Genetic transformation of sweet oranges with the D4E1 gene driven by the AtPP2 promoter

Attílio, Lísia Borges; Filho, Francisco de Assis Alves Mourão; Harakava, Ricardo; Silva, Tatiane Loureiro da; Miyata, Luzia Yuriko; Stipp, Liliane Cristina Liborio; Mendes, Beatriz Madalena Januzzi

doi:10.1590/s0100-204x2013000700006

Cited by 12 publications

(10 citation statements)

References 27 publications

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“…was\10 lM. We found that D4E1 was most effective against A. flavus in vitro, as previously reported (De Lucca et al 1998), but the transient expression of this peptide in tobacco plants induced severe necrosis, in contrast to earlier reports (Attílio et al 2013;Cary et al 2000;Mentag et al 2003;Rajasekaran et al 2005). We therefore tested the activity of thanatin against A. flavus and A. parasiticus and for the first time demonstrated its antifungal properties against both species in vitro and against A. flavus in planta.…”

Section: Discussionsupporting

confidence: 54%

Thanatin confers partial resistance against aflatoxigenic fungi in maize (Zea mays)

et al. 2015

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Aflatoxin-producing fungi can contaminate plants and plant-derived products with carcinogenic secondary metabolites that present a risk to human and animal health. In this study, we investigated the effect of antimicrobial peptides on the major aflatoxigenic fungi Aspergillus flavus and A. parasiticus. In vitro assays with different chemically-synthesized peptides demonstrated that the broad-spectrum peptide thanatin from the spined soldier bug (Podisus maculiventris) had the greatest potential to eliminate aflatoxigenic fungi. The minimal inhibitory concentrations of thanatin against A. flavus and A. parasiticus were 3.13 and 12.5 µM, respectively. A thanatin cDNA was subsequently cloned in a plant expression vector under the control of the ubiquitin-1 promoter allowing the recombinant peptide to be directed to the apoplast in transgenic maize plants. Successful integration of the thanatin expression cassette was confirmed by PCR and expression was demonstrated by semi-quantitative RT-PCR in transgenic maize kernels. Infection assays with maize kernels from T1 transgenic plants showed up to three-fold greater resistance against Aspergillus spp. infections compared to non-transgenic kernels. We demonstrated for the first time that heterologous expression of the antimicrobial peptide thanatin inhibits the growth of Aspergillus spp. in transgenic maize plants offering a solution to protect crops from aflatoxin-producing fungi and the resulting aflatoxin contamination in the field and under storage conditions.

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Section: Discussionsupporting

confidence: 54%

Thanatin confers partial resistance against aflatoxigenic fungi in maize (Zea mays)

et al. 2015

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“…The AtPP2 promoter was used to govern transcription of a gene for synthetic antimicrobial peptide D4E1 to control the HLB-associated bacteria in citrus (Attílio et al 2013 ). Among the main diseases, citrus variegated chlorosis (CVC), caused by Xylella fastidiosa , is characterized by colonization and clogging of the xylem vessels.…”

Section: Sweet Orangementioning

confidence: 99%

Promoters for Transgenic Horticultural Plants

Smirnova

Tishchenko²,

Ermakov

et al. 2014

Abiotic Stress Biology in Horticultural Plants

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Gene engineering provides an opportunity to obtain plants with either silenced or overexpressed target genes. This approach is frequently used to study various aspects of the biochemistry, physiology, and stress tolerance of horticultural plants. Selection of appropriate promoters to govern transcription of a transgenic construct is an important step in the development of effi cient genetic models. Here we present a brief overview of available literature data on the promoters investigated with the transgenic horticultural plants and some valuable information resources in this fi led.

