Molecular cloning of vitellogenin cDNA in rockfish (Sebastes schlegeli) and effects of estrogenic substances on its gene expression

In this study, we have cloned a partial cDNA of 620 bases encoding a vitellogenin gene of rockfish (rfVTG), Sebastes schlegeli. The expression levels of rfVTG mRNA increased 48 h after the 4-nonylphenol (NP) treatment (10 mg/kg BW) in both sexes. On the other hand, the mRNA levels of rfVTG increased 48 h in males and 12 h in females after injection when higher dose (25 mg/kg BW) of 4-NP was applied. It suggested that 4-NP induced VTG gene expression in both male and female immature rockfish. Therefore, the cloned rfVTG cDNA in this study can be used as a tool for monitoring endocrine disruptors in rockfish.     

Comparison and Analysis of QTLs,Epistatic Effects and QTL×Environment Interactions for Yield Traits Using DH and RILs Populations in Rice

Two genetic linkage maps, constructed by DH and RILs populations derived from the same parents, were carried out for the identification and comparison of QTLs controlling yield traits across different years in rice (Oryza sativa L.). A total of 194 SSR and STS markers were used in two maps, of which 114 markers were same. The distribution of Samgang allele was higher in RILs population than it in DH population. Comparing with DH population, RILs population has more lines with higher yield and wider phenotypic transgressive segression for yield traits. Although most of QTLs for the same trait were different in two populations across different years, 8 QTLs (including gwp11.1, spp5.1, spp10.1, spp11.2, ssr1.1, ssr11.1, tgw9.1 and tgw11.1) were detected over 2 yr. It is important to note that ppp10.1, spp10.1 and tgw9.1 were identified in two populations, while spp10.1 and tgw9.1 were simultaneity observed across different years. Epistatic effects were more important than additive effects for PPP, SPP, yield in DH population and TGW, yield in RILs population. Epistatic effects of DH and RILs populations were different on the same genetic background in the present study, which illuminated the QE interaction played an important role on epistatic effect. Identification and comparison of QTLs for yield traits in DH and RILs populations should provide various and more precise information. The QTLs identified in present study would be valuable in marker-assisted selection program for improving rice yield.     

Freezing and Frost Damage of Potato Plants: a Case Study on Growth Recovery,Yield Response,and Quality Changes

The relationship between the severity of natural freezing and frost damage (FFD) and the observed growth recovery and field production of Superior potatoes (Solanum tuberosum L.) was investigated under screen house conditions. Potato plants were damaged by accidental freezing and frost during the early phase of shoot growth, 40 days after planting in highland summer crop areas in Korea. The observations of FFD were classified visually into “severe” (>75%), “moderate” (50%), “mild” (<25%), and “no FFD”, based on the percentage of the area of the shoots which was damaged. The early vegetative growth recovery, in terms of groundcover, was reduced as the level of FFD increased. At 28 days after a freezing and frost damage (DAFF), the potato plants with mild or no symptoms had produced more groundcover than the plants with moderate or severe symptoms. At 35 DAFF, groundcover was the same across all levels of severity, at nearly 100%. Despite the slower canopy development, plants with FFD achieved a higher fresh shoot weight and total chlorophyll content, whereas physiological growth indices such as photosynthetic rate and stomatal conductance of leaves were not different among levels of FFD at 60 DAFF. The number of lateral stems and days to tuberization increased significantly as the level of FFD increased; however, the harvest index, the number of tubers per plant, and seed tuber production were all reduced in plants with severe FFD. In addition, the tubers from plants with severe FFD had an increased ratio of length to width and 40% more tuber eyes than tubers from undamaged plants. The elongated tubers also showed an increase in cell division, demonstrated by higher numbers of cells in the cortical zones. The aforementioned measurements were obtained from natural event and led to a deduction that Superior has a capacity to recover from FFD, unless the plants were severely damaged (>75%) early in the season.     

Quantification of Odours from Piggery Effluent Ponds using an Electronic Nose and an Artificial Neural Network

An Artificial Neural Network (ANN) and an electronic nose, AromaScan, were used to predict the piggery odour concentrations emanating from an effluent pond and to develop a confident, rapid, and cost-effective technique for odour measurement. Odour samples from five different piggery effluent ponds were analysed using the AromaScan and dynamic dilution olfactometry. The resulting sensor data were used to train the artificial neural network to correlate the responses to the odour concentrations measured by olfactometry. Effectiveness was evaluated through simulation with various pre-processing techniques and network architectures. The simulation results have shown that a two-layer back-propagation neural network, which has a tan-sigmoid transfer function in the hidden layer and a linear transfer function in the output layer, could be trained to predict piggery odour concentrations with high value of the correlation coefficient R of 0·984 under the best network performance. The results from the application of scaling and principal component analysis suggest that these techniques are necessary not only to avoid the failure of the network caused by saturation but also to enhance performance. An early stopping technique was shown to provide benefits to the network performance in terms of a decrease in computation time and overfitting. It was found that the optimal number of hidden neurons for the network was 20. Odour concentration of unknown samples were able to be predicted with significant accuracy.     

