An Agrobacterium-mediated transient gene expression system for intact leaves

Kapila, Jyoti; Rycke, Riet De; Montagu, Marc Van; Angenon, Geert

doi:10.1016/s0168-9452(96)04541-4

Cited by 620 publications

(466 citation statements)

References 22 publications

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“…The resulting vectors, named pK7WG2.LHCSR1 and pK7WG2.LHCSR1 His tag, were used for stable (25) and transient (32,33) A. tumefaciens-mediated transformation of plants.…”

Section: Resultsmentioning

confidence: 99%

Heterologous Expression of Moss Light-harvesting Complex Stress-related 1 (LHCSR1), the Chlorophyll a-Xanthophyll Pigment-protein Complex Catalyzing Non-photochemical Quenching, in Nicotiana sp.

Pinnola

Ghin

Gecchele

et al. 2015

Journal of Biological Chemistry

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Background: LHCSR protein in algae and mosses is essential for NPQ. Results: Expression and characterization of Physcomitrella patens LHCSR1 protein upon heterologous expression in N. benthamiana and N. tabacum was obtained. Conclusion: LHCSR1 is the first member of LHC protein family lacking Chlorophyll b. It is active in NPQ. Significance: LHCSR1 isolation is crucial for the elucidation of the NPQ mechanism.

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“…The resulting vectors, named pK7WG2.LHCSR1 and pK7WG2.LHCSR1 His tag, were used for stable (25) and transient (32,33) A. tumefaciens-mediated transformation of plants.…”

Section: Resultsmentioning

confidence: 99%

Heterologous Expression of Moss Light-harvesting Complex Stress-related 1 (LHCSR1), the Chlorophyll a-Xanthophyll Pigment-protein Complex Catalyzing Non-photochemical Quenching, in Nicotiana sp.

Pinnola

Ghin

Gecchele

et al. 2015

Journal of Biological Chemistry

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“…In the present study, transformation frequency was highest when co-cultivation was performed at 24°C. Co-cultivation at low temperature has been reported in several instances Kapila et al 1997;Dillen et al 1997;Popelka et al 2006). It has also been found that the temperature dependence profile of T-DNA transfer does not parallel with that of vir gene induction, indicating that another factor possibly the formation of a conjugal pilus is causing temperature sensitivity of T-DNA transfer .…”

Section: Optimization Of Transformation Protocolmentioning

confidence: 99%

Development of an efficient in vitro plant regeneration system amenable to Agrobacterium- mediated transformation of a recalcitrant grain legume blackgram (Vigna mungo L. Hepper)

Sainger

Chaudhary

Dahiya

et al. 2015

Physiol Mol Biol Plants

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An efficient, rapid and direct multiple shoot regeneration system amenable to Agrobacterium-mediated transformation from primary leaf with intact petiole of blackgram (Vigna mungo) is established for the first time. The effect of the explant type and its age, type and concentration of cytokinin and auxin either alone or in combination and genotype on multiple shoot regeneration efficiency and frequency was optimized. The primary leaf explants with petiole excised from 4-day-old seedlings directly developed multiple shoots (an average of 10 shoots/ explant) from the cut ends of the petiole in 95 % of the cultures on MSB (MS salts and B 5 vitamins) medium containing 1.0 μM 6-benzylaminopurine. Elongated (2-3 cm) shoots were rooted on MSB medium with 2.5 μM indole-butyric acid and resulted plantlets were hardened and established in soil, where they resumed growth and reached maturity with normal seed set. The regenerated plants were morphologically similar to seed-raised plants and required 8 weeks time from initiation of culture to establish them in soil. The regeneration competent cells present at the cut ends of petiole are fully exposed and are, thus, easily accessible to Agrobacterium, making this plant regeneration protocol amenable for the production of transgenic plants. The protocol was further successfully used to develop fertile transgenic plants of blackgram using Agrobacterium tumefaciens strain EHA 105 carrying a binary vector pCAMBIA2301 that contains a neomycin phosphotransferase gene (nptII) and a β-glucuronidase (GUS) gene (uidA) interrupted with an intron.The presence and integration of transgenes in putative T 0 plants were confirmed by polymerase chain reaction (PCR) and Southern blot hybridization, respectively. The transgenes were inherited in Mendelian fashion in T 1 progeny and a transformation frequency of 1.3 % was obtained. This protocol can be effectively used for transferring new traits in blackgram and other legumes for their quantitative and qualitative improvements.

