Effect of the down-regulation of the high Grain Protein Content (GPC) genes on the wheat transcriptome during monocarpic senescence

<p>Abstract</p> <p>Background</p> <p>Increasing the nutrient concentration of wheat grains is important to ameliorate nutritional deficiencies in many parts of the world. Proteins and nutrients in the wheat grain are largely derived from the remobilization of degraded leaf molecules during monocarpic senescence. The down-regulation of the NAC transcription factor <it>Grain Protein Content </it>(<it>GPC</it>) in transgenic wheat plants delays senescence (>3 weeks) and reduces the concentration of protein, Zn and Fe in the grain (>30%), linking senescence and nutrient remobilization.</p> <p>Based on the early and rapid up-regulation of <it>GPC </it>in wheat flag leaves after anthesis, we hypothesized that this transcription factor is an early regulator of monocarpic senescence. To test this hypothesis, we used high-throughput mRNA-seq technologies to characterize the effect of the <it>GPC </it>down-regulation on the wheat flag-leaf transcriptome 12 days after anthesis. At this early stage of senescence <it>GPC </it>transcript levels are significantly lower in transgenic GPC-RNAi plants than in the wild type, but there are still no visible phenotypic differences between genotypes.</p> <p>Results</p> <p>We generated 1.4 million 454 reads from early senescing flag leaves (average ~350 nt) and assembled 1.2 million into 30,497 contigs that were used as a reference to map 145 million Illumina reads from three wild type and four GPC-RNAi plants. Following normalization and statistical testing, we identified a set of 691 genes differentially regulated by <it>GPC </it>(431 ≥ 2-fold change). Transcript level ratios between transgenic and wild type plants showed a high correlation (<it>R </it>= 0.83) between qRT-PCR and Illumina results, providing independent validation of the mRNA-seq approach. A set of differentially expressed genes were analyzed across an early senescence time-course.</p> <p>Conclusions</p> <p>Monocarpic senescence is an active process characterized by large-scale changes in gene expression which begins considerably before the appearance of visual symptoms of senescence. The mRNA-seq approach used here was able to detect small differences in transcript levels during the early stages of senescence. This resulted in an extensive list of <it>GPC</it>-regulated genes, which includes transporters, hormone regulated genes, and transcription factors. These <it>GPC</it>-regulated genes, particularly those up-regulated during senescence, provide valuable entry points to dissect the early stages of monocarpic senescence and nutrient remobilization in wheat.</p

Growth, grain yield and nitrogen use efficiency of Mediterranean wheat in soils amended with municipal sewage sludge

The application of sewage sludge (SS) to agricultural land can improve soil fertility and physical properties, and enhance crop production. This field study was conducted for two consecutive growing seasons to investigate the influence of SS application on winter wheat growth, grain yield, N accumulation, translocation and use, and on trace elements concentrations in soil and wheat plants under Mediterranean conditions. Treatments consisted of three rates of SS, i.e. 20, 40, and 60 Mg dry weight ha(-1) year(-1), one rate of inorganic fertilizer (IF, 120 kg N ha(-1) year(-1) plus 80 kg P2O5 ha(-1) year(-1)), and an unamended control. The application of SS resulted in tall plants with high early dry matter and N accumulation similar to or significantly higher than those obtained with IF. The lowest SS application rate resulted in grain yield similar to that obtained with IF. Nitrogen use efficiency (NUE) in SS treatments was mainly determined by uptake efficiency, which decreased with increasing SS application rate. Values of NUE and biomass production efficiency with the lowest SS rate were similar to those obtained with IF. SS application resulted in increased concentrations of total and DTPA-extractable trace elements in the soil after the first year, but concentrations were much lower than the regulation limits. Concentrations of Cu, Mn and Zn in wheat plants did not exceed those obtained with IF. Overall, SS could be considered for use as a fertilizer in wheat production systems in the area, serving also as an alternative method of SS disposal