scholarly journals Identification of New Potential Regulators of the Medicago truncatula—Sinorhizobium meliloti Symbiosis Using a Large-Scale Suppression Subtractive Hybridization Approach

2007 ◽  
Vol 20 (3) ◽  
pp. 321-332 ◽  
Author(s):  
Laurence Godiard ◽  
Andreas Niebel ◽  
Fabienne Micheli ◽  
Jérôme Gouzy ◽  
Thomas Ott ◽  
...  
Keyword(s):  
Signal Transduction ◽  
Suppression Subtractive Hybridization ◽  
Medicago Truncatula ◽  
Large Scale ◽  
Sinorhizobium Meliloti ◽  
Zinc Finger Protein ◽  
Gene Clusters ◽  
Subtractive Hybridization ◽  
Symbiotic Interaction ◽  
Genes Encoding

We set up a large-scale suppression subtractive hybridization (SSH) approach to identify Medicago truncatula genes differentially expressed at different stages of the symbiotic interaction with Sinorhizobium meliloti, with a particular interest for regulatory genes. We constructed 7 SSH libraries covering successive stages from Nod factor signal transduction to S. meliloti infection, nodule organogenesis, and functioning. Over 26,000 clones were differentially screened by two rounds of macroarray hybridizations. In all, 3,340 clones, corresponding to genes whose expression was potentially affected, were selected, sequenced, and ordered into 2,107 tentative gene clusters, including 767 MtS clusters corresponding to new M. truncatula genes. In total, 52 genes encoding potential regulatory proteins, including transcription factors (TFs) and other elements of signal transduction cascades, were identified. The expression pattern of some of them was analyzed by quantitative reverse-transcription polymerase chain reaction in wild-type and in Nod¯ M. truncatula mutants blocked before or after S. meliloti infection. Three genes, coding for TFs of the bHLH and WRKY families and a C2H2 zinc-finger protein, respectively, were found to be upregulated, following S. meliloti inoculation, in the infection-defective mutant lin, whereas the bHLH gene also was expressed in the root-hair-curling mutant hcl. The potential role of these genes in early symbiotic steps is discussed.

scholarly journals Expression of Medicago truncatula Genes Responsive to Nitric Oxide in Pathogenic and Symbiotic Conditions

2008 ◽  
Vol 21 (6) ◽  
pp. 781-790 ◽  
Author(s):  
Alberto Ferrarini ◽  
Matteo De Stefano ◽  
Emmanuel Baudouin ◽  
Chiara Pucciariello ◽  
Annalisa Polverari ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Medicago Truncatula ◽  
Regulatory Networks ◽  
Sinorhizobium Meliloti ◽  
Biotic Interactions ◽  
Hypersensitive Reaction ◽  
Nodule Formation ◽  
Polymorphism Analysis ◽  
Symbiotic Interaction ◽  
Physiological Relevance

Nitric oxide (NO) is involved in diverse physiological processes in plants, including growth, development, response to pathogens, and interactions with beneficial microorganisms. In this work, a dedicated microarray representing the widest database available of NO-related transcripts in plants has been produced with 999 genes identified by a cDNA amplified fragment length polymorphism analysis as modulated in Medicago truncatula roots treated with two NO donors. The microarray then was used to monitor the expression of NO-responsive genes in M. truncatula during the incompatible interaction with the foliar pathogen Colletotrichum trifolii race 1 and during the symbiotic interaction with Sinorhizobium meliloti 1021. A wide modulation of NO-related genes has been detected during the hypersensitive reaction or during nodule formation and is discussed with special emphasis on the physiological relevance of these genes in the context of the two biotic interactions. This work clearly shows that NO-responsive genes behave differently depending on the plant organ and on the type of interaction, strengthening the need to consider regulatory networks, including different signaling molecules.

scholarly journals The Medicago truncatula N5 Gene Encoding a Root-Specific Lipid Transfer Protein Is Required for the Symbiotic Interaction with Sinorhizobium meliloti

2009 ◽  
Vol 22 (12) ◽  
pp. 1577-1587 ◽  
Author(s):  
Youry Pii ◽  
Alessandra Astegno ◽  
Elisa Peroni ◽  
Massimo Zaccardelli ◽  
Tiziana Pandolfini ◽  
...  
Keyword(s):  
Medicago Truncatula ◽  
Sinorhizobium Meliloti ◽  
Lipid Transfer Protein ◽  
Symbiotic Association ◽  
Lipid Transfer ◽  
Symbiotic Interaction ◽  
Transgenic Roots ◽  
Transfer Protein ◽  
Small Proteins

