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Nutrient requirements for para-athletes will be influenced by a variety of factors secondary to their impairment and, therefore, recommendations for para-athletes cannot be drawn directly from that of able-bodied athletes. Information on the dietary intakes of para-athletes is lacking and therefore needs to be examined. This study assessed the nutrient intakes and diet quality of 31 paracyclists via food frequency questionnaires. Based on the dietary reference intakes, most paracyclists consumed intakes above the recommended dietary allowance (RDA) or adequate intake (AI). Recommendations were not met for iodine (males = 87% RDA; females = 62% RDA) or fibre in males (84% AI). A 26% risk of inadequacy was noted for vitamins D and E in females. A total of 42% of females and 75% of males did not meet fibre recommendations (14 g/1000 kcal), and only three athletes (all females) consumed fatty acids in the recommended omega-6 to omega-3 ratio of 4:1 or less. Athletes consumed grains, fruits, and vegetables frequently, though whole grains, pulses, and oily fish were generally consumed less often. Paracyclists appear to be consuming intakes at or above recommendations for most nutrients, though several nutrients were consumed below the recommended amounts; therefore, increasing the variety of foods consumed is suggested.

Monocarpic plants have a single reproductive phase in their life. Therefore, flower and fruit production are restricted to the length of this period. This reproductive strategy involves the regulation of flowering cessation by a coordinated arrest of the growth of the inflorescence meristems, optimizing resource allocation to ensure seed filling. Flowering cessation appears to be a regulated phenomenon in all monocarpic plants. Early studies in several species identified seed production as a major factor triggering inflorescence proliferative arrest. Recently, genetic factors controlling inflorescence arrest, in parallel to the putative signals elicited by seed production, have started to be uncovered in Arabidopsis, with the MADS-box gene FRUITFULL (FUL) playing a central role in the process. However, whether the genetic network regulating arrest is also at play in other species is completely unknown. Here, we show that this role of FUL is not restricted to Arabidopsis but is conserved in another monocarpic species with a different inflorescence structure, field pea, strongly suggesting that the network controlling the end of flowering is common to other plants. Moreover, field trials with lines carrying mutations in pea FUL genes show that they could be used to boost crop yield., Competing Interests: Competing interests statement:The authors declare no competing interest.

Improving the seed protein concentration (SPC) of pea ( Pisum sativum L.) has turned into an important breeding objective because of the consumer demand for plant-based protein and demand from protein fractionation industries. To support the marker-assisted selection (MAS) of SPC towards accelerated breeding of improved cultivars, we have explored two diverse recombinant inbred line (RIL) populations to identify the quantitative trait loci (QTLs) associated with SPC. The two RIL populations, MP 1918 × P0540-91 (PR-30) and Ballet × Cameor (PR-31), were derived from crosses between moderate SPC × high SPC accessions. A total of 166 and 159 RILs of PR-30 and PR-31, respectively, were genotyped using an Axiom® 90K SNP array and 13.2K SNP arrays, respectively. The RILs were phenotyped in replicated trials in two and three locations of Saskatchewan, Canada in 2020 and 2021, respectively, for agronomic assessment and SPC. Using composite interval mapping, we identified three QTLs associated with SPC in PR-30 and five QTLs in PR-31, with the LOD value ranging from 3.0 to 11.0. A majority of these QTLs were unique to these populations compared to the previously known QTLs for SPC. The QTL SPC-Ps-5.1 overlapped with the earlier reported SPC associated QTL PC-QTL-3. Three QTLs, SPC-Ps-4.2 , SPC-Ps-5.1 , and SPC-Ps-7.2 with LOD scores of 7.2, 7.9, and 11.3, and which explained 14.5%, 11.6%, and 11.3% of the phenotypic variance, respectively, can be used for marker-assisted breeding to increase SPC in peas. Eight QTLs associated with the grain yield were identified with LOD scores ranging from 3.1 to 8.2. Two sets of QTLs, SPC-Ps-2.1 and GY-Ps-2.1 , and SPC-Ps-5.1 and GY-Ps-5.3 , shared the QTL/peak regions. Each set of QTLs contributed to either SPC or grain yield depending on which parent the QTL region is derived from, thus confirming that breeding for SPC should take into consideration the effects on grain yield., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Gali, Jha, Tar’an, Burstin, Aubert, Bing, Arganosa and Warkentin.)

Heat-moisture treatment (HMT) was employed to modify wrinkled pea (74.2 % and 76.5 % amylose) and round pea starches (35.9 % and 34.8 % amylose) at 35.0 % moisture, 110 or 130 °C, and 6 h. HMT increased the gelatinization temperatures and decreased the gelatinization enthalpy changes, reduced the pasting viscosities and gel hardness, and enhanced the enzymatic resistance of the pea starches in comparison with the native counterparts, with greater extents of changes observed for HMT at 130 °C overall. Although HMT decreased the relative crystallinity and elevated the proportion of amorphous conformation, the remaining double-helical crystallites in the modified samples showed improved thermal stability as revealed by differential scanning calorimetry (DSC). More importantly, the HMT-modified pea starches required a higher heating temperature of 120 °C, rather than 95 °C, in Rapid Visco Analyser to provide greater pasting viscosities and develop firmer gels, suggesting that the modified samples had stronger molecular entanglement than the native counterparts. Such molecular entanglement could also reduce enzymatic digestion of HMT-modified starches after boiling in water. With more diverse functional profiles and increased resistant starch (RS) contents (particularly for the HMT-modified wrinkled pea starches having 22.7-29.9 % RS), the HMT-modified pea starches could be promising new ingredients for food applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Dry pea ( Pisum sativum ) seeds are valuable sources of plant protein, dietary fiber, and starch, but their uses in food products are restricted to some extent due to several off-flavor compounds. Saponins are glycosylated triterpenoids and are a major source of bitter, astringent, and metallic off-flavors in pea products. β-amyrin synthase ( BAS ) is the entry point enzyme for saponin biosynthesis in pea and therefore is an ideal target for knock-out using CRISPR/Cas9 genome editing to produce saponin deficient pea varieties. Here, in an elite yellow pea cultivar (CDC Inca), LC/MS analysis identified embryo tissue, not seed coat, as the main location of saponin storage in pea seeds. Differential expression analysis determined that PsBAS1 was preferentially expressed in embryo tissue relative to seed coat and was selected for CRISPR/Cas9 genome editing. The efficiency of CRISPR/Cas9 genome editing of PsBAS1 was systematically optimized in pea hairy roots. From these optimization procedures, the AtU6-26 promoter was found to be superior to the CaMV35S promoter for gRNA expression, and the use of 37°C was determined to increase the efficiency of CRISPR/Cas9 genome editing. These promoter and culture conditions were then applied to stable transformations. As a result, a bi-allelic mutation (deletion and inversion mutations) was generated in the PsBAS1 coding sequence in a T 1 plant, and the segregated psbas1 plants from the T 2 population showed a 99.8% reduction of saponins in their seeds. Interestingly, a small but statistically significant increase (~12%) in protein content with a slight decrease (~5%) in starch content was observed in the psbas1 mutants under phytotron growth conditions. This work demonstrated that flavor-improved traits can be readily introduced in any pea cultivar of interest using CRISPR/Cas9. Further field trials and sensory tests for improved flavor are necessary to assess the practical implications of the saponin-free pea seeds in food applications., Competing Interests: The authors did not report any conflict of interest., (© 2024 The Authors. Plant Direct published by American Society of Plant Biologists and the Society for Experimental Biology and John Wiley & Sons Ltd.)

Soybean is an important global source of plant-based protein. A persistent trend has been observed over the past two decades that soybeans grown in western Canada have lower seed protein content than soybeans grown in eastern Canada. In this study, 10 soybean genotypes ranging in average seed protein content were grown in an eastern location (control) and three western locations (experimental) in Canada. Seed protein and oil contents were measured for all lines in each location. RNA-sequencing and differential gene expression analysis were used to identify differentially expressed genes that may account for relatively low protein content in western-grown soybeans. Differentially expressed genes were enriched for ontologies and pathways that included amino acid biosynthesis, circadian rhythm, starch metabolism, and lipid biosynthesis. Gene ontology, pathway mapping, and quantitative trait locus (QTL) mapping collectively provide a close inspection of mechanisms influencing nitrogen assimilation and amino acid biosynthesis between soybeans grown in the East and West. It was found that western-grown soybeans had persistent upregulation of asparaginase (an asparagine hydrolase) and persistent downregulation of asparagine synthetase across 30 individual differential expression datasets. This specific difference in asparagine metabolism between growing environments is almost certainly related to the observed differences in seed protein content because of the positive correlation between seed protein content at maturity and free asparagine in the developing seed. These results provided pointed information on seed protein-related genes influenced by environment. This information is valuable for breeding programs and genetic engineering of geographically optimized soybeans., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Hooker, Smith, Zapata, Charette, Luckert, Mohr, Daba, Warkentin, Hadinezhad, Barlow, Hou, Lefebvre, Golshani, Cober and Samanfar.)

