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Highlights • Strategies for future high throughput, non-destructive and cost-efficient measurement of plant traits are highlighted. • Use of low-cost and DIY approaches in phenomics provides opportunities for rapid prototyping and sensor development. • Robust protocols, data harmonization and provenance are critical to allow data reuse and cross validation of phenotypes. • Below-ground phenotyping is a major bottleneck and new technologies allowing the measurement of root-related traits are needed. Abstract At the 4th International Plant Phenotyping Symposium meeting of the International Plant Phenotyping Network (IPPN) in 2016 at CIMMYT in Mexico, a workshop was convened to consider ways forward with sensors for phenotyping. The increasing number of field applications provides new challenges and requires specialised solutions. There are many traits vital to plant growth and development that demand phenotyping approaches that are still at early stages of development or elude current capabilities. Further, there is growing interest in low-cost sensor solutions, and mobile platforms that can be transported to the experiments, rather than the experiment coming to the platform. Various types of sensors are required to address diverse needs with respect to targets, precision and ease of operation and readout. Converting data into knowledge, and ensuring that those data (and the appropriate metadata) are stored in such a way that they will be sensible and available to others now and for future analysis is also vital. Here we are proposing mechanisms for "next generation phenomics" based on our learning in the past decade, current practice and discussions at the IPPN Symposium, to encourage further thinking and collaboration by plant scientists, physicists and engineering experts. [ABSTRACT FROM AUTHOR]

Summary: Very few CACTA transposon-like sequences have been described in Solanaceae species. Sequence information has been restricted to partial transposase (TPase)-like fragments, and no target gene of CACTA-like transposon insertion has been described in tomato to date. In this manuscript, we report on a CACTA transposon-like insertion in intron I of tomato (Lycopersicon esculentum) invertase gene Lin5 and TPase-like sequences of several Solanaceae species. Consensus primers deduced from the TPase region of the tomato CACTA transposon-like element allowed the amplification of similar sequences from various Solanaceae species of different subfamilies including Solaneae (Solanum tuberosum), Cestreae (Nicotiana tabacum) and Datureae (Datura stramonium). This demonstrates the ubiquitous presence of CACTA-like elements in Solanaceae genomes. The obtained partial sequences are highly conserved, and allow further detection and detailed analysis of CACTA-like transposons throughout Solanaceae species. CACTA-like transposon sequences make possible the evaluation of their use for genome analysis, functional studies of genes and the evolutionary relationships between plant species. [Copyright &y& Elsevier]

The disaccharide sucrose and the cleavage products glucose and fructose are the central molecules for carbohydrate translocation, metabolism and sensing in higher plants. Invertases mediate the hydrolytic cleavage of sucrose into the hexose monomers. Plants possess three types of invertases, which are located in the apoplast, the cytoplasm and the vacuole, respectively. It has become evident that extracellular and vacuolar invertase isoenzymes are key metabolic enzymes that are involved in various aspects of the plant life cycle and the response of the plant to environmental stimuli because their substrates and reaction products are both nutrients and signal molecules. Invertases, alone or in combination with plant hormones, can regulate many aspects of the growth and development of plants from gene expression to long-distance nutrient allocation and are involved in regulating carbohydrate partitioning, developmental processes, hormone responses and biotic and abiotic interactions. [Copyright &y& Elsevier]

Seed inoculation with entomopathogenic fungi (EPF) causes plant-mediated effects against arthropod herbivores, but the responses vary among EPF isolates. We used a wheat model system with three isolates representing Beauveria bassiana and Metarhizium spp. causing either negative or positive effects against the aphid Rhopalosiphum padi. Activities of six carbohydrate enzymes increased in plants showing biomass build-up after EPF inoculations. However, only aldolase activity showed positive correlation with R. padi numbers. Plants inoculated with M. robertsii hosted fewest aphids and showed increased activity of superoxide dismutase, implying a defense strategy of resistance towards herbivores. In M. brunneum -inoculated plants, hosting most R. padi , activities of catalase and glutathione reductase were increased suggesting enhanced detoxification responses towards aphids. However, M. brunneum simultaneously increased plant growth indicating that this isolate may cause the plant to tolerate herbivory. EPF seed inoculants may therefore mediate either tolerance or resistance towards biotic stress in plants in an isolate-dependent manner. [ABSTRACT FROM AUTHOR]

