Analysis of tomato gene promoters activated in syncytia induced in tomato and potato hairy roots by Globodera rostochiensis

Abstract

The potato cyst nematode (Globodera rostochiensis) induces feeding sites (syncytia) in tomato and potato roots. In a previous study, 135 tomato genes up-regulated during G. rostochiensis migration and syncytium development were identified. Five genes (CYP97A29, DFR, FLS, NIK and PMEI) were chosen for further study to examine their roles in plant–nematode interactions. The promoters of these genes were isolated and potential cis regulatory elements in their sequences were characterized using bioinformatics tools. Promoter fusions with the β-glucuronidase gene were constructed and introduced into tomato and potato genomes via transformation with Agrobacterium rhizogenes to produce hairy roots. The analysed promoters displayed different activity patterns in nematode-infected and uninfected transgenic hairy roots.Electronic supplementary materialThe online version of this article (doi:10.1007/s11248-012-9665-4) contains supplementary material, which is available to authorized users.

Hairy Root Transformation Using Agrobacterium rhizogenes as a Tool for Exploring Cell Type-Specific Gene Expression and Function Using Tomato as a Model

“…A. rhizogenes can be used as a tool to determine spatial and temporal aspects of gene expression in plants in order to infer signaling pathways associated with pathogen response. A set of candidate genes were tested in tomato and potato (Solanum tuberosum) for their role in the production of a nematode feeding structure upon infection by the cyst nematode Globodera rostochiensis (Wi sniewska et al, 2013). Additional tools to enable the efficiency of hairy root transformation include the pHairy plasmid, which uses the red fluorescent protein DsRed2 under the control of a cauliflower mosaic virus 35S promoter for visual selection of transgenic hairy roots in soybean (Glycine max).…”

Draft Genome Sequence of Rhizobium rhizogenes Strain ATCC 15834

“…The bacteria was streaked on solid MG/L medium and grown for 2 days at 28°C (7). Cultures of the strain were grown on liquid MG/L for preparation of electrocompetent cells (6) and DNA extraction.…”

“…The Ri T-DNA expression in plants leads to increased biosynthesis of the plant hormone auxin ( 3 ), which affects root growth and promotes lateral root initiation ( 4 ). This ability to induce transgenic infinitely growing root systems has been applied for biotechnology and research uses, including production of phytochemicals and recombinant proteins ( 5 ) and studies of root development ( 6 – 8 ).…”

“…Rhizobium rhizogenes strain ATCC 15834 was obtained from ATCC for hairy root culture work with tomato ( 6 ). The bacteria was streaked on solid MG/L medium and grown for 2 days at 28°C ( 7 ). Cultures of the strain were grown on liquid MG/L for preparation of electrocompetent cells ( 6 ) and DNA extraction.…”

In vitro co-culture of Solanum tuberosum hairy roots with Meloidogyne chitwoodi: structure, growth and production of volatiles

“…Although potato HR have been previously established and used for metabolomic (Valancin et al 2013) and genetic transformation evaluation (Nagy et al 2005), and studies have addressed nematode effect on potato hairy root cultures (Hansen et al 1996;Wiśniewska et al 2013), no previous study established a S. tuberosum HR/M. chitwoodi co-culture system, which requires regular subculturing of the host/pathogen system.…”