Genome-wide analysis of cold imbibition stress in soybean, Glycine max.

In Canada, the length of the frost-free season necessitates planting crops as early as possible to ensure that the plants have enough time to reach full maturity before they are harvested. Early planting carries inherent risks of cold water imbibition (specifically less than 4°C) affecting seed germination. A marker dataset developed for a previously identified Canadian soybean GWAS panel was leveraged to investigate the effect of cold water imbibition on germination. Seed from a panel of 137 soybean elite cultivars, grown in the field at Ottawa, ON, over three years, were placed on filter paper in petri dishes and allowed to imbibe water for 16 hours at either 4°C or 20°C prior to being transferred to a constant 20°C. Observations on seed germination, defined as the presence of a 1 cm radicle, were done from day two to seven. A three-parameter exponential rise to a maximum equation (3PERM) was fitted to estimate germination, time to the one-half maximum germination, and germination uniformity for each cultivar. Genotype-by-sequencing was used to identify SNPs in 137 soybean lines, and using genome-wide association studies (GWAS - rMVP R package, with GLM, MLM, and FarmCPU as methods), haplotype block analysis, and assumed linkage blocks of ±100 kbp, a threshold for significance was established using the qvalue package in R, and five significant SNPs were identified on chromosomes 1, 3, 4, 6, and 13 for maximum germination after cold water imbibition. Percent of phenotypic variance explained (PVE) and allele substitution effect (ASE) eliminated two of the five candidate SNPs, leaving three QTL regions on chromosomes 3, 6, and 13 (Chr3-3419152, Chr6-5098454, and Chr13-29649544). Based on the gene ontology (GO) enrichment analysis, 14 candidate genes whose function is predicted to include germination and cold tolerance related pathways were identified as candidate genes. The identified QTLs can be used to select future soybean cultivars tolerant to cold water imbibition and mitigate risks associated with early soybean planting.

Long-Term Tillage and Crop Rotation Regimes Reshape Soil-Borne Oomycete Communities in Soybean, Corn, and Wheat Production Systems.

Soil-borne oomycetes include devastating plant pathogens that cause substantial losses in the agricultural sector. To better manage this important group of pathogens, it is critical to understand how they respond to common agricultural practices, such as tillage and crop rotation. Here, a long-term field experiment was established using a split-plot design with tillage as the main plot factor (conventional tillage (CT) vs. no till (NT), two levels) and rotation as the subplot factor (monocultures of soybean, corn, or wheat, and corn-soybean-wheat rotation, four levels). Post-harvest soil oomycete communities were characterized over three consecutive years (2016-2018) by metabarcoding the Internal Transcribed Spacer 1 (ITS1) region. The community contained 292 amplicon sequence variants (ASVs) and was dominated by Globisporangium spp. (85.1% in abundance, 203 ASV) and Pythium spp. (10.4%, 51 ASV). NT decreased diversity and community compositional structure heterogeneity, while crop rotation only affected the community structure under CT. The interaction effects of tillage and rotation on most oomycetes species accentuated the complexity of managing these pathogens. Soil and crop health represented by soybean seedling vitality was lowest in soils under CT cultivating soybean or corn, while the grain yield of the three crops responded differently to tillage and crop rotation regimes.

Variability in Maturity, Oil and Protein Concentration, and Genetic Distinctness among Soybean Accessions Conserved at Plant Gene Resources of Canada.

