scholarly journals Lotus japonicus Gene Ljsbp Is Highly Conserved Among Plants and Animals and Encodes a Homologue to the Mammalian Selenium-Binding Proteins

2002 ◽  
Vol 15 (4) ◽  
pp. 313-322 ◽  
Author(s):  
Emmanouil Flemetakis ◽  
Adamantia Agalou ◽  
Nektarios Kavroulakis ◽  
Maria Dimou ◽  
Anna Martsikovskaya ◽  
...  
Keyword(s):  
Polyclonal Antibodies ◽  
Lotus Japonicus ◽  
Root Hairs ◽  
Physiological Role ◽  
Cdna Clones ◽  
Vascular Bundles ◽  
Root Tips ◽  
Root Epidermis ◽  
Infected Cells ◽  
Low Levels

We have isolated and characterized a Lotus japonicus gene (Ljsbp) encoding a putative polypeptide with striking homology to the mammalian 56-kDa selenium-binding protein (SBP). cDNA clones homologous to LjSBP were also isolated from soybean, Medicago sativa, and Arabidopsis thaliana. Comparative expression studies in L. japonicus and A. thaliana showed that sbp transcripts are present in various tissues and at different levels. Especially in L. japonicus nodules and seedpods and A. thaliana siliques, sbp expression appears to be developmentally up-regulated. sbp Gene transcripts were localized by in situ hybridization in the infected cells and vascular bundles of young nodules, while in mature nodules, low levels of expression were only detected in the parenchymatous cells. Expression of sbp transcripts in young seedpods and siliques was clearly visible in vascular tissues and embryos, while in embryos, low levels of expression were detected in the root epidermis and the vascular bundles. Polyclonal antibodies raised against a truncated LjSBP recombinant protein recognized a poly-peptide of about 60 kDa in nodule extracts. Immunohistochemical experiments showed that accumulation of LjSBP occurred in root hairs, in the root epidermis above the nodule primordium, in the phloem of the vasculature, and abundantly in the infected cells of young nodules. Irrespective of the presence of rhizobia, expression of SBP was also observed in root tips, where it was confined in the root epidermis and protophloem cells. We hypothesize that LjSBP may have more than one physiological role and can be implicated in controlling the oxidation/reduction status of target proteins, in vesicular Golgi transport, or both.

scholarly journals Expression Analysis of PIN Genes in Root Tips and Nodules of Lotus japonicus

2019 ◽  
Vol 20 (2) ◽  
pp. 235 ◽  
Author(s):  
Izabela Sańko-Sawczenko ◽  
Dominika Dmitruk ◽  
Barbara Łotocka ◽  
Elżbieta Różańska ◽  
Weronika Czarnocka
Keyword(s):  
Root Nodule ◽  
Root Nodules ◽  
Quantitative Polymerase Chain Reaction ◽  
Lotus Japonicus ◽  
Nodule Development ◽  
Vascular Bundles ◽  
Root Tips ◽  
Gus Reporter ◽  
Gene Assay ◽  
Development And Differentiation

Auxins are postulated to be one of the pivotal factors in nodulation. However, their transporters in Lotus japonicus, the model species for the study of the development of determinate-type root nodules, have been scarcely described so far, and thus their role in nodulation has remained unknown. Our research is the first focusing on polar auxin transporters in L. japonicus. We analyzed and compared expression of PINs in 20 days post rhizobial inoculation (dpi) and 54 dpi root nodules of L. japonicus by real-time quantitative polymerase chain reaction (qPCR) along with the histochemical β-glucuronidase (GUS) reporter gene assay in transgenic hairy roots. The results indicate that LjPINs are essential during root nodule development since they are predominantly expressed in the primordia and young, developing nodules. However, along with differentiation, expression levels of several PINs decreased and occurred particularly in the nodule vascular bundles, especially in connection with the root’s stele. Moreover, our study demonstrated the importance of both polar auxin transport and auxin intracellular homeostasis during L. japonicus root nodule development and differentiation.

