A truncated version of the white spot syndrome virus (WSSV) 27.5 kDa envelope protein was expressed as a histidine tag fusion protein in Escherichia coli. The bacterial expression system allowed the production of up to 10 mg of purified recombinant protein per liter of bacterial culture. Antiserum from a rabbit immunized with the recombinant protein recognized the 27.5 kDa viral envelope protein of WSSV isolated from different geographic regions. The antiserum did not recognize any of the other known WSSV structural proteins. A sensitive immunodot assay for WSSV was developed using the specific rabbit polyclonal antiserum.KEY WORDS: White spot syndrome virus · 27.5 kDa structural protein · Rabbit polyclonal antiserumResale or republication not permitted without written consent of the publisher

A cDNA library was constructed from white spot syndrome virus (WSSV)-infected penaeid shrimp tissue. cDNA clones with WSSV inserts were isolated and sequenced. By comparison with DNA sequences in GenBank, cDNA clones containing sequence identical to those of the WSSV envelope protein VP28 and nucleoprotein VP15 were identified. Poly(A) sites in the mRNAs of VP28 and VP15 were identified. Genes encoding the major viral structural proteins VP28, VP26, VP24, VP19 and VP15 of 5 WSSV isolates collected from different shrimp species and/or geographical areas were sequenced and compared with those of 4 other WSSV isolate sequences in GenBank. For each of the viral structural protein genes compared, the nucleotide sequences were 100 to 99% identical among the 9 isolates. Gene probes or PCR primers based on the gene sequences of the WSSV structural proteins can be used for diagnoses and/or detection of WSSV infection.KEY WORDS: White spot syndrome virus · Viral structural proteins · VP28 · VP26 · VP24 · VP19 · VP15Resale or republication not permitted without written consent of the publisher

Rhizobium etli strain TAL182 is a competitive strain for effective nodulation of beans. From this strain, a novel gene was isolated, slp, which is 669 bp in size and required for nodulation competition on the common bean. The slp knockout mutant of TAL182 is defective in nodulation competition, shows reduced growth in the presence of 200 mM NaCl, KCl or LiCl and is complemented by the cloned slp gene. The deduced amino acid sequence of slp shows 66-72% similarity to stomatin proteins of Homo sapiens, Mus musculus and Caenorhabditis elegans. Expression of slp in Escherichia coli from a T7 promoter shows a 26 kDa protein which cross-reacts with human-stomatin-specific polyclonal antibody. Like the human stomatin protein, the slp-deduced protein, Slp, is very hydrophilic except for a single hydrophobic membrane-spanning domain. Among various bean-nodulating rhizobia, slp is present in R. etli, Rhizobium leguminosarum bv. phaseoli and Rhizobium tropici type A strains but is absent in R. tropici type B strains. It is also absent in Bradyrhizobium and several other Rhizobium spp.

Aims: To isolate and characterize bradyrhizobia that nodulate yardlong bean and sunnhemp in Guam. Methods and Results: Bradyrhizobia populations that nodulate yardlong bean and sunnhemp in Guam were examined for genetic diversity and their relatedness to Bradyrhizobium japonicum and B. elkanii reference strains. Genomic DNA of 58 isolates of Bradyrhizobium spp. was hybridized with B. japonicum nodY and B. elkanii nodK genes. Based on the hybridization patterns, the isolates were classified into three nodY-nodK hybridizing groups. Group I comprised the majority of the isolates and hybridized with nodY whereas group II isolates hybridized with nodK. The group III isolates, that did not hybridize with either nodY or nodK, formed nitrogen-fixing nodules on cowpea but did not nodulate soybean. DNA sequence analysis of a 280-bp fragment of the variable region of the 16S rRNA gene of a few group III isolates showed that these isolates were more similar to Bradyrhizobium spp. than to B. japonicum or B. elkanii. Conclusions: The majority of the isolates nodulating yardlong bean and sunnhemp in Guam are similar to B. japonicum, although some isolates are similar to Bradyrhizobium spp. that nodulate a miscellaneous group of legumes including cowpea. Significance and Impact of the Study: Since both yardlong bean and sunnhemp are nodulated by a range of bradyrhizobia, selection of superior strains may be based on nodulation effectiveness on both legumes.

Marsupenaeus japonicus has two types of phenotypic differences that are mainly reflected in the stripe pattern of the carapace. However, the underlying mechanism regulating the stripe patterns is not clear. In the present study, we first observed the composition of pigment cells and detected the contents of different carotenoids in the carapace of M. japonicus. We clearly observed the setae structure on the carapace. There were red pigment cells in the stripe pattern and yellow pigment cells in the other parts. Both red pigment cells and yellow pigment cells showed dendritic morphology. In the carapace, the content of astaxanthin was the highest, significantly (P < 0.05) higher than that of other carotenoids. Some differentially expressed genes between two pattern types of M. japonicus, may be associated with the body color formation, such as crustacyanin (CRCN), apolipoprotein D (ApoD), tubulin alpha-1 chain, cuticle protein, and ABC transporter, which were verified by quantitative PCR experiments. The amino acid composition and secondary structure of CRCN A2, CRCN C1, and ApoD were significantly different. The results of this study will help to elucidate the molecular mechanism of the differential pattern formation of M. japonicus and provide a reference for further exploration of the formation mechanism of crustacean color.