Decoding the sorghum methylome: understanding epigenetic contributions to agronomic traits

DNA methylation is a chromatin modification that plays an essential role in regulatinggene expression and genome stability and it is typically associated with gene silencingand heterochromatin. Owing to its heritability, alterations in the patterns of DNA methyla-tion have the potential to provide for epigenetic inheritance of traits. Contemporary epige-nomic technologies provide information beyond sequence variation and could supplyalternative sources of trait variation for improvement in crops such as sorghum. Yet, com-pared with other species such as maize and rice, the sorghum DNA methylome is farless well understood. The distribution of CG, CHG, and CHH methylation in the genomeis different compared with other species. CG and CHG methylation levels peak aroundcentromeric segments in the sorghum genome and are far more depleted in the genedense chromosome arms. The genes regulating DNA methylation in sorghum are also yetto be functionally characterised; better understanding of their identity and functional ana-lysis of DNA methylation machinery mutants in diverse genotypes will be important tobetter characterise the sorghum methylome. Here, we catalogue homologous genesencoding methylation regulatory enzymes in sorghum based on genes inArabidopsis,maize, and rice. Discovering variation in the methylome may uncover epialleles thatprovide extra information to explain trait variation and has the potential to be applied inepigenome-wide association studies or genomic prediction. DNA methylation can alsoimprove genome annotations and discover regulatory elements underlying traits. Thus,improving our knowledge of the sorghum methylome can enhance our understanding ofthe molecular basis of traits and may be useful to improve sorghum performance

Decoding the sorghum methylome: understanding epigenetic contributions to agronomic traits

DNA methylation is a chromatin modification that plays an essential role in regulatinggene expression and genome stability and it is typically associated with gene silencingand heterochromatin. Owing to its heritability, alterations in the patterns of DNA methyla-tion have the potential to provide for epigenetic inheritance of traits. Contemporary epige-nomic technologies provide information beyond sequence variation and could supplyalternative sources of trait variation for improvement in crops such as sorghum. Yet, com-pared with other species such as maize and rice, the sorghum DNA methylome is farless well understood. The distribution of CG, CHG, and CHH methylation in the genomeis different compared with other species. CG and CHG methylation levels peak aroundcentromeric segments in the sorghum genome and are far more depleted in the genedense chromosome arms. The genes regulating DNA methylation in sorghum are also yetto be functionally characterised; better understanding of their identity and functional ana-lysis of DNA methylation machinery mutants in diverse genotypes will be important tobetter characterise the sorghum methylome. Here, we catalogue homologous genesencoding methylation regulatory enzymes in sorghum based on genes inArabidopsis,maize, and rice. Discovering variation in the methylome may uncover epialleles thatprovide extra information to explain trait variation and has the potential to be applied inepigenome-wide association studies or genomic prediction. DNA methylation can alsoimprove genome annotations and discover regulatory elements underlying traits. Thus,improving our knowledge of the sorghum methylome can enhance our understanding ofthe molecular basis of traits and may be useful to improve sorghum performance

Subset of Heat-Shock Transcription Factors Required for the Early Response of Arabidopsis to Excess Light

Sunlight provides energy for photosynthesis and is essential for nearly all life on earth. However, too much or too little light or rapidly fluctuating light conditions cause stress to plants. Rapid changes in the amount of light are perceived as a change in the reduced/oxidized (redox) state of photosynthetic electron transport components in chloroplasts. However, how this generates a signal that is relayed to changes in nuclear gene expression is not well understood. We modified redox state in the reference plant, Arabidopsis thaliana, using either excess light or low light plus the herbicide DBMIB (2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone), a well-known inhibitor of photosynthetic electron transport. Modification of redox state caused a change in expression of a common set of about 750 genes, many of which are known stress-responsive genes. Among the most highly enriched promoter elements in the induced gene set were heat-shock elements (HSEs), known motifs that change gene expression in response to high temperature in many systems. We show that HSEs from the promoter of the ASCORBATE PEROXIDASE 2 (APX2) gene were necessary and sufficient for APX2 expression in conditions of excess light, or under low light plus the herbicide. We tested APX2 expression phenotypes in overexpression and loss-of-function mutants of 15 Arabidopsis A-type heat-shock transcription factors (HSFs), and identified HSFA1D, HSFA2, and HSFA3 as key factors regulating APX2 expression in diverse stress conditions. Excess light regulates both the subcellular location of HSFA1D and its biochemical properties, making it a key early component of the excess light stress network of plants

Hybrid Decay: A Transgenerational Epigenetic Decline in Vigor and Viability Triggered in Backcross Populations of Teosinte with Maize.

