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Longxiang Wang, Longlong Wang, Qian Tan, Qiuling Fan, Hui Zhu, Zonglie Hong, Zhongming Zhang, Deqiang Duanmu
The targeted genome editing technique, CRISPR/Cas9 system, has been widely used to modify genes of interest in a predictable and precise manner. In this study, we describe the CRISPR/Cas9-mediated efficient editing of representative SNF (symbiotic nitrogen fixation) related genes in the model legume Lotus japonicus via Agrobacterium-mediated stable or hairy root transformation. We first predicted nine endogenous U6 genes in Lotus and then demonstrated the efficacy of the LjU6-1 gene promoter in driving expression of single guide RNAs (sgRNAs) by using a split yellow fluorescence protein (YFP) reporter system to restore the fluorescence in Arabidopsis protoplasts...
2016: Frontiers in Plant Science
Sudip Saha, Anindita Paul, Laura Herring, Ayan Dutta, Avisek Bhattacharya, Sandip Samaddar, Michael B Goshe, Maitrayee DasGupta
Symbiosis receptor kinase (SYMRK) is indispensable for activation of root nodule symbiosis (RNS) at both epidermal and cortical levels and is functionally conserved in legumes. Previously, we reported SYMRK to be phosphorylated on "gatekeeper" Tyr both in vitro as well as in planta. Since gatekeeper phosphorylation was not necessary for activity, the significance remained elusive. Herein, we show that substituting gatekeeper with nonphosphorylatable residues like Phe or Ala significantly affected autophosphorylation on selected targets on activation segment/αEF and β3-αC loop of SYMRK...
May 2016: Plant Physiology
Sandrine Fabre, Djamel Gully, Arthur Poitout, Delphine Patrel, Jean-François Arrighi, Eric Giraud, Pierre Czernic, Fabienne Cartieaux
Nitrogen fixation in the legume-rhizobium symbiosis is a crucial area of research for more sustainable agriculture. Our knowledge of the plant cascade in response to the perception of bacterial Nod factors has increased in recent years. However, the discovery that Nod factors are not involved in the Aeschynomene-Bradyrhizobium spp. interaction suggests that alternative molecular dialogues may exist in the legume family. We evaluated the conservation of the signaling pathway common to other endosymbioses using three candidate genes: Ca(2+)/Calmodulin-Dependent Kinase (CCaMK), which plays a central role in cross signaling between nodule organogenesis and infection processes; and Symbiosis Receptor Kinase (SYMRK) and Histidine Kinase1 (HK1), which act upstream and downstream of CCaMK, respectively...
December 2015: Plant Physiology
Sudip Saha, Maitrayee DasGupta
Recently we reported that overexpression of intracellular kinase domain of Symbiosis Receptor Kinase (SYMRK-kd) hyperactivated spontaneous nodulation in Medicago truncatula indicating the importance of SYMRK ectodomain in restricting nodule number. To clarify whether sunn and sickle pathways were overcome by SYMRK-kd for hyperactivation of nodule organogenesis, we overexpressed SYMRK-kd in these mutants and analyzed for spontaneous nodulation in absence of rhizobia. Spontaneous nodulation in skl/SYMRK-kd roots was 2-fold higher than A17/SYMRK-kd roots indicating nodule organogenesis induced by SYMRK-kd to be ethylene sensitive...
2015: Plant Signaling & Behavior
Arief Indrasumunar, Julia Wilde, Satomi Hayashi, Dongxue Li, Peter M Gresshoff
Association between legumes and rhizobia results in the formation of root nodules, where symbiotic nitrogen fixation occurs. The early stages of this association involve a complex of signalling events between the host and microsymbiont. Several genes dealing with early signal transduction have been cloned, and one of them encodes the leucine-rich repeat (LRR) receptor kinase (SymRK; also termed NORK). The Symbiosis Receptor Kinase gene is required by legumes to establish a root endosymbiosis with Rhizobium bacteria as well as mycorrhizal fungi...
March 15, 2015: Journal of Plant Physiology
Martina Katharina Ried, Meritxell Antolín-Llovera, Martin Parniske
Symbiosis Receptor-like Kinase (SYMRK) is indispensable for the development of phosphate-acquiring arbuscular mycorrhiza (AM) as well as nitrogen-fixing root nodule symbiosis, but the mechanisms that discriminate between the two distinct symbiotic developmental fates have been enigmatic. In this study, we show that upon ectopic expression, the receptor-like kinase genes Nod Factor Receptor 1 (NFR1), NFR5, and SYMRK initiate spontaneous nodule organogenesis and nodulation-related gene expression in the absence of rhizobia...
