Plant Science

This is the supporting data related to our manuscript.

Supplementary Files 1 and 2. Scripts for analyzing images. Resulting data.

Cannabinoids are terpenophenolic compounds produced by Cannabis sativa L., which accumulate in storage cavities of glandular trichomes as a part of the exudates. We investigated if tetrahydrocannabinolic acid synthase and cannabidiolic acid synthase, which are involved in the last step of cannabinoid biosynthesis, are also secreted into Cannabis trichome exudates. The exudates were collected by microsuction from storage cavities of Cannabis glandular trichomes and were subjected for proteomic and metabolomic analyses. The catalytic activity of the exudates was documented by cannabigerolic acid biotransformation studies under hydrophobic conditions. Electrophoretic separations revealed protein bands at ~65 kDa, which were further identified as tetrahydrocannabinolic acid synthase and cannabidiolic acid synthase. The accumulation of the enzymes in trichome exudates increased substantially during the flowering period in the drug-type Cannabis plants. The content of cannabinoids increased significantly after incubating hexane-diluted trichome exudates with cannabigerolic acid. In this study, we showed that Cannabis glandular trichomes secrete and accumulate cannabinoid synthases in storage cavities, and are able to convert cannabigerolic acid under hydrophobic trichome-mimicking conditions. Metabolite profiling of the exudates revealed compounds with hydrophilic, osmoprotective and amphiphilic properties, which may play a role in providing a necessary aqueous microenvironment, which enables enzyme solubility and biocatalysis under hydrophobic conditions of glandular trichomes.

Raw data for plotting Figs. 2 and 5.

We would like to submit the enclosed manuscript entitled " Molecular characterization and expression analysis of the critical floral gene MdAGL24-like in red-fleshed apple", which we wish to be considered for publication in Plant Science.. Compared with annual herbaceous plants, flowering is the premise of fruit bearing. The flowering of woody apple trees is relatively late because of its long juvenile phase that greatly affects the genetic improvement process of these plants. To better understand the molecular regulation of floral transition and flower development in red-fleshed apple, we isolated and characterized a floral MADS-box gene, MdAGL24-like, in this paper. MdAGL24-like shares sequence similarity with AGAMOUS-LIKE 24 (AGL24) from other species. It’s dynamically expressed in flowers, followed by roots and fruits. Like other transcript factors, MdAGL24-like was localized in the nucleus. MdAGL24-like could interact with MdSOC1 and MdAP1 in vivo and in vitro. MdAGL24-like and MdSOC1 could increase each other’s expression by binding the CArG motifs in their promoters. Unlike MdSOC1, MdAGL24-like couldn’t directly bind the downstream gene MdLFY. Ectopic overexpression of MdAGL24-like in wild-type Arabidopsis induced early flowering similar to the phenotypes induced by other AGL24 genes. Like AGL24 in Arabidopsis, MdAGL24-like could somewhat rescue the late-flowering phenotype of agl24. These results would clarify the molecular mechanism underlying flowering and provide a method of shortening the juvenile period in red-fleshed apples and other fruit trees. The work described has not been submitted elsewhere for publication, in whole or in part. All authors have seen the manuscript and approved to submit to your journal. Thank you very much for your attention and consideration.

Blue Fluorescent Banana samples (leaves and fruits)

Supplemental data delineating the identifiers of each of the genes referenced in the phylogenetic tree figure.