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

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

机构信息

Plant Biology and Crop Science, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, United Kingdom.

出版信息

Plant Physiol. 2013 Sep;163(1):95-107. doi: 10.1104/pp.113.222653. Epub 2013 Jul 22.

Abstract

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.

摘要

小麦(Triticum aestivum)淀粉胚乳的细胞壁主要由阿拉伯木聚糖(AX)组成,占多糖含量的 65%至 70%。两个糖基转移酶(GT)家族内的基因,GT43(不规则木质部 9[IRX9]和 IRX14)和 GT47(IRX10),先前已被证明参与拟南芥木聚糖骨架的合成,并且这些基因的密切同源物已被牵连在其他物种的木聚糖合成中。在这里,高度表达于小麦淀粉胚乳细胞中的 IRX10 TaGT47_2 和 IRX9 TaGT43_2 的同源物,被由淀粉胚乳特异性启动子驱动的 RNA 干扰(RNAi)构建体所抑制。携带其中任何一个转基因的转基因系的 AX 总量减少了 40%至 50%,阿拉伯糖基化程度增加了 25%至 30%。在 TaGT43_2 和 TaGT47_2 RNAi 转基因谷物切片的淀粉胚乳细胞壁中,木聚糖和阿拉伯木聚糖表位的免疫标记减少,细胞壁厚度比对照减少了约 50%。AX 的水溶性比例没有明显受到影响,但 TaGT43_2 和 TaGT47_2 RNAi 转基因系的 AX 聚合物链平均长度均降低。然而,在对照中看到的长 AX 链在 TaGT43_2 RNAi 转基因系中不存在,但仍存在于 TaGT47_2 RNAi 转基因系中。结果支持了 IRX9 样和 IRX10 样蛋白作为双子叶植物和禾本科植物木聚糖合成机制关键成分的新兴观点。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 基因对人类营养具有重要意义。

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