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Root Nodules (root + nodule)
Selected AbstractsThe Effect of Seed Vernalization and Irradiation on Growth and Photosynthesis of Field Bean Plants (Vicia faba L. minor) and on Nitrogenase Activity of Root NodulesJOURNAL OF AGRONOMY AND CROP SCIENCE, Issue 4 2000W. Filek The effect of seed vernalization (2,3°C; 36 days) and irradiation intensity during vegetation of field bean upon growth and maximum leaf photosynthesis (Pmax) as well as specific (SNA) and total nitrogenase activity (TNA) of root nodules was investigated. The measurements were performed at the initial stage of pod development of varieties with undetermined (cv. Nadwi,a,ski) and determined (cv. Tibo) growth habit. In comparison with plants in shading, those which grew in full irradiation produced larger leaf area and more dry weight, achieving higher Pmax values. Stimulation of leaf area enlargement and dry weight increase, especially in respect to the dry weight of underground parts had a positive and significant effect upon TNA value. Accumulation of dry weight was higher in vernalized than non-vernalized plants, but only in the case of cv. Nadwi,la,ski, in full irradiation conditions. In the case of plants grown in poorer irradiation conditions TNA values were about 68 % (cv. Nadwi,la,ski) and 54 % (cv. Tibo) lower, due to significant limitation of root nodules. TNA value of vernalized cv. Nadwi,la,ski plants was 66 % higher in comparison with non-vernalized plants. However, the effect was visible only in full irradiation conditions, due to simultaneous greater intensity of root nodules production and higher SNA value. A positive effect of vernalization could also be observed in the decrease of a negative correlation between dry weight of root nodules and SNA in both varieties. Zusammenfassung Der Einfluss der Samenvernalisation (3,3°C; 36 Tage) und der Strahlungsintensität während der Vegetation von Fababohnen hinsichtlich des Wachstums und der maximalen Blattfotosynthese (Pmax) sowie der spezifischen (SNA) und Gesamtnitrogenaseaktivität (TNA) der Wurzelknöllchen wurde untersucht. Die Messungen wurden im ersten Stadium der Hülsenentwicklung an Sorten mit undeterminierten (cv. Nadwi,la,ski) und determinierten (cv. Tibo) Wachstumsstrukturen durchgeführt. Ein Vergleich von Pflanzen unter Beschattung mit Pflanzen unter voller Strahlung zeigte, dass beschattete Pflanzen eine gröiere Blattfläche und höheres Trockengewicht produzierten und damit höhere Pmax -Werte erreichten. Stimulation der Blattflächenvergröierung und der Trockenge wichtszunahme, insbesondere im Hinblick auf die Trockengewichte der unterirdischen Pflanzenteile hatte eine positive und signifikante Auswirkung auf den DNA-Wert. Die Akkumulation von Trockenmasse war in vernalisierten Pflanzen höher als in nicht vernalisierten; dies gilt nur für Sorte Nadwi,la,ski unter dem Einfluss voller Strahlung. Im Fall der Pflanzen, die unter geringer Strahlung wuchsen, waren die TNA-Werte ca. 68 % (cv.Nadwi,la,ski) und 54 % (cv. Tibo) geringer als Folge einer signifikanten Begrenzung des Wurzelknöllchenwachstums. TNA-Werte vernalisierter cv. Nadwi,la,ski-Pflanzen waren um 66 % höher im Vergleich mit nicht vernalisierten Pflanzen. Allerdings zeigte sich diese Wirkung nur unter voller Strahlung als Folge einer vergleichsweise gröieren Intensität der Wurzelknöllchenproduktion und höherer SNA-Werte. Der positive Effekt der Vernalisation konnte ebenfalls beobachtet werden bei einer Abnahme der negativen Korrelation zwischen dem Trockengewicht der Wurzelknöllchen und SNA in beiden Sorten. [source] Effect of Salt Stress on Carbon Metabolism and Bacteroid Respiration in Root Nodules of Common Bean (Phaseolus vulgaris L.)PLANT BIOLOGY, Issue 4 2000A. Ferri Abstract: In the present work, we examined the effect of salinity on growth, N fixation and carbon metabolism in the nodule cytosol and bacteroids of Phaseolus vulgaris, and measured the O2 consumption by bacteroids incubated with or without the addition of exogenous respiratory substrates. The aim was to ascertain whether the compounds that accumulate under salt stress can increase bacteroid respiration and whether this capacity changes in response to salinity in root nodules of Phaseolus vulgaris. The plants were grown in a controlled environment chamber, and 50, 100 mM or no NaCl (control) was added to the nutrient solution. Two harvests were made, at the vegetative growth period and at the beginning of the reproductive period. The enzyme activities in the nodule cytosol were reduced by the salt treatments, while in the bacteroid cytosol the enzyme activities increased at high salt concentrations at the first harvest and for ADH in all treatments. The data presented here confirm that succinate and malate are the preferred substrates for bacteroid respiration in common bean, but these bacteroids may also utilize glucose, either in control or under saline conditions. The addition of proline or lactate to the incubation medium significantly raised oxygen consumption in the bacteroids isolated from plants treated with salt. [source] Direct amplification of rhizobial nodC sequences from soil total DNA and comparison to nodC diversity of root nodule isolatesFEMS MICROBIOLOGY ECOLOGY, Issue 1 2005Sarita Sarita Abstract A group-specific primer set was developed using nodC as a target gene for the amplification of rhizobial sequence diversity from nodule isolates and total soil DNA preparations. The primer set was tested on 209 nodule isolates, recovered from six different trap plant species which were grown in two soil samples collected from a chickpea and a wheat field site in India. We also amplified and cloned PCR products from total DNA isolated from the same soil samples. The total diversity within the resulting clone libraries (, 218 clones) was higher than that recovered from trap plants, but differed depending on the PCR protocols and primers used. However, some plant-selected genotypes could not be obtained using the community approach, probably due to variable detection limits and limited clone library sizes. [source] A nodule-specific plant cysteine proteinase, AsNODF32, is involved in nodule senescence and nitrogen fixation activity of the green manure legume Astragalus sinicusNEW PHYTOLOGIST, Issue 1 2008Yixing Li Summary ,,Asnodf32, encoding a nodule-specific cysteine proteinase in Astragalus sinicus, is probably involved in nodule senescence. To obtain direct evidence of its role in nodule senescence, Agrobacterium rhizogenes -mediated RNA interference was applied to A. sinicus hairy roots. ,,Real-time qRT-PCR was used to estimate the efficiency of suppression. The senescent phenotype of transgenic nodules was examined with paraffin-embedded slides, TUNEL (TdT-mediated dUTP nick-end labeling) assay, and transmission electron microscopy, and the bacteroid nitrogen fixation activity was also measured. ,,It was found that silencing of Asnodf32 delayed root nodule and bacteroid senescence. The period of bacteroid active nitrogen fixation was significantly extended. Interestingly, nodules enlarged in length were also observed on Asnodf32- silenced hairy roots. ,,The results reported here indicate that Asnodf32 plays an important role in the regulation of root nodule senescence. [source] Influence of an extract from kudzu symbiosomes containing leghemoglobin on in vitro cutaneous procollagen productionINTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 3 2010J. V. Gruber J. Cosmet. Sci., 60, 475,484 (September/October 2009) Synopsis Cytoglobin is a hexacoordinateglobin protein that was recently discovered in mammals. Interestingly, of the four human globin proteins that are now known, hemoglobin, myoglobin, neuroglobin and cytoglobin, the latter appears to have the closest resemblance to strikingly similar proteins expressed in plants. In legumes, these proteins accumulate in symbiosomes (root nodules) of various legumes and are called leghemoglobin. The paper will discuss the ability of an aqueous extract from Pueraria lobata (kudzu) symbiosomes that contains leghemoglobin to stimulate procollagen production in human dermal fibroblasts. This effect may be partly due to the possibility that leghemoglobin may mimic the function of cytoglobin by shuttling oxygen to prolyl-4-hydroxylase, the enzyme responsible for oxidizing proline residues in procollagen bundles. This hypothesis is supported by DNA microarray sequencing data that demonstrate that treatment of normal human dermal fibroblasts (NHDF) with highly purified cytoglobin or leghemoglobin upregulates a number of key collagen-related genes including COL1A1 and COL1A2. [source] The Effect of Seed Vernalization and