Effect of the down-regulation of the high Grain Protein Content (GPC) genes on the wheat transcriptome during monocarpic senescence.
Effect of the down-regulation of the high Grain Protein Content (GPC) genes on the wheat transcriptome during monocarpic senescence.
复制标题
DOI：
10.1186/1471-2164-12-492
复制
发表时间：
2011-10-07
期刊：
影响因子：
4.4
通讯作者：
Dubcovsky J
中科院分区：
文献类型：
--
作者：
Cantu D;Pearce SP;Distelfeld A;Christiansen MW;Uauy C;Akhunov E;Fahima T;Dubcovsky J
提高小麦籽粒的营养浓度对于改善世界许多地区的营养缺乏非常重要。小麦籽粒中的蛋白质和营养物质主要来自单果衰老过程中降解的叶片分子的再活化。转基因小麦植株中 NAC 转录因子谷物蛋白质含量 (GPC) 的下调可延迟衰老（>3 周），并降低谷物中蛋白质、锌和铁的浓度（>30%），将衰老与养分再利用联系起来。基于花后小麦旗叶中 GPC 的早期和快速上调，我们假设该转录因子是单果衰老的早期调节因子。为了检验这一假设，我们使用高通量 mRNA-seq 技术来表征花后 12 天 GPC 下调对小麦旗叶转录组的影响。在衰老的早期阶段，转基因 GPC-RNAi 植物中的 GPC 转录物水平显着低于野生型植物，但基因型之间仍然没有明显的表型差异。我们从早期衰老的旗叶（平均约 350 nt）中生成了 140 万个 454 个读数，并将 120 万个读数组装成 30,497 个重叠群，这些重叠群用作对来自三种野生型和四种 GPC-RNAi 植物的 1.45 亿个 Illumina 读数进行图谱的参考。经过标准化和统计测试，我们确定了一组 691 个受 GPC 差异调节的基因（431 个≥ 2 倍变化）。转基因植物和野生型植物之间的转录水平比率显示 qRT-PCR 和 Illumina 结果之间存在高度相关性 (R = 0.83)，为 mRNA-seq 方法提供了独立验证。对早期衰老时间过程中的一组差异表达基因进行了分析。单果衰老是一个活跃的过程，其特征是基因表达的大规模变化，其在衰老的视觉症状出现之前就开始了。这里使用的 mRNA-seq 方法能够检测衰老早期阶段转录水平的微小差异。这导致了 GPC 调节基因的广泛列表，其中包括转运蛋白、激素调节基因和转录因子。这些 GPC 调节的基因，特别是那些在衰老过程中上调的基因，为剖析小麦单果衰老和养分再利用的早期阶段提供了有价值的切入点。
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.
登录
查看更多内容
影响因子：
12.3
作者：
Andersson A;Keskitalo J;Sjödin A;Bhalerao R;Sterky F;Wissel K;Tandre K;Aspeborg H;Moyle R;Ohmiya Y;Bhalerao R;Brunner A;Gustafsson P;Karlsson J;Lundeberg J;Nilsson O;Sandberg G;Strauss S;Sundberg B;Uhlen M;Jansson S;Nilsson P
通讯作者：
Nilsson P
影响因子：
5.4
作者：
Echenique, V;Stamova, B;Dubcovsky, J
通讯作者：
Dubcovsky, J
影响因子：
7.2
作者：
GRBIC, V;BLEECKER, AB
通讯作者：
BLEECKER, AB
影响因子：
14.9
作者：
Barnes M;Freudenberg J;Thompson S;Aronow B;Pavlidis P
通讯作者：
Pavlidis P
DOI：
10.1111/j.2517-6161.1995.tb02031.x
发表时间：
1995-01-01
影响因子：
5.8
作者：
BENJAMINI, Y;HOCHBERG, Y
通讯作者：
HOCHBERG, Y