Background: Iron deficiency (ID) is the most prevalent micronutrient deficiency in the world and the leading cause of anemia globally. Female athletes are at a disproportionate risk for ID due to blood loss through menstruation and decreased iron absorption secondary to exercise. Field peas are a rich source of iron but, similar to iron from other plant-based sources, the iron has limited bioavailability due to high levels of phytic acid, an inherent compound that binds to cations, creating a salt (phytate), which limits absorption during digestion. Aim: The purpose of our research was to investigate the effect of a field pea variety bred to have low levels of phytic acid on plasma ferritin, exercise performance, and body composition in female runners. Methods: Twenty-eight female runners (age:34.6  ±  9.7 years; weight: 65.1  ±  8.1 kg; VO 2 max: 50.7  ±  8.9 ml/kg/min) underwent measures of ferritin, exercise performance, and body composition before and after being randomly assigned to consume a powder derived from regular peas, low phytic acid peas, or a non-pea control (maltodextrin), plus vitamin C for 8 weeks. Results: The regular pea and low phytic acid pea groups had a 14.4% and 5.1% increase in plasma ferritin, respectively, while the maltodextrin group had a decrease of 2.2%; however, the difference in changes between groups was not statistically significant. No differences between groups were evident in any of the other measures. Conclusion: Larger doses or longer duration of pea supplementation may be necessary to induce meaningful changes in iron status. This trial was registered at ClinicalTrials.gov (NCT04872140).

With the expanding interest in plant-based proteins in the food industry, increasing emphasis is being placed on breeding for protein concentration and quality. Two protein quality traits i.e., amino acid profile and protein digestibility, were assessed in replicated, multi-location field trials from 2019 to 2021 in pea recombinant inbred line population PR-25. This RIL population was targeted specifically for the research of protein related traits and its parents, CDC Amarillo and CDC Limerick, had distinct variations in the concentration of several amino acids. Amino acid profile was determined using near infrared reflectance analysis, and protein digestibility was through an in vitro method. Several essential amino acids were selected for QTL analysis, including lysine, one of the most abundant essential amino acids in pea, and methionine, cysteine, and tryptophan, the limiting amino acids in pea. Based on phenotypic data of amino acid profiles and in vitro protein digestibility of PR-25 harvested in seven location-years, three QTLs were associated with methionine + cysteine concentration, among which, one was located on chromosome 2 (R 2 = 17%, indicates this QTL explained 17% phenotypic variation of methionine + cysteine concentration within PR-25), and two were located on chromosome 5 (R 2 = 11% and 16%). Four QTLs were associated with tryptophan concentration and are located on chromosome 1 (R 2 = 9%), chromosome 3 (R 2 = 9%), and chromosome 5 (R 2 = 8% and 13%). Three QTLs were associated with lysine concentration, among which, one was located on chromosome 3 (R 2 = 10%), the other two were located on chromosome 4 (R 2 = 15% and 21%). Two QTLs were associated with in vitro protein digestibility, one each located on chromosomes 1 (R 2 = 11%) and 2 (R 2 = 10%). QTLs associated with in vitro protein digestibility, and methionine + cysteine concentration on chromosome 2 were identified to be co-localized with known QTL for total seed protein concentration in PR-25. QTLs associated with tryptophan and methionine + cysteine concentration co-localized on chromosome 5. The identification of QTLs associated with pea seed quality is an important step towards marker-assisted selection of breeding lines with improved nutritional quality, which will further boost the competitiveness of pea in plant-based protein markets., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Zhou, Wan, Gali, Jha, Nickerson, House, Tar’an and Warkentin.)

Vegetal diamine oxidase (vDAO), an enzyme proposed to relieve symptoms of histaminosis, shows better reactivity with histamine and aliphatic diamines, as well as higher enzymatic activity than DAO of animal origin. The objective of this study was to evaluate the enzyme activity of vDAO from germinating grains from Lathyrus sativus (grass pea) and Pisum sativum (pea), and to verify the presence of a neurotoxin, β-N-Oxalyl-L-α,β-diaminopropionic acid (β-ODAP), in the crude extract obtained from their seedlings. A targeted liquid chromatography-multiple-reaction monitoring mass spectrometry method was developed and used to quantify β-ODAP in the analysed extracts. An optimized sample preparation procedure, involving protein precipitation with acetonitrile followed by mixed-anion exchange solid-phase extraction, allowed for high sensitivity and good peak shape for β-ODAP detection. The Lathyrus sativus extract exhibited the highest vDAO enzyme activity of the extracts, followed by the extract from pea cultivar Amarillo from the Crop Development Centre (CDC). The results have also shown that even though β-ODAP was present in the crude extract from L. sativus , its content was far below the toxicity threshold (300 mg of β-ODAP/kg body/day). CDC Amarillo showed 5000-fold less β-ODAP than the undialysed L. sativus extract. It was concluded that both species can be considered as convenient sources of vDAO for potential therapeutic use.

Soybean ( Glycine max (L.) Merr.) is among the most valuable crops based on its nutritious seed protein and oil. Protein quality, evaluated as the ratio of glycinin (11S) to β-conglycinin (7S), can play a role in food and feed quality. To help uncover the underlying differences between high and low protein soybean varieties, we performed differential expression analysis on high and low total protein soybean varieties and high and low 11S soybean varieties grown in four locations across Eastern and Western Canada over three years (2018-2020). Simultaneously, ten individual differential expression datasets for high vs. low total protein soybeans and ten individual differential expression datasets for high vs. low 11S soybeans were assessed, for a total of 20 datasets. The top 15 most upregulated and the 15 most downregulated genes were extracted from each differential expression dataset and cross-examination was conducted to create shortlists of the most consistently differentially expressed genes. Shortlisted genes were assessed for gene ontology to gain a global appreciation of the commonly differentially expressed genes. Genes with roles in the lipid metabolic pathway and carbohydrate metabolic pathway were differentially expressed in high total protein and high 11S soybeans in comparison to their low total protein and low 11S counterparts. Expression differences were consistent between East and West locations with the exception of one, Glyma.03G054100 . These data are important for uncovering the genes and biological pathways responsible for the difference in seed protein between high and low total protein or 11S cultivars.

Over the past two decades soybeans grown in western Canada have persistently had lower seed protein than those grown in eastern Canada. To understand the discrepancy in seed protein content between eastern- and western-grown soybeans, RNA-seq and differential expression analysis have been investigated. Ten soybean genotypes, ranging from low to high in seed protein content, were grown in four locations across eastern (Ottawa) and western (Morden, Brandon, and Saskatoon) Canada. Differential expression analysis revealed 34 differentially expressed genes encoding Glycine max Sugars Will Eventually be Exported Transporters (GmSWEETs), including paralogs GmSWEET29 and GmSWEET34 (AtSWEET2 homologs) that were consistently upregulated across all ten genotypes in each of the western locations over three years. GmSWEET29 and GmSWEET34 are likely candidates underlying the lower seed protein content of western soybeans. GmSWEET20 (AtSWEET12 homolog) was downregulated in the western locations and may also play a role in lower seed protein content. These findings are valuable for improving soybean agriculture in western growing regions, establishing more strategic and efficient agricultural practices., Competing Interests: The authors declare no conflict of interest.

