1.Emerging role of ER quality control in plant cell signal perception.
Protein & Cell 2012;3(1):10-16
The endoplasmic reticulum quality control (ER-QC) is a conserved mechanism in surveillance of secreted signaling factors during cell-to-cell communication in eukaryotes. Recent data show that the ER-QC plays important roles in diverse cell-to-cell signaling processes during immune response, vegetative and reproductive development in plants. Pollen tube guidance is a precisely guided cell-cell communication process between the male and female gametophytes during plant reproduction. Recently, the female signal has been identified as small secreted peptides, but how the pollen tube responds to this signal is still unclear. In this review, we intend to summarize the role of ER-QC in plants and discuss the recent advances regarding our understanding of the mechanism of pollen tube response to the female signals.
Animals
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Endoplasmic Reticulum
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metabolism
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Humans
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Plant Cells
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metabolism
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Plant Development
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Plant Proteins
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genetics
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metabolism
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Plants
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immunology
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Pollen Tube
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cytology
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growth & development
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immunology
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metabolism
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Signal Transduction
2.Preparation and verification of antibodies for five rice receptor-like kinases.
Bo WANG ; Liyun LI ; Yinghao CAO ; Ziguang LIU ; Guozhen LIU
Chinese Journal of Biotechnology 2010;26(6):796-801
Receptor-like kinase involves self-incompatibility, male sterility, stress responses, and disease resistance. To better understand the physiological function and biological characteristics of rice receptor-like kinase, we cloned five predicted epitope fragments of rice receptor-like kinase. The purified fusion protein was used as antigen to immunize rabbit to get specific polyclonal antibodies. Western blotting analysis shows that the five receptor-like kinases were expressed in rice leaves.
Animals
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Antibodies, Monoclonal
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biosynthesis
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immunology
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Cloning, Molecular
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Epitopes
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immunology
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Immunization
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Oryza
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enzymology
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genetics
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Plant Proteins
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genetics
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immunology
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Plantibodies
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immunology
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metabolism
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Protein Kinases
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genetics
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immunology
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Rabbits
3.Construction of rice stripe virus NS2 and NS3 Co-RNAi transgenic rice and disease-resistance analysis.
Lu-ping ZHENG ; Chen LIN ; Li-yan XIE ; Zu-jian WU ; Lian-hui XIE
Chinese Journal of Virology 2014;30(6):661-667
NS2 and NS3 are two post-transcriptional gene silencing suppressors that are encoded by Rice stripe virus. Gene silencing suppressors are always related to the pathogenicity of viruses. In this study, the cDNA of NS2 and NS3 were recombined by overlapping PCR assays, ligated to the RNAi vector, and inserted into the PXQ expression vector using Pst I; the expressed vector was transferred into calluses induced from seeds of the japonica rice cultivar, 'Nipponbare', using an Agrobacterium-mediated method. Thirty-one T0 transgenic plants were selected by G418 screening. PCR and southern blot analyses confirmed that the target gene was transformed into transgenic rice successfully, and different transgenic plants contained various copies of the gene. The disease resistance assay revealed that T0 transgenic rice had a delayed onset of RSV for approximately 10-20 d, and the accumulation of virus in the transgenic plants was reduced by 30%-50%. This was related to the delayed onset of disease.
Disease Resistance
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Oryza
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genetics
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immunology
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virology
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Plant Diseases
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genetics
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immunology
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virology
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Plants, Genetically Modified
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genetics
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immunology
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virology
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RNA Interference
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Tenuivirus
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genetics
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immunology
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Viral Nonstructural Proteins
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genetics
;
immunology
4.Comparison of Specific IgE Antibodies to Wheat Component Allergens in Two Phenotypes of Wheat Allergy.
Young Hee NAM ; Eui Kyung HWANG ; Hyun Jung JIN ; Jeong Min LEE ; Yoo Seob SHIN ; Young Min YE ; Arantxa PALACIN ; Gabriel SALCEDO ; Soo Young LEE ; Hae Sim PARK
Journal of Korean Medical Science 2013;28(11):1697-1699
Specific IgE to gliadin was proposed as a marker for wheat dependent exercise induced anaphylaxis, while Tri a 14 was found to induce IgE response in baker's asthma. We evaluated whether these components could be used for discriminating phenotypes of wheat allergy. Twenty-nine patients who were wheat-induced anaphylaxis and/or urticaria (n=21, group I) and baker's asthma (n=8, group II) were enrolled. The prevalence of serum specific IgE to Tri a 14 was higher in group II (25%) than in group I (4.8%), while the serum specific IgE to gliadin was significantly higher in group I (70%) than in group II (12.5%). The cutoff value for predicting the baker's asthma using the ratio of serum specific IgE to Tri a 14 to gliadin was 742.8 optical densityx1,000/(kU/L) with high sensitivity and specificity. These findings suggest that Tri a 14/gliadin may be a potential marker for predicting baker's asthma.