scholarly journals Supra-physiological levels of Gibberellins/DELLAs alter the patterning, morphology and abundance of root hairs in root tips of A. thaliana seedlings

2021 ◽  
Author(s):  
Iva McCarthy-Suarez
Keyword(s):  
Gene Expression ◽  
Cell Fate ◽  
Epidermal Cell ◽  
Root Hair ◽  
Root Hairs ◽  
Root Tip ◽  
Spatial Patterning ◽  
Root Tips ◽  
Root Epidermis

In spite of the known role of gibberellins (GAs), and of their antagonistic proteins, the DELLAs, in leaf hair production, no investigations, however, have assessed their hypothetical function in the production of root hairs. To this aim, the effects of supra-physiological levels of GAs/DELLAs on the spatial patterning of gene expression of the root hair (CPC) and root non-hair (GL2, EGL3 and WER) epidermal cell fate markers, as well as on the distribution, morphology and abundance of root hairs, were studied in root tips of 5-day-old A. thaliana seedlings. Results showed that excessive levels of GAs/DELLAs impaired the spatial patterning of gene expression of the root hair/non-hair epidermal cell fate markers, as well as the arrangement, shape and frequency of root hairs, giving rise to ectopic hairs and ectopic non-hairs, two-haired cells, two-tipped hairs, branched hairs, longer and denser hairs near the root tip under excessive DELLAs, and shorter and scarcer hairs near the root tip under excessive GAs. However, when the gai-1 (GA-insensitive-1) DELLA mutant protein was specifically over-expressed at the root epidermis, no changes in the patterning or abundance of root hairs occurred. Thus, these results suggest that, in seedlings of A. thaliana, the GAs/DELLAs might have a role in regulating the patterning, morphology and abundance of root hairs by acting from the sub-epidermal tissues of the root.

scholarly journals A Dicarboxylate Transporter, LjALMT4, Mainly Expressed in Nodules of Lotus japonicus

2016 ◽  
Vol 29 (7) ◽  
pp. 584-592 ◽  
Author(s):  
Kojiro Takanashi ◽  
Takayuki Sasaki ◽  
Tomohiro Kan ◽  
Yuka Saida ◽  
Akifumi Sugiyama ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Symbiotic Nitrogen Fixation ◽  
Lotus Japonicus ◽  
Specific Gene ◽  
Vascular Bundles ◽  
Transport Activity ◽  
Organ Specific ◽  
Infected Cells ◽  
Photosynthetic Products ◽  
Strong Candidate

Legume plants can establish symbiosis with soil bacteria called rhizobia to obtain nitrogen as a nutrient directly from atmospheric N2 via symbiotic nitrogen fixation. Legumes and rhizobia form nodules, symbiotic organs in which fixed-nitrogen and photosynthetic products are exchanged between rhizobia and plant cells. The photosynthetic products supplied to rhizobia are thought to be dicarboxylates but little is known about the movement of dicarboxylates in the nodules. In terms of dicarboxylate transporters, an aluminum-activated malate transporter (ALMT) family is a strong candidate responsible for the membrane transport of carboxylates in nodules. Among the seven ALMT genes in the Lotus japonicus genome, only one, LjALMT4, shows a high expression in the nodules. LjALMT4 showed transport activity in a Xenopus oocyte system, with LjALMT4 mediating the efflux of dicarboxylates including malate, succinate, and fumarate, but not tricarboxylates such as citrate. LjALMT4 also mediated the influx of several inorganic anions. Organ-specific gene expression analysis showed LjALMT4 mRNA mainly in the parenchyma cells of nodule vascular bundles. These results suggest that LjALMT4 may not be involved in the direct supply of dicarboxylates to rhizobia in infected cells but is responsible for supplying malate as well as several anions necessary for symbiotic nitrogen fixation, via nodule vasculatures.

scholarly journals Supra-physiological levels of gibberellins/DELLAs modify the root cell size/number and the root architecture in root tips of A. thaliana seedlings. Connections to the root hair patterning and abundance

