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Biomedical subjects

Thomas Mitchell-Olds

Publications and source records attributed to Thomas Mitchell-Olds.

11 recordsLinked to original sources

Evolutionary and ecological functional genomics.

A unique combination of disciplines is emerging--evolutionary and ecological functional genomics--which focuses on the genes that affect ecological success and evolutionary fitness in natural environments and populations. Already this approach has provided new insights that were not available from its disciplinary components in isolation. However, future advances will necessitate the re-engineering of scientific attitudes, training and institutions, to achieve extensive multidisciplinarity.

Animals↗

Multiple hybrid formation in natural populations: concerted evolution of the internal transcribed spacer of nuclear ribosomal DNA (ITS) in North American Arabis divaricarpa (Brassicaceae).

DNA sequence variation of the internal transcribed spacer (ITS) region of nuclear ribosomal DNA from Arabis holboellii, A. drummondii, and its putative hybrid A. divaricarpa was analyzed to study hybrid speciation in a species system geographically covering nearly the entire North American continent. Based on molecular systematics the investigated species are better combined under the genus Boechera. Multiple intraindividual ITS copies were detected in numerous accessions of A. divaricarpa, and, to a minor extent, in the parental taxa. Comparative phylogenetic analysis demonstrates that reticulate evolution is common. Consequently, concerted evolution of ITS regions resulted in different types of ITS fragments not only in hybrid populations but also in one of the parental taxa, A. holboellii. Hybrid formation often occurred independently at different sites and at different times, which is reflected by ITS copies resampling the original parental sequence variation in different ways. Some biogeographic structuring of genetic diversity is apparent and mirrors postglacial migration routes. Hybridization, reticulation, and apomixis are assumed to be the major forces driving speciation processes in this species complex. Analysis of conserved regions and secondary structures of the ITS region provided no evidence that, in this system, hybrid ITS evolution is predominantly driven in a particular direction. However, two regions in the ITS1 and ITS2, respectively, show higher mutation rates than expected from outgroup comparisons. Strong evidence for the occurrence of apomixis in A. holboellii and A. divaricarpa has come from pollen size measurements and estimations of pollen quality, which favor the hypothesis that A. drummondii served as paternal hybridization partner more frequently than A. holboellii.

Arabis↗

Disarming the mustard oil bomb.

Plants are attacked by a broad array of herbivores and pathogens. In response, plants deploy an arsenal of defensive traits. In Brassicaceae, the glucosinolate-myrosinase complex is a sophisticated two-component system to ward off opponents. However, this so-called "mustard oil bomb" is disarmed by a glucosinolate sulfatase of a crucifer specialist insect, diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae). Sulfatase activity of this enzyme largely prevents the formation of toxic hydrolysis products arising from this plant defense system. Importantly, the enzyme acts on all major classes of glucosinolates, thus enabling diamondback moths to use a broad range of cruciferous host plants.

Amino Acid Sequence↗

Evidence for regulation of resistance in Arabidopsis to Egyptian cotton worm by salicylic and jasmonic acid signaling pathways.

Signaling cross-talk between wound- and pathogen-response pathways influences resistance of plants to insects and disease. To elucidate potential interactions between salicylic acid (SA) and jasmonic acid (JA) defense pathways, we exploited the availability of characterized mutants of Arabidopsis thaliana (L.) Heynh. and monitored resistance to Egyptian cotton worm (Spodoptera littoralis Boisd.; Lepidoptera: Noctuidae). This generalist herbivore is sensitive to induced plant defense pathways and is thus a useful model for a mechanistic analysis of insect resistance. As expected, treatment of wild-type Arabidopsis with JA enhanced resistance to Egyptian cotton worm. Conversely, the coil mutant, with a deficiency in the JA response pathway, was more susceptible to Egyptian cotton worm than wild-type Arabidopsis. By contrast, the nprl mutant, with defects in systemic disease resistance, exhibited enhanced resistance to Egyptian cotton worm. Pretreatment with SA significantly reduced this enhanced resistance of nprl plants but had no influence on the resistance of wild-type plants. However, exogenous SA reduced the amount of JA that Egyptian cotton worm induced in both npr1 mutant and wild-type plants. Thus, this generalist herbivore engages two different induced defense pathways that interact to mediate resistance in Arabidopsis.

Animals↗

Benzoic acid glucosinolate esters and other glucosinolates from Arabidopsis thaliana.

The spectacular recent progress in Arabidopsis thaliana molecular genetics furnishes outstanding tools for studying the formation and function of all metabolites in this cruciferous species. One of the major groups of secondary metabolites in A. thaliana is the glucosinolates. These hydrophilic, sulfur-rich glycosides appear to serve as defenses against some generalist herbivores and pathogens, and as feeding and oviposition stimulants to specialist herbivores. To help study their biosynthesis and role in plant-insect interactions, we wanted to determine the complete glucosinolate content of A. thaliana. In previous studies, 24 glucosinolates had been identified from ecotype Columbia. We reinvestigated Columbia as well as additional ecotypes and mutant lines, and identified 12 further glucosinolates, including five novel compounds. Structures were elucidated by MS and NMR spectroscopy of their desulfated derivatives, and by enzymatic cleavage of the attached ester moieties. Four of the novel glucosinolates are benzoate esters isolated from the seeds. In all but one of these compounds, esterification is on the glucose moiety rather than the side chain, a very unusual feature for glucosinolates. Among additional glucosinolates identified were the first non-chain elongated, methionine-derived glucosinolate from A. thaliana and the first compounds that appear to be derived from leucine.

