Artificial infection by transplantation of juvenile Fasciola worms into the abdominal cavity of rats

Rats were successfully infected with Japanese Fasciola sp. by transplantation of juvenile worms (JW) or metacercariae (MC) into the abdominal cavity. Moreover, the rat was investigated on its suitability for different experiments with liver flukes. JWs or MCs transplanted intraperitoneally (IP) matured in the bile duct of rats. Moreover, more stable infections were established by inoculation of JWs than MCs. About 3 of 10-15 JWs transplanted into the abdominal cavity of a rat matured and laid eggs in the bile duct. The mean prepatent period was 63.5 days in the JW inoculated group. EPG values were kept constant at a level of 10(2)-10(3) about 100 to 230 days after the transplantation of JWs. The life span of Japanese liver flukes was estimated to be about 400 days in rats. From these results, it was concluded that the rat is suitable for various experiments with Fasciola sp.     

Molecular characterization of the CP gene and 3'UTR of Chilli veinal mottle virus from South and Southeast Asia

Twenty-four isolates of Chilli veinal mottle virus (ChiVMV) from China, India, Indonesia, Taiwan and Thailand were analysed to determine their genetic relatedness. Pathogenicity of virus isolates was confirmed by induction of systemic mosaic and/or necrotic ringspot symptoms on Capsicum annuum after mechanical inoculation. The 3' terminal sequences of the viral genomic RNA were determined. The coat protein (CP) coding regions ranged from 858 to 864 nucleotides and the 3' untranslated regions (3'UTR) from 275 to 289 nucleotides in length. All isolates had the inverted repeat sequence GUGGNNNCCAC in the 3'UTR. The DAG motif, conserved in aphid-transmitted potyviruses, was observed in all isolates. All 24 isolates were considered as belonging to ChiVMV because of their high CP amino acid and nucleotide identity (more than 94·8 and 89·5%, respectively) with the reported ChiVMV isolates including the pepper vein banding virus (PVBV), the chilli vein-banding mottle virus (CVbMV) and the CVbMV Chiengmai isolate (CVbMV-CM1). Based on phylogenetic analysis, ChiVMV isolates including all 24 isolates tested, PVBV, CVbMV and CVbMV-CM1 can be classified into three groups. In addition, a conserved region of 204 amino acids with more than 90·2% identity was identified in the C terminal of the CP gene of ChiVMV and Pepper veinal mottle virus (PVMV), and may explain the serological cross reaction between these two viruses. The conserved region may also provide useful information for developing transgenic resistance to both ChiVMV and PVMV.     

Mechanism and characteristics of protein release from lactitol-based cross-linked hydrogel

Lactitol-based cross-linked hydrogel was synthesized, and model proteins (alpha-chymotrypsin, beta-lactoglobulin, bovine serum albumin (BSA), and gamma-globulin) were incorporated into the cross-linked hydrogel. The larger-molecular-weight proteins have lower diffusivity (D(e)) in the hydrogel. Increasing temperature accelerated the diffusion rate of proteins; however, the diffusion did not follow the Arrhenius equation at temperatures above 37 degrees C. The swelling ratio of the hydrogel was slightly decreased after heating for 2 h at 37 and 45 degrees C, and significantly reduced after 1 h at 60 degrees C. Therefore, diffusion of beta-lactoglobulin and BSA may be decreased by hydrogel shrinking at temperature over 37 degrees C. The model proteins have high affinities to buffer solution compared to the hydrogel network structure, resulting in high partition coefficients (K > 1) which do not affect the calculation of D(e) values. Incorporated protein release follows the theory of hindered diffusion.     

Sheath-blight resistance QTLS in japonica rice germplasm

Sheath blight (SB), caused by Rhizoctonia solani, is a serious disease of cultivated rice (Oryza sativa L.) for which genetic resistance is in demand by breeders. With the goal of resistance (SBR)-QTL discovery in U. S. japonica breeding material, 197 doubled-haploid lines from a cross between MCR10277 (resistant) and Cocodrie (susceptible) were evaluated in field and greenhouse assays with U. S. and Colombian pathogen isolates and genotyped at 111 microsatellite marker loci. Four SBR QTLs from MCR10277 were identified, together accounting for 47% of field genetic variation. In all trials the strongest effect was provided by a chromosome-9 QTL, qsbr_9.1, but some QTLs differed for U. S. and Colombian R. solani isolates. SBR QTLs coincided with only two of several height or heading-time QTLs, suggesting that the relationship between these developmental traits and SBR is not simple. For the U. S. isolates, a microchamber greenhouse assay revealed the same QTLs as did field inoculation.