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  • 2015 Volume 37 Issue 1
    Published: 28 February 2015
      

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  • CHU Pu,ZHAI Li-na,GUAN Rong-zhan*
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    Sterol carrier protein 2 (SCP-2) is a small, basic protein involved in intracellular lipid circulation, fatty acid metabolism and peroxisomal β-oxidation. Based on the homologous sequence, a cDNA fragment of SCP-2 gene was cloned in Brassica napus, and designated as BnSCP-2. The complete open reading frame of BnSCP-2 is 372bp in length, encoding 123 amino acid with a predicted molecular mass of 13.52kD and a pI of 9.35. The amino acids of SCP-2 protein were highly conserved. Phylogenetic analysis revealed that deduced BnSCP-2 protein was close to SCP-2 protein in Arabidopsis. Realtime RT-PCR showed that BnSCP-2 was expressed in all tissues tested, with the highest expression in flowers and flower buds. Its expression in seedlings was significantly up-regulated by GA3, 6-BA and ABA treatments, suggesting that BnSCP-2 may be involved in growth regulation and young seedling development.

  • ZHANG Feng-qi, CHENG Xiao-hui, LIU Yue-ying, TONG Chao-bo, DONG Cai-hua, YU Jing-yin, HUANG Jun-yan*, LIU Sheng-yi
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     Branch number is an important trait affecting yield of oilseed rape. To improve breeding efficiency on branch number improvement, QTLs on branch number were identified by using single environment and multi-environment detection methods. The mapping was conducted in a recombinant inbred lines (RILs) population derived from 888-5 (with more branch number) and M083 (with less branch number) by employing newly developed Brassica napus 60K SNP array. Results indicated that a total of 18 putative QTLs in 4 environments (Wuhan-2012, Wuhan-2013, Yangzhou-2012, and Ynagzhou-2013) were found on chromosome A2, A6, A7, C1 and C4. Among them, 11 QTLs were repeatedly detected in more than 2 environments, and 2 QTLs showed interaction with environments. 2 major effect QTLs (qBN2-3 and qBNE2-1) were repeatedly detected in 3 and 4 environments respectively, explaining 13.12%-20.60% and 2.80%-30.10% of phenotypic variations. QTL qBNE2-1 presented interaction with environment. By comparative mapping of the SNPs flank sequences with B. napus genome, 4 candidate genes were found from genomic regions (279kb, 165kb and 562kb) of 3 QTLs (qBN2-1, qBN7-6, and qBN7-8 which explained 19.40%-17.30%, 5.70%-12.21%, and 7.88%-10.32% of phenotypic variations respectively). Their orthologous genes in A. thaliana were CUC2, PIN3, F23N20.8 and PIN4, all involving in branch number differentiation in A. thaliana. 

  • FANG Yan,YANG Gang, SUN Wan-cang*, WANG Li-ping, ZHANG Shu-juan, YANG Jian-sheng, LIU Lin-bo, LIU Zi-gang, ZENG Xiu-cun, WU Jun-yan
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    To evaluate germplasm for winter turnip rape (Brassica rapa L.) breeding on cold resistance in north China, genetic diversity of 19 turnip rape varieties (which were significantly different in temperature sensitivity, cold hardiness and oil quality) were analyzed using 9 SSR and 36 InDel markers selected from 252 markers. Results showed that 95 allelic variations were detected by 45 SSRs and InDels, with an average of 2.15 alleles each marker. Effective alleles varied from 1.05 to 3.27, with an average of 1.70 each. Shannon index ranged from 0.121 7 to 1.269 5, with an average of 0.580 3. Polymorphism information content (PIC) ranged from 0.049 9 to 0.049 9, with an average of 0.308 1. By NTSYS calculation and unweighted pair-group method with arithmetic mean (UPGMA), genetic similarity (GS) of 19 materials varied between 0.52 to 0.86. They were divided into 2 major groups (I-1 and I-2) with multiple ventricular at GS 0.605. Group I-1 was divided into 2 semi-groups (II-1 and II-2) according to their winterness at GS level of 0.655. Among them, winter-type rapes were divided into 4 small groups at GS 0.71. The cluster results showed abundant genetic diversity with large distance between varieties from different geographic origins and cold hardiness in north China.

