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  • 2013 Volume 35 Issue 5
    Published: 28 October 2013
      

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  • CUI Jia-cheng,MEI De-sheng,LI Yun-chang,LIU Jia1,FU Li,PENG Peng-fei,WANG Jun,WANG Hui, HU Qiong*
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    To understand the genetic effects of silique related traits on silique shatter resistance of oilseed rape, an incomplete diallel cross with 11 parental lines and 30 F1 crosses were made for multivariable conditional analysis at Xinzhou and Hanchuan counties of Hubei Province. Results showed that the largest contribution rate of phenotypic (28.2%) and additive (56.8%) effect to silique shatter resistance index (SRI) variance was from silique wall weight, the second was from silique length, with contribution rates of 17.5% and 33.4% for phenotypic and additive effects to SRI variance respectively. The investigated silique related traits had low contribution rates of dominant effect to SRI variance. The contribution rate of dominant × environment interaction effect of silique number per centimeter to SRI variance was the highest (44.7%) among silique related traits. The silique trait with the highest contribution rate of additive effect to silique SRI variance varied depending on different parental lines, indicating that the parental lines, especially those with relatively high silique SRI, possess different genetic backgrounds. It suggested that the strategies for improving shatter resistance included utilizing wild backgrounds of parents with large additive effects, and hybrids from multiple crosses with high contribution traits to increase resistance.

  • PU Hui-ming, LONG Wei-hua, GAO Jian-qin, HU Mao-long
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    Silique shatter strength (SSS) of 286 Brassica napus varieties (lines) was measured using a modified fracture method. Results indicated a wide variation in silique shattering resistance among these accessions. Among 146 hybrid varieties’ SSS values, 3 (2.05%) were less than 1N, 96 (65.75%) were between 1 to 2N, 47 (32.19%) were between 2 to 3N, no hybrid was found having more than 3N of SSS. The SSS of hybrids ranged between 0.82 to 2.87N, with an average of 1.83N, and CV (coefficient of variation) of 22.11%. Among 140 conventional varieties, 6 (4.29%) were less than 1N, 76 (54.29%) were between 1 to 2N, 49 (35%) were between 2 to 3N, and 9 (6.42%) were more than 3N. The SSS of conventional varieties ranged between 0.58 to 3.47N, with an average of 1.94N, and CV of 31.55. Correlations between SSS and silique- and seed-related traits showed that the SSS had highly significant positive correlations with single silique weight, unit area weight of silique wall, and 1 000-seed weight. And it had significant positive correlations with seed per silique and silique width. Analysis of the relations between hybrids and its restorers illustrated that the hybrid SSS had significant positive correlations with silique width, single silique weight, unit area weight of silique wall, and 1 000-seed weight of the restorer. Results indicated that silique size affected shattering trait. Rapeseed with large silique, thick wall and heavy seeds might have higher resistance to silique shattering. And large silique and heavy seeds of restorer lines would provide greater benefit to the shattering resistance of hybrids.

  • ZHAO Hui-Yan, XIAO Lu*, DU De-Zhi
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    Rape Dahuang is a special landrace of Brassica rapa L. which originated from Qinghai-Tibetan Plateau with bright yellow seeds. Previous results showed the yellow-seeded trait in Dahuang was controlled by a recessive gene (Brsc1), which was mapped on A09 in B. rapa. For more specific markers linked to Brsc1, BC1 population and F2 population were constructed from the cross of Dahuang and 09A-126 (brown-seeded, B. rapa). Amplified fragment length polymorphism (AFLP) technology combined with bulked segregant analysis (BSA) was conducted by screening 256 pairs of AFLP primers in BC1 population, 5 markers tightly linked to Brsc1 were obtained. 5 specific AFLP fragments were cloned, sequenced and identified, and compared to that of synteny in B. rapa genome, showing homology to A09 of B. rapa. The AFLP markers were converted into 5 SCAR (sequence characterized amplified region) markers for F2 population. One co-dominant marker was detected.

