Common Bean Roots Research Articles

Two effects of combined nitrogen on the adhesion of Rhizobium etli to bean roots

Combined forms of nitrogen negatively influence rhizobia-legume symbiosis. The effects of combined nitrogen are known for nodulation and dinitrogen (N2) fixation, but little is known about the effect on preinfection events. Here, we studied the effects of combined nitrogen on the adhesion of Rhizobium etli to common bean (Phaseolus vulgaris L.) roots. When potassium nitrate (KNO3) or sodium glutamate was added to an incubation mixture of rhizobia and plants that were previously grown in nitrogen-free solution, rhizobial adhesion to roots was stimulated. However, the rhizobial adhesion to bean roots that were previously grown with 10 mM KNO3 was reduced by half. A fraction of the bean root exudates, which is thermolabile and has molecular mass larger than 12 kDa stimulated rhizobial adhesion, but this stimulatory activity was lost in root exudates obtained with 10 mM KNO3. Thus, the inhibition of symbiosis in response to combined nitrogen may be controlled by the plant at the preinfection stage as well.

Genetic diversity of rhizobia nodulating common bean (Phaseolus vulgaris L.) in the Central Black Sea Region of Turkey

We have analyzed 30 rhizobial isolates obtained from common bean (Phaseolus vulgaris L.) root nodules grown in the Middle Blacksea Region of Turkey, using ARDRA and nucleotide sequence data. ARDRA analysis with enzymes CfoI, HinfI, NdeII, MspI and PstI revealed three patterns. Based on sequence data from 16S rDNA, the patterns were identified as, Rhizobium leguminosarum bv. phaseoli (n = 16), R. etli bv. phaseoli (n = 8) and R. phaseoli (n = 6). On the other hand, nucleotide sequence phylogenies of housekeeping genes (recA, atpD and glnII) selected to confirm the 16S rDNA phylogeny revealed different evolutionary relationships. These results suggested the possibility of lateral transfers of these genes amongst different rhizobial species (including R. leguminosarum, R. etli and R. phaseoli) sharing the same ecological niche (nodulating P. vulgaris) which also indicates that there may be no true genetic barier among these species. Phylogenetic analysis based on DNA sequence data from the nodA and nifH genes showed that all rhizobial species obtained in this study were carrying nodA and nifH haplotypes which were the same or similar to those of CFN42 (R. etli type strain), suggesting a further support for the lateral transfer of CFN42 Sym plasmid, p42, amongst Turkish common bean nodulating rhizobial isolates.

Analysis of the Proteome of Common Bean (Phaseolus vulgaris L.) Roots after Inoculation with Rhizobium etli

Proteomics techniques were used to identify the underlying mechanism of the early stage of symbiosis between the common bean (Phaseolus vulgaris L.) and bacteria. Proteins from roots of common beans inoculated with bacteria were separated using two-dimensional polyacrylamide gel electrophoresis and identified using mass spectrometry. From 483 protein spots, 29 plant and 3 bacterial proteins involved in the early stage of symbiosis were identified. Of the 29 plant proteins, the expression of 19 was upregulated and the expression of 10 was downregulated. Upregulated proteins included those involved in protein destination/storage, energy production, and protein synthesis; whereas the downregulated proteins included those involved in metabolism. Many upregulated proteins involved in protein destination/storage were chaperonins and proteasome subunits. These results suggest that defense mechanisms associated with induction of chaperonins and protein degradation regulated by proteasomes occur during the early stage of symbiosis between the common bean and bacteria.

Parallel Sequencing of Expressed Sequence Tags from Two Complementary DNA Libraries for High and Low Phosphorus Adaptation in Common Beans

