A High-Throughput Platform for Screening Milligram Quantities of Plant Biomass for Lignocellulose Digestibility
A High-Throughput Platform for Screening Milligram Quantities of Plant Biomass for Lignocellulose Digestibility
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DOI：
10.1007/s12155-009-9074-6
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发表时间：
2010-03-01
影响因子：
3.6
通讯作者：
Walton, Jonathan D.
中科院分区：
文献类型：
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作者：
Santoro, Nicholas;Cantu, Shane L.;Walton, Jonathan D.
发展可行的木质纤维乙醇工业需要在将生物质转化为乙醇的过程中进行多重改进。关键的一步是改进将用作生物质原料的植物。为了方便原料植物的鉴定和评价，有一种方法可以筛选大量单独的植物，以提高对特定预处理和酶的组合的消化率。介绍了一个高通量消化平台(HTDP)，该平台是由美国能源部-五大湖生物能源研究中心(DOEGLBRC)赞助开发的，用于筛选种质资源以提高消化率。该平台的一个关键部件是一个定制的工作站，可以在16小时内研磨和分配超过250种不同的植物组织样品。处理的其他步骤(前处理、酶消化和糖分析)也已经实现了很大程度的自动化，需要36小时。该过程适用于不同的酸性和碱性低温前处理。HTDP的总吞吐量为每周972个独立的生物质样本。该平台的验证是在玉米的棕色中脉突变体上进行的，这些突变体已知具有更高的消化率。进一步的验证是通过筛选大约1,200个T-DNA插入已知或怀疑参与细胞壁生物合成的基因的拟南芥突变系来进行的。有几个品系表现出极显著(p<；0.01)的葡萄糖和木糖释放增加(比平均值高20-40%)。该平台可用于筛选植物群体，以确定木质纤维乙醇应用的优良种质，也可用于筛选突变模式植物群体，以确定影响消化率的特定基因。
The development of a viable lignocellulosic ethanol industry requires multiple improvements in the process of converting biomass to ethanol. A key step is the improvement of the plants that are to be used as biomass feedstocks. To facilitate the identification and evaluation of feedstock plants, it would be useful to have a method to screen large numbers of individual plants for enhanced digestibility in response to combinations of specific pretreatments and enzymes. This paper describes a high-throughput digestibility platform (HTDP) for screening collections of germplasm for improved digestibility, which was developed under the auspices of the Department of Energy-Great Lakes Bioenergy Research Center (DOEGLBRC).A key component of this platform is a custom-designed workstation that can grind and dispense 1-5 mg quantities of more than 250 different plant tissue samples in 16 h. The other steps in the processing ( pretreatment, enzyme digestion, and sugar analysis) have also been largely automated and require 36 h. The process is adaptable to diverse acidic and basic, low-temperature pretreatments. Total throughput of the HTDP is 972 independent biomass samples per week. Validation of the platform was performed on brown midrib mutants of maize, which are known to have enhanced digestibility. Additional validation was performed by screening approximately 1,200 Arabidopsis mutant lines with T-DNA insertions in genes known or suspected to be involved in cell wall biosynthesis. Several lines showed highly significant (p< 0.01) increases in glucose and xylose release (20-40% above the mean). The platform should be useful for screening populations of plants to identify superior germplasm for lignocellulosic ethanol applications and also for screening populations of mutant model plants to identify specific genes affecting digestibility.