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  • 2013 Volume 35 Issue 1
    Published: 28 February 2013
      

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  • WANG Zhi-hui, TONG Chao-bo, YUAN Wu-zhou, LIU Xue-qun, CHENG Xiao-hui, YU Jing-yin, DONG Cai-hua, LIU Sheng-yi
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     Bacterial-type PEPC genes were cloned from four oil crops (rape, soybean, peanut and sesame). Their sequences of conserved domains (BOX I to BOX IV) and PEPC activity sites were characterized. The genes included Bna10093361, Bna1009749 and Bna10093360 from Brassica napus, Glyma10g34970.1, Glyma01g22840.1 and Glyma02g14500.1 from Glycine max, SIN1018296 from Sesamum indicum, AhPPC5 from Arachis hypogaea. The 8 genes were commonly composed of 19 to 21 introns, and contained one unique and highly divergent insertion of about 356 to 600bp. PEPC sequences had R/KNTG in C-terminal tetrapeptide, but lacked the phosphorylation motif at N terminus. Results from RT-qPCR showed that Bna10093360 expressed with less than 0.1% of rapeseed BnaActin, Glyma10g34970.1 and Glyma02g14500.1 expressed with less than 2% of soybean GlymaActin. AhPPC5 expressed 32% to 175% of peanut AhActin, and SIN1018296 expressed 3% to 18% of sesame SINActin. The expression trends of AhPPC5 and SIN1018296 were higher at early seed developing stage and then decreased. Results indicated that the bacterial-type PEPC gene functions were widely differentiated.

  • PENG Peng-fei, HU Qiong, LI Yun-chang, FU Li, CHEN Yu-feng, LIU Jia, MEI De-sheng
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    The cDNA fragment of a putative pod shattering gene BnRPL was cloned from Brassica napus line R2 (easy to disperse seed) by RACE according to conserved region of AtRPL gene. Results showed that two BnRPLs were amplified named as BnRPL.a [with an open reading frame (ORF) of 1 758bp, coding 585 amino acids] and BnRPL.c (with an ORF of 1 734bp, coding 577 amino acids). Each BnRPLs included 3 introns and 4 exons. BnRPL.a and BnRPL.c was 92% identical. Comparing to RPL sequences from other species, BnRPL showed high homologous to AtRPL from Arabidopsis thaliana and RPL from Lepidium campestre. Variation of RPL protein occurred during the divergence from dicots and monocots. RT-PCR expression analysis showed that BnRPL gene was highly expressed in developing siliques. Moreover, it expressed in maturing silique wall and pseudoseptum, but not in seeds.

  • LI Yang,GU Hui,QI Cun-kou
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    To identify the functions of lodging resistance related genes from Brassica napus L., the expression patterns of lignin biosynthesis key genes (F5H, 4CL and COMT) were investigated and compared between cultivar Zheping 1 (lodging-resistant) and Gaojie 1 ( lodging susceptible). Quantitative real time PCR was used to analyze the gene expressions in root collar and stem at bolting and silique-setting periods. Results showed that, for Gaojie 1, the relative expression level of F5H was higher in root collar, and lower in stem. The expressions of 4CL and COMT of Gaojie 1 were much higher in root collar and stem at bolting stage and in root collar at silique-setting stage. But they were significantly (P<0.05) lower in stems at silique-setting stage. It can be concluded that higher relative expression of F5H, 4CL and COMT in stems at silique-setting stage would be beneficial in enhancing lodging resistance for B. napus.