  • YAN Ming-li, LIU Li-li, XIANG Jian-hua, DENG Su-ping, SHU Jia-bin, SUN Chan
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    ANS genes were cloned from Brassica nigra by homology-based clone strategy. 2 copies named BnANS1 and BnANS2 were obtained. BnANS1 sequence was 1 366bp with a 76bp-intron. BnANS2 sequence gene was 1 367bp with 76bp intron. Sequence analysis showed polymorphisms in coding regions, intron, 5'- and 3'-noncoding regions of the two genes, including 3 polymorphic bases and a 4-base deletion in 5'-noncoding region, 26 polymorphic bases in coding region, 2 polymorphic bases in intron, 18 polymorphic bases and a 3-base insertion in 3'-noncoding region. The two copies encoded deduced a polypeptide of 356 amino acids, having the same conserved domain of ANS as other species, belonging to 2OG-Fe (II) oxygenase superfamily. BnANS1 protein was predicted to be 40 448.58Da with pI of 5.05, while BnANS2 was 40 468.52Da with pI of 5.04. Nine polymorphisms were found in their protein sequences. RT-PCR result showed that BnANS1 and BnANS2 expressed in seed coats and embryos in B. nigra. Compared with published ANS gene sequences of Brassica species, a pair of primer was obtained based on B genome-specific nucleotide variations loci, which could effectively identify B-genome origin of ANS in Brassica by allele-specific PCR. 

  • LI Qiang, SU Er-hu, GAO Ju-lin*, SUN Ji-ying, YU Xiao-fang, XIE Min
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    Start codon targeted polymorphism (SCoT) is a novel gene targeted marker technique. L25(56) orthogonal experiment and single factor experiment design were applied to optimize SCoT-PCR amplification system of soybean in five factors such as Mg2+, dNTPs, primer, Taq DNA polymerase and template DNA. The results showed that the optimized system was as follows: a total volume of 20μL including 2.0 mmol•L-1 Mg2+, 0.2 mmol•L-1 dNTPs, 0.250μmol•L-1 primer, 1.5U Taq polymerase, 30ng DNA template. The most suitable annealing temperature of primers was 50.4℃. The optimized SCoT-PCR system was tested on eighteen soybeans, and the result was stable and reliable. From the 82 primer combinations tested, 32 were selected with clear band patterns and abundant polymorphism. Amplifications and genetic verification were carried out on Jiyu 75, local black soybean and progenies from Jiyu 75×local black soybean. The results showed that the site in all progenies were derived from their parents and had their parents’ deletion. The system provided a new technology for evaluation of genetic diversity, construction of genetic linkage map and molecular marker assisted breeding for the soybean.

     

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    (1.Guangxi University, Nanning 530004; 2Cash Crops Research Institute of Guangxi Academy of Agricultural Sciences, Nanning 530007; 3Guangxi Crop Genetic Improvement and Biotechnology Lab, Nanning 530007)

    Abstract: Interspecific hybridization between a tetraploid cultivated peanut Zhongkaihua No.4 as female and one wild peanut species A. chacoensis as male was used to study the botanical characteristics and pollen fertility of their F1, synthetic allohexaploid S0 and selfing generations (S1~S3), and their genome changes were studied by SSR markers. The variation coefficients of total number of branches, No.1 lateral branches and corolla width of F1~S3 were 48.48%, 34.56% and 13.74% respectively. The result showed instability in early generations. Furthermore, the pollen germinated capability of F1~S3 was 0, 11.03%, 10.58%, 16.44% and 18.53% respectively, which demonstrated that fertility increased with the self-cross course. The amplified microsatellite bands began to alter as early as in F1 generation and the changes included the loss of parental fragments, simultaneous appearance of parental bands and appearance of novel fragments. The microsatellites and their flanking areas demonstrated drastic and rapid variations, which suggested their biological function could be related to polyploidy evolution.

