Genetic characterization of the Cy transposable element system at the Bz locus of Zea mays L.

The unstable bz-rcy allele arose by the insertion of a receptor element, rcy, into the Bz locus of a single gamete from the TEL population. Mutability of bz-rcy is controlled by the independently segregating regulatory element Cy (Cycler). In the absence of Cy, bz-rcy conditions a stable bronze aleurone. In the presence of Cy, bz-rcy conditions many small fully colored spots on a bronze background. These two elements, rcy and Cy, were previously undescribed;Genetic tests have established a relationship between Cy and the Mutator system. Cy is not functionally homologous to any of the non-Mutator transposable element systems;The number of genetically active Cy elements in a plant can increase or decrease via Cy transposition;Nonresponsive derivatives of bz-rcy have been isolated. A model has been established to explain the loss of the distal markers C and Sh coincident with the origin of some of these derivatives;The original isolate of bz-rcy often generates derivative alleles (states) that condition altered spotting patterns, e.g., reduced numbers of spots and/or larger and smaller spots, that reflect alterations in the frequency and timing of rcy excisions from bz-rcy. In contrast to receptors of the Ac and En(Spm) systems, which undergo changes of state only in the presence of the appropriate regulatory element, bz-rcy can undergo changes of state in the absence of an active Cy;Cy changes of state occur less frequently than those of bz-rcy. States of Cy have been isolated that induce reduced rates (but not altered timing) of rcy excision from bz-rcy. Many of the Cy states show progressive loss of function over succeeding generations;Many bz-rcy states and some Cy states have the ability to revert to fine-high spotting (cycling). Cycling is developmentally regulated and occurs only in the presence of Cy. A model has been proposed which evokes a form of reversible DNA modification to explain the cycling phenomenon and the origin of bz-rcy states in the absence of Cy;Of 47 diverse maize lines assayed, only two sources were found to contain strong Cy elements, i.e., Mutator-related stocks and the TEL population. Of the remaining lines, six, contained weak Cy elements and the rest lacked genetically detectable Cy elements

RNA-Seq Based Analysis of Population Structure within the Maize Inbred B73

Recent reports have shown than many identically named genetic lines used in research around the world actually contain large amounts of uncharacterized genetic variation as a result of cross contamination of stocks, unintentional crossing, residual heterozygosity within original stocks, or de novo mutation. 27 public, large scale, RNA-seq datasets from 20 independent research groups around the world were used to assess variation within the maize (Zea mays ssp. mays) inbred B73, a four decade old variety which served as the reference genotype for the original maize genome sequencing project and is widely used in genetic, genomic, and phenotypic research. Several clearly distinct clades were identified among putatively B73 samples. A number of these clades were defined by the presence of clearly defined genomic blocks containing a haplotype which did not match the published B73 reference genome. The overall proportion of the maize genotype where multiple distinct haplotypes were observed across different research groups was approximately 2.3%. In some cases the relationship among B73 samples generated by different research groups recapitulated mentor/mentee relationships within the maize genetics community

Genes Identified by Visible Mutant Phenotypes Show Increased Bias toward One of Two Subgenomes of Maize

Not all genes are created equal. Despite being supported by sequence conservation and expression data, knockout homozygotes of many genes show no visible effects, at least under laboratory conditions. We have identified a set of maize (Zea mays L.) genes which have been the subject of a disproportionate share of publications recorded at MaizeGDB. We manually anchored these “classical” maize genes to gene models in the B73 reference genome, and identified syntenic orthologs in other grass genomes. In addition to proofing the most recent version 2 maize gene models, we show that a subset of these genes, those that were identified by morphological phenotype prior to cloning, are retained at syntenic locations throughout the grasses at much higher levels than the average expressed maize gene, and are preferentially found on the maize1 subgenome even with a duplicate copy is still retained on the opposite subgenome. Maize1 is the subgenome that experienced less gene loss following the whole genome duplication in maize lineage 5–12 million years ago and genes located on this subgenome tend to be expressed at higher levels in modern maize. Links to the web based software that supported our syntenic analyses in the grasses should empower further research and support teaching involving the history of maize genetic research. Our findings exemplify the concept of “grasses as a single genetic system,” where what is learned in one grass may be applied to another

High Density Genetic Maps of Seashore Paspalum Using Genotyping-By-Sequencing and Their Relationship to The Sorghum Bicolor Genome.

As a step towards trait mapping in the halophyte seashore paspalum (Paspalum vaginatum Sw.), we developed an F1 mapping population from a cross between two genetically diverse and heterozygous accessions, 509022 and HI33. Progeny were genotyped using a genotyping-by-sequencing (GBS) approach and sequence reads were analyzed for single nucleotide polymorphisms (SNPs) using the UGbS-Flex pipeline. More markers were identified that segregated in the maternal parent (HA maps) compared to the paternal parent (AH maps), suggesting that 509022 had overall higher levels of heterozygosity than HI33. We also generated maps that consisted of markers that were heterozygous in both parents (HH maps). The AH, HA and HH maps each comprised more than 1000 markers. Markers formed 10 linkage groups, corresponding to the ten seashore paspalum chromosomes. Comparative analyses showed that each seashore paspalum chromosome was syntenic to and highly colinear with a single sorghum chromosome. Four inversions were identified, two of which were sorghum-specific while the other two were likely specific to seashore paspalum. These high-density maps are the first available genetic maps for seashore paspalum. The maps will provide a valuable tool for plant breeders and others in the Paspalum community to identify traits of interest, including salt tolerance

Last Frontier of Agricultural Big Data in Rotation?

