Gene Expression Profiling of Iron Deficiency Chlorosis Sensitive and Tolerant Soybean Indicates Key Roles for Phenylpropanoids under Alkalinity Stress

Alkaline soils comprise 30% of the earth and have low plant-available iron (Fe) concentration, and can cause iron deficiency chlorosis (IDC). IDC causes soybean yield losses of $260 million annually. However, it is not known whether molecular responses to IDC are equivalent to responses to low iron supply. IDC tolerant and sensitive soybean lines provide a contrast to identify specific factors associated with IDC.We used RNA-seq to compare gene expression under combinations of normal pH (5.7) or alkaline pH (7.7, imposed by 2.5mM bicarbonate, or pH 8.2 imposed by 5mM bicarbonate) and normal (25μM) or low (1μM) iron conditions from roots of these lines. Thus, we were able to treat pH and Fe supply as separate variables. We also noted differential gene expression between IDC sensitive and tolerant genotypes in each condition. Classical iron uptake genes, including ferric-chelate reductase (FCR) and ferrous transporters, were upregulated by both Fe deficiency and alkaline stress, however, their gene products did not function well at alkaline pH. In addition, genes in the phenylpropanoid synthesis pathway were upregulated in both alkaline and low Fe conditions. These genes lead to the production of fluorescent root exudate (FluRE) compounds, such as coumarins. Fluorescence of nutrient solution increased with alkaline treatment, and was higher in the IDC tolerant line. Some of these genes also localized to previously identified QTL regions associated with IDC. We hypothesize that FluRE become essential at alkaline pH where the classical iron uptake system does not function well. This work could result in new strategies to screen for IDC tolerance, and provide breeding targets to improve crop alkaline stress tolerance

Response of \u3ci\u3eAmaranthus\u3c/i\u3e spp. following exposure to sublethal herbicide rates via spray particle drift

The adverse consequences of herbicide drift towards sensitive crops have been extensively reported in the literature. However, little to no information is available on the consequences of herbicide drift onto weed species inhabiting boundaries of agricultural fields. Exposure to herbicide drift could be detrimental to long-term weed management as several weed species have evolved herbicide-resistance after recurrent selection with sublethal herbicide rates This study investigated the deposition of glyphosate, 2,4-D, and dicamba spray particle drift from applications with two different nozzles in a low speed wind tunnel, and their impact on growth and development of Amaranthus spp. Herbicide drift resulted in biomass reduction or complete plant mortality. Inflection points (distance to 50% biomass reduction) for Amaranthus tuberculatus were 7.7, 4.0, and 4.1 m downwind distance for glyphosate, 2,4-D, and dicamba applications with the flat-fan nozzle, respectively, whereas these values corresponded to 2.8, 2.5, and 1.9 m for applications with the air-inclusion nozzle. Inflection points for Amaranthus palmeri biomass reduction were 16.3, 10.9, and 11.5 m for glyphosate, 2,4-D, and dicamba applications with the flat-fan nozzle, respectively, whereas these values corresponded to 7.6, 5.4, and 5.4 m for applications with the air-inclusion nozzle. Plants were more sensitive to glyphosate at higher exposure rates than other herbicides, whereas plants were more sensitive to 2,4-D and dicamba at lower exposure rates compared to glyphosate. Applications with the flat-fan nozzle resulted in 32.3 and 11.5% drift of the applied rate at 1.0 and 3.0 m downwind, respectively, whereas the air-inclusion nozzle decreased the dose exposure in the same distances (11.4 and 2.7%, respectively). Herbicide drift towards field boundaries was influenced by nozzle design and exposed weeds to herbicide rates previously reported to select for herbicide-resistant biotypes

Evaluation of Greenbug and Yellow Sugarcane Aphid Feeding Behavior on Resistant and Susceptible Switchgrass Cultivars