Adult
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Anaphylaxis/immunology
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Antigens, Plant/*immunology
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Asthma/blood/diagnosis/immunology
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Biological Markers/blood
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Carrier Proteins/*immunology
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Female
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Gliadin/*immunology
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Humans
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Immunoglobulin E/*blood/immunology
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Male
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Phenotype
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Triticum/immunology
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Urticaria/immunology
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Wheat Hypersensitivity/*diagnosis/*immunology
5.A comparison study on allergen components between Korean (Arachis fastigiata Shinpung) and American peanut (Arachis hypogaea Runner).
Chun Wook PARK ; Gyeong Il KIM ; Cheol Heon LEE
Journal of Korean Medical Science 2000;15(4):387-392
The prevalence of peanut allergy in Korea is lower than in America. Peanut extract allergens between the two countries have not been standardized. This study was performed to compare the allergenicity of raw Korean and American peanuts with that of roasted peanuts. We prepared the peanut extracts in Korean raw (KP) and roasted peanuts (KRP), and also in American raw (AP) and roasted (ARP) peanuts. We compared the peanut extract allergens of KP, KRP, AP and ARP in vitro with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by immunoblotting, T-cell proliferation assay and skin prick test with sera from peanut-allergic patients. SDS-PAGE and Western blotting demonstrated four allergenic extracts, numerous bands that displayed a high prevalence of IgE binding. IgE-binding bands were at 64, 36 and 17 kDa. Western blot inhibition revealed that either KP or AP could almost completely inhibit the reactivity of the other extract. There were no differences between T-cell proliferation assay and skin prick test. In conclusion, this investigation showed no different allergic components in both raw and roast extracts of Korean and American peanuts.
Allergens/immunology
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Allergens/analysis+ACo-
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Allergens/adverse effects
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Blotting, Western
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Cells, Cultured
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Comparative Study
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Electrophoresis, Polyacrylamide Gel
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Food Hypersensitivity/immunology
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Heat
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Human
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Hybridization
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Korea
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Lymphocyte Transformation
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North America
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Peanuts/immunology
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Peanuts/classification
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Peanuts/chemistry+ACo-
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Plant Extracts/immunology
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Plant Proteins/immunology
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Plant Proteins/analysis+ACo-
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Plant Proteins/adverse effects
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Skin Tests
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Species Specificity
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T-Lymphocytes/immunology
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T-Lymphocytes/drug effects
6.Progress of researches on the allergens Ara h 1, Ara h 2 and Ara h 3 from peanut.
Shengjuan JIANG ; Zhengyi ZHOU ; Yujun SUN ; Songhua WANG
Journal of Biomedical Engineering 2010;27(6):1401-1405
Peanut is one of the most popular foods in the world due to its high nutrition; however, it contains multiple seed storage proteins which are identified as allergens and hence are the most common cause of life-threatening, IgE-mediated anaphylaxis among the hypersensitive individuals. Three peanut proteins, Arachis hypogaea allergy 1, 2, 3 (Ara h 1, Ara h 2 and Ara h 3), which have the common biochemical characteristics like resistance to proteases and heat, are considered as the major allergens because they are recognized by serum IgE from a peanut-allergic patient population. The linear IgE-binding epitopes in the allergens lay the foundation of the anaphylaxis in the peanut-allergic individuals. Peanut allergy is often a life-long problem, so many investigators are focusing on decreasing clinical reactivity. In this review, the latest advances in the researches on biochemical characteristics, structure and function of the three major allergens were described and particular attention was given to the immunity properties of the three allergens. The future research directions were also discussed.
2S Albumins, Plant
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chemistry
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genetics
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immunology
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Animals
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Antigens, Plant
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chemistry
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genetics
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immunology
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Arachis
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chemistry
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DNA
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genetics
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Glycoproteins
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chemistry
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genetics
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immunology
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Humans
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Immunoglobulin E
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genetics
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Plant Proteins
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chemistry
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genetics
;
immunology
7.Mechanism analysis of broad-spectrum disease resistance induced by expression of anti-apoptotic p35 gene in tobacco.