  • LIU Hua, ZHANG Xin-You, HAN Suo-yi, YAN Mei, XU Jing, Dong Wen-zhao,Sun Zi-qi
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    Main stem height and lateral branch length of peanut are two important traits controlling plant shape. Recombinant inbred lines (RILs, Zheng9001×Zheng8903) were planted in Sanya, Hainan Province and Yuanyang, Henan Province. Main stem height and lateral branch length in the two environments were analyzed with mixed major gene and polygene inheritance model. Composite Interval Mapping (CIM) implemented in WinQTLcart2.5 was used to identify QTL of these two traits. The results indicated that the genetic models of both traits in Hainan were consistent with model C (C-0), the multi-gene genetic model with multiple gene effects and environmental effects, and the polygenic heritability of main stem height and branch length were 46.18% and 50.14% respectively. The genetic model of main stem height in Yuanyang was consistent with model E-1-0, two additive epistatic major genes-additive epistatic polygene model, and the genetic heritability of major genes and polygene were 45.49% and 21.98% respectively. The genetic model of branch length in Yuanyang was the same as that in Sanya, and the genetic heritability was 88.19%. Four and six QTL controlling main stem height were detected based on the phenotypic data collected in Sanya and Yuanyang respectively, and their phenotypic variance was 5.81%-18.00%. Five and eight QTL controlling branch length were detected based on data collected in Sanya and Yuanyang respectively, and their phenotype variance was 5.61%~25.12%. Based on the present study, it could be concluded that main stem height and branch length were controlled by polygene, which should be taken into account in the genetic improvement of plant shape.

  • GE Yu-bin, CHEN Bing-dong, MAO Xu-hui, JIA Xiu-ping
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    Seven cytoplasmic male-sterile and six-branched male restorer lines of the improved oil sunflower were hybridized with 7×6 incomplete diallel, 42 F1 hybrids were further used to estimate their general combining ability (GCA)、specific combining ability (SCA) and total combining ability (TCA) effects on main economic traits of oil Sunflower. The components of variance for combining ability revealed that mean squares were highly significant for all of the cross combinations, showed that the preponderance of additive gene action played the leading role in leaves per plant, kernel oil content and kernel rate, and indicated the preponderance of non-additive gene action was predominant in plant height, head diameter and growth duration, while seed yield per plant and 1 000-seed weight were attributed to additive gene action and non-additive gene action variance. GCA effects of the parents revealed that among the CMS LQ194A and LQ252A and among the restorers Y109, 9706R, F15-1R were found to be promising parents. Based on significant SCA and TCA effects in components analysis, seven hybrids LQ218×9706R, LQ28×Y109, LQ28×9706R, LQ178×Y109, LQ218×F15-1R, LQ28×F15-1R and LQ28×Y112 were identified as distinct for comprehensive traits and heredity characteristics. The CMS played a main role in the formation of the F0 traits of the head diameter and 1 000-seed weight. The result of correlation analysis indicated a significant negative correlation between 1 000-seed weight and kernel rate and a highly significant positive correlation between 1 000-seed weight and seed yield per plant. The phenotypic correlations and genotypic correlations were consistent.

  • HUANG Xian-qun, Francoise Fabre, Ahmad Sarraffi, LIU Zhu-zi, Laurent Gentzbittel
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    In an attempt to locate the somatic embryogenesis genes and select their relevant markers for marker-assisted selection in sunflower, 31 recombinants inbred lines F8 (RILs) derived from the cross between PAC-2 and RHA-266 were investigated. Somatic embryogenesis was induced using the thin epidermal layers from seedling hypocotyls. The analysed result, the genetic map, and 3 QTLs loci associated with number of epidermal embryogenesis per 100 epidermis cultured (EE/EC%) were located at 5, 10, 13 linkage groups. They could explain the phenotypic variation of 14.61%、10.04% and 14.07% respectively. In addition, 9 QTLs loci associated with number of embryos per epidermal embryogenesis (E/EE) were located at 2, 8, 10, 11, 12 and 19 linkage groups. They could explain the phenotypic variation of 6.64%~16.96%. Four QTL loci of them were located at some 10 linkage group and their positions were adjacent from 34.57 cM to 47.10 cM. They could explain the phenotypic variation of 40.39%. In order to verify the results obtained and to increase the density of marker in the mapping of the QTL regions for selecting their relevant markers, the parameter of EE/EC% was determined by the following test. Two F2 segregation populations were obtained by cross between 2 RILs with one high somatic embryogenesis and one low each. AFLP and SSR markers were used to scan these two F2 populations. There were 24, 6 and 18 new markers inserted to 5, 10, 13 linkage groups respectively, and 6, 0 and 4 new markers correlated with the somatic embryogenesis added to their QTL regions respectively. The intervals of these three linkage groups were shortened from 6.80-11.40 cM to 5.10-5.60 cM. The genotypes of the F2 lines were compared to and analyzed with their parents and grand-parents in QTL regions of EE/EC%. Seven lines with a high somatic embryogenesis capability and 7 lines with a low capability were preselected. The induced somatic embryogenesis of F3 families tested and verified the preselected results. Results showed that the predicted results were consistent with actual results. Conclusion: the genes controlling E/EE were located mainly in 10 linkage group. The genes controlling EE/EC% were located mainly in 5 and 13 linkage groups. Four markers (e32m50-1 e38m49-2, e40m49-1, e32m62-10) were the closest markers to the genes of somatic embryogenesis. They could be used in marker-assisted selection. 

