原位酶谱可方便、直观地展示土壤中酶活性的二维空间变异,近些年来通过与其他成像技术和分析方法相结合,土壤原位酶谱已发展成为分析土壤微观生态过程及土壤-根系-土壤动物相互作用界面过程的有效方法。然而,土壤原位酶谱技术尚未成熟,亟待深入分析其技术不足、完善技术方法,梳理其在不同领域中的应用现状,拓展并深化其在更多领域研究中的应用前景。基于此,综述并分析了该技术在土壤酶相关研究领域的应用现状,结合原位酶谱技术在底物载体选择与酶活定量方法的优化历程,分析了其存在的问题,发现:①可供研究的酶种类较少;②结果准确性仍有质疑;③在除根际研究外其他领域的研究较少、缺乏系统性,难以直接借鉴。这三者仍是阻碍原位酶谱技术在多领域中推广的最大障碍。综上所述,土壤原位酶谱技术虽仍存在缺陷,但其具有操作简便、测量结果的时空分辨率高等优势,在未来土壤酶学领域研究中仍潜力巨大,研究旨在为推进原位酶谱技术在土壤学和生态学等学科研究中的应用和发展提供理论指导。;In situ soil zymography can conveniently and intuitively display the two-dimensionally spatial variation of enzyme activity. Coupling with other imaging techniques and analysis methods, in situ soil zymography has been developed into a highly targeted method for analyzing soil micro-ecological processes and interface processes of soil-root-soil animal interaction. However, in situ soil zymography is immature. It is urgent to analyze its technical deficiencies and improve the technical methods. It is also necessary to hackle its application in different disciplines nowadays, and furtherly expand and deepen its application prospects. This paper summarizes and analyzes the application status of in situ soil zymography technology in the field of soil enzyme related research, combining with the optimization of substrate carrier selection and enzyme activity quantification of in situ soil zymography technology, analyzing its existent problems. The results show that:① the types of enzymes available for research are less; ② the accuracy of the results is still questionable; ③ the research in other fields except rhizosphere research is less and lack of systematism, so it is difficult to use for reference directly. These are still the biggest obstacles to the application of in situ zymography in many fields. In conclusion, although there are still some defects in in-situ soil zymography, it has great advantages of simple operation and high spatial and temporal resolution of measurement results. This paper provides theoretical guidance for promoting the application and development of soil in situ zymography in soil science and ecology.