Sediment Transport Model Including Short-Lived Radioisotopes: Model Description and Idealized Test Cases

Geochronologies derived from sediment cores in coastal locations are often used to infer event bed characteristics such as deposit thicknesses and accumulation rates. Such studies commonly use naturally occurring, short-lived radioisotopes, such as Beryllium-7 (Be-7) and Thorium-234 (Th-234), to study depositional and post-depositional processes. These radioisotope activities, however, are not generally represented in sediment transport models that characterize coastal flood and storm deposition with grain size patterns and deposit thicknesses. We modified the Community Sediment Transport Modeling System (CSTMS) to account for reactive tracers and used this capability to represent the behavior of these short-lived radioisotopes on the sediment bed. This paper describes the model and presents results from a set of idealized, one-dimensional (vertical) test cases. The model configuration represented fluvial deposition followed by periods of episodic storm resuspension. Sensitivity tests explored the influence on seabed radioisotope profiles by the intensities of bioturbation and wave resuspension and the thickness of fluvial deposits. The intensity of biodiffusion affected the persistence of fluvial event beds as evidenced by Be-7. Both resuspension and biodiffusion increased the modeled seabed inventory of Th-234. A thick fluvial deposit increased the seabed inventory of Be-7 and Th-234 but mixing over time greatly reduced the difference in inventory of Th-234 in fluvial deposits of different thicknesses

Analysis of the mtDNA insertion site on chromosome 9L in maize inbreds using fluorescence in situ hybridization

Abstract only availableAlmost all eukaryotic nuclear genomes show evidence of organellar DNA insertions originating from mitochondrial DNA (mtDNA) and chloroplast DNA (cpDNA). While the precise mechanisms of incorporation remain unknown, the phenomenon is frequent and ongoing in many species. In Zea mays, mtDNA insertions differ among inbred lines. A very large mtDNA insertion is found near the centromere of the long arm of chromosome 9 in the B73 inbred. This insertion contains the majority of the mitochondrial genome, while a similarly positioned insertion in the Mo17 inbred line is much smaller. We used recombinant inbred lines from the intermated B73 x Mo17 (IBM) population to determine if the insertions are indeed at the same position. We selected lines with recombination in this region of chromosome 9L. Using two mtDNA probes present in the insertions in both B73 and Mo17, we applied a chromosome painting technique called fluorescence in situ hybridization (FISH) to root-tip metaphase chromosomes and looked for the presence of the mtDNA site on chromosome 9L in the selected IBM lines. If the mtDNA insertion sites in B73 and Mo17 are at different locations, then at least one of the recombinant IBM lines should not display a mtDNA insertion at the chromosome 9 location. However, all of the recombinant IBM lines examined displayed the mtDNA insertion site on chromosome 9L. This indicates that the Mo17 and B73 insertions likely occupy the same region on the chromosome. Furthermore, this suggests that the large mtDNA insertion occurred recently in B73 at a pre-existing site present in both B73 and Mo17.NSF-REU Program in Biological Sciences & Biochemistr

Mitochondrial DNA insertion into nuclear chromosomes of maize

Abstract only availableEvery mitochondrion contains its own DNA separate from the nucleus. Over evolutionary time, most of the mitochondrial genes have moved to the nucleus so that now mitochondria require nuclear DNA to function. This type of transfer is an apparently ongoing process based on our observations that large pieces of the mitochondrial genome have been transferred to the nucleus. The focus of this study was to find the locations of mitochondrial DNA (mtDNA) on nuclear chromosomes in the B73 line of maize and compare these locations to other lines, using fluorescence in situ hybridization (FISH). First, cosmids previously made from a normal mtDNA genotype (NB) were maxi-prepped and then direct labeled with fluorescent tags. Next we prepared slides of B73 root tips and hybridized the labeled cosmids as well as marker probes to the cells. After hybridization, the slides were viewed and chromosome spread pictures were taken showing the location of the cosmids on the chromosomes. This process was then performed on root tip chromosomes from the Mo17, Black Mexican Sweet (BMS), and B37 lines. Twelve cosmids, representing about 71% of the mitochondrial genome, were examined and 8 different nuclear insertion sites were identified. These dispersed locations were predominantly near centromeres or telomeres on chromosomes 2 and 9. These new findings will make understanding the B73 nuclear genome sequence easier because now researchers will know what to expect at these locations and provide new information about the mechanism of mitochondrial genome transfer to the nucleus.NSF-REU Biology & Biochemistr

Identification of chloroplast DNA insertions in nuclear chromosomes of maize B73 line using the FISH procedure