  • LAN Lan,WU Shuai, SHEN Li-wei, WANG Zhi-kun, MENG Fan-li, SONG Bo, TUO Yun,
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     The transgenic plants were obtained by Agrobacterium-mediated transformation of soybean (DongNong 50) cotyledon nodes. The transformation of cryIA was confirmed by PCR,RT-PCR and southern blot analysis. The resistance to soybean pod borer of transgenic plants was detected by bioassay method. 114 PPT-resistant plants were obtained and 4, 3and 9 positive plants from T0, T1 and T2 generations respectively were tested by PCR. 1800bp purpose fragments were detected by RT-PCR analysis in 3 T1 transgenic plants and the hybridization bands could be detected by southern-blot assay in two T1 transgenic plants. The gene of cryIA was successfully integrated into the genome of transgenic plants and could be transcripted normally in T1 generation. Additionally, the results of bioassay indicated that the transgenic plants displayed high insect-resistance to soybean pod borer. Further investigations on the genetic stability of cryIA in different transgenic lines and generations are to be carried out.

     

  • ZOU Zhi
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    Castor bean (Ricinus communis L.), a tropical shrub that belongs to the Euphorbiaceae family, is an important non-edible oilseed crop widely cultivated in tropical, sub-tropical and temperate countries for its high economic value. Although it’s well known for its plasticity to unfavorable environments, little is known about the molecular mechanism of castor bean plant tolerance to stress to date. The genome sequence released lately provides valuable resources to address this issue. In this study, with the published genome and EST sequences, a genome-wide search was carried out for genes encoding RcWRKYs, a type of plant-specific transcription factor which was shown to be one of the most important transcription factor families involved in biotic and abiotic stress response of plants. As a result, a total of 56 RcWRKYs were identified and then nominated as RcWRKY1-56 based on their gene structure and phylogenesis. The RcWRKYs containing one to five introns each were distributed across 39 of the 25,800 scaffolds. According to the number of WRKY domain and the type of zine finger, the identified RcWRKYs were grouped into I, IIa, IIb, IIc, IId, IIe or III class or subclass. Comparative genome analysis suggests that the Ricinus WRKY family didn’t expand significantly in the history for most of them have ortholog genes in Arabidopsis thaliana. Among different class or subclass of RcWRKYs, this class may have evolved in a faster pace.

  • KE Tao, MAO Han, DONG Cai-hua, CHEN Shu-yuan, YU Jing-yin,ZHAO Ying-zhong,LIU Hong-yan,LIU Sheng-yi
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        Currently, the lack of EST sequence information has limited the discovery of sesame EST-SSR primer number. All the 45569 EST sequences of high oil content sesame species Zhongzhi14 were obtained from a full-length developments seeds cDNA library and assembled 32421 unigenes. The unigenes represented about 17.428 Mb of genic sequences. Using MISA and SSR finder software to screening SSRs, 1949 non-redundant (NR) SSRs were identified in 1688 uniESTs. A total of 226 sequences contained more than one SSR. The trinucleotide repeats are the dominant types with the frequency of 74%. All of the designed 34 SSR primer pairs related to oil synthesis genes could amplify characteristic SSR fragments against 12 different oil content species, and 3 primer pairs amplified polymorphic fragments. The EST-SSR markers of sesame are highly efficiency, and should be the ideal tools for studies of genetic variation, genetic diversity, and development of genetic linkage maps of sesame.

  • HU Cheng-wei, ZHANG Xue-kun, ZOU Xi-ling, CHENG Yong, ZENG Liu, LU Guang-yuan

    A new method based on sand culture and 14% (m/V) polyethylene glycol-6000 (PEG-6000) was developed to evaluate root traits of 39 rapeseed genotypes at seedling stage and the connections between root traits and drought tolerance. Results showed that root weight and lateral-root number were significantly inhibited by drought, while root length and root-to-shoot ratio were modestly affected. Significant correlations were found between vigor index and both lateral roots number and root length, implying that these two traits can be used as selecting criteria for drought tolerance. Inbred lines and hybrid varieties had no significant difference on root traits for drought tolerance. By clustering analysis, the 39 genotypes were divided into 5 groups at Euclidean distance of 60. Among them, group A was considered as the most tolerant to drought, and group E was the most sensitive. Three drought tolerant genotypes (OR918, OR805 and OR2025) showed potentials for further breeding program.

