A visual leaf zymography technique for the in situ examination of plant enzyme activity under the stress of environmental pollution.
A visual leaf zymography technique for the in situ examination of plant enzyme activity under the stress of environmental pollution.
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DOI：
10.1021/acs.jafc.0c03815
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发表时间：
2020-08
影响因子：
6.1
通讯作者：
Yu-Xi Feng;Xin Chen;Yan-Wen Li;Hai-Ming Zhao;L. Xiang;Hui Li;Q. Cai;Nai-Xian Feng;C. Mo;M. Wong
中科院分区：
文献类型：
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作者：
Yu-Xi Feng;Xin Chen;Yan-Wen Li;Hai-Ming Zhao;L. Xiang;Hui Li;Q. Cai;Nai-Xian Feng;C. Mo;M. Wong
本研究建立了一种高效的荧光猝灭方法，用于原位可视化和模拟污染胁迫下植物叶片中木聚糖酶、β-葡萄糖苷酶和磷酸酶活性的空间分布(即叶片酶谱技术)。在LZT中，叶表面饱和了酶特异性荧光底物的膜被孵化，并记录下在紫外光下在膜上产生的荧光图像。为了保证酶活性的准确估计，提出了一种基于图像的建模方法，通过降低噪声来恢复真实图像的形态特征。LZT能在2 h内同时测定48个样品，重现性好。LZT测定结果与常规生化分析方法相当，且具有成本低、操作简便等优点。本文解释了研究LZT在大规模监测中用于评估生态毒性的实际应用所需的理论基础。
This study established a high-efficiency fluorescence quenching approach for the in situ visualization and modeling of the spatial distribution of xylanase, β-glucosidase, and phosphatase activities in plant leaves under pollution stress (namely, the leaf zymography technique, LZT). In the LZT, a membrane saturated with an enzyme-specific fluorescent substrate on the leaf surface was incubated and the fluorescent image generated on the membrane under ultraviolet light was recorded. An image-based modeling method for restoring the morphological traits of the true image by reducing noise was developed to ensure the accurate estimation of enzyme activities. The LZT could simultaneously measure 48 samples within 2 h, with good reproducibility. The results obtained by the LZT were comparable to those obtained by a conventional biochemical analysis method, and presented advantages cost and convenience. This paper explains the theoretical basis required to investigate the realistic application of LZT for assessing ecotoxicity in large-scale monitoring.