Molecular Characterization and Phylogenetic Analysis of MADS-Box Gene VroAGL11 Associated with Stenospermocarpic Seedlessness in Muscadine Grapes.

Reduced expression of MADS-box gene AGAMOUS-LIKE11 (VviAGL11) is responsible for stenospermocarpic seedlessness in bunch grapes. This study is aimed to characterize the VviAGL11 orthologous gene (VroAGL11) in native muscadine grapes (Vitis rotundifolia) at the molecular level and analyze its divergence from other plants. The VroAGL11 transcripts were found in all muscadine cultivars tested and highly expressed in berries while barely detectable in leaves. RT-PCR and sequencing of predicted ORFs from diverse grape species showed that AGL11 transcripts were conservatively spliced. The encoded VroAGL11 protein contains highly conserved MADS-MEF2-like domain, MADS domain, K box, putative phosphorylation site and two sumoylation motifs. The muscadine VroAGL11 proteins are almost identical (99%) to that of seeded bunch cultivar, Chardonnay, except in one amino acid (A79G), but differs from mutant protein of seedless bunch grape, Sultanina, in two amino acids, R197L and T210A. Phylogenetic analysis showed that AGL11 gene of muscadine and other Vitis species formed a separate clade than that of other eudicots and monocots. Muscadine grape cultivar "Jane Bell" containing the highest percentage of seed content in berry (7.2% of berry weight) had the highest VroAGL11 expression, but almost none to nominal expression in seedless cultivars Fry Seedless (muscadine) and Reliance Seedless (bunch). These findings suggest that VroAGL11 gene controls the seed morphogenesis in muscadine grapes like in bunch grape and can be manipulated to induce stenospermocarpic seedlessness using gene editing technology.

Application of image analysis technique to determine cleaning of ohmic heating system for milk.

Cleaning of equipment is one of the major areas of concern in food industry. Image analysis technique was used to assess the cleaning effectiveness and optimize the CIP protocol for ohmic heating setup. Process parameters selected for optimization of cleaning were caustic concentration (1.0, 1.5, 2.0 and 2.5%), caustic temperature (70, 75, 80 and 85 °C), acid concentration (0.00, 0.25, 0.5 and 0.75%), and acid temperature (40, 50, 60 and 70 °C). Time for caustic treatment was varied from 5 to 20 min at an interval of 5 min, while time acid treatment was kept at a constant of 10 min. Taguchi orthogonal array design was used generate different combinations of acid and alkali concentration and temperature. Images of ohmic heating plates were taken before and after the cleaning procedure. MATLAB program was developed to analyze and extract Gray-Level Co-occurrence (GLCM) matrix properties from the image. Optimized combination was selected based on the highest value of desirability factor among all the experimental set of trials. Treatment with 1.5% caustic concentration at 70 °C for 5 min followed by 0.5% nitric acid concentration at 60 °C was found optimum effective CIP of the heating plates used in ohmic heating setup. GLCM properties correlation, cluster prominence, cluster shade, entropy, homogeneity and inverse difference moment normalized were found suitable for analysis of cleaning effectiveness and optimization of the CIP protocol.

Stimulation of non-specific immunity, gene expression, and disease resistance in Nile Tilapia, Oreochromis niloticus (Linnaeus, 1758), by the methanolic extract of the marine macroalga, Caulerpa scalpelliformis.

AIM: The objective of the present study was to test the immunostimulating potential of marine macroalga, Caulerpa scalpelliformis, in terms of non-specific immune responses, gene expression, and disease resistance of Nile tilapia, Oreochromis niloticus (Linnaeus, 1758). MATERIALS AND METHODS: O. niloticus was injected intraperitoneally with three different doses of methanol extract of C. scalpelliformis (CSME) (2 mg/kg, 20 mg/kg, or 200 mg/kg body weight), or MacroGard™ (commercial immunostimulant, positive control, and 20 mg/kg body weight), or distilled water (untreated control). In one set of fish, 5 days post-injection, serum lysozyme, myeloperoxidase, and antiprotease activities were assayed. 24 h after injection, gene expression was analyzed in a separate set of fish. To another set of fish, 1 week post-administration of the products, fish were challenged with lethal dose 50 (LD50) dose of a live virulent pathogen, Aeromonas hydrophila and subsequent resistance to it was noted in terms of cumulative percent mortality. RESULTS: CSME increased serum lysozyme, myeloperoxidase, and antiprotease activities. There was an increase in the expression of lysozyme gene in the spleen of treated fish. Mid dose of CSME caused the minimum mortality of 10% (consequent relative percentage survival = 73) which is comparable to that of the positive control. CONCLUSION: CSME is considered to have the potential to be developed into an immunostimulant for finfish aquaculture.

Immunostimulatory effect of the aqueous leaf extract of Phyllanthus niruri on the specific and nonspecific immune responses of Oreochromis mossambicus Peters.

Plant derived immunostimulants are a promising alternative to chemotherapeutics and also perhaps vaccines. In the present study, we examined the immunostimulating properties of aqueous leaf extract of Phyllanthus niruri, an Indian traditional medicinal herb, on neutrophil activation and antibody response of Oreochromis mossambicus (Peters). Serial ten-fold diluted doses of P. niruri ranging from 0.002 mg to 20 mg were administered to two groups of O. mossambicus (n=8). One group of fishes was administered with sheep red blood cells and the primary and secondary antibody responses were estimated using direct haemagglutination assay. The other group of fishes was administered heat-aggregated BSA to assess the ability of plant extract to elicit neutrophil activation. Our results indicate a significant enhancement of both neutrophil activation and antibody response. Among the various doses tested, fishes administered 20 mg/kg body weight caused the maximal enhancement of both primary and secondary antibody response and 0.002 mg/kg showed higher neutrophil activation compared to that of the control group. This short study indicates that aqueous leaf extract of P. niruri has the potential to be used as an immunostimulant and after confirming its immunostimulatory properties by a battery of tests on other nonspecific and specific parameters and disease-protective property by challenging the fish with virulent fish pathogens, it can be used either as a routine feed supplement to activate the immune system of farmed fishes or as an adjuvant to enhance the efficacy of vaccines.

Comparison of experimental and ab initio HF and DFT vibrational spectra of benzimidazole.

In this work, we will report a combined experimental and theoretical study on molecular and vibrational structure of benzimidazole. The laser Raman and Fourier transform infrared spectra of benzimidazole were recorded in the solid phase. The equilibrium geometry, harmonic vibrational frequencies, infrared intensities, Raman scattering activities, depolarization ratios and reduced masses were calculated by HF and density functional B3LYP method with the 6-311G(d,p) basis set. The scaled theoretical wavenumbers showed very good agreement with the experimental values. The thermodynamic functions of the title compound were also performed at HF/6-31G(d,p)/6-311G(d,p) and B3LYP/6-31G(d,p)/6-311G(d,p) levels of theory. A detailed interpretations of the infrared and Raman spectra of benzimidazole is reported. The theoretical spectrograms for FT-IR spectra of the title molecule have been constructed.