Expressed sequence tags (ESTs) have proven useful for gene discovery in many crops. In this work, our objective was to construct complementary DNA (cDNA) libraries from root tissues of common beans (Phaseolus vulgaris L.) grown under low and high P hydroponic conditions and to conduct EST sequencing and comparative analyses of the libraries. Expressed sequence tag analysis of 3648 clones identifi ed 2372 unigenes, of which 1591 were annotated as known genes while a total of 465 unigenes were not associated with any known gene. Unigenes with hits were categorized according to biological processes, molecular function, and cellular compartmentalization. Given the young tissue used to make the root libraries, genes for catalytic activity and binding were highly expressed. Comparisons with previous root EST sequencing and between the two libraries made here resulted in a set of genes to study further for differential gene expression and adaptation to low P, such as a 14 kDa praline-rich protein, a metallopeptidase, tonoplast intrinsic protein, adenosine triphosphate (ATP) citrate synthase, and cell proliferation genes expressed in the low P treated plants. Given that common beans are often grown on acid soils of the tropics and subtropics that are usually low in P these genes and the two parallel libraries will be useful for selection for better uptake of this essential macronutrient. The importance of EST generation for common bean root tissues under low P and other abiotic soil stresses is also discussed. E XPRESSED SEQUENCE TAGS (ESTs) are the sequences of complementary DNA (cDNA) clones that represent transcribed genes (Hatey et al., 1998). Th ese sequences are derived from cDNA libraries and collections of cDNA clones created from cloning messenger RNA (mRNA) through reverse transcription and ligation to a vector. Expressed sequence tags are considered to be part of the transcriptome and to represent to a greater or lesser degree the genes expressed in the genotypes and tissues at the time point at which the mRNA was extracted. Th erefore, a large eff ort has gone into sequencing cDNA libraries from many diff erent types of tissues under many diff erent treatments in many diff erent

Nodulation and nitrogen fixation in interspecies grafts of soybean and common bean is controlled by isoflavonoid signal molecules translocated from shoot

  Identification of common signals of nodulation control among legume species will facilitate progress in enhancing symbiotic nitrogen fixation of legumes in sustainable agriculture system. Grafting experiments between soybean and common bean were carried out to evaluate whether a common shoot signals control the expression of hypernodulation among the tow species. Grafting of a hypernodulating soybean mutant NOD1-3 shoots to three cultivars of normally nodulating common bean roots resulted in hypernodulation on roots of three tested cultivars of common bean. The shoot control of hypernodulation may be causally related to differential root isoflavonoid levels, which are also controlled by shoot factors. Isoflavonoid analysis from root extracts of grafted plants showed that NOD1-3 shoots had markedly higher root isoflavonoid concentrations in roots of both NOD1-3 and common bean cv. Adzuki compared with self-grafts of common bean Adzuki. Exogenous application of daidzein, genistein, coumestrol, glycitein and in combination at concentration of 10 µmol to the nutrient solution significantly increased the nodule numbers of common bean cv. Adzuki. Therefore, the control of hypernodulation expression by isoflavonoid signal molecules translocated from shoot is common among legume species.  

Nodulin 41, a novel late nodulin of common bean with peptidase activity.

BackgroundThe legume-rhizobium symbiosis requires the formation of root nodules, specialized organs where the nitrogen fixation process takes place. Nodule development is accompanied by the induction of specific plant genes, referred to as nodulin genes. Important roles in processes such as morphogenesis and metabolism have been assigned to nodulins during the legume-rhizobium symbiosis.ResultsHere we report the purification and biochemical characterization of a novel nodulin from common bean (Phaseolus vulgaris L.) root nodules. This protein, called nodulin 41 (PvNod41) was purified through affinity chromatography and was partially sequenced. A genomic clone was then isolated via PCR amplification. PvNod41 is an atypical aspartyl peptidase of the A1B subfamily with an optimal hydrolytic activity at pH 4.5. We demonstrate that PvNod41 has limited peptidase activity against casein and is partially inhibited by pepstatin A. A PvNod41-specific antiserum was used to assess the expression pattern of this protein in different plant organs and throughout root nodule development, revealing that PvNod41 is found only in bean root nodules and is confined to uninfected cells.ConclusionsTo date, only a small number of atypical aspartyl peptidases have been characterized in plants. Their particular spatial and temporal expression patterns along with their unique enzymatic properties imply a high degree of functional specialization. Indeed, PvNod41 is closely related to CDR1, an Arabidopsis thaliana extracellular aspartyl protease involved in defense against bacterial pathogens. PvNod41's biochemical properties and specific cell-type localization, in uninfected cells of the common bean root nodule, strongly suggest that this aspartyl peptidase has a key role in plant defense during the symbiotic interaction.

Spatio-temporal analysis of development of basal roots of common bean (Phaseolus vulgaris L.)