Over the years new and promising varieties are bred not only for resistance to diseases but also for direct yield by stress tolerance, architectural or other properties. However, these potential yield improvements under optimal circumstances are not met by the growers in their practice, and is even widening. ..

tGBS® genotyping-by-sequencing enables reliable genotyping of heterozygous loci

Conventional genotyping-by-sequencing (cGBS) strategies suffer from high rates of missing data and genotyping errors, particularly at heterozygous sites. tGBS® genotyping-by-sequencing is a novel method of genome reduction that employs two restriction enzymes to generate overhangs in opposite orientations to which (single-strand) oligos rather than (double-stranded) adaptors are ligated. This strategy ensures that only doubledigested fragments are amplified and sequenced. The use of oligos avoids the necessity of preparing adaptors and the problems associated with inter-adaptor annealing/ligation. Hence, the tGBS protocol simplifies the preparation of high-quality GBS sequencing libraries. During polymerase chain reaction (PCR) amplification, selective nucleotides included at the 3\u27-end of the PCR primers result in additional genome reduction as compared to cGBS. By adjusting the number of selective bases, different numbers of genomic sites are targeted for sequencing. Therefore, for equivalent amounts of sequencing, more reads per site are available for SNP calling. Hence, as compared to cGBS, tGBS delivers higher SNP calling accuracy (\u3e97–99%), even at heterozygous sites, less missing data per marker across a population of samples, and an enhanced ability to genotype rare alleles. tGBS is particularly well suited for genomic selection, which often requires the ability to genotype populations of individuals that are heterozygous at many loci

Optimising the identification of causal variants across varying genetic architectures in crops

Association studies use statistical links between genetic markers and the phenotype variation across many individuals to identify genes controlling variation in the target phenotype. However, this approach, particularly conducted on a genome-wide scale (GWAS), has limited power to identify the genes responsible for variation in traits controlled by complex genetic architectures. In this study, we employ real-world genotype datasets from four crop species with distinct minor allele frequency distributions, population structures and linkage disequilibrium patterns. We demonstrate that different GWAS statistical approaches provide favourable trade-offs between power and accuracy for traits controlled by different types of genetic architectures. FarmCPU provides the most favourable outcomes for moderately complex traits while a Bayesian approach adopted from genomic prediction provides the most favourable outcomes for extremely complex traits. We assert that by estimating the complexity of genetic architectures for target traits and selecting an appropriate statistical approach for the degree of complexity detected, researchers can substantially improve the ability to dissect the genetic factors controlling complex traits such as flowering time, plant height and yield component

Functional Modeling of Plant Growth Dynamics

Recent advances in automated plant phenotyping have enabled the collection of time series measurements from the same plants of a wide range of traits at different developmental time scales. The availability of time series phenotypic datasets has increased interest in statistical approaches for comparing patterns of change among different plant genotypes and different treatment conditions. Two widely used methods of modeling growth with time are pointwise analysis of variance (ANOVA) and parametric sigmoidal curve fitting. Pointwise ANOVA yields discontinuous growth curves, which do not reflect the true dynamics of growth patterns in plants. In contrast, fitting a parametric model to a time series of observations does capture the trend of growth; however, these models require assumptions regarding the true pattern of plant growth. Depending on the species, treatment regime, and subset of the plant life cycle sampled, these assumptions will not always hold true. We have developed a different approach—functional ANOVA—which yields continuous growth curves without requiring assumptions regarding patterns of plant growth. We compared and validated this approach using data from an experiment measuring the growth of two maize (Zea mays L. ssp. mays) genotypes under two water availability treatments during a 21-d period. Functional ANOVA enables a nonparametric estimation of the dynamics of changes in plant traits with time without assumptions regarding curve shape. In addition to estimating smooth curves of trait values with time, functional ANOVA also estimates the derivatives of these curves, e.g., growth rates, simultaneously. Using two different subsampling strategies, we demonstrate that this functional ANOVA method enables the comparison of growth curves among plants phenotyped on non-overlapping days with little reduction in estimation accuracy. This means that functional ANOVA based approaches can allow larger numbers of samples and biological replicates to be scored in a single experiment given fixed amounts of phenotyping infrastructure and personnel

Genetic control of photoprotection and photosystem II operating efficiency in plants

• Photoprotection against excess light via nonphotochemical quenching (NPQ) is indispensable for plant survival. However, slow NPQ relaxation under low light conditions can decrease yield of field-grown crops up to 40%. • Using semi-high-throughput assay, we quantified the kinetics of NPQ and photosystem II operating efficiency (ΦPSII) in a replicated field trial of more than 700 maize (Zea mays) genotypes across 2 yr. Parametrized kinetics data were used to conduct genome-wide association studies. • For six candidate genes involved in NPQ and ΦPSII kinetics in maize the loss of function alleles of orthologous genes in Arabidopsis (Arabidopsis thaliana) were characterized: two thioredoxin genes, and genes encoding a transporter in the chloroplast envelope, an initiator of chloroplast movement, a putative regulator of cell elongation and stomatal patterning, and a protein involved in plant energy homeostasis. • Since maize and Arabidopsis are distantly related, we propose that genes involved in photoprotection and PSII function are conserved across vascular plants. The genes and naturally occurring functional alleles identified here considerably expand the toolbox to achieving a sustainable increase in crop productivity