ABSTRACT
c-Myc protein encoded by c-Myc (cellular-myelocytomatosis viral oncogene) gene regulates the related gene expression through the Wnt/β-catenin signaling pathway, and has received extensive attention in recent years. The purpose of this study was to express Helicoverpa armigera c-Myc gene (Ha-c-Myc) by using prokaryotic expression system, prepare the polyclonal antibody, examine the spatio-temporal expression profile of Ha-c-Myc, and investigate the possible function of Ha-c-Myc in regulating H. armigera sterol carrier protein-2 (SCP-2) gene expression. The Ha-c-Myc gene was amplified by PCR and cloned into a prokaryotic expression plasmid pET-32a(+). The recombinant plasmid pET-32a-Ha-c-Myc was transformed into Escherichia coli BL21. IPTG was used to induce the expression of the recombinant protein. Protein was purified by Ni2+-NTA column and used to immunize New Zealand rabbits for preparing the polyclonal antibody. The Ha-c-Myc expression levels in different developmental stages (egg, larva, prepupa, pupa, and adult) of H. armigera and different tissues (midgut, fat body, head, and epidermis) of the prepupa were determined by real-time quantitative reverse transcription PCR (qRT-PCR). Ha-c-Myc siRNA was synthesized and transfected into H. armigera Ha cells. The relative mRNA levels of Ha-c-Myc and HaSCP-2 in Ha cells were detected by qRT-PCR. Results showed that the pET-32a-Ha-c-Myc recombinant plasmid was constructed. The soluble Ha-c-Myc protein of about 65 kDa was expressed in E. coli. The polyclonal antibody was prepared. Western blotting analysis suggested that the antibody had high specificity. Enzyme linked immunosorbent assay (ELISA) showed that the titer of the antibody was high. Ha-c-Myc gene expressed at all developmental stages, with high levels in the early and late instars of larva, and the prepupal stage. Tissue expression profiles revealed that Ha-c-Myc expressed in various tissues of prepupa, with high expression level in the midgut, but low levels in the epidermis and fat body. RNAi results showed that the knockdown of Ha-c-Myc expression significantly affected transcription of HaSCP-2, leading to a 50% reduction in HaSCP-2 mRNA expression level. In conclusion, the Ha-c-Myc was expressed through a prokaryotic expression system, and the polyclonal anti-Ha-c-Myc antibody was obtained. Ha-c-Myc may promote the expression of HaSCP-2 and play an important role in the lipid metabolism of H. armigera. These results may facilitate further study on the potential role and function mechanism of Ha-c-Myc in H. armigera and provide experimental data for exploring new targets of green pesticides.
Subject(s)
Animals , Rabbits , Escherichia coli/metabolism , Enzyme-Linked Immunosorbent Assay , Moths/genetics , Blotting, Western , Larva/genetics , Isoantibodies/metabolism , Antibody SpecificityABSTRACT
The cotton bollworm, Helicoverpa armigera, is a highly polyphagous pest, causing enormous losses to variouseconomically important crops. The identification and in vitro functional validation of target genes of a pest is aprerequisite to combat pest via host-mediated RNA interference (RNAi). In the present study, six hormonalbiosynthesis genes of H. armigera were chosen and evaluated by feeding insect larvae with dsRNAs corresponding to each target gene, viz., juvenile hormone acid methyltransferase (HaJHAMT), prothoracicotropichormone (HaPTTH), pheromone biosynthesis-activating peptide (HaPBAP), molt regulating transcription factor(HaHR3), activated protein 4 (HaAP-4) and eclosion hormone precursor (HaEHP). The loss of function phenotypes for these hormonal