Arabidopsis↗

Plant evolutionary genomics.

Evolutionary genomics combines functional and evolutionary analyses of genome conservation and differentiation. Gene duplication and polyploidy have fundamentally shaped the genomes of Arabidopsis and all angiosperms. Recent comparative studies have focussed on gene regulation, the function of untranscribed genomic regions, and the effects of natural selection on protein function. A large fraction of interspecific protein divergence is probably adaptive, and may be useful for experimental studies of genes and proteins.

Animals↗

Local and differential control of vegetative storage protein expression in response to herbivore damage in Arabidopsis thaliana.

Vegetative storage proteins (VSPs) are thought to fulfil important nutritional roles during plant development and stress adaptation. Plant responses to mechanical wounding and herbivore damage include an activation of VSP expression. It was recently suggested that vsp is part of the systemic response of Arabidopsis to wounding. To test this proposal, we monitored the spatial regulation of vsp mRNAs and VSP proteins. Arabidopsis contains two vsp genes and real-time quantitative PCR allowed us to characterize their differential expression. The ratio of vsp1 to vsp2 mRNA abundance increased when plants were challenged with diamondback moth larvae or Egyptian cotton worms, but not when they were mechanically wounded. We observed a dramatic increase of vsp1 and vsp2 mRNA as well as VSP protein levels in leaves that experienced herbivore damage. By contrast, there was a relatively minor increase of vsp mRNA and VSP protein levels in undamaged leaves of infested plants. These results clearly demonstrate that VSPs are part of the local plant response to herbivore attack. To obtain additional information on vsp regulation, we analysed a fusion of a soybean vspB promoter fragment to the beta-glucuronidase gene in transgenic Arabidopsis plants. The vspB promoter responded to both jasmonate and herbivore treatments, suggesting that similar signals regulate its expression in both plant species.

Journal Article↗

Comparative analysis of quantitative trait loci controlling glucosinolates, myrosinase and insect resistance in Arabidopsis thaliana.

Evolutionary interactions among insect herbivores and plant chemical defenses have generated systems where plant compounds have opposing fitness consequences for host plants, depending on attack by various insect herbivores. This interplay complicates understanding of fitness costs and benefits of plant chemical defenses. We are studying the role of the glucosinolate-myrosinase chemical defense system in protecting Arabidopsis thaliana from specialist and generalist insect herbivory. We used two Arabidopsis recombinant inbred populations in which we had previously mapped QTL controlling variation in the glucosinolate-myrosinase system. In this study we mapped QTL controlling resistance to specialist (Plutella xylostella) and generalist (Trichoplusia ni) herbivores. We identified a number of QTL that are specific to one herbivore or the other, as well as a single QTL that controls resistance to both insects. Comparison of QTL for herbivory, glucosinolates, and myrosinase showed that T. ni herbivory is strongly deterred by higher glucosinolate levels, faster breakdown rates, and specific chemical structures. In contrast, P. xylostella herbivory is uncorrelated with variation in the glucosinolate-myrosinase system. This agrees with evolutionary theory stating that specialist insects may overcome host plant chemical defenses, whereas generalists will be sensitive to these same defenses.

Animals↗

Recombination and gene conversion in a 170-kb genomic region of Arabidopsis thaliana.

Arabidopsis thaliana is a highly selfing plant that nevertheless appears to undergo substantial recombination. To reconcile its selfing habit with the observations of recombination, we have sampled the genetic diversity of A. thaliana at 14 loci of approximately 500 bp each, spread across 170 kb of genomic sequence centered on a QTL for resistance to herbivory. A total of 170 of the 6321 nucleotides surveyed were polymorphic, with 169 being biallelic. The mean silent genetic diversity (pi(s)) varied between 0.001 and 0.03. Pairwise linkage disequilibria between the polymorphisms were negatively correlated with distance, although this effect vanished when only pairs of polymorphisms with four haplotypes were included in the analysis. The absence of a consistent negative correlation between distance and linkage disequilibrium indicated that gene conversion might have played an important role in distributing genetic diversity throughout the region. We tested this by coalescent simulations and estimate that up to 90% of recombination is due to gene conversion.

Arabidopsis↗

Genetic architecture of plastic methyl jasmonate responses in Arabidopsis thaliana.

The ability of a single genotype to generate different phenotypes in disparate environments is termed phenotypic plasticity, which reflects the interaction of genotype and environment on developmental processes. However, there is controversy over the definition of plasticity genes. The gene regulation model states that plasticity loci influence trait changes between environments without altering the means within a given environment. Alternatively, the allelic sensitivity model argues that plasticity evolves due to selection of phenotypic values expressed within particular environments; hence plasticity must be controlled by loci expressed within these environments. To identify genetic loci controlling phenotypic plasticity and address this controversy, we analyzed the plasticity of glucosinolate accumulation under methyl jasmonate (MeJa) treatment in Arabidopsis thaliana. We found genetic variation influencing multiple MeJa signal transduction pathways. Analysis of MeJa responses in the Landsberg erecta x Columbia recombinant inbred lines identified a number of quantitative trait loci (QTL) that regulate plastic MeJa responses. All significant plasticity QTL also impacted the mean trait value in at least one of the two "control" or "MeJa" environments, supporting the allelic sensitivity model. Additionally, we present an analysis of MeJa and salicylic acid cross-talk in glucosinolate regulation and describe the implications for glucosinolate physiology and functional understanding of Arabidopsis MeJa signal transduction.

Acetates↗