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  • 2016 Volume 38 Issue 2
    Published: 28 April 2016
      

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  • MA Li, SUN Wan-cang, LIU Zi-gang, FANG Yan, WU Jun-yan, LI Xue-cai, LI Xin, ZHANG Shu-juan, YUAN Jin-hai, CHEN QI, WANG Kai-yin, FANG Yuan
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    To understand different cold-tolerance of Brassica rapa and B. napus, 3 genes were investigated under chilling stresses at 4℃, -4℃ and -8℃. The genes included BnCOR25 (from B. napus), BnICE1 (from B. napus), and Cu/Zn-SOD (from B. rapa). 6 different cold-resistant winter rapeseeds were used as materials and analyzed after 5d chilling treatments and normal control (at 24℃, CK). qRT-PCR results showed that BnCOR25, BnICE1 and Cu/Zn-SOD were not activated and expressed at the same level in both B. rapa and B. napus at 24℃. Under chilling, their relative expressions in B. rapa increased when the temperature decreased, and reached to the peak at -4℃. Among them, BnCOR25 reached the peak at 4℃ and Cu/Zn-SOD at -4℃ in B. napus. BnICE1 expression was always lower than CK. At -8℃, genes of both types were decreased and low. Compared to B. napus, these 3 genes in B. rapa were significantly higher. Compared to cold-sensitive rapeseed, those with higher cold tolerance had higher expression. It indicated that cold-tolerance was related to these 3 genes and their different responses. 

  • XU Ben-bo, XIE Ling-li, LI Jia-na, CHAI You-rong*, ZENG Liu
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     To investigate the relationship between BnCHI-1 gene and rapeseed color, a BnCHI-1 gene was cloned from B. napus stock line 5B by RACE amplification. The genomic sequences of BnCHI-1 was 1 809 bp and contained 3 introns. The 965 bp BnCHI-1 mRNA had a 756 bp ORF encoding 251 amino acids. NCBI conserved domain search indicated that BnCHI-1 gene contained conserved gnl|CDD|8301 domain (chalcone-flavanone isomerase), (2S)-naringenin binding cleft and residues of the active site hydrogen bond network. Southern blot result indicated that it's likely that 6 CHI genes existed in B. napus. RT-PCR indicated that the tRNA BnCHI genes were ubiquitously expressed in 11 detected organs of B. napus line 5B. Relatively higher expression levels were detected in stems, leaves, buds and flowers, followed by seeds and roots, and the cotyledons had the lowest expression level. No significant difference of the total transcription of BnCHI family was detected between L1 and L2 (a pair of near-isogenic lines, black line and yellow line respectively) in buds, flowers, 20 DAF seeds and 30 DAF seeds. But in 10 DAF seeds of L2, the expression was significantly lower than that of L1.

  • ZHANG Yan-wei, LI Wei, ZHANG Li-feng, WANG Cai-jie, DAI Hai-ying, XU Ran*
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    For an overall understanding of the whole genome of important soybean breeding parent and elite cultivar Qihuang 34, high-throughput sequencing technology was used by re-sequencing with 13× depth to detect its variations. Results revealed 1 519 494 single nucleotide polymorphisms (SNPs), 357 549 small Indels and 4 506 structure variations. They confered 17 748 DNA variations. These variations included key photoperiod related genes, such as CRYPTOCHROME 2, GIGANTEA, Timing of CAB expression 1, E1, PHYTOCHROME A, Flowering Locus T, CONSTANS, Terminal Flower like, et al. Results indicated that Qihuang 34 was E1 genotype. The above SNPs, Indels and structure variations would become important markers for marker-assisted selection for soybean breeding.

  • HUANG Yi-an, DENG Xiao-juan, WAN Hai-bo, WANG Peng, FANG Xiao-long, ZHANG Jie, YANG Cun-yi
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    To provide better materials for breeding and functional gene study, mutagenesis was carried out on soybean cultivar Huaxia 3 by ethyl methane sulphonate (EMS) and seed irradiation with 60Co gamma-ray. Phenotypic traits including leaf, stem, flower, seed, pod and flowering-time variations were investigated in M4 generation. 86 mutants were identified from the population including mutants on leaf (13), stem (30), flower color (4), seed (2), pod (15), and flowering-time (22). 82 pairs of SSR markers were used to screen the genomic difference between wild type and 270 mutants. Among them, 4 mutants (M3-91, M3-500-24, EMS3-23 and EMS3-31) had more than 10 polymorphic markers. 6 mutants were different to wild type on locus Satt513. 

