Key Laboratory of Oil Crop Biology of the Ministry of Agriculture

Wuhan, China

Key Laboratory of Oil Crop Biology of the Ministry of Agriculture

Wuhan, China
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Guan D.,Chinese Academy of Agricultural Sciences | Guan D.,Key Laboratory of Oil Crop Biology of the Ministry of Agriculture | Guan D.,Quality Inspection and Test Center for Oilseeds Products | Li P.,Chinese Academy of Agricultural Sciences | And 6 more authors.
Analytica Chimica Acta | Year: 2011

A green enzyme-linked immunosorbent assay (ELISA) to measure aflatoxin M 1 (AFM 1) in milk was developed and validated with a surrogate calibrator curve. Polyclonal anti-idiotype (anti-Id) antibody, used as an AFM 1 surrogate, was generated by immunizing rabbits with F(ab′) 2 fragments from the anti-AFM 1 monoclonal antibody (mAb). The rabbits exhibited high specificity to the anti-AFM 1 mAb, and no cross-reactivity to either of the other anti-aflatoxin mAbs or the isotype matched mAb was observed. After optimizing the physicochemical factors (pH and ionic strength) that influence assay performance, a quantitative conversion formula was developed between AFM 1 and the anti-Id antibody (y=31.91x-8.47, r=0.9997). The assay was applied to analyze AFM 1 in spiked milk samples. The IC 50 value of the surrogate calibrator curve was 2.4μgmL -1, and the inter-assay and intra-assay variations were less than 10.8%; recovery ranged from 85.2 to 110.9%. A reference high-performance liquid chromatography method was used to validate the developed method, and a good correlation was obtained (y=0.81x+9.82, r=0.9922). © 2011 Elsevier B.V.


Sha A.,Chinese Academy of Agricultural Sciences | Sha A.,Key Laboratory of Oil Crop Biology of the Ministry of Agriculture | Chen Y.,Hainan University | Ba H.,Chinese Academy of Agricultural Sciences | And 10 more authors.
Journal of Plant Biology | Year: 2012

MicroRNAs (miRNAs) are endogenous small RNAs regulating plant development and stress responses. In addition, phosphorus (P) is an important macronutrient for plant growth and development. More than two hundred miRNAs have been identified in Glycine Max and a few of miRNAs have been shown to respond to P deficiency, however, whether there are other miRNAs involved in P deficiency response is largely unknown. In this study, we used high-throughput small RNA sequencing and whole-genome-wide mining to identify the potential miRNAs in response to P deficiency. After sequencing, we deduced 183 known, 99 conserved and 126 novel miRNAs in Glycine Max. Among them, in response to P deficiency, the expressions of 27 known, 16 conserved and 12 novel miRNAs showed significant changes in roots, whereas the expressions of 34 known, 14 conserved and 7 novel miRNAs were significantly different in shoots. Furthermore, we validated the predicated novel miRNAs and found that three miRNAs in roots and five miRNAs in shoots responded to P deficiency. Some miRNAs were P-induced whereas some were P-suppressed. Together these results indicated that the miRNAs identified might play important roles in regulating P signaling pathway. © 2012 Korean Society of Plant Biologists and Springer-Verlag Berlin Heidelberg.


Ai-Hua S.,Chinese Academy of Agricultural Sciences | Ai-Hua S.,Key Laboratory of Oil Crop Biology of the Ministry of Agriculture | Yin-Hua C.,Hainan University | Zhi-Hui S.,Chinese Academy of Agricultural Sciences | And 9 more authors.
Journal of Genetics | Year: 2014

Soybean (Glycine max) is a short-day crop and the photoperiod is a crucial factor regulating its flowering time. To investigate the molecular mechanism controlling the flowering time by photoperiod in soybean, cDNA-amplified fragment length polymorphism (cDNA-AFLP) was used to identify photoperiod-regulated genes in leaves of soybean growing under short-day length, neutral photoperiod and long-day length. A total of 36 transcript-derived fragments (TDFs) were identified to be regulated by photoperiod. Among them, 26 TDFs were homologues of genes with known function. These genes are involved in secondary metabolism, cellular metabolism, cell wall components metabolism, ion transport and hormone signalling. Silencing of the homologue genes in Nicotiana benthemiana for 14 TDFs was conducted by virus-induced gene silencing. The flowering time was delayed by silencing of the genes encoding rhodanese and 40S ribosomal protein S4 (RPS4). The results indicated that rhodanese and RPS4 probably play important roles in regulating flowering time. © 2014 Indian Academy of Sciences.