  • GAO You-fei, YUE Yan-lei, JIANG Bing-jun, HAN Tian-fu
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    GmFT2a is a key gene in photoperiodic pathway of soybean. However, the underlying molecular mechanism of GmFT2a expression is not clear. In this study, a photoperiod-sensitive soybean cultivar Zigongdongdou and insensitive cultivar Heihe 27 were used as materials to compare the GmFT2a promoter sequences. It revealed that there were 2 insertion/deletion (InDel) fragments, T1 and T2, which contained light reaction elements in the GmFT2a promoter of Zigongdongdou. However, the same fragments were not in that of Heihe 27. The bait constructs were prepared according to the sequences of light reaction elements of I-box and SP1 to screen cDNA library of Heihe 27 leaves, and a MYB transcription factor bound with T1 was isolated with the yeast one-hybrid system. The obtaining of the MYB transcription factor provided a favorable clue for further study of the molecular mechanism GmFT2a expression regulation. 

  • GU Xiao-na, LIU Zhen-ku, WANG Pi-wu*, ZHANG Zhuo, MA Jian, QU Jing, ZHANG Jun,FU Yong-ping, LU Shi, ZHENG Cheng-zhong, LIU Ting-ting
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     T5 generation of transgenic soybean-lines JN29-705-15 and JL30-187 with hrpZPsta gene were investigated by real-time quantitative polymerase chain reaction technique (qRT-PCR) to analysis the relationship between expression of the target gene in different tissues of the transgenic soybean and their resistance to Phytophthora root rot and gray leaf spot. The results showed hrpZPsta gene was expressed in soybean leaves, stems, roots and seeds, the average relative expression levels of the two strains were 8.2/6.1, 0.9/0.7, 6.5/4.6 and 0.8/0.7, respectively; the resistance to Phytophthora root rot and gray leaf spot disease were improved compared with wild-type; compared to control, the resistance against Phytophthora root rot disease of JN29-705-15 increased from sensitive to medium-resistant; and resistance of JL30-187 increased from medium to full-resistant; the expression levels of hrpZPsta gene in leaves was positively related to resistance of gray leaf spot, and significant negative correlation to the disease levels. These results indicated that the expression level of hrpZPsta was correlated to resistance to both Phyophthora root rot and gray leaf spot in transgenic soybean.

  • WU Lie-hong, LI Fu-zhen, WU Xue-long, SONG Du-lin, WANG Mei-xing
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     The content of α-linolenic acid varied in different germplasm resources and different seed developmental stages in peanut oil. Oneα-linolenic acid synthesis related gene ω-3⊿15 fatty acid dehydrogenase gene, AhFAD3A was cloned from cotyledonary tissues of peanut germinating seeds using RACE and bioinformatics analysis. Its expression patterns were analyzed in different tissues and different period of seed developmental stages by the fluorescence quantitative real-time PCR method between two α- linolenic acid synthesis genes, AhFAD3A and AhFAD8. The results showed that: AhFAD3A gene was mainly expressed in leaf, root and young seed tissue during the period of flowering, and relatively low expression was found in the other developmental stages. This result verified that the gene expression was positively correlated to the formation of α-linolenic acid in seed. AhFAD8A gene was expressed in the green tissue during the whole growth stage of peanut, not detected in non optical tissues. These results laid a foundation for further research in determining the function of AhFAD3A gene, its expression regulation and its relationship with the formation ofα-linolenic acid in seed. 

  • XUE Jian-feng, TAN Mei-lian§, YAN Ming-fang, WANG Lei, FU Chun-ling, WANG Li-jun, YAN Xing-chu*
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     To investigate the genetic diversity and phylogenetic relationship of castor, genetic diversities and relationships among 30 domestic cultivars and 1 French cultivar were assessed by SSR and EST-SSR markers. Results showed a moderate diversity level among these cultivars, with average 2.267 alleles per locus, 0.553 Shannon index, 0.345 Nei's expected heterozygosity and 0.289 polymorphism information content (PIC) respectively. The 31 castor cultivars were clustered into 7 groups at the similarity coefficient of 0.730. Most cultivars were clustered into the same breeding units or regions. Among the 171 primers, 10 were used to construct a clear and polymorphic footprint,and can be used in the identification of cultivars.