  • LI Chao,SHA Ai-hua*
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    MicroRNA play regulation roles in phosphorus signal pathway. miR319 was detected up-regulated in soybean roots under phosphorus deficiency. For more understanding of this gene, GmMIR319 was cloned from soybean and transformed into tobacco. Low phosphorus response of miR319 was investigated in overexpressing transgenic tobacco. Results showed that transgenic tobacco root was longer than that of wild type, and fresh weights of roots and shoots were higher in transgenic plants under phosphorus deficiency. It indicated that overexpression of GmMIR319 in tobacco improved tolerance to phosphorus deficiency. TC18729, one of the putative target of miR319, was down-regulated in transgenic plants when miR319 was up-regulated. It suggested that miR319 responses in phosphorus signal pathway in tobacco by regulating TC18729 expression. 

  • LIU Hua, QIN Li, ZHANG Xin-you*, HAN Suo-yi, DU Pei, ZHANG Zhong-xin, SUN Zi-qi, QI Fei-yan, DONG Wen-zhao, HUANG Bing-yan
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    To better understand the genetic mechanism of fatty acid traits and to facilitate peanut quality enhancement, genetics of a F10 population derived from interspecific cross Baisha 1016×A. monticola was analyzed using major gene plus polygene mixed inheritance model. 216 recombinant inbred lines (RILs) were investigated to build a genetic model. Results showed that contents of both fat and arachic acid were controlled by polygenes, and their heritability were 98.55% and 84.81% respectively. Each with 5 components including oleic acid, linoleic acid, palmitic acid, behenic acid and lignoceric acid, were controlled by 2 major genes plus polygenes. Their major gene heritability were 45.08%, 46.13%, 44.85%, 66.55% and 65.98% respectively. Correlation analysis showed that both oleic acid and arachic acid contents were highly significantly (positive) correlated to peanut fat content. Linoleic acid, palmitic acid, behenic acid and lignoceric acid contents were highly significantly (negative) correlated to peanut fat content. Therefore, high-oil peanut breeding should focus on polygenes accumulation. And high oleic acid and other major fatty acid breeding should focus on both major genes and polygenes.

  • HOU Wen-huan, LIN Jing, YAN Long, YANG Yong-qing,WANG Tao, YANG Chun-yan, ZHI Hai-jian, ZHANG Meng-chen*
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    To understand soybean gene inheritance and allelism resistant to recombined-SMV isolate (HB-RS), crosses were made between resistant (R) and susceptive (S) cultivars. Results showed that F1 plants from R×S crosses (Jidou 12×Franklin, Jidou 17×10Y105, Jidou 12×PI632401, PI96983×ZYD2738 and Wuxing 4×FH13) were resistant, and their F2 segregated in 3R:1S or 15R:1S. In R×R crosses, both F1 and F2 plants of crosses Jidou 12×Newton and Jidou 12×PI96983 were resistant. F1 plants of Jidou 12×V94-5152 were resistant, and F2 segregated in 15R:1S. In other crosses, F1 plants of crosses Jidou 17×Newton, PI96983 and V94-5152 were resistant and F2 segregated in 15R:1S. The results suggested that each of the cultivars (Jidou 12, Jidou 17 and PI96983) carried 1 dominant resistant gene for HB-RS, and Wuxing 4 carried 2 dominant genes. The resistance gene from Jidou 12 was allelic or closely linked to those from Newton and PI96983, but not allelic with that from V94-5152. The resistant gene from Jidou 17 was not allelic with those from Newton, PI96983 and V94-5152.

  • WANG Li-min ZHANG Jian-ping, DAGN Zhao, DANG Zhan-hai*
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    To further understand flax temperature sensitive male sterile lines 1S and 113S, 2 crosses with Longya 10 (oil flax) and Heiya 15 (fiber flax) were used by constructing P1, P2, F1 and F2 populations for each cross. Their genetic foundations were investigated on 4 yield-related traits using mixed-major-genes-plus-polygenes inheritance model. Results indicated that yield per plant, capsules per plant and 1 000-seed weight were controlled by 2 major genes with additive-dominance-epistatic effects plus polygenes with additive-dominance effects. Seeds per capsule were controlled by 2 major genes with additive-dominance effects plus polygenes with additive-dominance effects in the 2 crosses. In F2 populations, the major gene heritability of yield per plant, capsules per plant and seeds per capsule in the 2 crosses were 43.50% to 73.28%, those of 1 000-seed weight were 10.55% to 34.40%. It indicated that additive, dominance and epistatic effects of major genes played important roles in yield inheritance in the flax temperature sensitive male sterile lines. To increase hybrid flax production, additive and dominant effects should be utilized in two-line heterosis breeding in flax.