Temporal development of roots is key to the understanding of root system architecture of plants which influences nutrient uptake, anchorage and plant competition. Using time lapse imaging we analyzed developmental patterns of length, growth angle, depth and curvature of Phaseolus basal roots from emergence till 48 h in two genotypes, B98311 and TLP19 with contrasting growth angles. In both genotypes all basal roots appeared almost simultaneously, but their growth rates varied which accounted for differences in root length. The growth angles of the basal roots fluctuated rapidly during initial development due to oscillatory root growth causing local bends. Beyond 24 h, as the root curvature stabilized, so did the growth angle. Therefore growth angle of basal roots is not a very reliable quantity for characterizing root architecture, especially during early seedling development. Comparatively, tip depth is a more robust measure of vertical distribution of the basal roots even during early seedling development.

Induction of Resistance in Common Bean by Rhizobium leguminosarum bv. phaseoli and Decrease of Common Bacterial Blight

Normal 0 14 false false false IT ZH-TW X-NONE MicrosoftInternetExplorer4 Rhizobium leguminosarum bv. phaseoli was evaluated for its capacity to trigger resistance to common bacterial blight (CBB) of common bean caused by Xanthomonas axonopodis pv. phaseoli (Xap), under greenhouse and field conditions. A common bean cultivar and three lines including two tolerant lines (Ks51103 and BF13607) and the susceptible cultivar Khomein and line Ks21479, were used. R. leguminosarum bv. phaseoli , was applied as a seed treatment and its effect on disease severity (DS) was compared with untreated (control) plants and with urea fertilizer treated plants in both the greenhouse and the feld. R. leguminosarum bv. phaseoli in common bean roots tended to reduce CBB severity and to improve plant growth, particularly the 100-seed weight, in the field. The greatest decrease in CBB in the greenhouse occurred 15 days after inoculation (DAI) of Xap in the cultivar Khomein and line Ks21479, and 30 DAI in lines Ks51103 and BF13607. In the field, however, the greatest decrease occurred 35 DAI in the cultivar and all lines. Furthermore, in the field the greatest improvement in 100-seed weight occurred in the cultivar Khomein treated with R. leguminosarum bv. phaseoli . The potential of using R. leguminosarum bv. phaseoli in the management of CBB and the possible mechanisms that induce resistance in this symbiotic system are discussed.

Anatomical root variations in response to water deficit: wild and domesticated common bean (Phaseolus vulgaris L).

Root anatomical responses to water deficit are diverse and regulation of water uptake strongly depends on plant anatomy. The ancestors of common bean (Phaseolus vulgaris L.) cultivars are the wild common beans. Because wild beans adapt and survive well in the natural environment, it is hypothesized that wild common bean roots are less affected than those of domesticated beans at low substrate water potential (ψW). A wild common bean accession from Chihuahua Mexico and cv. Bayomex were studied. Seedlings with a mean root length between 3 and 4 cm were maintained for 24 h in vermiculite at ψW of -0.03 (well hydrated), -0.65, -1.48 and -2.35 MPa (partially dry). Ten anatomical characteristics of differentiation and cell division in root regions were evaluated. Thickness of epidermis and protoderm diminished similarly in wild and domesticated beans growing at low substrate ψW (between -0.65 and -2.35 MPa). At the same time, parenchymatic cell area diminished by 71 % in the domesticated variety, but by only 32 % in the wild bean at -2.35 MPa. The number of cells in the cortex and the thickness of the xylem wall increased in both wild and domesticated beans at low substrate ψW; nevertheless, the effect was significantly lower in the wild bean. The number of xylem vessels increased in the cultivar (up to 40 %) while in the wild bean it decreased (up to 33 %). The diameter of xylem vessels and transverse root area diminished (15 and 57 %, respectively) in the cultivar, but in the wild common bean were not affected. Anatomical root characteristics and their modifications in both differentiation and cell division in root regions demonstrated that the wild bean reacted quite differently to substrate ψW than the domesticated common bean.