Switchgrass (Panicum virgatum L.) is an emerging biofuel crop that serves as host for aphids. To discern the effects of plant age and possible resistance mechanisms, the feeding behavior of greenbugs (Schizaphis graminum Rondani.) and the yellow sugarcane aphid (Sipha flava Forbes.) was monitored on three diverse switchgrasses by the electrical penetration graph (EPG) technique. Callose deposition and genes associated with callose metabolism were also analyzed to discern their association with plant resistance. There was a strong host effect on greenbugs feeding on lowland cultivar Kanlow at the V3 stage of development, as compared to the greenbug-susceptible upland cultivar Summer and plants derived from Kanlow (♂) × Summer (♀) (K×S) crosses. These data confirmed that Kanlow at the V3 stage had antibiosis to greenbugs, which was absent in the Summer and K×S plants. In contrast, similar effects were not observed for yellow sugarcane aphids, excluding significant differences in the time to first probe on Kanlow plants at the V1 stage and reduction in time spent on pathway processes on Kanlow plants at the V3 stage. These data demonstrated that Kanlow plants may have multiple sources of resistance to the two aphids, and possibly some were phloem based. Microscopy of leaf sections stained with aniline blue for callose was suggestive of increased callose deposition in the sieve elements in Kanlow plants relative to Summer and K×S plants. RT-qPCR analysis of several genes associated with callose metabolism in infested plants was equivocal. Overall, these studies suggest the presence of multiple defense mechanisms against aphids in Kanlow plants, relative to Summer and K×S plants

Two fingerprinting sets for Humulus lupulus based on KASP and microsatellite markers

Verification of clonal identity of hop (Humulus lupulus L.) cultivars within breeding programs and germplasm collections is vital to conserving genetic resources. Accurate and economic DNA-based tools are needed in dioecious hop to confirm identity and parentage, neither of which can be reliably determined from morphological observations. In this study, we developed two fingerprinting sets for hop: a 9-SSR fingerprinting set containing high-core repeats that can be run in a single PCR reaction and a kompetitive allele specific PCR (KASP) assay of 25 single nucleotide polymorphisms (SNPs). The SSR set contains a sex-linked primer pair, HI-AGA7, that was used to genotype 629 hop accessions from the US Department of Agriculture (USDA) National Clonal Germplasm Repository (NCGR), the USDA Forage Seed and Cereal Research (FSCR), and the University of Nebraska-Lincoln (UNL) collections. The SSR set identified unique genotypes except for 89 sets of synonymous samples. These synonyms included: cultivars with different designations, the same cultivars from different sources, heat-treated clones, and clonal variants. Population structure analysis clustered accessions into wild North American (WNA) and cultivated groups. Diversity was slightly higher in the cultivated samples due to larger sample size. Parentage and sib-ship analyses were used to identify true-to-type cultivars. The HI-AGA7 marker generated two male- and nine female-specific alleles among the cultivated and WNA samples. The SSR and KASP fingerprinting sets were compared in 190 samples consisting of cultivated and WNA accession for their ability to confirm identity and assess diversity and population structure. The SSR fingerprinting set distinguished cultivars, selections and WNA accessions while the KASP assays were unable to distinguish the WNA samples and had lower diversity estimates than the SSR set. Both fingerprinting sets are valuable tools for identity confirmation and parentage analysis in hop for different purposes. The 9-SSR assay is cost efficient when genotyping a small number of wild and cultivated hop samples (\u3c96) while the KASP assay is easy to interpret and cost efficient for genotyping a large number of cultivated samples (multiples of 96)

