The influence of Lactobacillus plantarum fermentation in selenium-enriched Brassica napus L.:changes in the nutritional constituents,bioactivities and bioaccessibility

Selenium(Se)-enriched Brassica napus L.is a valuable organic Se supplement.In this study,the fermentation broth enriched with organic Se(FFS)was prepared using Lactobacillus plantarum to ferment the substrate of Se-enriched Brassica napus L.Significant increases were observed after fermentation in total sugars,reducing sugars,soluble proteins,total phenolic content(TPC),and total flavonoid content(TFC).The organic Se was retained at a concentration of 54.75 mg/g in the freeze-dried sample.Principal component analysis and cluster analysis showed good separation between the FFS and unfermented(FS)groups.Fragrant 2-ethyloxetane had the highest content among all volatiles,while sinapine had the highest content among all phenolic compounds.The fermentation process showed remarkable improvement in the abundance and concentration of volatile compounds and phenolic contents,making FFS exhibit strong antioxidant activity and inhibitory capacity againstα-glucosidase activity.The bioaccessibility of phenolic compounds was significantly greater in FFS compared to FS.ADMET analysis revealed that the majority of phenolic compounds contained in FFS did not exhibit mutagenicity toxicity,hepatotoxicity,skin sensitization,or blood-brain barrier penetration,indicating a favorable level of biosafety.Overall,our study provides a new insight into the further utilization of Se-enriched Brassica napus L.in foods.

Landscape of Sequence Variations in Homologous Copies of FAD2 and FAD3 in Rapeseed(Brassica napus L.)Germplasm with High/Low Linolenic Acid Trait

Genetic manipulation(either restraint or enhancement)of the biosynthesis pathway ofα-linolenic acid(ALA)in seed oil is an important goal in Brassica napus breeding.B.napus is a tetraploid plant whose genome often har-bors four and six homologous copies,respectively,of the two fatty acid desaturases FAD2 and FAD3,which con-trol the last two steps of ALA biosynthesis during seed oil accumulation.In this study,we compared their promoters,coding sequences,and expression levels in three high-ALA inbred lines 2006L,R8Q10,and YH25005,a low-ALA line A28,a low-ALA/high-oleic-acid accession SW,and the wildtype ZS11.The expression levels of most FAD2 and FAD3 homologs in the three high-ALA accessions were higher than those in ZS11 and much higher than those in A28 and SW.The three high-ALA accessions shared similar sequences with the pro-moters and CDSs of BnFAD3.C4 and BnFAD3.A3.In A28 and SW,substitution of three amino acid residues in BnFAD2.A5 and BnFAD2.C5,an absence of BnFAD2.C1 locus,and a 549 bp long deletion on the BnFAD3.A3 promoter were detected.The profile of BnFAD2 mutation in the two low-ALA accessions A28 and SW is different from that reported in previous studies.The mutations in BnFAD3 in the high-ALA accessions are reported for thefirst time.In identifying the sites of these mutations,we provide detailed information to aid the design of mole-cular markers for accelerated breeding schemes.

BnaSD.C3 is a novel major quantitative trait locus affecting semi-dwarf architecture in Brassica napus L.

Plant height is a key plant architectural trait that affects the seed yield,harvest index and lodging resistance in Brassica napus L.,although the genetic mechanisms affecting plant height remain unclear.Here,a semi-dwarf mutant,df34,was obtained by ethyl methanesulphonate-induced mutagenesis.Genetic analysis showed that the semi-dwarf phenotype is controlled by one semi-dominant gene,which was located on chromosome C03 using a bulked segregant analysis coupled with whole-genome sequencing,and this gene was named BnaSD.C3.Then BnaSD.C3 was fine-mapped to a 297.35-kb segment of the“Darmor-bzh”genome,but there was no potential candidate gene for the semi-dwarf trait underlying this interval.Furthermore,the interval was aligned to the Zhongshuang 11 reference genome.Finally,combining structural variation analysis,transcriptome sequencing,phytohormone analyses and gene annotation information,BnaC03G0466900ZS and BnaC03G0478900ZS were determined to be the most likely candidate genes affecting the plant height of df34.This study provides a novel major locus for breeding and new insights into the genetic architecture of plant height in B.napus.

Detection of new candidate genes controlling seed weight by integrating gene coexpression analysis and QTL mapping in Brassica napus L.

