School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, People's Republic of China.
School of Pharmacy, Jiangsu University, Zhenjiang 212013, People's Republic of China.
J Agric Food Chem. 2021 Sep 22;69(37):10989-10998. doi: 10.1021/acs.jafc.1c03932. Epub 2021 Sep 13.
The alkaloid 1-deoxynojirimycin (DNJ) is one of the major bioactive compounds in mulberry leaves ( L.). Previously, we discovered four key genes involved in the pathway from lysine to piperidine in the biosynthesis of DNJ in mulberry leaves, (MG727866), (MH205733), (MT989445), and (MT989446), which encoded lysine decarboxylase, copper amine oxidase, and short-chain dehydrogenase/reductase 1 and 2, respectively. However, the functions of these four genes have not been verified yet. Here, these four genes were successfully cloned and used for the establishment of C58C1 mediated overexpression genetic transformation systems and GV3101 -mediated virus-induced gene silencing transformation systems in order to verify the influence of these four genes on the biosynthetic content of DNJ in mulberry leaves. The results showed that the content of DNJ increased after the four genes were overexpressed. When these four genes were silenced, the gene expression was blocked, which affected the biosynthesis of DNJ, and the DNJ content decreased. The above results indicated that these four genes participated in DNJ biosynthesis. This study provided a foundation for further elucidating the regulatory mechanisms of DNJ biosynthesis in mulberry leaves.
生物碱 1-脱氧野尻霉素（DNJ）是桑叶（L.）中主要的生物活性化合物之一。先前，我们发现了参与桑叶中 DNJ 生物合成途径从赖氨酸到哌啶的四个关键基因，即（MG727866）、（MH205733）、（MT989445）和（MT989446），它们分别编码赖氨酸脱羧酶、铜胺氧化酶和短链脱氢酶/还原酶 1 和 2。然而，这些四个基因的功能尚未得到验证。在这里，我们成功克隆了这四个基因，并用于建立 C58C1 介导的过表达遗传转化系统和 GV3101 介导的病毒诱导基因沉默转化系统，以验证这四个基因对桑叶中 DNJ 生物合成含量的影响。结果表明，四个基因过表达后，DNJ 的含量增加。当这四个基因沉默时，基因表达受阻，影响了 DNJ 的生物合成，DNJ 的含量下降。上述结果表明，这四个基因参与了 DNJ 的生物合成。本研究为进一步阐明桑叶中 DNJ 生物合成的调控机制提供了基础。