Soybean (Glycine max (L.) Merr.) is one of the important crops in Canada and has the potential to expand its production further north into the Canadian Prairies. Such expansion, however, requires the search for adapted soybean germplasm useful for the development of productive cultivars with earlier maturity and increased protein concentration. We initiated several research activities to characterize 848 accessions of the soybean collection conserved at Plant Gene Resources of Canada (PGRC) for maturity, oil and protein concentration, and genetic distinctness. The characterization revealed a wide range of variations present in each assessed trait among the PGRC soybean accessions. The trait variabilities allowed for the identification of four core subsets of 35 PGRC soybean accessions, each specifically targeted for early maturity for growing in Saskatoon and Ottawa, and for high oil and protein concentration. The two early maturity core subsets for Saskatoon and Ottawa displayed days to maturity ranging from 103 to 126 days and 94 to 102 days, respectively. The two core subsets for high oil and protein concentration showed the highest oil and protein concentration from 25.0 to 22.7% and from 52.8 to 46.7%, respectively. However, these core subsets did not differ significantly in genetic distinctness (as measured with 19,898 SNP markers across 20 soybean chromosomes) from the whole PGRC soybean collection. These findings are useful, particularly for the management and utilization of the conserved soybean germplasm.

Novel QTL for Low Seed Cadmium Accumulation in Soybean.

Soybean is a valuable crop, used in animal feed and for human consumption. Selecting soybean cultivars with low seed cadmium (Cd) concentration is important for the purpose of minimizing the transfer of Cd into the human body. To ensure international trade, farmers need to produce soybean that meets the European Union (EU) Cd limit of 0.2 mg kg-1. In this study, we evaluated two populations of recombinant inbred lines (RILs), X5154 and X4050, for seed Cd accumulation. Linkage maps were constructed with 325 and 280 polymorphic simple sequence repeat (SSR) markers, respectively, and used to identify a novel minor quantitative trait locus (QTL) on chromosome 13 in the X4050 population between SSR markers Satt522 and Satt218. Based on a gene ontology search within the QTL region, seven genes were identified as candidates responsible for low seed Cd accumulation, including Glyma.13G308700 and Glyma.13G309100. In addition, we confirmed the known major gene, Cda1, in the X5154 population and developed KASP and CAPS/dCAPS allele-specific markers for efficient marker-assisted breeding for Cda1.

Photoperiod Affects Node Appearance Rate and Flowering in Early Maturing Soybean.

The photoperiod plays a critical role in the control of flowering timing in soybean (Glycine max (L.) Merr.) with long days increasing the time to flowering. Early flowering cultivars have been developed from breeding programs for environments with long photoperiods; however, this effect is challenging to isolate in field experiments because of other environmental influences. Our experiment examined the effect of photoperiod on the node appearance rate and time to flower for 13 early maturing soybean cultivars ranging in maturity group (MG) between 000.9 and 1.3. Growth chambers were programmed to 14, 15, 16, and 17 h photoperiods and temperature was kept at 25 °C. The date of emergence and main stem node appearance were recorded until flowering. The node appearance rate was slowest for the first node and increased thereafter. All cultivars required more time to flowering in the longer photoperiod treatments and the later rated MG had the greatest sensitivity to photoperiod. A delay in time to flower from a longer photoperiod can delay maturity and expose the crop to fall frost that can reduce seed yield and quality. Understanding and documentation of soybean photoperiod sensitivity will help plant breeders develop suitable cultivars for environments with long photoperiods.

Patterns of Genetic Variation in a Soybean Germplasm Collection as Characterized with Genotyping-by-Sequencing.

Genomic characterization is playing an increasing role in plant germplasm conservation and utilization, as it can provide higher resolution with genome-wide SNP markers than before to identify and analyze genetic variation. A genotyping-by-sequencing technique was applied to genotype 541 soybean accessions conserved at Plant Gene Resources of Canada and 30 soybean cultivars and breeding lines developed by the Ottawa soybean breeding program of Agriculture and Agri-Food Canada. The sequencing generated an average of 952,074 raw sequence reads per sample. SNP calling identified 43,891 SNPs across 20 soybean chromosomes and 69 scaffolds with variable levels of missing values. Based on 19,898 SNPs with up to 50% missing values, three distinct genetic groups were found in the assayed samples. These groups were a mixture of the samples that originated from different countries and the samples of known maturity groups. The samples that originated from Canada were clustered into all three distinct groups, but 30 Ottawa breeding lines fell into two groups only. Based on the average pairwise dissimilarity estimates, 40 samples with the most genetic distinctness were identified from three genetic groups with diverse sample origin and known maturity. Additionally, 40 samples with the highest genetic redundancy were detected and they consisted of different sample origins and maturity groups, largely from one genetic group. Moreover, some genetically duplicated samples were identified, but the overall level of genetic duplication was relatively low in the collection. These findings are useful for soybean germplasm management and utilization.