Abstract: Phytoalexins are pathogen induced low molecular weight compounds with antimicrobial activities derived from secondary metabolism. Following their identification, phytoalexins were directly incorporated into the network of plant defense responses. Due to their heterogeneity, the metabolic pathways involved in phytoalexin formation and in particular the regulatory mechanisms remained elusive. Consequently, research focus shifted to the characterization of other components of plant immunity such as defense signaling and resistance mechanisms, including components of systemic acquired and induced systemic resistance, effector and pathogen-associated molecular pattern triggered immunity as well as R-gene resistance. Despite the obtained knowledge on these immunity mechanisms, genetic engineering employing these mechanisms and classical breeding reached too low improvements in crop protection, probably because classical breeding focused on yield performance and taste, rather than pathogen resistance. The increasing demand for disease resistant crop species and the aim to reduce pesticide application therefore requires alternative approaches. Recent advances in the understanding of phytoalexin function, biosynthesis and regulation, in combination with novel methods of molecular engineering and advances in instrumental analysis, returned attention to phytoalexins as a potent target for improving crop protection. Based on this, the advantages as well as potential bottlenecks for molecular approaches of modulating inducible phytoalexins to improve crop protection are discussed. [Copyright &y& Elsevier]

Plant growth-promoting and stress resilience-inducing root endophytic fungi represent an additional carbohydrate sink. This study aims to test if such root endophytes affect the sugar metabolism of the host plant to divert the flow of resources for their purposes. Fresh and dry weights of roots and shoots of tomato (Solanum lycopersicum) colonised by the closely related Serendipita indica and Serendipita herbamans were recorded. Plant carbohydrate metabolism was analysed by measuring sugar levels, by determining activity signatures of key enzymes of carbohydrate metabolism, and by quantifying mRNA levels of genes involved in sugar transport and turnover. During the interaction with the tomato plants, both fungi promoted root growth and shifted shoot biomass from stem to leaf tissues, resulting in increased leaf size. A common effect induced by both fungi was the inhibition of phosphofructokinase (PFK) in roots and leaves. This glycolytic-pacing enzyme shows how the glycolysis rate is reduced in plants and, eventually, how sugars are allocated to different tissues. Sucrose phosphate synthase (SPS) activity was strongly induced in colonised roots. This was accompanied by increased SPS-A1 gene expression in S. herbamans -colonised roots and by increased sucrose amounts in roots colonised by S. indica. Other enzyme activities were barely affected by S. indica , but mainly induced in leaves of S. herbamans -colonised plants and decreased in roots. This study suggests that two closely related root endophytic fungi differentially influence plant carbohydrate metabolism locally and systemically, but both induce a similar increase in plant biomass. Notably, both fungal endophytes induce an increase in SPS activity and, in the case of S. indica, sucrose resynthesis in roots. In leaves of S. indica -colonised plants, SWEET11b expression was enhanced, thus we assume that excess sucrose was exported by this transporter to the roots. ‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬ [ABSTRACT FROM AUTHOR]

