries of Arabidopsis thaliana.

We have demonstrated the utility of normalization for efficient cDNA microarray preparation using Arabidopsis as a model. Nonredundant cDNAs including 5722 species were efficiently collected from random 7914 expressed sequence tags (ESTs) in four normalized cDNA libraries. The prepared microarrays were successfully used to monitor gene expression. These methodologies should be applicable to the study of other species in plant biotechnology.

Systematic transient assays of promoter activities for leaf-specific genes identified by gene-expression profiling with cDNA microarrays in Arabidopsis thaliana.

We identified 85 genes highly expressed in leaves using an Arabidopsis cDNA microarray. A vector, pRAB5, was designed to allow cloning and assaying of the promoters. Fifty-one promoters from the selected genes were cloned and then assayed using a microprojectile bombardment and dual luciferase reporter assay system. This system allowed efficient systematic assays of promoter activity.

Active gene expression of a xyloglucan endotransglucosylase/hydrolase gene, XTH9, in inflorescence apices is related to cell elongation in Arabidopsis thaliana.

Regulation of cell wall structure plays a central role in growth and differentiation in plants. Xyloglucan endotransglucosylase/hydrolases (XTHs) that catalyze the cleavage and molecular grafting of xyloglucan chains function in loosening and rearrangement of the cell wall. We have characterized XTH9, a member of the XTH family that was isolated by systematic differential screening for highly expressed genes in shoot apices in Arabidopsis. In the vegetative phase, XTH9 transcripts accumulate in the shoot apex region. In the reproductive phase, transcript levels in shoot apices increase further, and they also are detected in flower buds, flower stalks and internodes bearing flowers. XTH9 expression levels were reduced markedly in mutants such as acl which are characterized by short internodal cell lengths, but recover at permissive temperatures in the temperature-sensitive acl mutants. Differential expression of XTH9 along the inflorescence stem was also detected in pin1 where no lateral organs are formed. These observations suggest that XTH9 expression is coordinated with plant development, including the differentiation from vegetative and reproductive meristems and cell elongation of the inflorescence stem.