Albumin and mammalian cell culture: implications for biotechnology applications

Albumin has a long historical involvement in design of media for the successful culture of mammalian cells, in both the research and commercial fields. The potential application of albumins, bovine or human serum albumin, for cell culture is a by-product of the physico-chemical, biochemical and cell-specific properties of the molecule. In this review an analysis of these features of albumin leads to a consideration of the extracellular and intracellular actions of the molecule, and importantly the role of its interactions with numerous ligands or bioactive factors that influence the growth of cells in culture: these include hormones, growth factors, lipids, amino acids, metal ions, reactive oxygen and nitrogen species to name a few. The interaction of albumin with the cell in relation to these co-factors has a potential impact on metabolic and biosynthetic activity, cell proliferation and survival. Application of this knowledge to improve the performance in manufacturing biotechnology and in the emerging uses of cell culture for tissue engineering and stem cell derived therapies is an important prospect

Gene Expression Profiling of Iron Deficiency Chlorosis Sensitive and Tolerant Soybean Indicates Key Roles for Phenylpropanoids under Alkalinity Stress

Alkaline soils comprise 30% of the earth and have low plant-available iron (Fe) concentration, and can cause iron deficiency chlorosis (IDC). IDC causes soybean yield losses of $260 million annually. However, it is not known whether molecular responses to IDC are equivalent to responses to low iron supply. IDC tolerant and sensitive soybean lines provide a contrast to identify specific factors associated with IDC.We used RNA-seq to compare gene expression under combinations of normal pH (5.7) or alkaline pH (7.7, imposed by 2.5mM bicarbonate, or pH 8.2 imposed by 5mM bicarbonate) and normal (25μM) or low (1μM) iron conditions from roots of these lines. Thus, we were able to treat pH and Fe supply as separate variables. We also noted differential gene expression between IDC sensitive and tolerant genotypes in each condition. Classical iron uptake genes, including ferric-chelate reductase (FCR) and ferrous transporters, were upregulated by both Fe deficiency and alkaline stress, however, their gene products did not function well at alkaline pH. In addition, genes in the phenylpropanoid synthesis pathway were upregulated in both alkaline and low Fe conditions. These genes lead to the production of fluorescent root exudate (FluRE) compounds, such as coumarins. Fluorescence of nutrient solution increased with alkaline treatment, and was higher in the IDC tolerant line. Some of these genes also localized to previously identified QTL regions associated with IDC. We hypothesize that FluRE become essential at alkaline pH where the classical iron uptake system does not function well. This work could result in new strategies to screen for IDC tolerance, and provide breeding targets to improve crop alkaline stress tolerance

\u3ci\u3ePorocercospora seminalis\u3c/i\u3e gen. et comb. nov., the causal organism of buffalograss false smut

False smut caused by Cercospora seminalis is an important disease of buffalograss (Buchloe dactyloides) affecting seed production. The pathogen prevents normal caryopsis development and causes considerable yield loss and reduced seed germination. The current taxonomic placement of the false-smut causal pathogen in the genus Cercospora is incorrect based on its morphological characteristics and DNA phylogeny. In the present study the phylogenetic position of C. seminalis is clarified based on DNA sequence analysis of three loci namely the internal transcribed spacer (ITS) region, partial nuclear ribosomal large subunit (LSU) and partial sequences of the RNA polymerase II second largest subunit (RPB2). A collection of C. seminalis isolates was made from buffalograss sites near Lincoln, Nebraska. DNA sequence data indicated that Cercospora seminalis is phylogenetically close to but distinct from species of Bipolaris and Curvularia (Pleosporaceae, Pleosporales). Cercospora seminalis morphologically had unique characteristics, namely densely aggregated and repeatedly branched conidiophores arising from a brown stroma, monotretic conidiogenous cells with inconspicuous loci, and scolecosporous conidia with distosepta, and thickened, darkened hila. Porocercospora is introduced as a new genus to accommodate the buffalograss false-smut pathogen

Buffalo Grass Field Data Part 2: Editing and Graphing Data

Build on your skills and learning from Buffalo Grass Experiment Lesson part 1, as we continue to gain experimental skills using buffalo grass plots. Learn how to analyze data by deciphering between good and bad data and knowing when to take out outlying data. Also learn how to manage data by creating a mean and a line graph to make data easier to interpret. Objectives Analyze data to determine which information is authentic and which is a result from error. Identify the importance of combing through data and the impact that it can have on the result. Learn about means and how they are used to manage data. Create a line graph that conveys result of the correct data collected Modules Lesson home Objectives Introduction Analyze data Creating a line graph Line graph assessment Video

