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Allen D.K.,Plant Genetics Research Unit | Allen D.K.,Donald Danforth Plant Science Center
Current Opinion in Biotechnology | Year: 2016

Analyses of metabolic flux using stable isotopes in plants have traditionally been restricted to tissues with presumed homogeneous cell populations and long metabolic steady states such as developing seeds, cell suspensions, or cultured roots and root tips. It is now possible to describe these and other metabolically more dynamic tissues such as leaves in greater detail using novel methods in mass spectrometry, isotope labeling strategies, and transient labeling-based flux analyses. Such studies are necessary for a systems level description of plant function that more closely represents biological reality, and provides insights into the genes that will need to be modified as natural resources become ever more limited and environments change. © 2015.

Shyama Prasad Rao R.,University of Missouri | Thelen J.J.,University of Missouri | Miernyk J.A.,University of Missouri | Miernyk J.A.,Plant Genetics Research Unit
Frontiers in Plant Science | Year: 2014

Among post-translational modifications, there are some conceptual similarities between Lys-Nε-acetylation and Ser/Thr/Tyr O-phosphorylation. Herein we present a bioinformatics-based overview of reversible protein Lys-acetylation, including some comparisons with reversible protein phosphorylation. The study of Lys-acetylation of plant proteins has lagged behind studies of mammalian and microbial cells; 1000s of acetylation sites have been identified in mammalian proteins compared with only hundreds of sites in plant proteins. While most previous emphasis was focused on post-translational modifications of histones, more recent studies have addressed metabolic regulation. Being directly coupled with cellular CoA/acetyl-CoA and NAD/NADH, reversible Lys-Nε-acetylation has the potential to control, or contribute to control, of primary metabolism, signaling, and growth and development. © 2014 Rao, Thelen and Miernyk.

Becker A.,Donald Danforth Plant Science Center | Chao D.-Y.,Purdue University | Zhang X.,University of Chicago | Salt D.E.,Purdue University | And 2 more authors.
PLoS ONE | Year: 2011

Bulk segregant analysis (BSA) using microarrays, and extreme array mapping (XAM) have recently been used to rapidly identify genomic regions associated with phenotypes in multiple species. These experiments, however, require the identification of single feature polymorphisms (SFP) between the cross parents for each new combination of genotypes, which raises the cost of experiments. The availability of the genomic polymorphism data in Arabidopsis thaliana, coupled with the efficient designs of Single Nucleotide Polymorphism (SNP) genotyping arrays removes the requirement for SFP detection and lowers the per array cost, thereby lowering the overall cost per experiment. To demonstrate that these approaches would be functional on SNP arrays and determine confidence intervals, we analyzed hybridizations of natural accessions to the Arabidopsis ATSNPTILE array and simulated BSA or XAM given a variety of gene models, populations, and bulk selection parameters. Our results show a striking degree of correlation between the genotyping output of both methods, which suggests that the benefit of SFP genotyping in context of BSA can be had with the cheaper, more efficient SNP arrays. As a final proof of concept, we hybridized the DNA from bulks of an F2 mapping population of a Sulfur and Selenium ionomics mutant to both the Arabidopsis ATTILE1R and ATSNPTILE arrays, which produced almost identical results. We have produced R scripts that prompt the user for the required parameters and perform the BSA analysis using the ATSNPTILE1 array and have provided them as supplemental data files.

Vincent J.A.,University of Missouri | Stacey M.,University of Missouri | Stacey G.,University of Missouri | Bilyeu K.D.,University of Missouri | Bilyeu K.D.,Plant Genetics Research Unit
Plant Genome | Year: 2015

Soybean seeds contain a large amount of P, which is stored as phytic acid (PA). Phytic acid is indigestible by nonruminant livestock and considered an antinutritional factor in soybean meal. Several low PA soybean lines have been discovered, but many of these lines have either minor reductions in PA or inadequate germination and emergence. The reduced PA phenotype of soybean line Gm-lpa-ZC-2 was previously shown to be the result of a mutation in a gene encoding an inositol pentakisphosphate 2-kinase on chromosome 14 (14IPK1). While the 14IPK1 mutation was shown to have no impact on germination and emergence, the reduction in PA was modest (up to 50%). Our objective was to determine the effect on seed P partitioning for a novel mutation of an independent IPK1 gene on chromosome six (06IPK1) on its own and in combination with mutant alleles of the 14IPK1. We developed soybean populations and conducted genotype and phenotype association analyses based on the genotype of the 06IPK1 and 14IPK1 genes and the seed P partitioning profile. The lines with both mutant IPK1 genes had very low PA levels, moderate accumulation of inorganic phosphate (Pi), and accumulation of high amounts of P in lower inositols. The developed lines did not have significant reductions in germination or field emergence. In addition, characterization of the lower inositols produced in the mutant lines suggests that IPK1 is a polyphosphate kinase and provides some insight into the PA biosynthesis pathway in soybean seeds. © Crop Science Society of America.

Krishnan H.B.,Plant Genetics Research Unit | Krishnan H.B.,University of Missouri | Wang T.T.Y.,U.S. Department of Agriculture
Food Chemistry | Year: 2015

Lunasin is a 5-kDa soybean bioactive peptide with demonstrated anti-cancer and anti-inflammatory properties. Recently, purification methods have been developed to obtain gram quantities of lunasin. However, these methods are cumbersome, time consuming and cost-prohibitive. To overcome these constrains we have developed a novel method which involves extraction of soybean flour with 30% ethanol followed by preferential precipitation of lunasin and protease inhibitors by calcium. The calcium precipitated protein fraction, which we termed as Lunasin Protease Inhibitor Concentrate (LPIC), contains three abundant proteins with molecular weights of 21, 14 and 5 kDa. This simple procedure yields 3.2 g of LPIC from 100 g of soybean flour and the entire isolation procedure can be completed in less than 2 h. Treatment of THP-1 human monocyte cell lines with LPIC resulted in suppression of lipopolysaccharide-stimulated cytokine expression, demonstrating that the LPIC isolated by our simple procedure is biologically active.

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