Genomic asymmetry of the Brassica napus seed: epigenetic contributions of DNA methylation and small RNAs to subgenome bias.

Polyploidy is a persistent phenomenon in angiosperm genome evolution that is hypothesized to have contributed to the diversity of extant flowering plants. Brassica napus, one of the world's most important angiosperm oilseed species, originated from the interspecific hybridization of Brassica rapa (An ) and Brassica oleracea (Cn ). While the trends of genome dominance in transcriptomics are beginning to emerge, less is known about the epigenetic and small RNA landscapes in polyploids during reproductive development. The seed is the pivotal developmental transition into the new sporophytic generation, and experiences substantial epigenetic modifications over time. Here, we investigated the prevalence of bias in the contexts of DNA methylation and small interfering (si)RNA profiles in both subgenomes (An and Cn ), as well as the ancestral fractionated genomes across B. napus seed development. We report ubiquitous Cn subgenome bias of siRNA expression and cytosine methylation, with DNA methylation being particularly abundant on gene promoters in the Cn subgenome. Further, we provide evidence that siRNA transcriptional patterns were conserved within the ancestral triplicated subgenomes of B. napus, but not across the An and Cn subgenomes. We discuss how methylation patterns in the B. napus seed relate to genes, promoter regions, siRNA loci and transposable elements through the lens of genome fractionation and polyploidization. Taken together we provide evidence for epigenetic regulation selectively silencing the Cn subgenome during seed development, and explore the impact of genome fractionation on the epigenetic components of the B. napus seed.

Alleviating salinity stress in canola (Brassica napus L.) through exogenous application of salicylic acid.

Canola, a vital oilseed crop, is grown globally for food and biodiesel. With the enormous demand for growing various crops, the utilization of agriculturally marginal lands is emerging as an attractive alternative, including brackish-saline transitional lands. Salinity is a major abiotic stress limiting growth and productivity of most crops, and causing food insecurity. Salicylic acid (SA), a small-molecule phenolic compound, is an essential plant defense phytohormone that promotes immunity against pathogens. Recently, several studies have reported that SA was able to improve plant resilience to withstand high salinity. For this purpose, a pot experiment was carried out to ameliorate the negative effects of sodium chloride (NaCl) on canola plants through foliar application of SA. Two canola varieties Faisal (V1) and Super (V2) were assessed for their growth performance during exposure to high salinity i.e. 0 mM NaCl (control) and 200 mM NaCl. Three levels of SA (0, 10, and 20 mM) were applied through foliar spray. The experimental design used for this study was completely randomized design (CRD) with three replicates. The salt stress reduced the shoot and root fresh weights up to 50.3% and 47% respectively. In addition, foliar chlorophyll a and b contents decreased up to 61-65%. Meanwhile, SA treatment diminished the negative effects of salinity and enhanced the shoot fresh weight (49.5%), root dry weight (70%), chl. a (36%) and chl. b (67%). Plants treated with SA showed an increased levels of both enzymatic i.e. (superoxide dismutase (27%), peroxidase (16%) and catalase (34%)) and non-enzymatic antioxidants i.e. total soluble protein (20%), total soluble sugar (17%), total phenolic (22%) flavonoids (19%), anthocyanin (23%), and endogenous ascorbic acid (23%). Application of SA also increased the levels of osmolytes i.e. glycine betaine (31%) and total free proline (24%). Salinity increased the concentration of Na+ ions and concomitantly decreased the K+ and Ca2+ absorption in canola plants. Overall, the foliar treatments of SA were quite effective in reducing the negative effects of salinity. By comparing both varieties of canola, it was observed that variety V2 (Super) grew better than variety V1 (Faisal). Interestingly, 20 mM foliar application of SA proved to be effective in ameliorating the negative effects of high salinity in canola plants.

A CRISPR/Cas9-based vector system enables the fast breeding of selection-marker-free canola with Rcr1-rendered clubroot resistance.