Irradiation on Growth and Photosynthesis of Field Bean Plants (Vicia faba L. minor) and on Nitrogenase Activity of Root NodulesJOURNAL OF AGRONOMY AND CROP SCIENCE, Issue 4 2000W. Filek The effect of seed vernalization (2,3°C; 36 days) and irradiation intensity during vegetation of field bean upon growth and maximum leaf photosynthesis (Pmax) as well as specific (SNA) and total nitrogenase activity (TNA) of root nodules was investigated. The measurements were performed at the initial stage of pod development of varieties with undetermined (cv. Nadwi,a,ski) and determined (cv. Tibo) growth habit. In comparison with plants in shading, those which grew in full irradiation produced larger leaf area and more dry weight, achieving higher Pmax values. Stimulation of leaf area enlargement and dry weight increase, especially in respect to the dry weight of underground parts had a positive and significant effect upon TNA value. Accumulation of dry weight was higher in vernalized than non-vernalized plants, but only in the case of cv. Nadwi,la,ski, in full irradiation conditions. In the case of plants grown in poorer irradiation conditions TNA values were about 68 % (cv. Nadwi,la,ski) and 54 % (cv. Tibo) lower, due to significant limitation of root nodules. TNA value of vernalized cv. Nadwi,la,ski plants was 66 % higher in comparison with non-vernalized plants. However, the effect was visible only in full irradiation conditions, due to simultaneous greater intensity of root nodules production and higher SNA value. A positive effect of vernalization could also be observed in the decrease of a negative correlation between dry weight of root nodules and SNA in both varieties. Zusammenfassung Der Einfluss der Samenvernalisation (3,3°C; 36 Tage) und der Strahlungsintensität während der Vegetation von Fababohnen hinsichtlich des Wachstums und der maximalen Blattfotosynthese (Pmax) sowie der spezifischen (SNA) und Gesamtnitrogenaseaktivität (TNA) der Wurzelknöllchen wurde untersucht. Die Messungen wurden im ersten Stadium der Hülsenentwicklung an Sorten mit undeterminierten (cv. Nadwi,la,ski) und determinierten (cv. Tibo) Wachstumsstrukturen durchgeführt. Ein Vergleich von Pflanzen unter Beschattung mit Pflanzen unter voller Strahlung zeigte, dass beschattete Pflanzen eine gröiere Blattfläche und höheres Trockengewicht produzierten und damit höhere Pmax -Werte erreichten. Stimulation der Blattflächenvergröierung und der Trockenge wichtszunahme, insbesondere im Hinblick auf die Trockengewichte der unterirdischen Pflanzenteile hatte eine positive und signifikante Auswirkung auf den DNA-Wert. Die Akkumulation von Trockenmasse war in vernalisierten Pflanzen höher als in nicht vernalisierten; dies gilt nur für Sorte Nadwi,la,ski unter dem Einfluss voller Strahlung. Im Fall der Pflanzen, die unter geringer Strahlung wuchsen, waren die TNA-Werte ca. 68 % (cv.Nadwi,la,ski) und 54 % (cv. Tibo) geringer als Folge einer signifikanten Begrenzung des Wurzelknöllchenwachstums. TNA-Werte vernalisierter cv. Nadwi,la,ski-Pflanzen waren um 66 % höher im Vergleich mit nicht vernalisierten Pflanzen. Allerdings zeigte sich diese Wirkung nur unter voller Strahlung als Folge einer vergleichsweise gröieren Intensität der Wurzelknöllchenproduktion und höherer SNA-Werte. Der positive Effekt der Vernalisation konnte ebenfalls beobachtet werden bei einer Abnahme der negativen Korrelation zwischen dem Trockengewicht der Wurzelknöllchen und SNA in beiden Sorten. [source] Phenotypic characteristics of rhizobia isolates nodulating Acacia species in the arid and Saharan regions of MoroccoLETTERS IN APPLIED MICROBIOLOGY, Issue 5 2000K. Zerhari The phenotypic characteristics of 48 isolates obtained from root nodules of four Acacia species (Acacia cyanophylla, A. gummifera, A. horrida and A. raddiana) growing in soils collected from the arid and Saharan regions of Morocco were studied. The rhizobia were very diverse with respect to their cross-nodulation patterns, as well as their physiological and biochemical properties. Dendrograms obtained through computer numerical analysis of 52 phenotypic characteristics showed that isolates could fit into four clusters below the boundary level of 0·85 average distance and that they were very distinct from the reference strains. Some interesting isolates for inoculation trials have been identified. They were able to