Straw return improves soil carbon pool and dissolved organic matter (DOM) characteristics in black soil. Optimal straw return rate is the key to promoting straw return practices in farmland in Northeast China. The experiment was conducted at the Science and Technology Park of China Grain Storage and Northern Corporation in NenJiang, Heilongjiang Province, straw return at 0 kg hm−2, 3000 kg hm−2, 4500 kg hm−2, and 9000 kg hm−2. In the seventh year of the experiment, we used three-dimensional excitation-emission matrices combined with Parallel Factor analysis to characterize the fluorescence characteristics of DOM of black soils. The results showed substantial improvement in soil physical characteristics and soil organic matter (SOM) following straw return, SOM content rises in proportion to the amount of straw returned, and a significant positive correlation coefficient between water-holding capacity (WHC) (p < 0.001, r = 0.82) and dissolved organic matter (DOC) (p < 0.01, r = 0.77). Moreover, straw return significantly increased the richness of three fluorescent components, namely fulvic acid (UV and visible fulvic acids), humic-like acid, and protein-like (short and long-wavelength tryptophan). The fluorescence intensities of these components were lower in straw treatments than in no straw return. The fluorescence intensities of fulvic and humic acids showed decreasing and increasing trends, respectively, with increasing straw return amount. The fluorescence spectroscopy data of DOC demonstrated the key role of high straw return amounts in enhancing substantially the metabolic activity of soil microorganisms. Overall, straw-returning practices improve soil fertility and can be beneficial for black soil farmlands, with the optimal return rate observed at 4500 kg hm−2.
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