Breeding for disease resistance in major crops is of crucial importance for global food security and sustainability. However, common biotechnologies such as traditional transgenesis or genome editing do not provide an ideal solution, whereas transgenic crops free of selection markers such as cisgenic/intragenic crops might be suitable. In this study, after cloning and functional verification of the Rcr1 gene for resistance to clubroot (Plasmodiophora brassicae), we confirmed that the genes Rcr1, Rcr2, Rcr4, and CRa from Brassica rapa crops and the resistance gene from B. napus oilseed rape cv. 'Mendel' on chromosome A03 were identical in their coding regions. We also determined that Rcr1 has a wide distribution in Brassica breeding materials and renders potent resistance against multiple representative clubroot strains in Canada. We then modified a CRISPR/Cas9-based cisgenic vector system and found that it enabled the fast breeding of selection-marker-free transgenic crops with add-on traits, with selection-marker-free canola (B. napus) germplasms with Rcr1-rendered stable resistance to clubroot disease being successfully developed within 2 years. In the B. napus background, the intragenic vector system was able to remove unwanted residue sequences from the final product with high editing efficiency, and off-target mutations were not detected. Our study demonstrates the potential of applying this breeding strategy to other crops that can be transformed by Agrobacterium. Following the streamlined working procedure, intragenic germplasms can be developed within two generations, which could significantly reduce the breeding time and labor compared to traditional introgression whilst still achieving comparable or even better breeding results.

Fluorescence Spectroscopy Based Characterization of Flaxseed Oil.

Fluorescence spectroscopy has been employed for the compositional analysis of flaxseed oil, detection of its adulteration and investigation of the thermal effects on its molecular composition. Excitation wavelengths from 320 to 420 nm have been used to explore the valued ingredients in flaxseed oil. The emission bands of flaxseed oil centred at 390, 414, 441, 475, 515 and 673/720 nm represent vitamin K, isomers of vitamin E, carotenoids and chlorophylls, which can be used as a marker for quality analysis. Due to its high quality, it is highly prone to adulteration and in this study, detection of its adulteration with canola oil is demonstrated by applying principal component analysis. Moreover, the effects of temperature on the molecular composition of cold pressed flaxseed oil has been explored by heating them at cooking temperatures of 100, 110, 120, 130, 140, 150, 160, 170 and 180 °C, each for 30 min. On heating, the deterioration of vitamin E, carotenoids and chlorophylls occurred with an increase in the oxidation products. However, it was found that up to 140 °C, flaxseed oil retains much of its natural composition whereas up to 180 oC, it loses much of its valuable ingredients along with increase of oxidized products.

Added γ-oryzanol boosted anti-inflammatory effects of canola oil in adult subjects with type 2 diabetes: a randomized controlled clinical trial.

PURPOSE: This study was conducted to examine the effects of daily intake of γ-oryzanol (ORZ)-fortified canola oil, as compared with plain canola and sunflower oils, on certain inflammatory and oxidative stress biomarkers in adult subjects with Type 2 Diabetes (T2D). METHODS: We randomly allocated 92 adult subjects with T2D from both sexes to one of the following groups to receive: (a) ORZ-fortified canola oil (ORZO; n1 = 30); (b) unfortified canola oil (CANO; n2 = 32); or (c) sunflower oil (SUFO; n3 = 30) for 12 weeks. Dietary and laboratory evaluations were performed initially and finally. RESULTS: Serum hs-CRP concentrations significantly decreased in ORZO group (from 3.1 ± 0.2 to 1.2 ± 0.2 mg/L), as compared with CANO (p = 0.003) and SUFO (p < 0.001) groups. Serum IL-6 significantly decreased just in ORZO (- 22.8%, p = 0.042) and CANO groups (- 19.8%, p = 0.038). However, the between-group differences were not significant. Serum IL-1ß slightly decreased in ORZO (- 28.1%, p = 0.11) and increased in SUFO (+ 20.6%, p = 0.079) but between-group difference was statistically significant (p = 0.017). Serum IFN-γ concentrations decreased significantly only in ORZO (from 3.3 ± 0.08 to 2.9 ± 0.21 IU/mL, p = 0.044). Salivary IgA concentrations increased significantly in all three intervention groups. Notwithstanding, only the difference between ORZO and CANO groups was statistically significant (p = 0.042). Similarly, circulating malondialdehyde concentrations significantly decreased in all three groups but with no between-group significant difference. CONCLUSIONS: Daily consumption of ORZ-fortified canola oil, compared with unfortified canola and sunflower oils, for 12 weeks resulted in boosting of certain anti-inflammatory effects of canola oil. These findings may have preventive implications for both clinicians and policy makers. This clinical trial was registered at clinicaltrials.gov (03.08.2022; NCT05271045).

