Pectin demethylation-mediated cell wall Na+ retention positively regulates salt stress tolerance in oilseed rape.

KEY MESSAGE: Integrated phenomics, ionomics, genomics, transcriptomics, and functional analyses present novel insights into the role of pectin demethylation-mediated cell wall Na+ retention in positively regulating salt tolerance in oilseed rape. Genetic variations in salt stress tolerance identified in rapeseed genotypes highlight the complicated regulatory mechanisms. Westar is ubiquitously used as a transgenic receptor cultivar, while ZS11 is widely grown as a high-production and good-quality cultivar. In this study, Westar was found to outperform ZS11 under salt stress. Through cell component isolation, non-invasive micro-test, X-ray energy spectrum analysis, and ionomic profile characterization, pectin demethylation-mediated cell wall Na+ retention was proposed to be a major regulator responsible for differential salt tolerance between Westar and ZS11. Integrated analyses of genome-wide DNA variations, differential expression profiling, and gene co-expression networks identified BnaC9.PME47, encoding a pectin methylesterase, as a positive regulator conferring salt tolerance in rapeseed. BnaC9.PME47, located in two reported QTL regions for salt tolerance, was strongly induced by salt stress and localized on the cell wall. Natural variation of the promoter regions conferred higher expression of BnaC9.PME47 in Westar than in several salt-sensitive rapeseed genotypes. Loss of function of AtPME47 resulted in the hypersensitivity of Arabidopsis plants to salt stress. The integrated multiomics analyses revealed novel insights into pectin demethylation-mediated cell wall Na+ retention in regulating differential salt tolerance in allotetraploid rapeseed genotypes. Furthermore, these analyses have provided key information regarding the rapid dissection of quantitative trait genes responsible for nutrient stress tolerance in plant species with complex genomes.

Effect of Brassica rapa var. rapifera on Insulin Resistance and Inflammatory Analysis of Visceral Adipose Tissue of Obese Wistar Rats Under Glucolipotoxicity Condition / Efecto de Brassica rapa var. rapifera sobre la Resistencia a la Insulina y el Análisis Inflamatorio del Tejido Adiposo Visceral de Ratas Wistar Obesas en Condiciones de Glucolipotoxicidad

SUMMARY: Obesity-related pathophysiologies such as insulin resistance and the metabolic syndrome show a markedly increased risk for type 2 diabetes and atherosclerotic cardiovascular disease. This risk appears to be linked to alterations in adipose tissue function, leading to chronic inflammation and the dysregulation of adipocyte-derived factors. Brassica rapa have been used in traditional medicine for the treatment of several diseases, including diabetes. This study aimed to investigate the effect of nutritional stress induced by a high-fat and high-sucrose diet on the pathophysiology of visceral adipose tissue and the therapeutic effect of Brassica rapa in male Wistar rats. We subjected experimental rats to a high-fat (10 %) high-sucrose (20 %)/per day for 11 months and treated them for 20 days with aqueous extract Br (AEBr) at 200 mg/kg at the end of the experiment. At the time of sacrifice, we monitored plasma and tissue biochemical parameters as well as the morpho-histopathology of visceral adipose tissue. We found AEBr corrected metabolic parameters and inflammatory markers in homogenized visceral adipose tissue and reduced hypertrophy, hyperplasia, and lipid droplets. These results suggest that AEBr enhances anti-diabetic, anti-inflammatory and a protective effect on adipose tissue morphology in type 2 diabetes and obesity.