Seed weight is a component of seed yield in rapeseed(Brassica napus L.).Although quantitative trait loci(QTL)for seed weight have been reported in rapeseed,only a few causal quantitative trait genes(QTGs)have been identified,resulting in a limitation in understanding of seed weight regulation.We constructed a gene coexpression network at the early seed developmental stage using transcripts of 20,408 genes in QTL intervals and 1017 rapeseed homologs of known genes from other species.Among the 10 modules in this gene coexpression network,modules 1 and 2 were core modules and contained genes involved in source–flow–sink processes such as synthesis and transportation of fatty acid and protein,and photosynthesis.A hub gene SERINE CARBOXYPEPTIDASE-LIKE 19(SCPL19)was identified by candidate gene association analysis in rapeseed and functionally investigated using Arabidopsis T-DNA mutant and overexpression lines.Our study demonstrates the power of gene coexpression analysis to prioritize candidate genes from large candidate QTG sets and enhances the understanding of molecular mechanism for seed weight at the early developmental stage in rapeseed.

Identification of a candidate QTG for seed number per silique by integrating QTL mapping and RNA-seq in Brassica napus L.

Seed number per silique(SNPS)is one of seed yield components in rapeseed,but its genetic mechanism remains elusive.Here a double haploid(DH)population derived from a hybrid between female 6Q006with 35–40 SNPS and male 6W26 with 10–15 SNPS was investigated for SNPS in the year 2017,2018,2019 and 2021,and genotyped with Brassica 60K Illumina Infinium SNP array.An overlapping major QTL(qSNPS.C09)explaining 51.50%of phenotypic variance on average was narrowed to a 0.90 Mb region from 44.87 Mb to 45.77 Mb on chromosome C09 by BSA-seq.Subsequently,two DEGs in this interval were detected between extreme individuals in DH and F_2populations by transcriptome sequencing at7 and 14 days after pollination siliques.Of which,BnaC09g45400D encoded an adenine phosphoribosyltransferase 5(APT5)has a 48-bp InDel variation in the promoter of two parents.Candidate gene association analysis showed that this InDel variation was associated with SNPS in a nature population of rapeseed,where 54 accessions carrying the same haplotype as parent 6Q006 had higher SNPS than103 accessions carrying the same haplotype as parent 6W26.Collectively,the findings are helpful for rapeseed molecular breeding of SNPS,and provide new insight into the genetic and molecular mechanism of SNPS in rapeseed.

Genome-wide and transcriptome-wide identification of the APX gene family in rapeseed (Brassica napus L.) and their expression features under low temperature stress

Ascorbate peroxidase(APX)is a crucial H2O2 scavenger that utilizes ascorbic acid as an electron donor and plays a significant role in plant stress resistance.This study aims to identify and characterize the Brassica napus L.APX gene family through genome and transcriptome sequencing,while also revealing their expression profile under low-temperature stress via transcriptome and proteome analysis.The results indicate the presence of 18 genes with three different conserved domains distributed in Brassica napus L.,which can be classified into three major branches based on phylogenetic analysis.Eleven members were predicted to have the low-temperature response component(LTR).Most APX genes exhibit up-regulated transcriptional expression under low temperature stress,particularly APX2,APX4,APX12,and APX18.In terms of proteomics data,only six members(APX2,APX4,APX8,APX12,APX17,and APX18)showed temporal specificity in their expression patterns.Therefore,this study provides valuable insights into the complexity of the APX family in the functional characterization of its genes for future research.

Genome-Wide Identification, Evolution and Expression Analyses of GA2ox Gene Family in Brassica napus L.

Gibberellin 2-oxidases(GA2ox)are important enzymes that maintain the balance of bioactive GAs in plants.GA2ox genes have been identified and characterized in many plants,but these genes were not investigated in Brassica napus.Here,we identified 31 GA2ox genes in B.napus and 15 of these BnaGA2ox genes were distributed in the A and C subgenomes.Subcellular localization predictions suggested that all BnaGA2ox proteins were localized in the cytoplasm,and gene structure analysis showed that the BnaGA2ox genes contained 2–4 exons.Phylogenetic analysis indicated that BnGA2ox family proteins in monocotyledons and dicotyledons can be divided into four groups,including two C_(19)-GA2ox and two C_(20)-GA2ox clades.Group 4 is a C_(20)-GA2ox Class discovered recently.Most BnaGA2ox genes had a syntenic relationship with AtGA2ox genes.BnaGA2ox genes in the C subgenome had experienced stronger selection pressure than genes in the A subgenome.BnaGA2ox genes were highly expressed in specific tissues such as those involved in growth and development,and most of them were mainly involved in abiotic responses,regulation of phytohormones and growth and development.Our study provided a valuable evolutionary analysis of GA2ox genes in monocotyledons and dicotyledons,as well as an insight into the biological functions of GA2ox family genes in B.napus.

