Bianyun Yu

Rapid quantitative characterization of protein-rich agricultural materials remains challenging due to the molecular and elemental complexity of plant-derived matrices. In this study, external-cavity quantum cascade laser-based mid-infrared (QCL-MIR) spectroscopy and laser-induced breakdown spectroscopy (LIBS) were investigated as complementary approaches for predicting protein and elemental composition in pea flour (Pisum sativum L.) samples. QCL-MIR spectroscopy primarily probed molecular vibrational information, whereas LIBS directly monitored elemental composition through plasma-generated atomic and ionic emission signatures. To address the high dimensionality and nonlinear relationships present in both spectroscopic datasets, advanced chemometric strategies combining Uninformative Variable Elimination (UVE), Partial Least Squares (PLS) regression, and Extreme Learning Machine (ELM) modeling were implemented. Given the limited dataset size (N = 20), predictive performance was assessed using a concentration-stratified five-fold prediction strategy. The optimized UVE–PLS–ELM approach improved predictive performance compared with conventional linear PLS models for both techniques. QCL-MIR spectroscopy showed strong predictive performance within the present dataset for total protein content (R² = 0.97) and also provided promising predictions of phosphorus and potassium through indirect correlations with sample composition. LIBS also demonstrated strong calibration and predictive capability for total protein content (R² = 0.93) and good predictive performance for Na, Mg, Ca, Mn, and Fe. However, LIBS performance decreased at concentration extremes, particularly for high-concentration elements (K and P) and trace elements such as Zn and Cu. These results demonstrate the complementary strengths of QCL-MIR and LIBS and highlight the potential of nonlinear chemometric approaches for accurate compositional analysis of protein-rich agricultural materials within the present limited dataset.

Dormancy release and germination of the seed are two separate, but continuous phases controlled by both external (e.g., light and temperature) and internal (e.g., circadian clock and hormones) cues. In eudicot seeds, the endosperm tissues play a key role in dormancy release and germination through dynamic modulation of wall components and biomechanics. However, the mode of action by which the circadian oscillator influences dormancy release by modulation of endosperm wall biomechanics remains elusive. SCARECROW‐LIKE15 (SCL15) represses embryonic gene expression in seedlings through interaction with HISTONE DEACETYLASE19 (HDA19) in Arabidopsis thaliana . Here, we report that SCL15 plays a positive role in primary dormancy release, which is associated with gene expression changes in circadian, abscisic acid, auxin and cell wall (CW) remodelling pathways, based on studies using SCL15 mutant and Napin promoter‐driven SCL15 expression lines. SCL15 was found to affect the expression of genes whose products modify endosperm wall biomechanical features, possibly through regulation of local auxin accumulation and evening‐phased clock components. RNA‐seq analysis supported the notion that dormancy release is associated with changes in the expression of genes associated with circadian and hormone‐mediated pathways, which in turn affect CW structure.

Endosperm development is crucial for embryo growth and seed maturation. LEAFY COTYLEDON1 (LEC1), expressed in both endosperm and embryo, serves as a key regulator of seed development, orchestrating processes such as embryogenesis and seed maturation. LEC1 expression in the endosperm is detectable within a day after fertilization, yet its specific regulatory networks and developmental functions in this tissue remain unclear. To address this, we employed a modified INTACT system to isolate endosperm nuclei and performed ChIP-seq to map the genome-wide binding profile of LEC1 in developing endosperm. Integrating ChIP-seq with transcriptomic analyses, we uncover a critical role for LEC1 in regulating diverse biological pathways. Differential gene expression analysis in the endosperms of lec1 mutant and wild type shows substantial changes, particularly in genes involved in secondary cell wall biogenesis, photosynthesis, and lipid metabolism. Notably, LEC1’s regulatory networks in the endosperm shift significantly after cellularization, with distinct genes being activated in the cellular and degeneration stages. The absence of LEC1 causes significant alterations in endosperm metabolism, particularly affecting storage lipid fatty acid composition. These findings provide insights into the essential role of LEC1 in endosperm development and its broader impact on seed formation.

In soilborne diseases, the plant–pathogen interaction begins as soon as the seed germinates and develops into a seedling. Aphanomyces euteiches, an oomycete, stays dormant in soil and is activated by sensing the host through chemical signals present in the root exudates. The composition of plant exudates may, thus, play an important role during the early phase of infection. To better understand the role of root exudates in plant resistance, we investigated the interaction between partially resistant lines (PI660736 and PI557500) and susceptible pea cultivars (CDC Meadow and AAC Chrome) against A. euteiches during the pre-invasion phase. The root exudates of the two sets of cultivars clearly differed from each other in inducing oospore germination. PI557500 root exudate not only had diminished induction but also inhibited the oospore germination. The contrast between the root exudates of resistant and susceptible cultivars was reflected in their metabolic profiles. Data from fractionation and oospore germination inhibitory experiments identified a group of saponins that accumulated differentially in susceptible and resistant cultivars. We detected 56 saponins and quantified 44 of them in pea root and 30 from root exudate; the majority of them, especially soyasaponin I and dehydrosoyasaponin I with potent in vitro inhibitory activities, were present in significantly higher amounts in both roots and root exudates of PI660736 and PI557500 compared with Meadow and Chrome. Our results provide evidence for saponins as deterrents against A. euteiches, which might have contributed to the resistance against root rot in the studied pea cultivars. [ ] Copyright © 2024 His Majesty the King in Right of Canada, as represented by the Minister of Agriculture and Agri-Food Canada and the National Research Council of Canada. This is an open access article distributed under the CC BY 4.0 International license .