This research aimed to identify quantitative trait loci (QTLs) associated with seed protein concentration in a recombinant inbred line (RIL) population of pea and aimed to validate the identified QTLs using chromosome segment-introgressed lines developed by recurrent backcrossing. PR-25, an RIL population consisting of 108 F7 bulked lines derived from a cross between CDC Amarillo (yellow cotyledon) and CDC Limerick (green cotyledon), was used in this research. The RIL population was genotyped using an Axiom 90K SNP array. A total of 10,553 polymorphic markers were used for linkage map construction, after filtering for segregation distortion and missing values. The linkage map represents 901 unique loci on 11 linkage groups which covered a map distance of 855.3 Centimorgans. Protein concentration was assessed using near-infrared (NIR) spectroscopy of seeds harvested from field trials in seven station-years in Saskatchewan, Canada, during the 2019-2021 field seasons. Three QTLs located on chromosomes 2, 3 and 5 were identified to be associated with seed protein concentration. These QTLs explained 22%, 11% and 17% of the variation for protein concentration, respectively. The identified QTLs were validated by introgression lines, developed by marker-assisted selection of backcross lines for introgression of corresponding chromosome segments (~1/4 chromosome) harboring the QTL regions. Introgression line PR-28-7, not carrying any protein-related QTLs identified in this study, was 4.7% lower in protein concentration than CDC Amarillo, the lower protein parent of PR-25 which carried one identified protein-related QTL. The SNP markers located at the peak of the three identified QTLs will be converted into breeder-friendly KASP assays, which will be used for the selection of high-protein lines from segregating populations.

Individuals may opt to follow a plant-based diet for a variety of reasons, such as religious practices, health benefits or concerns for animal or environmental welfare. Such diets offer a broad spectrum of health benefits including aiding in the prevention and management of chronic diseases. In addition to health benefits, a plant-based diet may provide performance-enhancing effects for various types of exercise due to high carbohydrate levels and the high concentration of antioxidants and phytochemicals found in a plant-based diet. However, some plant-based foods also contain anti-nutrional factors, such as phytate and tannins, which decrease the bioavailability of key nutrients, such as iron, zinc, and protein. Thus, plant-based diets must be carefully planned to ensure adequate intake and absorption of energy and all essential nutrients. The current narrative review summarizes the current state of the research concerning the implications of a plant-based diet for health and exercise performance. It also outlines strategies to enhance the bioavailability of nutrients, sources of hard-to-get nutrients, and sport supplements that could interest plant-based athletes., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Heat and drought, individually or in combination, limit pea productivity. Fortunately, substantial genetic diversity exists in pea germplasm for traits related to abiotic stress resistance. Understanding the genetic basis of resistance could accelerate the development of stress-adaptive cultivars. We conducted a genome-wide association study (GWAS) in pea on six stress-adaptive traits with the aim to detect the genetic regions controlling these traits. One hundred and thirty-five genetically diverse pea accessions were phenotyped in field studies across three or five environments under stress and control conditions. To determine marker trait associations (MTAs), a total of 16,877 valuable single nucleotide polymorphisms (SNPs) were used in association analysis. Association mapping detected 15 MTAs that were significantly ( p ≤ 0.0005) associated with the six stress-adaptive traits averaged across all environments and consistent in multiple individual environments. The identified MTAs were four for lamina wax, three for petiole wax, three for stem thickness, two for the flowering duration, one for the normalized difference vegetation index (NDVI), and two for the normalized pigment and chlorophyll index (NPCI). Sixteen candidate genes were identified within a 15 kb distance from either side of the markers. The detected MTAs and candidate genes have prospective use towards selecting stress-hardy pea cultivars in marker-assisted selection.

Field pea (Pisum sativum L.), a cool-season legume crop, is known for poor heat tolerance. Our previous work identified PR11-2 and PR11-90 as heat tolerant and susceptible lines in a recombinant inbred population. CDC Amarillo, a Canadian elite pea variety, was considered as another heat tolerant variety based on its similar field performance as PR11-2. This study aimed to characterize the differential transcription. Plants of these three varieties were stressed for 3 h at 38°C prior to self-pollination, and RNAs from heat stressed anthers and stipules on the same flowering node were extracted and sequenced via the Illumina NovaSeq platform for the characterization of heat responsive genes. In silico results were further validated by qPCR assay. Differentially expressed genes (DEGs) were identified at log2 |fold change (FC)| ≥ 2 between high temperature and control temperature, the three varieties shared 588 DEGs which were up-regulated and 220 genes which were down-regulated in anthers when subjected to heat treatment. In stipules, 879 DEGs (463/416 upregulation/downregulation) were consistent among varieties. The above heat-induced genes of the two plant organs were related to several biological processes i.e., response to heat, protein folding and DNA templated transcription. Ten gene ontology (GO) terms were over-represented in the consistently down-regulated DEGs of the two organs, and these terms were mainly related to cell wall macromolecule metabolism, lipid transport, lipid localization, and lipid metabolic processes. GO enrichment analysis on distinct DEGs of individual pea varieties suggested that heat affected biological processes were dynamic, and variety distinct responses provide insight into molecular mechanisms of heat-tolerance response. Several biological processes, e.g., cellular response to DNA damage stimulus in stipule, electron transport chain in anther that were only observed in heat induced PR11-2 and CDC Amarillo, and their relevance to field pea heat tolerance is worth further validation., Competing Interests: Authors RVBL and NC are employees of AgriGenome Labs Pvt. Ltd. Their commercial affiliation does not alter our adherence to PLOS ONE policies on sharing data and materials.

A validated method for B vitamin separation and quantification from lentil seeds using ultra high performance liquid chromatography-selected reaction monitoring mass spectrometry (UHPLC-SRM MS) was reported. The use of three enzymes (acid phosphatase, β-glucosidase, and rat serum) with a 4 h incubation was sufficient to convert bound B vitamins into their free forms. Twenty B vitamers were selected and a 5-min UHPLC-SRM MS method was optimized for rapid analysis. This method was applied to quantify B vitamin concentration during lentil seed germination over a 5-day period. Total B vitamins increased up to 1.5-fold on day 5 (from 39.2 µg/g to 60.6 µg/g of dry weight) comparing with dry seeds. Vitamin B5 (pantothenic acid) was the most abundant B vitamin in both dry seeds (34.2%) and in germinated seeds (17.7%-24.5% of total B vitamins); B8 (biotin) and B12 (cyanocobalamin) were not detected in lentil samples., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Field pea is a pulse that delivers high protein content, slowly digestible starch and fiber, and many vitamins and minerals, including iron. Naturally occurring plant phytic acid molecules bind iron, lowering its availability for absorption during digestion. Two low phytic acid ( lpa ) pea lines, 1-2347-144 and 1-150-81, developed by our group had 15% lower yield and 6% lower seed weight relative to their progenitor cultivar. Subsequently, we crossed the two lpa lines and two cultivars, and derived 19 promising lpa pea breeding lines; here we document their agronomic performance based on 10 replicated field trials in Saskatchewan. Seventeen of these lpa lines yielded greater than 95% of the check mean (associated cultivars) and 16 were above 98% of the check mean for 1000 seed weight. The 19 lpa lines showed 27 to 55% lower phytic acid concentration than the check mean. Iron concentrations were similar in all the lpa lines and cultivars, yet the Caco-2 human cell culture assay revealed 14 of the 19 lpa lines had 11 to 55% greater iron bioavailability than check means. Thus, a single round of plant breeding has allowed for closing the gap in performance of low phytic acid pea.