Genome-wide association study exposed the pleiotropic genes for yield-related and oil quality traits in Brassica napus L.

Oilseed rape(Brassica napus L.) is an allotetraploid(AACC,2n=38) crop,valued for its edible oil and protein content.seed yield and nutritional composition of rapeseed are influenced by its yield and oil quality traits.However,the genetic basis of yield-related and oil-quality traits remain ambiguous.A panel of 266 diversified oilseed rape accessions was genotyped using 223 simple sequence repeat(SSR) markers covering all 19 chromosomes to identify significant markers associated with yield and quality traits.Twelve yield-related and six quality traits were investigated in two consecutive years(2014 and 2015),with three replications in two environments(Changshun,CS;and Qinghe,QH).Using the model GLM with population structure and kinship(Q+K),a total of 25 significant SSR markers(P <0.001) were detected to be associated with these twelve yield-related and six quality traits,explaining 4.56%-19.17% of the phenotypic variation for each trait.Based on these markers,BnaA03g23490D, BnaC09g46370D,BnaA07g37150D,BnaA01g32590D, and BnaC09g37280D were identified as pleiotropic genes controlling multiple traits.These candidate genes illustrated the potential for the genetic understanding of yield and oil quality traits.Most importantly,these significant markers can be used for marker-assisted breeding of oilseed rape in different environments.

甘蓝型油菜(Brassica napus L.)千粒重性状遗传体系分析

通过遗传差异较大的 2个甘蓝型油菜 (BrassicanapusL )纯系亲本组合 (HSTC14×宁油 7号 )衍生后代的世代家系群体分析 ,应用主基因 +多基因家系世代联合分离分析方法研究油菜千粒重的遗传体系。结果表明 ,甘蓝型油菜HSTC14×宁油 7号组合千粒重遗传体系系由一对主基因 +多基因构成 ,主基因中只有加性效应 (d =0 10 6 2 ) ,不存在显性效应 ;多基因存在加性 ([d]=0 2 2 2 8)和显性效应 ,但显性效应为负值 ([h]=- 0 2 80 0 )。表明该组合千粒重主要由主基因加性效应和多基因效应决定 ,但无杂种优势。F2∶3家系群体主基因遗传力为 83 39% ,多基因遗传力为 11 71%。HSTC14×宁油 7号组合千粒重的改良可能受主基因无显性效应 ,多基因显性效应为负值的制约。

锰胁迫对甘蓝型油菜Brassica napus L.种子活力和幼苗抗氧化系统的影响

用不同质量分数的锰溶液(0.2%,0.4%,0.8%和1.2%Mn2+)处理甘蓝型胜利油菜Brassica napuscv.Shengli,研究锰对其种子活力、幼苗细胞膜透性、脂质过氧化及抗氧化剂的影响.结果表明:锰胁迫降低了种子活力、幼苗抗坏血酸(Vc)质量分数、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性,破坏了细胞膜的完整性,并使丙二醛(MDA)质量分数、脯氨酸(Pro)质量分数和过氧化物酶(POD)活性增加.

Mechanical Harvesting Effects on Seed Yield Loss,Quality Traits and Profitability of Winter Oilseed Rape (Brassica napus L.)

China is one of the most important rapeseed producing countries in the world. Effective mechanical harvesting time for decreasing harvesting loss of winter oilseed rape has been becoming a critical factor. An elite cultivar Zhongshuang 11 （Brassica napus L.） was employed in two rounds of field experiments from 2009 to 2011. Seeds were sown with machine, three combine harvesting times namely combine harvesting A, B, and C （CHA, CHB, and CHC） were designed and manual harvesting （MH） as control was performed at maturity. The harvesting treatments were determined according to color of pod and seed in the field. Seed yield loss and quality in different treatments were evaluated. Results showed that both seed yields and harvesting losses in 2009-2010 were higher than that in 2010-2011, whereas seed oil contents in 2010-2011 were higher than that in 2009-2010. The highest yield appeared in CHB, which was significantly higher than that in MH. Furthermore, harvesting loss in CHB were 50% that in MH. Seed oil content and chlorophyll exhibited no obvious difference between CHB and MH. Economic profit analysis demonstrated that mechanical sowing/combine harvesting （MS/CH） showed an input/output ratio of 1：1.6, and it was 1：1.2 in mechanical sowing/manual harvesting （MS/MH）. Labor-cost accounted for more than 70% of the total cost in MS/MH, which led to low profitability to a great extent. Our results suggested that CHB was the optimum harvesting time for winter oilseed rape along the Yangtze River.

