Buanafina Marcia M de O, Langdon Tim, Hauck Barbara, Dalton Sue, Morris Phillip
Plant, Animal and Microbial Science Department, Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth SY23 3EB, UK.
Plant Biotechnol J. 2008 Apr;6(3):264-80. doi: 10.1111/j.1467-7652.2007.00317.x. Epub 2007 Dec 17.
In the cell walls of forage grasses, ferulic acid is esterified to arabinoxylans and participates with lignin monomers in oxidative coupling pathways to generate ferulate-polysaccharide-lignin complexes that cross-link the cell wall. Such cross-links hinder cell wall degradation by ruminant microbes, reducing plant digestibility. In this study, genetically modified Festuca arundinacea plants were produced expressing an Aspergillus niger ferulic acid esterase (FAEA) targeted to the vacuole. The rice actin promoter proved to be effective for FAEA expression, as did the cauliflower mosaic virus (CaMV) 35S and maize ubiquitin promoters. Higher levels of expression were, however, found with inducible heat-shock and senescence promoters. Following cell death and subsequent incubation, vacuole-targeted FAEA resulted in the release of both monomeric and dimeric ferulic acids from the cell walls, and this was enhanced several fold by the addition of exogenous endo-1,4-beta-xylanase. Most of the FAEA-expressing plants showed increased digestibility and reduced levels of cell wall esterified phenolics relative to non-transformed plants. It is concluded that targeted FAEA expression is an effective strategy for improving wall digestibility in Festuca and, potentially, other grass species used for fodder or cellulosic ethanol production.
在饲草的细胞壁中,阿魏酸被酯化到阿拉伯木聚糖上,并与木质素单体参与氧化偶联途径,以生成将细胞壁交联在一起的阿魏酸 - 多糖 - 木质素复合物。这种交联阻碍反刍动物微生物对细胞壁的降解,降低了植物的消化率。在本研究中,培育出了表达靶向液泡的黑曲霉阿魏酸酯酶(FAEA)的转基因高羊茅植株。水稻肌动蛋白启动子被证明对FAEA的表达有效,花椰菜花叶病毒(CaMV)35S启动子和玉米泛素启动子也是如此。然而,在诱导型热休克和衰老启动子作用下发现了更高水平的表达。在细胞死亡及随后的孵育过程中,靶向液泡的FAEA导致细胞壁中单体和二聚体阿魏酸的释放,并且通过添加外源内切 - 1,4 - β - 木聚糖酶,这种释放增强了几倍。相对于未转化的植株,大多数表达FAEA的植株显示出消化率提高且细胞壁酯化酚类物质水平降低。得出的结论是,靶向FAEA表达是提高高羊茅以及潜在地提高用于饲料或纤维素乙醇生产的其他草种细胞壁消化率的有效策略。