Wrinkled and round peas (two varieties each type) cultivated in two locations were milled to obtain fine and coarse wrinkled (WPF) and round pea flour (RPF). WPF exhibited markedly increased pasting viscosities at 120 and 140 °C compared with 95 °C. Overall, the pasting properties of WPF were considerably lower than those of RPF. Resistant starch (RS) contents of cooked WPF (17.2-22.2%, dsb) were significantly larger than those of RPF (7.9-11.4%), resulting from higher starch gelatinization temperatures, greater amylose contents, and presence of more protein and fiber in WPF. The two particle sizes affected the water-holding capacity (WHC) of WPF, gelatinization enthalpy changes (ΔH) of WPF and RPF, and pasting properties and starch digestibility of RPF. Pearson correlation and principal component analysis (PCA) were conducted to reveal the relationships among the techno-functional parameters of pea flours. Wrinkled pea showed promise to generate new pea flours with distinct functionality and enhanced nutritional value., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Globally, two billion people suffer from micronutrient deficiencies. Cereal grains provide more than 50% of the daily requirement of calories in human diets, but they often fail to provide adequate essential minerals and vitamins. Cereal crop production in developing countries achieved remarkable yield gains through the efforts of the Green Revolution (117% in rice, 30% in wheat, 530% in maize, and 188% in pearl millet). However, modern varieties are often deficient in essential micronutrients compared to traditional varieties and land races. Breeding for nutritional quality in staple cereals is a challenging task; however, biofortification initiatives combined with genomic tools increase the feasibility. Current biofortification breeding activities include improving rice (for zinc), wheat (for zinc), maize (for provitamin A), and pearl millet (for iron and zinc). Biofortification is a sustainable approach to enrich staple cereals with provitamin A, carotenoids, and folates. Significant genetic variation has been found for provitamin A (96-850 μg and 12-1780 μg in 100 g in wheat and maize, respectively), carotenoids (558-6730 μg in maize), and folates in rice (11-51 μg) and wheat (32.3-89.1 μg) in 100 g. This indicates the prospects for biofortification breeding. Several QTLs associated with carotenoids and folates have been identified in major cereals, and the most promising of these are presented here. Breeding for essential nutrition should be a core objective of next-generation crop breeding. This review synthesizes the available literature on folates, provitamin A, and carotenoids in rice, wheat, maize, and pearl millet, including genetic variation, trait discovery, QTL identification, gene introgressions, and the strategy of genomics-assisted biofortification for these traits. Recent evidence shows that genomics-assisted breeding for grain nutrition in rice, wheat, maize, and pearl millet crops have good potential to aid in the alleviation of micronutrient malnutrition in many developing countries., (Copyright © 2020 Ashokkumar, Govindaraj, Karthikeyan, Shobhana and Warkentin.)

The objective of this study was to examine the bioactive flavonoids and carotenoids concentration in fifteen south Indian spice and two tree nut species using high performance liquid chromatography (HPLC). Among four flavonoids, catechin concentration was the highest in all spices and nuts and ranged between 97.1 and 1745.4 µg g -1 . Quercetin concentration was the greatest in cinnamon, followed by garlic and cumin and ranged from 0.4 to 65 µg g -1 in other spices and nuts. Lutein concentration ranged from 0.1 to 102.8 µg g -1 . Of the spices and nuts studied, β-carotene concentration was highest in coriander leaves (74.7 µg g -1 ), followed by red pepper (12.5 µg g -1 ) and curry leaves (8.5 µg g -1 ). This research shows that consumption of south Indian spices and nuts could substantially benefit consumers living in regions experiencing Vitamin A and other micronutrient deficiencies.

Seed samples from 117 genetically diverse pea breeding lines were used to determine the robustness of Fourier transform mid-infrared spectroscopy (FT-MIR) for the rapid nutritional profiling of seeds. The FT-MIR results were compared to wet chemistry methods for assessing the concentrations of total protein, starch, fiber, phytic acid, and carotenoids in pea seed samples. Of the five partial least square regression models (PLSR) developed, protein, fiber and phytic acid concentrations predicted by the models exhibited correlation coefficients greater than 0.83 when compared with data obtained using the wet chemistry methods for both the calibration and validation sets. The starch PLSR model had a correlation greater than 0.75, and carotenoids had correlation of 0.71 for the validation sets. The methods implemented in this research show the novelty and usefulness of FT-MIR as a simple, fast, and cost-effective technique to determine multiple seed constituents simultaneously., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Environmental stress hampers pea productivity. To understand the genetic basis of heat resistance, a genome-wide association study (GWAS) was conducted on six stress responsive traits of physiological and agronomic importance in pea, with an objective to identify the genetic loci associated with these traits. One hundred and thirty-five genetically diverse pea accessions from major pea growing areas of the world were phenotyped in field trials across five environments, under generally ambient (control) and heat stress conditions. Statistical analysis of phenotype indicated significant effects of genotype (G), environment (E), and G × E interaction for all traits. A total of 16,877 known high-quality SNPs were used for association analysis to determine marker-trait associations (MTA). We identified 32 MTAs that were consistent in at least three environments for association with the traits of stress resistance: six for chlorophyll concentration measured by a soil plant analysis development meter; two each for photochemical reflectance index and canopy temperature; seven for reproductive stem length; six for internode length; and nine for pod number. Forty-eight candidate genes were identified within 15 kb distance of these markers. The identified markers and candidate genes have potential for marker-assisted selection towards the development of heat resistant pea cultivars.

Ethnopharmacological Relevance: Small cardamom [Elettaria cardamomum (L.) Maton. (Family: Zingiberaceae)] capsules (fruits) have been used for traditional medicine applications including for the control of asthma, teeth and gum infections, cataracts, nausea, diarrhea, as well as cardiac, digestive and kidney disorders. The versatile use of cardamom capsules has several other beneficial health effects that are relevant in light of traditional and modern pharmaceutical perspectives., Aim of the Study: This review aims to provide a critical and comprehensive evaluation of the traditional and current medical uses of E. cardamomum, and compare these applications with modern research studies. This critical review also discusses the botanical distribution, phytochemical constituents and biological activities of cardamom capsule extracts and essential oil., Materials and Methods: An online survey was conducted of the traditional uses, phytochemical composition, and pharmacological applications of cardamom essential oil (CEO) and extracts. Pertinent data were obtained from several electronic scientific databases (Science Direct, Elsevier, Web of Science, PubMed, Springer, ACS publications, Taylor and Francis, Wiley On-line Library and Google Scholar), and additional information was obtained from textbooks and local prints and scripts., Results: Cardamom fruits (capsules) are used widely as a spice and flavoring ingredient in foods, and are often recognized for their beneficial health properties. They are also used in fragrances. Phytochemical analyses have described important chemical constituents of cardamom including carbohydrates, proteins, minerals, lipids, essential oils, flavonoids, terpenoids and carotenoids. CEO has several biological roles including antioxidant, antidiabetic, antibacterial, anticancer, gastro-protective and insecticidal activities., Conclusion: The widespread availability and recommendation of synthetic compounds for addressing human health have several side effects besides higher costs. Hence, examining natural bioactive compounds is imperative. This review investigates and presents the pertinent information on cardamom and its traditional uses, as well as potential pharmacological properties of CEO and extracts. Additional research studies are needed to understand the mechanism of action of bioactive constituents., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Biofortification through plant breeding is a sustainable approach to improve the nutritional profile of food crops. The majority of the world's population depends on staple food crops; however, most are low in key micronutrients. Biofortification to improve the nutritional profile of pulse crops has increased importance in many breeding programs in the past decade. The key micronutrients targeted have been iron, zinc, selenium, iodine, carotenoids, and folates. In recent years, several biofortified pulse crops including common beans and lentils have been released by HarvestPlus with global partners in developing countries, which has helped in overcoming micronutrient deficiency in the target population. This review will focus on recent research advances and future strategies for the biofortification of pulse crops.

Genome-wide association study (GWAS) was conducted to identify loci associated with agronomic (days to flowering, days to maturity, plant height, seed yield and seed weight), seed morphology (shape and dimpling), and seed quality (protein, starch, and fiber concentrations) traits of field pea ( Pisum sativum L.). A collection of 135 pea accessions from 23 different breeding programs in Africa (Ethiopia), Asia (India), Australia, Europe (Belarus, Czech Republic, Denmark, France, Lithuania, Netherlands, Russia, Sweden, Ukraine and United Kingdom), and North America (Canada and USA), was used for the GWAS. The accessions were genotyped using genotyping-by-sequencing (GBS). After filtering for a minimum read depth of five, and minor allele frequency of 0.05, 16,877 high quality SNPs were selected to determine marker-trait associations (MTA). The LD decay (LD 1/2max,90 ) across the chromosomes varied from 20 to 80 kb. Population structure analysis grouped the accessions into nine subpopulations. The accessions were evaluated in multi-year, multi-location trials in Olomouc (Czech Republic), Fargo, North Dakota (USA), and Rosthern and Sutherland, Saskatchewan (Canada) from 2013 to 2017. Each trait was phenotyped in at least five location-years. MTAs that were consistent across multiple trials were identified. Chr5LG3_566189651 and Chr5LG3_572899434 for plant height, Chr2LG1_409403647 for lodging resistance, Chr1LG6_57305683 and Chr1LG6_366513463 for grain yield, Chr1LG6_176606388, Chr2LG1_457185, Chr3LG5_234519042 and Chr7LG7_8229439 for seed starch concentration, and Chr3LG5_194530376 for seed protein concentration were identified from different locations and years. This research identified SNP markers associated with important traits in pea that have potential for marker-assisted selection towards rapid cultivar improvement., (Copyright © 2019 Gali, Sackville, Tafesse, Lachagari, McPhee, Hybl, Mikić, Smýkal, McGee, Burstin, Domoney, Ellis, Tar'an and Warkentin.)