Evaluation of 2,4-D and Dicamba genotoxicity in bean seedlings using comet and RAPD assays

The present study was undertaken to evaluate genotoxic potential of two auxinic herbicides [2,4-dicholorophenoxy acetic acid (2,4-D) and 3,6-dichloro-2-methoxybenzoic acid (Dicamba)] in the roots of common bean ( Phaseolus vulgaris L.) seedlings. Two-day-old etiolated seedlings were treated with 10 ppm methyl methanesulfonate (MMS, positive control) or 0.1, 0.2, or 0.3 ppm of either 2,4-D or Dicamba. At the end of a 96 h growth period, root growth, total soluble protein content, DNA damage in individual cells (comet assay scores) and randomly amplified polymorphic DNA (RAPD) profiles were used as endpoints of genotoxicity. 2,4-D and Dicamba were clearly dose-dependent root growth inhibitors. Total soluble protein content was significantly decreased in the positive control and at high concentrations (0.2 and 0.3 ppm) of Dicamba. Soluble protein content increased significantly only at 0.3 ppm 2,4-D ( P<0.05). In the comet assay, DNA fragmentation increased in a dose-dependent manner. The diagnostic and phenetic analyzes of appeared and/or disappeared RAPD bands indicated that dose-dependent DNA polymorphism was induced by both herbicides. Genomic template stability was significantly affected at all 2,4-D and Dicamba doses tested. Overall 2,4-D and Dicamba have similar effects on DNA damage detected by comet and RAPD assays.

Transcript profiling of common bean (Phaseolus vulgaris L.) using the GeneChip(R) Soybean Genome Array: optimizing analysis by masking biased probes

BackgroundCommon bean (Phaseolus vulgaris L.) and soybean (Glycine max) both belong to the Phaseoleae tribe and share significant coding sequence homology. This suggests that the GeneChip® Soybean Genome Array (soybean GeneChip) may be used for gene expression studies using common bean.ResultsTo evaluate the utility of the soybean GeneChip for transcript profiling of common bean, we hybridized cRNAs purified from nodule, leaf, and root of common bean and soybean in triplicate to the soybean GeneChip. Initial data analysis showed a decreased sensitivity and accuracy of measuring differential gene expression in common bean cross-species hybridization (CSH) GeneChip data compared to that of soybean. We employed a method that masked putative probes targeting inter-species variable (ISV) regions between common bean and soybean. A masking signal intensity threshold was selected that optimized both sensitivity and accuracy of measuring differential gene expression. After masking for ISV regions, the number of differentially-expressed genes identified in common bean was increased by 2.8-fold reflecting increased sensitivity. Quantitative RT-PCR (qRT-PCR) analysis of 20 randomly selected genes and purine-ureide pathway genes demonstrated an increased accuracy of measuring differential gene expression after masking for ISV regions. We also evaluated masked probe frequency per probe set to gain insight into the sequence divergence pattern between common bean and soybean. The sequence divergence pattern analysis suggested that the genes for basic cellular functions and metabolism were highly conserved between soybean and common bean. Additionally, our results show that some classes of genes, particularly those associated with environmental adaptation, are highly divergent.ConclusionsThe soybean GeneChip is a suitable cross-species platform for transcript profiling in common bean when used in combination with the masking protocol described. In addition to transcript profiling, CSH of the GeneChip in combination with masking probes in the ISV regions can be used for comparative ecological and/or evolutionary genomics studies.

Variation and inheritance of iron reductase activity in the roots of common bean (Phaseolus vulgaris L.) and association with seed iron accumulation QTL

BackgroundIron deficiency anemia is a global problem which often affects women and children of developing countries. Strategy I plants, such as common bean (Phaseolus vulgaris L.) take up iron through a process that involves an iron reduction mechanism in their roots; this reduction is required to convert ferric iron to ferrous iron. Root absorbed iron is critical for the iron nutrition of the plant, and for the delivery of iron to the shoot and ultimately the seeds. The objectives of this study were to determine the variability and inheritance for iron reductase activity in a range of genotypes and in a low × high seed iron cross (DOR364 × G19833), to identify quantitative trait loci (QTL) for this trait, and to assess possible associations with seed iron levels.ResultsThe experiments were carried out with hydroponically grown plants provided different amounts of iron varying between 0 and 20 μM Fe(III)-EDDHA. The parents, DOR364 and G19833, plus 13 other cultivated or wild beans, were found to differ in iron reductase activity. Based on these initial experiments, two growth conditions (iron limited and iron sufficient) were selected as treatments for evaluating the DOR364 × G19833 recombinant inbred lines. A single major QTL was found for iron reductase activity under iron-limited conditions (1 μM Fe) on linkage group b02 and another major QTL was found under iron sufficient conditions (15 μM Fe) on linkage group b11. Associations between the b11 QTL were found with several QTL for seed iron.ConclusionsGenes conditioning iron reductase activity in iron sufficient bean plants appear to be associated with genes contributing to seed iron accumulation. Markers for bean iron reductase (FRO) homologues were found with in silico mapping based on common bean synteny with soybean and Medicago truncatula on b06 and b07; however, neither locus aligned with the QTL for iron reductase activity. In summary, the QTL for iron reductase activity under iron limited conditions may be useful in environments where beans are grown in alkaline soils, while the QTL for iron reductase under sufficiency conditions may be useful for selecting for enhanced seed nutritional quality.