In the course of generating populations of maize with teosinte chromosomal introgressions, an unusual sickly plant phenotype was noted in individuals from crosses with two teosinte accessions collected near Valle de Bravo, Mexico. The plants of these Bravo teosinte accessions appear phenotypically normal themselves and the F1 plants appear similar to typical maize × teosinte F1s. However, upon backcrossing to maize, the BC1 and subsequent generations display a number of detrimental characteristics including shorter stature, reduced seed set, and abnormal floral structures. This phenomenon is observed in all BC individuals and there is no chromosomal segment linked to the sickly plant phenotype in advanced backcross generations. Once the sickly phenotype appears in a lineage, normal plants are never again recovered by continued backcrossing to the normal maize parent. Whole-genome shotgun sequencing reveals a small number of genomic sequences, some with homology to transposable elements, that have increased in copy number in the backcross populations. Transcriptome analysis of seedlings, which do not have striking phenotypic abnormalities, identified segments of 18 maize genes that exhibit increased expression in sickly plants. A de novo assembly of transcripts present in plants exhibiting the sickly phenotype identified a set of 59 upregulated novel transcripts. These transcripts include some examples with sequence similarity to transposable elements and other sequences present in the recurrent maize parent (W22) genome as well as novel sequences not present in the W22 genome. Genome-wide profiles of gene expression, DNA methylation, and small RNAs are similar between sickly plants and normal controls, although a few upregulated transcripts and transposable elements are associated with altered small RNA or methylation profiles. This study documents hybrid incompatibility and genome instability triggered by the backcrossing of Bravo teosinte with maize. We name this phenomenon "hybrid decay" and present ideas on the mechanism that may underlie it

Pleistocene divergence of two disjunct conifers in the eastern Australian temperate zone

The eastern Australian temperate biota harbours many plants with fragmented geographic ranges distributed over 1000s of kilometres, yet the spatial genetic structure of their populations remains largely unstudied. In this study, we investigated genetic variation of the nuclear internal transcribed spacer (ITS) and chloroplast DNA sequences to disentangle the phylogeography of two widely distributed but highly fragmented eastern Australian fire-sensitive temperate conifers: Callitris oblonga (12 populations and 121 individuals) and C. rhomboidea (22 populations and 263 individuals). The three highly disjunct populations of C. oblonga all had unique chloroplast and ITS haplotypes consistent with the classification of these three populations as distinct subspecies. Molecular dating indicates that divergences of these populations occurred pre- to mid- Pleistocene (2.66 to 1.08 mya). Callitris rhomboidea showed greater diversity of chloroplast haplotypes which was strongly phylogeographically structured (Gst = 0.972), with haplotypes unique to specific geographic regions. ITS haplotype diversity was far higher than in C. oblonga with 38 haplotypes displaying high geographic structuring (Gst = 0.387) with many population-specific haplotypes. A phylogeographic break was identified between populations north and south of eastern Victoria dated at 0.43–0.47 mya. In both species, the strong genetic structuring of both chloroplast and ITS haplotypes provides evidence that their widespread ranges have resulted from long term persistence in low fire frequency refugia combined with poor dispersal. Any loss of populations due to increasing fire frequency or habitat loss is likely to result in a reduction of genetic diversity

A Novel fry1 Allele Reveals the Existence of a Mutant Phenotype Unrelated to 5′->3′ Exoribonuclease (XRN) Activities in Arabidopsis thaliana Roots

BACKGROUND Mutations in the FRY1/SAL1 Arabidopsis locus are highly pleiotropic, affecting drought tolerance, leaf shape and root growth. FRY1 encodes a nucleotide phosphatase that in vitro has inositol polyphosphate 1-phosphatase and 3',(2'),5'-bisphosphate nucleotide phosphatase activities. It is not clear which activity mediates each of the diverse biological functions of FRY1 in planta. PRINCIPAL FINDINGS A fry1 mutant was identified in a genetic screen for Arabidopsis mutants deregulated in the expression of Pi High affinity Transporter 1;4 (PHT1;4). Histological analysis revealed that, in roots, FRY1 expression was restricted to the stele and meristems. The fry1 mutant displayed an altered root architecture phenotype and an increased drought tolerance. All of the phenotypes analyzed were complemented with the AHL gene encoding a protein that converts 3'-polyadenosine 5'-phosphate (PAP) into AMP and Pi. PAP is known to inhibit exoribonucleases (XRN) in vitro. Accordingly, an xrn triple mutant with mutations in all three XRNs shared the fry1 drought tolerance and root architecture phenotypes. Interestingly these two traits were also complemented by grafting, revealing that drought tolerance was primarily conferred by the rosette and that the root architecture can be complemented by long-distance regulation derived from leaves. By contrast, PHT1 expression was not altered in xrn mutants or in grafting experiments. Thus, PHT1 up-regulation probably resulted from a local depletion of Pi in the fry1 stele. This hypothesis is supported by the identification of other genes modulated by Pi deficiency in the stele, which are found induced in a fry1 background. CONCLUSIONS/SIGNIFICANCE Our results indicate that the 3',(2'),5'-bisphosphate nucleotide phosphatase activity of FRY1 is involved in long-distance as well as local regulatory activities in roots. The local up-regulation of PHT1 genes transcription in roots likely results from local depletion of Pi and is independent of the XRNs.This work was supported by an ANR-GENOPLANT grant (RIBOROOT-ANR06 GPLA 011) and the CEA agency. Array hybridizations have been partly supported by RNG (Réseau National des Génopoles, Evry, France). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. No additional external funding received for this study