grow at pH ranging from 4 to 9, tolerate a high salt concentration (3% NaCl) and grew at a maximum temperature between 35 and 40 °C. [source] Importance of glutathione in the nodulation process of peanut (Arachis hypogaea)PHYSIOLOGIA PLANTARUM, Issue 2 2008Eliana Bianucci GSH appears to be essential for proper development of the root nodules during the symbiotic association of legume,rhizobia in which the entry of rhizobia involves the formation of infection threads. In the particular case of peanut,rhizobia symbiosis, the entry of rhizobia occurs by the mechanism of infection called ,crack entry', i.e. entry at the point of emergence of lateral roots. We have previously shown the role of GSH content of Bradyrhizobium sp. SEMIA 6144 during the symbiotic association with peanut using a GSH-deficient mutant obtained by disruption of the gshA gene, encoding ,-glutamylcysteine synthetase (,-GCS), which was able to induce nodules in peanut roots without alterations in the symbiotic phenotype. To investigate the role of the peanut GSH content in the symbiosis, the compound l -buthionine-sulfoximine (BSO), a specific inhibitor of ,-GCS in plants, was used. There were no differences in the plant growth and the typical anatomic structure of the peanut roots when the plants grew in the Fahraeus medium either in presence or in absence of 0.1 mM BSO. However, the GSH content was reduced by 51% after treatment with BSO. The BSO-treated plants inoculated with wild-type or mutant strains of Bradyrhizobium sp. showed a significant reduction in the number and dry weight of nodules, suggesting that GSH content could play an important role in the nodulation process of root peanut with Bradyrhizobium sp. [source] The cell-cycle promoter cdc2aAt from Arabidopsis thaliana is induced in the lateral roots of the actinorhizal tree Allocasuarina verticillata during the early stages of the symbiotic interaction with FrankiaPHYSIOLOGIA PLANTARUM, Issue 3 2007Mame Ourèye Sy The symbiosis between the actinorhizal tree Allocasuarina verticillata and the actinomycete Frankia leads to the formation of root nodules inside which bacteria fix atmospheric nitrogen. Actinorhizal nodule organogenesis starts with the induction of cell divisions in the root cortex and in the pericycle cells opposite protoxylem poles near Frankia -infected root hairs. To study the ability of Frankia to induce progression through the cell cycle, we monitored the expression of the ,-glucuronidase (gus) gene driven by the promoter from cdc2aAt, an Arabidopsis cyclin-dependent kinase gene that displays competence for cell division, during plant growth and nodule ontogenesis. In non-symbiotic tissues, the gus gene was mainly expressed in primary and secondary meristems of roots and shoots. Auxins and cytokinins were found to induce reporter gene activity in the root system of whole plants, showing that the promoter cdc2aAt displayed the same regulation by hormones in Allocasuarina as that reported in Arabidopsis. In transgenic nodules, gus expression was found to be restricted to the phellogen. During the early stages of the interaction between Frankia and the plant root system, cdc2aAt was strongly induced in the lateral roots surrounded by hyphae of the actinomycete. Histochemical analysis of ,-glucuronidase activity revealed that cells from the pericycle opposite protoxylem poles were very deeply stained. These data indicate that upon Frankia infection, cells from the lateral roots, and notably pericycle cells that can give rise to a nodule or a root primordium, prepare to re-enter the cell cycle. [source] Trehalose metabolism in root nodules of the model legume Lotus japonicus in response to salt stressPHYSIOLOGIA PLANTARUM, Issue 4 2006Miguel López The effect of NaCl stress (50 mM) and validamycin A treatment (30 ,M) on growth and nitrogen fixation of Lotus japonicus was investigated in plants cultured under symbiotic and hydroponics conditions for teen weeks (flowering stage). Validamycin A was used as a potent trehalase inhibitor, and was able to produce a five-fold increase in the level of trehalose during salt treatment, concomitant with an enhance in biomass (20%) in salinized plants. Alterations of nodule metabolism related to some carbohydrates and some enzyme activities were also examined. The shoot and total plant dry weight were severely affected