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The SCL9 subfamily is a key member of the GRAS family that regulates plant development and stress responses. Nevertheless, the functional role of these genes in the growth and development of poplar still unclear. Here, we reported the six <i>SCL9</i> genes, which were found to be differentially expressed during poplar adventitious root formation. The full-length sequences of <i>PeSCL9</i> genes of ‘Nanlin895’ poplar (<i>Populus deltoids</i> × <i>Populus euramericana</i>) were cloned by the RACE technique All <i>PeSCL9</i> genes lacked introns. RT-qPCR revealed that <i>PeSCL9</i> genes displayed a dynamic expression pattern in the adventitious root of poplar, according to RT-qPCR data. A series of comprehensive genes characteristics analysis were carried out for six genes by bioinformation. Meanwhile, transient expression analysis of the <i>Populus</i> protoplasts showed that all the PeSCL9 proteins were localized in the nucleus. In addition, the degradome and sRNA of ‘Nanlin895’ poplar in combination were used to predict miRNAs that regulate <i>PeSCL9</i>. It was found that miR396a and miR396c may affect <i>PeSCL9</i> expression via cleavage, which was further verified by a transient expression experiment in <i>Populus</i> protoplasts. Overall, the development of poplar adventitious root and other tissues was closely related to these six <i>SCL9</i> genes, and they serve as a starting point for further research into the mechanisms regulating poplar growth and development.