  • HE Qing-yuan, XIANG Shi-Hua, LI Zheng-peng, WANG Song-hua, ZHU Chang-wei, LIU Bin, WU Ping*,
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    H2S could alleviate the salt stress of soybean. For explore the mechanism of H2S in relieving NaCl stress, 0.08 mol·L-1 NaCl was used in hydroponics experiments for soybean seedlings (Nannong1138-2) at V2 period. Different concentration of NaHS (in 0.00, 0.20, 0.40, 0.60 and 0.80 mmol·L-1) were sprayed every 24h. Effect of H2S on methylation level, degree and status of soybean in V3 and V4 periods under salt stress were analyzed using methylation sensitive amplified polymorphism (MSAP) method. Results showed that methylation level of soybean was decreased by NaCl stress except for exterior hemi-methylation in V4 period. However, after NaHS spray, soybean methylation level under NaCl for 2 periods were initially increased and then decreased in a certain concentration range except for inner fully methylation in V3 period. It suggested that H2S alleviated soybean salt damage by restoring both level and type of methylation.

  • TIAN Shi-gang, CHEN Jun-kun, SHEN Qi, WANG Xian-ping, GUO ping
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    For low cost and high yield cultivation, practised cultivation methods of perilla were studied under 5 different cultivations. Results showed that manual seedling transplantation mode has the highest yield (1 675.5 kg/hm2), but with the highest unit cost (in RMB yuan) of 15 924 yuan/hm2, and the lowest economic benefit (10 884.0 yuan/hm2). For the 2 methods of mechanical involvement, mechanical ploughing and sowing, and mechanical ploughing and manual sowing, the yields were 1 372.4 kg/hm2 and 1 317.3 kg/hm2, with unit costs of 5 202 and 5 952 yuan/hm2, and economic benefits of 16 756.4 and 15 124.8 yuan/hm2 respectively, which were significantly better than the other 3 methods. Among these 2 methods, mechanical ploughing and sowing was more suitable for larger scale of production, and mechanical ploughing and manual sowing was more suitable for mountain, slope and smaller areas. Both were ideal methods for perilla. In addition, all 5 cultivation methods had great effects on effective branch, effective panicles and seeds per panicle on the main inflorescence. These methods had less effect on plant height, thousand seeds weight and seed oil. 

  • WU Zheng-feng,ZHENG Yong-mei, CHEN Dian-xu,, WANG Cai-bin, SUN Xue-wu, LI Xiang-dong, WANG Xing-xiang, SHI Cheng-ren, FENG Hao, YU Tian-yi
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     To determine economic nitrogen (N) supply and peanut N use efficiency during various growth stages, non-nodulated variety (NN-1) and nodulated variety (Huayu 22) were used in pot experiments with 15N tracing to investigate N supply sources (from soil, root nodule and N fertilizer) and N use efficiency at different stages in 2010 and 2011. Results showed significant differences in 3 N supply sources at different stages. N supply rate from root nodule was shown as unimodal curve with the peak at podding phase. Higher supply rate of N from soil was at pegging and pod filling phases, and N supply from fertilizer was higher at pegging and podding phases. With regard to the contributions of N supply sources, N from root nodule showed an unimodal change with the peak at podding phase, and N from soil appeared an irregular trait “V” with the minimum value at podding phase, and N from N fertilizer gradually declined. The total N supply sources over the whole growth stage mainly came from root nodule, soil and N fertilizer. The N supply proportion from root nodule, soil and N fertilizer was 5:3:2. The N use efficiency was above 40.0% for the overall growth period. Significant differences were shown among the individual growth stages and ranked in the order of podding > pegging > seedling > pod filling. In conclusion, over the entire growth period, root nodule was the greatest N source accounted for about 50.0%, and soil supplied higher N than N fertilizer for peanut. N use efficiency over the entire growth period was determined by N uptake at individual growth stage, which was mainly come from podding phase, closely followed by pegging phase, with the lowest from pod filling phase.

