We tested the amplification efficiency before using housekeeping genes. Referring to the previous study of Arabidopsis housekeeping genes (Dheda, 2004; Czechowski, 2005; Hou, 2018), we selected Atactin1 (F: AGGCACCTCTTAACCCTAAAGC, R: GGACAACGGAATCTCTCAGC), Atactin2 (F: TTGTGCTGGATTCTGGTGATGG, R: CCGCTCTCTGCTGTTGTGGTG) and Atactin8 (F: GAATTACCCGACGGACA, R: ACGGTCTGCAATACCT) were tested for efficiency, and we found that the best result was Atactin2. The best housekeeping gene for O. fragrans is OfACT, which has been reported in a previous study (Zhang, 2015). Dheda, K.; Huggett, J.F. ; Bustin, S.A. Validation of housekeeping genes for normalizing RNA expression in real-time PCR. Biotechniques. 2004, 37: 112-119. https://doi.org/10.2144/04371RR03 Czechowski, T.; Stitt, M.; Altmann, T. Genome-wide identification and testing of superior reference genes for transcript normalization in Arabidopsis. Plant Physiol. 2005, 139: 5-17. https://doi.org/10.1104/pp.105.063743 Hou, H.; Jia, H.; Yan, Q.; Wang, X. Overexpression of a SBP-Box Gene (VpSBP16) from Chinese Wild Vitis Species in Arabidopsis Improves Salinity and Drought Stress Tolerance. Int. J. Mol. Sci. 2018, 19, 940. https://doi.org/10.3390/ijms19040940 Zhang, C.; Fu, J.X. ; Wang, Y.G. ; Bao, Z.Y. ; Zhao, H.B. Identification of Suitable Reference Genes for Gene Expression Normalization in the Quantitative Real-Time PCR Analysis of Sweet Osmanthus (Osmanthus fragrans Lour.). Plos one. 2015, 10,1-17. https://doi.org/10.1371/journal.pone.0136355 Point 3: Line 124: generat instead of generate.
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