Molecular and physiological responses during thermal acclimation of leaf photosynthesis and respiration in rice

To further our understanding of how sustained changes in temperature affect the carbon economy of rice (Oryza sativa), hydroponically grown plants of the IR64 cultivar were developed at 30°C/25°C (day/night) before being shifted to 25/20°C or 40/35°C. Leaf messenger RNA and protein abundance, sugar and starch concentrations, and gas‐exchange and elongation rates were measured on preexisting leaves (PE) already developed at 30/25°C or leaves newly developed (ND) subsequent to temperature transfer. Following a shift in growth temperature, there was a transient adjustment in metabolic gene transcript abundance of PE leaves before homoeostasis was reached within 24 hr, aligning with Rdark (leaf dark respiratory CO2 release) and An (net CO2 assimilation) changes. With longer exposure, the central respiratory protein cytochrome c oxidase (COX) declined in abundance at 40/35°C. In contrast to Rdark, An was maintained across the three growth temperatures in ND leaves. Soluble sugars did not differ significantly with growth temperature, and growth was fastest with extended exposure at 40/35°C. The results highlight that acclimation of photosynthesis and respiration is asynchronous in rice, with heat‐acclimated plants exhibiting a striking ability to maintain net carbon gain and growth when exposed to heat‐wave temperatures, even while reducing investment in energy‐conserving respiratory pathways.Peer reviewe

Closed curves and geodesics with two self-intersections on the Punctured torus

We classify the free homotopy classes of closed curves with minimal self intersection number two on a once punctured torus, T , up to homeomorphism. Of these, there are six primitive classes and two imprimitive. The classification leads to the topological result that, up to homeomorphism, there is a unique curve in each class realizing the minimum self intersection number. The classification yields a complete classification of geodesics on hyperbolic T which have self intersection number two. We also derive new results on the Markoff spectrum of diophantine approximation; in particular, exactly three of the imprimitive classes correspond to families of Markoff values below Hall's ray.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/41628/1/605_2005_Article_BF01317313.pd

An argument for the use of Aristotelian method in bioethics

The main claim of this paper is that the method outlined and used in Aristotle's Ethics is an appropriate and credible one to use in bioethics. Here “appropriate” means that the method is capable of establishing claims and developing concepts in bioethics and “credible” that the method has some plausibility, it is not open to obvious and immediate objection. It begins by suggesting why this claim matters and then gives a brief outline of Aristotle's method. The main argument is made in three stages. First, it is argued that Aristotelian method is credible because it compares favourably with alternatives. In this section it is shown that Aristotelian method is not vulnerable to criticisms that are made both of methods that give a primary place to moral theory (such as utilitarianism) and those that eschew moral theory (such as casuistry and social science approaches). As such, it compares favourably with these other approaches that are vulnerable to at least some of these criticisms. Second, the appropriateness of Aristotelian method is indicated through outlining how it would deal with a particular case. Finally, it is argued that the success of Aristotle's philosophy is suggestive of both the credibility and appropriateness of his method.</p

Cirripedia of Madeira

We give a list of Cirripedia from Madeira Island and nearby deep water, based on specimens in the collection of the Museu Municipal do Funchal (Historia Natural) (MMF), records mentioned in the literature, and recent collections. Tesseropora atlantica Newman and Ross, 1976 is recorded from Madeira for the first time. The Megabalanus of Madeira is M. azoricus. There are 20 genera containing 27 species, of which 22 occur in depths less than 200 m. Of these shallow water species, eight are wide-ranging oceanic forms that attach to other organisms or to floating objects, leaving just 13 truly benthic shallow water barnacles. This low diversity is probably a consequence of the distance from the continental coasts and the small area of the available habitat. No endemic species have been found