  • TAO Xia, LI Hui-lin, WAN Lin, ZHOU Qin, JIANG Hai-dong*
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    To understand the alleviation effect of indole-3-acetic acid (IAA) on waterlogging stressed rapeseed at budding stage, pot experiment was carried out to investigate the growth and physiology of stressed Brassica napus cv Qingyou 7 (susceptive) and Suyou 4 (tolerant) plants before and after IAA (0.1 mmol/L) foliar spray. Results showed that 10 days of waterlogging inhibited rapeseed growth, biomass and chlorophyll content decreased. The increased physiological index included enzyme activities of SOD (superoxide dismutase), sucrose synthase, sucrose phosphate synthase, contents of GSH (glutathione), AsA (acrylonitrile styrene acrylate copolymer), MDA (malondialdehyde) and soluble sugar. When IAA sprayed after waterlogging, the alleviation effects were significant, especially on susceptive cultivar Qingyou 7. The leaf photosynthetic pigment contents were increased, which started promoting plant growth 5d after spray. SPS (sucrose phosphate synthase) activity decreased, resulting reduced sucrose. Decreased SS (sucrose synthase) activity reduced sucrose decomposition. After 15d of spray, root dry weight of both cultivars increased significantly, and stem dry weight increased in the susceptive. Yield traits recovered significantly, especially on pods per plant and seeds per pod. But the effect of IAA after waterlogging was less effective on oxidation system. 

  • YUAN Xiao-yan, FU Ming-lian*, LI Gen-ze, WANG Jing-qiao, WEI Sheng-guang, CHENG De-rong
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    To evaluate new drought resistant germplasm, 24 DH lines from hybrid of Brassica napus and Brassica juncea were planted under 2 conditions of drought stress at 3 stages (seedling, flowering and podding stages) and normal irrigation in Kunming, Yunnan Province. In this area, the rainfall is approximately 60mm in the whole growth season for rapeseed. The identification criteria for drought-resistance were yield, 4 yield-related agronomic traits and five quality traits and different evaluation methods were compared by drought resistance index method, gray relevancy method and cluster analysis. The result showed that the drought resistance of the most hybrids was stronger than that of the conventional B. napus, especially at podding and seedling stage. 10 strong drought-resistant materials were screened, and 2 materials with both strong resistance and high yield were found. The drought-resistance correlation coefficient analysis of 3 growth stages showed that the drought resistance at seedling stage was correlated to flowering stage, and podding stage showed a poor correlation to seedling or flowering stage. Drought stress at any growth stage could seriously affect the fresh weight, plant height, oleic acid content and erucic acid content. Yield was affected by the drought stress most seriously at the flowering stage, and next at the seedling stage. The drought-resistance had significant positive correlation to fresh weight at seedling, flowering and podding stages. The drought-resistance had significant positive correlation to plant height at flowering stage.

  • YAN Yan-hong, CHEN Zhong-qun, WANG Xiao-chun, YONG Tai-wen, LIU Wei-guo, YANG Wen-yu*
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    The effect of molybdenum (Mo) fertilizer on dry matter accumulation, distribution and yield of relay strip intercropping soybean were studied by completely randomized block design. The results showed that dry matter accumulation of the whole plant increased at first and then decreased with the increase of Mo application amount from 5 internodes stage (V5) to full mature stage (R8), and it was the highest under B2 (1.0 g/kg) treatment. The dry matter was mainly distributed to stem and leaves, and the distribution proportion of the dry matter was decreased to stems and increased to leaves, with Mo application at V5 stage and full-bloom stage (R2). The distribution proportion of the dry matter to legumes was the highest, and it increased initially and then reduced with the increase of Mo application. But the changes of the distribution rate of the dry matter to stems and leaves with the increase of Mo application were all opposite to that of legume. The dry matter translocation and contribution ratio from vegetative organs to legumes and yield were increased with the Mo application, and the effect of B2 treatment was the best.

  • DENG Jun-Cai, LIU Jiang, LEI Ting, YANG Feng, SU Ben-Ying, CUI Liang, WAN Yan, HUANG Shan, YANG Wen-Yu*
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    To investigate the effect of seed mildew during harvest season on soybean yield and quality, 18 soybean varieties were treated by using the artificial rainfall chamber to simulate the continuous rainfall weather. The results showed that seed mildew caused great yield loss, the loss rate ranged from 23.14% to 96.55%, and the loss rate was highly significantly related to mildew index (P<0.01), the regression equation between yield loss rate(Y) and mildew index(X) was Y=1.34X+24.51,R=0.98. The seed mildew also induced soybean seed quality deterioration. The relative content of protein and fat reduced firstly and then increased with the increased seed mildew grade, the relative content of soluble sugar decreased with the increased mildew. The absolute content of the three types of nutrient substances decreased with the increased seed mildew grade, and the 100-seed weight decreased significantly. The results also showed that the resistance of black soybean and brown soybean to field seed mildew was stronger than that of yellow soybean,indicating that soybean coat color had some special relationship with the resistance to seed mildew in field. The seed mildew caused by continuous rainfall weather during harvest had detrimental effects on soybean yield and quality, while the significant differences in sensitivities among soybean varieties benefited the breeding of the cultivars with high resistance to seed mildew.

