The subcellular localization and the resistance to fungal pathogen Gibberella fujikuroi of the protein encoded by Arabidopsis AtELHYPRP2 (EARLI1-LIKE HYBRID PROLINE-RICH PROTEIN 2, AT4G12500) were investigated using transgenic tobacco plants. The coding sequence of AtELHYPRP2 was amplified from genomic DNA of Col-0 ecotype. After restriction digestion, the PCR fragment was ligated into pCAMBIA1302 to produce a fusion expression vector, pCAMBIA1302-AtELHYPRP2-GFP. Then the recombinant plasmid was introduced into Agrobacterium tumefaciens strain LBA4404 and transgenic tobacco plants were regenerated and selected via leaf disc transformation method. RT-PCR and Western blotting analyses showed that AtELHYPRP2 expressed effectively in transgenic tobacco plants. Observation under laser confocal microscopy revealed that the green fluorescence of AtELHYPRP2-GFP fusion protein could overlap with the red fluorescence came from propidium iodide staining, indicating AtELHYPRP2 is localized to cell surface. Antimicrobial experiments exhibited that the constitutive expression of AtELHYPRP2 could enhance the resistance of tobacco to fungal pathogen G. fujikuroi and the infection sites could accumulate H2O2 obviously. The basal expression levels of PR1 and the systemic expression levels of PR1 and PR5 in transgenic tobacco plants were higher than that of the wild-type plants, suggesting AtELHYPRP2 may play a role in systemic acquired resistance.
Agrobacterium tumefaciens ; Arabidopsis ; Arabidopsis Proteins ; genetics ; Disease Resistance ; Gibberella ; pathogenicity ; Hydrogen Peroxide ; Plants, Genetically Modified ; microbiology ; Recombinant Fusion Proteins ; genetics ; Tobacco ; genetics ; microbiology

Objective To study the toxicity of fosthiazate feeding for 90 days in rats, and to determine the maximal non-effective dose of fosthiazate , in order to provide the reference dose for safety in production and chronic toxicity experiment.Methods A total of 80 SD rats ( half female and half male ) were randomly divided into 4 groups, respectively:0.8 mg/kg· bw· d group, 4.0 mg/kg· bw· d group, 20.0 mg/kg· bw· d group, and normal control group .The rats were sacrificed to determine the indices including serum biochemical parameters , body weight , routine urine test and organ coefficients after the end of the experiment , and the results were statistically analyzed .Results In the high dose group, the body weight gain was slowed in male and female rats .The TG and CHE in the high dose group of male rats and the TP, ALB, CREA, GLU, and CHE in the high dose group of female rats were significantly lower than those of normal control group.The ALP in the high dose group of female rats was higher than that of the normal control group .The positive rates of BIL, SG, and PRO in both male and female rats had significant differences compared with those of normal control group.The organ coefficients of brain , lung, kidney, adrenal, and testis of male rats, and the organ coefficients of brain , lung, and kidney of female rats in the high dose group were significantly higher than those of the normal control group .The ovaries and uterus in the female rats of high dose group were significantly lower than those of normal control group ( P<0.05,P<0.01).Conclusions The oral dose of fosthiazate at 4.0 mg/kg· bw· d fed for 90 days and above cause toxic effects on rats , and its maximal non-effect dose of long-term intake of low-dose fosthiazate on rats is 4 mg/kg· bw· d.

0.05).2.5 mg/kg and 5.0 mg/kg avermetine groups can increased the activity of LDH,GS,CK in cerebrum,and decreased the activity of CaN in cerebrum,compared with control group(P0.05).Conclusion Avermectine may cause the toxic effect in rat nervous system through creating the nerve metabolism enzyme activity.

Objective To develop a convenient, rapid and specific method using real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) for detection of facioscapulohumeral muscular dystrophy(FSHD). Methods Genomic DNA was extracted and digested by restricted endonuclease EcoR Ⅰ , followed by agarose electrophoresis. The DNA (< 38 kb) was retrieved from agarose electrophoretic gels. The primers and probe were designed in D4ZA gene in chromosome 4. One hundred and fifteen subjects were examined by FQ-PCR using the retrieved DNA (<38 kb) as a template and the result was analyzed by fluorescent curve comparing with positive control. Results The results by FQ-PCR showed that 13 cases were positive in 16 FSHD cases whose EcoR Ⅰ fragment sizes were known, 75 cases were negative in 78 cases of normal controls, 15 cases were positive in 16 FSHD cases diagnosed clinically whose EcoR Ⅰ fragment sizes were unknown, and 3 cases were positive in 5 cases of relatives of FSHD patients. Consistency was checked using Kappa index between the 2 gene diagnostic tests for FSHD (FQ-PCR test and the traditional Southern blotting test), and between the 2 diagnostic criterions (gene diagnosis by FQ-PCR and clinical diagnosis). The results were statistically significant (κ = 0. 765, P = 0. 002 ; κ = 0. 844, P = 0. 000). Conclusions A new genetic diagnostic method of FSHD by FQ-PCR was developed, which was more simplified and reliable compared to the time-consuming, radioactive Southern blotting. It could also detect the D4Z4 arrays in cases having deletion of p13E-11 as well as the interchromosomal exchange between 4q35 and 10q26. The new method of FQ-PCR for FSHD may be extended to utilize clinically in future.