Genome-wide association identifies several QTLs controlling cysteine and methionine content in soybean seed including some promising candidate genes.

Soybean is an important source of protein, oil and carbohydrates, as well as other beneficial nutrients. A major function of proteins in nutrition is to supply adequate amounts of amino acids. Although they are essential for human nutrition, the sulfur-containing amino acids cysteine (Cys) and methionine (Met) are often limited and the genetic control of their content in soybean seeds is poorly characterized. This study aimed to characterize the phenotypic variation and identify quantitative trait loci (QTL) associated with Cys and Met content in a core set of 137 soybean lines, representative of the genetic diversity among Canadian short-season soybean, spanning maturity groups 000-II (MG000-II). Significant phenotypic differences were found among these lines for Cys, Met and Cys + Met content. Using both a mixed linear model and six multi-locus methods with a catalogue of 2.18 M SNPs, we report a total of nine QTLs and seventeen QTNs of which seven comprise promising candidate genes. This work allowed us to reproducibly detect multiple novel loci associated with sulfur-containing amino acid content. The markers and genes identified in this study may be useful for soybean genetic improvement aiming to increase Cys and Met content.

Identification of loci controlling mineral element concentration in soybean seeds.

BACKGROUND: Mineral nutrients play a crucial role in the biochemical and physiological functions of biological systems. The enhancement of seed mineral content via genetic improvement is considered as the most promising and cost-effective approach compared alternative means for meeting the dietary needs. The overall objective of this study was to perform a GWAS of mineral content (Ca, K, P and S) in seeds of a core set of 137 soybean lines that are representative of the diversity of early maturing soybeans cultivated in Canada (maturity groups 000-II). RESULTS: This panel of 137 soybean lines was grown in five environments (in total) and the seed mineral content was measured using a portable x-ray fluorescence (XRF) spectrometer. The association analyses were carried out using three statistical models and a set of 2.2 million SNPs obtained from a combined dataset of genotyping-by-sequencing and whole-genome sequencing. Eight QTLs significantly associated with the Ca, K, P and S content were identified by at least two of the three statistical models used (in two environments) contributing each from 17 to 31% of the phenotypic variation. A strong reproducibility of the effect of seven out these eight QTLs was observed in three other environments. In total, three candidate genes were identified involved in transport and assimilation of these mineral elements. CONCLUSIONS: There have been very few GWAS studies to identify QTLs associated with the mineral element content of soybean seeds. In addition to being new, the QTLs identified in this study and candidate genes will be useful for the genetic improvement of soybean nutritional quality through marker-assisted selection. Moreover, this study also provides details on the range of phenotypic variation encountered within the Canadian soybean germplasm.

Genetic Analysis of High Protein Content in 'AC Proteus' Related Soybean Populations Using SSR, SNP, DArT and DArTseq Markers.