by saline conditions decreasing by 40% and only 15,20% in plant treated without or with validamycin A, respectively. Nitrogenase activity (E.C. 1.7.9.92) was inhibited almost 40% by salt stress and no effect of validamycin was observed. Based on these results, L. japonicus might be considered as a salt-sensitive legume. In addition, the saline conditions also inhibited the enzyme activities of sucrose synthase (E.C. 2.4.1.13), alkaline invertase (E.C. 3.2.1.26) and trehalose-phosphate synthetase (E.C. 2.4.1.15). The validamycin A treatment mainly decreased enzyme activities: sucrose synthase, trehalose-phosphate phosphatase (E.C. 3.1.3.12) and trehalase (E.C. 3.2.1.28). On the other hand, a high concentration of the carbohydrates, starch, sucrose and glucose, seems not to be the mechanism induced in L. japonicus to protect nodules exposed to NaCl because all these sugars decreased in such conditions. Results of the present study support the possible role of trehalose as an osmoprotectant under salt stress. [source] Rhizobium colonization induced changes in membrane-bound and soluble hydroxyproline-rich glycoprotein composition in peaPHYSIOLOGIA PLANTARUM, Issue 4 2002Pål Axel Olsson Abundance and distribution of plant cell surface proteins of the hydroxyproline-rich glycoprotein (HRGP) class were studied in the pea- Rhizobium symbiosis using immunoblot analysis. The MAC 265-epitope was especially abundant in pea root nodules containing nitrogen-fixing Rhizobium bacteria. A 180-kDa MAC 265-HRGP dominated in pea shoot plasma membranes, while almost no MAC 265-HRGP was detected in root plasma membranes. We show here that a major difference between the plant-derived peribacteroid membrane of the symbiosomes and the root plasma membrane was the presence of a 100-kDa MAC 265-HRGP in the former. Arabinogalactan proteins (AGPs), as recognized by the monoclonal antibodies MAC 207 and JIM 8, were not detected in the peribacteroid membrane, while two isoforms (100 and 220 kDa) were detected in shoot and root plasma membranes. Specific MAC 265-HRGP isoforms were found in the peribacteroid space fraction of the symbiosomes and thus as soluble proteins in the interface between the symbionts. The abundance of the MAC 265-epitope was much reduced in non-nitrogen-fixing nodules when this phenotype resulted from a dicarboxylate transport mutation in Rhizobium. There was no reduction in the abundance of the MAC 265-epitope in non-fixing phenotypes resulting from a mutation in the plant. The results suggest that bacterial signals related to the bacterial ability to fix nitrogen, might be responsible for the regulation of HRGP expression in root nodules. [source] Reactive oxygen species and antioxidants in legume nodulesPHYSIOLOGIA PLANTARUM, Issue 4 2000Manuel Becana Reactive oxygen species are a ubiquitous danger for aerobic organisms. This risk is especially elevated in legume root nodules due to the strongly reducing conditions, the high rates of respiration, the tendency of leghemoglobin to autoxidize, the abundance of nonprotein Fe and the presence of several redox proteins that leak electrons to O2. Consequently, nodules are particularly rich in both quantity and diversity of antioxidant defenses. These include enzymes such as superoxide dismutase (EC 1.15.1.1) and ascorbate peroxidase (EC 1.11.1.11) and metabolites such as ascorbate and thiol tripeptides. Nodule antioxidants have been the subject of intensive molecular, biochemical and functional studies that are reviewed here. The emerging theme is that antioxidants are especially critical for the protection and optimal functioning of N2 fixation. We hypothesize that this protection occurs at least at two levels: the O2 diffusion barrier in the nodule parenchyma (inner cortex) and the infected cells in the central zone. [source] Effect of Salt Stress on Carbon Metabolism and Bacteroid Respiration in Root Nodules of Common Bean (Phaseolus vulgaris L.)PLANT BIOLOGY, Issue 4 2000A. Ferri Abstract: In the present work, we examined the effect of salinity on growth, N fixation and carbon metabolism in the nodule cytosol and bacteroids of Phaseolus vulgaris, and measured the O2 consumption by bacteroids incubated with or without the addition of exogenous respiratory substrates. The aim was to ascertain whether the compounds that accumulate under salt stress can