The Casuarina glauca metallothionein I promoter in nodulated transgenic hairy roots of the actinorhizal plant Datisca glomerata

2011 ◽  
Vol 38 (9) ◽  
pp. 728 ◽  
Author(s):  
Behnoosh Rashidi ◽  
Sara Mehrabi ◽  
Kirill Demchenko ◽  
Katharina Pawlowski
Keyword(s):  
Plant Species ◽  
Hairy Roots ◽  
Lotus Japonicus ◽  
Root Systems ◽  
Root Cortex ◽  
Model Legume ◽  
Casuarina Glauca ◽  
Actinorhizal Plant ◽  
Hormone Balance ◽  
Nicotiana Tabacum L

The activity of the promoter of a metallothionein gene expressed in actinorhizal nodules of Casuarina glauca Sieber ex Spreng., CgMT1, has previously been analysed in Casaurinaceae and in tobacco (Nicotiana tabacum L.), Arabidopsis and rice. In all these plants, the promoter showed high activity in the root cortex and epidermis, making it a useful tool for the expression of transgenes. Therefore, its activity was now analysed in transgenic root systems of Datisca glomerata (C. Presl) Baill, an actinorhizal plant from a different phylogenetic group than C. glauca, using the same CgMT1::GUS fusion as in previous studies. However, in contrast with all other plant species examined previously, the CgMT1::GUS construct showed no activity at all in D. glomerata hairy roots: the expression pattern in nodules resembled that found in C. glauca nodules. This is probably due to the changed hormone balance in hairy roots since experiments on the CgMT1::GUS construct in transgenic Arabidopsis showed that CgMT1 promoter activity was repressed by auxin or cytokinin, respectively. Yet, in hairy roots of the model legume Lotus japonicus L. induced by the same Agrobacterium rhizogenes strain, the CgMT1 promoter was active in roots and not in nodules. These results indicate that although the expression of pRi T-DNA genes leads to changes in root hormone balance, these changes do not abolish the differences in phytohormone levels or sensitivity between plant species. Therefore, gene expression data obtained using transgenic hairy root systems have to be viewed with care, not only due to the disturbed hormone balance, but also because the effects of the pRI-T-DNA genes can differ between species.

scholarly journals Comparison of Nodule Induction in Legume and Actinorhizal Symbioses: The Induction of Actinorhizal Nodules Does Not Involve ENOD40

2003 ◽  
Vol 16 (9) ◽  
pp. 808-816 ◽  
Author(s):  
Carole Santi ◽  
Uritza von Groll ◽  
Ana Ribeiro ◽  
Maurizio Chiurazzi ◽  
Florence Auguy ◽  
...  
Keyword(s):  
Expression Pattern ◽  
Host Plants ◽  
Root Nodule ◽  
Higher Plants ◽  
Expression Patterns ◽  
Lotus Japonicus ◽  
Casuarina Glauca ◽  
Actinorhizal Plant ◽  
Allocasuarina Verticillata ◽  
Actinorhizal Nodules

Two types of root nodule symbioses are known for higher plants, legume and actinorhizal symbioses. In legume symbioses, bacterial signal factors induce the expression of ENOD40 genes. We isolated an ENOD40 promoter from an actinorhizal plant, Casuarina glauca, and compared its expression pattern in a legume (Lotus japonicus) and an actinorhizal plant (Allocasuarina verticillata) with that of an ENOD40 promoter from the legume soybean (GmENOD402). In the actinorhizal Allocasuarina sp., CgENOD40-GUS and GmENOD40-2-GUS showed similar expression patterns in both vegetative and symbiotic development, and neither promoter was active during nodule induction. The nonsymbiotic expression pattern of CgENOD40-GUS in the legume genus Lotus resembled the nonsymbiotic expression patterns of legume ENOD40 genes however, in contrast to GmENOD40-2-GUS, CgENOD40-GUS was not active during nodule induction. The fact that only legume, not actinorhizal, ENOD40 genes are induced during legume nodule induction can be linked to the phloem unloading mechanisms established in the zones of nodule induction in the roots of both types of host plants.

scholarly journals Analysis of Promoter Activity of the Early Nodulin Enod40 in Lotus japonicus

