Maria G. Salas Fernandez

Rare genetic variation at Zea mays crtRB1 increases Β-carotene in maize grain

Breeding to increase β-carotene levels in cereal grains, termed provitamin A biofortification, is an economical approach to address dietary vitamin A deficiency in the developing world. Experimental evidence from association and linkage populations in maize (Zea mays L.) demonstrate that the gene encoding β-carotene hydroxylase 1 (crtRB1) underlies a principal quantitative trait locus associated with β-carotene concentration and conversion in maize kernels. crtRB1 alleles associated with reduced transcript expression correlate with higher β-carotene concentrations. Genetic variation at crtRB1 also affects hydroxylation efficiency among encoded allozymes, as observed by resultant carotenoid profiles in recombinant expression assays. The most favorable crtRB1 alleles, rare in frequency and unique to temperate germplasm, are being introgressed via inexpensive PCR marker-assisted selection into tropical maize germplasm adapted to developing countries, where it is most needed for human health.

From dwarves to giants? Plant height manipulation for biomass yield.

The increasing demand for lignocellulosic biomass for the production of biofuels provides value to vegetative plant tissue and leads to a paradigm shift for optimizing plant architecture in bioenergy crops. Plant height (PHT) is among the most important biomass yield components and is the focus of this review, with emphasis on the energy grasses maize (Zea mays) and sorghum (Sorghum bicolor). We discuss the scientific advances in the identification of PHT quantitative trait loci (QTLs) and the understanding of pathways and genes controlling PHT, especially gibberellins and brassinosteroids. We consider pleiotropic effects of QTLs or genes affecting PHT on other agronomically important traits and, finally, we discuss strategies for applying this knowledge to the improvement of dual-purpose or dedicated bioenergy crops.

Equilibrium Processes Cannot Explain High Levels of Short- and Medium-Range Linkage Disequilibrium in the Domesticated Grass Sorghum bicolor

Patterns of linkage disequilibrium (LD) are of interest because they provide evidence of both equilibrium (e.g., mating system or long-term population structure) and nonequilibrium (e.g., demographic or selective) processes, as well as because of their importance in strategies for identifying the genetic basis of complex phenotypes. We report patterns of short and medium range (up to100 kb) LD in six unlinked genomic regions in the partially selfing domesticated grass, Sorghum bicolor. The extent of allelic associations in S. bicolor, as assessed by pairwise measures of LD, is higher than in maize but lower than in Arabidopsis, in qualitative agreement with expectations based on mating system. Quantitative analyses of the population recombination parameter, ρ, however, based on empirical estimates of rates of recombination, mutation, and self-pollination, show that LD is more extensive than expected under a neutral equilibrium model. The disparity between ρ and the population mutation parameter, θ, is similar to that observed in other species whose population history appears to be complex. From a practical standpoint, these results suggest that S. bicolor is well suited for association studies using reasonable numbers of markers, since LD typically extends at least several kilobases but has largely decayed by 15 kb.

Genome-wide association study for nine plant architecture traits in Sorghum

The 101 SNPs were associated with at least one of nine plant architecture traits KS3 gene was associated with variation in seed number GA2ox5 gene included in a significant region on chromosome 9 controlling plant height Novel genomic regions were associated with stem circumference and internode number Novel genomic regions were associated with tiller number, panicle exsertion, and length

A high-throughput, field-based phenotyping technology for tall biomass crops

A ground-based platform enables high-throughput, image-based data and analysis to characterize plant architecture of a tall biomass crop species. Recent advances in omics technologies have not been accompanied by equally efficient, cost-effective, and accurate phenotyping methods required to dissect the genetic architecture of complex traits. Even though high-throughput phenotyping platforms have been developed for controlled environments, field-based aerial and ground technologies have only been designed and deployed for short-stature crops. Therefore, we developed and tested Phenobot 1.0, an auto-steered and self-propelled field-based high-throughput phenotyping platform for tall dense canopy crops, such as sorghum (Sorghum bicolor). Phenobot 1.0 was equipped with laterally positioned and vertically stacked stereo RGB cameras. Images collected from 307 diverse sorghum lines were reconstructed in 3D for feature extraction. User interfaces were developed, and multiple algorithms were evaluated for their accuracy in estimating plant height and stem diameter. Tested feature extraction methods included the following: (1) User-interactive Individual Plant Height Extraction (UsIn-PHe) based on dense stereo three-dimensional reconstruction; (2) Automatic Hedge-based Plant Height Extraction (Auto-PHe) based on dense stereo 3D reconstruction; (3) User-interactive Dense Stereo Matching Stem Diameter Extraction; and (4) User-interactive Image Patch Stereo Matching Stem Diameter Extraction (IPaS-Di). Comparative genome-wide association analysis and ground-truth validation demonstrated that both UsIn-PHe and Auto-PHe were accurate methods to estimate plant height, while Auto-PHe had the additional advantage of being a completely automated process. For stem diameter, IPaS-Di generated the most accurate estimates of this biomass-related architectural trait. In summary, our technology was proven robust to obtain ground-based high-throughput plant architecture parameters of sorghum, a tall and densely planted crop species.

Genetic architecture of photosynthesis in Sorghum bicolor under non-stress and cold stress conditions

Multiple genomic regions explain variation in the carbon fixation response to non-stress and cold stress in Sorghum bicolor, with some of these regions simultaneously controlling multiple traits.