increase bacteroid respiration and whether this capacity changes in response to salinity in root nodules of Phaseolus vulgaris. The plants were grown in a controlled environment chamber, and 50, 100 mM or no NaCl (control) was added to the nutrient solution. Two harvests were made, at the vegetative growth period and at the beginning of the reproductive period. The enzyme activities in the nodule cytosol were reduced by the salt treatments, while in the bacteroid cytosol the enzyme activities increased at high salt concentrations at the first harvest and for ADH in all treatments. The data presented here confirm that succinate and malate are the preferred substrates for bacteroid respiration in common bean, but these bacteroids may also utilize glucose, either in control or under saline conditions. The addition of proline or lactate to the incubation medium significantly raised oxygen consumption in the bacteroids isolated from plants treated with salt. [source] A novel procedure for gentle isolation and separation of intact infected and uninfected protoplasts from the central tissue of Vicia faba L. root nodulesPLANT CELL & ENVIRONMENT, Issue 7 2003E. PEITER ABSTRACT The central tissue of Vicia faba L. root nodules is composed of cells infected with Rhizobium bacteroids and uninfected cells. For the study of various processes, such as plasma membrane transport, it is essential to separate both cell types. Initial attempts to isolate protoplasts according to protocols described in the literature resulted in non-spherical and osmotically inactive material, which is in agreement with previous descriptions. In the study reported herein, it was shown that the plasma membrane of non-spherical infected protoplasts is not intact. A new isolation and separation protocol was developed, based on dissection of the nodule prior to cell wall digestion, non-shaking digestion in hypertonic medium, and a combined procedure for release of protoplasts into slightly hypotonic medium and separation of protoplast fractions by isopycnic density gradient centrifugation. Infected and uninfected protoplasts that were isolated according to this protocol were spherical, osmotically active and excluded propidium iodide, confirming the intactness of their plasma membrane. The common fluorescein diacetate test was shown to be artefactual in infected cells, since viable bacteroids also stain in defective cells. Light and electron microscopic examination of infected protoplasts showed that protoplasts still contained starch after isolation and bacteroids in intact protoplasts had unusually high amounts of polyhydroxybutyrate. The vacuoles of infected protoplasts contained protein and membrane-enclosed structures, and were of non-acid pH; traits that are typical of protein storage vacuoles. [source] Analysis of B function in legumes: PISTILLATA proteins do not require the PI motif for floral organ development in Medicago truncatulaTHE PLANT JOURNAL, Issue 1 2009Reyes Benlloch Summary The B-class gene PISTILLATA (PI) codes for a MADS-box transcription factor required for floral organ identity in angiosperms. Unlike Arabidopsis, it has been suggested that legume PI genes contribute to a variety of processes, such as the development of floral organs, floral common petal,stamen primordia, complex leaves and N-fixing root nodules. Another interesting feature of legume PI homologues is that some of them lack the highly conserved C-terminal PI motif suggested to be crucial for function. Therefore, legume PI genes are useful for addressing controversial questions on the evolution of B-class gene function, including how they may have diverged in both function and structure to affect different developmental processes. However, functional analysis of legume PI genes has been hampered because no mutation in any B-class gene has been identified in legumes. Here we fill this gap by studying the PI function in the model legume species Medicago truncatula using mutant and RNAi approaches. Like other legume species, M. truncatula has two PI homologues. The expression of the two genes, MtPI and MtNGL9, has strongly diverged, suggesting differences in function. Our analyses show that these genes are required for petal and stamen identity, where MtPI appears to play a predominant role. However, they appear not to be required for development of the