Institute Jean-Pierre Bourgin INRA Centre de Versailles-Grignon, Route de Saint-Cyr 10, Versailles Cedex, 78026, France.
Department Plant Breeding and Genetics, Max Planck Institute for Plant Breeding Research, Carl von-Linné-Weg 10, Köln, 50829, Germany.
Plant Cell Environ. 2018 Aug;41(8):1895-1911. doi: 10.1111/pce.13330. Epub 2018 Jun 19.
Genes controlling differences in seed longevity between 2 barley (Hordeum vulgare) accessions were identified by combining quantitative genetics "omics" technologies in near isogenic lines (NILs). The NILs were derived from crosses between the spring barley landraces L94 from Ethiopia and Cebada Capa from Argentina. A combined transcriptome and proteome analysis on mature, nonaged seeds of the 2 parental lines and the L94 NILs by RNA-sequencing and total seed proteomic profiling identified the UDP-glycosyltransferase MLOC_11661.1 as candidate gene for the quantitative trait loci on 2H, and the NADP-dependent malic enzyme (NADP-ME) MLOC_35785.1 as possible downstream target gene. To validate these candidates, they were expressed in Arabidopsis under the control of constitutive promoters to attempt complementing the T-DNA knockout line nadp-me1. Both the NADP-ME MLOC_35785.1 and the UDP-glycosyltransferase MLOC_11661.1 were able to rescue the nadp-me1 seed longevity phenotype. In the case of the UDP-glycosyltransferase, with high accumulation in NILs, only the coding sequence of Cebada Capa had a rescue effect.
通过在近等基因系 (NILs) 中结合定量遗传学“组学”技术,鉴定了控制 2 个大麦 (Hordeum vulgare) 品系间种子寿命差异的基因。这些 NILs 是由埃塞俄比亚春大麦地方品种 L94 和阿根廷 Cebada Capa 之间的杂交产生的。对 2 个亲本系和 L94 NILs 的成熟、未老化种子进行联合转录组和蛋白质组分析,通过 RNA-seq 和总种子蛋白质组谱分析,确定 UDP-糖基转移酶 MLOC_11661.1 为 2H 上数量性状位点的候选基因,NADP 依赖性苹果酸酶 (NADP-ME) MLOC_35785.1 为可能的下游靶基因。为了验证这些候选基因,它们在拟南芥中受组成型启动子的控制表达,试图互补 nadp-me1 T-DNA 敲除系。NADP-ME MLOC_35785.1 和 UDP-糖基转移酶 MLOC_11661.1 都能够挽救 nadp-me1 种子寿命表型。在 UDP-糖基转移酶的情况下,在 NILs 中积累量较高,只有 Cebada Capa 的编码序列具有挽救作用。