Plant growth-promoting rhizobacteria (PGPR) are known for exerting beneficial effects on plant growth and tolerance to plant pathogens. However, their specific role in mediating protection against abiotic stress remains underexplored. The aim of this study was to characterise the ability of the cytokinin-producing beneficial bacterium Pseudomonas fluorescens G20-18 to enhance tomato growth and boost tolerance to drought stress. Tomato seedlings were root inoculated and their growth and physiological and molecular responses assessed under well-watered conditions and also in response to progressive drought stress and a subsequent recovery period. Root inoculation with G20-18 had a significant positive impact on tomato growth. Furthermore, G20-18 inoculated and drought-stressed plants showed higher leaf chlorophyll and abscisic acid (ABA) content and stomatal closure than non-inoculated controls. Root inoculation also increased the activity of different carbohydrate metabolism enzymes, which are important for root and leaf growth and development in drought stressed plants. A significant increase in the activity of different antioxidant enzymes and total antioxidant capacity correlated with elevated levels of relevant secondary metabolites, such as phenolics, anthocyanins and flavonoids. RNA sequencing revealed distinct qualitative and quantitative differences in gene regulation in response to G20-18. Notably, the number of genes differentially regulated in response to G20-18 was approximately sevenfold higher during drought stress, indicating that root inoculation with the bacteria primed the plants for a much stronger transcriptionally regulated systemic drought stress response. The regulated genes are related to phenylalanine metabolism and other key processes linked to plant growth, development and drought stress resilience. A role of the ability of G20-18 to produce the plant hormone cytokinin for interaction with tomato was established by the cytokinin-deficient biosynthesis mutants CNT1 and CNT2. In comparison with G20-18, the inoculation of plants with CNT1 resulted in a reduced number of differentially regulated genes. The relative change was most prominent under well-watered conditions with a 85 % reduction, corresponding to 462 genes. However, under drought conditions the absolute number of differentially regulated genes was reduced by even 2219 in response to the CNT1 mutant. The relevance of the ability of G20-18 to produce cytokinins for interaction with plants was also evident from differences in growth and specific cell and ecophysiological parameters in response to CNT1 and CNT2. These findings provide novel insights about G20-18's ability to improve drought stress responses and the role of interkingdom signalling by bacterial-derived cytokinins, and contribute to enhance the robustness of the practical application of these microorganisms to improve crop resilience in agricultural production. [ABSTRACT FROM AUTHOR]

[Display omitted] • Phenomics approach to assess infection of four barley genotypes by P. teres. • Identification of an enzyme activity fingerprint for resistance against P. teres. • Distinct temporal dynamics of gene expression profiles reflect pathogen resistance. • Resistance correlates with a specific reflectance and fluorescence signature. • Pre-symptomatic detection of pathogen infection by sensor-based imaging. Necrotic and chlorotic symptoms induced during Pyrenophora teres infection in barley leaves indicate a compatible interaction that allows the hemi-biotrophic fungus Pyrenophora teres to colonise the host. However, it is unexplored how this fungus affects the physiological responses of resistant and susceptible cultivars during infection. To assess the degree of resistance in four different cultivars, we quantified visible symptoms and fungal DNA and performed expression analyses of genes involved in plant defence and ROS scavenging. To obtain insight into the interaction between fungus and host, we determined the activity of 19 key enzymes of carbohydrate and antioxidant metabolism. The pathogen impact was also phenotyped non-invasively by sensor-based multireflectance and –fluorescence imaging. Symptoms, regulation of stress-related genes and pathogen DNA content distinguished the cultivar Guld as being resistant. Severity of net blotch symptoms was also strongly correlated with the dynamics of enzyme activities already within the first day of infection. In contrast to the resistant cultivar, the three susceptible cultivars showed a higher reflectance over seven spectral bands and higher fluorescence intensities at specific excitation wavelengths. The combination of semi high-throughput physiological and molecular analyses with non-invasive phenotyping enabled the identification of bio-signatures that discriminates the resistant from susceptible cultivars. [ABSTRACT FROM AUTHOR]