  • WANG Xiao-guang, JIANG Chun-ji, ZHAO Xin-hua, YU Hai-qiu, CAO Min-jian

    Five high yield peanuts varieties, Xuhua 5, R03-3, Tangyou 4, Fenghua 5 and Shanyou 101 were selected from 109 varieties introduced from other regions of China, and their flowering characteristics in Liaoning Province were compared. Results showed that five varieties began to flower on the 27th to 36th days after seedling. Tangyou 4 flowered earliest,duration of flowering was longest (50d), total flower number (258) and effective flower (87) number per plant were higher. In comparison, Xuhua 5 and R03-3 (June 28) flowered late, duration of flowering (37 days for Xuhua 5 and 41 days for R03-3) was short, and the total number of flowers per plant (160 and 185 respectively) was small. Fenghua 5 and Shanyou 101 were intermediate in these traits. The total number of flowers on the first pair and second pair of lateral branches of Xuhua 5 accounted for 86.8% of the total flower number of the plant, representing the highest proportion among those varieties. The proportion of Shanyou101 was the lowest (74.9%). There were more flowers at the first and second nodes of the lateral branches than in other nodes, where only 2~4 flowers were detected. The flowering dynamics per 5 day for Xuhua 5, R03-3 and Fenghua 5 exhibited the single-peak curves, while the flowering dynamics for Tangyou 4 and Shanyou 101 showed bimodal curves. For all the 5 peanut varieties, after peak period, the number of flowers per plant within 5 days decreased rapidly.

  • DENG Xiao-dong,WU Xiao-xia, FAN Xin-zhao,FEI Xiao-wen,REN Da-ming
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     A high oil microalgae strain collected from fresh waters of Beibu Bay area was indentified as Chlorella sp. and cultured in high-carbon medium TAP. Effects of IAA and ABA on its biomass and oil accumulation were studied. The results showed that 10μmol/L IAA promoted the growth of algae strains significantly. The max biomass increased by 58.24% compared to control while oil content decreased by 34.71%. In contrast, 20μmol/L IAA clearly inhibited the growth of algae strains and the decrease in max biomass was 53.60%. However, oil content increased by 95.46%. The inhibition of ABA on the growth of algae strains was evident. There was a 57.52% decrease in its biomass compared to control group and a 121.49% increase in its oil content. The protein, total sugar content, chlorophyll content and photosynthetic efficiency of the algae were decreased compared to control when IAA(20μmol/L)and ABA (2μmol/L)were added to the medium.

  • XIE Li-hua, YU Chang-bing, HU Xiao-Jia, LI Yin-shui, LIAO Xing, ZHANG Shu-jie, HE Hui, CHENG Yu-gui, ZHANG Zong-ji,LIAO Xiang-sheng, CHE Zhi
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    To improve the fertilizer efficiency in oilseed rape under direct seeding and high density planting (as 375 000 plant per hectare), a “3414” design were used to study the optimum rates of nitrogen (N), phosphorus (P) and potassium (K) fertilizers in oilseed rape (Brassica napus L.) in field. Experiments were carried out in three sites in Guilin of Guangxi Province, Zhijiang of Hubei Province and Yulong of Yunnan Province. Results showed that rates of NPK fertilizers significantly affected rapeseed yield. By establishing the fertilizer response equations, the optimum fertilizer rates in oilseed rape were as following, N 118.5kg per hectare, P2O5 36.5kg per hectare, K2O 60.6 kg per hectare for the yield of 2 416.3kg per hectare at Guilin; N 176.3 kg per hectare, P2O5 73.0kg per hectare, K2O100.6kg per hectare for the yield of 3 095.1kg per hectare at Zhijiang, N 256.7 kg per hectare, P2O5 88.1kg per hectare, K2O 137.7kg per hectare for the yield of 4 474.3kg/hm2 at Yulong. It could be concluded that the most important influence factors on optimum rates of fertilizer applied to rapeseed included agro-ecology condition, genotype of cultivar, marginal profit ratio, prices of seeds and fertilizers. It was inaccurate to get the optimum fertilizer rates if they were only based on basic available nutrients of soil.

