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Tal Dahan-Meir, Shdema Filler-Hayut, Cathy Melamed-Bessudo, Samuel Bocobza, Henryk Czosnek, Asaph Aharoni, Avraham A Levy
Current breeding relies mostly on random mutagenesis and recombination to generate novel genetic variation. However, targeted genome editing is becoming an increasingly important tool for precise plant breeding. Using the CRISPR-Cas system combined with the bean yellow dwarf virus rolling circle replicon we optimized a method for targeted mutagenesis and gene replacement in tomato. The carotenoid isomerase (CRTISO) and phytoene synthase 1 (PSY1) genes from the carotenoid biosynthesis pathway were chosen as targets due to their easily detectable change of phenotype...
April 18, 2018: Plant Journal: for Cell and Molecular Biology
Thomas Cavalier-Smith, Ema E Chao, Rhodri Lewis
Infrakingdom Rhizaria is one of four major subgroups with distinct cell body plans that comprise eukaryotic kingdom Chromista. Unlike other chromists, Rhizaria are mostly heterotrophic flagellates, amoebae or amoeboflagellates, commonly with reticulose (net-like) or filose (thread-like) feeding pseudopodia; uniquely for eukaryotes, cilia have proximal ciliary transition-zone hub-lattices. They comprise predominantly flagellate phylum Cercozoa and reticulopodial phylum Retaria, whose exact phylogenetic relationship has been uncertain...
April 17, 2018: Protoplasma
Yu Song, Hang Zhang, Hongguang You, Yuanming Liu, Chao Chen, Xu Feng, Xingyu Yu, Shengyang Wu, Libo Wang, Shihua Zhong, Qiang Li, Yanming Zhu, Xiaodong Ding
The plant SnRK1 kinases play the central roles in the processes of energy balance, hormone perception, stress resistance, metabolism, growth and development. However, the functions of these kinases are still elusive. In this study, we used GsSnRK1 of wild soybean as bait to perform library-scale screens by the means of yeast two-hybrid to identify its interacting proteins. The putative interactions were verified by yeast re-transformation and β-galactodase assays, and the selected interactions were further confirmed in planta by BiFC and biochemical Co-IP assays...
April 17, 2018: Plant, Cell & Environment
Arne Weinhold, Elham Karimi Dorcheh, Ran Li, Natarajan Rameshkumar, Ian T Baldwin
Plant-microbe associations are thought to be beneficial for plant growth and resistance against biotic or abiotic stresses, but for natural ecosystems, the ecological analysis of microbiome function remains in its infancy. We used transformed wild tobacco plants ( Nicotiana attenuata ) which constitutively express an antimicrobial peptide (Mc-AMP1) of the common ice plant, to establish an ecological tool for plant-microbe studies in the field. Transgenic plants showed in planta activity against plant-beneficial bacteria and were phenotyped within the plants´ natural habitat regarding growth, fitness and the resistance against herbivores...
April 17, 2018: ELife
Jingfang Hao, Pierre Pétriacq, Linda de Bont, Michael Hodges, Bertrand Gakière
The flavoprotein l-aspartate oxidase (LASPO) is the first enzyme of the de novo biosynthetic pathway of NAD+ in plants. Although LASPO is considered pivotal to maintain NAD+ homeostasis, it has not been hitherto characterized in plants. Here, the cDNA encoding the LASPO from the model plant Arabidopsis thaliana (AtLASPO, At5g14760) has been cloned and expressed in Escherichia coli for subsequent enzyme characterization. The purified AtLASPO enzyme displayed a Km of 0.79 mM for l-aspartate and a kcat of 0...
June 2018: Plant Science: An International Journal of Experimental Plant Biology
Sunita A Ramesh, Muhammad Kamran, Wendy Sullivan, Larissa Chirkova, Mamoru Okamoto, Fien Degryse, Michael McLauchlin, Matthew Gilliham, Stephen D Tyerman
Plant aluminium-activated malate transporters (ALMTs) are currently classified as anion channels; they are also known to be regulated by diverse signals, leading to a range of physiological responses. Gamma-aminobutyric acid (GABA) regulation of anion flux through ALMT proteins requires a specific amino acid motif in ALMTs that shares similarity with a GABA-binding site in mammalian GABAA receptors. Here, we explore why TaALMT1-activation leads to a negative correlation between malate efflux and endogenous GABA concentrations ([GABA]i) in both wheat (Titicum aestivum L) root tips and in heterologous expression systems...
April 4, 2018: Plant Cell
Philippe V Jutras, Marie-Claire Goulet, Pierre-Olivier Lavoie, Marc-André D'Aoust, Frank Sainsbury, Dominique Michaud
Cellular engineering approaches have been proposed to mitigate unintended proteolysis in plant protein biofactories, involving the design of protease activity-depleted environments by gene silencing or in situ inactivation with accessory protease inhibitors. Here, we assessed the impact of Influenza virus M2 proton channel on host protease activities and recombinant protein processing in the cell secretory pathway of Nicotiana benthamiana leaves. Transient co-expression assays with M2 and GFP variant pHluorin were first conducted to illustrate the potential of proton export from the Golgi lumen to promote recombinant protein yield...
