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1.
PLoS One ; 7(8): e43567, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22937062

RESUMO

BACKGROUND: The MEAM1 (B biotype) Bemisia tabaci (Gennadius) is one of the most widespread and damaging whitefly cryptic species. Our previous studies discovered that the MEAM1 whitefly indirectly benefits from interactions with the tomato yellow leaf curl China virus (TYLCCNV) via accelerated ovarian development and increased fecundity. However, the physiological mechanism of begomoviruse-infected plants acting on the reproduction of the insect vector was unknown. METHODOLOGY/PRINCIPAL FINDINGS: Biochemical and molecular properties of vitellogenin (Vg) and vitellin (Vt) were characterized in the MEAM1 whitefly. In addition, kinetics of Vt levels in ovary and Vg levels in hemolymph in different stages were detected using a sandwich ELISA. The level of hemolymph Vg increased rapidly after eclosion. A significantly higher level of hemolymph Vg and ovary Vt were observed in whiteflies feeding on virus-infected tobacco plants than those feeding on uninfected plants. In order to detect the levels of Vg mRNA transcription, complete vitellogenin (Vg) mRNA transcripts of 6474 bp were sequenced. Vg mRNA level in whiteflies feeding on virus-infected plants was higher than those feeding on uninfected plants. However, virus-infection of the whiteflies per se, as demonstrated using an artificial diet system, did not produce significant changes in Vg mRNA level. CONCLUSIONS/SIGNIFICANCE: In MEAM1 whitefly, increased levels of both vitellin and vitellogenin as well as increased transcription of Vg mRNA are associated with feeding on begomovirus-infected plants, thus providing a mechanism for accelerated vitellogenesis. We conclude that MEAM1 whitefly profits from feeding on begomovirus-infected plants for yolk protein synthesis and uptake, and thereby increases its fecundity. These results not only provide insights into the molecular and physiological mechanisms underlying the elevated reproduction of a whitefly species through its association with a begomovirus-infected plant, but also provide a better understanding of the molecular mechanisms related to whitefly reproduction.


Assuntos
Begomovirus/patogenicidade , Hemípteros/crescimento & desenvolvimento , Hemípteros/genética , Nicotiana/virologia , Animais , Feminino , Hemolinfa/metabolismo , Ovário/metabolismo , Vitelinas/genética , Vitelogênese/genética , Vitelogênese/fisiologia , Vitelogeninas/genética
2.
PLoS One ; 5(7): e11713, 2010 Jul 22.
Artigo em Inglês | MEDLINE | ID: mdl-20676356

RESUMO

BACKGROUND: Plant-pathogenic begomoviruses have a complex association with their insect vectors. The interactions of begomoviruses and reproduction of their vectors are poorly understood. Bemisia tabaci is known to transmit many begomoviruses, and the spread of B. tabaci, especially the B and Q 'biotypes', has been accompanied by the epidemics of begomoviruses. One of these identified disease-causing agents was Tomato yellow leaf curl China virus (TYLCCNV). METHODOLOGY/PRINCIPAL FINDINGS: In this study, we compared the egg production and realized fecundity of two 'biotypes' or putative species of the whitefly B. tabaci, including the alien invasive B and the indigenous ZHJ1 from Zhejiang, China, feeding on either healthy or TYLCCNV-infected tobacco plants. The ovary of the whitefly was composed of 12-22 telotrophic ovarioles. According to the morphology of the oocytes and level of yolk content, oocytes in ovarioles were divided into four developmental phases (I-IV). Significantly higher proportion of immature oocytes (phase II, III) and mature oocytes (phase IV) was observed in ovary of females that fed on TYLCCNV-infected tobacco compared to that on healthy plants. Moreover, there was significant increase of eggs laid of B whitefly that fed on TYLCCNV-infected tobacco plants during the early developmental stages. In contrast, the proportion of oocytes of different developmental phases and eggs laid had no significant differences between ZHJ1 whiteflies feeding on TYLCCNV-infected and non-infected host plants. CONCLUSIONS/SIGNIFICANCE: The invasive B whitefly benefits from feeding on a begomovirus-infected plant through increased egg production and realized fecundity.


Assuntos
Hemípteros/anatomia & histologia , Hemípteros/citologia , Nicotiana/virologia , Vírus de Plantas/patogenicidade , Animais , Feminino , Fertilidade/fisiologia , Hemípteros/fisiologia , Oócitos/citologia , Ovário/anatomia & histologia
3.
BMB Rep ; 41(5): 388-93, 2008 May 31.
Artigo em Inglês | MEDLINE | ID: mdl-18510870

RESUMO

The Pphsc70 (heat shock cognate 70) gene was isolated from the endoparasitoid Pteromalus puparum and then characterized. The full-length cDNA was 2204 base pair (bp) and contained a single 1968 bp ORF that encoded a polypeptide of 656 amino acids with a predicted molecular mass of 71.28 kDa. Phylogenetic analysis based on Hsc70 amino acid sequences from fifteen insect species agreed with the present phylogeny. In addition, genomic DNA confirmed the presence of three introns located at the coding region as well as the 5'UTR. A significant elevation of Pphsc70 expression was observed following heat treatment, however, continued exposure to heat shock or recovery caused the expression of induced mRNA to gradually decline to levels that were significantly lower than those of control pupae (P < 0.05). In addition, a significant increase was observed in the emergence rate of pupae that were preheated at 40 degrees C and then exposed to 50 degrees C for 1 h when compared with the pupae that were not preheated, but instead directly exposed to 50 degrees C. Taken together, these results revealed that exposure to gradually increasing temperatures can enhance an insects thermo-tolerance.