genes were observed by releasing second instar larvae on to artificial diet containingtarget gene-specific dsRNAs. Ingestion of dsRNAs resulted in mortality ranging from 60% to 90%, reduced larvalweight, phenotypic deformities and delayed pupation. The quantitative real-time PCR (qRT-PCR) analysisshowed that the target gene transcript levels were decreased drastically (31% to 77%) as compared to control orunrelated control (GFP-dsRNA), and correlated well with the mortality and developmental defects of larvae.Also, a comparison of the silencing efficacy of un-diced long HaPTTH-dsRNA with RNase III diced HaPTTHdsRNA (siRNAs) revealed that long dsRNAs were more efficient in silencing the target gene. These resultsindicated that the hormonal biosynthesis genes have varied sensitivity towards RNAi and could be the vital targetsfor insect resistance in crop plants like cotton which are infested by H. armigera
ABSTRACT
Silkworm silk protein fibroin is widely exploited to develop novel silk-based biomaterials due to its stable b-sheetstructure, providing high crystallinity and tensile strength. The polymorphic behaviour of silk fibroin provides awindow to modulate its structural transitions during self-assembly for different functional outcomes. Most studiesare therefore mainly focused on formation of well-developed b-sheet structure and self-assembly of silk fibroinwhich are regulated by many parameters. Glyoxal, a highly reactive a-oxoaldehyde, reacts with different proteinsto form advanced glycation end products (AGEs) following Maillard-like reaction. Considering the significanceof protein modification by glyoxal-derived AGEs, in the present study the effect of glyoxal (250, 500 and1000 lM) on the structure of silk fibroin has been investigated. CD and fluorescence studies reveal that higherconcentrations of the a-oxoaldehyde induce considerable alterations of secondary and tertiary structure of theprotein leading to aggregation following incubation with for 3 weeks. The aggregates exhibit fibrillar morphologywith amyloidal nature as evident from SEM, FTIR and XRD experiments. The findings highlight that glycationinduced modification can be a possible approach for modulating the conformation of the silk protein which may berelevant in connection to clinical, biomedical or synthetic biology based applications.
ABSTRACT
Abstract Development of transgenic Bt crops with stable and high level of Bt protein expression over generations under different environmental conditions is critical for successful deployment at field level. In the present study, progenies of transgenic cotton Coker310 event, CH12 expressing novel cry2AX1 gene were evaluated in T3 generation for stable integration, expression and resistance against cotton bollworm, Helicoverpa armigera. The cry2AX1 gene showed stable inheritance and integration in the T3 progeny plants as revealed by PCR and Southern blot hybridization. The expression of Cry2AX1 protein on 90 days after sowing (DAS) was in the range of 1.055 to 1.5 µg/g of fresh leaf tissue except one plant which showed 0.806 µg/g of fresh leaf tissue and after 30 days (i.e., on 120 DAS) three plants recorded in between 0.69 to 0.82 µg/g and other plants are in range of 0.918 to 1.058 µg/g of fresh leaf tissue. Detached leaf bit bioassay in T3 progeny on 110 DAS recorded mortality of 73.33 to 93.33 per cent against H. armigera and severe growth retardation in surviving larvae. These results indicate that the expression of chimeric cry2AX1 is stable and exhibits insecticidal activity against H. armigera in T3 progeny of CH12 event of transgenic cotton.
