Cloning and characterization of cDNA encoding xyloglucan endotransglucosylase in Pennisetum glaucum L.

Abstract

Biomass production in plant is directly related to the amount of intercepted solar radiation by the canopy and available water to the plant. Growth and development of leaves, especially under drought condition, is therefore major determinant of crop productivity. Xyloglucan endotransglucosylase (XET) plays important role in growth and development of plants. XETs are a family of enzymes that mediate construction and restructuring of xyloglucan cross-links, thereby controlling the mechanical properties of cell wall. We cloned complete cDNA of an XET from pearl millet (Pennisetum glaucum L.) and characterized it using in silico comparative genomics and activity assays. The cloned cDNA was 1266 bp in length, encoding a protein with 291 amino acids having signal peptide targeting it to the cell wall. The protein showed xyloglucan endotransglucosylase activity but no hydrolytic activity, therefore, named as PgXET1 as per the convention. The comparative genomics revealed that the functional sites of the enzyme (XET) were highly conserved. Evolutionary studies using phylogenetic tree indicated its grouping with XETs from maize (with >95% bootstrap support), barley, rice, etc. This is the first report on cloning and characterization of an XET (PgXET1) from pearl millet, an important dual-purpose crop.

Key words: Xyloglucan endotransglucosylase, Pennisetum glaucum, pearl millet, primary cell wall, cell expansion, drought tolerance.