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真菌阿魏酸酯酶和β-1,4-内切木聚糖酶在高羊茅中的功能共表达改变了收获后细胞壁的解构。

Functional co-expression of a fungal ferulic acid esterase and a β-1,4 endoxylanase in Festuca arundinacea (tall fescue) modifies post-harvest cell wall deconstruction.

作者信息

Buanafina Marcia M de O, Dalton Sue, Langdon Tim, Timms-Taravella E, Shearer Erica A, Morris Phillip

机构信息

Department of Biology, The Pennsylvania State University, 208 Mueller Laboratory, University Park, PA, 16802, USA,

出版信息

Planta. 2015 Jul;242(1):97-111. doi: 10.1007/s00425-015-2288-2. Epub 2015 Apr 9.

Abstract

Improved post-harvest cell wall deconstruction of tall fescue leaves has been demonstrated by in-planta co-expression of a constitutively expressed ferulic acid esterase together with a senescence-induced β-1,4 endoxylanase. Tall fescue plants (Festuca arundinacea) constitutively expressing vacuole- or apoplast-targeted ferulic acid esterase from Aspergillus niger were retransformed with a senescence-induced and apoplast-targeted β-1,4 endo-xylanase from Trichoderma reesei. Enzyme activities in co-expressing plants stabilized after repeated vegetative propagation, with xylanase activity in senescent leaves increasing and ferulic acid esterase activity decreasing after tillering. Plants co-expressing both enzymes in the apoplast, with the lowest levels of ferulate monomers and dimers and the lowest levels of cell wall arabinoxylans, released ten times more cell wall hydroxycinnamic acids and five times more arabinoxylan from the cell wall on autodigestion compared to expression of ferulic acid esterase or xylanase alone. These plants also showed a 31 % increase in cellulase-mediated release of reducing sugars, a 5 % point increase in in vitro dry matter digestibility and a 23 % increase in acetyl bromide-soluble lignin. However, plant growth was adversely affected by expressing FAE in the apoplast, giving plants with narrower shorted leaves, and a 71 % decrease in biomass.

摘要

通过在植物体内共表达组成型表达的阿魏酸酯酶和衰老诱导的β-1,4内切木聚糖酶,已证明高羊茅叶片收获后的细胞壁解构得到改善。组成型表达来自黑曲霉的液泡靶向或质外体靶向阿魏酸酯酶的高羊茅植物(Festuca arundinacea),用来自里氏木霉的衰老诱导且质外体靶向的β-1,4内切木聚糖酶进行再次转化。共表达植物中的酶活性在反复营养繁殖后稳定下来,分蘖后衰老叶片中的木聚糖酶活性增加,阿魏酸酯酶活性降低。在质外体中共表达这两种酶的植物,其阿魏酸单体和二聚体水平最低,细胞壁阿拉伯木聚糖水平最低,与单独表达阿魏酸酯酶或木聚糖酶相比,自消化时从细胞壁释放的细胞壁羟基肉桂酸多十倍,阿拉伯木聚糖多五倍。这些植物还显示,纤维素酶介导的还原糖释放增加31%,体外干物质消化率提高5个百分点,乙酰溴可溶性木质素增加23%。然而,在质外体中表达FAE对植物生长有不利影响,使植物叶片变窄变短,生物量减少71%。

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