  • BAO Hai-zhu, GAO Ju-lin, MA Qing, HU Shu-ping
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    The additive-dominant-maternal model with genotype × environment interaction was analyzed by both unconditional variance and conditional variance separately. The seed oil contents in oil sunflower were estimated at different stages in different environments. The estimations of genetic effect for oil content of seed were determined at the developmental stage(0→t)or the special developmental period(t-1→t). The results showed that the oil content of seed was controlled by both dominant effect and maternal effect. The expression differences of seed nuclear genes and diploid maternal plant genes were observed on oil deposit during different developmental stages. The additive effect and the maternal effect were expressed inconsistently. The additive effect had not been detected at 0-10d and 30-40days after flowering, but it had been detected at peak during the days of 20-25d. There were highly significant dominant effects during 0-45 days after flowering, especially during the period of 15-30 days. It was the predominant stage of heterosis performance, while the maternal effect was active from 35 to 45 days after flowering, especially during the 35-40 days. The expression of genetic effect was easily affected by environment in early and late stages of flowering.

  • LIU Wen-ping,REN Guo-xiang, WEN Fei
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    To characterize the genetic diversity of the sesame germplasms, Ten amplified fragments length polymorphism (AFLP) primer combinations were used to screen 98 sesame germplasms from Shanxi Province and the major sesame planting areas such as Henan, Anhui, Hubei and Jiangxi provinces. These AFLP primer combinations produced 273 DNA bands in total with 76.9% polymorphic, and their polymorphic information content (PIC) varied from 0.164 to 0.271 with an average of 0.210. Shannon’s information index of different province germplasms groups were 0.091 (Jiangxi province) to 0.454 (Shanxi province). Similarity coefficients of the 98 sesame germplasms ranged from 0.280 to 0.996 with average of 0.798, and the germplasms from Shanxi province had the widest variation. The unweighted pair group method with arithmetic averages (UPGMA) cluster analysis for the 98 germplasms could distinguish the Shanxi sesame germplasms clearly from other provinces. Structure analysis inferred two clusters, and one of the clusters consisted of Shanxi sesame germplasms. These results indicated that the sesame germplasms from Shanxi Province had greater genetic diversity than those from the major planting provinces, and we should focus on the sesame germplasm collecting from minor sesame planting areas in future.

  • JIANG Lei,LI Gang,YUE Shuai, LIU Hong, YAN Xing-chu, QIN Rui*
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     11 Carthamus tinctorius L. varieties were used to study their genetic diversity and relationship using SRAP. With 25 SRAP primers,308 bands were amplified,among them 109 bands (35%) were polymorphism. The genetic similarity coefficients ranged from 0.82 to 0.9342 with an average of 0.87, indicating the existence of genetic diversity among these 11 varieties which were divided into four groups by cluster analysis at the similarity coefficient of 0.8914.

