Fig 1.
Effects of MeJA application (250 μmol L-1) on gentiopicroside biosynthesis in Gentiana macrophylla seedlings.
(A). Chromatogram of extraction from seedlings not exposed to MeJA. (B). Chromatogram of extraction from seedlings after 5 d of MeJA treatment. (C). Chromatogram of gentiopicroside standard. (D). Changes over time in gentiopicroside concentrations in treated seedlings relative to levels in untreated control (C) plants.
Table 1.
Overview of sequencing and assembly.
Fig 2.
COG classifications of assembled unigenes.
Fig 3.
GO classification of assembled unigenes.
Table 2.
Secondary metabolism pathways in Gentiana macrophylla sequencing.
Fig 4.
KEGG pathway enrichment analysis of DEGs.
Table 3.
DEGs up-regulated in JA biosynthesis and JA signaling pathway in M5 samples compared with expression in C samples.
Fig 5.
DEGs involved in alpha-linolenic acid metabolism in M5 samples compared with C samples.
3.1.1.4, secretory phospholipase A2; 1.13.11.12, lipoxygenase; 4.2.1.92, allene oxide synthase; 1.1.1.1, alcohol dehydrogenase; 5.3.99.6, allene oxide cyclase; 1.3.1.42, 12-oxophytodienoic acid reductase; OPCL1, OPC-8:0 CoA ligase 1; ACX, acyl-CoA oxidase; 2.3.1.16, acetyl-CoA acyltransferase 1; 2.1.1.141, jasmonate O-methyltransferase. Green box, putative encoding gene is down-regulated; red boxes, putative encoding genes are up-regulated; blue boxes, some putative encoding genes are up-regulated while others are down-regulated.
Table 4.
Putative genes that encode enzymes involved in gentiopicroside biosynthesis and DEGs for which expression is altered in response to MeJA treatment.
Fig 6.
DEGs involved in gentiopicroside biosynthesis pathway in M5 samples compared with C samples.
DXS, 1-deoxy-D-xylulose-5-phosphate synthase; DXR, deoxylulose 5-phosphate reductoisomerase; MCT, 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase; CMK, 4-(cytidine -5’-diphospho)- 2-C-methyl-D- erythritol kinase; MCS, 2-C-methyl-D-erythritol-2,4- cyclodiphosphate synthase; HDS, 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate synthase; HDR, 1-hydroxy-2-methyl -2-(E)-butenyl 4-diphosphate reductase; IDI, isopentenyl diphosphate isomerase; AACT, acetyl-CoA C-acetyltransferase; HMGS, 3-hydroxy-3-methylglutaryl-coenzyme A synthase; HMGR, 3-hydroxy-3- methylglutaryl-coenzyme A reductase; MK, mevalonate kinase; PMK, 5-phosphomevelonate kinase; MVD, mevalonate-5-pyrophosphate decarboxylase; GPPS, geranyl diphosphate synthase; GES, geraniol synthase; G10H, geraniol 8-oxidase/geraniol 10-hydroxylase; CPR, cytochrome P450 reductase; 8/10HGO, 8-hydroxygeraniol oxidoreductase; IS, iridoid synthase; IO, iridoid oxidase; 7-DLGT, 7-deoxyloganetic acid glucosyl transferase; 7-DLH, 7-deoxyloganic acid hydroxylase; LAMT, loganic acid O-methyltransferase; SLS, secologanin synthase. Red squares, putative encoding genes are up-regulated; blue squares, putative encoding genes are down-regulated.
Fig 7.
DEGs involved in glycolysis in M5 samples compared with C samples.
5.4.2.2, phosphoglucomutase; 2.7.1.1, hexokinase; 5.1.3.15, glucose-6-phosphate 1-epimerase; 4.1.2.13, fructose-bisphosphate aldolase; 5.3.1.1, triosephosphate isomerase; 1.2.1.12, glyceraldehyde 3-phosphate dehydrogenase; 2.7.2.3, phosphoglycerate kinase; 5.4.2.12, 2,3-bisphosphoglycerate- independent phosphoglycerate mutase; 4.2.1.11, enolase; 2.7.1.40, pyruvate kinase; 4.1.1.49, phosphoenolpyruvate carboxykinase; 1.2.4.1, pyruvate dehydrogenase E1 component alpha subunit; 4.1.1.1, pyruvate decarboxylase; 2.3.1.12, dihydrolipoamide acetyltransferase; 1.1.1.27, L-lactate dehydrogenase; 1.8.1.4, dihydrolipoamide dehydrogenase; 1.2.1.3, aldehyde dehydrogenase; 1.1.1.1, alcohol dehydrogenase. Green box, putative encoding gene is down-regulated; red boxes, putative encoding genes are up-regulated; blue boxes, some putative encoding genes are up-regulated while others are down-regulated.
Fig 8.
qRT-PCR validation of 12 unigenes involved in gentiopicroside biosynthesis in G. macrophylla seedlings.
DXS, 1-deoxy-D-xylulose-5-phosphate synthase; DXR, deoxylulose 5-phosphate reductoisomerase; GES, geraniol synthase; G10H, geraniol 8-oxidase/geraniol 10-hydroxylase; 8/10HGO, 8/10-hydroxygeraniol oxidoreductase; HMGR, 3-hydroxy-3-methylglutaryl-coenzyme A reductase; HMGS, 3-hydroxy-3-methylglutaryl-coenzyme A synthase; MVD, mevalonate-5- pyrophosphate decarboxylase; CPR, cytochrome P450 reductase; HDS, 1-hydroxy-2-methyl-2- (E)-butenyl 4-diphosphate synthase; HDR, 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase.