Identification of suitable reference genes in leaves and roots of rapeseed (Brassica napus L.) under different nutrient deficiencies

Nutrient deficiency stresses often occur simultaneously in soil. Thus, it＇s necessary to investigate the mechanisms underlying plant responses to multiple stresses through identification of some key stress-responsive genes. Quantitative real-time PCR （qRT-PCR） is essential for detecting the expression of the interested genes, of which the selection of suitable reference genes is a crucial step before qRT-PCR. To date, reliable reference genes to normalize qRT-PCR data under different nutrient deficiencies have not been reported in plants. In this study, expression of ten candidate reference genes was detected in leaves and roots of rapeseed （Brassica napus L.） after implementing different nutrient deficiencies for 14 days. These candidate genes, included two traditionally used reference genes and eight genes selected from an RNA- Seq dataset. Two software packages （GeNorm, NormFinder） were employed to evaluate candidate gene stability. Results showed that VHA-E1 was the highest-ranked gene in leaves of nutrient-deficient rapeseed, while VHA-G1 and UBC21 were most stable in nutrient-deficient roots. When rapeseed leaves and roots were combined, UBC21, HTB1, VHA-G1 and A CT7 were most stable among all samples. To evaluate the stabilities of the highest-ranked genes, the relative expression of two target genes, BnTrxl;1 and BnPhtl;3 Were further determined. The results showed that the relative expression of BnTrxl;1 depended on reference gene selection, suggesting that it＇s necessary to evaluate the stability of reference gene prior to qRT-PCR. This study provides suitable reference genes for gene expression analysis of rapeseed responses to different nutrient deficiencies, which is essential for elucidation of mechanisms underlying rapeseed responses to multiple nutrient deficiency stresses

Physiological and molecular responses to cold stress in rapeseed(Brassica napus L.)

Low temperature is one of the most important abiotic factors inhibiting growth, productivity, and distribution of rapeseed(Brassica napus L.). Therefore, it is important to identify and cultivate cold-tolerant germplasm. The objective of this study was to figure out the mechanism of chilling(4 and 2°C) and freezing(–2 and –4°C) stresses along with a control(22°C) in B. napus cultivars(1801 and C20) under controlled environment(growth chamber). The experiment was arranged in a complete randomized design with three replications. Our results exhibited that under chilling and freezing stresses, the increment of proline accumulation, soluble sugar and protein contents, and antioxidant enzyme activity were enhanced more in 1801 cultivar compared with C20 cultivar. At –2 and –4°C, the seedlings of C20 cultivar died completely compared with 1801 cultivar. Hydrogen peroxide(H2 O2) and malondialdehyde contents(MDA) increased in both cultivars, but when the temperature was decreased up to –2 and –4°C, the MDA and H2 O2 contents continuously dropped in 1801 cultivar. Moreover, we found that leaf abscisic acid(ABA) was enhanced in 1801 cultivar under chilling and freezing stresses. Additionally, the transcriptional regulations of cold-tolerant genes(COLD1, CBF4, COR6.6, COR15, and COR25) were also determined using real-time quantitative PCR(RT-q PCR). RT-q PCR showed that higher expression of these genes were found in 1801 as compared to C20 under cold stress(chilling and freezing stresses). Therefore, it is concluded from this experiment that 1801 cultivar has a higher ability to respond to cold stress(chilling and freezing stresses) by maintaining hormonal, antioxidative, and osmotic activity along with gene transcription process than C20. The result of this study will provide a solid foundation for understanding physiological and molecular mechanisms of cold stress signaling in rapeseed(B. napus).

Exogenous application of a low concentration of melatonin enhances salt tolerance in rapeseed(Brassica napus L.) seedlings

Melatonin is a naturally occurring compound in plants. Here, we tested the effect of exogenous melatonin on rapeseed（Brassica napus L.） grown under salt stress. Application of 30 μmol L^-1 melatonin alleviated salt-induced growth inhibition, and the shoot fresh weight, the shoot dry weight, the root fresh weight, and the root dry weight of seedlings treated with exogenous melatonin increased by 128.2, 142.9, 122.2, and 124.2%, respectively, compared to those under salt stress. In addition, several physiological parameters were evaluated. The activities of antioxidant enzymes including peroxidase（POD）, catalase（CAT） and ascorbate peroxidase（APX） were enhanced by 16.5, 19.3, and 14.2% compared to their activities in plants without exogenous melatonin application under salt stress, while the H2O2 content was decreased by 11.2% by exogenous melatonin. Furthermore, melatonin treatment promoted solute accumulation by increasing the contents of proline（26.8%）, soluble sugars（15.1%） and proteins（58.8%）. The results also suggested that higher concentrations（〉50 μmol L^-1） of melatonin could attenuate or even prevent the beneficial effects on seedling development. In conclusion, application of a low concentration of exogenous melatonin to rapeseed plants under salt stress can improve the H2O2-scavenging capacity by enhancing the activities of antioxidant enzymes such as POD, CAT and APX, and can also alleviate osmotic stress by promoting the accumulation of osmoregulatory substances such as soluble proteins, proline, and water soluble glucan. Ultimately, exogenous melatonin facilitates root development and improves the biomass of rapeseed seedlings grown under salt stress, thereby effectively alleviating the damage of salt stress in rapeseed seedlings.