  • HU Lei, NIU Shi-quan, JING Cai-hong, DA Wen-yan, ZHU Xue-tai, HAN Jian-shan, CHENG Xiao, ZHANG Ai-mei
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    Abstract: Different Streptomyces strains were isolated from saline-alkali soils of Hexi Corridor in Gansu Province. Among them, two strains named Ⅲ22-3-3 and Ⅲ22-3-12 were found to be antagonists inhibiting Sclerotinia sclerotiorum isolated from oilseed rape. Detached leaf inoculation tests showed that the two strains had preventing effects of 63.5% and 49.1% respectively. Moreover, Ⅲ22-3-12 could colonize on the sclerotia and inhibit the germination of the pathogen. Compared to physiological saline (control), Ⅲ22-3-12 decreased 18.2% of the sclerotia germination rate. Morphological and physiological-biochemical characteristics as well as phylogenetic analysis based on 16S rRNA gene sequence supported strain Ⅲ22-3-3 to be Streptomyces hydogenans, and strain Ⅲ22-3-12 to be Streptomyces globisporus.

  • LIU Yue, HUA Xiao-mei, GUO Jing, HU Dong-qing, ZHU Min-min, YANG Tong-yi
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    Bacterial quorum sensing (QS) in the rhizosphere soil of soybean in acid soil was studied with pot and laboratory experiments. N-acyl-homoserine lactone (AHLs) was one of QS signal molecules, it was extracted from the rhizosphere and nonrhizosphere soils of BX10(aluminum -tolerant) and BD2(Al-sensitive). Soybean genotypes cultured in acid soil were detected by Agrobacterium tumefaciens KYC55 (pJZ372; pJZ384; pJZ410). AHLs activities in the rhizosphere soil were much higher than those in nonrhizosphere soil. AHLs activities in the rhizosphere soil of BX10 were much higher than those of BD2 (P<0.01). Moreover, AHLs increased gradually during the process of soybean growth. The results of the laboratory experiments simulating the acid soil environment showed that 3~9umol/L Al significantly influenced the growth, AHLs level, exopolysaccharide production and root hair adherence of Sinorhizobium sp.1128, which further demonstrated the effects of aluminum stress on QS.

  • FUJun-fan, WANG Da-zhou, ZHOU Ru-jun, YANG Feng-yan, SU Wei-na
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    The peanut web blotch disease(Phoma arachidicola Marasas) was one of the serious diseases in Liaoning peanut production and caused serious yield reduction by 30%. This disease damage and epidemic dynamics of peanut web blotch were studied. A systematical investigation in 2011 showed that the percentage of this disease fields was 100%, the percentage of diseased leaf was 32.9% to 66.5%, the disease index was 11.1 to 35.5. It occurred most seriously in Huludao, Fuxin and Jinzhou city, moderately in Shenyang and slightly in Tieling city. The order of disease severity among tested cultivars was Silihong>Baisha1016>Huayu20>Heihuasheng>Baihuasheng. Through comparing and analyzing, Logistic model could well reflect the temporal dynamic of peanut web blotch disease. And the epidemic period could be deduced and the highest disease index increasing rate was 0.99/d. The Silihong was highly susceptible and could be used for disease epidemic studying.