Zhihua WANG ; Jianhua SONG ; Yong ZHANG ; Baoyu YANG ; Yao WANG ; Shiyun CHEN
Chinese Journal of Biotechnology 2008;24(10):1707-1713
Studies have shown that transgenic plants expressing antiapoptotic genes from baculovirus and animals increase resistance to biotic and abiotic stress. However, the mechanism under these resistances is conjectural, or in some cases even controversy. In the present study, the p35 gene from baculovirus Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) was expressed in tobacco, and for the first time P35 protein was detected in transgenic plants by Western blotting. Inoculation of T1 transgenic tobacco leaves with tobacco mosaic virus (TMV) showed enhanced resistance, and DNA laddering was observed after TMV infection in control but not in transgenic plants. DAB staining showed that TMV infection did not affect peroxide induction of transgenic plants, Western blotting analysis of PR1 protein also showed no difference of control and transgenic plants. Inoculation of fungus (Sclerotinia sclerotiorum) using a detached leaf assay showed enhanced resistance of transgenic leave tissue. RT-PCR analysis demonstrated that p35 gene expression induced earlier expression of PR1 gene after S. sclerotiorum infection. Taken together, our results suggest that the mechanism under enhanced disease resistance by P35 protein is possibly related to the activation of PR-related proteins in addition to the inhibition of programmed cell death, depending on the pathogens challenged.
Gene Expression Regulation, Plant
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Immunity, Innate
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Plant Diseases
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genetics
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Plants, Genetically Modified
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genetics
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immunology
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virology
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Tobacco
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genetics
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immunology
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virology
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Tobacco Mosaic Virus
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Transformation, Genetic
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Viral Proteins
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genetics
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metabolism
8.Pollen viability and stigma receptivity of Salvia miltiorrhiza and its relative.
Guocheng ZHONG ; Li ZHANG ; Ruiwu YANG ; Chunbang DING
China Journal of Chinese Materia Medica 2010;35(6):686-689
OBJECTIVETo provide the basal data for artificial cross breeding of Chinese herb Salvia miltiorrhiza from 7 provinces in China and its 4 relatives.
METHODThe pollen viability was evaluated by TTC (2, 3, 5-triphenylte trazolium chloride) test and the stigma receptivity was evaluated by benzidine-H2O2 method.
RESULTThe pollen viability of S. miltiorrhiza from 6 provinces in China and its 4 relatives deceased during time of pollen shedding. Their highest pollen viability was in 2 or 3 days after blooming. But the pollen viability of S. miltiorrhiza (wild and culture) from Hean province in China declined with time after blooming. The most obvious variation of the pollen viability was in S. miltiorrhiza from Shanxi province (RSD 71.3% ) and the least was in wild S. miltiorrhiza from Henan province (RSD 12.4%). The highest average pollen viability was wild S. miltiorrhiza (72.3%) from Henan province while the lowest was S. yunnanensis (38.8%). The stigmas of all the accessions had receptivity when blooming. The stigma receptivity of S. brevilabra was strong in 2 to 4 days after blooming, while the others had less change after blooming. The life span of pollen grains and stigmas could be maintained from 3 to 5 days.
CONCLUSIONThe optimum artificial pollination time of S. miltiorrhiza and its relatives was 2 to 3 days after blooming.
China ; Christianity ; Chromosomes, Plant ; physiology ; DNA, Plant ; analysis ; Flowers ; growth & development ; physiology ; Genetic Variation ; Genetics, Population ; Hydrogen Peroxide ; pharmacology ; Plant Infertility ; physiology ; Plant Proteins ; genetics ; Pollen ; Pollination ; immunology ; physiology ; Polyploidy ; Salvia miltiorrhiza ; physiology
9.Cloning, expression, purification of spinach carboxyl-terminal processing protease of D1 protein with hydrolysis activity and preparation of polyclonal antibody.
Hui LI ; Wei ZHANG ; Mingxia SHENG ; Weiguo LI ; Yanli LIU ; Sufang LIU ; Chao QI
Chinese Journal of Biotechnology 2010;26(4):495-502
Carboxyl-terminal processing protease of D1 protein (CtpA) catalyzes carboxyl terminal processing of the D1 protein of photosystem II, which is essential for the assembly of a manganese cluster and consequent light-mediated water oxidation. It is a target for the discovery of wide-spectrum herbicide. We amplified the CtpA gene from spinach cDNA with standard PCR method and constructed it into pET-28a vector to generate a recombinant expression plasmid. Recombinant CtpA fusion protein with His-tag was expressed as soluble protein in Escherichia coli BL21(DE3) after induction with 0.1 mmol/L IPTG at 8 degrees C for 72 h. We purified the CtpA protein with the Ni-NTA affinity chromatography and Superdex 75 gel filtration chromatography respectively, and verified the protein by SDS-PAGE and Western blotting with anti-his antibody. Hydrolysis activity of CtpA was assayed by HPLC method with a synthetic 24-mer oligopeptide corresponding to carboxyl terminal of precursor D1 protein, and gave a total activity of 1.10 nmol/(mg x min). We used the purified CtpA protein as antigen to immune rabbit for the production of polyclonal antibody, and prepared antibody with high specificity and sensitivity. The results obtained in this paper provided the feasibility of high-throughput screening of lead compounds for the protease as inhibitors and mechanism analysis of CtpA enzyme.
