The ecdysteroid UDP-glucosyltransferase gene of Autographa californica nucleopolyhedrovirus alters the moulting and metamorphosis of a non-target insect, the silkworm, Bombyx mori (Lepidoptera, Bombycidae).

Shikata, Masuzo; Shibata, Heki; Sakurai, Makiko; Sano, Yoshitaka; Hashimoto, Yoshifumi; Matsumoto, Tsuguo

doi:10.1099/0022-1317-79-6-1547

Cited by 29 publications

(21 citation statements)

References 20 publications

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“…The average durations of the larval and pupal stages in the uninfected animals were 10 and 15 days, respectively, but the duration of the pupal stage of infected animals was a few days longer than in the uninfected animals. This phenomenon suggested that the ecdysteroid UDP-glucosyltransferase (EGT) gene of AcNPV was expressed, and the secreted EGT altered growth of the infected animals (Shikata et al 1998). After infection with the recombinant baculovirus, no progeny virus was detected in hemolymph at 5, 10, or 15 days postinoculation (p.i.…”

Section: Resultsmentioning

confidence: 99%

Gene targeting in the silkworm by use of a baculovirus

Yamao¹,

Katayama²,

Nakazawa³

et al. 1999

Genes & Development

104

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The Bombyx mori fibroin light (L)-chain gene was cloned and the green fluorescent protein (GFP) gene inserted into exon 7. The chimeric L-chain-GFP gene was used to replace the polyhedrin gene of Autographa californica nucleopolyhedrovirus (AcNPV). This recombinant virus was used to target the L-chain-GFP gene to the L-chain region of the silkworm genome. Female moths were infected with the recombinant virus and then mated with normal male moths. Genomic DNA from their progenies was screened for the desired targeting event. This analysis showed that the chimeric gene had integrated into the L-chain gene on the genome by homologous recombination and was stably transmitted through generations. The chimeric gene was expressed in the posterior silk gland, and the gene product was spun into the cocoon layer.

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Section: Resultsmentioning

confidence: 99%

Gene targeting in the silkworm by use of a baculovirus

Yamao¹,

Katayama²,

Nakazawa³

et al. 1999

Genes & Development

104

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“…O'Reilly (1995) suggested that the baculovirus EGT increased progeny virus production by prolonging the feeding period of the host insect and viral propagation efficiency by altering its behavioral patterns to remain at host plants until its death. Because effects of N. rileyi conditioned medium injection into silkworm were very similar to those of AcNPV infection (Shikata et al, 1998), ecdysteroid 22-oxidase of N. rileyi may have the same biological functions. More importantly, inhibition of molting appears to play a significant role in establishing successful infection of this fungus.…”

Section: Archives Of Insect Biochemistry and Physiologymentioning

confidence: 98%

Entomogenous fungus Nomuraea rileyi inhibits host insect molting by C22‐oxidizing inactivation of hemolymph ecdysteroids

Kiuchi

Yasui

Hayasaka

et al. 2002

Arch Insect Biochem Physiol

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The entomogenous fungus Nomuraea rileyi reportedly secretes a proteinaceous substance inhibiting larval molt and metamorphosis in the silkworm Bombyx mori. We studied the possibility that N. rileyi controls B. mori development by inactivating hemolymph molting hormone, ecdysteroids. Incubation of ecdysone (E) and 20-hydroxyecdysone (20E) in fungal-conditioned medium resulted in their rapid modification into products with longer retention times in reverse-phase HPLC. Each modified product from E and 20E was purified by HPLC, and identified by NMR as 22-dehydroecdysone and 22-dehydro-20-hydroxyecdysone. Some other ecdysteroids with a hydroxyl group at position C22 were also modified. Injection of the fungal-conditioned medium into Bombyx mori larvae in the mid-4th instar inhibited larval molt but induced precocious pupal metamorphosis, and its injection into 5th instar larvae just after gut purge blocked pupal metamorphosis. In hemolymph of injected larvae, E and 20E disappeared and, in turn, 22-dehydroecdysone and 22-dehydro-20-hydroxyecdysone accumulated. These results indicate that N. rileyi secretes a specific enzyme that oxidizes the hydroxyl group at position C22 of hemolymph ecdysteroids and prevents molting in B. mori larvae. Abbreviations used: E = ecdysone; EGT = ecdysteroid UDP-glucosyltransferase; ME = modified ecdysone; M20E = modified 20-hydroxyecdysone; NPV = nucleopolyhedrovirus; 20E = 20-hydroxyecdysone.

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“…It has been reported previously that the proliferation of B. mori AcNPV is related to the virus concentration (Shikata et al, 1998;Yamao et al, 1999). To investigate this, we compared the proliferation of B. mori AcNPV in both HPS and LPS, both in terms of the duration since injection, and the initial concentration of virus used.…”

Section: Discussionmentioning

confidence: 99%

“…So, the baculovirus expression system, AcNPV, is commonly applied for large scale expression for eukaryotic proteins in permissive cell lines or insects (Maeda, 1989). In AcNPV hosted by A. californica has been used to infect cultured cells derived from B. mori, but they can fail to breed (Shikata et al, 1998;Yamao et al, 1999). Recently however, the existence of a B. mori strain that enabled the breeding of AcNPV has been reported Lee et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Screening of silkworm strains for efficient recombinant protein production by Autographa californica nucleopolyhedrosis virus (AcNPV)

Park¹,

Kim²,

Kang³

et al. 2014

International Journal of Industrial Entomology

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Baculoviruses base vectors come to be regarded as methods for in vivo gene delivery and transient expression to the silkworm. In the case of silkworm, B. mori, two types of baculoviruses, AcNPV (Autographa californica nuclear polyhedrosis virus) and BmNPV (Bombyx mori nuclear polyhedrosis virus), are potentially applicable as vectors. Recently, AcNPV showed promising results with some silkworm strains despite different hostspecificities. We searched for a highly-permissive silkworm strain in the B. mori stocks of Kyungpook National University that could produce high levels of recombinant protein. Seventy strains were screened using the recombinant AcNPV/BmA3-Luc virus. Based on the measured luciferase activity, the strains could be divided into three groups, high-, middle-, and low-permissive strains, according to their relative recombinant protein expression levels. At 48 hours post-injection, the luciferase activity in the high-permissive strains was 500-fold greater than that of the low-permissive strains. At 72 hours post-injection, a significant elevation in luciferase activity was observed in the hemocytes of all strains. Then, based on the above results, the High Permissive Strain (HPS) S10 and the Low Permissive Strain (LPS) S39 were pick up and was carried out Dot blotting, RT-PCR and Real time PCR.

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The ecdysteroid UDP-glucosyltransferase gene of Autographa californica nucleopolyhedrovirus alters the moulting and metamorphosis of a non-target insect, the silkworm, Bombyx mori (Lepidoptera, Bombycidae).

Cited by 29 publications

References 20 publications

Gene targeting in the silkworm by use of a baculovirus

Gene targeting in the silkworm by use of a baculovirus

Entomogenous fungus Nomuraea rileyi inhibits host insect molting by C22‐oxidizing inactivation of hemolymph ecdysteroids

Screening of silkworm strains for efficient recombinant protein production by Autographa californica nucleopolyhedrosis virus (AcNPV)

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