Subject(s)
Animals , Bacillus thuringiensis/pathogenicity , Pest Control, Biological/methods , Gossypium/genetics , Endotoxins/genetics , Moths , Plant Diseases/prevention & control , Plants, Toxic , Biological Assay , Plants, Genetically ModifiedABSTRACT
Both, the tobacco caterpillar Spodoptera litura (Fabricius) and the cotton bollworm Helicoverpa armigera (Hübner), are serious polyphagous pests causing considerable loss to crops. Indiscriminate use of chemical pesticides for controlling them has rather resulted in their resistance development. Microbial pesticides, Bacillus thuringiensis in particular, play an important role in pest management. Here, we isolated Bacillus thuringiensis-like bacteria from the soil samples primarily collected from North East region of India along with some states viz., Haryana, Punjab, Maharashtra, West Bengal and Uttarakhand and studied their toxicity against the above two insect pests at 10 µg/g along with standard strain B. thuringiensis subsp. kurstaki HD-1 and at 1 µg/g Pseudomonas fluorescens based MVPII expressing Cry1Ac toxin and AUG-5. Isolates AUG-5 and GTG-7 proved toxic to more than 75% larvae on the 4th as well as 7th day of the treatment of the neonates of H. armigera. The AUG-5 isolate was also effective against S. litura. Ten effective isolates (AUG-5, GTG-4, GTG-7, GTG-9, GTG-42, GTG-64, GTG-70, GTG-3S, GTG-4S and GTG-6S) were characterized using biochemical and 16S rDNA analysis. Nearly, all the isolates tested positive for utilizing monosaccharides. All selected B. thuringiensis isolates showed resistance to ampicillin and co-trimoxazole except AUG-5 to co-trimoxazole. AUG-5 and GTG-7 were highly toxic to both insects, and possessed cry1, cry1A and cry2 genes. These isolates AUG-5 and GTG-7 also contained high Cry1Ac (104.8 and 88.32 ng/mg) and Cry2Ab (3792 and 1305.9 ng/mg), respectively in their spore-crystal complex. Both, AUG-5 and GTG-7 isolates, could be considered for further development as bioinsecticides. The present study has established the diversity and richness of B. thuringiensis-like isolates in soils collected from north-eastern region of India.
ABSTRACT
Adequate expression of Bt (Bacillus thuringiensis) toxins and purity of seeds of Bt-transgenic cottons are important for controlling bollworms, and thereby increasing the cotton productivity. Therefore, we examined the variability in expression of Bt toxin proteins in the seeds and in leaves of different cotton (Gossypium hirsutum (L.) hybrids (JKCH 226, JKCH 1947, JKCH Durga, JKCH Ishwar, JKCH Varun KDCHH 441 and KDCHH 621) expressing Bt toxins in F1 and F2 generations, using bioassays against the cotton bollworm, Helicoverpa armigera (Hübner), and the lateral flow strip (LFS) test. Toxicity of Bt toxin proteins in the seeds of Bt-transgenic cottons to H. armigera correlated with their toxicity in the leaves in one-toxin Bt cotton hybrids. The Bt-F1 and Bt-F2 seeds of JKCH 1947 were more toxic to H. armigera than those of JKCH Varun seeds. The seeds and leaves of F1s showed greater toxicity than the F2 seeds or leaves of one-toxin (cry1Ac) Bt cotton hybrids. However, no significant differences were observed for the two-toxin (cry1Ac and cry2Ab) hybrid, KDCHH 621. Toxicity of leaves to H. armigera increased with crop age, until 112 days after seedling emergence. The Bt trait purity in F1 seeds of four two-toxin Bt cotton hybrids ranged from 86.7 to 100%. The present study emphasizes the necessity of 95% Bt trait purity in seeds of transgenic cotton for sustainable crop production.
ABSTRACT
Newly developed Phytopesticidal formulations from pongam and neem oils were evaluated for their feeding deterrent activity using leaf disc choice and no-choice methods, and genotoxic study using comet assay against Helicoverpa armigera at different concentrations of 5, 10, 15, and 20 ppm. Among various phytopesticidal formulations, neem and pongam oils at 1:1 ratio, called PONNEEM showed significant feeding deterrent activity against H. armigera at 20 ppm concentration and wasgenotoxic to H. armigera (P>0.001). The comet parameters, namely tail moment (arbitrary units), tail length (µm) and tail DNA (%) were observed at all the concentrations of PONNEEM. Statistically significant changes in all the comet parameters of H. armigera were observed at 20 ppm (P<0.001). Feeding deterrent and genotoxicity effect of PONNEEM could be applied as phytopesticide for controlling the lepidopteran insect pests.
