Genetic Diversity of Testa Pigments and RAPD Marker of Yellow-Seeded Rapeseed (Brassica napus L.)

14 yellow-seeded rapeseed lines (Brassia napus L.) from different genetic sources were used to analyze diversity of testa pigments content, oil and protein content, and RAPD markers. The results showed that the anthocyanin and melanin were the most important pigments in testa and their content were responsible for the variation in seed color ranging from orange to black yellow, 14 yellow-seeded lines could be classified into 3 groups: high anthocyanin content group with anthocyanin content over 2. 54 mg g-1 DW, the seed color was light yellow or orange; low pigments content group with low content of anthocyanin and melanin, the testa was transparent and the seed color was light yellow, greenish yellow or twany; high melanin content group with melanin content over 178. 4U(A290nm) , the testa was black, the seed color was black yellow. Oil content changed from 36.2% to 45. 5%, protein content from 21.1% to 27.7% , and the correlation analysis revealed that the oil content is highly significantly negatively correlated with the protein content. The cluster analysis showed that the extensive genetic variation existed among 14 yellow-seeded lines by using unweighted paired group method with arithmetic average (UPGMA) based on RAPD markers which were amplified with decamer primers, the genetic similarity among them ranged from 0. 25 to 0.909, and 14 yellow-seeded lines could put into 2 clusters corresponding to genome difference.

Identification and virulence test of a new pathogen that causes verticillium striping on rapeseed in northwestern China

Five stems of rapeseed with abundant black microsclerotia were collected from Huangyuan County of Qinghai Province,China,and fungal isolates were obtained from the stems.They were identified based on morphology,molecular features and specific PCR detection.The results showed that the 10 fungal isolates belonged to Verticillium longisporum lineage A1/D3.One of the 10 isolates(HW7-1)was tested for virulence on three species of rapeseed,including B.napus Zhongshuang 9,B.rapa Qingyou 9 and B.juncea Tayou 2 by conidia inoculation of HW7-1 on roots of young seedlings.Control seedlings were inoculated with V.dahliae conidia or water alone.The seedlings of these treatments were transplanted in culture mix and incubated in a growth chamber(20℃).Results suggested that the control seedlings of three cultivars appeared quite healthy,while the seedlings inoculated with HW7-1 turned yellowing leaves,seedling stunting or even death after 22 days post-inoculation.V.longisporum was re-isolated from he yellow leaves,thus fulfilling Koch's postulates.Moreover,compared to the control treatments,inoculation with HW7-1 caused flowering delay and seed yield reduction on Tayou 2 with production of microsclerotia on the stems.To our knowledge,this is the first report of V.longisporum lineage A1/D3 on rapeseed in northwestern China.

Utilizing auxin dwarf genes to optimize seed yield and lodging resistance in rapeseed

Direct-seeding rapeseed production at high plant density raises the risk of lodging.We investigated the use of dwarf genes to improve rapeseed plant architecture to balance yield and lodging.Three genotypes with different plant architectures(dwarf sca^(HS5),semi-dwarf+/sca^(HS5),and tall ^(HS5))were evaluated under varying nitrogen rates(N1,N2,and N3:120,240,and 360 kg N ha^(-1))and plant densities(D1,D2,and D3:15,45,and 75 plants m^(-2))from 2019 to 2022.The results showed that increasing N rate positively influenced yield while decreasing lodging resistance in all genotypes.Increasing plant density(D2-D3)enhanced lodging resistance and yield in sca^(HS5) and+/sca^(HS5),but reduced yield in ^(HS5).Compared to the two parents,+/sca^(HS5) exhibited moderate expressions of IAA3,GH3.15,and SAUR30 in stems under N2D3,resulting in reduced plant height and increased compactness.Additionally,+/sca^(HS5) had a thicker silique layer than ^(HS5) by 14.7%,and it had a significant correlation between branch height/angle and yield.Increasing N rate led to increased lignin and pectin contents,while cellulose content decreased.Increasing plant density resulted in greater stem cellulose content and CSLA3/7 expression in sca^(HS5) and+/sca^(HS5),but decreased in ^(HS5).Compared to ^(HS5),+/sca^(HS5) exhibited higher expressions of ARAD1 and GAUT4,along with a 51.1%increase in pectin content,leading to improved lodging resistance under N2D3.Consequently,+/sca^(HS5) showed a 46.4%higher yield and 38.9%lodging resistance than ^(HS5) under N2D3,while sca^(HS5) demonstrated strong lodging resistance but lower yield potential.Overall,this study underscores the potential of utilizing auxin dwarf genes to optimize the trade-off between yield and lodging resistance in rapeseed and the possibility of maximizing yield potential by optimizing the plant architecture of+/sca^(HS5) through nitrogen reduction and dense planting.

Screening of glucosinolates degrading lactic acid bacteria and their utilization in rapeseed meal fermentation

Rapeseed meal is a promising food ingredient, but its utilization is limited by the presence of some potentially harmful ingredients, such as glucosinolates. Fermentation is a cost-effective method of detoxication but a food-grade starter culture with glucosinolates degradation capacity is required. In this study, 46 strains of lactic acid bacteria from traditional paocai brines were screened for their ability to glucosinolate degradation. The results showed that more than 50% of the strains significantly degraded glucosinolates. Two strains of Lactiplantibacillus(p7 and s7) with high capacity of glucosinolates degradation through producing enzymes were identified. Then,an optimized condition for rapeseed meal fermentation by p7 was established to degrade glucosinolates, which can achieve about 80% degradation. UPLC/Q-TOF-MS analysis showed that the degradation rate of individual glucosinolates was different and the degradation rate of gluconapin and progoitrin in rapeseed meal can reach more than 90%. Meanwhile, fermentation with p7 can improve safety of rapeseed meal by inhibiting the growth of Enterobacteriaceae and improve its nutritional properties by degrading phytic acid. The in vitro digestion experiments showed that the content of glucosinolates in rapeseed meal decreased significantly during gastric digestion. Meanwhile, fermentation with p7 can greatly improve the release of soluble protein and increase the contents of free essential amino acids, such as lysine(increased by 12 folds) and methionine(increased by 10 folds).

Comparative assessment of the frying efficiency of standard and low linolenic rapeseed oils: Principal Component Analysis (PCA)

In this research,the performance of regular rapeseed oil(RSO)and modified low-linolenic rapeseed oil(LLRO)during frying was assessed using a frying procedure that commonly found in fast-food restaurants.Key physicochemical attributes of these oils were investigated.RSO and LLRO differed for initial linolenic acid(12.21%vs.2.59%),linoleic acid(19.15%vs.24.73%).After 6 successive days frying period of French fries,the ratio of linoleic acid to palmitic acid dropped by 54.49%in RSO,higher than that in LLRO(51.54%).The increment in total oxidation value for LLRO(40.46 unit)was observed to be significantly lower than those of RSO(42.58 unit).The changes in carbonyl group value and iodine value throughout the frying trial were also lower in LLRO compared to RSO.The formation rate in total polar compounds for LLRO was 1.08%per frying day,lower than that of RSO(1.31%).In addition,the formation in color component and degradation in tocopherols were proportional to the frying time for two frying oils.Besides,a longer induction period was also observed in LLRO(8.87 h)compared to RSO(7.68 h)after frying period.Overall,LLRO exhibited the better frying stability,which was confirmed by principal component analysis(PCA).

Physiological Differences Between Yellow-Seeded and Black-Seeded Rapeseeds (Brassica napus L.) with Different Testa Characteristics During Artificial Ageing

Yellow-seeded rapeseed (Brassica napus L.) is a new kind of breeding resources with yellow color, increased oil and protein content and less unwanted crude fiber content due to the thinner and transparent testa compared with traditional black or brown-seeded rapeseed. To analyze the longevity of the yellow-seeded rapeseed during storage, the physiological differences between the yellow and black-seeded near-isogenic lines were studied by artificial ageing method. The testa rate, anthocyanin content and melanin content of yellow-seeded rapeseeds decreased by 20.1, 25.2, and 80.4% respectively than black-seeded rapeseed. During artificial ageing, the yellow-seeded rapeseed showed significantly different effect of ageing compared with the black-seeded, as demonstrated by faster deterioration with lower germination percentage, seed vigour index, reducing sugar and soluble protein contents than the black-seeded, as well as a drastic increase in electrical conductivity, malnodialdehyde (MDA) content and a rapid decrease in activities of antioxidant enzymes catalase (CAT) and superoxide dismutase (SOD). The results suggested that the transparent testa of the yellow-seeded rapeseed lost some abilities to protect the embryo against adverse environmental conditions and thus led to a poor storability.

Influence of two-stage harvesting on the properties of cold-pressed rapeseed(Brassica napus L.)oils

Rapeseed(Brassica napus L.)harvesting method is critical since it significantly determines the seed yield,oil quality,and industrial efficiency.This study investigated the influences of harvesting methods on the quality of cold-pressed rapeseed oil of two varieties.Oil color,peroxide value(POV),tocopherol content,fatty acid composition,and polarity of total polyphenols(PTP)contents of two rapeseed varieties in Huanggang and Xiangyang were compared through artificially simulated combined harvesting and two-stage harvesting.Results showed significant differences in the quality of rapeseed oil between the two harvesting methods.The red value(R-value),POV,total tocopherol contents,linoleic and linolenic acid content,and PTP content of the pressed rapeseed oil prepared by the combined harvesting method were about 27.6,5.7,15.8,2.0,0.5,and 28.6%lower than those of the oil produced from the two-stage harvesting method,respectively.Xiangyang and Huayouza62 performed better in the two regions and two varieties,respectively.To sum up,the rapeseed oil obtained 41–44 days after final flowering of combined harvesting,35 days after final flowering,and six days of post-ripening of the two-stage harvesting had the best quality.

Development of a target capture sequencing SNP genotyping platform for genetic analysis and genomic breeding in rapeseed

Rapeseed(Brassica napus)is an oil crop grown worldwide,making it a key plant species in molecular breeding research.However,the complexity of its polyploid genome increases sequencing costs and reduces sequencing accuracy.Target capture coupled with high-throughput sequencing is an efficient approach for detecting genetic variation at genomic regions or loci of interest.In this study,588 resequenced accessions of rapeseed were used to develop a target capture sequencing SNP genotyping platform named BnaPan50T.The platform comprised 54,765,with 54,058 resequenced markers from the pan-genome,and 855 variant trait-associated markers for 12 agronomic traits.The capture quality of BnaPan50T was demonstrated well in 12 typical accessions.Compared with a conventional genotyping array,BnaPan50T has a high SNP density and a high proportion of SNPs in unique physical positions and in annotated functional genes,promising wide application.Target capture sequencing and wholegenome resequencing in 90 doubled-haploid lines yielded 60%specificity,78%uniformity within tenfold coverage range,and 93%genotyping accuracy for the platform.BnaPan50T was used to construct a genetic map for quantitative trait loci(QTL)mapping,identify 21 unique QTL,and predict several candidate genes for yield-related traits in multiple environments.A set of 132 core SNP loci was selected from BnaPan50T to construct DNA fingerprints and germplasm identification resources.This study provides genomics resources to support target capture sequencing,genetic analysis and genomic breeding of rapeseed.

National Protection Zone for Rapeseed Production and Industrial Cluster of Rape in Hubei Province

Hubei Province is the main winter rapeseed producing area in China,and has the national advantage and characteristic industrial cluster of rape.The protection zone for the production of important agricultural products of rapeseed in Hubei Province also ranks first as that in Hunan Province.This paper studied the winter rape industry,the important agricultural product production protection zone of rapeseed,the provincial double-low high-quality rape protection zone and the industrial cluster of rape in Jianghan Plain in Hubei Province,and analyzed the main problems existing in the development of rapeseed production protection zone and industrial cluster of rape.Finally,it came up with strategies including promoting national rape varieties,overcoming the problem of rape"agricultural chip",promoting the"rice+rape"rotation model,and strengthening the protection of intellectual property rights of brand marks.

Mapping winter rapeseed in South China using Sentinel-2 data based on a novel separability index

Large-scale crop mapping using remote sensing data is of great significance for agricultural production, food security and the sustainable development of human societies. Winter rapeseed is an important oil crop in China that is mainly distributed in the Yangtze River Valley. Traditional winter rapeseed mapping practices are insufficient since they only use the spectral characteristics during the critical phenological period of winter rapeseed, which are usually limited to a small region and cannot meet the needs of large-scale applications. In this study, a novel phenology-based winter rapeseed index(PWRI) was proposed to map winter rapeseed in the Yangtze River Valley. PWRI expands the date window for distinguishing winter rapeseed and winter wheat, and it has good separability throughout the flowering period of winter rapeseed. PWRI also improves the separability of winter rapeseed and winter wheat, which traditionally have been two easily confused winter crops. A PWRI-based method was applied to the Middle Reaches of the Yangtze River Valley to map winter rapeseed on the Google Earth Engine platform. Time series composited Sentinel-2 data were used to map winter rapeseed with 10 m resolution. The mapping achieved a good result with overall accuracy and kappa coefficients exceeding 92% and 0.85, respectively. The PWRI-based method provides a new solution for high spatial resolution winter rapeseed mapping at a large scale.

Quick and Accurate Counting of Rapeseed Seedling with Improved YOLOv5s and Deep-Sort Method

The statistics of the number of rapeseed seedlings are very important for breeders and planters to conduct seed quality testing,field crop management and yield estimation.Calculating the number of seedlings is inefficient and cumbersome in the traditional method.In this study,a method was proposed for efficient detection and calculation of rapeseed seedling number based on improved you only look once version 5(YOLOv5)to identify objects and deep-sort to perform object tracking for rapeseed seedling video.Coordinated attention(CA)mechanism was added to the trunk of the improved YOLOv5s,which made the model more effective in identifying shaded,dense and small rapeseed seedlings.Also,the use of the GSConv module replaced the standard convolution at the neck,reduced model parameters and enabled it better able to be equipped for mobile devices.The accuracy and recall rate of using improved YOLOv5s on the test set by 1.9%and 3.7%compared to 96.2%and 93.7%of YOLOv5s,respectively.The experimental results showed that the average error of monitoring the number of seedlings by unmanned aerial vehicles(UAV)video of rapeseed seedlings based on improved YOLOv5s combined with depth-sort method was 4.3%.The presented approach can realize rapid statistics of the number of rapeseed seedlings in the field based on UAV remote sensing,provide a reference for variety selection and precise management of rapeseed.

Growth Dynamics of Rapeseed Leaves Based on MATLAB

[Ojective]The study aimed to simulate the growth dynamics of rapeseed leaves.[Method]Using nine rapeseed cultivars as experimental materials,the accumulation situation of rapeseed leaves was recorded and their growth dynamics were simulated by using MATLAB program.[Result]The leaf emergence rates of each genotype materials at different growth stages were determined to be:after spring>seedling stage>overwintering stage.All the related indices of the Logistic equation were higher than 0.976 7,commendably reflecting the growth dynamics of rapeseed leaves.[Conclusion]The study provided theoretical basis for the intelligentized cultivation and management and showed the practical guiding significance for the production of high quality rapeseed.

Analysis of Yield and Disease Resistance Traits of New Winter Rapeseed Varieties over the Past Twenty Years in China

Based on the results of the national regional trail for winter rapeseed in four groups,including the upper reaches,middle reaches and lower reaches of Yangtze River and Huang-Huai region in the past twenty years,new varieties of winter rapeseed showed upward trend in average yield,the yield level in Huang-huai group was higher than other groups.The changes of average effective pods per plant were not significant in any group,but the number of grain per pod and 1 000-grain weight showed increase trend.However,the increment of grains per pod in three Yangtze River groups was higher than that in Huang-Huai River group,while the increment of 1 000-grain weight was just opposite.The incidence rate of Sclerotinia sclerotiorum and viral disease in new varieties of winter rapeseed obviously decreased,and the incidence rate of S.sclerotiorum decreased lower than 5%.Genetic improvement for winter rapeseed should be focus on the number of grain per pod and 1 000-grain weight in the future,and yield level of new varieties in all four groups is expected to increase.