Abstract only availableIt is known that chloroplast DNA can incorporate itself into the nuclear genome of plants. However, the sites of chloroplast (ct) DNA integration into chromosomes of maize have not yet been analyzed. This project is the first attempt to find the location of the ctDNA on the maize chromosomes. Fluorescent in situ hybridization is a technique that has proved useful in karyotyping and chromosomal mapping in maize. The FISH procedure is being used in this study to discover the location of the ctDNA in the nuclear genome of the inbred line B37. In order to develop ctDNA “probes” for FISH analysis, we have used the polymerase chain reaction (PCR) to produce fragments of ctDNA. Primers were chosen to amplify fragments of 10 kb or larger. The amplified DNAs were purified and labeled with fluorescent dyes and these probes were subsequently hybridized to chromosomes. The probes recognize and bind to the corresponding DNA sequences within the chromosomes. Root tip cells were used to prepare the slides for hybridization. Because the cells are collected during the metaphase stage of division, the chromosomes are compact and more easily visible. Chromosomes that contain ctDNA can be detected using a compound microscope with fluorescent attachments. The location of the ctDNA on the chromosomes is made visible by the fluorescent labeling of the probe. Eight of eleven regions of the chloroplast genome of the B73 line have been specifically amplified and have been observed under the microscope for FISH analysis. This information will contribute to an understanding of the extent and mechanism of transfer of organellar genomes to the nucleus.MU Monsanto Undergraduate Research Fellowshi

Analysis of mitochondrial DNA insertions into a nuclear chromosome of the maize B73 line

Abstract only availableMitochondrial DNA (mtDNA) is known to have integrated into the nuclear DNA of plants and animals. The purpose of this project is to investigate the on-going migration of mtDNA into the nuclear DNA of maize plants. Specific objectives are to discover the amount of DNA incorporated, whether it is the whole mitochondrial genome or sections, and to see if it has replicated after migration. The maize inbred line B73 has a particularly large mt DNA insert on chromosome 9. Using the fluorescent in situ hybridization (FISH) method, the arrangement of inserted mitochondrial DNA was examined. The FISH method uses fluorescently labeled mtDNA as probes for hybridization to chromosomes. Regions of the chromosomes that contain mtDNA can then be detected using a compound microscope with fluorescent attachments. Locations that contain more mtDNA are brighter. Three combinations of probes that cover different parts of the mitochondrial genome were employed. In order to analyze the arrangement of the DNA, the chromosomes were prepared from a stage of meiosis called pachynema in which the chromosomes are elongated and have not yet begun to condense. The results have confirmed the presence of all three probes within the large insertion of mtDNA on chromosome 9 of B73. The data suggest that either different parts of the mitochondrial genome are incorporated preferentially or that there is selective replication of portions of the mitochondrial genome after incorporation.MU Monsanto Undergraduate Research Fellowshi

Parallel altitudinal clines reveal trends in adaptive evolution of genome size in \u3ci\u3eZea mays\u3c/i\u3e

While the vast majority of genome size variation in plants is due to differences in repetitive sequence, we know little about how selection acts on repeat content in natural populations. Here we investigate parallel changes in intraspecific genome size and repeat content of domesticated maize (Zea mays) landraces and their wild relative teosinte across altitudinal gradients in Mesoamerica and South America. We combine genotyping, low coverage whole-genome sequence data, and flow cytometry to test for evidence of selection on genome size and individual repeat abundance. We find that population structure alone cannot explain the observed variation, implying that clinal patterns of genome size are maintained by natural selection. Our modeling additionally provides evidence of selection on individual heterochromatic knob repeats, likely due to their large individual contribution to genome size. To better understand the phenotypes driving selection on genome size, we conducted a growth chamber experiment using a population of highland teosinte exhibiting extensive variation in genome size. We find weak support for a positive correlation between genome size and cell size, but stronger support for a negative correlation between genome size and the rate of cell production. Reanalyzing published data of cell counts in maize shoot apical meristems, we then identify a negative correlation between cell production rate and flowering time. Together, our data suggest a model in which variation in genome size is driven by natural selection on flowering time across altitudinal clines, connecting intraspecific variation in repetitive sequence to important differences in adaptive phenotypes

Aluminum tolerance in maize is associated with higher MATE1 gene copy number

Genome structure variation, including copy number variation and presence/absence variation, comprises a large extent of maize genetic diversity; however, its effect on phenotypes remains largely unexplored. Here, we describe how copy number variation underlies a rare allele that contributes to maize aluminum (Al) tolerance. Al toxicity is the primary limitation for crop production on acid soils, which make up 50% of the world's potentially arable lands. In a recombinant inbred line mapping population, copy number variation of the Al tolerance gene multidrug and toxic compound extrusion 1 (MATE1) is the basis for the quantitative trait locus of largest effect on phenotypic variation. This expansion in MATE1 copy number is associated with higher MATE1 expression, which in turn results in superior Al tolerance. The three MATE1 copies are identical and are part of a tandem triplication. Only three maize inbred lines carrying the three-copy allele were identified from maize and teosinte diversity panels, indicating that copy number variation for MATE1 is a rare, and quite likely recent, event. These maize lines with higher MATE1 copy number are also Al-tolerant, have high MATE1 expression, and originate from regions of highly acidic soils. Our findings show a role for copy number variation in the adaptation of maize to acidic soils in the tropics and suggest that genome structural changes may be a rapid evolutionary response to new environments