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.

Harvest yields depend on the plant's ability to fix carbon and deal with changing environmental conditions. Especially during seasonal and diurnal cycles, the plant must constantly adjust its metabolism according to available resources or external stressors. The metabolic changes that a plant undergoes in response to stress are well understood, but the long-distance signaling mechanisms that facilitate communication throughout the plant are less studied. The phloem is considered the predominant conduit for the bidirectional transport of these signals through metabolites, nucleic acids, proteins, and lipids. Lipid trafficking through the phloem in particular attracted our attention due to its reliance on soluble lipid-binding proteins (LBP) that generate and solubilize otherwise membrane-associated lipids. The Phloem Lipid-Associated Family Protein (PLAFP) from Arabidopsis thaliana is generated in response to abiotic stress as is its lipid-ligand phosphatidic acid (PA). PLAFP is proposed to transport PA through the phloem in response to drought stress. To understand the interactions between PLAFP and PA, almost 100 independent systems comprised of the protein and one PA, or a plasma membrane containing varying amounts of PA, were simulated. In the simulations, PLAFP does bind to the plasma membrane independent of the PA concentration, and it adopts a binding pose, where W41 and R82 penetrate the membrane surface and anchor PLAFP. This triggers a separation of the two loop regions containing W41 and R82. Subsequently, PA does insert into PLAFP's beta-sandwich and multiple amino acids besides W41 and R82 are identified that drive the insertion. Fine-tuning the protein-membrane and protein-PA interface by mutating a selection of these amino acids could allow modulating the signaling sensitivity to the climate the plant is supposed to grow in.