Factors influencing transient gene expression in bean (Phaseolus vulgaris L.) using an electrical particle acceleration device

Aragão, F. J. L.; de, Maria Fatima Grossi; Davey, M. R.; Brasileiro, Ana Cristina Miranda; Faria, J. C. de; Rech, E. L.

doi:10.1007/bf00236092

Cited by 39 publications

(6 citation statements)

References 17 publications

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“…Transient gene expression is influenced by several factors including species and physiological status of the explant [52], transformation parameters [53], [54], timing of gene integration [55], cell death and loss of DNA [56], [57], and gene structure [58]. PTGS also plays an important role in the post-introduction gene expression decline, so-called transient expression [39], [59].…”

Section: Discussionmentioning

confidence: 99%

Enhanced Transgene Expression in Sugarcane by Co-Expression of Virus-Encoded RNA Silencing Suppressors

Gao

Damaj²,

Park³

et al. 2013

PLoS ONE

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Post-transcriptional gene silencing is commonly observed in polyploid species and often poses a major limitation to plant improvement via biotechnology. Five plant viral suppressors of RNA silencing were evaluated for their ability to counteract gene silencing and enhance the expression of the Enhanced Yellow Fluorescent Protein (EYFP) or the β-glucuronidase (GUS) reporter gene in sugarcane, a major sugar and biomass producing polyploid. Functionality of these suppressors was first verified in Nicotiana benthamiana and onion epidermal cells, and later tested by transient expression in sugarcane young leaf segments and protoplasts. In young leaf segments co-expressing a suppressor, EYFP reached its maximum expression at 48–96 h post-DNA introduction and maintained its peak expression for a longer time compared with that in the absence of a suppressor. Among the five suppressors, Tomato bushy stunt virus-encoded P19 and Barley stripe mosaic virus-encoded γb were the most efficient. Co-expression with P19 and γb enhanced EYFP expression 4.6-fold and 3.6-fold in young leaf segments, and GUS activity 2.3-fold and 2.4-fold in protoplasts compared with those in the absence of a suppressor, respectively. In transgenic sugarcane, co-expression of GUS and P19 suppressor showed the highest accumulation of GUS levels with an average of 2.7-fold more than when GUS was expressed alone, with no detrimental phenotypic effects. The two established transient expression assays, based on young leaf segments and protoplasts, and confirmed by stable transgene expression, offer a rapid versatile system to verify the efficiency of RNA silencing suppressors that proved to be valuable in enhancing and stabilizing transgene expression in sugarcane.

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Section: Discussionmentioning

confidence: 99%

Enhanced Transgene Expression in Sugarcane by Co-Expression of Virus-Encoded RNA Silencing Suppressors

Gao

Damaj²,

Park³

et al. 2013

PLoS ONE

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“…Nonwrinkled seeds were then rinsed three times and soaked overnight in sterile deionized water. Apical meristems were exposed by removing cotyledons and primary leaves as described by Aragão et al (1993). Subsequently, the root tips were also excised.…”

Section: Methodsmentioning

confidence: 99%

Susceptibility of Common and Tepary Beans to Agrobacterium spp. Strains and Improvement of Agrobacterium-mediated Transformation Using Microprojectile Bombardment

Brasileiro¹,

Aragão²,

Rossi³

et al. 1996

jashs

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To develop an efficient protocol for Agrobacterium-mediated transformation of common bean (Phaseolus vulgaris L.) and tepary bean (P. acutifolius A. Gray), we have tested the susceptibility of six genotypes to eight Agrobacterium tumefaciens and two A. rhizogenes strains. The virulence of the Agrobacterium strains was shown to be genotype dependent. In general, the tumors observed on common bean cultivars were larger than those observed on tepary bean cultivars. The A. tumefaciens AT8196 and Ach5 strains and the A. rhizogenes 8196 strain induced the best responses in all genotypes tested. Polymerase chain reaction (PCR) analysis confirmed the presence of T-DNA in tumors derived from inoculation with three A. tumefaciens strains in common beans. Apical meristems of P. vulgaris cv. Jalo were bombarded with tungsten microprojectiles and then inoculated with an A. tumefaciens wild-type strain (Ach5). One month later, the explants showed a high frequency of tumor formation (50% to 70%). Similarly, when bombarded meristems were inoculated with an A. tumefaciens disarmed strain (LBA4404/p35SGUSINT), 44% of them showed substantial sectors of GUS activity, suggesting the expression of introduced gene. The bombardment/Agrobacterium system appears to be a promising method to stably transform bean through the regeneration of plants directly from transformed apical meristems.

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“…The applicability of the particle bombardment to introduce and transiently express genes into dry bean was demonstrated in the beginning of 1990s (Genga et al 1992;Aragão et al 1992Aragão et al , 1993. The bombardment of meristematic cells of embryonic axes revealed that foreign genes could efficiently reach the superficial cell layers, demonstrating the feasibility of producing transgenic bean using this method (Aragão et al 1993). Russell et al (1993) reported the production of transgenic navy bean plants using an electrical particle acceleration device.…”

Section: Transformationmentioning

confidence: 99%

Genomics of Phaseolus Beans, a Major Source of Dietary Protein and Micronutrients in the Tropics

Gepts

Aragão²,

Barros³

et al.

Plant Genetics and Genomics: Crops and Models

146

105

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except for brief excerpts in connection with reviews or scholarly analysis. Use in connection with any form of information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed is forbidden. The use in this publication of trade names, trademarks, service marks, and similar terms, even if they are not identified as such, is not to be taken as an expression of opinion as to whether or not they are subject to proprietary rights.Cover illustration: Small holders in tropics: Photograph by Gisela Orjeda, Bioversity International. Most of bananas grown in the tropics are produced by smallholders for home consumption or selling in local markets.

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Factors influencing transient gene expression in bean (Phaseolus vulgaris L.) using an electrical particle acceleration device

Cited by 39 publications

References 17 publications

Enhanced Transgene Expression in Sugarcane by Co-Expression of Virus-Encoded RNA Silencing Suppressors

Enhanced Transgene Expression in Sugarcane by Co-Expression of Virus-Encoded RNA Silencing Suppressors

Susceptibility of Common and Tepary Beans to Agrobacterium spp. Strains and Improvement of Agrobacterium-mediated Transformation Using Microprojectile Bombardment

Genomics of Phaseolus Beans, a Major Source of Dietary Protein and Micronutrients in the Tropics

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