Pulses are a group of leguminous crops that are harvested solely for their dry seeds. As the demand for plant-based proteins grows, pulses are becoming important food crops worldwide. In addition to being a rich source of nutrients, pulses also contain saponins that are traditionally considered anti-nutrients, and impart bitterness and astringency. Saponins are plant secondary metabolites with great structural and functional diversity. Given their diverse functional properties and biological activities, both undesirable and beneficial, saponins have received growing attention. It can be expected that redirecting metabolic fluxes to control the saponin levels and produce desired saponins would be an effective approach to improve the nutritional and sensory quality of the pulses. However, little effort has been made toward understanding saponin biosynthesis in pulses, and, thus there exist sizable knowledge gaps regarding its pathway and regulatory network. In this paper, we summarize the research progress made on saponin biosynthesis in pulses. Additionally, phylogenetic relationships of putative biosynthetic enzymes among multiple pulse species provide a glimpse of the evolutionary routes and functional diversification of saponin biosynthetic enzymes. The review will help us to advance our understanding of saponin biosynthesis and aid in the development of molecular and biotechnological tools for the systematic optimization of metabolic fluxes, in order to produce the desired saponins in pulses.

Seed development in angiosperms produces three genetically and developmentally distinct sub-compartments: the embryo, endosperm, and seed coat. The maternally derived seed coat protects the embryo and interacts closely with the external environment especially during germination and seedling establishment. Seed coat is a key contributor to seed composition and an important determinant of nutritional value for humans and livestock. In this review, we examined pea crop productivity through the lens of the seed coat, its contribution to several valued nutritional traits of the pea crop, and its potential as a breeding target. Key discoveries made in advancing the knowledge base for sensing and transmission of external signals, the architecture and chemistry of the pea seed coat, and relevant insights from other important legumes were discussed. Furthermore, for selected seed coat traits, known mechanisms of genetic regulation and efforts to modulate these mechanisms to facilitate composition and productivity improvements in pea were discussed, alongside opportunities to support the continued development and improvement of this underutilized crop. This review describes the most important features of seed coat development in legumes and highlights the key roles played by the seed coat in pea seed development, with a focus on advances made in the genetic and molecular characterization of pea and other legumes and the potential of this key seed tissue for targeted improvement and crop optimization.

Starch is the most abundant storage carbohydrate and a major component in pea seeds, accounting for about 50% of dry seed weight. As a by-product of pea protein processing, current uses for pea starch are limited to low-value, commodity markets. The globally growing demand for pea protein poses a great challenge for the pea fractionation industry to develop new markets for starch valorization. However, there exist gaps in our understanding of the genetic mechanism underlying starch metabolism, and its relationship with physicochemical and functional properties, which is a prerequisite for targeted tailoring functionality and innovative applications of starch. This review outlines the understanding of starch metabolism with a particular focus on peas and highlights the knowledge of pea starch granule structure and its relationship with functional properties, and industrial applications. Using the currently available pea genetics and genomics knowledge and breakthroughs in omics technologies, we discuss the perspectives and possible avenues to advance our understanding of starch metabolism in peas at an unprecedented level, to ultimately enable the molecular design of multi-functional native pea starch and to create value-added utilization.

Grain protein concentration (GPC) is an important trait in durum cultivar development as a major determinant of the nutritional value of grain and end-use product quality. However, it is challenging to simultaneously select both GPC and grain yield (GY) due to the negative correlation between them. To characterize quantitative trait loci (QTL) for GPC and understand the genetic relationship between GPC and GY in Canadian durum wheat, we performed both traditional and conditional QTL mapping using a doubled haploid (DH) population of 162 lines derived from Pelissier × Strongfield. The population was grown in the field over 5 years and GPC was measured. QTL contributing to GPC were detected on chromosome 1B, 2B, 3A, 5B, 7A, and 7B using traditional mapping. One major QTL on 3A (QGpc.spa-3A.3) was consistently detected over 3 years accounting for 9.4–18.1% of the phenotypic variance, with the favorable allele derived from Pelissier. Another major QTL on 7A (QGpc.spa-7A) detected in 3 years explained 6.9–14.8% of the phenotypic variance, with the beneficial allele derived from Strongfield. Comparison of the QTL described here with the results previously reported led to the identification of one novel major QTL on 3A (QGpc.spa-3A.3) and five novel minor QTL on 1B, 2B and 3A. Four QTL were common between traditional and conditional mapping, with QGpc.spa-3A.3 and QGpc.spa-7A detected in multiple environments. The QTL identified by conditional mapping were independent or partially independent of GY, making them of great importance for development of high GPC and high yielding durum.