  • GAO Jing, ZHANG Yuan-yuan, WANG Kai, ZHANG Jian, ZHANG Gui, ZHAO Jun?
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    To identify the pathogen of sunflower wilt disease in China, samples were collected from major Chinese planting area in 2014-2015. 50 diseased plants from different sites were collected and identified via Koch’s postulate. 31 purified isolates were identified as 7 species of Fusarium genus. They were F. oxysporum, F. verticilloides, F. lateritium, F. acuminatum, F. redolens, F. equiseti and F. proliferum. Representative strain was randomly chosen from these 7 different species respectively for identifying their biological characteristics. Results showed that growth speed of F. verticilloides BC012 was the fastest, and F. lateritium BC03 was the slowest. BC012 also had the highest yield of conidia, while F. equiseti NMG1 had the least. The most suitable temperature for all 7 strains were between 25℃ to 30℃, except that F. acuminatum TBB212 could grow under 5℃. The medium pH ranged from 4 to 11. The crude toxin yields were 0.651 mg/mL (by F. oxysporum BC102) to 0.075 6 mg/mL (by F. lateritium BC03). Pathogenicity test results showed that disease index of BC012, WLJ1 (F. redolens) and TBB212 were 65.0, 55.0 and 55.0 respectively after 21d post-inoculation (21 dpi) with early leave chlorisis. For the other 4 species, F. equiseti NMG1, F. proliferum T5, F. oxysporum BC102 and F. lateritium BC03, the disease index ranged between 30.0 and 40.0 with slower infection and fewer diseased leaves. For control strain of Verticillium dahliae, which caused sunflower yellow wilt, the index was only 25.0.

  • HUI Hong-yan, WANG Yuan-yuan, ZHU Xiao-feng, CHEN Li-jie, LIU Xiao-yu, ZHANG Meng-yue, DUAN Yu-xi
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     It is of great importance to make use of endophytes in antagonism controlling on Meloidogyne.In order to obtain a wide variety of endophytic bacteria strains, soil samples were collected from different ecological environment to trap peanut nodules. Endophytes were separated from peanut nodule by using half-of-a-nodule method, and endophytic bacteria strains with higher toxicity to 2-year-old larva of root knot nematodes were selected.Among 637 nodule samples, 19 efficient nodule had higher lethal rate over 80%. The highest lethal rate was over 94% after 24h. 123 endophytic bacteria strains were separated and purified from above 19 nodule samples, and their fermentation nematicidal activities were compared.Results showed that, 9 bacterial strains had higher activity, especially Sneb1706 could cause 91.4% mortality of J2 after 24h, and Sneb1872 had an incubation inhibition ratio of 59.4%.Meanwhile, Sneb1773 showed obvious growth promoting effect on plants.16S RNA sequence analysis, and their physiological and biochemical identification indicated that these 9 strains of nodule endophytes belong to Pseudomonas, Rhizobium and Stenotrophomonas. 

  • LIU Wei-guo, ZOU Jun-lin, YANG Chen-yu, LUO Ling, JIANG Tao, YANG Wen-yu*
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    To optimize soybean stem protein extraction and separation method for better application of 2-D electrophoresis on soybean proteomics, extraction method, pH of IPG strips, sample loading quantity, SDS-PAGE gel concentration and staining method were compared. Results showed that the new protein extraction method combining TCA/acetone precipitation and phenol extraction could resolve more protein with better separation by 2-D electrophoresis. The optimized processes included pH3-10 non-linear IPG strip and loading 800 μg soybean stem protein for the first-dimensional IEF, 12% polyacrylamide gel for the SDS-PAGE electrophoresis, and Coomassie brilliant blue staining.

  • ZHANG Xin-ran,LI Pei-wu*,DING Xiao-xia,WU Wen-hua
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    To provide a reference for peanut quality safety risk monitoring and risk assessment, 604 peanut samples were collected from 8 provinces and evaluated on their mycotoxin infections by risk ranking method. According to obtained data and limiting standard, the hazard degrees of fungus toxin risk factor in these peanut samples were identified. The effective evaluating indicators were chosen, and a normalized system was established for searching, identifying and risk ranking on peanut mycotoxin risk factors. 

  • ZHANG Peng, JIN Fen*, LI Min-jie, JIANG Ze-jun, WU Ri-na, SHAO Hua, JIN Mao-jun, SHE Yong-xin, ZHENG Lu-fei, WANG Shan-shan, WANG Jing
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     To explore the pollution characteristics of HBCD (hexabromocyclododecane) isomers and TBBPA (tetrabromobisphenol) contamination levels and composition characteristics, 7 vegetable oils in Beijing supermarkets were sampled and analyzed by liquid chromatography-electrospray ionization source-tandem mass spectrometry (LC-ESI-MS/MS). The LOQ of this method is 0.004~0.04 µg/kg. The total level of HBCD and TBBPA detected were LOQ to 0.349 µg/kg and LOQ to 0.222 µg/kg respectively. The detected rates were 56.25% and 50%. Both level and rate were relatively lower than those published. Dietary risk assessment was performed using margin of exposure (MOE). Results showed that MOE of HBCD and TBBPA were 78 000.28 and 126.29 respectively, which were much higher than 8 and 10, (recommended by European Food Safety Authority, eg. EFSA). It indicated that pollution level and dietary risk of total HBCD and TBBPA in sampled vegetable oils in Beijing supermarket were acceptable. 