La fisiopatología relacionadas con la obesidad, como la resistencia a la insulina y el síndrome metabólico, muestran un riesgo notablemente mayor de diabetes tipo 2 y enfermedad cardiovascular aterosclerótica. Este riesgo parece estar relacionado con alteraciones en la función del tejido adiposo, lo que lleva a una inflamación crónica y a la desregulación de los factores derivados de los adipocitos. Brassica rapa se ha utilizado en la medicina tradicional para el tratamiento de varias enfermedades, incluida la diabetes. Este estudio tuvo como objetivo investigar el efecto del estrés nutricional inducido por una dieta rica en grasas y sacarosa sobre la fisiopatología del tejido adiposo visceral y el efecto terapéutico de Brassica rapa en ratas Wistar macho. Sometimos a ratas experimentales a una dieta rica en grasas (10 %) y alta en sacarosa (20 %)/por día durante 11 meses y las tratamos durante 20 días con extracto acuoso de Br (AEBr) a 200 mg/kg al final del experimento. En el momento del sacrificio, monitoreamos los parámetros bioquímicos plasmáticos y tisulares, así como la morfohistopatología del tejido adiposo visceral. Encontramos parámetros metabólicos corregidos por AEBr y marcadores inflamatorios en tejido adiposo visceral homogeneizado y reducción de hipertrofia, hiperplasia y gotitas de lípidos. Estos resultados sugieren que AEBr mejora el efecto antidiabético, antiinflamatorio y protector sobre la morfología del tejido adiposo en la diabetes tipo 2 y la obesidad.

Novel sfericase protease improves amino acid digestibility of soybean meal and rapeseed meal in broiler chickens.

1. Two experiments were conducted to explore the effects of an exogenous sfericase protease on the apparent ileal nutrient digestibility of soybean meal (SBM) and rapeseed meal (RSM) in broiler chickens.2. In each experiment, a total of 256 sixteen-day-old male Cobb 500 broilers were fed one of four semi-purified experimental diets, comprising two different batches (A and B) of samples for either SBM (Exp. 1) or RSM (Exp. 2) without or with an exogenous sfericase (0 or 30,000 NFP/kg). Each experimental diet was fed to eight replicate pens of broiler chickens from 16 to 21 d of age (eight birds per cage), and ileal digesta were collected for measuring the digestibility coefficients.3. In Exp. 1, the amino acid digestibility was greater (P < 0.05) in SBM B compared with SBM A for Arg and Val, and a similar trend (P < 0.1) was observed for Tyr, Leu and Thr. Exogenous sfericase increased (P < 0.10) digestibility of most of amino acids except Gly, His and Trp. There was an interaction between SBM source and sfericase, whereby digestibility of P, N and Asp was increased by sfericase for SBM B but not for SBM A. In Exp. 2, there was no interaction (P > 0.05) between RSM source and sfericase for ileal nutrient digestibility. Digestibility was greater in RSM A compared to RSM B for all non-essential AA and most essential AA (except for Trp), while the reverse was noted for Ca and P (P < 0.05). Exogenous sfericase increased (P < 0.1) digestibility for all amino acids except Cys and Met.4. In conclusion, the current studies showed that both SBM and RSM batches influenced amino acid digestibility. Sfericase protease supplementation increased amino acid digestibility for both SBM and RSM. The digestibility effects were greater in the SBM batch with low digestibility for N and Asp which was in line with an increase in P digestibility.

Evaluation of rapeseed cake as a protein substitute in the feed of edible crickets: A case study using Gryllus assimilis.

This paper aimed to investigate the optimization of Gryllus assimilis farming production by examining the effects of replacing soybean meal with rapeseed cake (25-100%) and supplementing it with rapeseed oil. The results reveal no adverse effects of soybean meal replacement on the feed conversion ratio and weight of the harvested crickets. However, incorporating larger quantities of rapeseed cake into the diet increased crude protein and decreased fat content. Moreover, the composition of fatty acids varied significantly, with increased levels of oleic acid and decreased levels of palmitic acid, while a high rapeseed cake content led to a decrease in the atherogenic and thrombogenic index values. The amino acid composition remained unaffected. In conclusion, the study demonstrates that rapeseed cake can serve as a viable substitute for soybean meal in the diet of Gryllus assimilis.

Targeted mutagenesis of BnTTG1 homologues generated yellow-seeded rapeseed with increased oil content and seed germination under abiotic stress.

