AUTHOR=Sun Heng , Song Heyun , Deng Xianbao , Liu Juan , Yang Dong , Zhang Minghua , Wang Yuxin , Xin Jia , Chen Lin , Liu Yanling , Yang Mei TITLE=Transcriptome-Wide Characterization of Alkaloids and Chlorophyll Biosynthesis in Lotus Plumule JOURNAL=Frontiers in Plant Science VOLUME=Volume 13 - 2022 YEAR=2022 URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2022.885503 DOI=10.3389/fpls.2022.885503 ISSN=1664-462X ABSTRACT=Lotus plumule is a green tissue in the middle of seeds which predominantly accumulate bisbenzylisoquinoline alkaloids (bis-BIAs) and chlorophyll (Chl). However, the biosynthetic mechanisms of these two metabolites remain largely unknown in lotus. This study used physiological and RNA sequencing (RNA-Seq) approaches to characterize the development and molecular mechanisms of bis-BIAs and Chl biosynthesis in lotus plumule. Physiological analysis revealed that exponential plumule growth occurred between 9-15 days after pollination (DAP), which coincided with the onset of bis-BIAs biosynthesis and its subsequent rapid accumulation. Transcriptome analysis of lotus plumule identified a total of 8,725 differentially expressed genes (DEGs), representing ~ 27.7% of all transcripts in the lotus genome. Sixteen structural DEGs potentially associated with bis-BIAs biosynthesis were identified. Of these, 12 encoded O-methyltransferases (OMTs) which are likely involved in the methylation and bis-BIAs diversity in lotus. In addition, functionally divergent paralogous and redundant homologous gene members of BIAs biosynthesis pathway as well as transcription factors co-expressed with bis-BIAs and Chl biosynthesis genes were identified. Twenty-two genes encoding 16 conserved enzymes of the Chl biosynthesis pathway were identified, with majority being significantly up-regulated by Chl biosynthesis. Photosynthesis and Chl biosynthesis pathways were simultaneously activated during lotus plumule development. Moreover, our results showed that light-driven Pchlide reduction is essential for Chl biosynthesis in lotus plumule. These results will be useful for enhancing our understanding of alkaloids and Chl biosynthesis in plants.