The Agronomic Traits,Alkaloids Analysis,FT-IR and 2DCOS-IR Spectroscopy Identification of the Low-Nicotine-Content Nontransgenic Tobacco Edited by CRISPR–Cas9 |
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| Authors: | Jianduo Zhang Qun Zhou Dongheyu Zhang Guangyu Yang Chengming Zhang Yuping Wu Yong Xu Jianhua Chen Weisong Kong Guanghui Kong Jin Wang |
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| Institution: | 1.Yunnan Key Laboratory of Tobacco Chemistry, Research and Development Center, China Tobacco Yunnan Industrial Co., Ltd., Kunming 650231, China; (J.Z.); (G.Y.); (C.Z.); (Y.X.); (J.C.); (W.K.);2.Department of Chemistry, Tsinghua University, Beijing 100084, China;3.Department of Electrical Engineering, Tsinghua University, Beijing 100084, China;4.Yunnan Academy of Tobacco Agricultural Science, Kunming 650031, China; (Y.W.); (G.K.) |
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| Abstract: | In this study, the agricultural traits, alkaloids content and Fourier transform infrared spectroscopy (FT-IR) and two-dimensional correlation infrared spectroscopy (2DCOS-IR) analysis of the tobacco after Berberine Bridge Enzyme-Like Proteins (BBLs) knockout were investigated. The knockout of BBLs has limited effect on tobacco agricultural traits. After the BBLs knockout, nicotine and most alkaloids are significantly reduced, but the content of myosmine and its derivatives increases dramatically. In order to identify the gene editing of tobacco, principal component analysis (PCA) was performed on the FT-IR and 2DCOS-IR spectroscopy data. The results showed that FT-IR can distinguish between tobacco roots and leaves but cannot classify the gene mutation tobacco from the wild one. 2DCOS-IR can enhance the characteristics of the samples due to the increased apparent resolution of the spectra. Using the autopeaks in the synchronous map for PCA analysis, we successfully identified the mutants with an accuracy of over 90%. |
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| Keywords: | gene-edited tobacco BBLs FT-IR 2DCOS-IR identification |
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