Yellow seed is one desirable trait with great potential to improve seed oil quality and yield. The present study surveys the redundant role of BnTTG1 genes in the proanthocyanidins (PA) biosynthesis, oil content and abiotic stress resistance. Stable yellow seed mutants were generated after mutating BnTTG1 by CRISPR/Cas9 genome editing system. Yellow seed phenotype could be obtained only when both functional homologues of BnTTG1 were simultaneously knocked out. Homozygous mutants of BnTTG1 homologues showed decreased thickness and PA accumulation in seed coat. Transcriptome and qRT-PCR analysis indicated that BnTTG1 mutation inhibited the expression of genes involved in phenylpropanoid and flavonoid biosynthetic pathways. Increased seed oil content and alteration of fatty acid (FA) composition were observed in homozygous mutants of BnTTG1 with enriched expression of genes involved in FA biosynthesis pathway. In addition, target mutation of BnTTG1 accelerated seed germination rate under salt and cold stresses. Enhanced seed germination capacity in BnTTG1 mutants was correlated with the change of expression level of ABA responsive genes. Overall, this study elucidated the redundant role of BnTTG1 in regulating seed coat color and established an efficient approach for generating yellow-seeded oilseed rape genetic resources with increase oil content, modified FA composition and resistance to multiple abiotic stresses.

Effects of fermented rapeseed meal on performance, intestinal morphology, the viscosity of intestinal content, phosphorus availability, and egg quality of laying hens.

Fermented rapeseed meal has the potential to partial replace soybean meal in feed mixtures for poultry without a negative impact on the health condition and performance of birds. This is due to the fact that the fermentation process can reduce the amount of antinutritional factors, improve the use of nutrients and impart probiotic properties to rapeseed meal. Therefore, this study was undertaken to investigate the effect of fermented rapeseed meal on the performance, egg quality, intestinal morphometry, the viscosity of intestinal content and total phosphorus availability. A total of 108 Lohmann Brown laying hens at 26 wk of age were used in the 90-day study. All hens were randomly divided into 3 treatment groups, with 12 replicates (cages) each, as follows: control group received no rapeseed meal, the URSM group received 3% unfermented rapeseed meal and the FRSM group received 3% fermented rapeseed meal. In the case of performance, egg traits, sensory evaluation of eggs, the viscosity of intestinal content and the availability of total phosphorus, if the distribution was normal, a 1-way analysis of variance was performed. If the distribution was not normal, the Kruskal-Wallis test was performed. In the case of histomorphometric evaluation of the intestine, if the distribution was normal, the Student t test for independent samples was performed. If not, a Mann-Whitney U test was performed. The performed analyses showed that the supplementation of fermented rapeseed meal had no negative effect on the performance of birds and the quality of eggs. Fermented rapeseed meal was also associated with improved histomorphometric parameters of the small intestine compared to the group receiving unfermented rapeseed meal in the feed. Laying hens from FRSM group were characterized by significantly lower viscosity of intestinal content (P < 0.05) compared to URSM group. Phosphorus in FRSM group was significantly more available to the birds (P < 0.05) compared to URSM group. These results suggest that supplementation with fermented rapeseed meal may be beneficial, especially in times of unstable prices of soybean meal and problems with its availability.

Genomics of predictive radiation mutagenesis in oilseed rape: modifying seed oil composition.

Rapeseed is a crop of global importance but there is a need to broaden the genetic diversity available to address breeding objectives. Radiation mutagenesis, supported by genomics, has the potential to supersede genome editing for both gene knockout and copy number increase, but detailed knowledge of the molecular outcomes of radiation treatment is lacking. To address this, we produced a genome re-sequenced panel of 1133 M2 generation rapeseed plants and analysed large-scale deletions, single nucleotide variants and small insertion-deletion variants affecting gene open reading frames. We show that high radiation doses (2000 Gy) are tolerated, gamma radiation and fast neutron radiation have similar impacts and that segments deleted from the genomes of some plants are inherited as additional copies by their siblings, enabling gene dosage decrease. Of relevance for species with larger genomes, we showed that these large-scale impacts can also be detected using transcriptome re-sequencing. To test the utility of the approach for predictive alteration of oil fatty acid composition, we produced lines with both decreased and increased copy numbers of Bna.FAE1 and confirmed the anticipated impacts on erucic acid content. We detected and tested a 21-base deletion expected to abolish function of Bna.FAD2.A5, for which we confirmed the predicted reduction in seed oil polyunsaturated fatty acid content. Our improved understanding of the molecular effects of radiation mutagenesis will underpin genomics-led approaches to more efficient introduction of novel genetic variation into the breeding of this crop and provides an exemplar for the predictive improvement of other crops.