Study on the Optimization of Two-dimensional Electrophoresis Technology System for Rapeseed Proteome

[Objective] The research aimed to establish the two-dimensional electrophoresis（2-DE）technology which was suitable for the rapeseed proteome research.[Method] Xiangyou 17 was as the material.The sample preparation method,gel concentration and loading amount,etc.in 2-DE technology were optimized.[Result] The best extraction method of total protein of rapeseed was TCA-acetone method,and the protein spots on 2-DE map were the most.When IPG strip（pH 3-10）and 12% gel were used,and the loading amount was 250 μg,the two-dimensional electrophoresis map with the clear background,good repeatability and high protein spot resolution was obtained.[Conclusion] The research laid the foundation for carrying out the rapeseed proteomics research.

Preparation, friction and wear properties of hydrophobic lanthanum borate nanorods in rapeseed oil

Oleic acid (denoted as OA) surface-caped lanthanum borate nanorods, abbreviated as OA/LaBO3·H2O, were prepared via hydrothermal method. The microstructures of the as-prepared OA/LaBO3·H2O nanorods were characterized by X-ray diffraction (XRD) and scanning electron microscopy (SEM) techniques. The friction and wear properties of OA/LaBO3·H2O nanorods in rapeseed oil were evaluated with a four-ball tribo-tester. The results show that the as-prepared OA/LaBO3·H2O nanorods are hydrophobic and display nanorods morphology with uniform diameter of about 50 nm and length of up to 500 nm. In the meantime, OA/LaBO3·H2O nanorods can obviously improve the anti-wear and friction-reducing capacities of rapeseed oil, and the optimal anti-wear and friction-reducing properties of rapeseed oil were obtained at an OA/LaBO3·H2O content of 1% (mass fraction).

Cloning and Expression of a Profilin Gene from Rapeseed

Profilin has recently been identified as an actin-binding protein in higher plants. A cDNA clone (designated Repro) encoding profilin gene was isolated from rapeseed ( Brassica napus L. cv. canadian Tween) using RT-PCR technique. Sequence analysis showed 82% similarity to Zea mays L. ZmPro3, 85% to Arabidopsis AthPRF1, 82% to Nicotiana tabacum L. NTPRO, 81% to Oryza sativa L. profilin A. A new full-length cDNA was obtained by 5'-RACE and 3'-RACE techniques. Sequence analysis showed that the size of full-length cDNA is 672 bp which contains a major open reading frame of 134 amino, acids, 5' and 3' untranslated regions and a long Poly (A) tail. Northern blot analysis showed that the profilin gene is a pollen and anther specific gene.

Friction and wear performances of magnesium alloy against steel under lubrication of rapeseed oil with S-containing additive

A S-containing additive, sulfuration modified rapeseed oil （named as SRO）, was prepared by chemical modification of rapeseed oil with sulfur compounds. The results indicate that the friction and wear of the magnesium alloy-steel tribomates could be effectively reduced by formulating SRO into rapeseed oil lubricant. The friction coefficients and the wear volumes of magnesium alloy decrease with increasing contents of SRO. The surface lubricated with SRO-doped rapeseed oil was characterized by less wear as compared with that lubricated with neat rapeseed oil. The enhanced anti-wear and friction-reducing abilities of rapeseed oil by SRO in the lubrication of magnesium alloy against steel were ascribed to the formation of a composite boundary lubrication film due to the strong adsorption of SRO and rapeseed oil onto the lubricated surfaces and their tribochemical reactions with magnesium alloy.

Analysis of Seed-specificity of Silencing fad_2 Gene Expression in Transgenic Rapeseed Line W-4(Brassica napus L.)