Anatomical root variations in response to water deficit: wild and domesticated common bean (Phaseolus vulgaris L)

Root anatomical responses to water deficit are diverse and regulation of water uptake strongly depends on plant anatomy. The ancestors of common bean (Phaseolus vulgaris L.) cultivars are the wild common beans. Because wild beans adapt and survive well in the natural environment, it is hypothesized that wild common bean roots are less affected than those of domesticated beans at low substrate water potential (ψW). A wild common bean accession from Chihuahua Mexico and cv. Bayomex were studied. Seedlings with a mean root length between 3 and 4 cm were maintained for 24 h in vermiculite at ψW of -0.03 (well hydrated), -0.65, -1.48 and -2.35 MPa (partially dry). Ten anatomical characteristics of differentiation and cell division in root regions were evaluated. Thickness of epidermis and protoderm diminished similarly in wild and domesticated beans growing at low substrate ψW (between -0.65 and -2.35 MPa). At the same time, parenchymatic cell area diminished by 71 % in the domesticated variety, but by only 32 % in the wild bean at -2.35 MPa. The number of cells in the cortex and the thickness of the xylem wall increased in both wild and domesticated beans at low substrate ψW; nevertheless, the effect was significantly lower in the wild bean. The number of xylem vessels increased in the cultivar (up to 40 %) while in the wild bean it decreased (up to 33 %). The diameter of xylem vessels and transverse root area diminished (15 and 57 %, respectively) in the cultivar, but in the wild common bean were not affected. Anatomical root characteristics and their modifications in both differentiation and cell division in root regions demonstrated that the wild bean reacted quite differently to substrate ψW than the domesticated common bean.

Biochemical and molecular characterization of PvPAP3, a novel purple acid phosphatase isolated from common bean enhancing extracellular ATP utilization.

Purple acid phosphatases (PAPs) play diverse physiological roles in plants. In this study, we purified a novel PAP, PvPAP3, from the roots of common bean (Phaseolus vulgaris) grown under phosphate (Pi) starvation. PvPAP3 was identified as a 34-kD monomer acting on the specific substrate, ATP, with a broad pH range and a high heat stability. The activity of PvPAP3 was insensitive to tartrate, indicating that PvPAP3 is a PAP-like protein. Amino acid sequence alignment and phylogenetic analysis suggest that PvPAP3 belongs to the group of plant PAPs with low molecular mass. Transient expression of 35S:PvPAP3-green fluorescent protein in onion (Allium cepa) epidermal cells verified that it might anchor on plasma membrane and be secreted into apoplast. Pi starvation led to induction of PvPAP3 expression in both leaves and roots of common bean, and expression of PvPAP3 was strictly dependent on phosphorus (P) availability and duration of Pi starvation. Furthermore, induction of PvPAP3 expression was more rapid and higher in a P-efficient genotype, G19833, than in a P-inefficient genotype, DOR364, suggesting possible roles of PvPAP3 in P efficiency in bean. In vivo analysis using a transgenic hairy root system of common bean showed that both growth and P uptake of bean hairy roots from the PvPAP3 overexpression transgenic lines were significantly enhanced when ATP was supplied as the sole external P source. Taken together, our results suggest that PvPAP3 is a novel PAP that might function in the adaptation of common bean to P deficiency, possibly through enhancing utilization of extracellular ATP as a P source.