Milk production of cows grazing pasture supplemented with grain mixes containing canola meal or corn grain or both over the first 100 days of lactation.

Grain mixes varying in proportions of wheat grain, barley grain, canola meal, and corn grain were fed to grazing dairy cows in early lactation to determine the contribution of canola meal and corn grain to milk yield, BW, BCS, eating behavior, and blood serum metabolite concentrations. The experiment used 80 multiparous, seasonally calving Holstein-Friesian dairy cows during the first 100 d of lactation, the treatment period, and over the subsequent carryover period of 100 d, during which all cows were fed a common diet. Cows were divided into 4 cohorts (blocks) based on calving date and within each cohort, 5 cows were randomly allocated to each of the 4 treatments. Dietary treatments included disc-milled grain mixes comprising (on a DM basis) (1) a control treatment of wheat (25%) and barley (75%); (2) wheat (25%), barley (50%), and canola meal (25%); (3) wheat (25%), barley (50%), and corn (25%), and (4) wheat (25%), barley (25%), canola meal (25%), and corn (25%). Treatment diets were introduced at 19 DIM ± 4.7 d, which included a 7-d adaptation period and were applied up until 100 DIM. Each grain mix was fed at 9 kg of DM/cow per day, offered twice daily, in equal proportions in the parlor at milking times. In addition to the grain mix, all cows grazed perennial ryegrass pasture at a daily allowance of ∼35 kg of DM/cow per day (measured to ground level). Results were analyzed in terms of corn and canola presence or absence in the diet. Including canola meal in grain mixes increased grain intake and pasture intake by 0.6 and 2.1 kg of DM/cow per day, respectively, resulting in an increased milk yield of 2.6 kg/cow per day during the first 100 d of lactation. Including canola meal also increased yields of milk fat and protein, and concentrations of milk fat, as well as increasing mean BW and BCS over the 100 d. The inclusion of canola meal in the grain mixes also resulted in greater blood serum BHB and urea concentrations, compared with feeding grain mixes that did not contain canola meal. The inclusion of corn grain provided no milk production benefits and did not change BW, BCS, or any feeding behavior variables. There were no carryover effects on milk production from either canola meal or corn grain after the treatment period. In summary, the results demonstrate that the provision of canola meal in grain mixes can improve milk production and increase mean BCS. Further, there are no benefits to milk yield when a proportion of barley is substituted for corn, in a wheat and barley grain mix fed to grazing dairy cows in early lactation. However, these results are dependent on the level of inclusion and the feeding system employed.

Effects of protein and forage source on performance, and splanchnic and mammary net fluxes of nutrients in lactating dairy cows.

This study was conducted to determine if the decreased MP supply predicted by the NRC (2001) when canola meal (CM) substitutes soybean meal (SBM) was supported by direct measurement of net portal absorption of AA or energy-yielding nutrients, plus the impact of the type of forage in CM-based rations. Nine Holstein cows with indwelling catheters in splanchnic blood vessels, 8 also with a ruminal cannula were used to examine the effects of protein source in corn silage-based diets, comparing SBM versus CM, and forage source in CM-based diets, comparing corn versus grass silage. The cows were allocated to a triple 3 × 3 Latin square design with 21-d periods. The 3 experimental diets, formulated to be isoenergetic and isonitrogenous, were based on: 1) SBM and corn silage (SoyCorn); 2) CM and corn silage (CanCorn) and 3) CM and cool-season grass silage (CanGrass). Averages of intake, milk yield and milk composition of the last 3 d of each period were used for statistical analyses. On d 21 of each period, 6 sets of arterial, portal, hepatic and mammary blood samples and 2 ruminal fluid samples were collected. On d 12 of period 2, the protein sources were incubated in nylon bags to determine 16h-ruminal disappearance of DM and N and to obtain 16-h residues. Finally, 5 d after the completion of the Latin square design, the mobile bag technique was used to determine DM and N intestinal disappearance of the 16-h residues of SBM and CM. Pre-planned contrasts were used to compare the effect of the protein source in cows fed corn silage, i.e., SoyCorn versus CanCorn, and the effect of forage in cows fed CM, i.e., CanCorn versus CanGrass. Data of the cow without a rumen canula could not be used because of health problem. In corn silage-based diets, substitution of SBM by CM tended to increase milk (6%) and milk fat (7%) yields. The 8% higher ruminal N disappearance and the 19% decreased MP supply from RUP predicted by NRC (2001) were not supported by the 25% decrease in ruminal ammonia concentration, similar net portal absorption of AA (except 22% higher for Met), and the 14% decrease in urea hepatic removal when CM substituted SBM. Ruminal incubation of CM in nylon bags does not appear suitable for adequate determination of the rumen by-pass of a protein source like CM. Inclusion of grass silage rather than corn silage in CM-based diets tended to increase milk (6%) and increased milk lactose (8%) yields. Neither protein nor forage source resulted in variations of metabolism of energy-yielding nutrients that could explain observed increments in cow performance. The present study indicates no decreased AA availability when CM substitutes SBM. Therefore, substitution of SBM by CM in diets based on corn silage and CM in corn- or grass silage-diets can be used successfully in high producing dairy cows.

