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RNA interference suppression of genes in glycosyl transferase families 43 and 47 in wheat starchy endosperm causes large decreases in arabinoxylan content

机译:小麦淀粉胚乳中糖基转移酶家族43和47的基因的RNA干扰抑制导致阿拉伯木聚糖含量大幅下降

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摘要

The cell walls of wheat (Triticum aestivum) starchy endosperm are dominated by arabinoxylan (AX), accounting for 65% to 70% of the polysaccharide content. Genes within two glycosyl transferase (GT) families, GT43 (IRREGULAR XYLEM9 [IRX9] and IRX14) and GT47 (IRX10), have previously been shown to be involved in the synthesis of the xylan backbone in Arabidopsis, and close homologs of these have been implicated in the synthesis of xylan in other species. Here, homologs of IRX10 TaGT47_2 and IRX9 TaGT43_2, which are highly expressed in wheat starchy endosperm cells, were suppressed by RNA interference (RNAi) constructs driven by a starchy endosperm-specific promoter. The total amount of AX was decreased by 40% to 50% and the degree of arabinosylation was increased by 25% to 30% in transgenic lines carrying either of the transgenes. The cell walls of starchy endosperm in sections of grain from TaGT43_2 and TaGT47_2 RNAi transgenics showed decreased immunolabeling for xylan and arabinoxylan epitopes and approximately 50% decreased cell wall thickness compared with controls. The proportion of AX that was water soluble was not significantly affected, but average AX polymer chain length was decreased in both TaGT43_2 and TaGT47_2 RNAi transgenics. However, the long AX chains seen in controls were absent in TaGT43_2 RNAi transgenics but still present in TaGT47_2 RNAi transgenics. The results support an emerging picture of IRX9-like and IRX10-like proteins acting as key components in the xylan synthesis machinery in both dicots and grasses. Since AX is the main component of dietary fiber in wheat foods, the TaGT43_2 and TaGT47_2 genes are of major importance to human nutrition.
机译:小麦(Triticum aestivum)淀粉质胚乳的细胞壁以阿拉伯木聚糖(AX)为主,占多糖含量的65%至70%。以前已证明两个糖基转移酶(GT)家族中的基因GT43(不规则XYLEM9 [IRX9]和IRX14)和GT47(IRX10)参与拟南芥中木聚糖主链的合成,并且它们的近端同源物已经存在。涉及其他物种木聚糖的合成。在这里,在小麦淀粉状胚乳细胞中高表达的IRX10 TaGT47_2和IRX9 TaGT43_2的同源物被淀粉状胚乳特异性启动子驱动的RNA干扰(RNAi)构建抑制。在携带任一转基因的转基因品系中,AX的总量减少了40%至50%,阿拉伯糖基化程度增加了25%至30%。 TaGT43_2和TaGT47_2 RNAi转基因谷物的淀粉胚乳细胞壁显示出对木聚糖和阿拉伯木聚糖表位的免疫标记降低,与对照相比,细胞壁厚度降低了约50%。在TaGT43_2和TaGT47_2 RNAi转基因中,水溶性AX的比例没有受到显着影响,但平均AX聚合物链长均降低。但是,在对照中看到的长AX链在TaGT43_2 RNAi转基因中不存在,但仍存在于TaGT47_2 RNAi转基因中。这些结果支持了在双子叶植物和草木中,IRX9样和IRX10样蛋白作为木聚糖合成机器中关键成分的新现象。由于AX是小麦食品中膳食纤维的主要成分,因此TaGT43_2和TaGT47_2基因对于人类营养至关重要。

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