2005 ◽  
Vol 18 (5) ◽  
pp. 414-427 ◽  
Author(s):  
Mette Grønlund ◽  
Andreas Roussis ◽  
Emmanouil Flemetakis ◽  
Nicolette E. M. Quaedvlieg ◽  
Helmi R. M. Schlaman ◽  
...  
Keyword(s):  
Promoter Activity ◽  
Lotus Japonicus ◽  
Root Hairs ◽  
Early Time ◽  
Cellular Level ◽  
Nod Factors ◽  
Root Cortex ◽  
Model Legume ◽  
Early Nodulin ◽  
Nodulin Genes

Our comparative studies on the promoter (pr) activity of Enod40 in the model legume Lotus japonicus in stably transformed GusA reporter lines and in hairy roots of L. japonicus demonstrate a stringent regulation of the Enod40 promoter in the root cortex and root hairs in response to Nod factors. Interestingly, the L. japonicus Enod40-2 promoter fragment also shows symbiotic activity in the reverse orientation. Deletion analyses of the Glycine max (Gm) Enod40 promoter revealed the presence of a minimal region -185 bp upstream of the transcription start. Stable transgenic L. japonicus reporter lines were used in bioassays to test the effect of different compounds on early symbiotic signaling. The responses of prGmEnod40 reporter lines were compared with the responses of L. japonicus (Lj) reporter lines based on the LjNin promoter. Both reporter lines show very early activity postinoculation in root hairs of the responsive zone of the root and later in the dividing cells of nodule primordia. The LjNin promoter was found to be more responsive than the GmEnod40 promoter to Nod factors and related compounds. The use of prGmEnod40 reporter lines to analyze the effect of nodulin genes on the GmEnod40 promoter activity indicates that LJNIN has a positive effect on the regulation of the Enod40 promoter, whereas the latter is not influenced by ectopic overexpression of its own gene product. In addition to pointing to a difference in the regulation of the two nodulin genes Enod40 and Nin during early time points of symbiosis, the bioassays revealed a difference in the response to the synthetic cytokinin 6-benzylaminopurine (BAP) between alfalfa and clover and L. japonicus. In alfalfa and clover, Enod40 expression was induced upon BAP treatment, whereas this seems not to be the case in L. japonicus; these results correlate with effects at the cellular level because BAP can induce pseudonodules in alfalfa and clover but not in L. japonicus. In conclusion, we demonstrate the applicability of the described L. japonicus reporter lines in analyses of the specificity of compounds related to nodulation as well as for the dissection of the interplay between different nodulin genes.

scholarly journals Host preference and invasiveness of commensals in the Lotus and Arabidopsis root microbiota

2021 ◽  
Author(s):  
Kathrin Wippel ◽  
Ke Tao ◽  
Yulong Niu ◽  
Rafal Zgadzaj ◽  
Rui Guan ◽  
...  
Keyword(s):  
Plant Species ◽  
Host Preference ◽  
Lotus Japonicus ◽  
Culture Collection ◽  
Community Context ◽  
Commensal Bacteria ◽  
Priority Effects ◽  
Model Legume ◽  
Culture Independent ◽  
Root Microbiota

AbstractHealthy plants are colonized by microorganisms from the surrounding environment, which form stable communities and provide beneficial services to the host. Culture-independent profiling of the bacterial root microbiota shows that different plant species are colonized by distinct bacterial communities, even if they share the same habitat. It is, however, not known to what extent the host actively selects these communities and whether commensal bacteria are adapted to a specific plant species. Here, we report a sequence-indexed culture collection from roots and nodules of the model legume Lotus japonicus that contains representatives from the majority of species found in culture-independent profiles. Using taxonomically paired synthetic communities from L. japonicus and the crucifer Arabidopsis thaliana in a multi-species gnotobiotic system, we detect clear signatures of host preference among commensal bacteria in a community context, but not in mono-associations. Sequential inoculation of either host reveals strong priority effects during the assembly of the root microbiota, where established communities are resilient to invasion by late-comers. However, we found that host preference by commensal bacteria confers a competitive advantage in their native host. We reveal that host preference is prevalent in commensal bacteria from diverse taxonomic groups and that this trait is directly linked to their invasiveness into standing root-associated communities.