Pectin polysaccharide contribution to oleosome extraction after wet milling of rapeseed.

Oleosomes are lipid composites providing energy storage in oilseeds. They possess a unique structure, comprised of a triglyceride core stabilized by a phospholipid-protein membrane, and they have shown potential to be used as ingredients in several food applications. Intact oleosomes are extracted by an aqueous process which includes soaking, milling, and gravitational separation. However, the details of the complexes formed between oleosomes, proteins and pectin polysaccharides during this extraction are not known. It was hypothesized that pectins play an important role during the oleosome separation, and different proteins will be complexed on the surface of the oleosomes, depending on the pH of extraction. Rapeseed extracts were treated with and without pectinase (Pectinex Ultra SP-L) and extracted at pH 5.7 or 8.5, as this will affect electrostatic complexation. Acidic conditions led to co-extraction of storage proteins, structured as dense oleosome emulsions, stabilized by a network of proteins and polysaccharides. Pectinase intensified this effect, highlighting pectic polysaccharides' role in bridging interactions among proteins and oleosomes under acidic conditions. The presence of this dense interstitial layer around the oleosomes protected them from coalescence during extraction. Conversely, under alkaline conditions, the extraction process yielded more purified oleosomes characterized by a larger particle size, most likely due to coalescence. Nevertheless, pectinase addition at pH 8.5 mitigated coalescence tendencies. These results contribute to a better understanding of the details of the colloidal complexes formed during extraction and can be used to modulate the composition of the extracted fractions, with significant consequences not only for yields and purity but also for the functional properties of the ingredients produced.

Selenium-oxidizing Agrobacterium sp. T3F4 decreases arsenic uptake by Brassica rapa L. under a native polluted soil.

Toxic arsenic (As) and trace element selenium (Se) are transformed by microorganisms but their complex interactions in soil-plant systems have not been fully understood. An As- and Se- oxidizing bacterium, Agrobacterium sp. T3F4, was applied to a native seleniferous As-polluted soil to investigate As/Se uptake by the vegetable Brassica rapa L. and As-Se interaction as mediated by strain T3F4. The Se content in the aboveground plants was significantly enhanced by 34.1%, but the As content was significantly decreased by 20.5% in the T3F4-inoculated pot culture compared to the control (P < 0.05). Similar result was shown in treatment with additional 5 mg/kg of Se(IV) in soil. In addition, the As contents in roots were significantly decreased by more than 35% under T3F4 or Se(IV) treatments (P<0.05). Analysis of As-Se-bacterium interaction in a soil simulation experiment showed that the bioavailability of Se significantly increased and As was immobilized with the addition of the T3F4 strain (P < 0.05). Furthermore, an As/Se co-exposure hydroponic experiment demonstrated that As uptake and accumulation in plants was reduced by increasing Se(IV) concentrations. The 50% growth inhibition concentration (IC50) values for As in plants were increased about one-fold and two-fold under co-exposure with 5 and 10 µmol/L Se(IV), respectively. In conclusion, strain T3F4 improves Se uptake but decreases As uptake by plants via oxidation of As and Se, resulting in decrease of soil As bioavailability and As/Se competitive absorption by plants. This provides a potential bioremediation strategy for Se biofortification and As immobilization in As-polluted soil.

Allelopathic effects of Eucalyptus extract and wood vinegar on germination and sprouting of rapeseed (Brassica rapa L.).