Preparation of poly(vinyl acetate)/clay and poly(vinyl acetate)/poly(vinyl alcohol)/clay microspheres

Poly(vinyl acetate) (PVAc)/poly(vinyl alcohol)(PVA)/montmorillonite (MMT) clay nanocomposite microspheres with a core/shell structure have been developed via a suspension polymerization approach. In order to prepare the PVAc/MMT and PVAc/PVA/MMT nanocomposite microspheres, which are promising precursor of PVA/MMT nanocomposite microspheres, suspension polymerization of vinyl acetate with organophilic MMT and heterogeneous saponification were conducted. A quaternary ammonium salt, cetyltrimethylammonium bromide, was mixed with the MMT in the monomer phase prior to the suspension polymerization. The rate of conversion decreased with an increase in MMT concentration. The incorporation of MMT into the PVAc was verified by FT-IR spectroscopy. Organic vinyl acetate monomers were intercalated into the interlayer regions of organophilic clay hosts and followed by suspension polymerization. Partially saponified PVA/MMT nanocomposite microspheres with a core/shell structure were successfully prepared by heterogeneous saponification.     

Comparison and Analysis of QTLs,Epistatic Effects and QTLx Environment Interactions for Yield Traits Using DH and RILs Populations in Rice

Two genetic linkage maps, constructed by DH and RILs populations derived from the same parents, were carried out for the identification and comparison of QTLs controlling yield traits across different years in rice (Oryza sativa L.). A total of 194 SSR and STS markers were used in two maps, of which 114 markers were same. The distribution of Samgang allele was higher in RILs population than it in DH population. Comparing with DH population, RILs population has more lines with higher yield and wider phenotypic transgressive segression for yield traits. Although most of QTLs for the same trait were different in two populations across different years, 8 QTLs (including gwp11.1, spp5.1, spp10.1, spp11.2, ssr1.1, ssr11.1, tgw9.1 and tgw11.1) were detected over 2 yr. It is important to note that ppp10.1, spp10.1 and tgw9.1 were identified in two populations, while spp10.1 and tgw9.1 were simultaneity observed across different years. Epistatic effects were more important than additive effects for PPP, SPP, yield in DH population and TGW, yield in RILs population. Epistatic effects of DH and RILs populations were different on the same genetic background in the present study, which illuminated the QE interaction played an important role on epistatic effect. Identification and comparison of QTLs for yield traits in DH and RILs populations should provide various and more precise information. The QTLs identified in present study would be valuable in marker-assisted selection program for improving rice yield.     

Production and characterization of monoclonal antibodies detecting chicken interleukin-2 and the development of an antigen capture enzyme-linked immunosorbent assay

Eleven monoclonal antibodies (mAbs) which are specific for chicken interleukin-2 (chIL-2) were produced and characterized by enzyme-linked immunosorbent assay (ELISA), Western blotting and neutralizing assays. These mAbs were used to develop a mAb-based antigen capture ELISA specific for chicken IL-2 detection. Anti-IL-2 mAbs bound specifically to E. coli-derived rchIL-2 in ELISA and identified a 16 kDa IL-2 polypeptide band in Western blot. Several mAbs were shown to neutralize the biological activities of both rchIL-2 and native chicken IL-2 as measured by concanavalin A (ConA)-induced lymphocyte proliferation assay, IL-2 bioassay, and natural killer cell assay. Among the neutralizing mAbs, the mAb chIL-2/11 was most potent in neutralizing IL-2 activity. To develop a sensitive ELISA for the detection of chicken IL-2, an antigen capture ELISA was developed using the mAb chIL-2/16 as the antigen capture antibody and rabbit anti-IL-2 peptide antibody as the detection antibody. Using the mAb-based antigen capture ELISA, significant correlation between the level of IL-2 detected in bioassays and in ELISA was observed. These results showed that the mAb-based antigen capture ELISA is less time-consuming and more reliable compared to a conventional IL-2 bioassay for chicken IL-2. These neutralizing mAbs will facilitate basic immunobiological studies of the role of IL-2 in normal and disease states in chickens.