Transcriptome profiling of Kentucky bluegrass (\u3ci\u3ePoa pratensis\u3c/i\u3e L.) accessions in response to salt stress

Background: Kentucky bluegrass (Poa pratensis L.) is a prominent turfgrass in the cool-season regions, but it is sensitive to salt stress. Previously, a relatively salt tolerant Kentucky bluegrass accession was identified that maintained green color under consistent salt applications. In this study, a transcriptome study between the tolerant (PI 372742) accession and a salt susceptible (PI 368233) accession was conducted, under control and salt treatments, and in shoot and root tissues. Results: Sample replicates grouped tightly by tissue and treatment, and fewer differentially expressed transcripts were detected in the tolerant PI 372742 samples compared to the susceptible PI 368233 samples, and in root tissues compared to shoot tissues. A de novo assembly resulted in 388,764 transcripts, with 36,587 detected as differentially expressed. Approximately 75 % of transcripts had homology based annotations, with several differences in GO terms enriched between the PI 368233 and PI 372742 samples. Gene expression profiling identified salt-responsive gene families that were consistently down-regulated in PI 372742 and unlikely to contribute to salt tolerance in Kentucky bluegrass. Gene expression profiling also identified sets of transcripts relating to transcription factors, ion and water transport genes, and oxidation-reduction process genes with likely roles in salt tolerance. Conclusions: The transcript assembly represents the first such assembly in the highly polyploidy, facultative apomictic Kentucky bluegrass. The transcripts identified provide genetic information on how this plant responds to and tolerates salt stress in both shoot and root tissues, and can be used for further genetic testing and introgression

Response of \u3ci\u3eAmaranthus\u3c/i\u3e spp. following exposure to sublethal herbicide rates via spray particle drift

The adverse consequences of herbicide drift towards sensitive crops have been extensively reported in the literature. However, little to no information is available on the consequences of herbicide drift onto weed species inhabiting boundaries of agricultural fields. Exposure to herbicide drift could be detrimental to long-term weed management as several weed species have evolved herbicide-resistance after recurrent selection with sublethal herbicide rates This study investigated the deposition of glyphosate, 2,4-D, and dicamba spray particle drift from applications with two different nozzles in a low speed wind tunnel, and their impact on growth and development of Amaranthus spp. Herbicide drift resulted in biomass reduction or complete plant mortality. Inflection points (distance to 50% biomass reduction) for Amaranthus tuberculatus were 7.7, 4.0, and 4.1 m downwind distance for glyphosate, 2,4-D, and dicamba applications with the flat-fan nozzle, respectively, whereas these values corresponded to 2.8, 2.5, and 1.9 m for applications with the air-inclusion nozzle. Inflection points for Amaranthus palmeri biomass reduction were 16.3, 10.9, and 11.5 m for glyphosate, 2,4-D, and dicamba applications with the flat-fan nozzle, respectively, whereas these values corresponded to 7.6, 5.4, and 5.4 m for applications with the air-inclusion nozzle. Plants were more sensitive to glyphosate at higher exposure rates than other herbicides, whereas plants were more sensitive to 2,4-D and dicamba at lower exposure rates compared to glyphosate. Applications with the flat-fan nozzle resulted in 32.3 and 11.5% drift of the applied rate at 1.0 and 3.0 m downwind, respectively, whereas the air-inclusion nozzle decreased the dose exposure in the same distances (11.4 and 2.7%, respectively). Herbicide drift towards field boundaries was influenced by nozzle design and exposed weeds to herbicide rates previously reported to select for herbicide-resistant biotypes