2021 ◽  
Author(s):  
Iva McCarthy-Suarez
Keyword(s):  
Cell Size ◽  
Root Hair ◽  
Root Architecture ◽  
Root Hairs ◽  
Root Tip ◽  
Root Tips ◽  
Root Epidermis ◽  
Size Number ◽  
Tip Cells ◽  
Root Tip Cells

A previous study (McCarthy-Suarez, 2021) showed that growing A. thaliana seedlings for 5 days under excessive levels of gibberellins (GAs)/DELLAs altered the arrangement, shape and frequency of root hairs in root tips. Because no changes in the distribution or number of root hairs occurred when the gai-1 (gibberellin-insensitive-1) DELLA was over-expressed at the root epidermis, it was concluded that the GAs/DELLAs might regulate the root hair patterning and abundance in A. thaliana seedlings by acting from the root sub-epidermal tissues. In the present study, microscopy analyses showed that excessive levels of GAs/DELLAs also modified the size and number of root tip cells in A. thaliana seedlings. While excessive DELLAs shortened and widened the root epidermal, cortical, endodermal and pericycle cells, excessive GAs, excepting the epidermal cells, generally narrowed them. However, no changes of root cell size occurred when gai-1 was over-expressed at the root epidermis. In addition, high levels of DELLAs often induced extra cells at the root epidermis, cortex, endodermis and pericycle, whereas high levels of GAs sometimes induced extra cells at the root cortex and pericycle. On the other hand, excessive levels of DELLAs enhanced the outgrowth of lateral roots in root tips, unlike excessive levels of GAs. Thus, the results of this study suggest that supra-physiological levels of GAs/DELLAs might modify the size/number of root tip cells by acting from the root sub-epidermal tissues. This, in turn, might impact on the patterning and abundance of root hairs and on the root architecture.

scholarly journals A Dihydroflavonol-4-Reductase-Like Protein Interacts with NFR5 and Regulates Rhizobial Infection in Lotus japonicus

2019 ◽  
Vol 32 (4) ◽  
pp. 401-412 ◽  
Author(s):  
Liujian Duan ◽  
Junqing Pei ◽  
Yaping Ren ◽  
Hao Li ◽  
Xiangzhen Zhou ◽  
...  
Keyword(s):  
Lotus Japonicus ◽  
Root Hairs ◽  
Root Tips ◽  
Symbiotic Interaction ◽  
Nod Factor ◽  
Symbiotic Interactions ◽  
Infection Threads ◽  
Almost All ◽  
Two Hybrid ◽  
Wild Type Control

In almost all symbiotic interactions between rhizobia and leguminous plants, host flavonoid–induced synthesis of Nod factors in rhizobia is required to initiate symbiotic response in plants. In this study, we found that Lotus japonicus Nod factor receptor 5 (LjNFR5) might directly regulate flavonoid biosynthesis during symbiotic interaction with rhizobia. A yeast two-hybrid analysis revealed that a dihydroflavonol-4-reductase-like protein (LjDFL1) interacts with LjNFR5. The interaction between MtDFL1 and MtNFP, two Medicago truncatula proteins with homology to LjDFL1 and LjNFR5, respectively, was also shown, suggesting that interaction between these two proteins might be conserved in different legumes. LjDFL1 was highly expressed in root hairs and epidermal cells of root tips. Lotus ljdfl1 mutants and Medicago mtdfl1 mutants produced significantly fewer infection threads (ITs) than the wild-type control plants following rhizobial treatment. Furthermore, the roots of stable transgenic L. japonicus plants overexpressing LjDFL1 formed more ITs than control roots after exposure to rhizobia. These data indicated that LjDFL1 is a positive regulator of symbiotic signaling. However, the expression of LjDFL1 was suppressed by rhizobial treatment, suggesting that a negative feedback loop might be involved in regulation of the symbiotic response in L. japonicus.

scholarly journals 3,4-dehydro-L-proline Induces Programmed Cell Death in the Roots of Brachypodium distachyon

