Gene amplification, an important mechanism of oncogene activation in breast cancer, can have both prognostic and therapeutic implications. In this work, an attempt is made to identify amplified genes that can be used to improve prognostication in breast cancer. A series of 52 node-negative tumours was screened for genomic gains at 57 loci by array-CGH. A subset of these genes was identified that could divide the series into two divergent outcome groups of either long-term survivors or early disease-related deaths (p = 0.01) using a combination of k-means clustering and statistical analysis. The prognostic significance of amplification of four of the genes (EMS1, TOP2A, CCNE1, and ERBB2) was then evaluated, using fluorescent in situ hybridization on a tissue microarray, in a second larger 'validation' series of 232 tumours with a median follow-up of 4.8 years. Adverse disease-related outcome was associated with amplification of TOP2A (p = 0.004); ERBB2 (p = 0.002); and with the combined amplification of TOP2A, ERBB2, and EMS1 (p = 0.01). EMS1 amplification was more common (26% of cases) than previously reported but, in isolation, had no prognostic significance. Amplification of CCNE1, seen in only 6% of cases, had no prognostic role. These results indicate that the complementary use of array-CGH and tissue microarrays has the potential to help in the identification and validation of molecular markers that can be used to classify breast cancers into different prognostic groups.

The mechanism and expression of resistance to glyphosate at different plant growing temperatures was investigated in an Amaranthus palmeri population (VM1) from a soybean field in Vicuña Mackenna, Cordoba, Argentina. Resistance was not due to reduced glyphosate translocation to the meristem or to EPSPS duplication, as reported for most US samples. In contrast, a proline 106 to serine target-site mutation acting additively with EPSPS over-expression (1.8-fold increase) was respectively a major and minor contributor to glyphosate resistance in VM1. Resistance indices based on LD50 values generated using progenies from a cross between 52 PS106 VM1 individuals were estimated at 7.1 for homozygous SS106 and 4.3 for heterozygous PS106 compared with homozygous wild PP106 plants grown at a medium temperature of 24 °C day/18 °C night. A larger proportion of wild and mutant progenies survived a single commonly employed glyphosate rate when maintained at 30 °C day/26 °C night compared with 20 °C day/16 night in a subsequent experiment. Interestingly, the P106S mutation was not identified in any of the 920 plants analysed from 115 US populations, thereby potentially reflecting the difference in A. palmeri control practices in Argentina and USA.

The implementation of a successful glyphosate resistance management strategy requires a simple and cost-effective method for detecting resistance in key weeds. To date, however, glyphosate resistance is still routinely confirmed via laborious and time consuming whole-plant pot assays using seeds collected at the end of the growing season. Here, we describe a simple, early-season bioassay for detecting evolved glyphosate resistance in grass and broadleaf weeds. It involves transplanting suspected glyphosate resistant seedlings alongside known sensitive and resistant standards into agar containing informative rates of herbicide and recording percentage survival 14 d after plating. The method was validated using sensitive and resistant populations of Lolium, Eleusine, Conyza, and Amaranthus species encompassing the main glyphosate resistance mechanisms, namely, impaired translocation, EPSPS gene duplication, and mutations. The whole plant pot and agar-based seedling tests generated comparable resistance indices in dose-response assays and percentage survival at discriminating glyphosate rates. The method was applied successfully to detect resistance in a rigid ryegrass population collected from a French vineyard well before glyphosate was applied in the field for the current season. Additionally, the test was shown to be highly transferable to several other grass and broadleaf weeds that have evolved resistance to glyphosate. One major attribute of the method is that it is capable of detecting resistance regardless of the mechanism involved. In addition to being very simple, quick and, cost-effective, it allows determination of glyphosate resistance in weeds prior to field application. It thus offers the opportunity for an informed choice of herbicides for effective weed control.

No abstract