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玉米表达的转基因内切-1,4-β-D-葡聚糖酶在农业土壤中的活性及其生态影响

Activity and ecological implications of maize-expressed transgenic endo-1,4-β-D-glucanase in agricultural soils.

作者信息

Kenny Adam J, Wolt Jeffrey D

机构信息

Department of Agronomy, Iowa State University, Ames, Iowa, United States.

出版信息

Environ Toxicol Chem. 2014 Sep;33(9):1996-2003. doi: 10.1002/etc.2645. Epub 2014 Jul 18.

Abstract

Plant expression of thermostable endoglucanase (E1) has been proposed for improved conversion of lignocellulose to ethanol for fuel production. Residues of E1-expressing maize may affect ecological services (e.g., C mineralization and biogeochemical cycling) on soils where they occur. Therefore, the activity of residual E1 was investigated using soils amended with bacterial and plant-solubilized E1 compared with soil endogenous activity and residual activity from a mesostable cellulase (Aspergillus and Trichoderma spp.). An optimized analytical method involving a carboxymethyl cellulose substrate and dinitrosalicylic acid detection effectively assayed endoglucanase activity in amended and unamended soils and was used for determining E1 activity in 3 representative soils. The effect of E1 on soil carbon mineralization was determined by comparing CO(2) evolution from soils amended with transgenic E1-expressing and wild-type maize tissue. Extraction and recovery of the mesostable comparator, bacterial E1, and plant-soluble E1 showed nearly complete loss of exogenous endoglucanase activity within a 24-h period. Carbon mineralization indicated no significant difference between soils amended with either the transgenic E1 or wild-type maize tissue. These results indicate that maize residues expressing up to 30 µg E1/g tissue negligibly affect soil endoglucanase activity and CO(2) respiration for representative soils where transgenic E1 maize may be grown.

摘要

有人提出通过植物表达耐热内切葡聚糖酶(E1)来提高木质纤维素转化为乙醇用于燃料生产的效率。表达E1的玉米残留物可能会影响其生长土壤的生态服务功能(如碳矿化和生物地球化学循环)。因此,与土壤内源活性以及中稳定性纤维素酶(曲霉属和木霉属)的残留活性相比,研究了用细菌和植物溶解的E1改良土壤中残留E1的活性。一种涉及羧甲基纤维素底物和二硝基水杨酸检测的优化分析方法有效地测定了改良和未改良土壤中的内切葡聚糖酶活性,并用于测定3种代表性土壤中的E1活性。通过比较用表达转基因E1的玉米组织和野生型玉米组织改良的土壤中CO₂ 的释放量,确定了E1对土壤碳矿化的影响。中稳定性对照物、细菌E1和植物可溶性E1的提取和回收率表明,外源内切葡聚糖酶活性在24小时内几乎完全丧失。碳矿化表明,用转基因E1或野生型玉米组织改良的土壤之间没有显著差异。这些结果表明,对于可能种植转基因E1玉米的代表性土壤,每克组织表达高达30 µg E1的玉米残留物对土壤内切葡聚糖酶活性和CO₂ 呼吸的影响可忽略不计。

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