Maize terpene synthase 1 impacts insect behavior via the production of monoterpene volatiles ß-myrcene and linalool.

Plant-derived volatiles are important mediators of plant-insect interactions as they can provide cues for host location and quality, or act as direct or indirect defense molecules. The volatiles produced by Zea mays (maize) include a range of terpenes, likely produced by several of the terpene synthases (TPS) present in maize. Determining the roles of specific terpene volatiles and individual TPSs in maize-insect interactions is challenging due to the promiscuous nature of TPSs in vitro and their potential for functional redundancy. In this study, we used metabolite GWAS of a sweetcorn diversity panel infested with Spodoptera frugiperda (fall armyworm) to identify genetic correlations between TPSs and individual volatiles. This analysis revealed a correlation between maize terpene synthase 1 (ZmTPS1) and emission of the monoterpene volatiles linalool and ß-myrcene. Electroantennogram assays showed gravid S. frugiperda could detect both linalool and ß-myrcene. Quantification of headspace volatiles in a maize tps1 loss-of-function mutant confirmed that ZmTPS1 is an important contributor to linalool and ß-myrcene emission in maize. Furthermore, pairwise choice assays between tps1 mutant and wild-type plants showed that ZmTPS1, and by extension its volatile products, aid host location in the chewing insect S. frugiperda, yet repel the sap-sucking pest, Rhopalosiphum maidis (corn leaf aphid). On the other hand, ZmTPS1 had no impact on indirect defense via the recruitment of the parasitoid Cotesia marginiventris. ZmTPS1 is therefore an important mediator of the interactions between maize and its insect pests.

The impact of climate change on maize chemical defenses.

Climate change is increasingly affecting agriculture, both at the levels of crops themselves, and by altering the distribution and damage caused by insect or microbial pests. As global food security depends on the reliable production of major crops such as maize (Zea mays), it is vital that appropriate steps are taken to mitigate these negative impacts. To do this a clear understanding of what the impacts are and how they occur is needed. This review focuses on the impact of climate change on the production and effectiveness of maize chemical defenses, including volatile organic compounds, terpenoid phytoalexins, benzoxazinoids, phenolics, and flavonoids. Drought, flooding, heat stress, and elevated concentrations of atmospheric carbon dioxide, all impact the production of maize chemical defenses, in a compound and tissue-specific manner. Furthermore, changes in stomatal conductance and altered soil conditions caused by climate change can impact environmental dispersal and effectiveness certain chemicals. This can alter both defensive barrier formation and multitrophic interactions. The production of defense chemicals is controlled by stress signaling networks. The use of similar networks to co-ordinate the response to abiotic and biotic stress can lead to complex integration of these networks in response to the combinatorial stresses that are likely to occur in a changing climate. The impact of multiple stressors on maize chemical defenses can therefore be different from the sum of the responses to individual stressors and challenging to predict. Much work remains to effectively leverage these protective chemicals in climate-resilient maize.

Production of the Green Leaf Volatile (Z)-3-Hexenal by a Zea mays Hydroperoxide Lyase.

Plant-produced volatile compounds play important roles in plant signaling and in the communication of plants with other organisms. Many plants emit green leaf volatiles (GLVs) in response to damage or attack, which serve to warn neighboring plants or attract predatory or parasitic insects to help defend against insect pests. GLVs include aldehydes, esters, and alcohols of 6-carbon compounds that are released rapidly following wounding. One GLV produced by maize (Zea mays) is the volatile (Z)-3-hexenal; this volatile is produced from the cleavage of (9Z,11E,15Z)-octadecatrienoic acid by hydroperoxide lyases (HPLs) of the cytochrome P450 CYP74B family. The specific HPL in maize involved in (Z)-3-hexenal production had not been determined. In this study, we used phylogenetics with known HPLs from other species to identify a candidate HPL from maize (ZmHPL). To test the ability of the putative HPL to produce (Z)-3-hexenal, we constitutively expressed the gene in Arabidopsis thaliana ecotype Columbia-0 that contains a natural loss-of-function mutant in AtHPL and examined the transgenic plants for restored (Z)-3-hexenal production. Volatile analysis of leaves from these transgenic plants showed that they did produce (Z)-3-hexenal, confirming that ZmHPL can produce (Z)-3-hexenal in vivo. Furthermore, we used gene expression analysis to show that expression of ZmHPL is induced in maize in response to both wounding and the insect pests Spodoptera frugiperda and Spodoptera exigua. Our study demonstrates that ZmHPL can produce GLVs and highlights its likely role in (Z)-3-hexenal production in response to mechanical damage and herbivory in maize.