Eucalyptus plantations are usually characterized by low biodiversity due to allelopathy effects. Wood vinegar is considered a complex growth regulator that can promote plant growth at low concentrations. However, there is information scarcity about the co-application of eucalypt leaf water extract and wood vinegar on plants. This study aimed at clarifying whether wood vinegar can protect seed germination against suppression by eucalypt-induced allelopathy. We examined germination behavior and seedling elongation characteristics in rapeseed (Brassica rapa L.) treated with different solutions of wood vinegar and eucalypt leaf water extract. The results showed that eucalypt leaf water extracts, wood vinegar solutions, and their mixture allelopathically suppressed seed germination rate. After rapeseed sprouting, eucalypt leaf water extracts promoted root elongation, stem elongation, and fresh weight elongation. Malondialdehyde content was also lower under the influence of eucalypt leaf water extract. Mixture of high concentration of eucalypt leaf water extract and lower concentration of wood vinegar significantly promoted root elongation. Therefore, both eucalypt leaf water extract and wood vinegar are complex plant growth regulators, which can be used to inhibit or stimulate plants at different ontogenic stages. During the seed germination period, both eucalypt leaf extracts and wood vinegar could be used as weed inhibitors. Conversely, during the period of sprouting (seedling establishment), low concentrations of eucalypt leaf extracts and wood vinegar can promote growth.

Transcriptomic Analysis of the Reduction in Seed Oil Content through Increased Nitrogen Application Rate in Rapeseed (Brassica napus L.).

Nitrogen is essential for improving the seed oil yield of rapeseed (Brassica napus L.). However, the molecular mechanism by which increased nitrogen rates impact seed oil content is largely unknown. Therefore, a field experiment was conducted to determine how three nitrogen application rates (120, 240, and 360 kg ha-1) regulated seed oil content via transcriptomic analysis. The results showed that the seed yield and the protein and total N contents increased from N1 to N3, with average increases of 57.2%, 16.9%, and 79.5%, respectively. However, the seed oil content significantly decreased from N1 to N3, with an average decrease of 8.6%. These results were repeated over a number of years. The quantity of oil protein bodies observed under a transmission electron microscope was in accordance with the ultimate seed oil and protein contents. As the nitrogen application rate increased, a substantial number of genes involved in the photosynthesis, glycolysis, and phenylpropanoid biosynthesis pathways were up-regulated, as were TF families, such as AP2/ERF, MYB, and NAC. The newly identified genes were mainly involved in carbohydrate, lipid, and amino acid metabolism. Metabolic flux analysis showed that most of the genes involved in glycolysis and fatty acid biosynthesis had higher transcript levels in the early development stages. Our results provide new insights into the molecular regulation of rapeseed seed oil content through increased nitrogen application rates.

Research Progress on the Effect of Nitrogen on Rapeseed between Seed Yield and Oil Content and Its Regulation Mechanism.

Rapeseed (Brassica napus L.) is one of the most important oil crops in China. Improving the oil production of rapeseed is an important way to ensure the safety of edible oil in China. Oil production is an important index that reflects the quality of rapeseed and is determined by the oil content and yield. Applying nitrogen is an important way to ensure a strong and stable yield. However, the seed oil content has been shown to be reduced in most rapeseed varieties after nitrogen application. Thus, it is critical to screen elite germplasm resources with stable or improved oil content under high levels of nitrogen, and to investigate the molecular mechanisms of the regulation by nitrogen of oil accumulation. However, few studies on these aspects have been published. In this review, we analyze the effect of nitrogen on the growth and development of rapeseed, including photosynthetic assimilation, substance distribution, and the synthesis of lipids and proteins. In this process, the expression levels of genes related to nitrogen absorption, assimilation, and transport changed after nitrogen application, which enhanced the ability of carbon and nitrogen assimilation and increased biomass, thus leading to a higher yield. After a crop enters the reproductive growth phase, photosynthates in the body are transported to the developing seed for protein and lipid synthesis. However, protein synthesis precedes lipid synthesis, and a large number of photosynthates are consumed during protein synthesis, which weakens lipid synthesis. Moreover, we suggest several research directions, especially for exploring genes involved in lipid and protein accumulation under nitrogen regulation. In this study, we summarize the effects of nitrogen at both the physiological and molecular levels, aiming to reveal the mechanisms of nitrogen regulation in oil accumulation and, thereby, provide a theoretical basis for breeding varieties with a high oil content.

Enzymatic fermentation of rapeseed cake significantly improved the soil environment of tea rhizosphere.