Core Ideas: Quantitative trait locus (QTL) analyses for carotenoids in chickpea were completed for three F 2 populations. A moderate number of QTLs and candidate genes associated with carotenoid concentration in chickpea seeds were identified. Green cotyledon color is positively associated with provitamin A carotenoids. Three F 2 populations derived from crosses between cultivars with green and yellow cotyledon colors were used to identify quantitative trait loci (QTLs) associated with carotenoid components in chickpea (Cicer arietinum L.) seeds developed by the Crop Development Centre (CDC). Carotenoids including violaxanthin, lutein, zeaxanthin, β-cryptoxanthin, and β-carotene were assessed in the F 2:3 seeds via high-performance liquid chromatography (HPLC). In the 'CDC Jade' × 'CDC Frontier' population, 1068 bin markers derived from the 50K Axiom CicerSNP array were mapped onto eight linkage groups (LGs). Eight QTLs, including two each for β-carotene and zeaxanthin and one each for total carotenoids, β-cryptoxanthin, β-carotene, and violaxanthin were identified in this population. In the 'CDC Cory' × 'CDC Jade' population, 694 bin markers were mapped onto eight LGs and one partial LG. Quantitative trait loci for β-cryptoxanthin, β-carotene, violaxanthin, lutein, and total carotenoids were identified on LG8. A map with eight LGs was developed from 581 bin markers in the third population derived from the 'ICC4475' × 'CDC Jade' cross. One QTL for β-carotene and four QTLs, one each for β-cryptoxanthin, β-carotene, lutein, and total carotenoids, were identified in this population. The highest phenotypic variation explained by the QTLs was for β-carotene, which ranged from 58 to 70% in all three populations. A major gene for cotyledon color was mapped on LG8 in each population. A significant positive correlation between cotyledon color and carotenoid concentration was observed. Potential candidate genes associated with carotenoid components were obtained and their locations on the chickpea genome are presented., (© 2019 The Author(s).)

We report the first annotated chromosome-level reference genome assembly for pea, Gregor Mendel's original genetic model. Phylogenetics and paleogenomics show genomic rearrangements across legumes and suggest a major role for repetitive elements in pea genome evolution. Compared to other sequenced Leguminosae genomes, the pea genome shows intense gene dynamics, most likely associated with genome size expansion when the Fabeae diverged from its sister tribes. During Pisum evolution, translocation and transposition differentially occurred across lineages. This reference sequence will accelerate our understanding of the molecular basis of agronomically important traits and support crop improvement.

Folates are important micronutrients in lentils (Lens culinaris Medik.). In this work, the folate extraction workflow in ascorbate-containing buffer was optimized and validated, and the concentrations of eight folate monoglutamates in cultivated and six wild lentil species, grown under field or greenhouse conditions, were quantified by ultra-performance liquid chromatography and mass spectrometry (UPLC-MS). In general wild lentil species had higher folate concentrations than cultivated genotypes. Lens tomentosus had the highest folate concentration with median values of 439.7 and 360.9 μg/100 g in the field and greenhouse, followed by Lens orientalis with 416.6 and 327.6 μg/100 g, respectively. A significant effect (P < 0.05) of growing conditions was observed in four out of six wild lentil species, with seeds from the field having higher folate concentration (6% to 45%) compared with the greenhouse. MeFox, an oxidation product of 5-methyltetrahydrofolate, was present in all lentil species at concentrations 2.2 to 5.6 times higher than the total folates., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Field pea (Pisum sativum), a major grain legume crop, is autogamous and adapted to temperate climates. The objectives of this study were to investigate effects of high temperature stress on stamen chemical composition, anther dehiscence, pollen viability, pollen interactions with pistil and ovules, and ovule growth and viability. Two cultivars ("CDC Golden" and "CDC Sage") were exposed to 24/18°C (day/night) continually or to 35/18°C for 4 or 7 days. Heat stress altered stamen chemical composition, with lipid composition of "CDC Sage" being more stable compared with "CDC Golden." Heat stress reduced pollen viability and the proportion of ovules that received a pollen tube. After 4 days at 35°C, pollen viability in flower buds decreased in "CDC Golden," but not in "CDC Sage." After 7 days, partial to full failure of anthers to dehisce resulted in subnormal pollen loads on stigmas. Although growth (ovule size) of fertilized ovules was stimulated by 35°C, heat stress tended to decrease ovule viability. Pollen appears susceptible to stress, but not many grains are needed for successful fertilization. Ovule fertilization and embryos are less susceptible to heat, but further research is warranted to link the exact degree of resilience to stress intensity., (© 2018 John Wiley & Sons Ltd.)

Background: The objective of this research was to map quantitative trait loci (QTLs) of multiple traits of breeding importance in pea (Pisum sativum L.). Three recombinant inbred line (RIL) populations, PR-02 (Orb x CDC Striker), PR-07 (Carerra x CDC Striker) and PR-15 (1-2347-144 x CDC Meadow) were phenotyped for agronomic and seed quality traits under field conditions over multiple environments in Saskatchewan, Canada. The mapping populations were genotyped using genotyping-by-sequencing (GBS) method for simultaneous single nucleotide polymorphism (SNP) discovery and construction of high-density linkage maps., Results: After filtering for read depth, segregation distortion, and missing values, 2234, 3389 and 3541 single nucleotide polymorphism (SNP) markers identified by GBS in PR-02, PR-07 and PR-15, respectively, were used for construction of genetic linkage maps. Genetic linkage groups were assigned by anchoring to SNP markers previously positioned on these linkage maps. PR-02, PR-07 and PR-15 genetic maps represented 527, 675 and 609 non-redundant loci, and cover map distances of 951.9, 1008.8 and 914.2 cM, respectively. Based on phenotyping of the three mapping populations in multiple environments, 375 QTLs were identified for important traits including days to flowering, days to maturity, lodging resistance, Mycosphaerella blight resistance, seed weight, grain yield, acid and neutral detergent fiber concentration, seed starch concentration, seed shape, seed dimpling, and concentration of seed iron, selenium and zinc. Of all the QTLs identified, the most significant in terms of explained percentage of maximum phenotypic variance (PV max ) and occurrence in multiple environments were the QTLs for days to flowering (PV max  = 47.9%), plant height (PV max  = 65.1%), lodging resistance (PV max  = 35.3%), grain yield (PV max  = 54.2%), seed iron concentration (PV max  = 27.4%), and seed zinc concentration (PV max  = 43.2%)., Conclusion: We have identified highly significant and reproducible QTLs for several agronomic and seed quality traits of breeding importance in pea. The QTLs identified will be the basis for fine mapping candidate genes, while some of the markers linked to the highly significant QTLs are useful for immediate breeding applications.

Ascochyta blight (AB) is an important disease of pea which can cause severe grain yield loss under wet conditions. In our previous study, we identified two quantitative trait loci (QTLs) abIII-1 and abI-IV-2 for AB resistance and these QTLs were consistent across locations and/or years in an inter-specific pea population (PR-19) developed from a cross between Alfetta ( Pisum sativum ) and P651 ( P . fulvum ). The objectives of this study were to fine map the abIII-1 and abI-IV-2 QTLs using a high density single nucleotide polymorphism (SNP)-based genetic linkage map and analyze identified markers in heterogeneous inbred family (HIF) populations. Selective genotyping of 51 PR-19 recombinant inbred lines was performed using genotyping-by-sequencing (GBS) and the resulting high density genetic linkage map was used to identify eight new SNP markers within the abI-IV-2 QTL, whereas no additional SNPs were identified within the abIII-1 QTL. Two HIF populations HIF-224 (143 lines) and HIF-173 (126 lines) were developed from F 6 RILs PR-19-224 and PR-19-173, respectively. The HIF populations evaluated under field conditions in 2015 and 2016 showed a wide range of variation for reaction to AB resistance. Lodging score had significant positive ( P < 0.001) correlation with AB scores. HIFs were genotyped using SNP markers within targeted QTLs. The genotypic and phenotypic data of the HIFs were used to identify two new QTLs, abI-IV-2.1 and abI-IV-2.2 for AB resistance within the abI-IV-2 QTL. These QTLs individually explained 5.5 to 14% of the total phenotypic variation. Resistance to lodging was also associated with these two QTLs. Identified SNP markers will be useful in marker assisted selection for development of pea cultivars with improved AB resistance.