Dedicated farnesyl diphosphate synthases circumvent isoprenoid-derived growth-defense tradeoffs in Zea mays.

Zea mays (maize) makes phytoalexins such as sesquiterpenoid zealexins, to combat invading pathogens. Zealexins are produced from farnesyl diphosphate in microgram per gram fresh weight quantities. As farnesyl diphosphate is also a precursor for many compounds essential for plant growth, the question arises as to how Z. mays produces high levels of zealexins without negatively affecting vital plant systems. To examine if specific pools of farnesyl diphosphate are made for zealexin synthesis we made CRISPR/Cas9 knockouts of each of the three farnesyl diphosphate synthases (FPS) in Z. mays and examined the resultant impacts on different farnesyl diphosphate-derived metabolites. We found that FPS3 (GRMZM2G098569) produced most of the farnesyl diphosphate for zealexins, while FPS1 (GRMZM2G168681) made most of the farnesyl diphosphate for the vital respiratory co-factor ubiquinone. Indeed, fps1 mutants had strong developmental phenotypes such as reduced stature and development of chlorosis. The replication and evolution of the fps gene family in Z. mays enabled it to produce dedicated FPSs for developmentally related ubiquinone production (FPS1) or defense-related zealexin production (FPS3). This partitioning of farnesyl diphosphate production between growth and defense could contribute to the ability of Z. mays to produce high levels of phytoalexins without negatively impacting its growth.

Zea mays Volatiles that Influence Oviposition and Feeding Behaviors of Spodoptera frugiperda.

Fall armyworm (Spodoptera frugiperda) is a major global pest of many crops, including maize (Zea mays). This insect is known to use host plant-derived volatile organic compounds to locate suitable hosts during both its adult and larval stages, yet the function of individual compounds remains mostly enigmatic. In this study, we use a combination of volatile profiling, electrophysiological assays, pair-wise choice behavioral assays, and chemical supplementation treatments to identify and assess specific compounds from maize that influence S. frugiperda host location. Our findings reveal that methyl salicylate and (E)-alpha-bergamotene are oviposition attractants for adult moths but do not impact larval behavior. While geranyl acetate can act as an oviposition attractant or repellent depending on the host volatile context and (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT) is an oviposition deterrent. These compounds can also be attractive to the larvae when applied to specific maize inbreds. These data show that S. frugiperda uses different plant volatile cues for host location in its adult and larval stage and that the background volatile context that specific volatiles are perceived in, alters their impact as behavioral cues.

Phenotypic characterization of Arabidopsis thaliana lines overexpressing AVP1 and MIOX4 in response to abiotic stresses.

PREMISE: AVP1 (H+-pyrophosphatase) and MIOX4 (myo-inositol oxygenase) are genes that, when overexpressed individually, enhance the growth and abiotic stress tolerance of Arabidopsis thaliana plants. We propose that pyramiding AVP1 and MIOX4 genes will further improve stress tolerance under water-limited and salt-stress conditions. METHODS: MIOX4 and AVP1 reciprocal crosses were developed and phenomic approaches used to investigate the possible synergy between these genes. RESULTS: Under normal and stress conditions, the crosses had higher foliar ascorbate content than the wild-type and parental lines. Under water-limited conditions, the crosses also displayed an enhanced growth rate and biomass compared with the control. The observed increases in photosystem II efficiency, linear electron flow, and relative chlorophyll content may have contributed to this observed phenotype. Additionally, the crosses retained more water than the controls when subjected to salt stress. Higher seed yields were also observed in the crosses compared with the controls when grown under salt and water-limitation stresses. DISCUSSION: Overall, these results suggest the combinatorial effect of overexpressing MIOX4 and AVP1 may be more advantageous than the individual traits for enhancing stress tolerance and seed yields during crop improvement.