BACKGROUND: Rapeseed cake is an important agricultural waste. After enzymatic fermentation, rapeseed cake not only has specific microbial diversity but also contains a lot of fatty acids, organic acids, amino acids and their derivatives, which has potential value as a high-quality organic fertilizer. However, the effects of fermented rapeseed cake on tea rhizosphere microorganisms and soil metabolites have not been reported. In this study, we aimed to elucidate the effect of enzymatic rapeseed cake fertilizer on the soil of tea tree, and to reveal the correlation between rhizosphere soil microorganisms and nutrients/metabolites. RESULTS: The results showed that: (1) The application of enzymatic rapeseed cake increased the contents of soil organic matter (OM), total nitrogen (TN), total phosphorus (TP), available nitrogen (AN), and available phosphorus (AP); increased the activities of soil urease (S-UE), soil catalase (S-CAT), soil acid phosphatase (S-ACP) and soil sucrase (S-SC); (2) The application of enzymatic rapeseed cake increased the relative abundance of beneficial rhizosphere microorganisms such as Chaetomium, Inocybe, Pseudoxanthomonas, Pseudomonas, Sphingomonas, and Stenotrophomonas; (3) The application of enzymatic rapeseed cake increased the contents of sugar, organic acid, and fatty acid in soil, and the key metabolic pathways were concentrated in sugar and fatty acid metabolisms; (4) The application of enzymatic rapeseed cake promoted the metabolism of sugar, organic acid, and fatty acid in soil by key rhizosphere microorganisms; enzymes and microorganisms jointly regulated the metabolic pathways of sugar and fatty acids in soil. CONCLUSIONS: Enzymatic rapeseed cake fertilizer improved the nutrient status and microbial structure of tea rhizosphere soil, which was beneficial for enhancing soil productivity in tea plantations. These findings provide new insights into the use of enzymatic rapeseed cake as an efficient organic fertilizer and expand its potential for application in tea plantations.

NPs-Ca promotes Cd accumulation and enhances Cd tolerance of rapeseed shoots by affecting Cd transfer and Cd fixation in pectin.

The use of rapeseed (Brassica napus) as a hyperaccumulator plant has shown great promise for the remediation of cadmium (Cd) contaminated soils. Nanosized materials (NPs) have been shown to mitigate heavy metal toxicity in plants, but it is unknown how l-aspartate nano-calcium (NPs-Ca) affects Cd uptake, transport, and tolerance in rapeseed. A soil pot experiment was conducted with two treatments: a control treatment (CK) with 2.16 g CaCl2 and NPs-Ca treatment with 6.00 g NPs-Ca, to evaluate the effects and mechanisms of NPs-Ca on Cd tolerance in rapeseed. Compared to CaCl2, NPs-Ca promoted Cd transportation from roots to shoots by up-regulating the expression of Cd transport genes (ABCC12, HMA8, NRAM6, ZIP6, CAX4, PCR2, and HIP6). Therefore, NPs-Ca increased Cd accumulation in rapeseed shoots by 39.4%. Interestingly, NPs-Ca also enhanced Cd tolerance in the shoots, resulting in lower hydrogen peroxide (H2O2) accumulation and proline content, as well as higher antioxidant enzyme activities (POD, CAT). Moreover, NPs-Ca reduced the activity of pectin-degrading enzymes (polygalacturonase: PG, ß-galactosidase: ß-GAL), promoted the activity of pectin methyl esterase (PME), and changed transcription levels of related genes (PME, PMEI, PG, PGIP, and ß-GAL). NPs-Ca treatment also significantly increased the Cd content in cell walls by 59.8%, that is, more Cd was immobilized in cell walls, and less Cd entered organelles in shoots of NPs-Ca treatment due to increased pectin content and degree of pectin demethylation. Overall, NPs-Ca increased Cd accumulation in rapeseed shoots by promoting Cd transport from roots to shoots. And meantime, NPs-Ca enhanced Cd tolerance of shoots by inhibiting pectin degradation, promoting pectin demethylation and increasing Cd fixation in pectin. These findings suggest that NPs-Ca can improve the potential of rapeseed as a hyperaccumulator for the remediation of Cd-contaminated soil and the protection of the environment. Furthermore, the study provides a theoretical basis for the application of NPs-Ca in the phytoremediation of Cd-contaminated soils with hyperaccumulating plants.

Multiomics analysis reveals a link between Brassica-specific miR1885 and rapeseed tolerance to low temperature.