Guan D.,Chinese Academy of Agricultural Sciences | Guan D.,Key Laboratory of Oil Crop Biology of the Ministry of Agriculture | Guan D.,Quality Inspection and Test Center for Oilseeds Products | Li P.,Chinese Academy of Agricultural Sciences | And 10 more authors.
Food Chemistry | Year: 2011

A sensitive and specific monoclonal antibody (Mab) against aflatoxin M1 (AFM1), named as 2C9, was selected by semi-solid HAT medium. It exhibited high affinity for AFM1 of 1.74×109L/mol and no cross-reactivity to aflatoxin B1, B2, G1 and G2. Based on the antibody, an ultra-sensitive competitive enzyme-linked immunosorbent assay (ELISA) was developed for AFM1 in milk and infant milk products. Assays were performed in the AFM1-BSA coated (0.0625μg/mL) ELISA format in which the antibody was diluted 1:10,000. Several physicochemical factors (pH, ionic strength and blocking solution) that influence assay performance were optimised. Finally, the limits of detection were 3ng/L for milk and 6ng/L for milk-based cereal weaning food, inter-assay and intra-assay variations were less than 10%, and the recovery ranged from 91% to 110%. Thirty samples were analysed, and concordant results were obtained when the data were compared with a reference high-performance liquid chromatography method. © 2010 Elsevier Ltd.


Wang Y.,Chinese Academy of Agricultural Sciences | Wang Y.,Key Laboratory of Biotoxin | Wang Y.,Quality Inspection and Test Center for Oilseeds Products | Zhang Q.,Chinese Academy of Agricultural Sciences | And 11 more authors.
Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences | Year: 2011

This paper describes the establishment of an immunoaffinity chromatography (IAC) for selective extraction of fenvalerate from vegetable samples. The IAC column was constructed by covalently coupling monoclonal antibody (mAb) against fenvalerate to CNBr-activated Sepharose 4B and packed into a cartridge. The extraction conditions were carefully optimized, including loading, washing and eluting solutions. Under the optimal conditions, the IAC column was able to capture fenvalerate with the maximum capacity of 4000ng. An average recovery of 94.5% and a RSD of 8.8% were obtained with six IAC columns prepared on six different days. Three vegetable samples spiked with fenvalerate at four different concentrations were extracted with IAC column and determined by gas chromatography with electron capture detection (GC-ECD). Chromatograms of final extracts were clean and fenvalerate could be easily detected without the interferences. The extraction recoveries and RSD were 74.7-96.5% and 2.5-5.2%, respectively, and the calculated limit of detection of the whole method was 0.008-0.012ngg -1. © 2011 Elsevier B.V.


Zhang D.,Chinese Academy of Agricultural Sciences | Zhang D.,Key Laboratory of Oil Crop Biology of the Ministry of Agriculture | Zhang D.,Quality Inspection and Test Center for Oilseeds Products | Li P.,Chinese Academy of Agricultural Sciences | And 6 more authors.
Biosensors and Bioelectronics | Year: 2011

An ultrasensitive immunochromatographic (IC) assay for simultaneous detection of total aflatoxins (AFB1, AFB2, AFG1, and AFG2) was developed to meet the requirement for rapidly monitoring aflatoxins in agro-products. The assay was based on a competitive format and its sensitivity was improved by using a novel criterion to screen the optimal amount of monoclonal antibody (MAb) labeled to nanogold particles. The visual detection limits (VDLs) for aflatoxins B1, B2, G1, and G2 in peanut matrix were 0.03, 0.06, 0.12, and 0.25ngmL-1, respectively, which were lower than those of published literatures. The results of IC assay were in good agreement with those of high performance liquid chromatography (HPLC) in the analysis of aflatoxins in peanuts, demonstrating the practical applicability of the developed assay in real samples. This qualitative test based on the visual evaluation of results did not require any equipment. Overall, to our knowledge, this is the first report of qualitative detection for total aflatoxins by immunochromatographic assay. © 2010 Elsevier B.V.


Jiang J.,Chinese Academy of Agricultural Sciences | Zhang D.,Chinese Academy of Agricultural Sciences | Zhang W.,Chinese Academy of Agricultural Sciences | Zhang W.,Quality Inspection and Test Center for Oilseeds | And 8 more authors.
Analytical Letters | Year: 2010

Monoclonal antibodies (MAbs) against pyrethroid insecticide fenvalerate were achieved, identified, and applied in environmental water. Mice were immunized with a novel synthesized hapten conjugated with bovine serum albumin (BSA). Three positive clones of MAbs were obtained after cell fusion and hybridoma selection, among them MAb-2 (5B10) showed the highest reactivity toward fenvalerate. The IC50 of MAb-2 was 94.5 ngmL-1; moreover, it showed lower cross-reactivity with other pyrethroids such as bifenthrin, tetramethrin, deltamethrin and beta-cypermethrin. Optimization of enzyme-linked immunosorbent assay (ELISA) was studied. The limit of detection (LOD) of the assay was 8.8 ngmL-1 and the detection range was 0.017-27.33 μgmL-1. For preliminary application, addition recovery experiments in water samples were performed. The mean recoveries of three kinds of samples varied from 90.6% to 108.7% and the coefficients of variation ranged from 0.5% to 5.3%. The results showed that MAb-2 could be used for the detection of fenvalerate contamination in environmental water. © Taylor & Francis Group, LLC.