2021 ◽  
Vol 22 (14) ◽  
pp. 7548
Author(s):  
Artur Pinski ◽  
Alexander Betekhtin ◽  
Jolanta Kwasniewska ◽  
Lukasz Chajec ◽  
Elzbieta Wolny ◽  
...  
Keyword(s):  
Cell Death ◽  
Brachypodium Distachyon ◽  
Root Hairs ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Cell Wall Proteins ◽  
Root Epidermis ◽  
Root Hair Development ◽  
Transmission Electron ◽  
Hair Development

As cell wall proteins, the hydroxyproline-rich glycoproteins (HRGPs) take part in plant growth and various developmental processes. To fulfil their functions, HRGPs, extensins (EXTs) in particular, undergo the hydroxylation of proline by the prolyl-4-hydroxylases. The activity of these enzymes can be inhibited with 3,4-dehydro-L-proline (3,4-DHP), which enables its application to reveal the functions of the HRGPs. Thus, to study the involvement of HRGPs in the development of root hairs and roots, we treated seedlings of Brachypodium distachyon with 250 µM, 500 µM, and 750 µM of 3,4-DHP. The histological observations showed that the root epidermis cells and the cortex cells beneath them ruptured. The immunostaining experiments using the JIM20 antibody, which recognizes the EXT epitopes, demonstrated the higher abundance of this epitope in the control compared to the treated samples. The transmission electron microscopy analyses revealed morphological and ultrastructural features that are typical for the vacuolar-type of cell death. Using the TUNEL test (terminal deoxynucleotidyl transferase dUTP nick end labelling), we showed an increase in the number of nuclei with damaged DNA in the roots that had been treated with 3,4-DHP compared to the control. Finally, an analysis of two metacaspases’ gene activity revealed an increase in their expression in the treated roots. Altogether, our results show that inhibiting the prolyl-4-hydroxylases with 3,4-DHP results in a vacuolar-type of cell death in roots, thereby highlighting the important role of HRGPs in root hair development and root growth.

Analysis of ENOD40 expression in alb1, a symbiotic mutant of Lotus japonicus that forms empty nodules with incompletely developed nodule vascular bundles

2000 ◽  
Vol 264 (4) ◽  
pp. 402-410 ◽  
Author(s):  
H. Imaizumi-Anraku ◽  
H. Kouchi ◽  
K. Syono ◽  
S. Akao ◽  
M. Kawaguchi
Keyword(s):  
Lotus Japonicus ◽  
Vascular Bundles

Determination of Ochratoxin A in Feed by Immunoaffinity Cleanup and Liquid Chromatography

2013 ◽  
Vol 96 (3) ◽  
pp. 599-602 ◽  
Author(s):  
Ping Ding ◽  
Ziyou Mi ◽  
Yali Hou ◽  
Yigang He ◽  
Jianhua Xie
Keyword(s):  
Liquid Chromatography ◽  
Detection Limit ◽  
Ochratoxin A ◽  
Cyanogen Bromide ◽  
Polyclonal Antibodies ◽  
Immunoaffinity Column ◽  
Low Levels ◽  
Sepharose 4B ◽  
Feed Samples

Abstract A method using LC was developed for determination of ochratoxin A (OTA) in feeds. The extracted samples were cleaned up by an immunoaffinity column prepared by covalently coupling polyclonal antibodies against OTA to cyanogen bromide-activated Sepharose 4B. The eluates were determined by LC with fluorescence detection. Recoveries of OTA from fortified samples of 1–10 μg/kg levels ranged from 84.3 to 90.0%, with CVs of 3.3–7.8%. The detection limit was 0.045 μg/kg based on an S/N of 3:1. A total of 65 feed samples were screened for OTA with the proposed method. The results showed that only nine samples were contaminated with OTAs at low levels. The presented method was successfully applied to quantify OTAs in real feed samples.

scholarly journals Molecular cloning of two human liver 3 α-hydroxysteroid/dihydrodiol dehydrogenase isoenzymes that are identical with chlordecone reductase and bile-acid binder