A Hydroponic-Based Bioassay to Facilitate Plasmodiophora brassicae Phenotyping.

Clubroot, caused by the obligate parasite Plasmodiophora brassicae, is one of the most devastating diseases affecting the canola/oilseed rape (Brassica napus) industry worldwide. Currently, the planting of clubroot-resistant (CR) cultivars is the most effective strategy used to restrict the spread and the economic losses linked to the disease. However, virulent P. brassicae isolates have been able to infect many of the currently available CR cultivars, and the options to manage the disease are becoming limited. Another challenge has been achieving consistency in evaluating host reactions to P. brassicae infection, with most bioassays conducted in soil and/or potting medium, which requires significant space and can be labor intensive. Visual scoring of clubroot symptom development can also be influenced by user bias. Here, we have developed a hydroponic bioassay using well-characterized P. brassicae single-spore isolates representative of clubroot virulence in Canada, as well as field isolates from three Canadian provinces in combination with canola inbred homozygous lines carrying resistance genetics representative of CR cultivars available to growers in Canada. To improve the efficiency and consistency of disease assessment, symptom severity scores were compared with clubroot evaluations based on the scanned root area. According to the results, this bioassay offers a reliable, less expensive, and reproducible option to evaluate P. brassicae virulence, as well as to identify which canola resistance profile(s) may be effective against particular isolates. This bioassay will contribute to the breeding of new CR canola cultivars and the identification of virulence genes in P. brassicae that could trigger resistance and that have been very elusive to this day.[Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Diversity and Pathogenicity of Fusarium Root Rot Fungi from Canola (Brassica napus) in Alberta, Canada.

Root rot disease poses a significant threat to canola (Brassica napus), underscoring the need for a comprehensive understanding of its causal agents for more effective disease mitigation. The composition and diversity of fungal pathogens associated with root rot of canola in Alberta, Canada, were evaluated from plant tissue samples collected in 2021 and 2022. The study revealed Fusarium spp. as the predominant pathogens found in almost all surveyed fields. Fusarium avenaceum, F. redolens, and F. solani were among the most frequently recovered species. Greenhouse trials confirmed their pathogenicity, with F. avenaceum and F. sporotrichioides found to be particularly aggressive. Additionally, F. sporotrichioides and F. commune were identified for the first time as canola root rot pathogens. Inoculation with isolates of most species resulted in significant reductions in seedling emergence, plant height, and shoot and root dry weights. Analysis of translation elongation factor 1-α (TEF-1α) and internal transcribed spacer (ITS) sequences confirmed the identity of the Fusarium spp., while concatenating the ITS and TEF-1α sequences enabled improved species differentiation. Geographic and year effects did not influence fungal diversity or aggressiveness, as determined by principal component analysis. This study emphasized the high diversity and impact of Fusarium spp. in causing canola root rot.

Catalytic Supercritical Water Gasification of Canola Straw with Promoted and Supported Nickel-Based Catalysts.

