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

ABSTRACT: 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 Grain Protein Content (GPC) 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.Based on the early and rapid up-regulation of GPC 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 GPC down-regulation on the wheat flag-leaf transcriptome 12 days after anthesis. At this early stage of senescence GPC 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.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 GPC (431 ? 2-fold change). Transcript level ratios between transgenic and wild type plants showed a high correlation (R = 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.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 GPC-regulated genes, which includes transporters, hormone regulated genes, and transcription factors. These GPC-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.

SUBMITTER: Cantu D

PROVIDER: S-EPMC3209470 | biostudies-literature | 2011 Oct

REPOSITORIES: biostudies-literature

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

Cantu Dario D Pearce Stephen P SP Distelfeld Assaf A Christiansen Michael W MW Uauy Cristobal C Akhunov Eduard E Fahima Tzion T Dubcovsky Jorge J

BMC genomics 20111007

<h4>Background</h4>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 Grain Protein Content (GPC) in transgenic wheat plants delays senescence (>3 weeks) and reduces the concentration of protein, Zn and Fe in the grain (>3 ...[more]

PMID: 21981858

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Project description:The high performance and stability of wheat genotypes for yield, grain protein content (GPC), and other desirable traits are critical for varietal development and food and nutritional security. Likewise, the genotype by environment (G × E) interaction (GEI) should be thoroughly investigated and favorably utilized whenever genotype selection decisions are made. The present study was planned with the following two major objectives: 1) determination of GEI for some advanced wheat genotypes across four locations (Ludhiana, Ballowal, Patiala, and Bathinda) of Punjab, India; and 2) selection of the best genotypes with high GPC and yield in various environments. Different univariate [Eberhart and Ruessll's models; Perkins and Jinks' models; Wrike's Ecovalence; and Francis and Kannenberg's models], multivariate (AMMI and GGE biplot), and correlation analyses were used to interpret the data from the multi-environmental trial (MET). Consequently, both the univariate and multivariate analyses provided almost similar results regarding the top-performing and stable genotypes. The analysis of variance revealed that variation due to environment, genotype, and GEI was highly significant at the 0.01 and 0.001 levels of significance for all studied traits. The days to flowering, plant height, spikelets per spike, grain per spike, days to maturity, and 1000-grain weight were specifically affected by the environment, whereas yield was mainly affected by the environment and GEI. Genotypes, on the other hand, had a greater impact on the GPC than environmental conditions. As a result, a multi-environmental investigation was necessary to identify the GEI for wheat genotype selection because the GEI was very significant for all of the evaluated traits. Yield, 1000-grain weight, spikelet per spike, and days to maturity were observed to have positive correlations, implying the feasibility of their simultaneous selection for yield enhancement. However, GPC was observed to have a negative correlation with yield. Patiala was found to be the most discriminating environment for both yield and GPC and also the most effective representative environment for GPC, whereas Ludhiana was found to be the most effective representative environment for yield. Eventually, two NILs (BWL7508, and BWL7511) were selected as the top across all environments for both yield and GPC.

Project description:Grain protein content (GPC) is an important quality trait that effectively modulates end-use quality and nutritional characteristics of wheat flour-based food products. The Gpc-B1 gene is responsible for the higher protein content in wheat grain. In addition to higher GPC, the Gpc-B1 is also generally associated with reduced grain filling period which eventually causes the yield penalty in wheat. The main aim of the present study was to evaluate the effect of foliar application of potassium nitrate (PN) and salicylic acid (SA) on the physiological characteristics of a set of twelve genotypes, including nine isogenic wheat lines carrying the Gpc-B1 gene and three elite wheat varieties with no Gpc-B1 gene, grown at wheat experimental area of the Department of Plant Breeding and Genetics, PAU, Punjab, India. The PN application significantly increased the number of grains per spike (GPS) by 6.42 grains, number of days to maturity (DTM) by 1.03 days, 1000-grain weight (TGW) by 1.97 g and yield per plot (YPP) by 0.2 kg/plot. As a result of PN spray, the flag leaf chlorophyll content was significantly enhanced by 2.35 CCI at anthesis stage and by 1.96 CCI at 10 days after anthesis in all the tested genotypes. Furthermore, the PN application also significantly increased the flag leaf nitrogen content by an average of 0.52% at booting stage and by 0.35% at both anthesis and 10 days after anthesis in all the evaluated genotypes. In addition, the yellow peduncle colour at 30 days after anthesis was also increased by 19.08% while the straw nitrogen content was improved by 0.17% in all the genotypes. The preliminary experiment conducted using SA demonstrated a significant increase in DTM and other yield component traits. The DTM increased by an average of 2.31 days, GPS enhanced by approximately 3.17 grains, TGW improved by 1.13g, and YPP increased by 0.21 kg/plot. The foliar application of PN and SA had no significant effect on GPC itself. The findings of the present study suggests that applications of PN and SA can effectively mitigate the yield penalty associated with Gpc-B1 gene by extending grain filling period in the wheat.

Dataset Information

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

Publications

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

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