April 4, 2018: Plant Biotechnology Journal
Gavin Anderson, Chanyong Jang, Renyuan Wang, Michael Goodin
The ability of the matrix (M) protein of potato yellow dwarf virus (PYDV) to remodel nuclear membranes is controlled by a di-leucine motif located at residues 223 and 224 of its primary structure. This function can be uncoupled from that of its nuclear localization signal (NLS), which is controlled primarily by lysine and arginine residues immediately downstream of the LL motif. In planta localization of green fluorescent protein fusions, bimolecular fluorescence complementation assays with nuclear import receptor importin-α1 and yeast-based nuclear import assays provided three independent experimental approaches to validate the authenticity of the M-NLS...
April 4, 2018: Journal of General Virology
Piotr Rozpądek, Agnieszka M Domka, Michał Nosek, Rafał Ważny, Roman J Jędrzejczyk, Monika Wiciarz, Katarzyna Turnau
Over the last years the role of fungal endophytes in plant biology has been extensively studied. A number of species were shown to positively affect plant growth and fitness, thus attempts have been made to utilize these microorganisms in agriculture and phytoremediation. Plant-fungi symbiosis requires multiple metabolic adjustments of both of the interacting organisms. The mechanisms of these adaptations are mostly unknown, however, plant hormones seem to play a central role in this process. The plant hormone strigolactone (SL) was previously shown to activate hyphae branching of mycorrhizal fungi and to negatively affect pathogenic fungi growth...
2018: Frontiers in Microbiology
Shiv S Pandey, Sucheta Singh, Harshita Pandey, Madhumita Srivastava, Tania Ray, Sumit Soni, Alok Pandey, Karuna Shanker, C S Vivek Babu, Suchitra Banerjee, M M Gupta, Alok Kalra
Tissue specific biosynthesis of secondary metabolites is a distinguished feature of medicinal plants. Withania somnifera, source of pharmaceutically important withanolides biosynthesizes withaferin-A in leaves and withanolide-A in roots. To increase the in planta withanolides production, a sustainable approach needs to be explored. Here, we isolated endophytes from different parts of W. somnifera plants and their promising role in in planta withanolide biosynthesis was established in both in-vivo grown as well in in-vitro raised composite W...
April 3, 2018: Scientific Reports
Supaporn Baiya, Bancha Mahong, Sang-Kyu Lee, Jong-Seong Jeon, James R Ketudat Cairns
The glycoside hydrolase family 1 members Os4BGlu14, Os4BGlu16, and Os4BGlu18 were proposed to be rice monolignol β-glucosidases. In vitro studies demonstrated that the Os4BGlu16 and Os4BGlu18 hydrolyze the monolignol glucosides coniferin and syringin with high efficiency compared to other substrates. The replacement of the conserved catalytic acid/base glutamate residue by a nonionizable glutamine residue in Os4BGlu14 suggested that it may be inactive as a β-glucosidase. Here, we investigated the activities of Os4BGlu14, Os4BGlu16, and Os4BGlu18 in planta by recombinant expression of their genes in the Arabidopsis bglu45-2 (monolignol β-glucosidase) mutant and analysis of monolignol glucosides by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MSMS)...
March 28, 2018: Plant Physiology and Biochemistry: PPB
Krissana Boonpa, Supaluk Tantong, Kamonwan Weerawanich, Pawinee Panpetch, Onanong Pringsulaka, Yodying Yingchutrakul, Sittiruk Roytrakul, Supaart Sirikantaramas
According to an in silico analysis, OsGASR3 (LOC_Os03g55290) from rice (Oryza sativa L.) was predicted to be involved in plant defense mechanisms. A semi-quantitative reverse transcription polymerase chain reaction assay revealed that OsGASR3 is highly expressed in the inflorescences of Thai jasmine rice (O. sativa L. subsp. indica 'KDML 105'). To characterize the biological activity of OsGASR3, we produced an OsGASR3-glutathione S-transferase fusion protein in Escherichia coli Rosetta-gami (DE3) cells for a final purified recombinant OsGASR3 yield of 0...
March 27, 2018: Journal of Plant Physiology
Rainer Hedrich, Thomas D Mueller, Dirk Becker, Irene Marten
Plants and animals in endosomes operate TPC1/SV-type cation channels. Although all plant cells functionally express at least one TPC1 gene, and the encoded SV channel was already discovered in the plant vacuole membrane two decades ago, its biological function has remained enigmatic. Recently, the structure of a plant TPC1/SV channel protein was determined. Insights into the 3D topology has now guided site-directed mutation approaches, enabling SV channel structure-function analyses and shed new light on earlier findings...
March 31, 2018: Molecular Plant
Sivabalan Karthik, Gadamchetty Pavan, Selvam Sathish, Ramamoorthy Siva, Periyasamy Suresh Kumar, Markandan Manickavasagam
Agrobacterium infection and regeneration of the putatively transformed plant from the explant remains arduous for some crop species like peanut. Henceforth, a competent and reproducible in planta genetic transformation protocol is established for peanut cv. CO7 by standardizing various factors such as pre-culture duration, acetosyringone concentration, duration of co-cultivation, sonication and vacuum infiltration. In the present investigation, Agrobacterium tumefaciens strain EHA105 harboring the binary vector pCAMBIA1301- bar was used for transformation...