ABSTRACT
Forty-four isolates of Bacillus thuringiensis like bacteria from various sources in different locations from Sudan were tested for their insecticidal activity. The toxicity of these isolates ranged from 6.6 to 70% to the neonates of cotton bollworm, Helicoverpa armigera at 10 ppm concentration. The most effective ones are Kb-29, St-6 and Wh-1 comparable with HD-1. Toxicity of isolates to larvae of the red flour beetle, Tribolium castaneum ranged from 20 to 100%. Isolates St-2 and St-23 gave 100% larval mortality within 15 days of exposure and were at par with Ab-8, Ab-12, Kb-26, Kb-30, Om-4, Po-2, Po-5, Po-7, Sa-8 and Wh-5 and were also comparable with E. coli clone expressing Cry3 toxin. The most effective five isolates viz., Kb-29, St-2, St-6, St-23 and Wh-1 belonged to B. thuringiensis. The St-6 isolate, which also showed high toxicity to T. castaneum larvae, had cry1 genes along with coleopteran active cry28 genes, but not cry3 genes. Of the 25 isolates characterized with 16s DNA sequencing, seven belonged to Paenibacillus spp., one Lysinibacillus sphaericus, one Bacillus pumilus, four Bacillus spp., and rest 12 belonged to B. thuringiensis. Biochemical characterization in each species showed variation. The present study shows potential of some isolates like Kb-29, St-2, St-6, St-23 and Wh-1 as promising bioinsecticides.
Subject(s)
Animals , Bacillus thuringiensis/isolation & purification , Endotoxins/metabolism , Humans , Moths , Pest Control, Biological/methods , Sudan , Treatment Outcome , TriboliumABSTRACT
Objective: To evaluate a formulation from the milky mangrove tree Excoecaria agallocha L. (E. agallocha) against Helicoverpa armigera Hubner (H. armigera). Methods: About 3% aqueous ethanolic spray formulation derived from the lipophilic extract of E. agallocha (dry leaf) was evaluated against H. armigera in Abelmoschus esculentus (lady's finger) and Cajanus cajan (C. cajan) (pigeon pea), under field conditions. Results: On the 9th day of the 4th spray the larval count in the plot treated with 3% E. agallocha formulation drastically came down to 0.23 larva/plant, compared to 1.63 in the ethanol control plot and 1.60 in the unsprayed plot. Blocks sprayed with 3% E. agallocha formulation yielded 35.8 quintals/hectare (q/ha) of healthy pods compared to Ekalux® (pod yield: 60.7 q/ha), 3% Vijay Neem® (60.22 q/ha), yield plot (6 q/ha) and ethanol control (7 q/ha). In C. cajan, 1% E. agallocha, 3% Nimbecidine® and 0.07% indoxacarb were equally potent in reducing the larval population of H. armigera and the non-target pest Spilosoma obliqua to 0%, from the 9th day (3rd spray). Indoxacarb plot recorded the maximum yield of 16.1 q/ha with 2.4% pod damage. Plots sprayed with 1% E. agallocha yielded 14.7 q/ha with 2.32% pod damage. The effect of 3% Nimbecidine® spray (14.35 q/ha) was comparable to E. agallocha formulation. Unsprayed and ethanol control plots yielded 12.41 and 11.2 q/ha of pods with an average pod damage of 4.7%. Conclusions: E. agallocha formulation was found to be promising for the control of H. armigera, under field conditions.
ABSTRACT
Objective: To assess the feeding deterrent, growth inhibitory and egg hatchability effects of PONNEEM on Helicoverpa armigera (H. armigera).Methods:growth inhibitory and egg hatchability effects on H. armigera.Results:Invariably all the newly formulated phytopesticidal oil formulations showed the feeding Five oil formulations were prepared at different ratios to assess the feeding deterrent, deterrent and growth inhibitory activities against H. armigera. The maximum feeding deterrent activity of 88.44% was observed at 15 μL/L concentration of PONNEEM followed by formulation A (74.54%). PONNEEM was found to be effective in growth inhibitory activities and egg hatchability at 10 μL/L concentration. It exhibited statistically significant feeding deterrent activity and growth inhibitory activity compared with all the other treatments.Conclusions:PONNEEM was found to be effective phytopesticidal formulation to control the larval stage of H. armigera. This is the first report for the feeding deterrent activity of PONNEEM against H.armigera. This newly formulated phytopesticide was patented in India.