The objective of this study was to determine the molecular structure and properties of recently released cultivars of field peas [CDC Golden (CDCG), Abarth (ABAR), CDC Patrick (CDCP) and CDC Amarillo (CDCA)] grown at different locations in Saskatchewan, Canada. Starch yield (on whole seed basis), apparent amylose, total lipid and specific surface area were in the range 34-37%, 38.2-42.6%, 1.07-1.38% and 0.31-0.38m 2 /g, respectively. The proportion of short (DP 6-12) amylopectin chains, amylopectin branching density, molecular order, crystallinity, crystalline heterogeneity, gelatinization transition temperatures, pasting temperatures, peak viscosity, extent of acid hydrolysis, and resistant starch content were higher in CDCG and ABAR. However, amylopectin long chains (DP 13-26), average chain length and thermal stability were higher in CDCP and CDCA. The results of this study showed that differences in physicochemical properties among cultivars were mainly influenced by amylopectin chain length distribution, amylopectin branching density and co-crystallization of amylose with amylopectin., (Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.)

In western Canada, chickpea (Cicer arietinum L.) production is challenged by short growing seasons and infestations with ascochyta blight. Research was conducted to determine the genetic basis of the association between flowering time and reaction to ascochyta blight in chickpea. Ninety-two chickpea recombinant inbred lines (RILs) developed from a cross between ICCV 96029 and CDC Frontier were evaluated for flowering responses and ascochyta blight reactions in growth chambers and fields at multiple locations and during several years. A wide range of variation was exhibited by the RILs for days to flower, days to maturity, node of first flowering, plant height, and ascochyta blight resistance. Moderate to high broad sense heritability was estimated for ascochyta blight reaction (H(2) = 0.14-0.34) and for days to flowering (H(2) = 0.45-0.87) depending on the environments. Negative correlations were observed among the RILs for days to flowering and ascochyta blight resistance, ranging from r = -0.21 (P < 0.05) to -0.58 (P < 0.0001). A genetic linkage map consisting of eight linkage groups was developed using 349 SNP markers. Seven QTLs for days to flowering were identified that individually explained 9%-44% of the phenotypic variation. Eight QTLs were identified for ascochyta blight resistance that explained phenotypic variation ranging from 10% to 19%. Clusters of QTLs for days to flowering and ascochyta blight resistances were found on chromosome 3 at the interval of 8.6-23.11 cM and on chromosome 8 at the interval of 53.88-62.33 cM.

Photoperiod is one of the major environmental factors determining time to flower initiation and first flower appearance in plants. In chickpea, photoperiod sensitivity, expressed as delayed to flower under short days (SD) as compared to long days (LD), may change with the growth stage of the crop. Photoperiod-sensitive and -insensitive phases were identified by experiments in which individual plants were reciprocally transferred in a time series from LD to SD and vice versa in growth chambers. Eight chickpea accessions with differing degrees of photoperiod sensitivity were grown in two separate chambers, one of which was adjusted to LD (16 h light/8 h dark) and the other adjusted to SD (10 h light/14 h dark), with temperatures of 22/16°C (12 h light/12 h dark) in both chambers. The accessions included day-neutral (ICCV 96029 and FLIP 98-142C), intermediate (ICC 15294, ICC 8621, ILC 1687, and ICC 8855), and photoperiod-sensitive (CDC Corinne and CDC Frontier) responses. Control plants were grown continuously under the respective photoperiods. Reciprocal transfers of plants between the SD and LD photoperiod treatments were made at seven time points after sowing, customized for each accession based on previous data. Photoperiod sensitivity was detected in intermediate and photoperiod-sensitive accessions. For the day-neutral accession, ICCV 96029, there was no significant difference in the number of days to flowering of the plants grown under SD and LD as well as subsequent transfers. In photoperiod-sensitive accessions, three different phenological phases were identified: a photoperiod-insensitive pre-inductive phase, a photoperiod-sensitive inductive phase, and a photoperiod-insensitive post-inductive phase. The photoperiod-sensitive phase extends after flower initiation to full flower development. Results from this research will help to develop cultivars with shorter pre-inductive photoperiod-insensitive and photoperiod-sensitive phases to fit to regions with short growing seasons.

Single nucleotide polymorphism (SNP) arrays represent important genotyping tools for innovative strategies in both basic research and applied breeding. Pea is an important food, feed and sustainable crop with a large (about 4.45 Gbp) but not yet available genome sequence. In the present study, 12 pea recombinant inbred line populations were genotyped using the newly developed GenoPea 13.2K SNP Array. Individual and consensus genetic maps were built providing insights into the structure and organization of the pea genome. Largely collinear genetic maps of 3918-8503 SNPs were obtained from all mapping populations, and only two of these exhibited putative chromosomal rearrangement signatures. Similar distortion patterns in different populations were noted. A total of 12 802 transcript-derived SNP markers placed on a 15 079-marker high-density, high-resolution consensus map allowed the identification of ohnologue-rich regions within the pea genome and the localization of local duplicates. Dense syntenic networks with sequenced legume genomes were further established, paving the way for the identification of the molecular bases of important agronomic traits segregating in the mapping populations. The information gained on the structure and organization of the genome from this research will undoubtedly contribute to the understanding of the evolution of the pea genome and to its assembly. The GenoPea 13.2K SNP Array and individual and consensus genetic maps are valuable genomic tools for plant scientists to strengthen pea as a model for genetics and physiology and enhance breeding., (© 2015 The Authors The Plant Journal © 2015 John Wiley & Sons Ltd.)

Pea (Pisum sativum L.) is an annual cool-season legume and one of the oldest domesticated crops. Dry pea seeds contain 22-25% protein, complex starch and fiber constituents, and a rich array of vitamins, minerals, and phytochemicals which make them a valuable source for human consumption and livestock feed. Dry pea ranks third to common bean and chickpea as the most widely grown pulse in the world with more than 11 million tons produced in 2013. Pea breeding has achieved great success since the time of Mendel's experiments in the mid-1800s. However, several traits still require significant improvement for better yield stability in a larger growing area. Key breeding objectives in pea include improving biotic and abiotic stress resistance and enhancing yield components and seed quality. Taking advantage of the diversity present in the pea genepool, many mapping populations have been constructed in the last decades and efforts have been deployed to identify loci involved in the control of target traits and further introgress them into elite breeding materials. Pea now benefits from next-generation sequencing and high-throughput genotyping technologies that are paving the way for genome-wide association studies and genomic selection approaches. This review covers the significant development and deployment of genomic tools for pea breeding in recent years. Future prospects are discussed especially in light of current progress toward deciphering the pea genome.

Low phytic acid (lpa) crops are low in phytic acid and high in inorganic phosphorus (Pi). In this study, two lpa pea genotypes, 1-150-81, 1-2347-144, and their progenitor CDC Bronco were grown in field trials for two years. The lpa genotypes were lower in IP₆ and higher in Pi when compared to CDC Bronco. The total P concentration was similar in lpa genotypes and CDC Bronco throughout the seed development. The action of myo-inositol phosphate synthase (MIPS) (EC 5.5.1.4) is the first and rate-limiting step in the phytic acid biosynthesis pathway. Aiming at understanding the genetic basis of the lpa mutation in the pea, a 1530 bp open reading frame of MIPS was amplified from CDC Bronco and the lpa genotypes. Sequencing results showed no difference in coding sequence in MIPS between CDC Bronco and lpa genotypes. Transcription levels of MIPS were relatively lower at 49 days after flowering (DAF) than at 14 DAF for CDC Bronco and lpa lines. This study elucidated the rate and accumulation of phosphorus compounds in lpa genotypes. The data also demonstrated that mutation in MIPS was not responsible for the lpa trait in these pea lines.

