Olive Mild Mosaic Virus Coat Protein and P6 Are Suppressors of RNA Silencing, and Their Silencing Confers Resistance against OMMV by Carla MR Varanda, Patrick Materatski, Maria Doroteia Campos, Maria Ivone E. Clara, Gustavo Nolasco, Maria do Rosário Félix

“…Two hairpin constructs targeting each suppressor were agroinfiltrated in N. benthamiana plants, which were then inoculated with OMMV RNA. …”
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Rescue of a Cilevirus from infectious cDNA clones by Mikhail Oliveira Leastro, Elliot Watanabe Kitajima, Vicente Pallás, Jesús Ángel Sánchez-Navarro

“…Detection of CiLV-C subgenomic RNAs (sgRNAs) from agroinfiltrated and mechanically inoculated leaves further confirmed the infectivity of the clones. …”
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Transient expression of homologous hairpin RNA interferes with PVY transmission by aphids by Tenllado Francisco, Díaz-Ruíz José, Martínez-García Belén, Vargas Marisol

“…A complete and specific interference with aphid transmission of PVY was achieved by inducers of RNA silencing, as demonstrated by specific siRNAs accumulation in agroinfiltrated tissues. To our knowledge, this is the first report of successful interference with non-persistent transmission of a plant virus using RNA interference.…”
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Molecular characterization and pathogenicity of an infectious cDNA clone of tomato brown rugose fruit virus by Ziyue Ma, Hui Zhang, Ming Ding, Zhongkai Zhang, Xiuling Yang, Xueping Zhou

“…Further experimental evidence demonstrated that the rod-shaped virions accumulating in agroinfiltrated plants are sap-transmissible. This is the first report on the construction of a biologically active, full-length infectious cDNA clone of ToBRFV. …”
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Identification and Characterization of a Novel Quanzhou Mulberry Virus from Mulberry (<i>Morus alba</i>) by Jia Wei, Lei Chen, Zilong Xu, Peigang Liu, Yan Zhu, Tianbao Lin, Lu Yang, Yuan Huang, Zhiqiang Lv

“…An infectious clone for QMV was generated and agroinfiltrated into <i>Nicotiana benthamiana</i> and mulberry, resulting in no visible disease symptoms. …”
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Chrysanthemum Chlorotic Mottle Viroid-Mediated Trafficking of Foreign mRNA into Chloroplasts by Eseul Baek, Minju Park, Ju-Yeon Yoon, Peter Palukaitis

“…Chrysanthemum chlorotic mottle viroid (CChMVd) fused to the leader sequence of a reporter gene (mRFP) expressed transiently in agroinfiltrated Nicotiana benthamiana, was used to show that CChMVd can traffic into chloroplasts, thought to be the site of its replication. …”
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Subfamily-specific fluorescent probes for cysteine proteases display dynamic protease activities during seed germination by Lu, H, Chandrasekar, B, Oeljeklaus, J, Misas-Villamil, J, Wang, Z, Shindo, T, Bogyo, M, Kaiser, M, van der Hoorn, R

“…We applied protease activity profiling with these new probes on Arabidopsis (Arabidopsis thaliana) protease knockout lines and agroinfiltrated leaves to identify the probe targets and on other plant species to demonstrate their broad applicability. …”

Hibiscus chlorotic ringspot virus Coat Protein Upregulates Sulfur Metabolism Genes for Enhanced Pathogen Defense by Ruimin Gao, Florence Kai Lin Ng, Peng Liu, Sek-Man Wong

“…In both Hibiscus chlorotic ringspot virus (HCRSV)-infected and HCRSV coat protein (CP) agroinfiltrated plant leaves, we showed that sulfur metabolism pathway related genes—namely, sulfite oxidase (SO), sulfite reductase, and adenosine 5′-phosphosulfate kinase—were upregulated. …”
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The proteome of Nicotiana benthamiana is shaped by extensive protein processing by Zheng, K, Lyu, JC, Thomas, EL, Schuster, M, Sanguankiattichai, N, Ninck, S, Kaschani, F, Kaiser, M, van der Hoorn, RAL

“…Summary: Processing by proteases irreversibly regulates the fate of plant proteins and hampers the production of recombinant proteins in plants, yet only few processing events have been described in agroinfiltrated Nicotiana benthamiana, which has emerged as the main transient protein expression platform in plant science and molecular pharming. …”

Functional characterization of a strong bi-directional constitutive plant promoter isolated from cotton leaf curl Burewala virus. by Zainul A Khan, Malik Z Abdin, Jawaid A Khan

“…Further, the expression of GFP was monitored in agroinfiltrated leaves of N. benthamiana, N. tabacum and cotton (Gossypium hirsutum) plants using confocal laser scanning microscopy. …”
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Activity-based proteomics reveals nine target proteases for the recombinant protein-stabilizing inhibitor SlCYS8 in Nicotiana benthamiana by Jutras, P, Grosse-Holz, F, Kaschani, F, Kaiser, M, Michaud, D, Van Der Hoorn, R

“…Here, we performed activity-based protein profiling to identify proteases that are inhibited by SlCYS8 in agroinfiltrated Nicotiana benthamiana. We discovered that SlCYS8 selectively suppresses papain-like cysteine protease (PLCP) activity in both apoplastic fluids and total leaf extracts, while not affecting vacuolar-processing enzyme and serine hydrolase activity. …”

Development of an Efficient Virus Induced Gene Silencing Strategy in the Non-Model Wild Ginger-Zingiber zerumbet and Investigation of Associated Proteome Changes. by Chidambareswaren Mahadevan, Abdul Jaleel, Lokesh Deb, George Thomas, Manjula Sakuntala

