{"title":"二维土壤酶谱:考虑 pH 值的空间变化","authors":"Andrey Guber, Alexandra Kravchenko","doi":"10.1016/j.rhisph.2024.100862","DOIUrl":null,"url":null,"abstract":"<div><p><span>Soil zymography is commonly used to quantify spatial distribution of hydrolytic </span>enzyme activities<span> on soil and plant root surfaces. It is recommended to adjust pH in zymography substrates and calibration solutions with respect to soil/root pH. However, pH values may vary greatly within a few mm of plant rhizosphere, potentially altering the distribution of pH in zymography membranes. Despite the fact that the effect of pH on the calibration of zymography membranes is generally known, its potential impact on zymography results is unaccounted for in processing zymography images and calculations of enzyme activity. In this study we assessed the effect of pH variations on the persistency of the methylumbelliferone (MUF) calibration. The studied pH values ranged from 4.5 to 7.5. The MUF calibration curves greatly deviated from that at a reference pH of 6.5, with a marked nonlinear increase of deviation with greater membrane brightness. We suggest that the problem can be partially alleviated by reducing the membrane incubation time. However, such deviations suggest the need for a more comprehensive resolution via mapping pH and using pH-specific calibrations to process zymography images. We developed a MATLAB code to implement a pixel-based correction of enzyme activity for pH in processing time-lapse zymography images.</span></p></div>","PeriodicalId":48589,"journal":{"name":"Rhizosphere","volume":"29 ","pages":"Article 100862"},"PeriodicalIF":3.4000,"publicationDate":"2024-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"2-D soil zymography: Accounting for the spatial variation of pH\",\"authors\":\"Andrey Guber, Alexandra Kravchenko\",\"doi\":\"10.1016/j.rhisph.2024.100862\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p><span>Soil zymography is commonly used to quantify spatial distribution of hydrolytic </span>enzyme activities<span> on soil and plant root surfaces. It is recommended to adjust pH in zymography substrates and calibration solutions with respect to soil/root pH. However, pH values may vary greatly within a few mm of plant rhizosphere, potentially altering the distribution of pH in zymography membranes. Despite the fact that the effect of pH on the calibration of zymography membranes is generally known, its potential impact on zymography results is unaccounted for in processing zymography images and calculations of enzyme activity. In this study we assessed the effect of pH variations on the persistency of the methylumbelliferone (MUF) calibration. The studied pH values ranged from 4.5 to 7.5. The MUF calibration curves greatly deviated from that at a reference pH of 6.5, with a marked nonlinear increase of deviation with greater membrane brightness. We suggest that the problem can be partially alleviated by reducing the membrane incubation time. However, such deviations suggest the need for a more comprehensive resolution via mapping pH and using pH-specific calibrations to process zymography images. We developed a MATLAB code to implement a pixel-based correction of enzyme activity for pH in processing time-lapse zymography images.</span></p></div>\",\"PeriodicalId\":48589,\"journal\":{\"name\":\"Rhizosphere\",\"volume\":\"29 \",\"pages\":\"Article 100862\"},\"PeriodicalIF\":3.4000,\"publicationDate\":\"2024-02-02\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Rhizosphere\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2452219824000156\",\"RegionNum\":3,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"PLANT SCIENCES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Rhizosphere","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2452219824000156","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"PLANT SCIENCES","Score":null,"Total":0}
2-D soil zymography: Accounting for the spatial variation of pH
Soil zymography is commonly used to quantify spatial distribution of hydrolytic enzyme activities on soil and plant root surfaces. It is recommended to adjust pH in zymography substrates and calibration solutions with respect to soil/root pH. However, pH values may vary greatly within a few mm of plant rhizosphere, potentially altering the distribution of pH in zymography membranes. Despite the fact that the effect of pH on the calibration of zymography membranes is generally known, its potential impact on zymography results is unaccounted for in processing zymography images and calculations of enzyme activity. In this study we assessed the effect of pH variations on the persistency of the methylumbelliferone (MUF) calibration. The studied pH values ranged from 4.5 to 7.5. The MUF calibration curves greatly deviated from that at a reference pH of 6.5, with a marked nonlinear increase of deviation with greater membrane brightness. We suggest that the problem can be partially alleviated by reducing the membrane incubation time. However, such deviations suggest the need for a more comprehensive resolution via mapping pH and using pH-specific calibrations to process zymography images. We developed a MATLAB code to implement a pixel-based correction of enzyme activity for pH in processing time-lapse zymography images.
RhizosphereAgricultural and Biological Sciences-Agronomy and Crop Science
CiteScore
5.70
自引率
8.10%
发文量
155
审稿时长
29 days
期刊介绍:
Rhizosphere aims to advance the frontier of our understanding of plant-soil interactions. Rhizosphere is a multidisciplinary journal that publishes research on the interactions between plant roots, soil organisms, nutrients, and water. Except carbon fixation by photosynthesis, plants obtain all other elements primarily from soil through roots.
We are beginning to understand how communications at the rhizosphere, with soil organisms and other plant species, affect root exudates and nutrient uptake. This rapidly evolving subject utilizes molecular biology and genomic tools, food web or community structure manipulations, high performance liquid chromatography, isotopic analysis, diverse spectroscopic analytics, tomography and other microscopy, complex statistical and modeling tools.