Transient plant transformation mediated by Agrobacterium tumefaciens: Principles, methods and applications

Křenek, Pavel; Šamajová, Olga; Luptovčiak, Ivan; Doskočilová, Anna; Komis, George; Šamaj, Jozef

doi:10.1016/j.biotechadv.2015.03.012

Cited by 184 publications

(120 citation statements)

References 249 publications

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“…Chinese hamster ovary cells remain the preferred host for the commercial production of monoclonal IgGs but alternative expression systems such as those involving transient expression in agroinfiltrated plants have attracted special attention in recent years [44]. A practical challenge at present to further confirm the potential of plants as bio-factories for therapeutic IgGs is to improve their overall stability by a better control of endogenous proteolysis in host tissues.…”

Section: Discussionmentioning

confidence: 99%

An Accessory Protease Inhibitor to Increase the Yield and Quality of a Tumour-Targeting mAb in Nicotiana benthamiana Leaves

et al. 2016

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The overall quality of recombinant IgG antibodies in plants is dramatically compromised by host endogenous proteases. Different approaches have been developed to reduce the impact of endogenous proteolysis on IgGs, notably involving site-directed mutagenesis to eliminate protease-susceptible sites or the in situ mitigation of host protease activities to minimize antibody processing in the cell secretory pathway. We here characterized the degradation profile of H10, a human tumour-targeting monoclonal IgG, in leaves of Nicotiana benthamiana also expressing the human serine protease inhibitor α1-antichymotrypsin or the cysteine protease inhibitor tomato cystatin SlCYS8. Leaf extracts revealed consistent fragmentation patterns for the recombinant antibody regardless of leaf age and a strong protective effect of SlCYS8 in specific regions of the heavy chain domains. As shown using an antigen-binding ELISA and LC-MS/MS analysis of antibody fragments, SlCYS8 had positive effects on both the amount of fully-assembled antibody purified from leaf tissue and the stability of biologically active antibody fragments containing the heavy chain Fc domain. Our data confirm the potential of Cys protease inhibitors as convenient antibody-stabilizing expression partners to increase the quality of therapeutic antibodies in plant protein biofactories.

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Section: Discussionmentioning

confidence: 99%

An Accessory Protease Inhibitor to Increase the Yield and Quality of a Tumour-Targeting mAb in Nicotiana benthamiana Leaves

et al. 2016

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“…Although Agrobacterium -mediated transient expression of genes into leaves of entire plants has been described for many plants, such a technique did not exist for sunflower, and in the Asteraceae family, it was described for lettuce (Krenek et al, 2015). Stable transformation of sunflower by A. tumefaciens is still difficult, with poor efficiency and is restricted to specific sunflower lines (Knittel et al, 1994; Malone-Schoneberg et al, 1994; Burrus et al, 1996; Lucas et al, 2000).…”

Section: Discussionmentioning

confidence: 99%

RXLR and CRN Effectors from the Sunflower Downy Mildew Pathogen Plasmopara halstedii Induce Hypersensitive-Like Responses in Resistant Sunflower Lines

et al. 2016

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Plasmopara halstedii is an obligate biotrophic oomycete causing downy mildew disease on sunflower, Helianthus annuus, an economically important oil crop. Severe symptoms of the disease (e.g., plant dwarfism, leaf bleaching, sporulation and production of infertile flower) strongly impair seed yield. Pl resistance genes conferring resistance to specific P. halstedii pathotypes were located on sunflower genetic map but yet not cloned. They are present in cultivated lines to protect them against downy mildew disease. Among the 16 different P. halstedii pathotypes recorded in France, pathotype 710 is frequently found, and therefore continuously controlled in sunflower by different Pl genes. High-throughput sequencing of cDNA from P. halstedii led us to identify potential effectors with the characteristic RXLR or CRN motifs described in other oomycetes. Expression of six P. halstedii putative effectors, five RXLR and one CRN, was analyzed by qRT-PCR in pathogen spores and in the pathogen infecting sunflower leaves and selected for functional analyses. We developed a new method for transient expression in sunflower plant leaves and showed for the first time subcellular localization of P. halstedii effectors fused to a fluorescent protein in sunflower leaf cells. Overexpression of the CRN and of 3 RXLR effectors induced hypersensitive-like cell death reactions in some sunflower near-isogenic lines resistant to pathotype 710 and not in susceptible corresponding lines, suggesting they could be involved in Pl loci-mediated resistances.

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“…Hypervirulent strains of Agrobacterium, such as the succinamopine-type EHA105 and AGL1, improve transformation efficiencies (Hellens et al, 2000;Zhi et al, 2015), and might increase agro-inoculation success rates. The chrysopinetype strain Chry5 was originally isolated from chrysanthemum (Shao et al, 2018) and its derivative strain CryX was reported to have agro-inoculation efficiencies 100-1000 times higher than those of commonly used Agrobacterium strains ( (Azhakanandam et al, 2007;Giritch et al, 2013;Gleba et al, 2014;Grimsley et al, 1986;Hahn et al, 2015;Krenek et al, 2015). Viruses might be asymptomatic, vertically inherited in host plants and lack cell-to-cell movement (Roossinck, 2010); these aspects should be taken into account during experimental design and result evaluation.…”

Section: and Inoculation Efficiencymentioning

confidence: 99%

Harnessed viruses in the age of metagenomics and synthetic biology: an update on infectious clone assembly and biotechnologies of plant viruses

Pasin

Menzel

Daròs

2019

Plant Biotechnology Journal

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Summary Recent metagenomic studies have provided an unprecedented wealth of data, which are revolutionizing our understanding of virus diversity. A redrawn landscape highlights viruses as active players in the phytobiome, and surveys have uncovered their positive roles in environmental stress tolerance of plants. Viral infectious clones are key tools for functional characterization of known and newly identified viruses. Knowledge of viruses and their components has been instrumental for the development of modern plant molecular biology and biotechnology. In this review, we provide extensive guidelines built on current synthetic biology advances that streamline infectious clone assembly, thus lessening a major technical constraint of plant virology. The focus is on generation of infectious clones in binary T‐ DNA vectors, which are delivered efficiently to plants by Agrobacterium . We then summarize recent applications of plant viruses and explore emerging trends in microbiology, bacterial and human virology that, once translated to plant virology, could lead to the development of virus‐based gene therapies for ad hoc engineering of plant traits. The systematic characterization of plant virus roles in the phytobiome and next‐generation virus‐based tools will be indispensable landmarks in the synthetic biology roadmap to better crops.

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Transient plant transformation mediated by Agrobacterium tumefaciens: Principles, methods and applications

Cited by 184 publications

References 249 publications

An Accessory Protease Inhibitor to Increase the Yield and Quality of a Tumour-Targeting mAb in Nicotiana benthamiana Leaves

An Accessory Protease Inhibitor to Increase the Yield and Quality of a Tumour-Targeting mAb in Nicotiana benthamiana Leaves

RXLR and CRN Effectors from the Sunflower Downy Mildew Pathogen Plasmopara halstedii Induce Hypersensitive-Like Responses in Resistant Sunflower Lines

Harnessed viruses in the age of metagenomics and synthetic biology: an update on infectious clone assembly and biotechnologies of plant viruses

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