, 2002; Alix et al., 2007). Arabidopsis has been used to visualize infection biology of P. brassicae (Mithen & Magrath, 1992). The availability of synteny maps between Arabidopsis and Brassica spp. has allowed the identification of resistance loci in Brassica spp. first identified in Arabidopsis (Suwabe et al., 2006). Global analysis of host gene expression at different time points postinfection has been possible
using Arabidopsis genome arrays, and this has allowed the identification of host genes that may be important for infection by Plasmodiophora (Siemens et al., 2006). Genes of interest can then be studied further by transforming into Arabidopsis or by utilizing the bank of insertion lines available in Arabidopsis (Puzio et al., 2000; Siemens et al., 2006). Many of the host plants that Polymyxa spp. infect are not well characterized genetically, have fewer genetic tools available
and they have long generation click here times. Also, the roots of cereals can be difficult to visualize by microscopy as they are thicker in diameter than those of Arabidopsis. This can sometimes make the visual detection of Polymyxa in roots difficult. Therefore, if infection of Arabidopsis by Polymyxa spp. can be demonstrated, this could be a valuable tool in increasing our understanding of GSK2118436 supplier plant–Polymyxa interactions. This study aimed to look at the potential for infection of Arabidopsis by Polymyxa spp. under controlled environment conditions using Polymyxa-infested soils. Arabidopsis thaliana ecotypes Landsberg erecta (Ler-0) and Columbia (Col-0) were used for this study (supplied by A. Cuzick, Rothamsted Research, UK). These ecotypes were chosen because they are genetically distinct and mapping populations are available. Seeds were sown into sterile Levingtons No. 2 compost containing Vitamin B12 sand and stratified for 4 days in the dark at 4 °C. Pots were then removed and placed in a greenhouse under short-day length conditions (8 h day at 20 °C, 16 °C night, light levels 200–300 μmol m−2 s−1). Once the seedlings had produced their first true leaves, they
were transferred to 10 cm pots containing infectious soils diluted 1 : 2, soil to sterile sand and grown as before. Two UK soils were used: one from Wiltshire, which was infested with SBCMV (Lyons et al., 2008), and one from Woburn, where Polymyxa was present, but no associated virus had ever been identified (Ward et al., 2005; R. Lyons, pers. commun.). For each soil, five seedlings of each ecotype were planted. Plants were then allowed to grow for 2 months. Flowering bolts were removed upon development to prolong vegetative growth. Roots were removed from pots and vigorously washed in sterile, distilled water. Portions of root were then mounted in sterile water under a coverslip and examined using an Axiophot (Zeiss) light microscope with bright field illumination.