Mechanisms underlying the enhanced biomass and abiotic stress tolerance phenotype of an Arabidopsis MIOX over-expresser.

Myo-inositol oxygenase (MIOX) is the first enzyme in the inositol route to ascorbate (L-ascorbic acid, AsA, vitamin C). We have previously shown that Arabidopsis plants constitutively expressing MIOX have elevated foliar AsA content and displayed enhanced growth rate, biomass accumulation, and increased tolerance to multiple abiotic stresses. In this work, we used a combination of transcriptomics, chromatography, microscopy, and physiological measurements to gain a deeper understanding of the underlying mechanisms mediating the phenotype of the AtMIOX4 line. Transcriptomic analysis revealed increased expression of genes involved in auxin synthesis, hydrolysis, transport, and metabolism, which are supported by elevated auxin levels both in vitro and in vivo, and confirmed by assays demonstrating their effect on epidermal cell elongation in the AtMIOX4 over-expressers. Additionally, we detected up-regulation of transcripts involved in photosynthesis and this was validated by increased efficiency of the photosystem II and proton motive force. We also found increased expression of amylase leading to higher intracellular glucose levels. Multiple gene families conferring plants tolerance/expressed in response to cold, water limitation, and heat stresses were found to be elevated in the AtMIOX4 line. Interestingly, the high AsA plants also displayed up-regulation of transcripts and hormones involved in defense including jasmonates, defensin, glucosinolates, and transcription factors that are known to be important for biotic stress tolerance. These results overall indicate that elevated levels of auxin and glucose, and enhanced photosynthetic efficiency in combination with up-regulation of abiotic stresses response genes underly the higher growth rate and abiotic stresses tolerance phenotype of the AtMIOX4 over-expressers.

DNA barcoding and TLC as tools to properly identify natural populations of the Mexican medicinal species Galphimia glauca Cav.

Galphimia glauca is a plant that is endemic to Mexico and has been commonly used since pre-Hispanic times to treat various illnesses, including central nervous system disorders and inflammation. The first studies investigating a natural population of G. glauca in Mexico showed that the plant has anxiolytic and sedative activities in mice and humans. The plant's bioactive compounds were isolated and identified, and they belong to a family of nor-secofriedelanes called galphimines. The integration of DNA barcoding and thin-layer chromatography analysis was performed to clarify whether the botanical classification of the populations in the study, which were collected in different regions of Mexico, as G. glauca was correct or if the populations consist of more than one species of the genus Galphimia. We employed six DNA barcodes (matK, rbcL, rpoC1, psbA-trnH, ITS1 and ITS2) that were analyzed individually and in combination and then compared each other, to indicate differences among the studied populations. In the phylogenetic analysis, ITS1 and ITS2 markers as well as the combination of all DNA regions were the most efficient for discriminating the population studied. The thin-layer chromatography analysis exhibited four principal chemical profiles, one of which corresponded to the populations that produced galphimines. DNA barcoding was consistent and enabled us to differentiate the populations that produce galphimines from those that do not. The results of this investigation suggest that the studied populations belong to at least four different species of the genus Galphimia. The phylogenetic analysis and the thin-layer chromatography chemical profiles were convenient tools for establishing a strong relationship between the genotype and phenotype of the studied populations and could be used for quality control purposes to prepare herbal medicines from plants of the genus Galphimia.