  • WANG Wei-jun, LIU Chang-sheng, HUANG Feng-hong, LI Wen-lin*, YANG Bo
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    For better deacidification of tea seed oil, alkaline microcrystalline cellulose (AMC) was modified by solid-phase synthesis with microcrystalline cellulose (MCC) and sodium hydroxide (NaOH). AMC and MCC were analyzed by Fourier infrared spectrum, X-ray diffraction, thermal gravimetric analyzer and scanning electron microscope. The effect of AMC on removal of free fatty acids (FFA) from cold-pressed tea seed oil was also studied. Results showed that compared with MCC, crystallinity, hydrogen bonding force and thermal stability of AMC were decreased, while its surface loose degree and porosity increased, and adsorption capacity increased. The optimal deacidification conditions were determined through single-factor and orthogonal experiments. The optimum deacidification time was 2 hours at 45℃, and AMC amount was 2.5%. Under the above conditions, acid value of tea seed oil could be reduced by 3.15 mg KOH/g.

  • ZENG Liu, HU Cheng-wei, ZHANG Xue-kun, CHENG Yong, LU Guang-yuan, ZOU Xi-ling*
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    To find out the optimal colchicine concentration in the main planting area along the Yangtze River for rapeseed (Brassica napus L.) diploid regeneration, 4 colchicine levels were investigated using 2 varieties. Results showed that the highest embryogenesis frequency was obtained at 24 h treatment of 200 mg/L colchicine. Using this treatment, 40 germplasm accessions were studied by investigating genotypic differences in embryogenesis. Results revealed that embryogenesis frequencies were significant different, which ranged from 0.12 to 10.39 (embryos per bud). The efficiencies of chromosome doubling were also different (which ranged from 26.7% to 90%) and showed genotype dependency. When classified by seed color, significant differences were found between yellow and black materials on embryogenesis frequency, but not on chromosome doubling efficiency. This result suggested that embryogenesis efficiency was probably affected by testa color of rapeseed.

  • ZHU Wei, LIU Qing2 YANG Shou-xiang*
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    To define differences of selenium accumulation ability between different peanut cultivars, effects of fertilizing selenium on peanut grain yield and selenium accumulation ability were investigated by method of fertilizing soil with selenium in blooming period and the early stage of podding with pot experiments. 18 peanut cultivars promoted in the main peanut production areas along Yellow River were used as materials. Results showed that compared with control, selenium fertilizer led to different yield, different selenium accumulation and its final content in grain. Compared with no selenium fertilizer, 3 mg Se (IV)/kg soil led to average 0.06% yield loss of 18 cultivars. But their selenium content, selenium cumulant and selenium use efficiency of grain increased by 36.84%, 32.66% and 19.62% respectively. And 6 mg Se (IV)/kg soil led to average yield increase of 18 cultivars by 8.65%. Among them, cultivar Fenghua 5 had the maximum yield increase. Average selenium content, selenium cumulant and selenium use efficiency increased by 35.80%, 36.31% and 35.42% respectively. It indicated that 6 mg Se (IV)/kg fertilizer was optimal. Cluster analysis results (based on yield, selenium content and grain selenium cumulant) showed that 7 cultivars had higher selenium accumulation ability, 7 had medium ability and 4 had low ability.

  • YANG Jun-fang, WANG Ya, ZHANG Huan-huan, WANG Yi-xue,HAO Yao-shan,GUO Zhi-qiang*,WANG Hong-wei*
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    To improve the castor (Ricinus communis L.) gene transformation, imbibed seeds (germinating embryos) were transformed with herbicide resistant gene CP4-EPSPS by Agrobacterium-mediated transformation without tissue culture. Result showed that pricking 3 stitches at meristematic region (apical point) of germinating seeds followed by ultra-sonication (800 W), and co-cultured with Agrobacterium for 16 h were the optimum transformation conditions. 5 putative transgenic plants were detected from 800 randomly-selected treated seedlings, with average positive rate of 0.63%. After glyphosate screening, 3 positive T0 seedlings were obtained out of 26 survivals by PCR and Southern blot. This method was much simpler and more effective than the procedure of tedious tissue culture.