Characterization of cDNAs encoding three trypsin-like proteinases and mRNA quantitative analysis in Bt-resistant and -susceptible strains of Ostrinia nubilalis

Li, Huarong; Oppert, Brenda; Higgins, R. A.; Huang, Fangneng; Buschman, Lawrent L.; Gao, Jianrong; Zhu, Kun Yan

doi:10.1016/j.ibmb.2005.03.004

Cited by 50 publications

(78 citation statements)

References 63 publications

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“…The midgut proteinase of Lepidoptera larvae is widely studied because of its important role in digestion (Li et al, 2005). Three days after ingesting tissue of whorl leaves or ears of two transgenic maize lines, the in vivo activity of carboxylesterase in the first three larval instars of ACB was significantly decreased, which is consistent with the results obtained by Xu et al (2006).…”

Section: Discussionsupporting

confidence: 89%

Effect of insect-resistant transgenic maize on growth and development, utilization of nutrients and in vivo activity of the detoxification enzymes of the Asian corn borer, Ostrinia furnacalis (Lepidoptera: Pyralidae)

Shi¹,

Lu²,

Shi³

et al. 2011

Eur. J. Entomol.

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Abstract. There is little systematic research on the effect of transgenic maize expressing Bacillus thuringiensis (Bt) toxins on insect survival and the response in vivo of their detoxification enzymes. Results of laboratory bioassays indicate that the whorl leaves of transgenic maize lines G03-2396 and G03-2739 significantly affected the survival of neonate and third instar larvae of the Asian corn borer, Ostrinia furnacalis (Güenée). Neonate mortality two days after being fed on G03-2739 was 72.5% and exceeded 92.5% four days later. The percentage mortality of third instar larvae six days after being fed whorl leaves from G03-2396 exceeded 65%. The resistance of whorl leaves to insect attack was greater than that of maize-ears, but less effective against attacks by third instar larvae. Neonate development was more prolonged when they were fed on whorl leaves of both the transgenic maize lines. In contrast, low pupation (<80% of that recorded in the controls), ecolosion and fecundity were recorded following ingestion of maizeears, with pupal weights 10-14 mg lower than that of controls. The growth rates of third instar larvae recorded three and six days after feeding on whorl leaves were lower, but food utilization, conversion and relative metabolism were not significantly affected. Relative food ingestion, relative growth rate and other nutritional indicators of third instar larvae were significantly lower six days after the ingestion of ears and that of fifth instar larvae after three days. In vivo activity of carboxylesterase was decreased to a greater degree after three days when the larvae were fed on whorl leaves than ears, with no significant effect on fifth instar larvae. Furthermore, in vivo activity of glutathione S-transferase (GST) of third instar larvae was significantly affected following the ingestion of whorl leaves. These results are discussed in the context of the literature on the resistance of transgenic plants and of improving the resistance of plants to attack by the different larval stages of insects. 547* Corresponding author; e-mail: yzyang@yzu.edu.cn insect interactions and their potential application in the development of new Bt crops for use in the control of insect pests. MATERIAL AND METHODSABBREVIATIONS: ACB -Asian corn borer; BSA -bovine serum albumin; CDNB -2, 4-dinitrochlorobenzene; GSH -glutathione; GST -glutathione S-transferase; L1 -ACB neonates; L3 -third instar ACB larvae; L5 -fifth instar ACB larvae; OD600 -optical density at 600 nm. Test maize lines and planting methodsTwo maize lines, G03-2396 and G03-2739, which contain Cry1Ab insecticidal genes, were provided by the China Agricultural University (Beijing, China). The conventional maize cultivar, Suyu 16, served as the control, and was provided by the Maize Breeding Group of Yangzhou University (Yangzhou, China). The three maize lines were all planted in an experimental plot in the agricultural pastures of Yangzhou University. Following the normal method of planting in the Yangtze River Valley of China, the maize was p...

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Section: Discussionsupporting

confidence: 89%

Effect of insect-resistant transgenic maize on growth and development, utilization of nutrients and in vivo activity of the detoxification enzymes of the Asian corn borer, Ostrinia furnacalis (Lepidoptera: Pyralidae)

Shi¹,

Lu²,

Shi³

et al. 2011

Eur. J. Entomol.

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“…This strain showed a broad spectrum of resistance to Bt protoxins, being highly resistant to all Bt protoxins present in Dipel (Cry1Aa, Cry1Ab, Cry1Ac, and Cry2Aa), cross-resistant to Cry1Ba protoxin (27), and only marginally resistant to activated Cry1Ab (26). Resistance was associated with reduction of a trypsin-like protease involved in activation of the Bt protoxin form (26,29), and no alteration in Cry1Ab and Cry1Ac binding was found in the resistant insects (28). Because the Dipel formulation contains multiple toxins, it is unclear whether such a resistance mechanism would evolve in response to selection from a single toxin.…”

Section: Discussionmentioning

confidence: 99%

“…kurstaki in O. nubilalis has been reported in laboratory-selected strains exposed to the Bt formulation Dipel ES (18) and to the isolated Cry1Ac (3) and Cry1Ab (5, 39) toxins. A resistance mechanism has been partially characterized only in the Dipel-selected strain, in which differences in midgut proteases were linked to resistance (26,29).…”

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confidence: 99%

Analyses of Cry1Ab Binding in Resistant and Susceptible Strains of the European Corn Borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae)