ABSTRACT
<p><b>OBJECTIVE</b>To assess the feeding deterrent, growth inhibitory and egg hatchability effects of PONNEEM on Helicoverpa armigera (H. armigera).</p><p><b>METHODS</b>Five oil formulations were prepared at different ratios to assess the feeding deterrent, growth inhibitory and egg hatchability effects on H. armigera.</p><p><b>RESULTS</b>Invariably all the newly formulated phytopesticidal oil formulations showed the feeding deterrent and growth inhibitory activities against H. armigera. The maximum feeding deterrent activity of 88.44% was observed at 15 µL/L concentration of PONNEEM followed by formulation A (74.54%). PONNEEM was found to be effective in growth inhibitory activities and egg hatchability at 10 µL/L concentration. It exhibited statistically significant feeding deterrent activity and growth inhibitory activity compared with all the other treatments.</p><p><b>CONCLUSIONS</b>PONNEEM was found to be effective phytopesticidal formulation to control the larval stage of H. armigera. This is the first report for the feeding deterrent activity of PONNEEM against H. armigera. This newly formulated phytopesticide was patented in India.</p>
ABSTRACT
OBJECTIVE@#To evaluate a formulation from the milky mangrove tree Excoecaria agallocha L. (E. agallocha) against Helicoverpa armigera Hubner (H. armigera).@*METHODS@#About 3% aqueous ethanolic spray formulation derived from the lipophilic extract of E. agallocha (dry leaf) was evaluated against H. armigera in Abelmoschus esculentus (lady's finger) and Cajanus cajan (C. cajan) (pigeon pea), under field conditions.@*RESULTS@#On the 9th day of the 4th spray the larval count in the plot treated with 3% E. agallocha formulation drastically came down to 0.23 larva/plant, compared to 1.63 in the ethanol control plot and 1.60 in the unsprayed plot. Blocks sprayed with 3% E. agallocha formulation yielded 35.8 quintals/hectare (q/ha) of healthy pods compared to Ekalux® (pod yield: 60.7 q/ha), 3% Vijay Neem® (60.22 q/ha), yield plot (6 q/ha) and ethanol control (7 q/ha). In C. cajan, 1% E. agallocha, 3% Nimbecidine® and 0.07% indoxacarb were equally potent in reducing the larval population of H. armigera and the non-target pest Spilosoma obliqua to 0%, from the 9th day (3rd spray). Indoxacarb plot recorded the maximum yield of 16.1 q/ha with 2.4% pod damage. Plots sprayed with 1% E. agallocha yielded 14.7 q/ha with 2.32% pod damage. The effect of 3% Nimbecidine® spray (14.35 q/ha) was comparable to E. agallocha formulation. Unsprayed and ethanol control plots yielded 12.41 and 11.2 q/ha of pods with an average pod damage of 4.7%.@*CONCLUSIONS@#E. agallocha formulation was found to be promising for the control of H. armigera, under field conditions.
ABSTRACT
This research was carried out to examine cytopathological effects of Helicoverpa armigera Cytoplasmic polyhedrosis virus (HaCPV) on infected midgut cotton bollworm (Helicoverpa armigera) using transmission and scanning electron microscope. The symptoms on infected host larvae of the host, compared with healthy ones, were getting swollen with milky-white and fragile Histopathological examinations showed infection with HaCPV small polyhedral inclusion bodies (PIB) after 1 or 2 days which were observed in columnar cells of midgut. Virions were partially or completely occupied in a polyhedral matrix to form polyhedral inclusion bodies (PIB) at periphery of virogenic stroma. PIBs were measured 0.5 to 3.5 mm and virions about 46 nm in diameter. Microvilli of infected columnar cells were affected and degenerated immediately prior to rupture of the cell. Some infected columnar cells ruptured to release PIB into the gut lumen 3 days after infection. In addition,PIB were found in goblet cells, 5 or 6 days after infection. Infected goblet cells degenerate to such an extent that only a few of the original microvillus-like cytoplasmic projections and cell organells were left. These cytopathic effects caused in the midgut by HaCPV on cotton bollworm larvae are essentially similar to those have been reported for lepidoperan and dipteran infection by CPV.