Key Message: Gene-based SNPs were identified and mapped in pea using five recombinant inbred line populations segregating for traits of agronomic importance. Pea (Pisum sativum L.) is one of the world's oldest domesticated crops and has been a model system in plant biology and genetics since the work of Gregor Mendel. Pea is the second most widely grown pulse crop in the world following common bean. The importance of pea as a food crop is growing due to its combination of moderate protein concentration, slowly digestible starch, high dietary fiber concentration, and its richness in micronutrients; however, pea has lagged behind other major crops in harnessing recent advances in molecular biology, genomics and bioinformatics, partly due to its large genome size with a large proportion of repetitive sequence, and to the relatively limited investment in research in this crop globally. The objective of this research was the development of a genome-wide transcriptome-based pea single-nucleotide polymorphism (SNP) marker platform using next-generation sequencing technology. A total of 1,536 polymorphic SNP loci selected from over 20,000 non-redundant SNPs identified using deep transcriptome sequencing of eight diverse Pisum accessions were used for genotyping in five RIL populations using an Illumina GoldenGate assay. The first high-density pea SNP map defining all seven linkage groups was generated by integrating with previously published anchor markers. Syntenic relationships of this map with the model legume Medicago truncatula and lentil (Lens culinaris Medik.) maps were established. The genic SNP map establishes a foundation for future molecular breeding efforts by enabling both the identification and tracking of introgression of genomic regions harbouring QTLs related to agronomic and seed quality traits.

Background: In the whole genome sequencing, genetic map provides an essential framework for accurate and efficient genome assembly and validation. The main objectives of this study were to develop a high-density genetic map using RAD-Seq (Restriction-site Associated DNA Sequencing) genotyping-by-sequencing (RAD-Seq GBS) and Illumina GoldenGate assays, and to examine the alignment of the current map with the kabuli chickpea genome assembly., Results: Genic single nucleotide polymorphisms (SNPs) totaling 51,632 SNPs were identified by 454 transcriptome sequencing of Cicer arietinum and Cicer reticulatum genotypes. Subsequently, an Illumina GoldenGate assay for 1,536 SNPs was developed. A total of 1,519 SNPs were successfully assayed across 92 recombinant inbred lines (RILs), of which 761 SNPs were polymorphic between the two parents. In addition, the next generation sequencing (NGS)-based GBS was applied to the same population generating 29,464 high quality SNPs. These SNPs were clustered into 626 recombination bins based on common segregation patterns. Data from the two approaches were used for the construction of a genetic map using a population derived from an intraspecific cross. The map consisted of 1,336 SNPs including 604 RAD recombination bins and 732 SNPs from Illumina GoldenGate assay. The map covered 653 cM of the chickpea genome with an average distance between adjacent markers of 0.5 cM. To date, this is the most extensive genetic map of chickpea using an intraspecific population. The alignment of the map with the CDC Frontier genome assembly revealed an overall conserved marker order; however, a few local inconsistencies within the Cicer arietinum pseudochromosome 1 (Ca1), Ca5 and Ca8 were detected. The map enabled the alignment of 215 unplaced scaffolds from the CDC Frontier draft genome assembly. The alignment also revealed varying degrees of recombination rates and hotspots across the chickpea genome., Conclusions: A high-density genetic map using RAD-Seq GBS and Illumina GoldenGate assay was developed and aligned with the existing kabuli chickpea draft genome sequence. The analysis revealed an overall conserved marker order, although some localized inversions between draft genome assembly and the genetic map were detected. The current analysis provides an insight of the recombination rates and hotspots across the chickpea genome.

Chickpea (Cicer arietinum L.) is the world's second most important pulse crop after common bean. Chickpea has historically been an important daily staple in the diet of millions of people, especially in the developing countries. Current chickpea breeding programs have mainly been directed toward high yield, biotic and abiotic stress resilience that has increased global production, but less attention has been directed toward improving micronutrient concentrations in seeds. In an effort to develop micronutrient-dense chickpea lines, a study to examine the variability and to identify SNP alleles associated with seed iron and zinc concentrations was conducted using 94 diverse accessions of chickpea. The results indicated that there is substantial variability present in chickpea germplasm for seed iron and zinc concentrations. In the current set of germplasm, zinc is negatively correlated with grain yield across all locations and years; whereas the negative correlation between iron and grain yield was only significant at the Elrose locality. Eight SNP loci associated with iron and (or) zinc concentrations in chickpea seeds were identified. One SNP located on chromosome 1 (chr1) is associated with both iron and zinc concentrations. On chr4, three SNPs associated with zinc concentration and two SNPs for iron concentration were identified. Two additional SNP loci, one on chr6 and the other on chr7, were also found to be associated with iron and zinc concentrations, respectively. The results show potential opportunity for molecular breeding for improvement of seed iron and zinc concentrations in chickpea.

Ascochyta blight (AB) caused by the fungus Ascochyta rabiei Pass. Lab. is one of the major diseases of chickpea worldwide and a constraint to production in western Canada. The use of varieties with high levels of resistance is considered the most economical solution for long-term ascochyta blight management in chickpea. QTL for resistance to ascochyta blight have been identified in chickpea. The availability of molecular markers associated with QTL for ascochyta blight resistant and double podding provides an opportunity to apply marker-assisted backcrossing to introgress the traits into adapted chickpea cultivars. In the present study, molecular markers that were linked to the QTL for ascochyta blight resistance and the double podding trait, and those unlinked to the resistance were used in foreground and background selection, respectively, in backcrosses between moderately resistant donors (CDC Frontier and CDC 425-14) and the adapted varieties (CDC Xena, CDC Leader and FLIP98-135C). The strategy included two backcrosses and selection for two QTL for ascochyta blight resistance and a locus associated with double podding. The fixation of the elite genetic background was monitored with 16-22 SSR markers to accelerate restoration of the genetic background at each backcross. By the BC2F1 generation, plants with improved ascochyta blight resistance and double podding were identified. The selected plants possessed the majority of elite parental type SSR alleles on all fragments analyzed except the segment of LG 4, LG 6 and LG 8 that possessed the target QTL. The results showed that the adapted variety could be efficiently converted into a variety with improved resistance in two backcross generations.

Background: Consumption of pulse crops, including field pea, is considered effective for a healthy diet. Hulls (seed coats) play an important role for protection of the cotyledon and embryo, but also as mediating positive effects on health outcomes. The biochemical attributes of field pea hulls were thus assessed to determine the occurrence of specific phytochemicals and their genotypic variability., Results: Sequestered bioproducts in mature hulls predominantly consisted of trans-lutein and chlorophylls a and b. Trace amounts of other carotenoid and pheophytin metabolites were identified. In developing hulls, violaxanthin, neoxanthin, lutein, zeaxanthin, chlorophylls a and b and β-carotene were detected. Genotypic differences in the accumulation of lutein and chlorophylls a and b were observed over years and locations. Polyphenolics and hydroxybenzoic acids were detected in the 'dun' and 'maple' field pea types-the only genotypes to have pigmented hulls. Unextractable patches of condensed tannin influenced the visual uniformity of the maple and dun genotypes, CDC Rocket and CDC Dundurn., Conclusions: Within the yellow and green market classes, carotenoid and chlorophyll accumulation was consistent. Green cotyledon varieties sequestered higher concentrations of lutein than the yellow cotyledon varieties. Maple and dun types were more variable, reflective of different selection criteria. The occurrence of flavonoid-related compounds was correlated only with pigmented seed coat genotypes. The dietary potential of the chlorophylls and carotenoids that accumulated in the hulls split from the green and yellow field pea types is discussed as a value-added prospect in food supplements., (Copyright © 2012 Society of Chemical Industry.)

Background: The effects of genotype and environment and their interaction on the concentrations of starch and protein in, and the amylose content and thermal and pasting properties of starch from, pea and fababean are not well known., Results: Differences due to genotype were observed in the concentrations of starch and protein in pea and fababean, in the onset temperature (To) and peak temperature (Tp) of gelatinization of fababean starch, and in the pasting, trough, cooling and final viscosities of pea starch and fababean starch. Significant two-way interactions (location × genotype) were observed for the concentration of starch in fababean and the amylose content, To, endothermic enthalpy of gelatinization (ΔH) and trough viscosity of fababean starch. Significant three-way interactions (location × year × genotype) were observed for the concentration of starch in pea and the pasting, trough, cooling and final viscosities of pea starch., Conclusion: Differences observed in the concentrations of starch and protein in pea and fababean were sufficient to be of practical significance to end-users, but the relatively small differences in amylose content and physicochemical properties of starch from pea and fababean were not., (Copyright © 2011 Society of Chemical Industry.)