Gluten strength is one of the factors that determine the end-use quality of durum wheat and is an important breeding target for this crop. To characterize the quantitative trait loci (QTL) controlling gluten strength in Canadian durum wheat cultivars, a population of 162 doubled haploid (DH) lines segregating for gluten strength and derived from cv. Pelissier × cv. Strongfield was used in this study. The DH lines, parents, and controls were grown in 3 years and two seeding dates in each year and gluten strength of grain samples was measured by sodium dodecyl sulfate (SDS)-sedimentation volume (SV). With a genetic map created by genotyping the DH lines using the Illumina Infinium iSelect Wheat 90K SNP (single nucleotide polymorphism) chip, QTL contributing to gluten strength were detected on chromosome 1A, 1B, 2B, and 3A. Two major and stable QTL detected on chromosome 1A (QGlu.spa-1A) and 1B (QGlu.spa-1B.1) explaining 13.7–18.7% and 25.4–40.1% of the gluten strength variability respectively were consistently detected over 3 years, with the trait increasing alleles derived from Strongfield. Putative candidate genes underlying the major QTL were identified. Two novel minor QTL (QGlu.spa-3A.1 and QGlu.spa-3A.2) with the trait increasing allele derived from Pelissier were mapped on chromosome 3A explaining up to 8.9% of the phenotypic variance; another three minor QTL (QGlu.spa-2B.1, QGlu.spa-2B.2, and QGlu.spa-2B.3) located on chromosome 2B explained up to 8.7% of the phenotypic variance with the trait increasing allele derived from Pelissier. QGlu.spa-2B.1 is a new QTL and has not been reported in the literature. Multi-environment analysis revealed genetic (QTL) × environment interaction due to the difference of effect in magnitude rather than the direction of the QTL. Eleven pairs of digenic epistatic QTL were identified, with an epistatic effect between the two major QTL of QGlu.spa-1A and QGlu.spa-1B.1 detected in four out of six environments. The peak SNPs and SNPs flanking the QTL interval of QGlu.spa-1A and QGlu.spa-1B.1 were converted to Kompetitive Allele Specific PCR (KASP) markers, which can be deployed in marker-assisted breeding to increase the efficiency and accuracy of phenotypic selection for gluten strength in durum wheat. The QTL that were expressed consistently across environments are of great importance to maintain the gluten strength of Canadian durum wheat to current market standards during the genetic improvement.

Background: The Arabidopsis microRNA156 (miR156) regulates 11 members of the SQUAMOSA PROMOTER BINDING PROTEIN LIKE (SPL) family by base pairing to complementary target mRNAs. Each SPL gene further regulates a set of other genes; thus, miR156 controls numerous genes through a complex gene regulation network. Increased axillary branching occurs in transgenic Arabidopsis overexpressing miR156b, similar to that observed in loss-of-function max3 and max4 mutants with lesions in carotenoid cleavage dioxygenases. Arabidopsis miR156b was found to enhance carotenoid levels and reproductive shoot branching when expressed in Brassica napus, suggesting a link between miR156b expression and carotenoid metabolism. However, details of the miR156 regulatory network of SPL genes related to carotenoid metabolism are not known. Results: In this study, an Arabidopsis T-DNA enhancer mutant, sk156, was identified due to its altered branching and trichome morphology and increased seed carotenoid levels compared to wild type (WT) ecovar Columbia. Enhanced miR156b expression due to the 35S enhancers present on the T-DNA insert was responsible for these phenotypes. Constitutive and leaf primodium-specific expression of a miR156-insensitive (mutated) SPL15 (SPL15m) largely restored WT seed carotenoid levels and plant morphology when expressed in sk156. The Arabidopsis native miR156-sensitive SPL15 (SPL15n) and SPL15m driven by a native SPL15 promoter did not restore the WT phenotype in sk156. Our findings suggest that SPL15 function is somewhat redundant with other SPL family members, which collectively affect plant phenotypes. Moreover, substantially decreased miR156b transcript levels in sk156 expressing SPL15m, together with the presence of multiple repeats of SPL-binding GTAC core sequence close to the miR156b transcription start site, suggested feedback regulation of miR156b expression by SPL15. This was supported by the demonstration of specific in vitro interaction between DNA-binding SBP domain of SPL15 and the proximal promoter sequence of miR156b. Conclusions: Enhanced miR156b expression in sk156 leads to the mutant phenotype including carotenoid levels in the seed through suppression of SPL15 and other SPL target genes. Moreover, SPL15 has a regulatory role not only for downstream components, but also for its own upstream regulator miR156b.