Creation of a Plant Metabolite Spectral Library for Untargeted and Targeted Metabolomics.

Large-scale high throughput metabolomic technologies are indispensable components of systems biology in terms of discovering and defining the metabolite parts of the system. However, the lack of a plant metabolite spectral library limits the metabolite identification of plant metabolomic studies. Here, we have created a plant metabolite spectral library using 544 authentic standards, which increased the efficiency of identification for untargeted metabolomic studies. The process of creating the spectral library was described, and the mzVault library was deposited in the public repository for free download. Furthermore, based on the spectral library, we describe a process of creating a pseudo-targeted method, which was applied to a proof-of-concept study of Arabidopsis leaf extracts. As authentic standards become available, more metabolite spectra can be easily incorporated into the spectral library to improve the mzVault package.

Differential tolerance to heat stress of young leaves compared to mature leaves of whole plants relate to differential transcriptomes involved in metabolic adaptations to stress.

Plants respond to heat shock by regulating gene expression. While transcriptomic changes in response to heat stress are well studied, it is not known whether young and old leaves reprogram transcription differently upon stress. When whole plants of Arabidopsis thaliana were subjected to heat shock, young leaves were affected significantly less than older leaves based on measurements of tissue damage. To identify quantitative changes to transcriptomes between young and old leaves upon heat stress, we used RNA sequencing on young and old leaves from plants exposed to control and heat stress at 42 °C for 1 h and 10 h. A total of 6472 differentially expressed genes between young and old leaf were identified under control condition, and 9126 and 6891 under 1 h and 10 h heat stress, respectively. Analyses of differentially expressed transcripts led to the identification of multiple functional clusters of genes that may have potential roles in the increased heat tolerance of young leaves including higher level of expression in young leaves of genes encoding chaperones, heat shock proteins and proteins known in oxidative stress resistance. Differential levels of transcripts for genes implicated in pectin metabolism, cutin and wax biosynthesis, pentose and glucuronate interconversions, cellulose degradation, indole glucosinolate metabolism and RNA splicing between young and old leaves under heat stress suggest that cell wall remodelling, cuticular wax synthesis and carbohydrate modifications impacted by alternative splicing may also have roles in the improved heat stress tolerance of young leaves.

Advances and perspectives in the metabolomics of stomatal movement and the disease triangle.

Crops are continuously exposed to microbial pathogens that cause tremendous yield losses worldwide. Stomatal pores formed by pairs of specialized guard cells in the leaf epidermis represent a major route of pathogen entry. Guard cells have an essential role as a first line of defense against pathogens. Metabolomics is an indispensable systems biology tool that has facilitated discovery and functional studies of metabolites that regulate stomatal movement in response to pathogens and other environmental factors. Guard cells, pathogens and environmental factors constitute the "stomatal disease triangle". The aim of this review is to highlight recent advances toward understanding the stomatal disease triangle in the context of newly discovered signaling molecules, hormone crosstalk, and consequent molecular changes that integrate pathogens and environmental sensing into stomatal immune responses. Future perspectives on emerging single-cell studies, multiomics and molecular imaging in the context of stomatal defense are discussed. Advances in this important area of plant biology will inform rational crop engineering and breeding for enhanced stomatal defense without disruption of other pathways that impact crop yield.

The Arabidopsis MIK2 receptor elicits immunity by sensing a conserved signature from phytocytokines and microbes.

Sessile plants encode a large number of small peptides and cell surface-resident receptor kinases, most of which have unknown functions. Here, we report that the Arabidopsis receptor kinase MALE DISCOVERER 1-INTERACTING RECEPTOR-LIKE KINASE 2 (MIK2) recognizes the conserved signature motif of SERINE-RICH ENDOGENOUS PEPTIDEs (SCOOPs) from Brassicaceae plants as well as proteins present in fungal Fusarium spp. and bacterial Comamonadaceae, and elicits various immune responses. SCOOP signature peptides trigger immune responses and altered root development in a MIK2-dependent manner with a sub-nanomolar sensitivity. SCOOP12 directly binds to the extracellular leucine-rich repeat domain of MIK2 in vivo and in vitro, indicating that MIK2 is the receptor of SCOOP peptides. Perception of SCOOP peptides induces the association of MIK2 and the coreceptors SOMATIC EMBRYOGENESIS RECEPTOR KINASE 3 (SERK3) and SERK4 and relays the signaling through the cytosolic receptor-like kinases BOTRYTIS-INDUCED KINASE 1 (BIK1) and AVRPPHB SUSCEPTIBLE1 (PBS1)-LIKE 1 (PBL1). Our study identifies a plant receptor that bears a dual role in sensing the conserved peptide motif from phytocytokines and microbial proteins via a convergent signaling relay to ensure a robust immune response.

New aspects of iron-copper crosstalk uncovered by transcriptomic characterization of Col-0 and the copper uptake mutant spl7 in Arabidopsis thaliana.

Iron (Fe) and copper (Cu) are essential micronutrients for energy metabolism and reactive oxygen species (ROS) scavenging. Some Cu-containing proteins can be substituted with Fe-containing proteins, and vice versa, while several Arabidopsis genes are regulated by both metals. Few details of how plants coordinate Fe-Cu crosstalk are known. Gene expression was measured in the roots and rosettes of Fe, Cu, and simultaneously Fe and Cu deficient WT plants and a mutant of the Cu-uptake transcription factor SPL7. The spl7 mutant accumulated excess Fe under normal conditions, and lower Fe supply rescued the growth phenotype and normalized the Fe : Cu ratios. Most Fe regulated genes were expressed similarly in the WT and spl7 mutant, although at higher fold-change levels in spl7 mutants. Expression patterns indicated that both SPL7 and the FIT Fe uptake transcription factor influenced the expression of many key Fe uptake genes. Most notably, the newly discovered IMA/FEP genes and the subgroup Ib bHLH genes, which are upstream of Fe uptake responses, were repressed in the WT under Cu deficiency. Several AP2/ethylene response factor (AP2/ERF) genes and other redox homeostasis network genes were derepressed in spl7 mutants. Together, we present new information about Fe-Cu crosstalk in plants that could be applied for developing abiotic stress tolerant crops.