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“…However, strong transgene expression without tissue discrimination can result in unnecessary energy expenditure by the plant and possible side effects due to putatively harmful transgene products (ALTPETER et al, 2005;WANG et al, 2005;ZHAO et al, 2004). One method to fine tune transgene expression is to use tissue-specific promoters induced by pathogens or abiotic stresses (BARBOSA-MENDES et al, 2009;DUTT et al, 2012;MIYATA et al, 2012;BENYON et al, 2013;ATTÍLIO et al, 2013;ZOU et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

PHLOEM PROMOTERS IN TRANSGENIC SWEET ORANGE ARE DIFFERENTIALLY TRIGGERED BY Candidatus Liberibacter asiaticus

et al. 2017

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-The use of promoters preferentially expressed in specific plant tissues is a desirable strategy to search for resistance for pathogens that colonize these tissues. The bacterium Candidatus Liberibacter asiaticus (Las), associated with huanglongbing disease (HLB) of citrus, colonizes phloem vessels. Some promoters, besides conferring tissue-specific expression, can also respond to the presence of the pathogen. The objective of the present study was to verify if the presence of Las could modulate the activation of the phloem-specific promoters AtPP2 (Arabidopsis thaliana phloem protein 2), AtSUC2 (A. thaliana sucrose transporter 2) and CsPP2 (Citrus phloem protein 2), known to be expressed in Citrus sinensis phloem. 'Hamlin' sweet orange plants (Citrus sinensis L. Osbeck) transformed with the uidA (GUS) reporter gene under the control of AtPP2, AtSUC2 and CsPP2 promoters were infected to evaluate the interdependence between transgene expression and the concentration of Las. Plants were inoculated with Las by Diaphorina citri and eighteen months later, bacterial concentration and uidA expression were determined by qPCR and RT-qPCR, respectively. Reporter gene expression driven by AtSUC2 promoter was strongly and positively correlated with Las concentration. Therefore, this promoter combines desirable features of both tissue-specificity and pathogen-inducibility for the production of transgenic plants tolerant to Las. Index terms: Citrus sinensis, Diaphorina citri, genetic transformation, GUS, huanglongbing, quantitative real-time PCR. PROMOTORES DE FLOEMA SÃO DIFERENTEMENTE ATIVADOS POR Candidatus Liberibacter asiaticus EM LARANJEIRAS DOCES TRANSGÊNICASRESUMO -A utilização de promotores preferencialmente expressos em tecidos vegetais específicos é uma estratégia desejável na busca por resistência a patógenos que colonizam tais tecidos. A bacteria Candidatus Liberibacter asiaticus (Las), associada à doença huanglongbing (HLB) em citros, coloniza o tecido floemático. Alguns promotores, além de conferir expressão específica em certos tecidos, podem também responder diferencialmente à presença de patógenos. O objetivo deste estudo foi verificar se a presença de Las poderia modular a ativação dos promotores de floema AtPP2 (Arabidopsis thaliana phloem protein 2), AtSUC2 (A. thaliana sucrose transporter 2) e CsPP2 (Citrus phloem protein 2), conhecidos por expressão específica em floema de Citrus sinensis. Plantas de laranja 'Hamlin' (Citrus sinensis L. Osbeck) transformadas com o gene reporter uidA (GUS) sob controle dos promotores AtPP2, AtSUC2 e CsPP2 foram infectadas para avaliar a interdependência entre a expressão dos transgenes e concentração de Las. Plantas foram inoculadas com Las por meio de Diaphorina citri e dezoito meses após, a concentração de bactéria e expressão do gene uidA foram determinadas por qPCR e RT-qPCR, respectivamente. A expressão do gene reporter dirigida pelo promotor AtSUC2 foi forte e positivamente correlacionada com a concentração de Las. Portanto, este promotor contém característ...

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Genetic transformation of sweet oranges with the D4E1 gene driven by the AtPP2 promoter

Cited by 12 publications

References 27 publications

Thanatin confers partial resistance against aflatoxigenic fungi in maize (Zea mays)

Thanatin confers partial resistance against aflatoxigenic fungi in maize (Zea mays)

Promoters for Transgenic Horticultural Plants

PHLOEM PROMOTERS IN TRANSGENIC SWEET ORANGE ARE DIFFERENTIALLY TRIGGERED BY Candidatus Liberibacter asiaticus

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