The Medicago truncatula N5 gene is induced in roots after Sinorhizobium meliloti infection and it codes for a putative lipid transfer protein (LTP), a family of plant small proteins capable of binding and transferring lipids between membranes in vitro. Various biological roles for plant LTP in vivo have been proposed, including defense against pathogens and modulation of plant development. The aim of this study was to shed light on the role of MtN5 in the symbiotic interaction between M. truncatula and S. meliloti. MtN5 cDNA was cloned and the mature MtN5 protein expressed in Escherichia coli. The lipid binding capacity and antimicrobial activity of the recombinant MtN5 protein were tested in vitro. MtN5 showed the capacity to bind lysophospholipids and to inhibit M. truncatula pathogens and symbiont growth in vitro. Furthermore, MtN5 was upregulated in roots after infection with either the fungal pathogen Fusarium semitectum or the symbiont S. meliloti. Upon S. meliloti infection, MtN5 was induced starting from 1 day after inoculation (dpi). It reached the highest concentration at 3 dpi and it was localized in the mature nodules. MtN5-silenced roots were impaired in nodulation, showing a 50% of reduction in the number of nodules compared with control roots. On the other hand, transgenic roots overexpressing MtN5 developed threefold more nodules with respect to control roots. Here, we demonstrate that MtN5 possesses biochemical features typical of LTP and that it is required for the successful symbiotic association between M. truncatula and S. meliloti.

scholarly journals Expression Dynamics of the Medicago truncatula Transcriptome during the Symbiotic Interaction with Sinorhizobium meliloti: Which Role for Nitric Oxide?

2012 ◽  
Vol 161 (1) ◽  
pp. 425-439 ◽  
Author(s):  
Alexandre Boscari ◽  
Jennifer del Giudice ◽  
Alberto Ferrarini ◽  
Luca Venturini ◽  
Anne-Lise Zaffini ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Medicago Truncatula ◽  
Sinorhizobium Meliloti ◽  
Symbiotic Interaction

scholarly journals Towards sex identification of Asian Palmyra palm (Borassus flabelliferL.) by DNA fingerprinting, suppression subtractive hybridization andde novotranscriptome sequencing

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e7268
Author(s):  
Kwanjai Pipatchartlearnwong ◽  
Piyada Juntawong ◽  
Passorn Wonnapinij ◽  
Somsak Apisitwanich ◽  
Supachai Vuttipongchaikij
Keyword(s):  
Dna Fingerprinting ◽  
Suppression Subtractive Hybridization ◽  
Transcriptome Sequencing ◽  
Large Scale ◽  
De Novo ◽  
Subtractive Hybridization ◽  
Pcr Analysis ◽  
Dna Fingerprints ◽  
Juvenile Period ◽  
Male And Female

BackgroundAsian Palmyra palm, the source of palm-sugar, is dioecious with a long juvenile period requiring at least 12 years to reach its maturity. To date, there is no reliable molecular marker for identifying sexes before the first bloom, limiting crop designs and utilization. We aimed to identify sex-linked markers for this palm using PCR-based DNA fingerprinting, suppression subtractive hybridization (SSH) and transcriptome sequencing.MethodsDNA fingerprints were generated between males and females based on RAPD, AFLP, SCoT, modified SCoT, ILP, and SSR techniques. Large-scale cloning and screening of SSH libraries andde novotranscriptome sequencing of male and female cDNA from inflorescences were performed to identify sex-specific genes for developing sex-linked markers.ResultsThrough extensive screening and re-testing of the DNA fingerprints (up to 1,204 primer pairs) and transcripts from SSH (>10,000 clones) and transcriptome data, however, no sex-linked marker was identified. Althoughde novotranscriptome sequencing of male and female inflorescences provided ∼32 million reads and 187,083 assembled transcripts, PCR analysis of selected sex-highly represented transcripts did not yield any sex-linked marker. This result may suggest the complexity and small sex-determining region of the Asian Palmyra palm. To this end, we provide the first global transcripts of male and female inflorescences of Asian Palmyra palm. Interestingly, sequence annotation revealed a large proportion of transcripts related to sucrose metabolism, which corresponds to the sucrose-rich sap produced in the inflorescences, and these transcripts will be useful for further understanding of sucrose production in sugar crop plants. Provided lists of sex-specific and differential-expressed transcripts would be beneficial to the further study of sexual development and sex-linked markers in palms and related species.

scholarly journals Involvement of Glutaredoxin and Thioredoxin Systems in the Nitrogen-Fixing Symbiosis between Legumes and Rhizobia

Antioxidants ◽  
2018 ◽  
Vol 7 (12) ◽  
pp. 182 ◽  
Author(s):  
Geneviève Alloing ◽  
Karine Mandon ◽  
Eric Boncompagni ◽  
Françoise Montrichard ◽  
Pierre Frendo
Keyword(s):  
Sinorhizobium Meliloti ◽  
Root Nodule ◽  
Redox Balance ◽  
Atmospheric Nitrogen ◽  
Nitrogen Fixing ◽  
Symbiotic Interaction ◽  
Bacterial Differentiation ◽  
Genes Encoding ◽  
Transcriptomics Data

