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Bleeker, P.

Publications and source records attributed to Bleeker, P..

4 recordsLinked to original sources

Modeling of Glucosinolate Biosynthesis During Biotic Stress as a Function of mRNA

Glucosinolates are an important group of specialized metabolites in the Brassicaceae family, playing a role as defensive compounds against biotic attackers. In response to biotic stress, plants upregulate glucosinolate biosynthesis in part by increasing the abundance of enzymes in the glucosinolate biosynthetic pathway. As an increase in enzyme abundance is often preceded by an increase in the corresponding mRNA levels, the dynamic changes in mRNA levels should capture the information required to infer how metabolite levels change over time. In order to test this hypothesis, a time series of experimental glucosinolate content data collected from Arabidopsis thaliana, exposed to either a mock or methyl jasmonate (MeJA) treatment, as a proxy for biotic stress, was combined with existing mRNA abundance data over time at the same developmental stage and treatment. We propose the GEEM model, a multilevel mechanistic ordinary differential equation (ODE) model, which goes from Gene expression to an enzyme level model, followed by a Michaelis Menten kinetics metabolite model, to simulate the dynamics of a segment of the indolic glucosinolate pathway. In order to constrain the GEEM model, three models were fit to experimental de novo specialized metabolite data, using different degrees of freedom by utilizing both a Gradient Boosted Tree model with a tested architecture to predict the kinetic constants, and augmenting these predictions with a literature review of the known Michaelis Menten kinetic constants from the glucosinolate pathway. Using Sequential Monte Carlo - Approximate Bayesian Computing to fit the GEEM model to the experimental data, we showed that given the mRNA levels and initial concentrations of metabolites, the changes in specialized metabolites over time and treatment can be modeled. Author SummaryWe study how plants adjust their natural chemical defenses over time when they are under attack from living organisms. In the mustard family, including the subject of our experiment Arabidopsis, one important group of defense chemicals is called glucosinolates. When Arabidopsis is under attack, certain gene pathways can be activated or deactivated, allowing the plant to modulate the amount of enzymes they produce, which in turn modulates the levels of these defensive chemicals. In this work, we combine measurements of gene activity and glucosinolate levels from Arabidopsis treated with a compound used in stress signal that mimics insect or pathogen attack. We then constructed a mathematical model that goes from gene activity, to amount of enzyme present, and ends with the amounts of specific glucosinolates over time. By fitting this model to experimental data, we show that it is possible to predict how glucosinolate levels change over time from the gene activity and initial glucosinolate levels. Our approach offers a way to connect gene expression datasets to real changes in plant defense chemistry, with potential applications in plant breeding and insight into how these pathways change due to stress.

systems biology↗

Glandular Trichome Rupture in Tomato Plants is an Ultra-Fast & Sensitive Defense Mechanism Against Insects

Trichomes, specialized hair-like structures on the surfaces of many plants, play a crucial role in defense against herbivorous insects. We investigated the biomechanics of type VI glandular trichomes in cultivated tomato (Solanum lycopersicum) and its wild relative (Solanum habrochaites). Using micropipette force sensors and high-speed imaging, we uncovered the rupture mechanics underlying gland bursting, highlighting the small forces and short time-scales involved in this process. Additionally, we observed larvae of the Western flower thrips (Frankliniella occidentalis), a major pest in tomato cultivation, inadvertently triggering trichome rupture and accumulating glandular secretions on their bodies. These findings demonstrate how rapid gland bursting and the fluid dynamics of glandular secretions act as an efficient and swift plant defense mechanism against insect herbivory.

biophysics↗

Woolly mutation with Get02 locus overcomes the polygenic nature of trichome-based pest resistance in tomato

Type-IV glandular trichomes, which only occur in the juvenile phase of tomato development, produce acylsugars (AS) that broadly protect against arthropod herbivory. Previously, we introgressed the capacity to retain type-IV trichomes in the adult phase from Solanum galapagense into the cv. Micro-Tom (MT). The resulting MT-Get line contained five loci associated with enhancing the density of type-IV trichomes in adult plants. We genetically dissected MT-Get and obtained a sub-line containing only the locus on chromosome 2 (MT- Get02). This genotype displayed about half the density of type-IV trichomes compared to the wild progenitor. However, when we stacked the gain-of-function allele of WOOLLY, which codes for a HD-ZIP IV transcription factor, MT-Get02/Wo exhibited double the number of type-IV trichomes compared to S. galapagense. This discovery corroborates previous reports positioning WOOLLY as a master regulator of trichome development. AS levels in MT-Get02/Wo were comparable to the wild progenitor, although the composition of AS types differed, especially regarding less AS with medium-length acyl chains. Agronomical parameters of MT-Get02/Wo, including yield, were comparable to MT. Pest resistance assays showed enhanced protection against whitefly, caterpillar, and the fungus Septoria lycopersici. However, resistance levels did not reach that of the wild progenitor, suggesting the specificity of particulars AS types in the pest resistance mechanism. Our findings in trichome-mediated resistance advance the development of robust, naturally resistant tomato varieties, harnessing the potential of natural genetic variation. Moreover, by manipulating only two loci, we achieved exceptional results for a highly complex, polygenic trait, such as herbivory resistance in tomato.

plant biology↗

Genetic factors outside the metabolic cluster for plastid-derived sesquiterpenes are required to pursuit arthropod-resistant tomatoes

To deal with arthropod pests the tomato wild relatives produce a variety of defense compounds in their glandular trichomes. In Solanum habrochaites LA1777, a functional cluster of genes on chromosome 8 controls plastid-derived sesquiterpene synthesis not found in cultivated tomatoes. The main genes at the cluster are Z-prenyltransferase (zFPS) that produces Z-Z-farnesyl diphosphate (Z,Z-FPP), and Santalene and Bergamotene Synthase (SBS) that uses Z,Z-FPP to produce -santalene, {beta}-bergamotene, and -bergamotene in type-VI glandular trichomes. Both LA1777 and cultivated tomatoes have type-VI trichomes, but the gland in cultivated tomato is much smaller containing low levels of monoterpenes and cytosolic-derived sesquiterpenes, which do not provide tomato with the same pest resistance as in LA1777. We successfully transferred the plastid-derived sesquiterpene pathway from LA1777 to type-VI trichomes of a cultivated tomato (cv. Micro-Tom, MT) by a back-crossing approach. The trichomes of the introgressed line named MT-Sesquiterpene synthase 2 (MT-Sst2) produced even higher levels of -santalene, {beta}-bergamotene, and -bergamotene than the type-VI glandular trichomes of LA1777. We also noticed that the type-VI trichome internal storage-cavity size increases in MT-Sst2, probably as an "inflated balloon" effect of the increased amount of sesquiterpenes. Surprisingly, the presence of high amounts of plastid-derived sesquiterpenes was not sufficient to confer resistance to various tomato pests in MT-Sst2. Since MT-Sst2 made the same sesquiterpenes as LA1777, this points to additional factors, outside the genomic region thought to be the metabolic cluster, necessary to obtain arthropod-resistant tomatoes. Our results also provide a better understanding of the morphology of S. habrochaites type-VI trichomes. One-sentence summaryCultivated tomatoes harboring the plastid-derived sesquiterpenes from S. habrochaites need additional genetic components necessary to convert them into effective insecticides.

plant biology↗