scholarly journals Gene Silencing by Expression of Hairpin RNA in Lotus japonicus Roots and Root Nodules

2003 ◽  
Vol 16 (8) ◽  
pp. 663-668 ◽  
Author(s):  
Hirotaka Kumagai ◽  
Hiroshi Kouchi
Keyword(s):  
Hairy Root ◽  
Hairy Roots ◽  
Rna Silencing ◽  
Root Nodules ◽  
Lotus Japonicus ◽  
Loss Of Function ◽  
Coding Region ◽  
Hairpin Rna ◽  
Model Legume ◽  
Root Transformation

We investigated the efficacy of self-complementary hairpin RNA (hpRNA) expression to induce RNA silencing in the roots and nodules of model legume Lotus japonicus, using hairy root transformation mediated by Agrobacterium rhizogenes. Transgenic lines that express β-glucuronidase (GUS) by constitutive or nodule-specific promoters were supertransformed by infection of A. rhizogenes harboring constructs for the expression of hpRNAs with sequences complementary to the GUS coding region. GUS activity in more than 60% of the hairy roots was decreased or silenced almost completely. Silencing of the GUS gene was also observed in symbiotic nodules formed on hairy roots in both early and late stages of nodule organogenesis. These results indicate that transient RNA silencing by hairy root transformation provides a powerful tool for loss-of-function analyses of genes that function in roots and root nodules.

scholarly journals Characterization and Expression Analysis of Genes Encoding Phosphoenolpyruvate Carboxylase and Phosphoenolpyruvate Carboxylase Kinase of Lotus japonicus, a Model Legume

2003 ◽  
Vol 16 (4) ◽  
pp. 281-288 ◽  
Author(s):  
Tomomi Nakagawa ◽  
Tomoko Izumi ◽  
Mari Banba ◽  
Yosuke Umehara ◽  
Hiroshi Kouchi ◽  
...  
Keyword(s):  
Phosphoenolpyruvate Carboxylase ◽  
Root Nodules ◽  
Expression Patterns ◽  
Phosphorylation Site ◽  
Lotus Japonicus ◽  
Specific Protein ◽  
Mrna Levels ◽  
Model Legume ◽  
Putative Phosphorylation Site

Phosphoenolpyruvate carboxylases (PEPCs), one form of which in each legume species plays a central role in the carbon metabolism in symbiotic root nodules, are activated through phosphorylation of a conserved residue by a specific protein kinase (PEPC-PK). We characterized the cDNAs for two PEPC isoforms of Lotus japonicus, an amide-translocating legume that forms determinate nodules. One gene encodes a nodule-enhanced form, which is more closely related to the PEPCs in amide-type indeterminate nodules than those in ureide-type determinate nodules. The other gene is expressed in shoots and roots at a low level. Both forms have the putative phosphorylation site, Ser11. We also isolated a cDNA and the corresponding genomic DNA for PEPC-PK of L. japonicus. The recombinant PEPC-PK protein expressed in Escherichia coli phosphorylated recombinant maize C4-form PEPC efficiently in vitro. The level of mRNA for PEPC-PK was high in root nodules, and those in shoots and roots were also significant. In situ hybridization revealed that the expression patterns of the transcripts for PEPC and PEPC-PK were similar in mature root nodules, but were different in emerging nodules. When L. japonicus seedlings were subjected to prolonged darkness and subsequent illumination, the activity of PEPC-PK and the mRNA levels of both PEPC and PEPC-PK in nodules decreased and then recovered, suggesting that they are regulated according to the amounts of photosynthates transported from shoots.

scholarly journals Αναπρογραμματισμός του μεταβολισμού του Lotus japonicus σε μεταγραφικό, βιοχημικό και μεταβολομικό επίπεδο κατά τη συμβιωτική αζωτοδέσμευση

2016 ◽  
Author(s):  
Χρυσάνθη Καλλονιάτη
Keyword(s):  
Nitrogen Fixation ◽  
Metabolite Profiling ◽  
Symbiotic Nitrogen Fixation ◽  
Sulfur Metabolism ◽  
Lotus Japonicus ◽  
Nitrogen Fixing ◽  
Model Legume ◽  
Whole Plant ◽  
Plant Level ◽  
Molecular And Biochemical Mechanisms