Guo B.,Chinese Academy of Agricultural Sciences | Jiang M.,Chinese Academy of Agricultural Sciences | Jiang M.,Key Laboratory of Oil Crop Biology of the Ministry of Agriculture | Wan X.,Chinese Academy of Agricultural Sciences | And 4 more authors.
Journal of Microbiology and Biotechnology | Year: 2013

The marine microalga Isochrysis sphaerica is rich in the very-long-chain polyunsaturated fatty acids, including eicosapentaenoic acid (EPA, C20:5ω-3) and docosahexaenoic acid (DHA, C22:6ω-3) that are important to human health. Here, we report a functional characterization of a &δ4-fatty acid desaturase gene (FAD4) from I. sphaerica. IsFAD4 contains a 1,284 bp open reading frame encoding a 427 amino acid polypeptide. The deduced amino sequence comprises three conserved histidine motifs and a cytochrome b5 domain at its N-terminus. Phylogenetic analysis indicated that IsFad4 formed a unique Isochrysis clade distinct from the counterparts of other eukaryotes. Heterologous expression of IsFAD4 in Pichia pastoris showed that IsFad4 was able to desaturate docosapentaenoic acid (DPA) to form DHA, and the rate of converting DPA to DHA was 79.8%. These results throw light on the potential industrial production of specific polyunsaturated fatty acids through IsFAD4 transgenic yeast or oil crops. © 2013 by The Korean Society for Microbiology and Biotechnology.


Wang P.,Chinese Academy of Agricultural Sciences | Wan X.,Chinese Academy of Agricultural Sciences | Wan X.,Key Laboratory of Oil Crop Biology of the Ministry of Agriculture | Zhang Y.,Chinese Academy of Agricultural Sciences | And 2 more authors.
Biotechnology Letters | Year: 2011

A novel expression system was established in the oleaginous yeast, Lipomyces kononenkoae. The expression vector pLK-rhPHG of L. kononenkoae was constructed and using the hygromycin phosphotransferase gene and green fluorescent protein gene as reporter genes. A delta 6-fatty acid desaturase gene (D6DM) from Cunninghamella echinulata MIAN6 was then expressed in this strain. The recombinant strain accumulated about 1.2% γ-linolenic acid in the total fatty acids. © 2011 Springer Science+Business Media B.V.


Wu D.,South China Agricultural University | Zhao M.,South China Agricultural University | Shen S.,South China Agricultural University | Fu Y.,South China Agricultural University | And 4 more authors.
Acta Physiologiae Plantarum | Year: 2013

Al-activated organic acid transporter genes (ALMT and MATE) and plasma membrane H+-ATPase gene (PHA) are known to contribute to the regulation of organic acid secretion in several crops. However, it remains unclear how these genes interact to modulate organic acid exudation in the same plant species. In this study, Al-induced expression of genes (GmALMT1, GmMATE1 and GmPHA1), secretion of organic acid and root elongation were characterized in soybean roots. Results indicated that treatment with 50 μM Al activated the expression of GmALMT1, GmMATE1 and GmPHA1, and the exudation of citrate and malate significantly in apical 5 mm region of soybean seedlings, but inhibited root elongation by 57. 8 %. The highest malate exudation rate and the maximal expression of GmALMT1 and GmPHA1 were observed after 2 h of 50 μM Al treatment, while the corresponding values for citrate exudation rate and GmMATE1 expression occurred at 8 h. The exudation of malate and citrate contributed to but could not recover Al-triggered root elongation. A root-split experiment indicated that Al-activated gene expression, organic acid secretion and root growth inhibition required the direct contact of Al3+. The removal of shoots in soybean seedlings decreased Al-activated gene expression by 26. 1-40. 5 %, and secretion of organic acid by 14. 7-40. 2 %. Furthermore, shoot excision aggravated Al-inhibited root elongation, indicating the existence of other Al tolerance mechanism except the exudation of organic acid. These results suggested that Al-activated expression of GmPHA1-, GmMATE1- and GmALMT1-mediated exudation of malate and citrate, and shoots played an important role in Al toxicity resistance in soybean roots. © 2012 Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków.

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