Genetic Diversity of Chinese and Swedish Rapeseed (Brassica napus L.) Analyzed by Inter-Simple Sequence Repeats (ISSRs)

We have compared genetic diversity of 24 Chinese weak-winter, Swedish winter and spring B. napus accessions by inter-simple sequence repeats (ISSRs). By cluster analysis (UPGMA) based on 125 polymorphism bands amplified with 20 primers, the 24 accessions were divided into three groups. Six Swedish winter lines and eight Chinese weak-winter lines were in the group I and the groupⅡwere two Chinese weak-winter lines XiangyoulS and Bao81. The third group contained eight Swedish spring lines. Principal co-ordinates analysis (PCO) showed similar groupings to cluster analysis. Results from cluster analysis and PCO analysis showed very clearly that Chinese weak-winter, Swedish spring and winter accessions were distinguished from each other and Chinese weak-winter accessions in this study were genetically closer to Swedish winter accessions than to Swedish spring accessions. The Chinese weak-winter accessions had larger diversity than Swedish spring or winter accessions did. This study indicated that ISSR is a suitable and effective tool to evaluate genetic diversity among rapeseed germplasm.

Molecular evidence for blocking erucic acid synthesis in rapeseed(Brassica napus L.) by a two-base-pair deletion in FAE1(fatty acid elongase 1)

DNA sequences of fatty acid elongase 1 genes FAE1.1 （EA） and FAE1.2 （Ec） were isolated and characterized for 30 com- mercialized low erucic acid rapeseed （LEAR） cultivars in China. Four types of independent mutation leading to low erucic acid trait were found, i.e., a single-base transition （eAl）, a two-base deletion （ec2） and four-base deletion （eC4） as well as single-base transition with a four-base deletion （eA.）. Three genotypes, i.e., eA1eA1eC2eC2, eA1eA1eC4eC4 and eA.eA.ec4ec4 were responsible for LEA content in storage Iipids of different rapeseed cultivars. Most of the LEAR cultivars had a genotype of eA1eA1ec2ec2, which were descended from the first LEAR cultivar, Oro. Yeast expression analysis revealed that two-base-pair （AA） deletion （ec2） at the base sites of 1 422-1 423 in the C genome FAE1 gene resulted in the absence of the condensing enzyme and led to the failure to produce erucic acid. Coexpression of FAE1 and ketoacyI-CoA reductase （KCR） or enoyI-CoA reductase （ECR） was found in high erucic acid rapeseed （HEAR） but not in LEAR （eA1eA1ec2ec2oreA1eA1ec4ec4）. Moreover, KCR and ECR were still coordinately regulated in eA1eA1ec2ec2 or eA1eA1ec4ec4 genotypes, suggesting that the expression of two genes was tightly linked. In addition, specific detection methods were developed by high-resolution melting curve analysis in order to detect eA1 and ec4.

Relationship Among Yield Components and Selection Criteria for Yield Improvement in Early Rapeseed (Brassica napus L.)

The objective of this study was to determine the relationship between seed yield and other important agronomic traits of early-maturing rapeseed as a rotation crop in a double-cropping rice area using Pearson＇s correlation coefficient as well as to estimate direct and indirect effects of specific yield component traits on seed yield via path analysis. Nineteen rapeseed genotypes were grown at ten environments in South China during 2008-2009 and 12 characters were evaluated. Analysis of variance showed that environment had a significant impact on all characters. For most characters the genotype by environment interaction was weak and not statistically significant. Simple correlation analysis indicated that the number of primary branches （PB）, number of pods on the main raceme （PR）, and number of seeds per pod made significant contributions to seed yield per plant （SYP）, while 1 000-seed weight was negatively correlated with SYP （r=-0.485, P0.05）. Furthermore, number of pods per plant （PP）, PB, and PR had the greatest direct effects on SYP. In addition, PP and PB were the best indicators to predict seed yield in stepwise regression analysis. Finally, yield component differences between early- and medium-maturity varieties were compared; this showed that to improve the seed yield of early varieties, more emphasis should be given to increase PP, PB, and PR, and reduce plant height and shortening of growth duration in breeding practice.