Differential manipulation of leaf angle throughout the canopy: current status and prospects

Leaf angle is defined as the inclination between the midrib of the leaf blade and the vertical stem of a plant. This trait has been identified as a key component in the development of high-yielding varieties of cereal species, particularly maize, rice, wheat, and sorghum. The effect of leaf angle on light interception efficiency, photosynthetic rate, and yield has been investigated since the 1960s, yet, significant knowledge gaps remain in understanding the genetic control of this complex trait. Recent advances in physiology and modeling have proposed a plant ideotype with varying leaf angles throughout the canopy. In this context, we present historical and recent evidence of: (i) the effect of leaf angle on photosynthetic efficiency and yield; (ii) the hormonal regulation of this trait; (iii) the current knowledge on its quantitative genetic control; and (iv) the opportunity to utilize high-throughput phenotyping methods to characterize leaf angle at multiple canopy levels. We focus on research conducted on grass species of economic importance, with similar plant architecture and growth patterns. Finally, we present the challenges and strategies plant breeders will need to embrace in order to manipulate leaf angle differentially throughout the canopy and develop superior crops for food, feed, and fuel production.

A cost-effective and customizable automated irrigation system for precise high-throughput phenotyping in drought stress studies.

The development of high-yielding crops with drought tolerance is necessary to increase food, feed, fiber and fuel production. Methods that create similar environmental conditions for a large number of genotypes are essential to investigate plant responses to drought in gene discovery studies. Modern facilities that control water availability for each plant remain cost-prohibited to some sections of the research community. We present an alternative cost-effective automated irrigation system scalable for a high-throughput and controlled dry-down treatment of plants. This system was tested in sorghum using two experiments. First, four genotypes were subjected to ten days of dry-down to achieve three final Volumetric Water Content (VWC) levels: drought (0.10 and 0.20 m3 m-3) and control (0.30 m3 m-3). The final average VWC was 0.11, 0.22, and 0.31 m3 m-3, respectively, and significant differences in biomass accumulation were observed between control and drought treatments. Second, 42 diverse sorghum genotypes were subjected to a seven-day dry-down treatment for a final drought stress of 0.15 m3 m-3 VWC. The final average VWC was 0.17 m3 m-3, and plants presented significant differences in photosynthetic rate during the drought period. These results demonstrate that cost-effective automation systems can successfully control substrate water content for each plant, to accurately compare their phenotypic responses to drought, and be scaled up for high-throughput phenotyping studies.

Field-based robotic phenotyping of sorghum plant architecture using stereo vision: BAO et al.

Funding information National Institute of Food and Agriculture, Grant/Award Number: 2012‐67009‐19713; United States Department of Agriculture Abstract Sorghum (Sorghum bicolor) is known as a major feedstock for biofuel production. To improve its biomass yield through genetic research, manually measuring yield component traits (e.g. plant height, stem diameter, leaf angle, leaf area, leaf number, and panicle size) in the field is the current best practice. However, such laborious and time‐consuming tasks have become a bottleneck limiting experiment scale and data acquisition frequency. This paper presents a high‐throughput field‐based robotic phenotyping system which performed side‐view stereo imaging for dense sorghum plants with a wide range of plant heights throughout the growing season. Our study demonstrated the suitability of stereo vision for field‐based three‐dimensional plant phenotyping when recent advances in stereo matching algorithms were incorporated. A robust data processing pipeline was developed to quantify the variations or morphological traits in plant architecture, which included plot‐based plant height, plot‐based plant width, convex hull volume, plant surface area, and stem diameter (semiautomated). These image‐derived measurements were highly repeatable and showed high correlations with the in‐field manual measurements. Meanwhile, manually collecting the same traits required a large amount of manpower and time compared to the robotic system. The results demonstrated that the proposed system could be a promising tool for large‐scale field‐based high‐throughput plant phenotyping of bioenergy crops.

Assessment of genetic diversity among sorghum landraces and their wild/weedy relatives in western Kenya using simple sequence repeat (SSR) markers

Understanding the extent of gene exchange between cultivated sorghum and its wild/weedy relatives and the evolutionary processes (including farmers’ practices) that act to shape the structure of genetic diversity within and between them is an important aspect for germplasm conservation strategies, biosafety risk assessment, and crop improvement programs. In this study, molecular characterization and genetic diversity analyses were conducted on wild, weedy and cultivated sorghums collected at a local-scale in a traditional farming system in the Lambwe Valley of western Kenya. Nine simple sequence repeat (SSR) markers were used to genotype 294 cultivated sorghum and 200 wild sorghum individuals. The nine SSR markers were highly polymorphic with a number of alleles that varied from 2 to 19. Overall, wild sorghums had higher genetic diversity, observed heterozygosity, total number of alleles, polymorphic information content and more genotypes per locus than the cultivated types. A Mantel test demonstrated that there was significant isolation-by-distance for wilds and cultivated materials. STRUCTURE, cluster and principal coordinate analyses consistently assigned wild and cultivated individuals to different groups but failed to place hybrids/weedy types as a single separate group from wilds. Our results provide strong evidence of significant genetic diversity retained within wilds, larger divergence between wild and cultivated materials and reduced gene flow than those previously reported in Kenya. These results demonstrate the value of the Lambwe Valley region as a genetic reservoir and the importance to conduct genetic diversity studies at the local scale to design and execute appropriate in situ conservation programs and policies.