  • ZHU Xiao-feng, LI Dong-hua, WANG Lin-hai, ZHANG Yan-xin, ZHANG Xiu-rong*
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     The contents of 3 endogenous phytohormones including indole acetic acid (IAA), gibberellic acid (GA3) and abscisic acid (ABA) in the 3rd leaves from top and stem tips of two dwarfs and two high genotypes were investigated by liquid chromatography-tandem mass spectrometry, in order to analyze their dynamic changes and their relationship to growth rate. The results showed that these 2 genotypes kept growing from the 3-true-leaf stage to terminal flowering stage, but the daily growth rate of the high genotypes was greater than that of the dwarf genotypes. During various developmental stages of plant height in 2 types of material, content variation of IAA in leaf and GA3 in stem tip were observed. There was no clear change pattern of ABA content between 2 types of sesame. Compared with other 3 genotypes, the ABA content in ZZM2748 was significantly higher on 20th day post anthesis (DPA) both in leaf and stem tip and was similar to others during other growth period, indicating that there might be a mechanism that strongly inhibited stem growth. Correlations analysis showed GA3 and stem growth of sesame had the strongest correlation.

  • SUN Chao-chao, REN Li, CHEN Kun-rong, XU Li, LIU Fan, FANG Xiao-ping*
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     To understand the risk of rapeseed clubroot in different soil environments, 12 soil-samples were collected from main rapeseed production areas in 2012 to 2013. Plasmodiophora brassicae were inoculated on high susceptive Brassica rapa cv Beijingxin 3 for club root risk assessment. Results showed a significant relation between disease incidences and sampling location. The disease incidences were up to 76.7% on samples from Suizhou of Hubei Province and Chaohu of Anhui Province under the resting spore concentration of 103 spores per soil gram. On the contrary, the disease incidences were as low as 0-3.3% in soils from Wuchang and Yangluo of Hubei Province, Xining and Huzhu of Qinghai Province, Minle of Gansu Province, even under the inoculation density of 105 spores per soil gram. It suggested that clubroot risks were high in the soil from Suizhou and Chaohu, and low from Wuchang, Yangluo, Xining, Huzhunghai and Minle. Some soil factors were found inhibiting the disease after inoculation tests on sterilized soil samples. The correlation of environmental factors in soils and clubroot incidence were determined by path analysis using the statistical analysis software SPSS (statistical product and service solutions). Results showed that pH, available calcium, total phosphorus, total potassium and cation exchange capacity (CEC) were negatively related with clubroot incidence. These results could benefit the development of clubroot management strategies in effective prevention of clubroot disease.

  • WANG Xue, DUAN Yu-xi*, CHEN Li-jie, WANG Hui, WANG Yuan-yuan, ZHU Xiao-feng
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    The objective of this study was to set up the differentially expressed map for soybean root proteins induced by Heterodera glycines (SCN), and to explore the resistant mechanism of the pathogen at molecular level. The cross was made between resistant cultivar Harbin Xiaoheidou (ZDD7170) and susceptible cultivar Liaodou 10, the proteomic differentially expressed of soybean roots induced by SCN was analyzed. The results showed that there were difference between proteins extracted from resistance pool sample and sensitive pool sample. About 400 protein spots were detected, including 3 more than 3-fold increased points and 9 more than 3-fold decreased points in resistance pool sample, 6 specific proteins expressed in resistance pool sample and 10 specific proteins expressed in sensitive pool sample. 16 protein spots were obtained by Mascot database searching and inter-comparison preliminary identification, all of those proteins as part of the resistant network or development related proteins were related to plant defense responses, energy metabolism, cell signal transduction and transcriptional regulation.

  • KANG Yan-ping, YAN Li-ying, LEI Yong, LIAO Bo-shou*
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     Peanut virus diseases can cause mosaic and dwarf symptoms on peanut plants and influence peanut’s production. Reverse transcription PCR(RT-PCR)was used to detect peanut viruses in 12 peanut samples collected from Gaocheng, Hebei Province. Results revealed that only Peanut stripe virus (PStV) was detected in the 90% of the collected samples with mosaic and dwarf symptom. The coat protein gene (cp) of PStV of two isolates were sequenced, and their sizes were 864 base pairs. Sequence analysis showed that the sequences of Gaocheng isolates shared 94.4%-99.2% and 94.7%-99.7% similarities at nucleotide level and 95.5%-99.7% and 95.8%-100.0% at amino acid level with those of other PStV strains. These isolates shared closer relationship with Chinese and USA isolates compared with Chinese Taiwan, Thailand and Vietnam isolates by phylogenetic analysis of CP nucleotide sequence.

