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Abstract
Jasmonic acid (JA) carboxyl methyltransferase (JMT), a key enzyme in jasmonate-regulated plant responses, may be involved in plant defense and development by methylating JA to MeJA, thus influencing the concentrations of MeJA in plant. In this study, we isolated the JMT gene from Salvia miltiorrhiza, an important medicinal plant widely used to treat cardiovascular disease. We present a genetic manipulation strategy to enhance the production of phenolic acids by overexpresion SmJMT in S. miltiorrhiza. Global transcriptomic analysis using RNA sequencing showed that the expression levels of genes involved in the biosynthesis pathway of phenolic acids and MeJA were upregulated in the overexpression lines. In addition, the levels of endogenous MeJA, and the accumulation of rosmarinic acid (RA) and salvianolic acid (Sal B), as well as the concentrations of total phenolics and total flavonoids in transgenic lines, were significantly elevated compared with the untransformed control. Our results demonstrate that overexpression of SmJMT promotes the production of phenolic acids through simultaneously activating genes encoding key enzymes involved in the biosynthesis pathway of phenolic acids and enhancing the endogenous MeJA levels in S. miltiorrhiza.
Highlights
Salvia miltiorrhiza Bunge (Lamiaceae) is a well-known traditional Chinese herb with significant medicinal and economic value
As described above, overexpressing jasmonic acid carboxyl methyltransferase (JMT) in plants elevated the level of endogenous methyl jasmonate (MeJA), and transgenic plants exhibited constitutive expression of jasmonate-responsive genes [21], while most genes involved in the biosynthesis of salvianolic acid B (Sal B) and rosmarinic acid (RA) could be induced by jasmonates in S. miltiorrhiza [8,9,28,32,33]
To determine the evolutionary relationship of the SmJMT with the members of JMTs from other species, an unrooted phylogenetic tree was constructed using the amino-acid sequences of S. miltiorrhiza JMT and 27 JMTs from other species (Figure 2)
Summary
Salvia miltiorrhiza Bunge (Lamiaceae) is a well-known traditional Chinese herb with significant medicinal and economic value. Jasmonates which include jasmonic acid (JA), methyl jasmonate (MeJA), and its cyclopentane derivatives, are a class of plant hormone that regulate many aspects of plant development such as root growth, production of viable pollen, fruit ripening, and senescence [11,12] They are involved in plant responses to biotic and abiotic stresses including insect attack, wounding, water deficiency, ultraviolet (UV) light, pathogen infection, and ozone [13,14,15]. As described above, overexpressing JMT in plants elevated the level of endogenous MeJA, and transgenic plants exhibited constitutive expression of jasmonate-responsive genes [21], while most genes involved in the biosynthesis of Sal B and RA could be induced by jasmonates in S. miltiorrhiza [8,9,28,32,33]. The contents of RA, Sal B, total phenols, total flavonoids, and endogenous MeJA in transgenic and control lines were analyzed using different biological techniques
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