Lignocellulosic biomass such as canola straw is produced as low-value residue from the canola processing industry. Its high cellulose and hemicellulose content makes it a suitable candidate for the production of hydrogen via supercritical water gasification. However, supercritical water gasification of lignocellulosic biomass such as canola straw suffers from low hydrogen yield, hydrogen selectivity, and conversion efficiencies. Cost-effective and sustainable catalysts with high catalytic activity for supercritical water gasification are increasingly becoming a focal point of interest. In this research study, novel wet-impregnated nickel-based catalysts supported on carbon-negative hydrochar obtained from hydrothermal liquefaction (HTL-HC) and hydrothermal carbonization (HTC-HC) of canola straw, along with other nickel-supported catalysts such as Ni/Al2O3, Ni/ZrO2, Ni/CNT, and Ni/AC, were synthesized for gasification of canola straw on previously optimized reaction conditions of 500 °C, 60 min, 10 wt%, and 23-25 MPa. The order of hydrogen yield for the six supports was (10.5 mmol/g) Ni/ZrO2 > (9.9 mmol/g) Ni/Al2O3 > (9.1 mmol/g) Ni/HTL-HC > (8.8 mmol/g) Ni/HTC-HC > (7.7 mmol/g) Ni/AC > (6.8 mmol/g) Ni/CNT, compared to 8.1 mmol/g for the non-catalytic run. The most suitable Ni/ZrO2 catalyst was further modified using promotors such as K, Zn, and Ce, and the performance of the promoted Ni/ZrO2 catalysts was evaluated. Ni-Ce/ZrO2 showed the highest hydrogen yield of 12.9 mmol/g, followed by 12.0 mmol/g for Ni-Zn/ZrO2 and 11.6 mmol/g for Ni-K/ZrO2. The most suitable Ni-Ce/ZrO2 catalysts also demonstrated high stability over their repeated use. The superior performance of the Ni-Ce/ZrO2 was due to its high nickel dispersion, resilience to sintering, high thermal stability, and oxygen storage capabilities to minimize coke deposition.

Apparent ileal amino acid digestibility, gut morphometrics, and gene expression of peptide and amino acid transporters in broiler chickens fed low-crude-protein diets supplemented with crystalline amino acids with soybean meal, canola meal, or corn DDGS as protein feedstuffs.

BACKGROUND: We investigated the impact of using canola meal (CM) or corn distillers dried grain soluble (cDDGS) in place of soybean meal (SBM) in low-crude-protein diets supplemented with amino acids (AA) on AA digestibility, gut morphometrics, and AA transporter genes in broiler chicken. On day 0, 540 Cobb 500 male broilers were allocated to six diets in 36-floor pens. The positive control (PC) was a corn-SBM diet with adequate crude protein (CP). The CP level of negative control (NC) was decreased by 45 and 40 g kg-1 relative to PC for grower and finisher phases, respectively. The subsequent two diets had the same CP levels as NC but with cDDGS added at 50 or 125 g kg-1. The last two diets had the same CP as NC but with CM added at 50 or 100 g kg-1. RESULTS: Dietary CP reduction in corn-SBM diets increased (P < 0.05) the digestibility of Lys (88.5%), Met (90.7%), Thr (77.4%), Cys (80.7%), and Gly (84.7%). Increasing levels of cDDGS linearly decreased (P < 0.05) the digestibility of Asp, Cys, Glu, and Ser, whereas increasing CM level linearly decreased (P < 0.05) the digestibility of Cys, Pro, and Ser. The CP reduction in corn-SBM diets produced downward expression of peptide transporter1 and decreased (P < 0.05) absolute pancreas and ileum weight and length of jejunum and ileum. CONCLUSIONS: Partial replacement of SBM with alternative protein feedstuffs (cDDGS or CM) in low-CP diets had minimal effects on AA digestibility and mRNA levels of peptides and AA transporters. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Evaluation of growth, nutrient absorption, body composition and blood indices under dietary exposure of iron oxide nanoparticles in Common carp (Cyprinus carpio).

The bioavailability, small size and direct absorption in the blood, make nanoparticles (NPs) a remarkable feed additive in the aquaculture industry. Therefore, dietary iron oxide nanoparticles (Fe2 O3 -NPs) were used to examine their effects on growth, nutrient absorption, body composition and blood indices in Cyprinus carpio (Common carp) fingerlings. Healthy C. carpio fingerlings (n = 270) were fed with six canola meal based experimental diets (D1-control, D2, D3, D4, D5, D6) supplemented with 0, 10, 20, 30, 40 and 50 mg/kg Fe2 O3 -NPs respectively. A total of 15 fingerlings (average initial weight 5.51 ± 0.04 g/fish) were kept in triplicates for 70 days. The results indicated that maximum growth performance, apparent digestibility coefficient, body composition and haematological parameters were observed in 40 mg/kg Fe2 O3 -NPs supplementation. All the experimental diets were significantly improved (p < 0.05) in all the above parameters than control diet. In the present research, the recommended dosage of Fe2 O3 -NPs as dietary supplement is 40 mg/kg for improving the growth, nutrient absorption, body composition and haematological indices in C. carpio fingerlings. Hence, this study demonstrates the potential of NPs to improve the health of fish.