2014: ELife
Meritxell Antolín-Llovera, Elena Kristin Petutsching, Martina Katharina Ried, Volker Lipka, Thorsten Nürnberger, Silke Robatzek, Martin Parniske
The decision between defence and symbiosis signalling in plants involves alternative and modular plasma membrane-localized receptor complexes. A critical step in their activation is ligand-induced homo- or hetero-oligomerization of leucine-rich repeat (LRR)- and/or lysin motif (LysM) receptor-like kinases (RLKs). In defence signalling, receptor complexes form upon binding of pathogen-associated molecular patterns (PAMPs), including the bacterial flagellin-derived peptide flg22, or chitin. Similar mechanisms are likely to operate during the perception of microbial symbiont-derived (lipo)-chitooligosaccharides...
December 2014: New Phytologist
Sudip Saha, Ayan Dutta, Avisek Bhattacharya, Maitrayee DasGupta
Symbiosis Receptor Kinase (SYMRK), a member of the Nod factor signaling pathway, is indispensible for both nodule organogenesis and intracellular colonization of symbionts in rhizobia-legume symbiosis. Here, we show that the intracellular kinase domain of a SYMRK (SYMRK-kd) but not its inactive or full-length version leads to hyperactivation of the nodule organogenic program in Medicago truncatula TR25 (symrk knockout mutant) in the absence of rhizobia. Spontaneous nodulation in TR25/SYMRK-kd was 6-fold higher than rhizobia-induced nodulation in TR25/SYMRK roots...
December 2014: Plant Physiology
Meritxell Antolín-Llovera, Martina K Ried, Martin Parniske
Plants form root symbioses with fungi and bacteria to improve their nutrient supply. SYMBIOSIS RECEPTOR-LIKE KINASE (SYMRK) is required for phosphate-acquiring arbuscular mycorrhiza, as well as for the nitrogen-fixing root nodule symbiosis of legumes and actinorhizal plants, but its precise function was completely unclear. Here we show that the extracytoplasmic region of SYMRK, which comprises three leucine-rich repeats (LRRs) and a malectin-like domain (MLD) related to a carbohydrate-binding protein from Xenopus laevis, is cleaved to release the MLD in the absence of symbiotic stimulation...
February 17, 2014: Current Biology: CB
Irina V Demina, Tomas Persson, Patricia Santos, Marian Plaszczyca, Katharina Pawlowski
Actinorhizal root nodule symbioses are very diverse, and the symbiosis of Datisca glomerata has previously been shown to have many unusual aspects. In order to gain molecular information on the infection mechanism, nodule development and nodule metabolism, we compared the transcriptomes of D. glomerata roots and nodules. Root and nodule libraries representing the 3'-ends of cDNAs were subjected to high-throughput parallel 454 sequencing. To identify the corresponding genes and to improve the assembly, Illumina sequencing of the nodule transcriptome was performed as well...
2013: PloS One
Chao Wang, Hui Zhu, Liping Jin, Tao Chen, Longxiang Wang, Heng Kang, Zonglie Hong, Zhongming Zhang
SymRK-interacting protein 1 (SIP1) has previously been shown to interact with the symbiosis receptor kinase, SymRK, in Lotus japonicus. A longer variant of the SIP1 transcript, SIP1L, was isolated and characterized. SIP1L contains an additional 17 amino acids that make its C-terminus a complete heat shock protein 20 (Hsp20)-like domain. In contrast to SIP1S, the longer splicing variant SIP1L could not interact with SymRK. Both SIP1L and SIP1S transcripts could be detected in developing nodules and other plant tissues, although the former was always more abundant than the latter...
May 2013: Plant Molecular Biology
Anita Sellstedt, Kerstin H Richau
Studies of nitrogen-fixing properties among the Gram-positive Actinobacteria revealed that some species of Arthrobacter, Agromyces, Corynebacterium, Mycobacterium, Micromonospora, Propionibacteria and Streptomyces have nitrogen-fixing capacity. This is also valid for Frankia that fix nitrogen both in free-living and in symbiotic conditions. Frankia symbiosis results from interaction between the Frankia bacteria and dicotyledonous plants, that is, actinorhiza. These plants, which are important in forestry and agroforestry, form, together with the legumes (Fabales), a single nitrogen-fixing clade...
May 2013: FEMS Microbiology Letters
Aswathy Nair, Sujata Bhargava
Comparison of the expression of 13 genes involved in arbuscular mycorrhizal (AM) symbiosis was performed in a wild type tomato (Solanum lycopersicum cv 76R) and its reduced mycorrhizal colonization mutant rmc in response to colonization with Glomus fasiculatum. Four defense-related genes were induced to a similar extent in the mutant and wild type AM colonized plants, indicating a systemic response to AM colonization. Genes related to nutrient exchange between the symbiont partners showed higher expression in the AM roots of wild type plants than the mutant plants, which correlated with their arbuscular frequency...