Brassica crops include various edible vegetable and plant oil crops, and their production is limited by low temperature beyond their tolerant capability. The key regulators of low-temperature resistance in Brassica remain largely unexplored. To identify posttranscriptional regulators of plant response to low temperature, we performed small RNA profiling, and found that 16 known miRNAs responded to cold treatment in Brassica rapa. The cold response of seven of those miRNAs were further confirmed by qRT-PCR and/or northern blot analyses. In parallel, a genome-wide association study of 220 accessions of Brassica napus identified four candidate MIRNA genes, all of which were cold-responsive, at the loci associated with low-temperature resistance. Specifically, these large-scale data analyses revealed a link between miR1885 and the plant response to low temperature in both B. rapa and B. napus. Using 5' rapid amplification of cDNA ends approach, we validated that miR1885 can cleave its putative target gene transcripts, Bn.TIR.A09 and Bn.TNL.A03, in B. napus. Furthermore, overexpression of miR1885 in Semiwinter type B. napus decreased the mRNA abundance of Bn.TIR.A09 and Bn.TNL.A03 and resulted in increased sensitivity to low temperature. Knocking down of miR1885 in Spring type B. napus led to increased mRNA abundance of its targets and improved rapeseed tolerance to low temperature. Together, our results suggested that the loci of miR1885 and its targets could be potential candidates for the molecular breeding of low temperature-tolerant Spring type Brassica crops.

A novel type of Brassica napus with higher stearic acid in seeds developed through genome editing of BnaSAD2 family.

KEY MESSAGE: Modifications of multiple copies of the BnaSAD2 gene family with genomic editing technology result in higher stearic acid content in the seed of polyploidy rapeseed. Solid fats from vegetable oils are widely used in food processing industry. Accumulating data showed that stearic acid is more favorite as the major composite among the saturate fatty acids in solid fats in considerations of its effects on human health. Rapeseed is the third largest oil crop worldwide, and has potential to be manipulated to produce higher saturated fatty acids as raw materials of solid fats. Toward that end, we identified four SAD2 gene family members in B. napus genome and established spatiotemporal expression pattern of the BnaSAD2 members. Genomic editing technology was applied to mutate all the copies of BnaSAD2 in this allopolyploid species and mutants at multiple alleles were generated and characterized to understand the effect of each BnaSAD2 member on blocking desaturation of stearic acid. Mutations occurred at BnaSAD2.A3 resulted in more dramatic changes of fatty acid profile than ones on BnaSAD2.C3, BnaSAD2.A5 and BnaSAD2.C4. The content of stearic acid in mutant seeds with single locus increased dramatically with a range of 3.1-8.2%. Furthermore, combination of different mutated alleles of BnaSAD2 resulted in more dramatic changes in fatty acid profiles and the double mutant at BnaSAD2.A3 and BnaSAD2.C3 showed the most dramatic phenotypic changes compared with its single mutants and other double mutants, leading to 11.1% of stearic acid in the seeds. Our results demonstrated that the members of BnaSAD2 have differentiated in their efficacy as a Δ9-Stearoyl-ACP-Desaturase and provided valuable rapeseed germplasm for breeding high stearic rapeseed oil.

Performance and antioxidant traits of broiler chickens fed with diets containing rapeseed or flaxseed oil and optimized quercetin.