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“…Tobacco plants were agroin¢ltrated according to the protocol of Kapila et al [7]. Plant tissue was harvested for extraction 4 days after transformation as described in Merle et al [8].…”

Section: Agroin¢ltration Of Tobacco Leavesmentioning

confidence: 99%

“…Transcription of all constructs was driven from the enhanced 35S promoter of cauli£ower mosaic virus and transcription was terminated using the t35S termination sequence. An Agrobacterium-mediated transient system [7] was used to express s-rGL, r-rGL and v-rGL after vacuum in¢ltration of tobacco leaves. Extraction and puri¢cation of the rGLs were carried out 4 days after agroin¢ltration.…”

Section: Production Of S-rgl R-rgl and V-rgl In Tobacco Leavesmentioning

confidence: 99%

“…Our aim was to develop a rapid tobacco-based transient expression system that allows the validation of the best expression targeting hypothesis for a given candidate protein within the shortest possible time and without generation of stable transgenic plants. We have previously shown that an adaptation of the agrobacterium in¢ltration technique [7] allowed us to demonstrate the introduction of a post-translational modi¢cation pathway into tobacco cells by transient coexpression of animal-derived prolyl-hydroxylase and its substrate, collagen, to increase hydroxyprolin content and thereby thermal stability of the collagen triple helix [8]. Vaquero et al have demonstrated with a similar system that di¡erent expression approaches can be tested for recombinant antibodies [9].…”

Section: Introductionmentioning

confidence: 99%

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A transient tobacco expression system coupled to MALDI‐TOF‐MS allows validation of the impact of differential targeting on structure and activity of a recombinant therapeutic glycoprotein produced in plants

Mokrzycki-Issartel¹,

Bouchon

Farrer³

et al. 2003

FEBS Letters

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Tobacco-based transient expression was employed to elucidate the impact of di¡erential targeting to subcellular compartments on activity and quality of gastric lipase as a model for the production of recombinant glycoproteins in plants. Overall N-linked glycan structures of recombinant lipase were analyzed and for the ¢rst time sugar structures of its four individual N-glycosylation sites were determined in situ by matrix-assisted laser desorption/ionization time-of-£ight mass spectrometry (MALDI-TOF-MS) on a trypsin digest without isolation or deglycosylation of the peptides. Three glycosylation sites contain both complex-type N-glycans and high-mannose-type structures, the fourth is exclusively linked to high-mannose glycans. Although the overall pattern of glycan structures is in£uenced by the targeting, our results show that the type of glycans found linked to a given Asn residue is largely in£uenced by the physico-chemical environment of the site. The transient tobacco system combined with MALDI-TOF-MS appears to be a useful tool for the evaluation of glycoprotein production in plants. ß

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An Agrobacterium-mediated transient gene expression system for intact leaves

Cited by 620 publications

References 22 publications

Heterologous Expression of Moss Light-harvesting Complex Stress-related 1 (LHCSR1), the Chlorophyll a-Xanthophyll Pigment-protein Complex Catalyzing Non-photochemical Quenching, in Nicotiana sp.

Heterologous Expression of Moss Light-harvesting Complex Stress-related 1 (LHCSR1), the Chlorophyll a-Xanthophyll Pigment-protein Complex Catalyzing Non-photochemical Quenching, in Nicotiana sp.

Development of an efficient in vitro plant regeneration system amenable to Agrobacterium- mediated transformation of a recalcitrant grain legume blackgram (Vigna mungo L. Hepper)

A transient tobacco expression system coupled to MALDI‐TOF‐MS allows validation of the impact of differential targeting on structure and activity of a recombinant therapeutic glycoprotein produced in plants

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