Subject(s)
Animals , Lepidoptera/virology , Reoviridae/growth & development , Gastrointestinal Tract/pathology , Histocytochemistry , Larva/virology , Microscopy, Electron, Scanning , Microscopy, Electron, TransmissionABSTRACT
Acacia nilotica proteinase inhibitor (AnPI) was isolated by ammonium sulphate precipitation followed by chromatography on DEAE-Sephadex A-25 and resulted in a purification of 10.68-fold with a 19.5% yield. Electrophoretic analysis of purified AnPI protein resolved into a single band with molecular weight of approximately 18.6+1.00 kDa. AnPI had high stability at different pH values (2.0 to 10.0) except at pH 5.0 and are thermolabile beyond 80°C for 10 min. AnPI exhibited effective against total proteolytic activity and trypsin-like activity, but did not show any inhibitory effect on chymotrypsin activity of midgut of Helicoverpa armigera. The inhibition kinetics studies against H. armigera gut trypsin are of non-competitive type. AnPI had low affinity for H. armigera gut trypsin when compared to SBTI. The partially purified and purified PI proteins-incorporated test diets showed significant reduction in mean larval and pupal weight of H. armigera. The results provide important clues in designing strategies by using the proteinase inhibitors (PIs) from the A. nilotica that can be expressed in genetically engineered plants to confer resistance to H. armigera.
ABSTRACT
Objective: To evaluate the ovicidal activity of different botanical oil formulations againstHelicoverpa armigera and Spodoptera litura. Methods: Different botanical oils were formulated with different ratio to evaluate the ovicidal activity against H. armigera and S. litura at 5, 10, 15 and 20μl/L concentrations. Results: All the oil formulations showed the ovicidal activity against H. armigera and S. litura. The maximum ovicidal actvity of 76.74 and 69.36% was noticed at 20μl/L concentration in formulation 3 PONNEEM. Formulation 4 Pongam oil showed lower ovicidal activity of 31.34 and 24.76% against H. armigera and S. litura respectively. Among the formulations, PONNEEM exhibited statistically superior ovicidal activity against both insect pests. Conclusions: the present study clearly showed PONNEEM as a pontenial biopesticide to control the egg stage of economically important pests of H. armigera and S. litura. This is the first report for the ovicidal activity of PONNEEM against these two insect pests.
ABSTRACT
Objective:To evaluate the antifeedant, insecticidal and growth inhibition activities of Solanum pseudocapsicum (S. pseudocapsicum) seed extracts against Spodoptera litura (S. litura) and Helicoverpa armigera (H. armigera). Methods:Hexane, diethyl ether, dichloromethane and ethyl acetate seed extracts were prepared and tested for antifeedant, insecticidal and growth inhibitory activities against fourth instar larvae of S. litura and H. armigera. Results:Ethyl acetate extract showed promising antifeedant and insecticidal activities against S. litura and H. armigera. Percentage of deformed larvae, pupae and adults were maximum in treatment of ethyl acetate extract. Percentage of successful adult emergence was deteriorated by seeds on extract treated larvae. Conclusions: Ethyl acetate extracts of S. pseudocapsicum, showed higher efficiency of antifeedant, insecticidal and growth inhibition activities. Hence, it can be used to controll agricultural insect pests, S. litura and H. armigera.