Siqueira

González‐Cabrera

Ferré

et al. 2006

Appl Environ Microbiol

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Cry1Ab toxin binding analysis was performed to determine whether resistance in laboratory-selected Ostrinia nubilalis strains is associated with target site alteration. Brush border membrane vesicles were prepared using dissected midguts from late instars of susceptible and resistant strains (Europe-R and RSTT) of O. nubilalis. Immunoblot analysis indicated that three different proteins bound to Cry1Ab toxin and were recognized by an anticadherin serum. In a comparison of resistant and susceptible strains, reduced Cry1Ab binding was apparent for all three bands corresponding to cadherin-like proteins in the Europe-R strain, while reduced binding was apparent in only one band for the RSTT strain. Real-time analysis of Cry1Ab binding to gut receptors using surface plasmon resonance suggested slight differences in affinity in both resistant strains. Additional binding analysis was conducted using 125 I-labeled Cry1Ab, Cry1Ac, and Cry1Aa. Slight differences were again observed between the resistant and susceptible strains for Cry1Ab binding. However, when binding of 125 I-labeled Cry1Aa was tested, a 10-fold reduction in the concentration of binding sites was observed in the Europe-R strain. Expression of the O. nubilalis cadherin gene was similar in both the resistant and susceptible strains and did not account for differences in binding. In combination, the results of the present work suggest that differences in susceptibility to Cry1A toxins in the Europe-R strain of O. nubilalis are associated with altered receptor binding, although the precise nature of this mechanism is still uncertain.

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“…Fluorescence intensity was measured after each round of PCR. Mean threshold PCR cycles of our samples were calculated from the three independent trials and normalized to the quantities of 18S rRNA (RPS 3; Li et al 2005). Starting quantities for each sample were calculated by using the linear standard equation formulated from starting quantities and the mean log threshold ßuorescence values obtained from standards.…”

Section: Figmentioning

confidence: 99%

On-Plant Selection and Genetic Analysis of European Corn Borer (Lepidoptera: Crambidae) Behavioral Traits: Plant Abandonment Versus Plant Establishment

et al. 2014

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Although some studies have investigated how insect behavior could influence resistance evolution to transgenic plants, none have determined if behavioral traits respond to selection pressure and how they may be inherited. We investigated plant establishment and abandonment traits for the European corn borer, Ostrinia nubilalisi (Hübner) (Lepidoptera: Crambidae), by conducting a laboratory selection experiment and quantifying patterns of gene expression. Egg masses with emerging larvae were placed on maize plants and silking individuals were collected every 15 min during a 4-h period to generate a plant abandonment (PA) colony. Plants were dissected 24-72 h later, and larvae were collected for a plant establishment colony. Selection of the PA colony showed an increased propensity to abandon the host plant by the third generation. The propensity for larvae to establish on the plants, however, did not show a significant response until the sixth generation. Quantitative real-time-polymerase chain reaction (qRT-PCR) was used to determine expression profiles for behavior associated genes (foraging and Onslmo). Egg samples from the two selected colonies and nonselected laboratory colony were collected at 0, 24, 48, 72, and 96 h after egg deposition, and first instars were sampled after exposure to maize tissue. Compared with the plant establishment and nonselected laboratory colonies at the 0-h time period, foraging and Onslmoshowed higher expression in the PA colony. This is the first study that has specifically selected for these traits over several generations and analyzed behavior-associated genes to elucidate genetic changes. RightsWorks produced by employees of the U.S. Government as part of their officials duties are not copyrighted within the U.S. The content of this document is not copyrighted. ABSTRACT Although some studies have investigated how insect behavior could inßuence resistance evolution to transgenic plants, none have determined if behavioral traits respond to selection pressure and how they may be inherited. We investigated plant establishment and abandonment traits for the European corn borer, Ostrinia nubilalisi (Hü bner) (Lepidoptera: Crambidae), by conducting a laboratory selection experiment and quantifying patterns of gene expression. Egg masses with emerging larvae were placed on maize plants and silking individuals were collected every 15 min during a 4-h period to generate a plant abandonment (PA) colony. Plants were dissected 24 Ð72 h later, and larvae were collected for a plant establishment colony. Selection of the PA colony showed an increased propensity to abandon the host plant by the third generation. The propensity for larvae to establish on the plants, however, did not show a signiÞcant response until the sixth generation. Quantitative real-timeÐpolymerase chain reaction (qRT-PCR) was used to determine expression proÞles for behavior associated genes (foraging and Onslmo). Egg samples from the two selected colonies and nonselected laboratory colony were collected at 0, 24...

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Characterization of cDNAs encoding three trypsin-like proteinases and mRNA quantitative analysis in Bt-resistant and -susceptible strains of Ostrinia nubilalis

Cited by 50 publications

References 63 publications

Effect of insect-resistant transgenic maize on growth and development, utilization of nutrients and in vivo activity of the detoxification enzymes of the Asian corn borer, Ostrinia furnacalis (Lepidoptera: Pyralidae)

Effect of insect-resistant transgenic maize on growth and development, utilization of nutrients and in vivo activity of the detoxification enzymes of the Asian corn borer, Ostrinia furnacalis (Lepidoptera: Pyralidae)

Analyses of Cry1Ab Binding in Resistant and Susceptible Strains of the European Corn Borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae)

On-Plant Selection and Genetic Analysis of European Corn Borer (Lepidoptera: Crambidae) Behavioral Traits: Plant Abandonment Versus Plant Establishment

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