Characterization of ruminal degradation, intestinal digestion and total true nutrient supply to dairy cows from feedstocks and coproducts from Canola bio-oil processing: Impact by source origin.

The objective of this study was to characterize ruminal degradation, intestinal digestion and total true nutrient supply to dairy cows from canola feedstock (canola seeds) and coproducts (meal and pellets) from bio-oil processing which were impacted by source origin. The feedstocks and coproducts (mash, pellet) were randomly collected from five different bio-oil processing plants with five different batches of samples in each bio-processing plant in Canada (CA) and China (CH). In situ rumen degradation kinetics were determined using four fistulated Holstein cows with incubation times at 0, 2, 4, 8, 12, 24 and 48 h. Intestinal digestions were determined using the three-step in vitro method with preincubation at 12 h. The DVE/OEB and National Research Council systems were applied to evaluate the truly absorbable nutrient supply to dairy cows and feed milk values (FMVs). The results showed that in situ undegradable fractions (U) (p = 0.025) were higher in CA meals, and potentially degradable fraction of D was higher (p = 0.016) in CH meals. CH meals had higher total digestible dry matter (TDDM, p = 0.018) and intestinal digestibility of protein (dIDP, p = 0.016). Canola meals from CA had lower MREE (microbial protein synthesized in the rumen based on available rumen degradable protein; p = 0.011) and DVME (rumen synthesized microbial protein digested in the small intestine; p = 0.011) and had higher ECP (endogenous protein in the small intestine, p = 0.001) and absorbed endogenous crude protein (truly absorbed ECP in the small intestine) than CH (p = 0.001). The FMV evaluated based on the metabolic protein and net energy showed no differences between CA and CH in both coproducts and feedstocks.

Gene expression profiling reveals transcription factor networks and subgenome bias during Brassica napus seed development.

We profiled the global gene expression landscape across the reproductive lifecycle of Brassica napus. Comparative analysis of this nascent amphidiploid revealed the contribution of each subgenome to plant reproduction. Whole-genome transcription factor networks identified BZIP11 as a transcriptional regulator of early B. napus seed development. Knockdown of BZIP11 using RNA interference resulted in a similar reduction in gene activity of predicted gene targets, and a reproductive-lethal phenotype. Global mRNA profiling revealed lower accumulation of Cn subgenome transcripts relative to the An subgenome. Subgenome-specific transcription factor networks identified distinct transcription factor families enriched in each of the An and Cn subgenomes early in seed development. Analysis of laser-microdissected seed subregions further reveal subgenome expression dynamics in the embryo, endosperm and seed coat of early stage seeds. Transcription factors predicted to be regulators encoded by the An subgenome are expressed primarily in the seed coat, whereas regulators encoded by the Cn subgenome were expressed primarily in the embryo. Data suggest subgenome bias are characteristic features of the B. napus seed throughout development, and that such bias might not be universal across the embryo, endosperm and seed coat of the developing seed. Transcriptional networks spanning both the An and Cn genomes of the whole B. napus seed can identify valuable targets for seed development research and that -omics level approaches to studying gene regulation in B. napus can benefit from both broad and high-resolution analyses.

Brassica napus Haploid and Double Haploid Production and Its Latest Applications.

Rapeseed is one of the most important oil crops in the world. Increasing demand for oil and limited agronomic capabilities of present-day rapeseed result in the need for rapid development of new, superior cultivars. Double haploid (DH) technology is a fast and convenient approach in plant breeding as well as genetic research. Brassica napus is considered a model species for DH production based on microspore embryogenesis; however, the molecular mechanisms underlying microspore reprogramming are still vague. It is known that morphological changes are accompanied by gene and protein expression patterns, alongside carbohydrate and lipid metabolism. Novel, more efficient methods for DH rapeseed production have been reported. This review covers new findings and advances in Brassica napus DH production as well as the latest reports related to agronomically important traits in molecular studies employing the double haploid rapeseed lines.