ABSTRACT
<p><b>OBJECTIVE</b>To evaluate the antifeedant, insecticidal and growth inhibition activities of Solanum pseudocapsicum (S. pseudocapsicum) seed extracts against Spodoptera litura (S. litura) and Helicoverpa armigera (H. armigera).</p><p><b>METHODS</b>Hexane, diethyl ether, dichloromethane and ethyl acetate seed extracts were prepared and tested for antifeedant, insecticidal and growth inhibitory activities against fourth instar larvae of S. litura and H. armigera.</p><p><b>RESULTS</b>Ethyl acetate extract showed promising antifeedant and insecticidal activities against S. litura and H. armigera. Percentage of deformed larvae, pupae and adults were maximum in treatment of ethyl acetate extract. Percentage of successful adult emergence was deteriorated by seeds on extract treated larvae.</p><p><b>CONCLUSIONS</b>Ethyl acetate extracts of S. pseudocapsicum, showed higher efficiency of antifeedant, insecticidal and growth inhibition activities. Hence, it can be used to controll agricultural insect pests, S. litura and H. armigera.</p>
Subject(s)
Animals , Humans , Insecticides , Pharmacology , Larva , Lethal Dose 50 , Moths , Pest Control , Plant Extracts , Pharmacology , Solanaceae , Chemistry , SpodopteraABSTRACT
The ichneumonid parasitoid, C. chlorideae is an important natural enemy of pod borer/bollworm, Helicoverpa armigera (Hubner) in different agro-ecosystems. The sex-ratio of parasitoids has an important bearing on the population build up of the natural enemies for biological control of insect pests. Therefore, the present studies were conducted to gain an understanding of the influence of mating behaviour and abundance of the insect host on fecundity and sex-ratio of the parasitoid, C. chlorideae. There was no significant influence of number of matings and abundance of the insect host on cocoon formation, adult emergence, and larval and pupal periods of C. chlorideae. However, fecundity and female longevity were significantly influenced by mating and abundance of the insect host. There was a significant and positive correlation (r = 0.84**) between longevity and fecundity of C. chlorideae females. The unmated C. chlorideae females produced only males. Nearly 20% of the females that had mated twice were able to parasitize the H. armigera larvae successfully. The sex-ratio of the progeny from females that had mated twice was male biased. Females mated with males from the unmated females produced significantly less numbers of females than those mated with males from the fertilized females, indicating genetic regulation of sex-ratio in C. chlorideae.
ABSTRACT
Protein phosphorylation is one of the most common post-translational modification processes that play an essential role in regulating protein functionality.The Helicoverpa armigera single nucleopolyhedrovirus (HearNPv) orf2-encoded nucleocapsid protein HA2 participates in orchestration of virus-induced actin polymerization through its WCA domain,in which phosphorylation status are supposed to be critical in respect to actin polymerization.In the present study,two putative phosphorylation sites (232Thr and 250Ser) and a highly conserved Serine (245Ser) on the WCA domain of HA2 were mutated,and their phenotypes were characterized by reintroducing the mutated HA2 into the HearNPV genome.Viral infectivity assays demonstrated that only the recombinant HearNPV bearing HA2 mutation at 245Ser can produce infectious virions,both 232Tbr and 250Ser mutations were lethal to the virus.However,actin polymerization assay demonstrated that all the three viruses bearing HA2 mutations were still capable of initiating actin polymerization in the host nucleus,which indicated the putative phosphorylation sites on HA2 may contribute to HearNPV replication through another unidentified pathway.
ABSTRACT
Baculoviruses produce two viral phenotypes, the budded virus (BV) and the occlusion-derived virus (ODV). ODVs are released from occlusion bodies in the midgut where they initiate a primary infection. Due to the lack of an in vitro system, the molecular mechanism of ODV infection is still unclear. Here we present data demonstrating that Helicoverpa armigera nucleopolyhedrovirus (HearNPV) ODV infected cultured Hz-AM1 cells in a pH dependent manner. The optimal pH for ODV infection was 8.5, which is same to that in the microvilli of midgut epithelial cells, the ODV native infection sites. Antibodies neutralization analysis indicated that four HearNPV oral infection essential genes p74, pif-1, pif-2 and pif-3 are also essential for HearNPV ODV infection in vitro. Thus, HearNPV-HzAM1 system can be used to analyze the mechanism of ODV entry.