  • WANG Ya,HUANG Si-Liang, HE Peng-Peng, NIU Xiao-Rui,LI Qi-Qin
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    To identify and characterize the pathogen causing sesame southern blight, the pathogen was isolated and its pathogenicity to sesame plants was confirmed based on Koch's postulates. Pathogen identification was carried out based on morphological characteristics and ribosomal DNA internal transcribed spacer (rDNA-ITS) sequences. Partial biological characteristics of the pathogen were tested in vitro. The pathogen causing sesame southern blight was identified as Athelia rolfsii(anamorph: Sclerotium rolfsii). The temperatures suitable for its growth were 22- 34°C with the optimum temperature at 31 °C. A. rolfsii grew well at a pH range of 4.0-9.0 with the optimum pH at 6.5-7.0, indicating that slight acidic to neutral environment was more suitable for vegetative growth of the pathogen. The pathogen could utilize all of the 13 tested carbon sources for its vegetative growth. However, no sclerotia were developed on the media containing each of the seven substances (soluble starch, saccharose, glucose, maltose, D-fructose, L-arabinose and inulin) as a carbon source and nitrate as the nitrogen source. A. rolfsii could produce sclerotia on the media containing glucose as a carbon source and ammonium salt as the nitrogen source. A great number of A. rolfsii sclerotia with larger sizes developed on most of the natural media compared to the synthetic media.

  • CHEN Ran, LI Pei-wu, MA Fei, DING Xiao-xia, ZHANG Wen, ZHANG Qi
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    Equipment was designed for peanut aflatoxins detoxification by ozonation technology. Results showed that factors including ozone accessing methods, ozone concentration, time and peanut moisture, had significant effects on detoxification. The optimal conditions was to input the ozone from the bottom at 6.0mg/L for 30 minutes and exhaust ozone from top using peanut with 5%. Moisture level. Under the given conditions, the detoxification effects of total aflatoxins and aflatoxin B1 were 65.88% and 65.90% respectively. Results indicated a simple, reproducibility and environment friendly technique, suitable for peanuts aflatoxins detoxification.

  • LIU Chang-sheng, YANG Mei, HUANG Feng-hong, ZHOU Qi, ZHENG Chang
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    Adsorption behavior of sodium montmorillonite (Na-MMT) was evaluated on degumming of rapeseed oils. Na-MMT was prepared with calcium montmorillonite by sodium-modification, and was analyzed with X-ray diffraction and thermal gravimetric. The static adsorption equilibrium, adsorption kinetics and thermodynamics of phospholipids (PL) on Na-MMT were also investigated. Results showed that the theoretical maximum adsorption of PL on Na-MMT was 833.3mg/g. The PL adsorption process was favorable, endothemic and spontaneous. Freundlich model fitted well to the adsorption isotherm, and pseudo-second order kinetics model described well on the PL adsorption kinetics

  • CHEN Meng,YANG Mei, LIU Chang-sheng, HUANG Feng-hong
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    Rapeseed polyphenols is an important active substance of rapeseed with diverse biological activities including antitumor, hypoglycemic, antibacterial, antioxidant, scavenging free radicals. Most of the polyphenols remained in rapeseed meal after pressing, which limited the utilization of rapeseed meal. In this paper, the methods of extraction, purification and detection of rapeseed polyphenols were reviewed, and polyphenols’ changes during seed oil processing was also discussed.

  • SUN Mei-yu,HUA Wei,LIU Jing,WANG Xin-fa,LIU Gui-hua,WANG Han-zhong
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     Effective silique number on main inflorescence is an important factor on rapeseed (Brassica napus L.) yield. Doubled haploids (DH) derived from ZY036 and 51070 were used to develop genetic markers on effective silique number. DH population was planted for 3 years at 4 locations as Wuhan-2010, Yangluo-2011, Qinghai-2011, Yangluo-2012 and Xiangyang-2012 respectively. In the 5 field trials, QTLs for the effective silique number on main inflorescence were identified using WinQTL Cartographer 2.5 software. 10 QTLs were found. Their phenotypic variation,LOD value and additive effect ranged from 9.33% to 31.60%,2.51 to 5.37 and 2.71 to 8.09 respectively. Four QTLs on chromosomes A1, A5, C1 and C9 were co-localized by 2 trials. In the QTL regions, 16 linkage markers were obtained, and 8 of 16 markers had less than 1cM distance from the peak of the QTLs.