Key message: Several AC Proteus derived genomic regions (QTLs, SNPs) have been identified which may prove useful for further development of high yielding high protein cultivars and allele-specific marker developments. High seed protein content is a trait which is typically difficult to introgress into soybean without an accompanying reduction in seed yield. In a previous study, 'AC Proteus' was used as a high protein source and was found to produce populations that did not exhibit the typical association between high protein and low yield. Five high x low protein RIL populations and a high x high protein RIL population were evaluated by either quantitative trait locus (QTL) analysis or bulk segregant analyses (BSA) following phenotyping in the field. QTL analysis in one population using SSR, DArT and DArTseq markers found two QTLs for seed protein content on chromosomes 15 and 20. The BSA analyses suggested multiple genomic regions are involved with high protein content across the five populations, including the two previously mentioned QTLs. In an alternative approach to identify high protein genes, pedigree analysis identified SNPs for which the allele associated with high protein was retained in seven high protein descendants of AC Proteus on chromosomes 2, 17 and 18. Aside from the two identified QTLs (five genomic regions in total considering the two with highly elevated test statistic, but below the statistical threshold and the one with epistatic interactions) which were some distance from Meta-QTL regions and which were also supported by our BSA analysis within five populations. These high protein regions may prove useful for further development of high yielding high protein cultivars.

Soybean Yield and Seed Composition Changes in Response to Increasing Atmospheric CO2 Concentration in Short-Season Canada.

From 1993, we have conducted trials with the same set of old to newer soybean cultivars to determine the impact of plant breeding on seed yield, physiological and agronomic characteristics, and seed composition. Since 1993, global atmospheric [CO2] increased by 47 ppm. The objective of our current analysis with this data set was to determine if there were changes in soybean seed yield, quality or phenology attributable to elevated atmospheric CO2 concentration (eCO2), temperature or precipitation. Additionally, we estimated genetic gain annually. Over 23 years, there was a significant increase in atmospheric [CO2] but not in-season average maximum or minimum temperatures, or average in-season precipitation. Seed yield was increased significantly by eCO2, higher precipitation and higher minimum temperatures during flowering and podding. Yield decreased with higher minimum temperatures during vegetative growth and seed filling. Seed oil and also seed protein plus oil concentrations were both reduced with eCO2. Phenology has also changed, with soybean cultivars spending less time in vegetative growth, while time to maturity remained constant. Over the 23 years of the study, genetic improvement rates decreased as [CO2] increased. Newer cultivars are not better adapted to eCO2 and soybean breeders may need to intentionally select for favourable responses to eCO2 in the future.

Optimal nonlinear filtering using the finite-volume method.

Optimal sequential inference, or filtering, for the state of a deterministic dynamical system requires simulation of the Frobenius-Perron operator, that can be formulated as the solution of a continuity equation. For low-dimensional, smooth systems, the finite-volume numerical method provides a solution that conserves probability and gives estimates that converge to the optimal continuous-time values, while a Courant-Friedrichs-Lewy-type condition assures that intermediate discretized solutions remain positive density functions. This method is demonstrated in an example of nonlinear filtering for the state of a simple pendulum, with comparison to results using the unscented Kalman filter, and for a case where rank-deficient observations lead to multimodal probability distributions.

Quantifying the Effects of Photoperiod, Temperature and Daily Irradiance on Flowering Time of Soybean Isolines.

Soybean isolines with different combinations of photoperiod sensitivity alleles were planted in a greenhouse at different times during the year resulting in natural variation in daily incident irradiance and duration. The time from planting to first flower were observed. Mathematical models, using additive and multiplicative modes, were developed to quantify the effect of photoperiod, temperature, photoperiod-temperature interactions, rate of photoperiod change, and daily solar irradiance on flowering time. Observed flowering times correlated with predicted times (R² = 0.92, Standard Error of the Estimate (SSE) = 2.84 d, multiplicative mode; R² = 0.91, SSE = 2.88 d, additive mode). The addition of a rate of photoperiod change function and an irradiance function to the temperature and photoperiod functions improved the accuracy of flowering time prediction. The addition of a modified photoperiod function, which allowed for photoperiod sensitivity at shorter photoperiods, improved prediction of flowering time. Both increasing and decreasing rate of photoperiod change, as well as low levels of daily irradiance delayed flowering in soybean. The complete model, which included terms for the rate of photoperiod change, photoperiod, temperature and irradiance, predicted time to first flower in soybean across a range of environmental conditions with an SEE of 3.6 days when tested with independent data.

