Systematic analysis of specific and nonspecific auxin effects on endocytosis and trafficking

The phytohormone auxin and its directional transport through tissues are intensively studied. However, a mechanistic understanding of auxin-mediated feedback on endocytosis and polar distribution of PIN auxin transporters remains limited due to contradictory observations and interpretations. Here, we used state-of-the-art methods to reexamine the auxin effects on PIN endocytic trafficking. We used high auxin concentrations or longer treatments versus lower concentrations and shorter treatments of natural indole-3-acetic acid (IAA) and synthetic naphthalene acetic acid (NAA) auxins to distinguish between specific and nonspecific effects. Longer treatments of both auxins interfere with Brefeldin A-mediated intracellular PIN2 accumulation and also with general aggregation of endomembrane compartments. NAA treatment decreased the internalization of the endocytic tracer dye, FM4-64; however, NAA treatment also affected the number, distribution, and compartment identity of the early endosome/trans-Golgi network, rendering the FM4-64 endocytic assays at high NAA concentrations unreliable. To circumvent these nonspecific effects of NAA and IAA affecting the endomembrane system, we opted for alternative approaches visualizing the endocytic events directly at the plasma membrane (PM). Using total internal reflection fluorescence microscopy, we saw no significant effects of IAA or NAA treatments on the incidence and dynamics of clathrin foci, implying that these treatments do not affect the overall endocytosis rate. However, both NAA and IAA at low concentrations rapidly and specifically promoted endocytosis of photo-converted PIN2 from the PM. These analyses identify a specific effect of NAA and IAA on PIN2 endocytosis, thus, contributing to its polarity maintenance and furthermore illustrate that high auxin levels have nonspecific effects on trafficking and endomembrane compartments.

Natural and synthetic auxins affect various aspects of the endomembrane system at high concentrations, but promote clathrin-mediated endocytosis of the PIN2 auxin transporter at low concentrations.     

Genetic diversity and investigation of polledness in divergent goat populations using 52 088 SNPs

The recent availability of a genome‐wide SNP array for the goat genome dramatically increases the power to investigate aspects of genetic diversity and to conduct genome‐wide association studies in this important domestic species. We collected and analysed genotypes from 52 088 SNPs in Boer, Cashmere and Rangeland goats that had both polled and horned individuals. Principal components analysis revealed a clear genetic division between animals for each population, and model‐based clustering successfully detected evidence of admixture that matched aspects of their recorded history. For example, shared co‐ancestry was detected, suggesting Boer goats have been introgressed into the Rangeland population. Further, allele frequency data successfully tracked the altered genetic profile that has taken place after 40 years of breeding Australian Cashmere goats using the Rangeland animals as the founding population. Genome‐wide association mapping of the POLL locus revealed a strong signal on goat chromosome 1. The 769‐kb critical interval contained the polled intersex syndrome locus, confirming the genetic basis in non‐European animals is the same as identified previously in Saanen goats. Interestingly, analysis of the haplotypes carried by a small set of sex‐reversed animals, known to be associated with polledness, revealed some animals carried the wild‐type chromosome associated with the presence of horns. This suggests a more complex basis for the relationship between polledness and the intersex condition than initially thought while validating the application of the goat SNP50 BeadChip for fine‐mapping traits in goat.     

Inter‐annual site fidelity and breeding origins of Gray‐cheeked Thrushes in white sand forests of the Peruvian Amazon

White sand terra firme forests are unusual ecosystems scattered across Amazonia, covering just 3% of the basin. These forests differ from surrounding forests in their scleromorphic vegetation, low nutrient content, and propensity to harbor endemics. We report the capture of 62 Gray‐cheeked Thrushes (Catharus minimus) during a study of the understory avifauna of Amazonian white sand forests near Iquitos, Peru, conducted from 20 June to 8 December 2010–2012. We captured and banded Gray‐cheeked Thrushes in white sand (N = 57) and adjacent weathered clay (N = 5) terra firme forests. Sampling for three consecutive days at 19 different sites each year, the inter‐annual site fidelity rate of Gray‐cheeked Thrushes was 4.8% (N = 3). One bird banded in 2010 was recaptured in 2012. Of the 62 birds, 19.3% (N = 12) were recaptured on subsequent days. All recaptures were in white sand forests. The 19.3% recapture rate of Gray‐cheeked Thrushes from sites re‐sampled no more than 2 d in a given year suggests the presence of settled and perhaps territorial birds. Using rectrices from 12 Gray‐cheeked Thrushes, stable‐hydrogen isotope analyses (δ²H) suggest that the geographic breeding or natal origin of all sampled birds was likely northwestern North America. Our results suggest that Gray‐cheeked Thrushes exhibit site fidelity and may concentrate in white sand forests—an uncommon and scattered ecosystem type in western Amazonia. However, annual tracking of individual Gray‐cheeked Thrushes is needed to fully assess regional patterns of settlement and movement, and the connectivity between breeding and wintering areas.