Leguminous plants can form a symbiotic relationship with Rhizobium bacteria, during which plants provide bacteria with carbohydrates and an environment appropriate to their metabolism, in return for fixed atmospheric nitrogen. The symbiotic interaction leads to the formation of a new organ, the root nodule, where a coordinated differentiation of plant cells and bacteria occurs. The establishment and functioning of nitrogen-fixing symbiosis involves a redox control important for both the plant-bacteria crosstalk and the regulation of nodule metabolism. In this review, we discuss the involvement of thioredoxin and glutaredoxin systems in the two symbiotic partners during symbiosis. The crucial role of glutathione in redox balance and S-metabolism is presented. We also highlight the specific role of some thioredoxin and glutaredoxin systems in bacterial differentiation. Transcriptomics data concerning genes encoding components and targets of thioredoxin and glutaredoxin systems in connection with the developmental step of the nodule are also considered in the model system Medicago truncatula–Sinorhizobium meliloti.

Identification of stage-specifically expressed genes of Trichinella spiralis by suppression subtractive hybridization

Parasitology ◽  
2007 ◽  
Vol 134 (10) ◽  
pp. 1443-1455 ◽  
Author(s):  
M. Y. LIU ◽  
X. L. WANG ◽  
B. Q. FU ◽  
C. Y. LI ◽  
X. P. WU ◽  
...  
Keyword(s):  
Suppression Subtractive Hybridization ◽  
Serine Proteases ◽  
Zinc Finger Protein ◽  
Trichinella Spiralis ◽  
Gene Families ◽  
Subtractive Hybridization ◽  
Cdna Clones ◽  
Dnase Ii ◽  
Finger Protein ◽  
Old Adult

SUMMARYNewborn larvae (NBL) and adult (Ad) stage-specifically expressed genes or members of gene families of Trichinella spiralis were identified by suppression subtractive hybridization (SSH)†. Six cDNA clones were identified as NBL stage-specific, including 1 member of the T. spiralis gene family encoding glutamic acid-rich proteins, 2 clones encoding novel serine proteases, 2 closely related clones encoding proteins that are members of a deoxyribonuclease II (DNase II)-like family and 1 clone with no similarity to known genes. Four stage-specific clones encoding homologues of retinoid X receptor, caveolin, C2H2 type zinc finger protein and a putative protein with no homology to known sequences were obtained from 3-day-old adult worms. One gene specifically up-regulated in the 5-day-old adult worms encoding a putative cuticle collagen was also identified.

scholarly journals Fungal Elicitation of Signal Transduction-Related Plant Genes Precedes Mycorrhiza Establishment and Requires the dmi3 Gene in Medicago truncatula

2004 ◽  
Vol 17 (12) ◽  
pp. 1385-1393 ◽  
Author(s):  
Stéphanie Weidmann ◽  
Lisa Sanchez ◽  
Julie Descombin ◽  
Odile Chatagnier ◽  
Silvio Gianinazzi ◽  
...  
Keyword(s):  
Signal Transduction ◽  
Medicago Truncatula ◽  
Sinorhizobium Meliloti ◽  
Expressed Sequence Tag ◽  
Signal Transduction Pathway ◽  
Suppressive Subtractive Hybridization ◽  
Cell Contact ◽  
Transduction Pathway ◽  
Transcript Accumulation ◽  
Plant Genes

Suppressive subtractive hybridization and expressed sequence tag sequencing identified 29 plant genes which are upregulated during the appressorium stage of mycorrhiza establishment between Medicago truncatula J5 (Myc+) and Glomus mosseae. Eleven genes coding plant proteins with predicted functions in signal transduction, transcription, and translation were investigated in more detail for their relation to early events of symbiotic interactions. Expression profiling showed that the genes are activated not only from the appressorium stage up to the fully established symbiosis in the Myc+ genotype of M. truncatula, but also when the symbionts are not in direct cell contact, suggesting that diffusible fungal molecules (Myc factors) play a role in the induction of a signal-transduction pathway. Transcript accumulation in roots of a mycorrhiza-defective Myc- dmi3 mutant of M. truncatula is not modified by appressorium formation or diffusible fungal molecules, indicating that the signal transduction pathway is required for a successful G. mosseae-M. truncatula interaction leading to symbiosis development. The symbiotic nodulating bacterium Sinorhizobium meliloti does not activate the 11 genes, which supposes early discrimination by plant roots between the microbial symbionts.

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