幼叶黄化油菜(Brassica napus L.)突变体Cr3529叶绿体超微结构观察

对油菜Cr3529叶片细胞中的叶绿体数目及结构进行了显微及亚显微观察.结果显示,突变与野生型细胞中叶绿体的数目没有显著性的差异,但叶绿体的结构有较大的差异.较之正常油菜,Cr3529油菜叶绿体基粒类囊体数较少,基质类囊体数较多,而且,在基粒类囊体中基粒片层数也较少,大部分基粒只有几个片层,基粒的平均片层数为5.46,约为野生型的一半.作者认为,较少的基粒片层数可能是引起Cr3529油菜叶绿素含量减少,并最终导致植株显著减产的重要原因.

Comparison of transcriptomes undergoing waterlogging at the seedling stage between tolerant and sensitive varieties of Brassica napus L.

RNA sequencing of the sensitive GH01 variety of Brassica napus L. seedling roots under 12 h of waterlogging was compared with previously published data of the ZS9 tolerant variety to unravel genetic mechanisms of waterlogging tolerance beyond natural variation. A total of 2 977 genes with similar expression patterns and 17 genes with opposite expression patterns were identiifed in the transcription proifles of ZS9 and GH01. An additional 1 438 genes in ZS9 and 1 861 genes in GH01 showed strain speciifc regulation. Analysis of the overlapped genes between ZS9 and GH01 revealed that waterlogging tolerance is determined by ability to regulate genes with similar expression patterns. Moreover, differences in both gene expression proifles and abscisic acid (ABA) contents between the two varieties suggest that ABA may play some role in waterlogging tolerance. This study identiifes a subset of candidate genes for further functional analysis.

抗倒伏甘蓝型油菜(Brassica napus L.)根和茎解剖学结构分析

对甘蓝型油菜(Brassica napus L.)6个抗倒伏材料和5个易倒伏材料的苗期和苔期的根部和茎部采用石蜡切片的方法切出6μm厚的薄片,用番红-固绿双重染色,借助Olympus显微镜观察不同材料茎的表皮、皮层、厚角组织、维管束和髄的结构,以及根的木质部、导管等,并对茎和根的结构特点进行了比较。结果表明:①在苗期的茎中,与易倒伏材料相比,抗倒伏材料有较多的厚角组织;抗倒伏材料维管束的面积占茎基部横切面面积之比比易倒伏材料平均高13.1个百分点;抗倒伏材料和易倒伏材料间维管束的个数差异不大,但抗倒伏材料的维管束排列相对比较紧密。②在苔期的茎中,抗倒伏材料的厚角组织和厚壁组织比易倒伏材料发达;虽然两者间维管束的个数和面积无明显差异,但抗倒伏材料维管束排列紧密。③在苗期的根部,抗倒伏材料和易倒伏材料木质部的面积占根部横切面面积之比差异不显著;但抗倒伏材料导管数目比易倒伏材料平均多74.7个,易倒伏材料中较大导管的数目多于抗倒伏材料。④在苔期的根部,抗倒伏材料木质部占根横切面面积的之比比易倒伏材料高8.33个百分点,抗倒伏材料单位面积导管的数目多于易倒伏材料。因此,较多的厚角组织和厚壁组织、较多的导管数和排列紧密的维管束是甘蓝型油菜抗倒伏材料苗期和苔期重要的解剖学特点。

Identification of a Cytoplasmic Male Sterile Line NEA in Brassica napus L.and Its Genetic Studies

Male sterile NEA plants were identified in progenies of the radiated 92P x Aggregate-silique in Brassica napus L. in 1993. Their progeny plants from test crossing and open pollination were 100% male sterile. The double-low male sterile lines JL-4 and JL-18 were bred through successive backcrossing of the double -low variety No.4 and No. 18 in Brassica napus L.to NEA. Restorer lines 6720 and 6722 with significant heterosis in F1 were developed. The results from investigating the restoring-ntaintaining relationship and inheritance of the restorer gene indicated that JL-4 and JL-18 were different from both PolCMS and Shan 2A type, and their restorer gene was controlled by a pair of dominant genie genes.