Mapping and validation of simple sequence repeat markers linked to a major gene controlling seed cadmium accumulation in soybean [Glycine max (L.) Merr].

Daily consumption of cadmium (Cd) contaminated foods poses a risk to human health. Cultivar selection is an important method to limit Cd uptake and accumulation; however, analyzing grain Cd concentration is costly and time-consuming. Developing markers for low Cd accumulation will facilitate marker assisted selection (MAS). Inheritance studies using a threshold value of 0.2 mg kg(-1) for low and high and an F(2:3) population showed that low Cd accumulation in soybean seed is under the control of a major gene (Cda1, proposed name) with the allele for low accumulation being dominant. A recombinant inbred line (RIL) population (F(6:8)) derived from the cross AC Hime (high Cd accumulation) and Westag-97 (low Cd accumulation) was used to identify the DNA markers linked to Cda gene(s) or quantitative trait loci (QTLs) controlling low Cd accumulation. We screened 171 simple sequence repeat (SSR) primers that showed polymorphism between parents on the 166 RILs. Of these, 40 primers were newly developed from the soybean genomic DNA sequence. Seven SSR markers, SatK138, SatK139, SatK140 (0.5 cM), SatK147, SacK149, SaatK150 and SattK152 (0.3 cM), were linked to Cda1 in soybean seed. All the linked markers were mapped to the same linkage group (LG) K. The closest flanking SSR markers linked to Cda1 were validated using a parallel population (RILs) involving Leo x Westag-97. Linked markers were also validated with diverse soybean genotypes differing in their seed Cd concentration and showed that SSR markers SatK147, SacK149, and SattK152 clearly differentiated the high and low Cd accumulating genotypes tested. To treat Cd uptake as a quantitative trait, QTL analysis using a linkage map constructed with 161 markers identified a major QTL associated with low Cd concentration in the seeds. The QTL was also mapped to the same location as Cda1 on LG-K. This QTL accounted for 57.3% of the phenotypic variation. Potential candidate genes (genes with known or predicted function that could influence the seed Cd concentration) like protein kinase, putative Adagio-like protein, and plasma membrane H(+)-ATPase were found to be located in the locus of interest. Of the four SSR markers located in the region, SattK152 was localized in the plasma membrane H(+)-ATPase gene. SSR markers closely linked to Cda1 in seeds of soybean were identified and have potential to be used for MAS to develop low Cd accumulating cultivars in a breeding program.

Regulation of seed yield and agronomic characters by photoperiod sensitivity and growth habit genes in soybean.

Soybean genotypes are adapted to narrow bands of latitude due to photoperiod sensitivity. There are several photoperiod-sensitive loci (E1, E2, E3, E4, E5, E6, E7, E8). Determinate and indeterminate growth habits are controlled by a single locus. The objective of our research was to examine the effects of photoperiod sensitivity and growth habit alleles on seed yield and other agronomic characters using isogenic lines. Twenty 'Harosoy' isolines with 11 photoperiod-sensitive genotypes many with both indeterminate and determinate growth habits were grown in the field at Ottawa, ON, from 2003 to 2007. Maturity ranged from 97 to 127 days, and seed yield increased linearly with maturity until about 112 days when it plateaued. Determinate lines were always shorter than indeterminate lines of equivalent maturity. Seed yield was associated with plant height, maturity, seed sugar concentration, seed weight and lodging. Effects of alleles at individual loci, and additive and epistatic effects across multiple loci were examined. At a single locus, photoperiod-insensitive alleles produced isolines that matured 8-11 days earlier, yielded less, and had shorter plants with reduced lodging. In multiple loci analyses, additive effects explained most of the variation in agronomic characters since additive models with E1, E3, E4, E7 and Dt1 loci included compared well to additive plus epistatic models and genotype-based models. Variation in photoperiod sensitivity and growth habit alleles results in a range of maturity, with pleiotropic effects on seed yield and agronomic characteristics, and play an important role in providing adaptation across latitudes.

