JRE4 is a master transcriptional regulator of defense-related steroidal glycoalkaloids in tomato.

Steroidal glycoalkaloids (SGAs) are specialized anti-nutritional metabolites that accumulate in Solanum lycopersicum (tomato) and Solanum tuberosum (potato). A series of SGA biosynthetic genes is known to be upregulated in Solanaceae species by jasmonate-responsive Ethylene Response Factor transcription factors, including JRE4 (otherwise known as GAME9), but the exact regulatory significance in planta of each factor has remained unaddressed. Here, via TILLING-based screening of an EMS-mutagenized tomato population, we isolated a JRE4 loss-of-function line that carries an amino acid residue missense change in a region of the protein important for DNA binding. In this jre4 mutant, we observed downregulated expression of SGA biosynthetic genes and decreased SGA accumulation. Moreover, JRE4 overexpression stimulated SGA production. Further characterization of jre4 plants revealed their increased susceptibility to the generalist herbivore Spodoptera litura larvae. This susceptibility illustrates that herbivory resistance is dependent on JRE4-mediated defense responses, which include SGA accumulation. Ethylene treatment attenuated the jasmonate-mediated JRE4 expression induction and downstream SGA biosynthesis in tomato leaves and hairy roots. Overall, this study indicated that JRE4 functions as a primary master regulator of SGA biosynthesis, and thereby contributes toward plant defense against chewing insects.

Jasmonate-induced biosynthesis of steroidal glycoalkaloids depends on COI1 proteins in tomato.

In tomato, perception of jasmonates by a receptor complex, which includes the F-box protein CORONATINE INSENSITIVE 1 (COI1), elicits biosynthesis of defensive steroidal glycoalkaloids (SGAs) via a jasmonate-responsive ERF transcription factor, JRE4/GAME9. Although JRE4 is upregulated by jasmonate and induces the expression of many metabolic genes involved in SGA biosynthesis, it is not known whether JRE4 alone is sufficient for increased SGA biosynthesis upon activation of jasmonate signaling. Here, we show that application of methyl jasmonate induces the expression of JRE4 and SGA biosynthesis genes in leaves and hairy roots of wild-type tomato, but not in jasmonic acid insensitive 1 (jai1), a loss-of-function mutant allele of the tomato COI1 gene. Induced overexpression of JRE4 increased the expression of SGA biosynthesis genes in transgenic hairy roots of both wild-type tomato and the jai1 mutant, suggesting that JRE4 is the primary transcription factor that functions downstream of the jasmonate signaling pathway.

Eco-friendly approach to decrease the harmful effects of untreated wastewater on growth, yield, biochemical constituents, and heavy metal contents of carrot (Daucus carota L.).

Here, the impact of irrigation using untreated wastewater (WW) on carrots (Daucus carota L.) was examined. We hypothesized that the addition of ethylenediaminetetraacetic acid (EDTA), dry algal powder (Spirulina platensis or Chlorella vulgaris), and Salix alba leaves powder would function as chelators for harmful contaminants in wastewater. The findings showed that irrigation of carrot plants with the sampled untreated wastewater led to significant decreases in the shoot lengths, fresh, dry weights of shoots and roots at stage I, the diameter of roots, pigment content, carotenoids, total soluble carbohydrate content, and soluble protein content. Furthermore, a significantly increased level of proline, total phenols, and the activities of polyphenol oxidase (PPO), peroxidase (POX), superoxide dismutase (SOD), and catalase (CAT) was identified in stage I samples. In contrast to the stage I, the length of the roots, the number of leaves on each plant, wet and dry weights of the stage II roots were all greatly enhanced. In spite of the increased yield due to the wastewater irrigation, carrot roots irrigated with wastewater had significantly more cadmium (Cd), nickel (Ni), cobalt (Co), and lead (Pb) than is considered safe. Our data clearly show that the application of Spirulina platensis, Chlorella vulgaris, EDTA, and leaves powder of salix was able to alleviate the toxicity of wastewater on carrot plants. For example, we recorded a significant decrease in the accumulation of carrot's Cd, Ni, Co, and Pb contents. We conclude that the treatments with Spirulina platensis and Chlorella vulgaris can be utilized as eco-friendly tools to lessen the damaging effects of wastewater irrigation on carrot plants.

Identification of genes regulated by a jasmonate- and salt-inducible transcription factor JRE3 in tomato.

In Solanum lycoperisicum (tomato), a transcription factor of APETALA2/ETHYLENE RESPONSE FACTOR (AP2/ERF) family, JASMONATE-RESPONSIVE ERF 3 (JRE3), is a closest homolog of JRE4, a master transcriptional regulator of steroidal glycoalkaloid (SGA) biosynthesis. In tomato genome, JRE3 resides in a gene cluster with JRE4 and related JRE1, JRE2, and JRE5, while JRE6 exists as a singleton on a different chromosome. All of the JREs are induced by jasmonates (JAs), whereas sodium chloride (NaCl) treatment drastically increases the expression of the JREs except for JRE4 and JRE6. In this study, to get insights into the regulatory function of the JA- and NaCl-inducible JRE3, a series of genes upregulated by ß-estradiol-induced overexpression of JRE3 are identified with microarray analysis in transgenic tomato hairy roots. No gene involved in the SGA pathway has been identified through the screening, confirming the functional distinction between JRE3 and JRE4. Among the JRE3-regulated genes, we characterize the stress-induced expression of genes encoding malate synthase and tonoplast dicarboxylate transporter both involved in malate accumulation. In transient transactivation assay, we reveal that both terminal regions of JRE4, but not a central DNA-binding domain, are indispensable for the induction of a gene involved in the JRE4 regulon. Functional differentiation of the JREs is discussed.