Symbiotic nitrogen fixation in legumes takes place in specialized organs called nodules,which become the main source of assimilated nitrogen for the whole plant. Symbiotic nitro‐gen fixation requires exquisite integration of plant and bacterial metabolism and involvesglobal changes in gene expression and metabolite accumulation in both rhizobia and thehost plant. In order to study the metabolic changes mediated by symbiotic nitrogen fixationon a whole‐plant level, metabolite levels were profiled by gas chromatography–mass spec‐trometry in nodules and non‐symbiotic organs of Lotus japonicus plants uninoculated or in‐oculated with M. loti wt,  ΔnifA or  ΔnifH fix‐ strains. Furthermore, transcriptomic andbiochemical approaches were combined to study sulfur metabolism in nodules, its link tosymbiotic nitrogen fixation, and the effect of nodules on whole‐plant sulfur partitioning andmetabolism. It is well established that nitrogen and sulfur (S) metabolism are tightly en‐twined and sulfur is required for symbiotic nitrogen fixation, however, little is known aboutthe molecular and biochemical mechanisms governing sulfur uptake and assimilation duringsymbiotic nitrogen fixation. Transcript profiling in Lotus japonicus was combined with quan‐tification of S‐metabolite contents and APR activity in nodules and in non‐symbiotic organsof plants uninoculated or inoculated with M. loti wt, ΔnifA or ΔnifH fix‐ strains. Moreover,sulfate uptake and its distribution into different plant organs were analyzed and 35S‐flux intodifferent S‐pools was monitored. Metabolite profiling revealed that symbiotic nitrogen fixa‐tion results in dramatic changes of many aspects of primary and secondary metabolism innodules which leads to global reprogramming of metabolism of the model legume on awhole‐plant level. Moreover, our data revealed that nitrogen fixing nodules represent athiol‐rich organ. Their high APR activity and 35S‐flux into cysteine and its metabolites in com‐bination with the transcriptional up‐regulation of several genes involved in sulfur assimila‐tion highlight the function of nodules as a new site of sulfur assimilation. The higher thiolcontent observed in non‐symbiotic organs of nitrogen fixing plants in comparison touninoculated plants cannot be attributed to local biosynthesis, indicating that nodules couldserve as a novel source of reduced sulfur for the plant, which triggers whole‐plant repro‐gramming of sulfur metabolism. Interestingly, the changes in metabolite profiling and theenhanced thiol biosynthesis in nodules and their impact on the whole‐plant sulfur, carbonand nitrogen economy are dampened in fix‐ plants, which in most respects metabolically re‐sembled uninoculated plants, indicating a strong interaction between nitrogen fixation andsulfur and carbon metabolism.

Recent Advances in the Molecular Genetics of The Model Legume Lotus japonicus

1997 ◽  
pp. 255-258
Author(s):  
Leif Schauser ◽  
Leszek Boron ◽  
Eloisa Pajuelo ◽  
Thomas Thykjær ◽  
Dorthe Danielsen ◽  
...  
Keyword(s):  
Molecular Genetics ◽  
Lotus Japonicus ◽  
Model Legume ◽  
Recent Advances

scholarly journals Chitotetraose activates the fungal-dependent endosymbiotic signaling pathway in actinorhizal plant species

PLoS ONE ◽  
2019 ◽  
Vol 14 (10) ◽  
pp. e0223149 ◽  
Author(s):  
Mireille Chabaud ◽  
Joëlle Fournier ◽  
Lukas Brichet ◽  
Iltaf Abdou-Pavy ◽  
Leandro Imanishi ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Plant Species ◽  
Actinorhizal Plant

scholarly journals Accumulation of and Response to Auxins in Roots and Nodules of the Actinorhizal Plant Datisca glomerata Compared to the Model Legume Medicago truncatula

2019 ◽  
Vol 10 ◽  
Author(s):  
Irina V. Demina ◽  
Pooja Jha Maity ◽  
Anurupa Nagchowdhury ◽  
Jason L. P. Ng ◽  
Eric van der Graaff ◽  
...  
Keyword(s):  
Medicago Truncatula ◽  
Model Legume ◽  
Actinorhizal Plant
Sign in / Sign up

Export Citation Format

Share Document