Heterosis for Seed Yield, Oil Content and Other Characters in Rapeseed (Brassica napus L.)

Nine inbred lines of rapeseed （Brassica napus L.） used as male were crossed to five recessive genetic male sterile （RGMS） lines used as female to produce 45 single crosses. The crosses, their parents and a check hybrid were tested at two locations during 2007 to 2008 for testing the performance and heterosis of hybrids for seed yield and other characters. Results showed that variations for seed yield, oil content, days to flowering and days to maturity were significant. Mean squares for hybrids were significant for all characters. High heterosis （-4.5%-88.3%）, heterobeltiosis （-15.6%-81.1%） and standard heterosis G34.8%-33.1%） were found for the seed yield. The highest heterosis and heterobeltiosis were found in the cross QH303-4A× 1190. The highest standard heterosis was found in the cross Qianyou8A×Q034. Both positive and negative heterosis of single crosses were detected for the oil content. Small heterosis was found for days to flowering and days to maturity. Among parents, Ⅲ 188, Ⅲ224, and Q034 were proved to be the superior for seed yield when used as parents in most of the hybrid combinations. 2365, Ⅲ224, and QH303-4AB were good for high oil content breeding, Ⅲ 176, 2313 and Qianyou3AB were good for the early hybrid breeding. Total 11 hybrids yielded higher than 2 500 kg·hm^-2 and also gave significantly positive heterosis, heterobeltiosis and standard heterosis. Among them, 10 crosses gave higher oil content than that of check. These crosses can be used in the future breeding program for the seed yield and the oil content. Two crosses including Qianyou3A × Ⅲ224 and Qianyou3A×2313 can be used for the early breeding program.

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.

Inheritance of Erucic Acid,Glucosinolate,and Oleic Acid Contents in Rapeseed (Brassica napus L.)

This study was conducted to verify the inheritance of certain characters of rapeseed including erucic acid, glucosinolate and oleic acid contents by using generation mean analysis. The cross of lines Ⅲ174×Zi20 （F1）, F2, BC1 （F1×P1）, BC2 （F1×P2）, and parents （P1 and P2） were evaluated in the field. Data were measured on individual plants for oleic acid, erucic acid, and glucosinolate contents. Transgressive variations in F2 population were observed for oleic acid content, indicating that dominance and recessive genes distributed in both parents. Scaling test indicated that the effects of genes controlling these characters did not follow the additivedominance model. The data for three characters were analyzed using six parameter models and found that one or more types of epistatic gene effects were important for glucosinolate content. High broad sense heritabilities were obtained for erucic acid, oleic acid, and glucosinolate contents with the values of 98.97%, 93.68%, and 86.17%, respectively. Two major gene pairs were found to control the expression of erucic acid and oleic acid contents, while three major gene pairs were detected to control glucosinolate content.

Physiological and anatomical changes in two rapeseed (Brassica napus L.) genotypes under drought stress conditions

Understanding the mechanisms of drought resistance in crop species is crucial for the selection and breeding of tolerant rapeseed(Brassica napus L.)varieties.The present study aimed to assess the physiological and anatomical responses of two rapeseed genotypes,P287(drought-tolerant)and T88(drought-sensitive)under three intensities of drought stress.All physiological and anatomical parameters related to drought acclimation were significantly altered in both genotypes under stress conditions.At the fourth-leaf stage,the relative water content,chlorophyll content,protein content,malondialdehyde content,and the activities of peroxidase and catalase in P287 were significantly higher than those in T88,particularly under severe drought conditions.After rehydration,all physiological indexes recovered rapidly,especially in P287.In addition,under drought stress,compared with T88,P287 had thicker palisade tissue,thinner spongy tissue,higher ratio of chloroplast length to chloroplast width,higher stomatal density and stomatal closure rate.Overall,the interaction between physiological and anatomical features improved the drought tolerance of P287 under drought stress conditions.

