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Deep sequencing transcriptional fingerprinting of rice kernels for dissecting grain quality traits

机译:水稻籽粒的深层测序转录指纹分析

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Rice represents one the most important foods all over the world. In Europe, Italy is the first rice producer and Italian production is driven by tradition and quality. All main rice grain quality traits, like cooking properties, texture, gelatinization temperature, chalkiness and yield, are related to the content and composition of starch and seed-storage proteins in the endosperm and to grain shape. In addition, a number of nutraceutical compounds and allergens are known to have a significant effect on grain quality determination. To investigate the genetic bases underlying the qualitative differences that characterize traditional Italian rice cultivars, a comparative RNA-Seq-based transcriptomic analysis of developing caryopsis was conducted at 14?days after flowering on six popular Italian varieties (Carnaroli, Arborio, Balilla, Vialone Nano, Gigante Vercelli and Volano) phenotypically differing for qualitative grain-related traits. Co-regulation analyses of differentially expressed genes showing the same expression patterns in the six genotypes highlighted clusters of loci up or down-regulated in specific varieties, with respect to the others. Among them, we detected loci involved in cell wall biosynthesis, protein metabolism and redox homeostasis, classes of genes affecting in chalkiness determination. Moreover, loci encoding for seed-storage proteins, allergens or involved in the biosynthesis of specific nutraceutical compounds were also present and specifically regulated in the different clusters. A wider investigation of all the DEGs detected in pair-wise comparisons revealed transcriptional variation, among the six genotypes, for quality-related loci involved in starch biosynthesis (e.g. GBSSI, starch synthases and AGPase), genes encoding for transcription factors, additional seed storage proteins, allergens or belonging to additional nutraceutical compounds biosynthetic pathways and loci affecting grain size. Putative functional SNPs associated to amylose content in starch, gelatinization temperature and grain size were also identified. The present work represents a more extended phenotypic characterization of a set of rice accessions that present a wider genetic variability than described nowadays in literature. The results provide the first transcriptional picture for several of the grain quality differences observed among the Italian rice varieties analyzed and reveal that each variety is characterized by the over-expression of a peculiar set of loci affecting grain appearance and quality. A list of candidates and SNPs affecting specific grain properties has been identified offering a starting point for further works aimed to characterize genes and molecular markers for breeding programs.
机译:大米是全世界最重要的食物之一。在欧洲,意大利是第一大米生产国,意大利的生产受到传统和品质的驱动。稻米的所有主要谷物品质特征,如蒸煮特性,质地,糊化温度,白垩度和产量,都与胚乳中淀粉和种子存储蛋白的含量和组成以及籽粒的形状有关。另外,已知许多营养保健品化合物和过敏原对谷物质量的确定具有重要影响。为了研究构成意大利传统水稻品种特征性差异的遗传基础,在六个流行的意大利品种(Carnaroli,Arborio,Balilla,Vialone Nano)上开花后14天,对发育中的颖果进行了基于RNA-Seq的转录组分析。 ,吉甘特·韦切利(Gigante Vercelli)和沃拉诺(Volano))在表观上与谷物的相关性状不同。差异表达基因的共调控分析显示了六种基因型中相同的表达模式,突显了特定品种相对于其他基因而言上调或下调的基因簇。其中,我们检测到了与细胞壁生物合成,蛋白质代谢和氧化还原稳态有关的基因座,这些基因类别影响白垩度的测定。此外,还存在编码种子存储蛋白,变应原或参与特定营养食品化合物生物合成的基因座,并且在不同簇中受到特定调控。在成对比较中检测到的所有DEG的更广泛调查显示,在六种基因型中,与淀粉生物合成有关的质量相关位点(例如,GBSSI,淀粉合酶和AGPase),编码转录因子的基因,其他种子贮藏位点的转录变异蛋白质,过敏原或属于其他营养化合物的生物合成途径和影响基因位点的位点。还确定了与淀粉中直链淀粉含量,糊化温度和颗粒大小相关的推定功能性SNP。目前的工作代表了一组水稻品种的更广泛的表型特征,这些品种的遗传变异性比当今文献中描述的更为广泛。结果为分析的意大利大米品种中观察到的几种谷物品质差异提供了第一张转录图片,并揭示了每个品种的特征是过表达影响谷物外观和品质的独特位点。确定了影响特定谷物特性的候选基因和单核苷酸多态性的清单,为旨在表征育种计划的基因和分子标记的进一步工作提供了起点。

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