Effect of phosphorus limiting on phytase activity, proton efflux and oxygen consumption by nodulated- roots of common bean ( Phaseolus vulgaris )

This work intended to measure the nodulated-roots oxygen consumption, proton efflux and phytase activity in 2 lines of common bean (Phaseolus vulgaris) (115, 147) at 2 levels of P supply. Rooted seedlings were inoculated with Rhizobium tropici CIAT 899 in hydroaeroponic cultivation under glasshouse. Phosphorus was supplied as KH2PO4 at 15 and 250 imol pl-1 week-1 (15P and 250P, respectively). Our results showed that plant growth nodulation and symbiotic nitrogen fixation were significantly affected by P limiting (15P) for the both lines, but this adverse effect was more pronounced in 147 than in 115. For the both lines, the phytase activity, higher in roots than in nodules, was significantly increased by P limiting, but 115 maintained higher values as compared to 147 line. In cotyledons, the phytase activity was higher in 115 than in 147. Phosphorus shortage increased the cumulated proton release only in 115, whereas it was lowered for 147. In this line, the proton release was linked to symbiotic nitrogen fixation. Under 15P, the proton efflux per unit of nodulated-root biomass was 25% greater for 115 than 147, suggesting that under P limitation, proton efflux may constitute an efficient way to increase P uptake in the tolerant line (115). 15P increased significantly nodulated-root O2 consumption per g nodule DW and nodule conductance, but to a higher extent in 147. As a whole, bean plants at P-deficient conditions increased the activity of phytases and proton efflux, thus maintaining the oxygen diffusion in nodules. This may represent an adaptive mechanism for N2- fixing legumes to respond to P deficiency, by increasing the utilisation and the uptake of phosphorus for symbiotic nitrogen fixation.

Assess suitability of hydroaeroponic culture to establish tripartite symbiosis between different AMF species, beans, and rhizobia.

BackgroundLike other species of the Phaseoleae tribe, common bean (Phaseolus vulgaris L.) has the potential to establish symbiosis with rhizobia and to fix the atmospheric dinitrogen (N2) for its N nutrition. Common bean has also the potential to establish symbiosis with arbuscular mycorrhizal fungi (AMF) that improves the uptake of low mobile nutrients such as phosphorus, from the soil. Both rhizobial and mycorrhizal symbioses can act synergistically in benefits on plant.ResultsThe tripartite symbiosis of common bean with rhizobia and arbuscular mycorrhizal fungi (AMF) was assessed in hydroaeroponic culture with common bean (Phaseolus vulgaris L.), by comparing the effects of three fungi spp. on growth, nodulation and mycorrhization of the roots under sufficient versus deficient P supplies, after transfer from initial sand culture. Although Glomus intraradices Schenck & Smith colonized intensely the roots of common bean in both sand and hydroaeroponic cultures, Gigaspora rosea Nicolson & Schenck only established well under sand culture conditions, and no root-colonization was found with Acaulospora mellea Spain & Schenck under either culture conditions. Interestingly, mycorrhization by Glomus was also obtained by contact with mycorrhized Stylosanthes guianensis (Aubl.) sw in sand culture under deficient P before transfer into hydroaeroponic culture. The effect of bean genotype on both rhizobial and mycorrhizal symbioses with Glomus was subsequently assessed with the common bean recombinant inbreed line 7, 28, 83, 115 and 147, and the cultivar Flamingo. Significant differences among colonization and nodulation of the roots and growth among genotypes were found.ConclusionThe hydroaeroponic culture is a valuable tool for further scrutinizing the physiological interactions and nutrient partitioning within the tripartite symbiosis.

Application of Pluronic gel to the study of root-knot nematode behaviour

Abstract Pluronic F-127 is a stable, non-toxic, copolymer that forms a gel at room temperature and a liquid at 15°C when the concentration is 20-30%. Root-knot nematode (Meloidogyne spp.) second-stage juveniles can move freely through the gel and display attraction toward roots of tomato, Medicago truncatula, common bean and Arabidopsis. The excellent clarity of the gel allows examination of behavioural changes in the nematode as it reaches and infects its host. Attraction assays showed that Meloidogyne javanica and M. incognita move to roots much more rapidly than does M. hapla. Nematodes form aggregates when in contact with root tips suggesting that the presence of a signal from the nematode or from the root is involved in the attraction. Nematodes suspended in Pluronic gel without roots aggregate into balls after 1 to 2 days. A coverslip placed on the gel accelerates and serves as a focus for the aggregation, suggesting that lower oxygen, or perhaps a volatile attractant, is involved in this behaviour. These observations demonstrate that Pluronic gel is a useful medium for dissecting attraction of root-knot nematodes to their hosts and for studying additional aspects of their behaviour.