This study evaluated the effect of quercetin (Q) added to feed mixtures, at concentrations directly optimized for the peroxidability of dietary rapeseed (RO) and flaxseed oil (FLO), on performance and selected biomarkers of oxidative stress of broiler chickens. Ninety-six one-day-old Ross 308 broiler chicken males were randomly assigned to four groups (six replicates per treatment, four birds per cage, n = 24 per group): Group RO received diets containing rapeseed oil (RO) and group FLO received diets containing flaxseed oil (FLO); Group RO_Q and group FLO_Q received these same diets containing RO or FLO oils, supplemented with optimized quercetin (Q). Blood, pectoral muscles, and liver samples of chickens were collected after 35 days to determine: (1) the global indicators of antioxidant capacity: ferric reducing antioxidant power (FRAP), antiradical activity (DPPH·/ABTS·+), total antioxidant status (TAS), and glutathione peroxidase (GSH-Px); (2) the activity of the antioxidant enzymes catalase (CAT) and superoxide dismutase (SOD); and (3) the concentration of malondialdehyde (MDA). Data showed that the FLO diet did not affect the final performance parameters in relation to RO, but the optimized Q tended to improve the total body weight gain and the final body weight of broiler chickens (P = 0.10). The antioxidant traces analyzed in the blood (GSH-Px), plasma (FRAP, ABTS·+, DPPH·, TAS), serum (DPPH·), and pectoral muscles (SOD, CAT) of chickens were not altered by either Oil or Q factor. FLO supplementation increased MDA content in the liver of chickens (P < 0.05) and increased liver CAT activity, which was not improved by optimized Q. Meanwhile, the Oil × Q interaction suggests that optimized Q could reduce the liver burden and negative effects of oxidized lipid by-products associated with FLO diets. Our results indicate that optimizing the addition of natural polyphenols to feed may be a valuable alternative to the application of polyphenolic antioxidants in animal nutrition, allowing for an economical use of the antioxidant additives when customized to the peroxidability of fat sources, which is line to the conception of sustainable development covering 'The European Green Deal' and 'Farm to Fork Strategy'.

Rapid and Low-Cost Quantification of Adulteration Content in Camellia Oil Utilizing UV-Vis-NIR Spectroscopy Combined with Feature Selection Methods.

This study aims to explore the potential use of low-cost ultraviolet-visible-near infrared (UV-Vis-NIR) spectroscopy to quantify adulteration content of soybean, rapeseed, corn and peanut oils in Camellia oil. To attain this aim, test oil samples were firstly prepared with different adulterant ratios ranging from 1% to 90% at varying intervals, and their spectra were collected by an in-house built experimental platform. Next, the spectra were preprocessed using Savitzky-Golay (SG)-Continuous Wavelet Transform (CWT) and the feature wavelengths were extracted using four different algorithms. Finally, Support Vector Regression (SVR) and Random Forest (RF) models were developed to rapidly predict adulteration content. The results indicated that SG-CWT with decomposition scale of 25 and the Iterative Variable Subset Optimization (IVSO) algorithm can effectively improve the accuracy of the models. Furthermore, the SVR model performed best for predicting adulteration of camellia oil with soybean oil, while the RF models were optimal for camellia oil adulterated with rapeseed, corn, or peanut oil. Additionally, we verified the models' robustness by examining the correlation between the absorbance and adulteration content at certain feature wavelengths screened by IVSO. This study demonstrates the feasibility of using low-cost UV-Vis-NIR spectroscopy for the authentication of Camellia oil.

Effects of faba bean, blue lupin and rapeseed meal supplementation on amino acid metabolism of dairy cows fed grass silage-based diets.

Information about the amino acid (AA) supply of locally produced protein supplements to dairy cow metabolism is needed to design sustainable diets for milk production. In this dairy cow experiment, grass silage and cereal-based diets supplemented with isonitrogenous amounts of rapeseed meal (RSM), faba beans (FB) and blue lupin seeds (BL) were compared with a control diet (CON) without protein supplementation. The diets were arranged as a 4 × 4 Latin Square using periods of 21 days, and four rumen-cannulated Nordic Red dairy cows were used in the experiment. The intake of all AAs increased in response to protein supplementation and was for many individual AAs higher when RSM rather than the grain legumes FB and BL were fed. The total AA flow at the omasal canal was 3 026, 3 371, 3 373 and 3 045 g/day for cows fed CON, RSM, FB and BL, respectively, but only RSM resulted in higher milk protein output. This may be explained by the higher provision of essential AA for milk protein synthesis when RSM was fed. The cows fed FB showed some positive features such as a tendency for greater omasal flow of branched-chain AA compared with BL. Overall, low plasma methionine and/or glucose concentrations in all treatments suggest that their supply was possibly limiting further production responses under the dietary conditions of the current study. It seems that the benefits of grain legume supplementation are limited when high-quality grass silage and cereal-based diets are used as the basal diet, but higher responses in amino acid supply and subsequent production responses can be expected when RSM is used.