Assuntos
Proteínas de Choque Térmico HSC70/genética , Proteínas de Insetos/genética , Vespas/genética , Vespas/fisiologia , Animais , Sequência de Bases , Regulação da Expressão Gênica , Proteínas de Choque Térmico HSC70/classificação , Proteínas de Choque Térmico HSC70/fisiologia , Temperatura Alta , Proteínas de Insetos/fisiologia , Modelos Genéticos , Filogenia , Reação em Cadeia da Polimerase , Vespas/metabolismo
4.
J Insect Physiol ; 53(5): 468-77, 2007 May.
Artigo em Inglês | MEDLINE | ID: mdl-17368664

RESUMO

Vitellin (Vt) and vitellogenin (Vg) profiles were analyzed in Pteromalus puparum, a pupal endoparasitoid of Pieris rapae. Non-denaturing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analyses indicated that both native Vt and Vg were likely 370 kDa in size, consisting of two subunits of approximate 206 and 165 kDa. An indirect double antibody enzyme-linked immunosorbent assay (ELISA) for monitoring hemolymph Vg and ovarian Vt levels was developed using a monoclonal antibody and a polyclonal antibody made specially against P. puparum Vt. The synthesis and uptake of Vg in this wasp was initiated immediately after adult eclosion. The hemolymph Vg and ovarian Vt reached their highest level of 0.58 and 4.51 microg per female 24 and 48 h after adult eclosion, respectively. Both Vg synthesis and uptake were in parallel with ovarian development. The Vt levels in the developing embryos decreased progressively except 12h after parasitism. Meanwhile, nine new polypeptides with sizes ranging from 59.2 to 151 kDa, possibly resulting from the limited proteolysis of Vt originally accumulated in newly laid eggs, were detected de-novo during embryonic development using Western blotting with the monoclonal antibody against Vt. These studies provide the basis for future investigation into endocrinal regulations of vitellogenesis and understanding the reproductive strategy in this wasp.


Assuntos
Vitelinas/isolamento & purificação , Vitelogênese/fisiologia , Vespas/metabolismo , Animais , Anticorpos Monoclonais/química , Borboletas/parasitologia , Embrião não Mamífero/metabolismo , Feminino , Interações Hospedeiro-Parasita/fisiologia , Pupa/parasitologia , Fatores de Tempo , Vitelinas/química , Vitelinas/metabolismo , Vespas/fisiologia
6.
Brain Res ; 980(1): 11-23, 2003 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-12865155

RESUMO

Aluminum exposure and apoptotic cell death has been implicated in several neurodegenerative diseases. The mechanisms by which aluminum interacts with the nervous system are only partly understood. In this study, we used cultured cortical neurons to investigate the ability of aluminum to induce the apoptosis of neurons and to explore the role of SAPK/JNK (stress-activated protein kinase or c-jun N-terminal kinase) signal transduction pathway on the apoptosis induced by aluminum. We found that aluminum-induced degeneration of cortical neurons involved the DNA fragmentation characteristic of apoptosis, and staining of aluminum-treated neurons with the DNA-binding fluorochrome Hoechst 33258 revealed the typical apoptotic condensation and fragmentation of chromatin. The rate of apoptosis increased significantly (from 4.9 to 13.1, 21.4, and 59.8%, P<0.01), which was measured by TdT-mediated dUTP nick end labeling. Western blot analysis showed that SAPK/JNK activities of cortical neurons varies when the exposure time of AlCl(3) were different. The phosphorylation levels were 4.2, 3.3, 1.9 and 1.1 times greater compared to control cultures for 6, 12, 24, and 48 h, respectively (P<0.01). Furthermore, a JNK pathway inhibitor, CEP-11004 (KT8138) inhibited the activation of SAPK/JNK to protect cortical neurons from apoptosis induced by aluminum chloride. Our study demonstrates that aluminum can induce the apoptosis of cortical neurons and SAPK/JNK signal transduction pathway may play an important role in the apoptosis.


Assuntos
Alumínio/efeitos adversos , Apoptose , Córtex Cerebral/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Animais , Western Blotting , Carbazóis/farmacologia , Técnicas de Cultura de Células , Córtex Cerebral/enzimologia , Fragmentação do DNA/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Indóis/farmacologia , Proteínas Quinases JNK Ativadas por Mitógeno , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Neurônios/enzimologia , Fosforilação/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley
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