Essential role of MYB transcription factor: PvPHR1 and microRNA: PvmiR399 in phosphorus‐deficiency signalling in common bean roots

Phosphorus (P), an essential element for plants, is one of the most limiting nutrients for plant growth. A few transcription factor (TF) genes involved in P-starvation signalling have been characterized for Arabidopsis thaliana and rice. Crop production of common bean (Phaseolus vulgaris L.), the most important legume for human consumption, is often limited by low P in the soil. Despite its agronomic importance, nothing is known about transcriptional regulation in P-deficient bean plants. We functionally characterized the P-deficiency-induced MYB TF TC3604 (Dana Farber Cancer Institute, Common Bean Gene Index v.2.0), ortholog to AtPHR1 (PvPHR1). For its study, we applied RNAi technology in bean composite plants. PvPHR1 is a positive regulator of genes implicated in P transport, remobilization and homeostasis. Although there are no reports on the regulatory roles of microRNAs (miRNA) in bean, we demonstrated that PvmiR399 is an essential component of the PvPHR1 signalling pathway. The analysis of DICER-like1 (PvDCL1) silenced bean composite plants suppressed for accumulation of PvmiR399 and other miRNAs suggested that miR399 is a negative regulator of the ubiquitin E2 conjugase: PvPHO2 expression. Our results set the basis for understanding the signalling for P-starvation responses in common bean and may contribute to crop improvement.

Breeding for Drought Resistance in Common Bean: In Vitro Assay for Root Osmotic Potential

ABSTRACT Initial and adjusted osmotic potentials of separate leaf- and root-derived calli were assessed in vitro for two teparies (Phaseolus acutifolius A. Czray) and a common bean (P. vulgaris L.) cultivar differing in physiological and morpho-physiological reactions of intact plants to water stress. Unstressed leaf- and root-derived calli of common bean (CB) had higher preexisting osmotic potential (ψs) and relative growth rate (RGR) but lower dry-matter percent (DM) than those of tepary bean (TB) lines when grown on basal Murashige and Skoog medium (–0.30 MPa). Exposure to water stress (−0.58 MPa) imposed by adding polyethylene glycol (10% [w/v], PEG-10,000) did not affect these parameters for leaf-derived callus of CB and both leaf- and root-derived calli of TB. However, root-derived callus of CB showed decreased ψs that was maintained after transferring stressed callus onto medium lacking PEG for recovery from water stress. Nevertheless, its RGR decreased and DM increased. Stressed plants of both CB and TB grown in the greenhouse manifested significant reduction in leaf, stem, and root dry mass and leaf area. However, the magnitude of mass reduction was especially high for CB stem, leaf, and root. While TB root system penetrated deeper in the soil profile, CB root depth did not significantly change in response to drought. In addition, substantial CB root mass was detected in the top 10 to 20 cm of the soil profile. Stomata conductance and transpiration rate were higher for CB than TB under both well-watered and water-stress conditions. Significant relative water content reduction was detected only in CB. In spite of water stress, soil moisture was high around roots of CB compared with TB, suggesting lesser efficiency of CB root system to extract water. Stressed CB grown in the production field did not produce seed yield whereas TB gave 60 to 64% of its potential dry seed yield. In vitro assay for root osmotic potential should be useful in improving drought resistance in common bean.

Proton release by nodulated roots varies among common bean genotypes (Phaseolus vulgaris) under phosphorus deficiency

AbstractResponses of proton release to phosphorus (P) availability by nodulated roots of common bean (Phaseolus vulgaris L.) were investigated for lines BAT 477 and CocoT, inoculated with Rhizobium tropici CIAT 899 in hydroaeroponic culture under glasshouse conditions. Phosphorus was supplied as KH2PO4 at 15 and 60 μmol plant–1 week–1 (15P and 60P). Proton release was higher for BAT 477 than for CocoT under both P supplies. However, it was higher for 60P than 15P, whatever the line. The ratio of proton release per unit biomass of nodulated root was higher for BAT 477 than for CocoT, independent of P deficiency. Proton release was correlated with the nodulated‐root respiration for both genotypes and with the nodule respiration linked with nitrogen fixation for CocoT. Thus, the nodulation was more limited by 15P than root and shoot growth and more in CocoT than in BAT 477. It is concluded that independent of symbiotic N2 fixation, proton release was higher in BAT 477 than in CocoT and that the nodulated legume releases a substantial amount of protons into its rhizosphere that is correlated with its nitrogen fixation that eventually depends upon the nodule permeability to O2 diffusion.