A variable region of the Sugarcane Bacilliform Virus (SCBV) genome can be used to generate promoters for transgene expression in sugarcane

Abstract

Four promoters derived from sugarcane bacilliform virus (SCBV) were compared and characterised. Three were obtained by PCR amplification of purified virion DNA extracted from three sugarcane cultivars. The fourth promoter was obtained by subcloning from an almost genome-length clone of SCBV. All promoters were able to drive stable expression of beta-glucuronidase in sugarcane. The PCR-derived promoter sequences shared more DNA homology with banana streak virus than to the subcloned SCBV. The subcloned promoter was the strongest expressing and was able to drive reporter gene expression in vitro and in the leaves, meristems and roots of glasshouse-grown sugarcane. Expression levels were at least equal to or higher than those measured for the maize polyubiquitin promoter.

Genetic Engineering of Saccharum

“…The number of promoters available for sugarcane transformation is very limited, and most of them are of constitutive nature Wei 2003, 2004 ;Braithwaite et al 2004 ;Christensen et al 1992 ;Wei et al 1999Wei et al , 2003Yang et al 2003 ) . However, when some of these promoters were reintroduced into sugarcane, they were transcriptionally silenced (Wei et al 2003 ) .…”

“…Reporter/scorable markers such as b -glucuronidase (GUS), GFP, yellow fl uorescent protein (EYFP), luciferase (LUC), and maize anthocyanin (ANT) regulatory elements have been used for the development and optimization of transformation to allow visual selection of transient and stable integrations (Arencibia et al 1995 ;Bower and Birch 1992 ;Bower et al 1996 ;Elliott et al 1998 ) , functional analysis (Braithwaite et al 2004 ;Damaj et al 2010 ;Gallo-Meagher and Irvine 1993 ;Liu et al 2003 ;Wei et al 2003 ) , subcellular targeting studies (Gnanasambandam and Birch 2004 ;Gnanasambandam et al 2007 ) , dissection of transgene silencing (Birch et al 2010a ) , and determination of terminator ef fi ciencies of constructs (Beyene et al 2011 ).…”

Sugarcane DIRIGENT and O-METHYLTRANSFERASE promoters confer stem-regulated gene expression in diverse monocots

“…In particular, the few promoters isolated and characterized in sugarcane, a major sucrose accumulator and biomass producer, are of a constitutive nature. These include promoters from ubiquitin 4 and 9 genes (Wei et al 1999Albert and Wei 2003), an elongation factor 1 gene, a proline-rich protein-encoding gene , and Sugarcane bacilliform virus (Tzafrir et al 1998;Braithwaite et al 2004). Tissue-speciWc monocot promoters have been developed that target gene expression in leaf (Nomura et al 2000), root (Yu et al 2007), epidermis (Altpeter et al 2005) and anther tapetum (Luo et al 2006).…”

Significant plant virus diseases in India and a glimpse of modern disease management technology

“…These germplasms were long used for breeding purposes, which disseminated these viruses. Furthermore, during a long coexistence, part of the genome of some viral pathogens became integrated within the host as in sugarcane where propagation is vegetative (Braithwaite et al 2004). In such cases, the results of PCR are deceptive, and ELISA/ISEM or Western blotting tests are more dependable.…”

“…DNA sequencing can reveal the extent of genetic variation between isolates. When reverse transcriptase and ribonuclease H amplicons from SCBV isolated from three noble canes were sequenced (Braithwaite et al 2004;Geijskes et al 2002), variability was revealed not just among the SCBV isolates from different canes, but also among isolates within individual cane plants. Thus, conserved or degenerate primers may not reliably prime the amplification of all the isolates.…”