Ascochyta blight (AB) caused by Ascochyta rabiei (teleomorph, Didymella rabiei) Pass. Lab. is an important fungal disease of chickpea worldwide. Only moderate sources of resistance are available within the cultivated species and we hypothesized that the available sources may carry different genes for resistance, which could be pyramided to improve field resistance to AB. Four divergent moderately resistant cultivars CDC Frontier, CDC Luna, CDC Corinne, and Amit were each crossed to a highly susceptible germplasm ICCV 96029. Parents, F(1) and F(2) generations were evaluated under controlled conditions for their reactions to AB. A total of 144 simple sequence repeat (SSR) markers were first mapped to eight linkage groups (LG) for the CDC Frontier x ICCV 96029 population. Then based on the evidence from this population, 76, 61, and 42 SSR markers were systematically chosen and mapped in CDC Luna, CDC Corinne, and Amit populations, respectively. Frequency distributions of the AB rating in the F(2) generation varied among the four populations. Composite interval mapping revealed five QTLs (QTL1-5), one on each of LG 2, 3, 4, 6, and 8, respectively, distributed across different sources, controlling resistance to AB. CDC Frontier contained QTL2, 3, and 4 that simultaneously accounted for 56% of phenotypic variations. CDC Luna contained QTL 1 and 3. CDC Corinne contained QTL 3 and 5, while only QTL 2 was identified in Amit. Altogether these QTL explained 48, 38, and 14% of the estimated phenotypic variations in CDC Luna, CDC Corinne, and Amit populations, respectively. The results suggested that these QTLs could be combined into a single genotype to enhance field resistance to AB.

This is the first report on the production of double-haploid chickpea embryos and regenerated plants through anther culture using Canadian cultivar CDC Xena (kabuli) and Australian cultivar Sonali (desi). Maximum anther induction rates were 69% for Sonali and 63% for CDC Xena. Under optimal conditions, embryo formation occurred within 15-20 days of culture initiation with 2.3 embryos produced per anther for CDC Xena and 2.0 embryos per anther for Sonali. For anther induction, the following stress treatments were used: (1) flower clusters were treated at 4 degrees C for 4 days, (2) anthers were subjected to electric shock treatment of three exponentially decaying pulses of 50-400 V with 25 microF capacitance and 25 Omega resistance, (3) anthers were centrifuged at 168-1,509g for 2-15 min, and finally (4) anthers were cultured for 4 days in high-osmotic pressure (563 mmol) liquid medium. Anthers were then transferred to a solid embryo development medium and, 15-20 days later, embryo development was observed concomitant with a small amount of callus growth of 0.1-3 mm. Anther-derived embryos were regenerated on plant regeneration medium. Electroporation treatment of anthers enhanced root formation, which is often a major hurdle in legume regeneration protocols. Cytological studies using DAPI staining showed a wide range of ploidy levels from haploid to tetraploid in 10-30-day-old calli. Flow cytometric analysis of calli, embryos and regenerated plants showed haploid profiles and/or spontaneous doubling of the chromosomes during early regeneration stages.

In vitro starch digestibility, expected glycemic index (eGI), and thermal and pasting properties of flours from pea, lentil and chickpea grown in Canada under identical environmental conditions were investigated. The protein content and gelatinization transition temperatures of lentil flour were higher than those of pea and chickpea flours. Chickpea flour showed a lower amylose content (10.8-13.5%) but higher free lipid content (6.5-7.1%) and amylose-lipid complex melting enthalpy (0.7-0.8J/g). Significant differences among cultivars within the same species were observed with respect to swelling power, gelatinization properties, pasting properties and in vitro starch digestibility, especially chickpea flour from desi (Myles) and kabuli type (FLIP 97-101C and 97-Indian2-11). Lentil flour was hydrolyzed more slowly and to a lesser extent than pea and chickpea flours. The amount of slowly digestible starch (SDS) in chickpea flour was the highest among the pulse flours, but the resistant starch (RS) content was the lowest. The eGI of lentil flour was the lowest among the pulse flours., (Crown Copyright © 2008. Published by Elsevier Ltd. All rights reserved.)

Ascochyta blight, caused by the fungus Ascochyta rabiei (Pass.) Lab., is one of the most devastating diseases of chickpea (Cicer arietinum L.) worldwide. Research was conducted to map genetic factors for resistance to ascochyta blight using a linkage map constructed with 144 simple sequence repeat markers and 1 morphological marker (fc, flower colour). Stem cutting was used to vegetatively propagate 186 F2 plants derived from a cross between Cicer arietinum L. 'ICCV96029' and 'CDC Frontier'. A total of 556 cutting-derived plants were evaluated for their reaction to ascochyta blight under controlled conditions. Disease reaction of the F1 and F2 plants demonstrated that the resistance was dominantly inherited. A Fain's test based on the means and variances of the ascochyta blight reaction of the F3 families showed that a few genes were segregating in the population. Composite interval mapping identified 3 genomic regions that were associated with the reaction to ascochyta blight. One quantitative trait locus (QTL) on each of LG3, LG4, and LG6 accounted for 13%, 29%, and 12%, respectively, of the total estimated phenotypic variation for the reaction to ascochyta blight. Together, these loci controlled 56% of the total estimated phenotypic variation. The QTL on LG4 and LG6 were in common with the previously reported QTL for ascochyta blight resistance, whereas the QTL on LG3 was unique to the current population.

A study was conducted to evaluate the contribution of location, N application and Rhizobium seed inoculation to variations in seed protein content and amino acid (AA) composition of field peas. The magnitude of AA variations with protein level and the nature of the relationships that are involved were determined. Regression equations to predict AAs from protein were developed for the cultivar Bohatyr. The experiments were carried out at two locations in southern Manitoba in 1994. The levels of N fertilization investigated were: 56, 75, 100, 125, 150, 200, 250 and 300 kg/ha. At each level of N application, seeds planted were either Rhizobium inoculated or not inoculated. The combination of location, fertilizer treatments and inoculation yielded 192 samples for chemical analyses. The samples were analyzed for dry matter (DM), N and AA contents. Location and N fertilization had significant (p < or = 0.001) effects on seed protein content and AA composition. Seed protein content increased with increasing levels of N application. The response of protein to fertilization was not the same in both locations as evidenced from the presence of interaction (p < or = 0.01) between location and N application. Except for methionine and cystine, percent AAs in DM increased with increasing levels of N application. The effects of N application on the concentrations of methionine and cystine were not consistent. On protein basis, the concentrations of AAs decreased with increasing levels of N application. The only exception was arginine which strongly increased in concentration. There was no effect (p > or = 0.05) of seed inoculation observed in this study. Strong positive correlations (r > 0.80) between seed protein content and AA concentrations expressed as percent of DM were found for all AAs except for methionine (r = 0.76) and cystine (r = 0.51). When AA concentrations are expressed as g per 16 g N, 15 of the 17 AAs were negatively correlated to seed protein content. Only arginine (r = 0.78) and aspartic acid (r = 0.17) had positive correlations. The regression equations developed from this study could be used to predict the concentrations of AAs except methionine and cystine for the cultivar Bohatyr once the protein content is known.

Two different types of T-DNA insert were found in tobacco plants transformed with Agrobacterium tumefaciens. High-expressing (H) types had one copy of the T-DNA at a locus and produced high expression of the transgene uidA, as measured by uidA RNA levels and beta-glucuronidase activity; low-expressing (L) types had inverted repeats of the T-DNA at a locus and produced low uidA expression. H-types from different transformants acted additively, and cross-fertilization between two different homozygous transformants with H-type inserts produced F1 plants with GUS activity that equalled the parents and individual F2 plants with 50%, 100%, 150% and 200% of parental values. However, the L-type inserts worked in trans to suppress uidA expression from H-type inserts when both were present in the same genome. Hence when a transformant homozygous for the L-type insert was crossed to one homozygous for the H-type, all plants in the F1 and F2 generations with both types of insert had low GUS activity while F2 segregants that only had the H-type inserts had high GUS activity again. Suppression of the H-type gene was associated with increased methylation of the insert. Particle acceleration was used to introduce further copies of uidA into tissues of the transformants. Regardless of the promoter used, those plants with endogenous L-type inserts showed none of the distinct loci of GUS activity readily visible in material with no inserts, showing that L-type inserts could suppress not only the uidA expression of genomic homologues, but also of copies added in vitro.