Beta vulgaris L. serine proteinase inhibitor gene expression in insect resistant sugar beet

Savić﻿﻿﻿, Jelena; Smigocki, Ann C.

doi:10.1007/s10681-012-0679-z

Cited by 7 publications

(13 citation statements)

References 45 publications

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“…In addition to the 30 kDa BvSTI protein activity previously observed in sugar beet, two additional zones of activity corresponding to approximately 28 and 26 kDa were clearly visible in line 11-4, 11-5 and 11-13 (Fig. 3B, lane 1, 2 and 4) [49]. Transformant 11-6 had reduced level of the active 26 kDa trypsin inhibitor (Fig.…”

Section: Resultsmentioning

confidence: 52%

“…3A) [49]. Weak protein signal of about 30 kDa as well as a stronger signal in the range of 22–25 kDa cross-reacted with the BvSTI antibodies in the transgenic 11-4, 11-6, 11-13 and 12-2 tobacco plants (Fig.…”

Section: Resultsmentioning

confidence: 88%

“…In a study of sugar beet root defense responses, a single serine PI gene ( BvSTI ) was identified among the more than 150 sugar beet genes whose expression was found to be modulated by a dipteran pest of sugar beet, the root maggot [35]. Expression of the BvSTI gene was determined to be up-regulated by mechanical- and insect-wounding in sugar beet lines used in breeding for root maggot resistance [41], [49]. The observed lack or reduced accumulation and activity of BvSTI PI in tissues of susceptible and less resistant lines emphasized the potentially important role of the BvSTI PI in insect pest defense mechanisms.…”

Section: Discussionmentioning

confidence: 99%

“…Following transfer, membranes were rinsed in deionized water and gently agitated in blocking solution (5% BLOT-QuickBlocker, Chemicon International) for 1 hour. Membranes were then incubated with rabbit anti-BvSTI antibodies produced to a mixture of two most antigenic BvSTI peptides spanning amino acids 28–41 and 69–82 (GenScript Corporation, NJ, USA) [49] at 1∶2000 or 1∶5000 (v/v) dilutions in 1 x TBS-T (0.137 M NaCl, 0.02 M Tris pH 7.6, 0.1% Tween 20). After 1 hour 30 minutes incubation, membranes were rinsed two times in 1 × TBS-T for 10 minutes each, and incubated for 1 hour in alkaline phosphatase conjugated secondary antibody (AP Conjugated Goat anti-Rabbit IgG, 1∶5000 diluted in 1 × TBS-T, Chemicon International).…”

Section: Methodsmentioning

confidence: 99%

“…Membranes were washed in 1 × TBS-T two times for 15 minutes and then 1 minute in 1 × TBS to remove the Tween 20. Alkaline phosphatase was detected using BCIP/NBT (5-bromo-4-chloro-3-indolylphosphate p-toluidine salt and nitro-blue tetrazolium chloride, respectively, Roche) in 0.1 M Tris–HCl pH 9.5, 0.1 M NaCl, and 0.05 M MgCl 2 [42], [49]. Experiments were repeated two times.…”

Section: Methodsmentioning

confidence: 99%

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Pest Protection Conferred by a Beta vulgaris Serine Proteinase Inhibitor Gene

Smigocki

Ivic-Haymes

et al. 2013

PLoS ONE

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Proteinase inhibitors provide a means of engineering plant resistance to insect pests. A Beta vulgaris serine proteinase inhibitor gene (BvSTI) was fused to the constitutive CaMV35S promoter for over-expression in Nicotiana benthamiana plants to study its effect on lepidopteran insect pests. Independently derived BvSTI transgenic tobacco T2 homozygous progeny were shown to have relatively high BvSTI gene transcript levels. BvSTI-specific polyclonal antibodies cross-reacted with the expected 30 kDA recombinant BvSTI protein on Western blots. In gel trypsin inhibitor activity assays revealed a major clear zone that corresponded to the BvSTI proteinase inhibitor that was not detected in the untransformed control plants. BvSTI-transgenic plants were bioassayed for resistance to five lepidopteran insect pests. Spodoptera frugiperda, S. exigua and Manduca sexta larvae fed BvSTI leaves had significant reductions in larval weights as compared to larvae fed on untransformed leaves. In contrast, larval weights increased relative to the controls when Heliothis virescens and Agrotis ipsilon larvae were fed on BvSTI leaves. As the larvae entered the pupal stage, pupal sizes reflected the overall larval weights. Some developmental abnormalities of the pupae and emerging moths were noted. These findings suggest that the sugar beet BvSTI gene may prove useful for effective control of several different lepidopteran insect pests in genetically modified tobacco and other plants. The sugar beet serine proteinase inhibitor may be more effective for insect control because sugar beet is cropped in restricted geographical areas thus limiting the exposure of the insects to sugar beet proteinase inhibitors and build up of non-sensitive midgut proteases.

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Section: Resultsmentioning

confidence: 52%

Section: Resultsmentioning

confidence: 88%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Pest Protection Conferred by a Beta vulgaris Serine Proteinase Inhibitor Gene

Smigocki

Ivic-Haymes

et al. 2013

PLoS ONE

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Beneficial implications of sugar beet proteinase inhibitor BvSTI on plant architecture and salt stress tolerance in Lotus corniculatus L.

Savić

Nikolić

Banjac

et al. 2019

Journal of Plant Physiology

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Screening for Resistance Against the Sugarbeet Root Maggot, Tetanops myopaeformis (Diptera: Ulidiidae), Using a Greenhouse Bioassay

Daley

Wenninger

2018

Journal of Insect Science

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The sugarbeet root maggot, Tetanops myopaeformis (von Röder) (Diptera: Ulidiidae), is a major pest of sugar beet Beta vulgaris L. (Carophyllales: Amaranthaceae)in the United States and Canada. Larval feeding on roots can reduce both stand and yield. Current management practices are heavily reliant on chemical control. However, the carbamate and organophosphate insecticides that are commonly used against T. myopaeformis are being phased out of use. Host plant resistance against this pest shows promise, but difficulties with maintaining T. myopaeformis in culture have largely limited such studies to the field. A primary objective of this study was to develop protocols for rearing a laboratory colony of T. myopaeformis that would expedite assays aimed at screening for host plant resistance. Third (final) instar larvae were collected from the field and reared to the adult stage. These laboratory-reared adults laid eggs and ultimately produced a second generation of third-instar larvae in the lab. Adult flies reared from field-collected larvae were used to examine the modality of resistance of a known resistant variety by performing no-choice and paired-choice experiments alongside a susceptible variety in the greenhouse. Paired-choice tests showed no difference in oviposition rates between the two varieties, whereas no-choice tests showed significantly greater feeding damage and abundance of larvae on the susceptible variety. For the resistant variety examined here, we observed evidence of antibiosis, not antixenosis, as the putative modality of resistance. Our laboratory and greenhouse protocols can be used to expedite the process of developing varieties with resistance to this key pest of sugar beet.

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Beta vulgaris L. serine proteinase inhibitor gene expression in insect resistant sugar beet

Cited by 7 publications

References 45 publications

Pest Protection Conferred by a Beta vulgaris Serine Proteinase Inhibitor Gene

Pest Protection Conferred by a Beta vulgaris Serine Proteinase Inhibitor Gene

Beneficial implications of sugar beet proteinase inhibitor BvSTI on plant architecture and salt stress tolerance in Lotus corniculatus L.

Screening for Resistance Against the Sugarbeet Root Maggot, Tetanops myopaeformis (Diptera: Ulidiidae), Using a Greenhouse Bioassay

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