A flow-through immunoassay (FTA), an improved version of immunodot, was developed using a nitrocellulose membrane baked onto adsorbent pads enclosed in a plastic cassette to detect white spot syndrome virus (WSSV) in shrimp. Sharp purple dots developed with WSSV against the white background of the nitrocellulose membrane. The detection limits of WSSV by the FTA and immunodot were 0.312 and 1.2 μg mL(-1) crude WSSV protein, respectively. The FTA could be completed in 8-10 min compared with 90 min for immunodot. The FTA was 100 times more sensitive than 1-step polymerase chain reaction (PCR) and in between that of the 1- and 2-step PCR protocol recommended by the Office of International Epizootics (OIE). In experimental, orally infected shrimp post-larvae, WSSV was first detected 14, 16 and 18 h post-infection (hpi) by FTA, immunodot and one-step PCR, respectively. The FTA detected WSSV 2 and 4 h earlier than immunodot and one-step PCR, respectively. The FTA was more sensitive (25/27) than one-step PCR (23/27) and immunodot (23/27) for the detection of WSSV from white spot disease outbreak ponds. The reagent components of the FTA were stable giving expected results for 6 m at 4-8 °C. The FTA is available as a rapid test kit called 'RapiDot' for the early detection of WSSV under field conditions.

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Hypergravity—an evolutionarily novel environment has been exploited to comprehend the response of living organisms including plants in the context of extra-terrestrial applications. Recently, researchers have shown that hypergravity induces desired phenotypic variability in seedlings. In the present study, we tested the utility of hypergravity as a novel tool in inducing reliable phenotype/s for potential terrestrial crop improvement applications. To investigate, bread wheat seeds (UAS-375 genotype) were subjected to hypergravity treatment (10×g for 12, and 24 h), and evaluated for seedling vigor and plant growth parameters in both laboratory and greenhouse conditions. It was also attempted to elucidate the associated biochemical and hormonal changes at different stages of vegetative growth. Resultant data revealed that hypergravity treatment (10×g for 12 h) significantly enhanced root length, root volume, and root biomass in response to hypergravity. The robust seedling growth phenotype may be attributed to increased alpha-amylase and TDH enzyme activities observed in seeds treated with hypergravity. Elevated total chlorophyll content and Rubisco (55 kDa) protein expression across different stages of vegetative growth in response to hypergravity may impart physiological benefits to wheat growth. Further, hypergravity elicited robust endogenous phytohormones dynamics in root signifying altered phenotype/s. Collectively, this study for the first time describes the utility of hypergravity as a novel tool in inducing reliable root phenotype that could be potentially exploited for improving wheat varieties for better water usage management.

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The outstanding capability of two-dimensional gel electrophoresis in separating all types of proteins basically depends on the efficiency of sample preparation. Sample preparation is one of the most critical steps in two-dimensional gel electrophoresis. Unfortunately, due to severe solubility, resolution of protein on gel is usually hampered, and thus, analysis remains a difficult task. However, technically several problems are generally encountered during protein extraction and isoelectric focusing. In the present investigation, we emphasized on evaluation and comparison of six different protein solubilization methods intended for resolving and analyzing silkworm hemolymph proteins by two-dimensional gel electrophoresis. Our findings revealed that the buffer composition of 8 M urea, 4 % 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate, 40 mM Tris base, 65 mM dithiothreitol, and 0.2 % ampholyte can effectively solubilize and yields maximum protein spots.