Algal Proteins
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Antibodies
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metabolism
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Carboxypeptidases
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biosynthesis
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chemistry
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genetics
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immunology
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Cloning, Molecular
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DNA, Complementary
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genetics
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Escherichia coli
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genetics
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metabolism
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Hydrolysis
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Proprotein Convertases
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biosynthesis
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chemistry
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genetics
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immunology
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RNA, Plant
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genetics
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Recombinant Fusion Proteins
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biosynthesis
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genetics
;
immunology
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Spinacia oleracea
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enzymology
;
genetics
10.Regulating effects of Rosa roxburghii tratt preparation on immune function in arseniasis patients caused by coal burning.
Jun LI ; Ai-hua ZHANG ; Yu-jiang REN ; Zhong-yi LIU ; Xiao-xin HUANG ; Da-ping YANG
Chinese Journal of Preventive Medicine 2013;47(9):783-787
OBJECTIVETo explore the influence of rosa roxburghii tratt preparation on immune function of arseniasis patients caused by burning coal.
METHODSAccording to the diagnosis standard for endemic arseniasis(WS/T 211-2001), 62 cases of arseniasis patients who resided in endemic arseniasis area in Guizhou province were selected and divided stratified randomly into two groups: rosa roxburghii tratt juice treatment group and superoxide dismutase(SOD)-enriched rosa roxburghii tratt juice treatment group, with 31 patients in each group.Each patient took 120 ml/d rosa roxburghii tratt preparation or SOD-enriched rosa roxburghii tratt orally for one month. Another 30 healthy residents from a neighbour township 12 km away where arsenic was not prevalent were selected as controls. A 2 ml blood and 50 ml urine samples were collected from individuals and the urine arsenic contents, peripheral blood T-lymphocyte subsets (CD3(+), CD4(+), CD8(+) T cell), serum immunoglobulin (IgG, IgM, IgA) and complement (C3, C4) were detected. The differences between more than two groups on above indicators were compared. The correlations between urinary arsenic and immune parameters were analyzed.
RESULTSAmong the rosa roxburghii tratt juice group, SOD-enriched rosa roxburghii tratt juice before intervention group and the control group, the levels of urine arsenic were (76.55 ± 23.02) , (72.60 ± 25.91) and (26.33 ± 11.30) µg/g Cr respectively and IgG were (11.31 ± 1.68), (11.35 ± 1.94) and (9.23 ± 1.75) g/L respectively. The differences were statistically significant(F values were 82.01, 13.82, both P values < 0.05). After intervention with rosa roxburghii tratt preparation, the levels of urine arsenic were (53.21 ± 16.51) and (51.72 ± 17.70)µg/g Cr, both decreased than before intervention (t values were 5.80 and 3.78, both P values < 0.05). The levels of CD3(+) were (44.47 ± 7.14)%, (43.44 ± 6.61)% and (70.78 ± 5.26)%, CD4(+) were (29.87 ± 5.67)%, (29.42 ± 5.87)% and (46.08 ± 5.87)%, CD4(+)/CD8(+) were(1.25 ± 0.42), (1.22 ± 0.39) and (1.79 ± 0.26) and C4 were (0.13 ± 0.08), (0.13 ± 0.09) and (0.20 ± 0.11) g/L respectively among the two treatment group before intervention and the control group. The differences were significant (F values were 178.04, 76.71, 23.13 and 5.26, all P values < 0.05). After intervention, the levels of CD3(+) were (59.73 ± 7.38)% and (66.31 ± 7.57)%, CD4(+) were (34.00 ± 7.97)% and (39.11 ± 5.81)%, CD4(+)/CD8(+) were (1.41 ± 0.37) and(1.58 ± 0.26), all increased than before intervention(t values were 12.47, 25.18, 5.41, 10.47, 3.22 and 5.05, all P values < 0.05). The levels of urine arsenic and CD3(+), CD4(+), CD4(+)/CD8(+), C4 were inversely correlated correlation, while positive correlation existed between the level of urine arsenic and IgG(r values were -0.68, -0.56, -0.51, -0.43 and 0.36, all P values < 0.01).
CONCLUSIONSThe level of urinary arsenic level is closely related to immune function suppression in arseniasis patients caused by burning coal, rosa roxburghii tratt preparation can effectively improve immune function of arseniasis patients.
Adult ; Arsenic ; urine ; Arsenic Poisoning ; etiology ; immunology ; China ; Coal ; Complement System Proteins ; immunology ; Female ; Humans ; Immunoglobulins ; immunology ; Male ; Middle Aged ; Plant Extracts ; pharmacology ; Rosa ; chemistry ; Superoxide Dismutase ; pharmacology ; T-Lymphocyte Subsets ; immunology