Sir Charles Edward Saunders, Dominion cerealist.

Charles Edward Saunders was born in London, Ontario, in 1867. His father, Sir William Saunders, was the first director of the Dominion Experimental Farms (1886-1911). Charles received his B.A. with honours in science from the University of Toronto in 1888 and his Ph.D. in chemistry from Johns Hopkins University in Baltimore, Maryland, in 1891. He attempted a career in music, his first love, from 1893 to 1902. With his father, Charles attended the 1902 International Conference on Plant Breeding and Hybridization in New York, where he learned of Mendel's theories of inheritance and their applicability to plant breeding. When he began work in 1903 in the Division of Cereal Breeding and Experimentation at the Central Experimental Farm in Ottawa, he used the knowledge he had gained at that conference. It was Charles's goal to achieve "fixity" in the varieties that had been bred and released using phenotypic mass selection, prior to his tenure as Cerealist. He selected four heads from the wheat variety Markham and in the winter of 1904 he performed a "chewing test" to select for gluten elasticity and colour. Seeds from two heads were chosen, and seeds from one went on to produce the variety Marquis after extensive yield trials on the Prairies. Marquis was 7 to 10 days earlier than Red Fife, the standard bread wheat of the Prairies. The earliness and tremendous yield of Marquis wheat resulted in the rapid and successful settlement of the Great Plains and countless billions of dollars in revenue to Canada. By 1923, 90% of the spring wheat in Canada and 70% in the USA was Marquis. Charles continued as Dominion Cerealist until his retirement in 1922. He was knighted in 1934, and died in 1937.

Nurse educators are key to a best practices implementation program.

In a statewide Best Practices Project, the involvement of nurse educators in the implementation process improved the overall success of participating long-term care facilities in improving the quality of resident care. Two nurse educators provided consultation and training to nursing facility staff on three best practice protocols: prevention of decline in the activities of daily living of eating and dressing, pain, and depression. Facility staff were given protocols to follow, together with documentation forms to assist in determining resident appropriateness for participation in the assigned best practice, assessing resident involvement in the protocol over time, and tracking staff progress with the implementation. The success of the 2-year first phase of the project in significantly improving measurable resident quality of care in best practice protocol areas has resulted in a second phase involving additional facilities and protocols.

Outcome of asymptomatic electric shock victims requesting an emergency ambulance.

OBJECTIVE: To determine whether patients suffering from electric shock without significant symptoms at the point of an emergency call could be appropriately assigned a nonemergency ambulance response using the Medical Priority Dispatch System (MPDS). METHODS: Welsh Ambulance Service dispatch records were searched to identify patients allocated the MPDS code of 15C01 (electric shock without priority symptoms) over a 30-month period. Ambulance and hospital records were also reviewed. RESULTS: Records were unavailable for seven of the 52 patients identified (13%). Nine refused transport (17%, 95% CI 8.2% to 30.3%), and 36 were taken to emergency departments (69%, 95% CI 55% to 81%). Of the 52 patients, 23 were discharged shortly after arrival (44%, 95% CI 31% to 59%), 11 receiving no treatment (21%, 95% CI 11% to 35%). Thirteen patients were admitted as inpatients or were transferred to another hospital (25% of the total sample, 95% CI 14% to 39%). Fourteen had burns (27% of the total sample, 95% CI 16% to 41%). Five of these were admitted or transferred to another hospital, five were discharged after treatment, and two left without treatment (one self-discharged). Records were unavailable for two burns patients. CONCLUSIONS: The findings of this study do not justify allocating a low-priority response to victims of electric shock without significant symptoms at the point of the emergency call, since 25% require hospital admission. Further research is required to determine whether the addition of questions to the MPDS about burns and pregnancy might allow safe allocation of a nonemergency response to other asymptomatic electric shock patients.