  • LI Jun, CHAO Ying, GUAN Chun-yun, MA Ni, YUAN Jin-zhan, REN Li, FANG Xiao-ping, LI Guang-ming, ZHANG Chun-lei*
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    A pot experiment was conducted to investigate physiological indexes of rapeseed (Brassica napus L. cv Zhongshuang 11) 0 to 24h after daubing leaves with methyl viologen (MV). Results indicated that antioxidant enzymes activity, O2- generation rate and MDA content were increased, while Pn and the content of soluble sugar were decreased, during the incipient stage of oxidation stress. Antioxidant enzymes activity, O2- generation rate, content of MDA and soluble sugar increased at the beginning then decreased. SOD activity reached the peak value of 485.16U•g-1FW at 4h, while activity of POD and CAT, O2- generation rate and MDA content went to the peaks at 6h, which increased 69.48,62.81%,98.65% and 79.17% respectively compared to CK. Regression analysis indicated that physiologic factors as SOD, POD, CAT, MDA and soluble protein influenced the high photosynthetic rate in rapeseed under short oxidant stress in this experiment, and CAT was the most important factor.

  • XU Ming-yue, XIAO Qing-sheng, ZHANG Xue-kun, CHENG Yong, LU Guang-yuan, XU Jing-song, ZENG Liu, FU Gui-ping, QU Zhen, ZOU Xi-ling*
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    Correlation between gene expressions and physiologies was investigated under drought stress using 8 tolerant and susceptive rapeseed materials. The genes included 6 drought-related genes (btg-26, BnD22, P5CS, CaM, BADH and ADC). Results showed that the expression profiles were different among the genes between tolerant rape Q2 and susceptive materials Zs6 and S95-3. Among the 5 indicators, leaf wilting index (LWI) and relative electrolyte leakage (REL) varied largely among the varieties under drought. Correlation analysis showed that after 3h of drought, P5CS was significantly correlated to LWI and REL, BADH was correlated with REL. After 24h of drought, btg-26 was significantly correlated with relative water content and REL. Results indicated that the above genes could be appraisal indices of drought-tolerance. 

     

  • YANG Wan-ming, WANG Min, DU Wei-Jun*
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    A backcross inbred line population was developed from a crossing between SNWS0048, a wild soybean and cultivar Jinda73. Traits related to seed germination of this population was examined under PEC stress, major indexes and drought tolerance of soybean seedlings were analyzed by principal component analysis method and membership function method. The result showed that greater germination index of drought tolerance,higher relative germinability, relative germination rate and related germination index were preferred characteristics of drought-tolerant varieties under PEG stress. Water absorption and root traits were indirect indexes of evaluating drought-tolerance. The analysis of principal component and comprehensive subordinate function showed that 200 lines were divided into different drought tolerant levels. Out of them, 10 drought tolerant lines were screened out. 

  • ZHANG Peng,XU Chen,XU KE-zhang*,ZHANG Zhi-an,LI Da-yong,JI Ping,FENG Yin-di
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    To search the simple identification method of salt tolerance in soybean cultivars and understand soybean cultivars response to salt tolerance at different stages, 80mmol/L salt solution was used to identify salt tolerance in soybean cultivars from seed germination. The results showed that 72h germination percentages with 80mmol/L salt solution soaking could be used as a method in salt resistance identification and screening. The root length, number of fibrous root, dry weight and fresh weight of roots were closely correlated with the salt tolerance after seed germination. Soybean cultivars were the most sensitive to salt stress at seeding stage. The salt-sensitive cultivars cannot grow seedlings with 80mmol/L salt solution and this period is also important for understanding salt tolerance of soybean cultivars. At vegetative stage (V2 and V4), the height and biomass of plant were decreased significantly under salt stress. Salt tolerances were the greatest and biomasses had the least decrease at R2 and R4 stages. Salt stress showed the greatest damage to the plant at R6 stage, namely leaf abscission and premature aging. The salt tolerance of germination period was consistent with that of late growth of plants. Germination percentage of germination period can be used to identify salt tolerance of plants.

  • CAO Xiao-ning, SUN Shi**, WU Cun-xiang, HAN Tian-fu*, YANG Wen-yu*
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    Effects of different cut-in grafting methods on the survival rate and recover day of grafted soybean plants in soybean young seedlings (Vc-V1) stage were studied with the aim to develop a grafting technique to study the shoot-root relationship of soybean. The cut-in grafting method was used in the study, and two treatments were designed as follows: scion with and without cotyledons were grafted on the stock with cotyledons; scion with cotyledons were grafted on the stock with and without cotyledons, respectively.The results showed that the mean survival rate of cut-in grafting method was more than 90%. The recover days of stock with cotyledons and scion without cotyledons treatments were 3-4d. The treatment remained scion cotyledons and stock cotyledons showed a significant impact on the survival rates of the grafted plants. Scion treatments did not show significant impact on the survival rates of stock with cotyledons treatment. The difference of grafting site did not affect the survival rate of grafted plants. The combination of stock with cotyledons and scion without cotyledons treatments was superior to others. 