nodule, the common primordia or the complex leaf. Moreover, both M. truncatula PI homologues lack the PI motif, which indicates that the C-terminal motif is not essential for PI activity. [source] A CDPK isoform participates in the regulation of nodule number in Medicago truncatulaTHE PLANT JOURNAL, Issue 6 2006Pablo R. Gargantini Summary Medicago spp. are able to develop root nodules via symbiotic interaction with Sinorhizobium meliloti. Calcium-dependent protein kinases (CDPKs) are involved in various signalling pathways in plants, and we found that expression of MtCPK3, a CDPK isoform present in roots of the model legume Medicago truncatula, is regulated during the nodulation process. Early inductions were detected 15 min and 3,4 days post-inoculation (dpi). The very early induction of CPK3 messengers was also present in inoculated M. truncatuladmi mutants and in wild-type roots subjected to salt stress, indicating that this rapid response is probably stress-related. In contrast, the later response was concomitant with cortical cell division and the formation of nodule primordia, and was not observed in wild-type roots inoculated with nod,, strains. This late induction correlated with a change in the subcellular distribution of CDPK activities. Accordingly, an anti- MtCPK3 antibody detected two bands in soluble root extracts and one in the particulate fraction. CPK3::GFP fusions are targeted to the plasma membrane in epidermal onion cells, a localization that depends on myristoylation and palmitoylation sites of the protein, suggesting a dual subcellular localization. MtCPK3 mRNA and protein were also up-regulated by cytokinin treatment, a hormone linked to the regulation of cortical cell division and other nodulation-related responses. An RNAi-CDPK construction was used to silence CPK3 in Agrobacterium rhizogenes -transformed roots. Although no major phenotype was detected in these roots, when infected with rhizobia, the total number of nodules was, on average, twofold higher than in controls. This correlates with the lack of MtCPK3 induction in the inoculated super-nodulator sunn mutant. Our results suggest that CPK3 participates in the regulation of the symbiotic interaction. [source] Review: Correlations between oxygen affinity and sequence classifications of plant hemoglobins,BIOPOLYMERS, Issue 12 2009Benoit J. Smagghe Abstract Plants express three phylogenetic classes of hemoglobins (Hb) based on sequence analyses. Class 1 and 2 Hbs are full-length globins with the classical eight helix Mb-like fold, whereas Class 3 plant Hbs resemble the truncated globins found in bacteria. With the exception of the specialized leghemoglobins, the physiological functions of these plant hemoglobins remain unknown. We have reviewed and, in some cases, measured new oxygen binding properties of a large number of Class 1 and 2 plant nonsymbiotic Hbs and leghemoglobins. We found that sequence classification correlates with distinct extents of hexacoordination with the distal histidine and markedly different overall oxygen affinities and association and dissociation rate constants. These results suggest strong selective pressure for the evolution of distinct physiological functions. The leghemoglobins evolved from the Class 2 globins and show no hexacoordination, very high rates of O2 binding (,250 ,M,1 s,1), moderately high rates of O2 dissociation (,5,15 s,1), and high oxygen affinity (Kd or P50 , 50 nM). These properties both facilitate O2 diffusion to respiring N2 fixing bacteria and reduce O2 tension in the root nodules of legumes. The Class 1 plant Hbs show weak hexacoordination (KHisE7 , 2), moderate rates of O2 binding (,25 ,M,1 s,1), very small rates of O2 dissociation (,0.16 s,1), and remarkably high O2 affinities (P50 , 2 nM), suggesting a function involving O2 and nitric oxide (NO) scavenging. The Class 2 Hbs exhibit strong hexacoordination (KHisE7 , 100), low rates of O2 binding (,1 ,M,1 s,1), moderately low O2 dissociation rate constants (,1 s,1), and moderate, Mb-like O2 affinities (P50 , 340 nM), perhaps suggesting a sensing role for sustained low, micromolar levels of oxygen. © 2009 Wiley Periodicals, Inc. Biopolymers 91: 1083,1096, 2009. This article was originally published online as an accepted preprint. The "Published Online" date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com [source] |