经 CRISPR-Cas9 编辑的低尼古丁含量非转基因烟草的农艺性状、生物碱分析、FT-IR 和 2DCOS-IR 光谱鉴定。
The Agronomic Traits, Alkaloids Analysis, FT-IR and 2DCOS-IR Spectroscopy Identification of the Low-Nicotine-Content Nontransgenic Tobacco Edited by CRISPR-Cas9.
机构信息
Yunnan Key Laboratory of Tobacco Chemistry, Research and Development Center, China Tobacco Yunnan Industrial Co., Ltd., Kunming 650231, China.
Department of Chemistry, Tsinghua University, Beijing 100084, China.
出版信息
Molecules. 2022 Jun 14;27(12):3817. doi: 10.3390/molecules27123817.
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%.
本研究探讨了敲除小檗碱桥连酶样蛋白（BBLs）后烟草的农艺性状、生物碱含量以及傅里叶变换红外光谱（FT-IR）和二维相关红外光谱（2DCOS-IR）分析。BBLs 的敲除对烟草农艺性状的影响有限。敲除 BBLs 后，尼古丁和大多数生物碱显著减少，但肌蕈碱及其衍生物的含量显著增加。为了鉴定烟草的基因编辑，对 FT-IR 和 2DCOS-IR 光谱数据进行了主成分分析（PCA）。结果表明，FT-IR 可以区分烟草的根和叶，但不能将基因突变的烟草与野生烟草区分开来。2DCOS-IR 可以通过增加光谱的表观分辨率来增强样品的特征。使用同步图谱中的自峰进行 PCA 分析，我们成功地识别出突变体的准确率超过 90%。
Zotero 插件