1994 ◽  
Vol 299 (2) ◽  
pp. 545-552 ◽  
Author(s):  
Y Deyashiki ◽  
A Ogasawara ◽  
T Nakayama ◽  
M Nakanishi ◽  
Y Miyabe ◽  
...  
Keyword(s):  
Bile Acid ◽  
Amino Acid ◽  
Human Liver ◽  
Polyclonal Antibodies ◽  
Amino Acid Sequences ◽  
Cdna Clones ◽  
Amino Acid Residues ◽  
Human Bile ◽  
Coding Regions ◽  
Computer Based

Human liver contains two dihydrodiol dehydrogenases, DD2 and DD4, associated with 3 alpha-hydroxysteroid dehydrogenase activity. We have raised polyclonal antibodies that cross-reacted with the two enzymes and isolated two 1.2 kb cDNA clones (C9 and C11) for the two enzymes from a human liver cDNA library using the antibodies. The clones of C9 and C11 contained coding sequences corresponding to 306 and 321 amino acid residues respectively, but lacked 5′-coding regions around the initiation codon. Sequence analyses of several peptides obtained by enzymic and chemical cleavages of the two purified enzymes verified that the C9 and C11 clones encoded DD2 and DD4 respectively, and further indicated that the sequence of DD2 had at least additional 16 residues upward from the N-terminal sequence deduced from the cDNA. There was 82% amino acid sequence identity between the two enzymes, indicating that the enzymes are genetic isoenzymes. A computer-based comparison of the cDNAs of the isoenzymes with the DNA sequence database revealed that the nucleotide and amino acid sequences of DD2 and DD4 are virtually identical with those of human bile-acid binder and human chlordecone reductase cDNAs respectively.

A γGT-AT1A receptor transgene protects renal cortical structure in AT1 receptor-deficient mice

2004 ◽  
Vol 18 (3) ◽  
pp. 290-298 ◽  
Author(s):  
Thu H. Le ◽  
Michael I. Oliverio ◽  
Hyung-Suk Kim ◽  
Harmony Salzler ◽  
Rajesh C. Dash ◽  
...  
Keyword(s):  
Blood Pressure ◽  
Transgenic Mice ◽  
Proximal Tubule ◽  
Physiological Role ◽  
Renal Proximal Tubule ◽  
Hypoxanthine Phosphoribosyl Transferase ◽  
Wild Type ◽  
Specific Expression ◽  
Low Levels ◽  
Deficient Mice

To understand the physiological role of angiotensin type 1 (AT1) receptors in the proximal tubule of the kidney, we generated a transgenic mouse line in which the major murine AT1 receptor isoform, AT1A, was expressed under the control of the P1 portion of the γ-glutamyl transpeptidase (γGT) promoter. In transgenic mice, this promoter has been shown to confer cell-specific expression in epithelial cells of the renal proximal tubule. To avoid random integration of multiple copies of the transgene, we used gene targeting to produce mice with a single-copy transgene insertion at the hypoxanthine phosphoribosyl transferase ( Hprt) locus on the X chromosome. The physiological effects of the γGT-AT1A transgene were examined on a wild-type background and in mice with targeted disruption of one or both of the murine AT1 receptor genes ( Agtr1a and Agtr1b). On all three backgrounds, γGT-AT1A transgenic mice were healthy and viable. On the wild-type background, the presence of the transgene did not affect development, blood pressure, or kidney structure. Despite relatively low levels of expression in the proximal tubule, the transgene blunted the increase in renin expression typically seen in AT1-deficient mice and partially rescued the kidney phenotype associated with Agtr1a−/− Agtr1b−/− mice, significantly reducing cortical cyst formation by more than threefold. However, these low levels of cell-specific expression of AT1 receptors in the renal proximal tubule did not increase the low blood pressures or abolish sodium sensitivity, which are characteristic of AT1 receptor-deficient mice. Although our studies do not clearly identify a role for AT1 receptors in the proximal tubules of the kidney in blood pressure homeostasis, they support a major role for these receptors in modulating renin expression and in maintaining structural integrity of the renal cortex.

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