Targeted editing of multiple homologues of GTR1 and GTR2 genes provides the ideal low-seed, high-leaf glucosinolate oilseed mustard with uncompromised defence and yield.

Glucosinolate content in the two major oilseed Brassica crops-rapeseed and mustard has been reduced to the globally accepted Canola quality level (<30 µmoles/g of seed dry weight, DW), making the protein-rich seed meal useful as animal feed. However, the overall lower glucosinolate content in seeds as well as in the other parts of such plants renders them vulnerable to biotic challenges. We report CRISPR/Cas9-based editing of glucosinolate transporter (GTR) family genes in mustard (Brassica juncea) to develop ideal lines with the desired low seed glucosinolate content (SGC) while maintaining high glucosinolate levels in the other plant parts for uncompromised plant defence. Use of three gRNAs provided highly efficient and precise editing of four BjuGTR1 and six BjuGTR2 homologues leading to a reduction of SGC from 146.09 µmoles/g DW to as low as 6.21 µmoles/g DW. Detailed analysis of the GTR-edited lines showed higher accumulation and distributional changes of glucosinolates in the foliar parts. However, the changes did not affect the plant defence and yield parameters. When tested against the pathogen Sclerotinia sclerotiorum and generalist pest Spodoptera litura, the GTR-edited lines displayed a defence response at par or better than that of the wild-type line. The GTR-edited lines were equivalent to the wild-type line for various seed yield and seed quality traits. Our results demonstrate that simultaneous editing of multiple GTR1 and GTR2 homologues in mustard can provide the desired low-seed, high-leaf glucosinolate lines with an uncompromised defence and yield.

The transcription factor BnaA9.WRKY47 coordinates leaf senescence and nitrogen remobilization in Brassica napus.

Nitrogen (N) is an essential macronutrient for plants, and its remobilization is key for adaptation to deficiency stress. However, there is limited understanding of the regulatory mechanisms of N remobilization in the important crop species Brassica napus (oilseed rape). Here, we report the identification of a transcription factor, BnaA9.WRKY47, that is induced by N starvation in a canola variety. At the seedling stage, BnaA9.WRKY47-overexpressing (OE) lines displayed earlier senescence of older leaves and preferential growth of juvenile leaves compared to the wild type under N starvation. At the field scale, the seed yield was significantly increased in the BnaA9.WRKY47-OE lines compared with the wild type when grown under N deficiency conditions and, conversely, it was reduced in BnaA9.WRKY47-knockout mutants. Biochemical analyses demonstrated that BnaA9.WRKY47 directly activates BnaC7.SGR1 to accelerate senescence of older leaves. In line with leaf senescence, the concentration of amino acids in the older leaves of the OE lines was elevated, and the proportion of plant N that they contained was reduced. This was associated with BnaA9.WRKY47 activating the amino acid permease BnaA9.AAP1 and the nitrate transporter BnaA2.NRT1.7. Thus, the expression of BnaA9.WRKY47 efficiently facilitated N remobilization from older to younger leaves or to seeds. Taken together, our results demonstrate that BnaA9.WRKY47 up-regulates the expression of BnaC7.SGR1, BnaA2.NRT1.7, and BnaA9AAP1, thus promoting the remobilization of N in B. napus under starvation conditions.

The Programming Effect of Plant-Based DHA, Along with Equivalent AA, on Immune System and Oral Tolerance Development in Six-Week Allergy Prone BALB/c Pups.

