Journal: Horticulture Research

Article Title: Study on biosynthesis pathway and accumulation mechanism of the dihydrochalcones in Lithocarpus litseifolius

doi: 10.1093/hr/uhag061

Figure Lengend Snippet: In vitro biosynthesis of phloretin from dihydro- p -coumaraldehyde (dihydro-pCAld) or dihydro- p -cinnamic acid (dihydro-pCAci) catalyzed by LlALDH1, Ll4CL2 and LlCHS1, or LlCCR and LlCHS1. (a) Dihydro-pCAld, dihydro-pCAci and phloretin standards on HPLC chromatogram, as well as the extracted ion chromatogram (EIC) and MS signal spectrum of phloretin. (b–h) HPLC chromatograms, EICs, and MS spectra of the catalytic products in different reaction systems; (b) Reaction system 1: including the substrates dihydro- p -coumaroyl-CoA (dihydro-pCCoA) and malonyl-CoA (MCoA), and the enzyme LlCHS1; (c) Whole reaction system 2: including the substrates dihydro-pCAci and MCoA, cofactors MgCl 2 , ATP and CoA, as well as the enzymes Ll4CL2 and LlCHS1; (d) Whole reaction system 3: including the substrates dihydro-pCAld and MCoA, cofactors NAD + , MgCl 2 , ATP and CoA, as well as the enzymes LlALDH1, Ll4CL2 and LlCHS1; (e) Reaction system 2 without Ll4CL2; (f) Reaction system 3 without Ll4CL2; (g) Reaction system 3 without LlALDH1; (h) Reaction system 3 without LlALDH1 and Ll4CL2; (i) Whole reaction system 4: including the substrates dihydro-pCAld and MCoA, cofactors NADP + and CoA, and the enzymes LlCCR and LlCHS1; (j) Reaction system 4 without LlCCR; (k) Potential biosynthetic step of phloretin as well as the required enzymes, substrates, and cofactors. Peak 1, dihydro-pCAci; peak 2, dihydro-pCAld; peak 3, dihydro-CTAL (dihydro- p -coumaroyl triacetic acid lactone); peak 4, dihydro-BNY (dihydro-bisnoryangonin); peak 5, phloretin.

Article Snippet: pCCoA, FCoA, SCoA, pCAld, CAld, SAld, dihydro-pCAld, pCAci, dihydro-pCAci, FAci, SAci, phloretin, phlorizin, trilobatin, phenylmethanesulfonylfluoride (PMSF), NADP + , NAD + , NADPH, CoA, and ATP were purchased from Shanghai Yuanye Biotechnology Co., Ltd (Shanghai, China).

Techniques: In Vitro

Journal: Horticulture Research

Article Title: Study on biosynthesis pathway and accumulation mechanism of the dihydrochalcones in Lithocarpus litseifolius

doi: 10.1093/hr/uhag061

Figure Lengend Snippet: High yield of phloretin from dihydro- p -coumaraldehyde (dihydro-pCAld) or dihydro- p -cinnamic acid (dihydro-pCAci) catalyzed by LlALDH1, Ll4CL2 and LlCHS1 or LlCCR and LlCHS1 in reaction solutions without cofactor CoA. (a-c) HPLC chromatograms, extracted ion chromatograms (EICs), and MS spectra of the catalytic products in different reaction systems. (a) Reaction system 2 without CoA (including the substrates dihydro-pCAci and malonyl-CoA (MCoA), cofactors MgCl 2 and ATP, as well as the enzymes Ll4CL2 and LlCHS1). (b) Reaction system 3 without CoA (including the substrates dihydro-pCAld and MCoA, cofactors NAD + , MgCl 2 and ATP, as well as the enzymes LlALDH1, Ll4CL2 and LlCHS1); (c) Reaction system 4 without CoA (including the substrates dihydro-pCAld and MCoA, and cofactor NADP + , and the enzymes LlCCR and LlCHS1). Peak 1, dihydro-pCAci; peak 2, dihydro-pCAld; peak 3, dihydro-CTAL (dihydro- p -coumaroyl triacetic acid lactone); peak 4, dihydro-BNY (dihydro-bisnoryangonin); peak 5, phloretin.

Article Snippet: pCCoA, FCoA, SCoA, pCAld, CAld, SAld, dihydro-pCAld, pCAci, dihydro-pCAci, FAci, SAci, phloretin, phlorizin, trilobatin, phenylmethanesulfonylfluoride (PMSF), NADP + , NAD + , NADPH, CoA, and ATP were purchased from Shanghai Yuanye Biotechnology Co., Ltd (Shanghai, China).

Techniques:

Journal: bioRxiv

Article Title: Interactomics-driven discovery of alkaloid biosynthetic pathways in kratom

doi: 10.64898/2026.05.01.722234

Figure Lengend Snippet: a) Transcriptional profiles of 60 positive interactors and the upstream MIA-pathway genes across 49 published kratom transcriptomes were analyzed, and genes exhibiting co-expression patterns are shown. Each column represents a biological replicate, and each row corresponds to a distinct gene. Expression levels are shown as Z-scores calculated from log 2 -normalized TPM values, ranging from −1.4 (blue, lower expression) to +5.1 (red, higher expression). Among the 60 putative interacting MsMDRs, three functional and interacting MsMDRs, such as MsMDR141, MsMDR290, and MsDCS1, were highly expressed in roots from samples with high mitragynine levels (Root 1) and young leaves. Other genes highly expressed include MsSLS, MsSTR, Ms7DLGT, MsTDC, MsCPR, Ms8HGO, MsLAMT1, and MsIO in the strictosidine biosynthetic pathway. Sample descriptors include: Root 1 (high-mitragynine), Root 2 (low-mitragynine), Leaf 2 (leaf bract), and Leaf 3 (leaf after wounding). b) Genomic mapping of the three co-expressed MsMDRs identified a 320-kb biosynthetic gene cluster in Tig00011620 scaffold. Green: previously reported MsMDRs, including MsDCS1, 3, 4, a truncated version of MsDCS3, and MsMDR4; Red: newly discovered functional MsMDRs, including MsMDR141 (three isogenes), MsMDR290, and MsMDR320. Black: putative interacting MsMDRs that did not exhibit activity in biochemical screening. Abbreviations: MDR, medium-chain dehydrogenase/reductase; SLS, secologanin synthase; STR, strictosidine synthase; 7-DLGT, 7-deoxyloganetic acid-O-glucosyl transferase; TDC, tryptophan decarboxylase; DCS, dihydrocorynantheine synthase; CPR, NADPH-cytochrome P450 reductase; 8-HGO, 8-hydroxygeraniol oxidoreductase; LAMT, loganic acid O-methyltransferase; IO, 7-deoxyloganetic acid synthase/iridoid oxidase.

Article Snippet: Dextrose, yeast extract (YE), peptone, Luria-Bertani (LB) broth, LB agar, Terrific broth (TB), agar, acetonitrile, formic acid, imidazole, β-nicotinamide adenine dinucleotide phosphate reduced tetrasodium salt (NADPH) and protease inhibitor (100x) were purchased from Thermo Fisher Scientific.

Techniques: Expressing, Gene Expression, Functional Assay, Activity Assay