Abstract: The allene oxide cyclase (AOC) catalyzes the formation of cis-(+)-12-oxophytodienoic acid, an intermediate in jasmonate biosynthesis and is encoded by a single copy gene in tomato. The full length AOC promoter isolated by genome walk contains 3600bp. Transgenic tomato lines carrying a 1000bp promoter fragment and the full length promoter, respectively, in front of the β-glucuronidase (GUS)-encoding uidA gene and several tobacco lines carrying the full length tomato AOC promoter before GUS were used to record organ- and tissue-specific promoter activities during development and in response to various stimuli. High promoter activities corresponding to immunocytochemically detected occurrence of the AOC protein were found in seeds and young seedlings and were confined to the root tip, hypocotyl and cotyledons of 3-d-old seedlings. In 10-d-old seedlings promoter activity appeared preferentially in the elongation zone. Fully developed tomato leaves were free of AOC promoter activity, but showed high activity upon wounding locally and systemically or upon treatment with JA, systemin or glucose. Tomato flowers showed high AOC promoter activities in ovules, sepals, anthers and pollen. Most of the promoter activity patterns found in tomato with the 1000bp promoter fragment were also detected with the full length tomato AOC promoter in tobacco during development or in response to various stimuli. The data support a spatial and temporal regulation of JA biosynthesis during development and in response to environmental stimuli. [Copyright &y& Elsevier]

Abstract: The initiation of defence responses in plants is accompanied by fundamental changes in gene expression: the expression of pathogenesis-related genes is co-ordinately regulated with metabolic changes such as down regulation of photosynthesis and induction of sink metabolism. To identify candidate regulators of this co-ordinated regulatory mechanism, the role of WRKY transcription factors in the initiation of defence response was analysed in tomato. A WRKY-type transcription factor (LpWRKY1) from tomato was cloned by a reverse Northern approach. The corresponding mRNA is rapidly and transiently induced after challenging the cells with an elicitor-preparation derived from the wilt inducing fungus Fusarium oxysporum lycopersici (E-FOL) and the fungal elicitor chitosan, whereas the endogenous signals systemin and salicylic acid are inactive. Inhibition of protein biosynthesis by cycloheximide results in sustained induction of mRNA for LpWRKY1. In contrast, the transient induction of the gene encoding LpWRKY1 in response to elicitation by E-FOL is inhibited by the protein-kinase inhibitor staurosporine and may be mimicked by the phosphatase inhibitors endothall and cantharidine indicating the involvement of protein phosphorylation in the regulation of WRKY-type transcription factors. Direct proof of this postranslational modification of LpWRKY1 was obtained by demonstrating in-gel kinase assays using recombinant LpWRKY1 as substrate. A 44kDa and a 67kDa protein kinase were shown to be transiently activated to phosphorylate LpWRKY1 protein in response to elicitation with E-FOL. [Copyright &y& Elsevier]

This study was conducted to understand the mechanism of wheat yield decrease under drought stress and the role of CO 2 in modulating physiological and metabolic drought effects. Wheat was grown under ambient and elevated CO 2 (400 and 800 ppm, respectively), and plants were subjected to drought stress prior to anthesis. Photosynthetic rate (An), stomatal conductance (Gs), transpiration rate (E) and activities of carbohydrate metabolic enzymes were decreased in leaf and increased in spikes during drought. Total antioxidant potential (TAP) was decreased under drought both in leaf and spike. Grain yield parameters were again reduced under drought, while An, E and most of the yield traits were increased under elevated CO 2. The number of grains spike-1 correlated positively with An, TAP and cell wall invertase activity, while it negatively correlated with ascorbate peroxidase, cell wall peroxidase and glutathione reductase activities in leaves. Thousand kernel weight positively correlated with leaf phosphoglucoisomerase and spike glucose-6-phosphate dehydrogenase activities. This indicates that elevated CO 2 could boost CO 2 assimilation through an increase in antioxidant potential and facilitate more photosynthate supply via various increased carbohydrate metabolic enzyme activities, and thus increases yield. This could be a possible mechanism of grain yield increase caused by elevated CO 2. • Drought reduces grain yield via reduction in antioxidants potential, photosynthetic rate, and carbohydrate-metabolic enzymes. • Elevated CO 2 can boost CO 2 assimilation via improved antioxidants activity, WUE, and carbohydrate-metabolic enzyme activities. • Elevated CO 2 ameliorates the adverse impact of drought through enhanced antioxidant potential, WUE, and source-sink balance. [ABSTRACT FROM AUTHOR]