Small heat shock proteins (sHSP) berperan penting dalam mempertahankan stabilitas protein dan melindungi sel dari cekaman lingkungan. Protein-protein ini tersebar luas pada tumbuhan dan berkontribusi terhadap toleransi terhadap kondisi yang merugikan, seperti kekeringan, suhu ekstrem, dan cekaman oksidatif. Namun, informasi mengenai famili gen sHSP pada Acorus americanus masih terbatas. Penelitian ini bertujuan untuk mengidentifikasi dan mengkarakterisasi famili gen sHSP pada A. americanus melalui pendekatan in silico. Karakterisasi gen dan protein, analisis motif konservatif, hubungan filogenetik, serta analisis promotor dilakukan untuk mengkaji karakteristik struktural dan regulasi famili gen tersebut. Hasil penelitian menunjukkan bahwa seluruh protein yang teridentifikasi memiliki domain alfa-kristalin (alpha-crystallin domain) yang sangat konservatif, yang mengindikasikan potensi fungsinya sebagai molecular chaperone. Analisis promotor mengidentifikasi berbagai elemen cis-regulator yang berkaitan dengan respon terhadap cahaya, pensinyalan hormon, dan respon terhadap cekaman abiotik. Elemen yang responsif terhadap cahaya merupakan elemen yang paling melimpah, diikuti oleh elemen yang berkaitan dengan asam absisat, metil jasmonat, asam salisilat, kekeringan, dan respons terhadap suhu rendah. Temuan ini menunjukkan bahwa ekspresi gen small heat shock protein pada A. americanus diatur oleh berbagai sinyal lingkungan dan hormonal, yang mendukung potensi perannya dalam adaptasi tumbuhan terhadap kondisi cekaman. Penelitian ini dapat menjadi dasar bagi penelitian lebih lanjut mengenai fungsi gen yang responsif terhadap cekaman pada A. americanus. Konfirmasi lebih lanjut melalui analisis ekspresi dan validasi eksperimental masih diperlukan untuk mengetahui fungsi spesifik masing-masing anggota AaHSP20. Abstract Small heat shock proteins (sHSP) play a crucial role in maintaining protein stability and protecting cells from environmental stress. These proteins are widespread in plants and contribute to tolerance of adverse conditions, such as drought, extreme temperatures, and oxidative stress. However, information regarding the sHSP gene family in Acorus americanus remains limited. This study aims to identify and characterize the sHSP gene family in A. americanus using an in silico approach. Gene and protein characterization, analysis of conserved motifs, phylogenetic relationships, and promoter analysis were conducted to examine the structural and regulatory characteristics of this gene family. The results showed that all identified proteins possess a highly conserved alpha-crystallin domain, indicating their potential function as molecular chaperones. Promoter analysis identified various cis-regulatory elements associated with light response, hormone signaling, and abiotic stress responses. Light-responsive elements were the most abundant, followed by elements associated with abscisic acid, methyl jasmonate, salicylic acid, drought, and low-temperature responses. These findings indicate that the expression of the small heat shock protein gene in A. americanus is regulated by various environmental and hormonal signals, supporting its potential role in plant adaptation to stress conditions. This study can serve as a foundation for further research on the functions of stress-responsive genes in A. americanus. Further confirmation through expression analysis and experimental validation is still needed to determine the specific functions of each member of the AaHSP20 family.