Detection of Tocopherol in Oilseed Rape (Brassica napus L.) Using Gas Chromatography with Flame Ionization Detector

The variation among Chinese genotypes of Brassica napus L. for seed tocopherols content and their analysis using gas chromatography has not been comprehensively reported till to date. In the present study, the tocopherol contents of four Chinese genotypes of Brassica napus L., namely, Gaoyou 605, Zhejiang 619, Zheshuang 758, and Zheshuang 72, were evaluated using three modified sample preparation protocols （P1, P2, and P3） for tocopherol extraction. These methods were distinguished as follows. Protocol one （P1） included the evaporation of solvent after extraction without silylation. Protocol two （P2） followed the direct supernatant collection after overnight extraction without drying and silylation. Protocol three （P3） included trimethylsilylation with N,O-bis（trimethylsilyl） trifluoroacetamide. Genotypic comparison of tocopherol and its isoforms revealed that Gaoyou 605 was dominant over the other genotypes with （140.5＋ 10.5）, （316.2＋ 9.2）, and （559.1＋ 24.3） ~tg g-~ of seed meal ct-, 7-, and total （T-） tocopherol, respectively, and a 0.44＋0.04 ^- to 7-tocopherol ratio. The comparison of the sample preparation protocols, on the other hand, suggests that P3 is the most suitable method for the tocopherol extraction from Brassica oilseeds and for the analysis of tocopherols using gas chromatography flame ionization detector （GC-FID）. Trimethylsilylation is the key step differentiating P3 from P1 and P2. Variations detected in tocopherol contents among the Chinese rapeseed （B. napus） genotypes signify the need to quantify a wide range of rapeseed germplasm for seed tocopherol dynamics in short and crop improvement in long.

QTL Analysis of the Oil Content and the Hull Content in Brassica napus L.

The QTLs of the oil content and the hull content were analyzed in Brassica napus L. by constructing the linkage map. The F2：6RIL population with 188 lines, derived from the cross of GH06 × P147, was used as the mapping population. The SRAP, SSR, AFLP, and TRAP markers were used to construct the linkage map, and the composite interval mapping （CIM） to identify the quantitative trait loci associated with the oil content and the hull content. 300 markers were integrated into 19 linkage groups, covering 1 248.5 cM in total. Seven QTLs were found to be responsible for the oil content with the single contribution to phenotypic variance ranging from 3.73 to 10.46%; four QTLs were found for the hull content with the single contribution to phenotypic variance ranging from 4.89 to 6.84%. The yellow-seeded Brassica napus L. has the advantage of higher oil content and the hull content has a significant effect on the oil content. In addition, the SRAP marker is good for detecting QTL.

甘蓝型油菜(Brassica napus L.)花药培养中若干因素对花粉胚状体诱导和植株再生的影响

研究表明,甘蓝型油菜(Brassica napus L.)花粉胚状体的诱导频率与培养材料的基因型、培养基蔗糖浓度、培养基附加成分、培养基状态、温度处理等因素有关。不同基因型材料的花粉胚状体诱导频率明显不同;培养基蔗糖浓度以10％为最佳;B_5培养基附加2,4-D 0.1mg/l和NAA0.1mg/l的培养效果最好;液体浅层静止漂浮培养优于固体培养;变温处理可以提高花粉胚状体诱导频率。本文也报道了五种不同的花粉胚状体类型与植株再生的研究。

Combining Ability and Genetic Effects of Germination Traits of Brassica napus L.Under Waterlogging Stress Condition

Cross combinations from six rapeseed cultivars and lines were evaluated under waterlogging stress condition in order to understand the genetic mechanism of waterlogging tolerance of Brassica napus L.and provide reasonable improvement programs.There were six germination traits investigated on combining ability and heritability using complete diallel crossing method designed for 30 combinations from those six cultivars and lines.The traits included relative root length,stem length,fresh weight per plant,survival rate,electrical conductivity,and vigor index.After flooding treatment,the six traits of parents and F1 were analyzed.The general combining ability（GCA） and special combining ability（SCA） of germination traits were analyzed using Griffing I method.Among 30 cross combinations,the GCA was significantly different among six waterlogging resistance traits.The SCA of these traits was significantly different except the SCA of electrical conductivity.As a representative trait of waterlogging tolerance in rapeseed,relative vigor index had the highest narrow heritability and relatively low broad heritability.The cultivars Zhongshuang 9 and P79 had higher tolerance potential to waterlogging stress.It can be concluded that combining ability and genetic effects of relative vigor index during germination stage could be used to identify the waterlogging tolerance of rapeseed in breeding program.

甘蓝型油菜(Brassica napus L.)无花瓣性状的遗传及其育种潜势

将正常品种与无花瓣亲本杂交,正反交F_1代中有瓣对无瓣为完全显性。在F_2及B_1中,有瓣:无瓣分别为255:1与15:1,表明无花瓣性状的遗传受着核中四对隐性基因p_1p_1p_2p_2p_3p_3p_4p_4的制约。无花瓣基因的育种价值主要体现在减耗节能,包括个体节能(减少建成花瓣的损耗)和群体节能(降低黄色冠层对自然光的吸收与反射损失),以及对菌核病的形态避病能力(田间发病率仅为1.75％)。