“…Barley stripe mosaic virus (BSMV) silencing vectors containing an endogenous phytoene desaturase (PDS) gene fragment were agroinfiltrated into young leaves of Z. zerumbet under controlled growth conditions to effect virus-induced gene silencing (VIGS). …”
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Isopeptide Bonding <i>In Planta</i> Allows Functionalization of Elongated Flexuous Proteinaceous Viral Nanoparticles, including Non-Viable Constructs by Other Means by Daniel A. Truchado, Sara Rincón, Lucía Zurita, Flora Sánchez, Fernando Ponz

“…Moreover, the isopeptide bond between SpyTag and SpyCatcher occurred in vivo in co-agroinfiltrated <i>Nicotiana benthamiana</i> plants. These results demonstrated that SpyTag/SpyCatcher improves TuMV functionalization compared with previous approaches, thus implying the expansion of the application of the technology to elongated flexuous VNPs.…”
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Inhibition of potato leafroll virus multiplication and systemic translocation by siRNA constructs against putative ATPase fold of movement protein by Priyanka Kumari, Jitesh Kumar, Ravi Ranjan Kumar, Mohammad Ansar, Kumari Rajani, Sunil Kumar, Tushar Ranjan

“…Khufri Ashoka. Potato plants agroinfiltrated with MP siRNA constructs exhibited no rolling symptoms upon PLRV infection, indicating that the silencing of MP gene expression is an efficient method for generating PLRV-resistant potato plants. …”
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Dimerization of the Vacuolar Receptors AtRMR1 and -2 from Arabidopsis thaliana Contributes to Their Localization in the trans-Golgi Network by Alessandro Occhialini, Guillaume Gouzerh, Gian-Pietro Di Sansebastiano, Jean-Marc Neuhaus

“…Our experiments with agroinfiltrated Nicotiana benthamiana leaves demonstrated that AtRMR1 was localized in the endoplasmic reticulum (ER), while AtRMR2 was targeted to the trans-Golgi network (TGN) due to the presence of a cytosolic 23-amino acid sequence linker. …”
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Structure-guided mutagenesis of the capsid protein indicates that a nanovirus requires assembled viral particles for systemic infection. by Stefano Trapani, Eijaz Ahmed Bhat, Michel Yvon, Joséphine Lai-Kee-Him, François Hoh, Marie-Stéphanie Vernerey, Elodie Pirolles, Mélia Bonnamy, Guy Schoehn, Jean-Louis Zeddam, Stéphane Blanc, Patrick Bron

“…Using structure-guided mutagenesis based on a 3.2 Å resolution cryogenic-electron-microscopy reconstruction of the FBNSV particles, we demonstrate that specific site-directed mutations preventing capsid formation systematically suppress FBNSV long-distance movement, and thus systemic infection of host plants, despite positive detection of the mutated coat protein when the corresponding segment is agroinfiltrated into plant leaves. These results strongly suggest that the viral genome does not propagate within the plant vascular system under the form of uncoated DNA molecules or DNA:coat-protein complexes, but rather moves long distance as assembled viral particles.…”
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Metabolic engineering to simultaneously activate anthocyanin and proanthocyanidin biosynthetic pathways in Nicotiana spp. by Sandra Fresquet-Corrales, Edelín Roque, Alejandro Sarrión-Perdigones, Maricruz Rochina, María P López-Gresa, Huertas M Díaz-Mula, José M Bellés, Francisco Tomás-Barberán, José P Beltrán, Luis A Cañas

“…Transient expression experiments in N. benthamiana showed the activation of the anthocyanin pathway producing a purple color in the agroinfiltrated leaves and also the effective production of 208.5 nmol (-) catechin/g FW and 228.5 nmol (-) epicatechin/g FW measured by the p-dimethylaminocinnamaldehyde (DMACA) method. …”
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Naturally occurring substitution of an amino acid in a plant virus gene-silencing suppressor enhances viral adaptation to increasing thermal stress. by Lina Cai, Mingqing Dang, Yawen Yang, Ruoxin Mei, Fan Li, Xiaorong Tao, Peter Palukaitis, Randy Beckett, W Allen Miller, Stewart M Gray, Yi Xu

“…A P247S amino acid residue substitution in P0 increased CYDV-RPV replication after expression in agroinfiltrated plant leaves and increased viral pathogenicity of P0 generated from the heterologous Potato virus X expression vector system. …”
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Chile (Capsicum annuum) plants transformed with the RB gene from Solanum bulbocastanum are resistant to Phytophthora capsici. by Suman Bagga, Yvonne Lucero, Kimberly Apodaca, Wathsala Rajapakse, Phillip Lujan, Jose Luis Ortega, Champa Sengupta-Gopalan

“…Notably, a broad host resistance gene RB from Solanum bulbocastanum has been shown to confer resistance to P. infestans in both S. tuberosum and S. lycopersicum. We agroinfiltrated the RB gene into the leaves of susceptible chile plants, demonstrating that the gene is also capable of lending resistance to P. capsici in chile. …”
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Plant-produced chimeric virus-like particles - a new generation vaccine against African horse sickness by Daria A. Rutkowska, Nobalanda B. Mokoena, Tsepo L. Tsekoa, Vusi S. Dibakwane, Martha M. O’Kennedy

“…Results In this study, we demonstrated that transient co-expression of the four AHSV capsid proteins in agroinfiltrated Nicotiana benthamiana dXT/FT plants not only allowed for the assembly of homogenous AHSV-1 VLPs but also single, double and triple chimeric VLPs, where one capsid protein originated from one AHS serotype and at least one other capsid protein originated from another AHS serotype. …”
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