  • WEN Jing, ZHAO Gang, JIAO DAN, ZHAO Gang,WANG Jiang-hua, Hu Bao-zhong*
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    Dongnong 42 and its dwarf mutant-HK808 were used to analyze the external morphochoresis and its internal anatomical structure in different growing stage. The results indicated that the difference of height between the two test materials is determined by the different length of the internode 5-14. The anatomical structure analysis of root indicated that the Dongnong42 was much greater than HK808 in vascular area, there was a complementary relationship between the dynamic curve of the vascular area and the curve of vascular number in different varieties. Dongnong42 was significantly lower than HK808 in the bark/root ratio; there was significant difference in the area and number of vascular and parenchyma cells in stem between the two varieties, there is no significant difference in their vascular number. The anatomical indexes between the two varieties in petiole had the similar change pattern to that in stem. Leaf thickness and area of vascular of Dongnong42 was significantly higher than HK808 in leaf, palisade/spongy of HK808 is significantly higher than Dongnong42. There were significant differences between the two tested materials in the size of chloroplast, the number of chloroplast grana lamellar and the morphology of grana lamellar.

  • CHEN Ye,WAN Li-li,WANG Zhuan-rong,HONG Deng-feng,YANG Guang-sheng*
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    The disease-resistant gene Ospgip1 from rice was introduced to the B. napus materials 7-5, P61-5 and T45 by Agrobacterium-mediated transformation respectively and the positive plants were obtained. T1 transgenic plants of Ospgip1 were confirmed by PCR, RT-PCR and Real-time PCR. The results showed that Ospgip1 were consistently inherited and expressed. The Sclerotinia inoculation test showed transgenic plants were disease-resistant, and it showed significant negative coefficient between the Ospgip1 expression and the lesion size at the seedling stage. The lesion average size of 16 transgenic lines was smaller than the control at the seedling stage, but 21 transgenic lines were smaller than the control at the adult stage.

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  • GUO Xiao-jing,GONG Yang-min, LIANG Zhuo, WAN Xia, HU Chuan-jiong, LIU Jiao, JIANG Mu-lan*
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    Culture conditions were adapted to obtain larger biomass and higher docosahexaenoic acid (DHA) yield of Crypthecodinium cohnii under flask fermentation level. Results showed that the optimal culture should be at 22℃, in carbon-source culture with 12g/L glucose for 4d and then shifted to the same media but with 3g/L sodium acetate instead for 4d. Under this optimal condition, the C. cohnii biomass yield, DHA yield and DHA content reached 234.85mg/(L•d), 17.45mg/(L•d), 74.36mg/g dry weight respectively.

     

  • WANG Li-jun, WANG Hong,XIE Le, FU Xiao-hua, LIU Hai-bin,ZHANG Lan-tian
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    Qualitative and quantitative analyses for palm oil in pure peanut oil were completed by raman spectroscopy. The figure of 2-dimension factor by factor analysis could make a distinction between the mixed oil and pure peanut oil, and the established linear equation by the method of least squares was used for the quantitative analysis.

  • WU Jian, TU Jiang-ying, ZHOU Yong-ming
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    Sclerotinia stem rot of oilseed rape (Brassica napus) caused by fungal pathogen Sclerotinia sclerotiorum is one of the most devastating diseases worldwide. It is one of the most important biotic stressors affecting oilseed rape production. Breeding and cultivation of resistant varieties is the most efficient, economic and environmental friendly approach to control the disease. In this paper, research progress was reviewed in aspects of S. sclerotiorum pathogenesis, rapeseed resistance heredity, gene expression profiles responding to the disease infection and application of resistance genes in oilseed rape.