Detecting Viruses By RT-PCR In Fritillaria Thunbergii And Fuling Polysaccharide On Non-specific Immune Function In Pelodiscus Sinensis By Oral Administration

Fritillaria_Thumbergii Miq is a medicinal plant. The bulb of this species was employed as medicinal usage as heat-transmission, moistening the lung, relieving a cough and so on. In the field, the yield of production largely varies each year duo to infection from a wide range of many unknown pathogens. In this study, we describe some simple and reliable methods of nucleic acid extraction and report, at first time, the detection of potato leafroll virus (PLRV) and lily mottle virus (LMoV) by the reverse transcription polymerase chain reaction (RT-PCR) in Fritillaria_Thumbergii Miq with ten synthesized pairs from the serial of plant viruses. The amplified fragments with the expected size 336 bp for PLRV and 513 bp for LMoV were obtained by RT-PCR respectively, and the bands were clear. In addition, the distribution of the two viruses in the field was also evaluated. The plant materials were random detected, the infective rations are 40% and 12.5% respectively. We have detected seedling from tissue culture, the results suggested tissue culture can effectively wipe off PLRV and LMoV.Protein patterns were observed between infected bulbous tissue and uninfected bulbous tissue by SDS-PAGE gel electrophoresis and two-dimensional gel electrophoresis. SDS-PAGE gel electrophoresis shown there were two new distinct bands in infected tissues, about 35.0 kDa and 25.0 kDa. Another, at the same three protein bands, the abundance in uninfected tissues was higher than in infected tissues. Protein spots were analyzed with PDQuest software. The results show that there are large differences in protein expression level between infected and uninfected tissues. There were six new protein spots in infected tissues, but four protein spots were expressed in uninfected tissues. Compared to the healthy tissues, seven protein spots were significant boosted in expressed quantity in infected tissues, but ten spots were decline.The effect of the dietary ruling polysaccharide on non-specific immune function in soft-shelled turtle (Pelodiscus sinensis) was studied. The turtles were divided into 3 groups and were fed with feed supplemented with ruling polysaccharide and immunopolysaccharide (yeast cell wall), that was 20.0 mg/kg (fuling polysaccharide group), 500.0 mg/kg.B.W.(immunopolysaccharide group)and 0.0 mg/kg.B.W. (control group), respectively. 28 days later, the turtles were blooded, and the phagocytic activity of the leucocytes, serum lysozyme activity and complement activity were monitored. The results showed that the phagocytic activity and lysozyme activity in ruling polysaccharide group and immunopolysaccharide group were significantly higher than the control group (t-test, p<0.05). However, there were no significant differences between fuling polysaccharide group and immunopolysaccharide group (t-test, p>0.05). The highest complement activity were observed in fuling polysaccharide group and immunopolysaccharide group, which were significantly higher than the control group (t-test, p<0.05). As the complement activity was concerned, there were no significant...