Bioregulators have a great effect on vital processes of plant growth and development. Known plant bioregulators include Naphthalene acetic acid (NAA), Indole-3-butyric acid (IBA) and Indole-3-acetic acid (IAA). Natural or synthetic plant bioregulators are organic compounds that affect the physiological processes in the plant, either to control some of these processes or to modify them. For example these bioregulators can affect the nature of the process, either by accelerating or decelerating plant growth, rates of maturation and also by altering the behavior of the plants or their products. Also, enhancement of important nutrients in human diet could be achieved by bioregulators. This study uses the model crop plant Tomato (Lycopersicon esculentum). Tomato is affected by a group of bioregulators, this group contains compounds which are powerful antioxidants in vitro. The current study aims to find out the effect of some plant bioregulators (IAA, IBA and NAA) on tomato growth, total protein content and enzyme activities of ascorbate peroxidase (APX), superoxide dismutase (SOD) and catalase (CAT). This study also investigates the effect of the above mentioned bioregulators on the level of RNA expression for SOD, CAT and TPX1 genes. The analytical quantification of target gene expression showed the induced effect of NAA on SOD expression and reducing effect of the other bioregulators (IAA and IBA) on CAT and TPX1 expression. However, at the protein level, we foundthat IBA and IAA caused a minor effect on total protein content while a significant effect was recorded on the total protein level using NAA. Upon measuring the enzyme activity of ascorbate peroxidase and catalase, we found that both the exogenous NAA and IBA stimulated ascorbate peroxidase activity in tomato while there was no considerable difference detected in IAA treated plants. Also, there was no considerable difference detected in catalase activity of all bioregulator-treated plants compared with the control.
Two different Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae) nucleopolyhedrovirus (SpliNPV) isolates were obtained from natural infected S. littoralis larvae in Egypt. The phylogenetic analysis of the partial nucleotide sequence of the polyhedrin gene (polh) showed that both isolates, Spli-6 and Spli-7, were closed and had a common ancestor with S. littoralis NPV isolate 1263 polyhedrin gene, partial coding sequence with accession number AGE90003.1 that belongs to group II NPVs. This indicates that the natural host of Spli-6 and Spli-7 must be S. littoralis. The virulence of these isolates was tested against S. littoralis larvae in a laboratory. The LC 50 against 1st instar larvae was significantly different, 3 × 10 4 OBs/ml and 9.5 × 10 4 OBs/ml for Spli-6 and Spli-7, respectively. However, the LC 50 for the mixed infection of both isolates was 3.9 × 10 4 OBs/ml. On the other hand, the ST 50 was 96 h for both isolates, while it was 120 h for the mixed infection, which was higher than that observed of each single isolate. Upon digestion of viral DNA with ScaI endonuclease, the restriction profile showed one more fragment of about~25 kbp and 3 fragments of about~19,~5.8, and~5.3 kbp in Spli-7 isolate compared to Spli-6 as well as the reference strain SpliNPV-AN1956. The differences in the fragment size and number suggest the existence of genotypic variants between both isolates. Spli-6 and Spli-7 proved to possess promising insecticidal properties for the development of virus-based biopesticide for the control of S. littoralis.
DNA encoding the coat protein (CP) of an Egyptian isolate of tomato yellow leaf curl virus (TYLCV) was inserted into the genome of Autographa californica nucleopolyhedrovirus (AcNPV) under the control of polyhedrin promoter. The generated recombinant baculovirus construct harboring the coat protein gene was characterized using PCR analysis. The recombinant coat protein expressed in infected insect cells was used as a coating antigen in an indirect Enzyme-linked immunosorbent assay (ELISA) and dot blot to test its utility for the detection of antibody generated against TYLCV virus particles. The results of ELISA and dot blot showed that the TYLCV-antibodies reacted positively with extracts of infected cells using the recombinant virus as a coating antigen with strong signals as well as the TYLCV infected tomato and beat plant extracts as positive samples. Scanning electron microscope examination showed that the expressed TYLCV coat protein was self-assembled into virus-like particles (VLPs) similar in size and morphology to TYLCV virus particles. These results concluded that, the expressed coat protein of TYLCV using baculovirus vector system is a reliable candidate for generation of anti-CP antibody for inexpensive detection of TYLCV-infected plants using indirect CP-ELISA or dot blot with high specificity.
Foot-and-mouth disease virus (FMDV) is one of the most devastating animal viruses that affect livestock worldwide. The 1B capsid of FMDV has been widely used to detect and confirm the infection. In the present study, the sequence coding for 1B subunit of FMDV capsid was expressed in insect cells using the baculovirus expression system under the polyhedrin (polh) promoter. The expression of 1B capsid protein was validated in the culture filtrate of insect cells using SDS-PAGE and western blotting. The culture filtrate containing recombinant 1B capsid (r1B) was used as a coated antigen in an indirect enzyme-linked immunosorbent assay (ELISA). The antigenicity and specificity of r1B against SAT 2 serotype-specific antibodies were assessed. Our results revealed that a protein concentration as low as 25 ng could detect SAT 2-specific antibodies in ELISA. The results highlight the application of insect cells developed r1B protein in the detection of FMDV. Further studies are required to determine the ability of r1B to detect other FMDV serotypes.
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