  • LIU Si-lei, LIU Wei, DONG Xu-yan*, WEI Fang, WANG Xiang, LV Xin, ZHONG Juan, WU Lin, CHEN Hong*
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    A lipase catalyzed synthesis of 1,3-diacylglycerols with microbial oil rich in ARA was established. The effects of reaction time, reaction temperature, substrate molar ratio and lipase dosage on 1,3-diacylglycerols yield were systematically investigated. Based on the single factor experiments, response surface methodology (RSM) was used in optimization of technological parameters. The optimal conditions were: reaction time 2.7 h, reaction temperature 57℃, substrate molar ratio 2.5:1, lipase dosage 7.9%. Under the optimal conditions, 1,3-diacylglycerols relative percentage was up to 73.5% and 1,3-ARA-DAG content was 38.1%.

  • YANG Wan-feng, LIU Yan, LIU Xiang, SHAO Pei-ze, CHEN Yun-qing, ZHAO Wen-jun
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    order to quickly test Pseudomonas savastanoi pv. phaseolicola in imported soybean, primers 52B/8F and 24B/24F were designed based on argK gene of P. savastanoi pv. phaseolicola. We established nested PCR and tested its specificity and sensitivity. Specificity assays showed that only the isolates of P. savastanoi pv. phaseolicola were positive, and the other tested pathogens were negative. The sensitivity of detecting DNA and bacterial suspension reached 0.916×10-4 ng/μL and 2.4×103 CFU/mL respectively. The sensitivity of nested PCR assay was 1 000 times higher than normal PCR. 3 contaminated samples among 100 imported soybean were detected and the specific DNA band was obtained by established nested PCR, and the sequenced results showed that these 3 soybean samples were indeed infected by P. savastanoi pv. phaseolicola. The nested PCR developed in this study had properties of specificity, high sensitivity, and can be rapidly and easily operated. It could be used in detecting P. savastanoi pv. phaseolicola for imported soybean quarantine.

  • XU Lin,ZHANG Zhao-wei,LI Pei-wu*,ZHANG Qi*,ZHANG Wen
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    To develop quantum dot application on food safety detection, CQDs (carbon quantum dots) and GQDs (graphene quantum dots) were evaluated as the fluorescence labels in aflatoxins detection in cereals and oils. Using environment friendly method, GQDs were synthesized directly in citric acid solution, and showed the strongest fluorescence intensity under 0.375g/mL citric acid. Then the GQDs and CQDs were covalent coupled with monoclonal antibody 1C11 by 1-ethyl-3- (3-dimethylaminoproyl) carbodiimide hydrochloride (EDC) and N-hydroxyl-sulfosuccinimide sodium salt (NHS). Results showed that both quantum dots had strong fluorescence intensities, which could be served as fluorescence labels in aflatoxin immunoassay. CQDs fluorescence property was stronger than that of GQDs after coupled with antibody.

  • LUO Peng, FENG Yong-kang
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    Researchers in Laboratory of Plant Genetics, Sichuan University, first reported the production of induced parthenogenetic haploids in rapeseed in 1976. In the next 20 years since then, the authors and coworkers carried on systematic studies on biological mechanism and genetic improvement of induced parthenogenetic haploids. They obtained higher-yielding hybrid combinations, such as 3529×Oro and 3529×Swiss in B. napus. In this brief review, systematic studies of induced parthenogenetic haploids in B. napus were discussed, and further studies were suggested.

     

     

  • GUO Yan, ZHAN Gao-miao, YANG Xiao-yan, HUA wei, LV Ying-tang*
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     A leaf-specific expressed gene ACD6 and 2 silique-specifically expressed genes (AT2FP2, at1g56100) of Arabidopsis thaliana were obtained from Genevestigator by bioinformatics search. Gene expression patterns were detected employing real-time quantitative PCR. The upstream regions of these genes were obtained from TAIR website, and promoter sequences were predicted by online software PlantCARE and BioProspector. The promoters were named as Pat4g14400, Pat5g57520 and Pat1g56100 respectively. In corresponding transgenic A. thaliana plants, histochemical GUS staining showed that ACD6 was mainly expressed in leaf, Pat4g14400 was leaf-specific. AT2FP2 and at1g56100 expression in silique were both several times higher than in other tissues. It could be concluded that Pat5g57520 and Pat1g56100 were silique-specific.