BACKGROUND: Docosahexaenoic acid (DHA) and arachidonic acid (AA) on oral tolerance (OT) development in allergy-prone infants is less known. OBJECTIVES: We aim to determine the effects of early life DHA supplementation (1% of total fat, from novel canola oil), along with AA, on OT toward ovalbumin (ova, egg protein) in allergy-prone BALB/c pups at 6-wk. METHODS: Breastfeeding dams (n ≥ 10/diet) were fed DHA+AA (1% DHA, 1% AA wt/wt of total fat) or control (0% DHA, 0% AA) suckling period diet (SPD) during which pups consumed dam's milk. At 3-wk, pups from each SPD group were assigned to either the control or DHA+AA weaning diet. For OT, pups from each diet group were either orally fed ova or placebo daily from 21-25 d. Systemic immunization to ova was induced through intraperitoneal injections before euthanizing 6-wk pups. Ova-specific immunoglobulin (ova-Ig) and splenocytes ex-vivo cytokine response to different stimuli were analyzed using a 3-factor analysis of variance. RESULTS: OT-induced suppression was seen in ova-stimulated splenocyte ex-vivo response, where ova-tolerized pups showed significantly lower total immunoglobulin (Ig)G, IgG1, interleukin (IL)-2 and IL-6 production than sucrose (placebo) pups. DHA+AA SPD was associated with 3 times lower plasma concentrations of ova-IgE (P = 0.03) than controls. DHA+AA weaning diet resulted in lower T helper type-2 cytokines (IL-4 and IL-6) with ova stimulation than controls, which may benefit OT. DHA+AA SPD resulted in significantly higher T cell cytokine response [IL-2, interferon-gamma, (IFNγ) and IL-1ß] to anti-CD3/CD28 stimulation than controls. The splenocytes stimulated with lipopolysaccharide produced lower inflammatory cytokines (IFNγ, tumor necrosis factor-alpha, IL-6, and C-X-C motif ligand 1), which may be because of lower CD11b+CD68+ splenocytes proportion in pups from DHA+AA SPD than control (all P < 0.05). CONCLUSIONS: DHA and AA in early life may influence OT in allergy-prone BALB/c mouse offspring, as they effectively promote T helper type-1 immune responses.

Research progress and future study on physicochemical, nutritional, and structural characteristics of canola and rapeseed feedstocks and co-products from bio-oil processing and nutrient modeling evaluation methods.

This article aims to review research progress and provide future study on physicochemical, nutritional, and molecular structural characteristics of canola and rapeseed feedstocks and co-products from bio-oil processing and nutrient modeling evaluation methods. The review includes Canola oil seed production, utilization and features; Rapeseed oil seed production and canola oil seed import in China; Bio-processing, co-products and conventional evaluation methods; Modeling methods for evaluation of truly absorbed protein supply from canola feedstock and co-products. The article provides our current research in feedstocks and co-products from bio-oil processing which include Characterization of chemical and nutrient profiles and ruminal degradation and intestinal digestion; Revealing intrinsic molecular structures and relationship between the molecular structure spectra features and nutrient supply from feedstocks and co-products using advanced vibrational molecular spectroscopy technique. The study focused on advanced vibrational molecular spectroscopy which can be used as a fast tool to study molecular structure features of feedstock and co-products from bio-oil processing. The article also provides future in depth study areas. This review provides an insight as how to use advanced vibrational molecular spectroscopy for in-depth analysis of the relationship between molecular structure spectral feature and nutrition delivery from canola feedstocks and co-products from bio-oil processing.

Quality Analysis of Canola and Mustard Oil Using Fluorescence Spectroscopy.

The potential of Fluorescence spectroscopy has been utilized for the quality analysis of canola and mustard oil along with the effect of heating on their molecular composition has been investigated. Laser diode at 405 nm has been employed directly to oil surface to excite both oil type samples and their emission spectra has been recorded by an in-house developed Fluorosensor. The emission spectra of both oil types unveiled that they contain carotenoids, isomers of vitamin E and chlorophylls that exhibit their fluorescence at 525 and 675/720 nm, and these can be used as markers for their quality assurance. Fluorescence spectroscopy is a fast, reliable and non-destructive analytical technique for the quality assessment of both oil types. Moreover, the effect of temperature on their molecular composition has been investigated by heating them at 110, 120, 130, 140, 150, 170, 180 and 200 °C, each sample for 30 min which was done because both oils are used for cooking and frying. On heating, the deterioration of carotenoids and isomers of vitamin E in both oil types occurred with an increase in the oxidised products. However, it was found that up to 150 °C, both oil types can be used safely for cooking/frying purpose where they do not lose much of their valuable ingredients and up to 180 °C for deep frying, both oils can be used with less deterioration and after that both deteriorated much due to rapid increase of the oxidized products. The portable Fluorosensor, therefore, proved as an excellent device for quality screening of edible oils based on carotenoids and vitamin E.