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Molecular cloning and RNA interference-mediated functional characterization of a Halloween gene spook in the white-backed planthopper Sogatella furcifera.

ABSTRACT: BACKGROUND: Ecdysteroid hormones ecdysone and 20-hydroxyecdysone play fundamental roles in insect postembryonic development and reproduction. Five cytochrome P450 monooxygenases (CYPs), encoded by Halloween genes, have been documented to be involved in the ecdysteroidogenesis in insect species of diverse orders such as Diptera, Lepidoptera and Orthoptera. Up to now, however, the involvement of the Halloween genes in ecdysteroid synthesis has not been confirmed in hemipteran insect species. RESULTS: In the present paper, a Halloween gene spook (Sfspo, Sfcyp307a1) was cloned in the hemipteran Sogatella furcifera. SfSPO has three insect conserved P450 motifs, i.e., Helix-K, PERF and heme-binding motifs. Temporal and spatial expression patterns of Sfspo were evaluated by qPCR. Sfspo showed three expression peaks in late second-, third- and fourth-instar stages. In contrast, the expression levels were lower and formed three troughs in the newly-molted second-, third- and fourth-instar nymphs. On day 3 of the fourth-instar nymphs, Sfspo clearly had a high transcript level in the thorax where PGs were located. Dietary introduction of double-stranded RNA (dsRNA) of Sfspo into the second instars successfully knocked down the target gene, and greatly reduced expression level of ecdysone receptor (EcR) gene. Moreover, knockdown of Sfspo caused lethality and delayed development during nymphal stages. Furthermore, application of 20-hydroxyecdysone on Sfspo-dsRNA-exposed nymphs did not increase Sfspo expression, but could almost completely rescue SfEcR expression, and relieved the negative effects on nymphal survival and development. CONCLUSION: In S. furcifera, Sfspo was cloned and the conservation of SfSPO is valid. Thus, SfSPO is probably also involved in ecdysteroidogenesis for hemiptera.

SUBMITTER: Jia S

PROVIDER: S-EPMC3766648 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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Molecular cloning and RNA interference-mediated functional characterization of a Halloween gene spook in the white-backed planthopper Sogatella furcifera.

Jia Shuang S Wan Pin-Jun PJ Zhou Li-Tao LT Mu Li-Li LL Li Guo-Qing GQ

BMC molecular biology 20130904

<h4>Background</h4>Ecdysteroid hormones ecdysone and 20-hydroxyecdysone play fundamental roles in insect postembryonic development and reproduction. Five cytochrome P450 monooxygenases (CYPs), encoded by Halloween genes, have been documented to be involved in the ecdysteroidogenesis in insect species of diverse orders such as Diptera, Lepidoptera and Orthoptera. Up to now, however, the involvement of the Halloween genes in ecdysteroid synthesis has not been confirmed in hemipteran insect species ...[more]

PMID: 24007644

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Project description:Rice (Oryza sativa L.) is one of the essential staple foods for more than half of the world's population, and its production is affected by different environmental abiotic and biotic stress conditions. The white-backed planthopper (WBPH, Sogatella furcifera) causes significant damage to rice plants, leading to substantial economic losses due to reduced production. In this experiment, we applied exogenous hormones (gibberellic acid and methyl jasmonate) to WBPH-infested rice plants and examined the relative expression of related genes, antioxidant accumulation, the recovery rate of affected plants, endogenous hormones, the accumulation of H2O2, and the rate of cell death using DAB and trypan staining, respectively. The expression of the transcriptional regulator (OsGAI) and gibberellic-acid-mediated signaling regulator (OsGID2) was upregulated significantly in GA 50 µM + WBPH after 36 h. OsGAI was upregulated in the control, GA 50 µM + WBPH, GA 100 µM + WBPH, and MeJA 100 µM + WBPH. However, after 48 h, the OsGID2 was significantly highly expressed in all groups of plants. The glutathione (GSH) values were significantly enhanced by GA 100 µM and MeJA 50 µM treatment. Unlike glutathione (GSH), the catalase (CAT) and peroxidase (POD) values were significantly reduced in control + WBPH plants. However, a slight increase in CAT and POD values was observed in GA 50 + WBPH plants and a reduction in the POD value was observed in GA 100 µM + WBPH and MeJA 50 µM + WBPH plants. GA highly recovered the WBPH-affected rice plants, while no recovery was seen in MeJA-treated plants. MeJA was highly accumulated in control + WBPH, MeJA 50 µM + WBPH, and GA 100 µM + WBPH plants. The H2O2 accumulation was highly decreased in GA-treated plants, while extensive cell death was observed in MeJA-treated plants compared with GA-treated plants. From this study, we can conclude that the exogenous application of GA can overcome the effects of the WBPH and enhance resistance in rice.

Dataset Information

Molecular cloning and RNA interference-mediated functional characterization of a Halloween gene spook in the white-backed planthopper Sogatella furcifera.

Publications

Molecular cloning and RNA interference-mediated functional characterization of a Halloween gene spook in the white-backed planthopper Sogatella furcifera.

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