April 2018: 3 Biotech
Morgan K Halane, Sang Hee Kim, Benjamin J Spears, Christopher M Garner, Conner J Rogan, Elizabeth C Okafor, Jianbin Su, Saikat Bhattacharjee, Walter Gassmann
Bacterial effector proteins secreted into host plant cells manipulate those cells to the benefit of the pathogen, but effector-triggered immunity (ETI) occurs when effectors are recognized by host resistance proteins. The RPS4/RRS1 pair recognizes the Pseudomonas syringae pv. pisi effector AvrRps4. AvrRps4 is processed in planta into AvrRps4N (133 amino acids), homologous to the N-termini of other effectors including the native P. syringae pv. tomato strain DC3000 effector HopK1, and AvrRps4C (88 amino acids)...
March 30, 2018: PLoS Pathogens
Gracia Montilla-Bascón, Julien Mandon, Frans J M Harren, Luis A J Mur, Simona M Cristescu, Elena Prats
Despite the established importance of nitric oxide (NO) in many physiological and molecular processes in plants, most methods for quantifying NO are open to criticism This reflects the differing methods either lacking specificity or sensitivity, or even from an undue dependence of results on experimental conditions (i.e., chemical concentrations, pH, etc.). In this chapter we describe a protocol to measure gaseous NO produced by a biological sample using quantum cascade laser (QCL)-based spectroscopy. This technique is based on absorption of the laser light by the NO molecules which have been passed from a biological sample into an optical s cell that is equipped with two mirrors placed at both ends...
2018: Methods in Molecular Biology
Michaël Mulot, Baptiste Monsion, Sylvaine Boissinot, Maryam Rastegar, Sophie Meyer, Nicole Bochet, Véronique Brault
Aphid-transmitted plant viruses are a threat for major crops causing massive economic loss worldwide. Members in the Luteoviridae family are transmitted by aphids in a circulative and non-replicative mode. Virions are acquired by aphids when ingesting sap from infected plants and are transported through the gut and the accessory salivary gland (ASG) cells by a transcytosis mechanism relying on virus-specific receptors largely unknown. Once released into the salivary canal, virions are inoculated to plants, together with saliva, during a subsequent feeding...
2018: Frontiers in Microbiology
Andrea Chini, Alessio Cimmino, Marco Masi, Pierluigi Reveglia, Paola Nocera, Roberto Solano, Antonio Evidente
Jasmonates are signaling compounds that regulate plant responses to stress. Jasmonic acid (JA) is the direct precursor of the bioactive plant hormone JA-Ile, the ligand of the COI1-JAZ co-receptor complex. JA, its methyl ester and three furanonyl esters were recently isolated from the grapevine pathogen Lasiodiplodia mediterannea sp. The JA ester lasiojasmonate A (LasA) is the first reported naturally occurring JA-furanone, whose mode of action is currently unexplored. Here we show that LasA activates many JA-regulated responses in planta, including protein degradation, gene expression, and physiological processes...
March 24, 2018: Journal of Experimental Botany
Fraser Murphy, Qin He, Miles Armstrong, Licida M Giuliani, Petra C Boevink, Wei Zhang, Zhendong Tian, Paul Rj Birch, Eleanor M Gilroy
Plant pathogens deliver effectors to manipulate processes in their hosts, creating a suitable environment for invasion and proliferation. Yet, little is known about the host proteins that are targeted by effectors from filamentous pathogens. Here, we show that stable transgenic expression in potato (Solanum tuberosum) and transient expression in Nicotiana benthamiana of the Arg-any amino acid-Leu-Arg (RXLR) effector Pi17316 enhances leaf colonization by the late blight pathogen Phytophthora infestans. Expression of Pi17316 also attenuates cell death triggered by the pathogen-associated molecular pattern Infestin 1 (INF1), indicating that the effector suppresses pattern-triggered immunity...
March 27, 2018: Plant Physiology
Yanbiao Zhou, Cong Liu, Dongying Tang, Lu Yan, Dan Wang, Yuanzhu Yang, Jinshan Gui, Xiao-Ying Zhao, Laigeng Li, Xiao-Dan Tang, Feng Yu, Jiang-Lin Li, Lan-Lan Liu, Yonghua Zhu, Jianzhong Lin, Xuan-Ming Liu
Salt stress can significantly affect plant growth and agricultural productivity. Receptor-like kinases (RLKs) are believed to play essential roles in plant growth, development and responses to abiotic stresses. Here, we identify a receptor-like cytoplasmic kinase, salt tolerance receptor-like cytoplasmic kinase 1 (STRK1), from rice (Oryza sativa L.) that positively regulates salt and oxidative stress tolerance. Our results show that STRK1 anchors and interacts with CatC at the plasma membrane via palmitoylation...
March 26, 2018: Plant Cell
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