Identification and evaluation of high nitrogen nutrition efficiency in rapeseed(Brassica napus L.)germplasm

To establish identification and evaluation methods of N(nitrogen)absorption and utilization of rapeseed(Brassica napus L.),difference of N nutrition efficiency(NNE)among rapeseed germplasms and relationship between NNE and plant traits under various N application rates were analyzed in this research.Pot cultivating experiments were conducted to investigate NNE with 3 N application rates in soil(0.05,0.2 and 0.3 g/kg).A total of 12 rapeseed germplasms were planted,nitrogen absorption efficiency(NAE)and nitrogen utilization efficiency(NUE)in seedling stage,bolting stage,initial flowering stage,final flowering stage,and maturity stage were obtained.Results showed that bolting stage was the best period for NAE identification and evaluation.Low N application rate in soil(0.05 g/kg)was the best for NAE,and the indirect indexes were basal stem diameter,plant root dry weight and above ground plant dry weight.Maturity stage was the best period for NUE identification and evaluation.High N application rate in soil(0.3 g/kg)was the best for NUE,and indirect indexes were above ground plant dry weight and basal stem diameter.N application rates of 0.05 g/kg in soil was the best for nitrogen harvest index at maturity stage,and indirect indexes was number of pods per plant.Plant traits of rapeseed germplasms affected NNE.Higher basal stem diameter,plant root dry weight and above ground plant dry weight at bolting stage under low N application rate were important characteristics of N absorption in rapeseed.Higher above ground plant dry weight and basal stem diameter at maturity stage under high N application rate were important characteristics of N utilization.Higher number of pods per plant at maturity stage under lowe N application rate was an important characteristic of N harvest index.These results provided a reliable index for N management and provided theoretical basis for guiding rapeseed breeding.

Foliar K application delays leaf senescence of winter rapeseed(Brassica napus L.) under waterlogging

To better understand waterlogging effect on leaf senescence in winter rapseed (Brassica napus L.) during flowering stage, experiments were designed to explore foliar K application influences on adverse effects of waterlogging stress. Winter rapeseed was sprayed with K after waterlogging at initial flowering stage. Results indicated that waterlogging significantly decreased leaf net photosynthetic rate (Pn), stomatal conductance (Gs), intercellular CO2 concentration (Ci) and transpiration rate (Tr). It also declined maximum quantum yield of PS II (Fv/Fm), quantum yield of electron transport (ΦPS II) and photochemical quenching (qP), but increased leaf non-photochemical quenching (NPQ) and minimal fluorescence (Fo). Interestingly, exogenous application of K significantly alleviated waterlogging-induced photosynthesis inhibition. Foliar K application increased RuBisCO activation, chlorophyll and soluble protein contents, while significantly decreased MDA content under waterlogging stress. Moreover, K supplementation improved accumulation of K+, Ca2+, Mg2+, N, Zn2+, Mn2+, Fe2+ in leaves. In general, foliar K application is effective inalleviating deleterious effects of waterlogging stress and delays leaf senescence of winter rapeseed.

Dissecting the genetic architecture of glucosinolate compounds for quality improvement in flowering stalk tissues of Brassica napus

Glucosinolates(GSLs) and their hydrolytic products contribute to the quality traits of rapeseed flowering stalk tissues, such as taste, flavor and anticarcinogenic properties(Glucoraphanin). However, little is known about the genetic mechanisms of GSL accumulation in rapeseed flowering stalks. In this study, the variation and genetic architecture of GSL metabolites in flowering stalk tissues were investigated for the first time among a panel of 107 accessions. All GSL compounds exhibited continuous and wide variations in the present population. Progoitrin,glucobrassicanapin and gluconapin were the most abundant GSL compounds. Five quantitative trait loci(QTL) significantly associated with three GSL compounds were identified by genome-wide association study. GRA_C04 was under selected during modern breeding, in which the ratio of lower GSL haplotype(HAP2) in the accessions bred before 1990(52.56%) was significantly lower than that after 1990(78.95%). Four candidate genes, BnaA01. SOT16, BnaA06. SOT17, Bna A06. MYB51a, and Bna A06. MYB51b, were identified in the GTL_A01 and 4OH_A06 regions.These findings provide new insights into GSL biosynthesis in flowering stalk tissues and facilitate quality improvement in rapeseed flowering stalks.

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.

Recent Advances,Problems and Outlooks in Rapeseed(Brassica napus L.)Breeding in China

As a big rapeseed(Brassica napus L.) producing and consuming country, China provides more and more rapeseed oil year by year. With the development of demand for rapeseed and import of edible oil, rapeseed breeding and production is facing a great challenge. New situation has proposed higher requirements to yield, oil quality and safety, etc. In the review, some aspects about rapeseed breeding in China were reported, including breeding of high-oil content rapeseed, breeding of extremely early-maturing rapeseed, development of mechanization and development of chemical emasculation. Some prospects and expectations were also proposed.

