Global ubiquitinome profiling identifies NEDD4 as a regulator of Profilin 1 and actin remodelling in neural crest cells.

The ubiquitin ligase NEDD4 promotes neural crest cell (NCC) survival and stem-cell like properties to regulate craniofacial and peripheral nervous system development. However, how ubiquitination and NEDD4 control NCC development remains unknown. Here we combine quantitative analysis of the proteome, transcriptome and ubiquitinome to identify key developmental signalling pathways that are regulated by NEDD4. We report 276 NEDD4 targets in NCCs and show that loss of NEDD4 leads to a pronounced global reduction in specific ubiquitin lysine linkages. We further show that NEDD4 contributes to the regulation of the NCC actin cytoskeleton by controlling ubiquitination and turnover of Profilin 1 to modulate filamentous actin polymerization. Taken together, our data provide insights into how NEDD4-mediated ubiquitination coordinates key regulatory processes during NCC development.

The WtmsDW Locus on Wheat Chromosome 2B Controls Major Natural Variation for Floret Sterility Responses to Heat Stress at Booting Stage.

Heat stress at booting stage causes significant losses to floret fertility (grain set) and hence yield in wheat (Triticum aestivum L.); however, there is a lack of well-characterized sources of tolerance to this type of stress. Here, we describe the genetic analysis of booting stage heat tolerance in a cross between the Australian cultivars Drysdale (intolerant) and Waagan (tolerant), leading to the definition of a major-effect tolerance locus on the short arm of chromosome 2B, Wheat thermosensitive male sterile Drysdale/Waagan (WtmsDW). WtmsDW offsets between 44 and 65% of the losses in grain set due to heat, suggesting that it offers significant value for marker-assisted tolerance breeding. In lines lacking the WtmsDW tolerance allele, peaks in sensitivity were defined with reference to auricle distance, for various floret positions along the spike. Other (relatively minor) floret fertility response effects, including at the Rht-D1 dwarfing locus, were considered likely escape artifacts, due to their association with height and flowering time effects that might interfere with correct staging of stems for heat treatment. Heat stress increased grain set at distal floret positions in spikelets located at the top of the spike and increased the size of spikelets at the base of the spike, but these effects were offset by greater reductions in grain set at other floret positions. Potentially orthologous loci on chromosomes 1A and 1B were identified for heat response of flowering time. The potential significance of these findings for tolerance breeding and further tolerance screening is discussed.

A QTL on the short arm of wheat (Triticum aestivum L.) chromosome 3B affects the stability of grain weight in plants exposed to a brief heat shock early in grain filling.

BACKGROUND: Molecular markers and knowledge of traits associated with heat tolerance are likely to provide breeders with a more efficient means of selecting wheat varieties able to maintain grain size after heat waves during early grain filling. RESULTS: A population of 144 doubled haploids derived from a cross between the Australian wheat varieties Drysdale and Waagan was mapped using the wheat Illumina iSelect 9,000 feature single nucleotide polymorphism marker array and used to detect quantitative trait loci for heat tolerance of final single grain weight and related traits. Plants were subjected to a 3 d heat treatment (37 °C/27 °C day/night) in a growth chamber at 10 d after anthesis and trait responses calculated by comparison to untreated control plants. A locus for single grain weight stability was detected on the short arm of chromosome 3B in both winter- and autumn-sown experiments, determining up to 2.5 mg difference in heat-induced single grain weight loss. In one of the experiments, a locus with a weaker effect on grain weight stability was detected on chromosome 6B. Among the traits measured, the rate of flag leaf chlorophyll loss over the course of the heat treatment and reduction in shoot weight due to heat were indicators of loci with significant grain weight tolerance effects, with alleles for grain weight stability also conferring stability of chlorophyll ('stay-green') and shoot weight. Chlorophyll loss during the treatment, requiring only two non-destructive readings to be taken, directly before and after a heat event, may prove convenient for identifying heat tolerant germplasm. These results were consistent with grain filling being limited by assimilate supply from the heat-damaged photosynthetic apparatus, or alternatively, accelerated maturation in the grains that was correlated with leaf senescence responses merely due to common genetic control of senescence responses in the two organs. There was no evidence for a role of mobilized stem reserves (water soluble carbohydrates) in determining grain weight responses. CONCLUSIONS: Molecular markers for the 3B or 6B loci, or the facile measurement of chlorophyll loss over the heat treatment, could be used to assist identification of heat tolerant genotypes for breeding.

Truncation of grain filling in wheat (Triticum aestivum) triggered by brief heat stress during early grain filling: association with senescence responses and reductions in stem reserves.

Short heat waves during grain filling can reduce grain size and consequently yield in wheat (Triticum aestivum L.). Grain weight responses to heat represent the net outcome of reduced photosynthesis, increased mobilisation of stem reserves (water-soluble carbohydrates, WSC) and accelerated senescence in the grain. To compare their relative roles in grain weight responses under heat, these characteristics were monitored in nine wheat genotypes subjected to a brief heat stress at early grain filling (37°C maximum for 3 days at 10 days after anthesis). Compared with the five tolerant varieties, the four susceptible varieties showed greater heat-triggered reductions in final grain weight, grain filling duration, flag leaf chla and chlb content, stem WSC and PSII functionality (Fv/Fm). Despite the potential for reductions in sugar supply to the developing grains, there was little effect of heat on grain filling rate, suggesting that grain size effects of heat may have instead been driven by premature senescence in the grain. Extreme senescence responses potentially masked stem WSC contributions to grain weight stability. Based on these findings, limiting heat-triggered senescence in the grain may provide an appropriate focus for improving heat tolerance in wheat.