Plant Gene and Trait 2024, Vol.15, No.1, 8-14 http://genbreedpublisher.com/index.php/pgt 13 to search for members of the cucumber SBP family (Chen et al., 2020). The cucumber genome file was obtained from the cucumber genome database (ftp://cucurbitgenomics.org/pub/cucurbit/genome/cucumber/Chinese_long/v2/), while the Arabidopsis SBP protein sequences were downloaded from TAIR (https://www.arabidopsis.org/index.jsp). Subsequently, the cucumber SBP obtained from the previous step were confirmed using BLASTP (e-value, 1e-5) in NCBI (https://www.ncbi.nlm.nih.gov/). The SBP functional domains were analyzed using SMART (http://smart.embl.de/) to confirm that the selected proteins were cucumber SBP proteins. The isoelectric point and molecular weight of the cucumber SBP proteins were analyzed using the ProtParam platform (https://web.expasy.org/compute_pi/). 4.2 Chromosomal localization and phylogenetic analysis TBtools was used to identify the location and distribution of cucumber SBPgenes on chromosomes. Phylogenetic analysis was then performed using the SBP protein sequences of cucumber and Arabidopsis thaliana. TheMEGA X software was used to construct a phylogenetic tree using the neighbor-joining (NJ) method with 1 000 bootstrap replicates. Beautification of the phylogenetic tree was performed using Evolview V3 (https://www.evolgenius.info//evolview/#login). 4.3 Analysis of gene structure and conserved protein motifs TBtools was used to identify the gene structure of cucumber SBP genes, while MEME 5.0.5 (http://meme-suite.org/tools/meme) was used to identify the conserved protein motifs of cucumber SBP proteins. 4.4 Expression profiling analysis To investigate the expression profile of cucumber SBP genes in different organs, transcriptome data of various cucumber tissues were obtained from the NCBI website (Accession number: SRP071224), and the analysis methods were followed as described by Wei et al. (2016). TBtools was used to generate a heatmap of the expression profiles of cucumber SBPgenes. Acknowledgments This study was supported by the Shanghai Science and Technology Innovation Action Plan in the Agriculture Field (20392001300), the Shanghai Natural Science Foundation (20ZR1439600), the Young Talents Project of Shanghai Agricultural and Forestry Vocational College (A2-0273-20-01-16), and the Internal Project of Shanghai Agricultural and Forestry Vocational College (KY2-0000-20-01). Authors’ Contributions GDJ, ZQ, and ZWW are the designers and conductors of this experiments. GDJ, ZQ, and XTB performed the data analysis and wrote the draft of the manuscript. ZP and CWJ participated in experimental design and data analysis. ZWW conceived and supervised the project, guided the experimental design, data analysis, manuscript writing, and revising. All authors read and approved the final manuscript. Conflict of Interest Disclosure The authors affirm that this research was conducted without any commercial or financial relationships that could be construed as a potential conflict of interest. References Cannon S.B., Mitra A., Baumgarten A., Young N.D., and May G., 2004, The roles of segmental and tandem gene duplication in the evolution of large gene families in Arabidopsis thaliana, BMC Plant Biol., 4(1): 10. Cardon G., Höhmann S., Klein J., Nettesheim K., Saedler H., and Huijser P., 1999, Molecular characterisation of the Arabidopsis SBP-box genes, Gene, 237(1): 91-104. https://doi.org/10.1016/S0378-1119(99)00308-X PMid:10524240 Chen C., Chen H., Zhang Y., Thomas H.R., Frank M.H., He Y., and Xia R., 2020, TBtools: an integrative toolkit developed for interactive analyses of big biological data, Molecular Plant, 13(8): 1194-1202. https://doi.org/10.1016/j.molp.2020.06.009
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