This study was to investigate the efficiency and specificity of RNAi silencing on the expression of endogenous fad2 gene in transgenic line W-4. [Method] The relative expression of fad2 gene in seeds at different developmental stages of 7th, 14th, 21st and 28th day after flowering （DAF） as wel as the root, stem, leaf at winter seedling stages of both the transgenic line W-4 and non-transgenic control Westar by real-time fluorescence quantitative PCR. [Results] The results showed the relative expression of fad2 gene was gradual y increasing with the days after flowering in the seeds of the control Westar, while it was found decreasing significantly since the 21st DAF in the seeds of the line W-4. The decline was up to 60% in comparison with the control Westar. However, no significant difference in the relative expression of fad2 gene in other organs like root, stem and leaf was observed between transgenic line W-4 and non-transgenic control Westar. Fatty acid composition analysis showed the oleic acid desaturation parameter（ODP） in seeds of the line W-4 was 0.07 in average, decreased by nearly 75% than control Westar which was 0.24 in average, while no significant difference in the seedling root, stem and leaf was measured between transgenic rapeseed and control. [Conclusion] The results above validated that RNA interference in transgenic rapeseed W-4 is at a seed-specific manner, not interfering with fad2 gene expression in organs such as the root, stem and leaf. The study also found that the period of fad2 gene expres-sion decline was wel coincided with the expression of napin gene, both appeared at the 21st DAF, indicating that the expression of dsRNA of fad2 gene is precisely control ed by the napin promoter.

Effects of Interaction between Density and Nitrogen Rate on Growth and Yield of No-tilling and Direct Sowing Rapeseed in Chengdu Plain

[Objective] The aim was to explore the optimal density and nitrogen rate of no-tilling and direct sowing rapeseeds in Chengdu plain. [Methods] Effects of in- teraction between density and nitrogen rate on the growth and yield of direct sowing rapeseed under no-tillage condition were investigated with Chuanyou 58 as materials and a split-plot experiment adopted. [Results] In Chengdu Plain, the yields of rape- seed changed from increasing to decreasing with increase of density and nitrogen rate. Both of density and nitrogen rate had significant effects on growth and yield of rapeseed and the latter overweighed in the effect. In addition, interaction of the two had negative effects on rapeseed yield. The yield of rapeseeds achieved the highest at 3 395.25 kg/hm^2 with interaction of density at 30.00×10^4 plant/hm^2 and nitrogen rate at 180.00 kg/hm^2; the theoretical maximal yield was 3 403.41 kg/hm^2 with interaction of density at 40.80×10^4 plants/hm^2 and nitrogen rate at 198.90 kg/hm^2. [Conclusion] In Chengdu Plain, the optimal density and nitrogen rate are 30.00×10^4-45.00×10^4 plant/hm^2 and 180.00-198.90 kg/hm^2, respectively.

Fatty Acid Composition and Seed Quality Traits of the Transgenic Rapeseed W-4(Brassica napus L.) with Down-regulated Expression of fad2 Gene

To clarify the effect of down-regulated expression of fad2 gene on the seed nutritional quality of rapeseed, the fatty acid composition, amino acid composi- tion, oil content, protein content, crude fiber content and glucosinolate content in the seeds of both transgenic line W-4 and its control Westar were compared. The re- sults showed that the oleic acid content in W-4 was 86.03%±0.20%, which was 29.36% higher than that in the control （P〈0.01）; the linoleic acid content was 2.86%± 0.01%, which was reduced by 84.03% compared with that in the control （P〈0.01）; the linolenic acid content in W-4 was 3.04%±0.04%, reduced by 57.60% （P〈0.01）; the palmitic acid content in W-4 was 3.23%±0.07%, reduced by 18.63% （P〈0.01）; the eicosenoic acid content in W-4 was increased by 18.46% compared with that in the control （P〈0.01）; the erucic acid content in W-4 was increased by 13.15% （P〈 0.05）; there was no significant difference in stearic acid content between the treat- ment and control groups （P〉0.05）. The amino acid composition analysis showed that total 18 amino acids, including 8 essential amino acids, were detected in both W-4 and Westar; there were no significant differences in contents of the 18 amino acids between the treatment and control groups except that of tyrosine （P〉0.05）; the contents of oil, proteins, glucosinolates and crude fiber in W-4 were 45.40%± 0.17% （P〉0.05）, 18.18%±0.91% （P〉0.05）, 18.20%±1.21% （P〉0.05） and 12.29%± 0.04% （P〉0.05）, respectively. All the results above showed that the down-regulated expression of fad2 had great effects on fatty acid composition and accumulation in rapeseed seeds, but had no significant effects on other seed quality traits, such as oil content, protein content, crude fiber content and glucosinolate content.