December 2012: Plant Signaling & Behavior
Songli Yuan, Hui Zhu, Honglan Gou, Weiwei Fu, Lijing Liu, Tao Chen, Danxia Ke, Heng Kang, Qi Xie, Zonglie Hong, Zhongming Zhang
The symbiosis receptor kinase (SymRK) is required for morphological changes of legume root hairs triggered by rhizobial infection. How protein turnover of SymRK is regulated and how the nodulation factor signals are transduced downstream of SymRK are not known. In this report, a SymRK-interacting E3 ubiquitin ligase (SIE3) was shown to bind and ubiquitinate SymRK. The SIE3-SymRK interaction and the ubiquitination of SymRK were shown to occur in vitro and in planta. SIE3 represents a new class of plant-specific E3 ligases that contain a unique pattern of the conserved CTLH (for C-terminal to LisH), CRA (for CT11-RanBPM), and RING (for Really Interesting New Gene) domains...
September 2012: Plant Physiology
Rosana Sánchez-López, David Jáuregui, Carmen Quinto
Symbiotic legume-rhizobia relationship leads to the formation of nitrogen-fixing nodules. Successful nodulation depends on the expression and cross-talk of a batttery of genes, among them SymRK (symbiosis receptor-like kinase), a leucine-rich repeat receptor-like kinase. SymRK is required for the rhizobia invasion of root hairs, as well as for the infection thread and symbiosome formation. Using immunolocalization and downregulation strategies we have recently provided evidence of a new function of PvSymRK in nodulation...
June 2012: Plant Signaling & Behavior
Griet Den Herder, Satoko Yoshida, Meritxell Antolín-Llovera, Martina K Ried, Martin Parniske
The Lotus japonicus SYMBIOSIS RECEPTOR-LIKE KINASE (SYMRK) is required for symbiotic signal transduction upon stimulation of root cells by microbial signaling molecules. Here, we identified members of the SEVEN IN ABSENTIA (SINA) E3 ubiquitin-ligase family as SYMRK interactors and confirmed their predicted ubiquitin-ligase activity. In Nicotiana benthamiana leaves, SYMRK-yellow fluorescent protein was localized at the plasma membrane, and interaction with SINAs, as determined by bimolecular fluorescence complementation, was observed in small punctae at the cytosolic interface of the plasma membrane...
April 2012: Plant Cell
Tao Chen, Hui Zhu, Danxia Ke, Kai Cai, Chao Wang, Honglan Gou, Zonglie Hong, Zhongming Zhang
The symbiosis receptor kinase, SymRK, is required for root nodule development. A SymRK-interacting protein (SIP2) was found to form protein complex with SymRK in vitro and in planta. The interaction between SymRK and SIP2 is conserved in legumes. The SIP2 gene was expressed in all Lotus japonicus tissues examined. SIP2 represents a typical plant mitogen-activated protein kinase kinase (MAPKK) and exhibited autophosphorylation and transphosphorylation activities. Recombinant SIP2 protein could phosphorylate casein and the Arabidopsis thaliana MAP kinase MPK6...
February 2012: Plant Cell
Rosana Sánchez-López, David Jáuregui, Noreide Nava, Xóchitl Alvarado-Affantranger, Jesús Montiel, Olivia Santana, Federico Sanchez, Carmen Quinto
The symbiotic interaction of legumes and rhizobia results in the formation of nitrogen-fixing nodules. Nodulation depends on the finely coordinated expression of a battery of genes involved in the infection and the organogenesis processes. After Nod factor perception, symbiosis receptor kinase (SymRK) receptor triggers a signal transduction cascade essential for nodulation leading to cortical cell divisions, infection thread (IT) formation and final release of rhizobia to the intracellular space, forming the symbiosome...
December 2011: Plant, Cell & Environment
Sonja Kosuta, Mark Held, Md Shakhawat Hossain, Giulia Morieri, Amanda Macgillivary, Christopher Johansen, Meritxell Antolín-Llovera, Martin Parniske, Giles E D Oldroyd, Allan J Downie, Bogumil Karas, Krzysztof Szczyglowski
SYMRK is a leucine-rich-repeat (LRR)-receptor kinase that mediates intracellular symbioses of legumes with rhizobia and arbuscular mycorrhizal fungi. It participates in signalling events that lead to epidermal calcium spiking, an early cellular response that is typically considered as central for intracellular accommodation and nodule organogenesis. Here, we describe the Lotus japonicus symRK-14 mutation that alters a conserved GDPC amino-acid sequence in the SYMRK extracellular domain. Normal infection of the epidermis by fungal or bacterial symbionts was aborted in symRK-14...
September 2011: Plant Journal: for Cell and Molecular Biology
Frédéric Mahé, Dragomira Markova, Rémy Pasquet, Marie-Thérèse Misset, Abdelkader Aïnouche
SymRK is one of the key genes involved in initial steps of legume symbiotic association with fungi (mycorrhization) and nitrogen-fixing bacteria (nodulation). A large portion of the sequence encoding the extracellular domain of SYMRK was obtained for 38 lupine accessions and 2 outgroups in order to characterize this region, to evaluate its phylogenetic utility, and to examine whether its molecular evolutionary pattern is correlated with rhizobial diversity and specificity in Lupinus. The data suggested that, in Lupinus, SymRK is a single copy gene that shows good phylogenetic potential...
July 2011: Molecular Phylogenetics and Evolution
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