Analysis on Combining Ability for Characters of Male Sterile Lines in Rapeseed(Brassica napus L.)

The male sterile line is very important in the hybrid breeding program of rapeseed. This study was conducted to evaluate the combining ability of many characters of male sterile lines in Brassica napus L. Ten recessive genetic male sterile （RGMS） lines were used as parents to produce 45 single cross hybrids by using a half diallel cross method. These 45 crosses and their 10 parents were evaluated at Guiyang during 2007-2008. The results showed that both general combining ability （GCA） and specific combiing ability （SCA） effects were important for all characters, but additive gene effects were more predominant than non-additive gene effects. Qianyou 8AB and You 2894AB gave respective highly significant GCA effects of 230.94 and 127.65 kg-hm-2 for seed yield. Lines You 2894AB, QH303-4AB, You 157AB and You 2341AB gave highly significant GCA effects for oil content of 0.99, 1.62, 1.20 and 1.53%, respectively. The crosses among lines Qianyou 3AxQianyou 8B, Qianyou 8AxYou 2894B, You 2894AxQianyou 6B, Qianyou 8AxQH303-4B and Qianyou 8Ax Qianyou 6B gave high SCA effects of 616.29, 398.71,356.48, 394.24 and 303.79 kg hm-2 for seed yield, respectively. All these crosses also gave high seed yield indicating that these crosses could be used in the breeding program.

Worldwide rapeseed (Brassica napus L.) research:A bibliometric analysis during 2011–2021

Rapeseed(Brassica napus L.) is the world?s second produced oilseed and accounts for nearly 12% of world major vegetable oil production.For the last 10 years,the production,planting area,and yield of rapeseed have been stable,with improvement of seed quality and especially seed oil content.This paper presented the worldwide rapeseed research by using the bibliometric analysis of papers published during the period 2011–2021.In total,7617 articles and reviews were included in this analysis.Our results showed that the global publication on rapeseed increased substantially in the past 11 years,and the number of publications of China grew significantly in particular.China was the largest contributor to rapeseed research with higher H-index value and number of citations compared to the other countries.China also had the largest number of leading universities and institutions,of which Huazhong Agricultural University was the most productive.The scientific mapping including co-authorship network of countries/regions and authors,and co-occurrence network of author keywords were analyzed using VOSviewer software.The most common rapeseed research topics included oil,rapeseed meal,yield,biodiesel,fatty acids,salt stress,and proteins,while the current research focused on topics such as:abiotic stress,evolution,expression analysis,phylogenetic analysis,heterosis,polyploidy,and transcriptomics.This paper depicts the knowledge structure of current global rapeseed research that may help direct the future studies for relevant researchers.

A review: status and prospect of heterosis prediction in rapeseed(Brassica napus L.)

Heterosis has been widely exploited and utilized in rapeseed breeding. Reliable and precise prediction techniques of heterosis contributed in accelerating the crossbreeding and reducing the cost of large-scale field evaluation. Combined with our work in last decades, this review summarized the progress on heterosis prediction of rapeseed (Brassica napus L.), compared the efficiency of different methods, and proposed its prospect.

BnbHLH92a negatively regulates anthocyanin and proanthocyanidin biosynthesis in Brassica napus

Yellow seed trait is a desirable characteristic with potential for increasing seed quality and commercial value in rapeseed,and anthocyanin and proanthocyanidins(PAs)are major seed-coat pigments.Few transcription factors involved in the regulation of anthocyanin and PAs biosynthesis have been characterized in rapeseed.In this study,we identified a transcription factor gene BnbHLH92a(BnaA06T0441000ZS)in rapeseed.Overexpressing BnbHLH92a both in Arabidopsis and in rapeseed reduced levels of anthocyanin and PAs.Correspondingly,the expression profiles of anthocyanin and PA biosynthesis genes(TT3,BAN,TT8,TT18,and TTG1)were shown by quantitative real-time PCR to be inhibited in BnbHLH92a-overexpressing Arabidopsis seeds,indicating that BnbHLH92a represses the anthocyanin and PA biosynthesis pathway in Arabidopsis.BnbHLH92a physically interacts with the BnTTG1 protein and represses the biosynthesis of anthocyanins and PAs in rapeseed.BnbHLH92a also binds directly to the BnTT18 promoter and represses its expression.These results suggest that BnbHLH92a is a novel upstream regulator of flavonoid biosynthesis in B.napus.

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.