Seminal and Nodal Roots of Barley Differ in Anatomy, Proteome and Nitrate Uptake Capacity.

The root system of barley plants is composed of embryogenic, seminal roots as well as lateral and nodal roots that are formed postembryonically from seminal roots and from the basal part of shoots, respectively. Due to their distinct developmental origin, seminal and nodal roots may differ in function during plant development; however, a clear comparison between these two root types has not yet been undertaken. In this study, anatomical, proteomic and physiological traits were compared between seminal and nodal roots of similar developmental stages. Nodal roots have larger diameter, larger metaxylem area and a larger number of metaxylem vessels than seminal roots. Proteome profiling uncovered a set of root-type-specific proteins, including proteins related to the cell wall and cytoskeleton organization, which could potentially be implicated with differential metaxylem development. We also found that nodal roots have higher levels of auxin, which is known to trigger metaxylem development. At millimolar nitrate supply, nodal roots had approximately 2-fold higher nitrate uptake and root-to-shoot translocation capacities than seminal roots, whereas no differences were found at micromolar nitrate supply. Since these marked differences were not reflected by the transcript levels of low-affinity nitrate transporter genes, we hypothesize that the larger metaxylem volume of nodal roots enhances predominantly the low-affinity uptake and translocation capacities of nutrients that are transported with the bulk flow of water, like nitrate.

Biochemical and Anatomical Investigation of Sesbania herbacea (Mill.) McVaugh Nodules Grown under Flooded and Non-Flooded Conditions.

Sesbania herbacea, a native North American fast-growing legume, thrives in wet and waterlogged conditions. This legume enters into symbiotic association with rhizobia, resulting in the formation of nitrogen-fixing nodules on the roots. A flooding-induced anaerobic environment imposes a challenge for the survival of rhizobia and negatively impacts nodulation. Very little information is available on how S. herbacea is able to thrive and efficiently fix N2 in flooded conditions. In this study, we found that Sesbania plants grown under flooded conditions were significantly taller, produced more biomass, and formed more nodules when compared to plants grown on dry land. Transmission electron microscopy of Sesbania nodules revealed bacteroids from flooded nodules contained prominent polyhydroxybutyrate crystals, which were absent in non-flooded nodules. Gas and ion chromatography mass spectrometry analysis of nodule metabolites revealed a marked decrease in asparagine and an increase in the levels of gamma aminobutyric acid in flooded nodules. 2-D gel electrophoresis of nodule bacteroid proteins revealed flooding-induced changes in their protein profiles. Several of the bacteroid proteins that were prominent in flooded nodules were identified by mass spectrometry to be members of the ABC transporter family. The activities of several key enzymes involved in nitrogen metabolism was altered in Sesbania flooded nodules. Aspartate aminotransferase (AspAT), an enzyme with a vital role in the assimilation of reduced nitrogen, was dramatically elevated in flooded nodules. The results of our study highlight the potential of S. herbacea as a green manure and sheds light on the morphological, structural, and biochemical adaptations that enable S. herbacea to thrive and efficiently fix N2 in flooded conditions.

Three-dimensional reconstruction of soybean nodules provides an update on vascular structure.

PREMISE OF THE STUDY: In many cases, the functioning of a biological system cannot be correctly understood if its physical anatomy is incorrectly described. Accurate knowledge of the anatomy of soybean [Glycine max (L.) Merril] nodules and its connection with the root vasculature is important for understanding its function in supplying the plant with nitrogenous compounds. Previous two-dimensional anatomical observations of soybean nodules led to the assumption that vascular bundles terminate within the cortex of the nodule and that a single vascular bundle connects the nodule to the root. We wanted to see whether these anatomical assumptions would be verified by digitally reconstructing soybean nodules in three dimensions. METHODS: Nodules were dehydrated, embedded in paraffin, and cut into 15 µm thick sections. Over 200 serial sections were stained with safranin and fast green, and then photographed using light microscopy. Images were digitally cleared, aligned, and assembled into a three-dimensional (3D) volume using the Adobe program After Effects. KEY RESULTS: In many cases, vascular bundles had a continuous connection around the nodules. The 3D reconstruction also revealed a dual vascular connection originating in the nodule and leading to the root in 22 of the 24 nodules. Of the 22 dual connections, 11 maintained two separate vascular bundles into the root with independent connections to the root vasculature. CONCLUSIONS: A more robust and complex anatomical pathway for vascular transport between nodules and root xylem in soybean plants is indicated by these observations and will contribute to a better understanding of the symbiotic relationship between soybean plants and nitrogen-fixing bacteria within the nodules.

Proteomic and ecophysiological responses of soybean (Glycine max L.) root nodules to Pb and hg stress.

BACKGROUND: Lead (Pb) and mercury (Hg) are persistent hazardous metals in industrially polluted soils which can be toxic in low quantities. Metal toxicity can cause changes at cellular and molecular level which should be studied for better understanding of tolerance mechanism in plants. Soybean (Glycine max L.) is an important oilseed crop of the world including India. Indian soils growing soybean are often contaminated by Pb and Hg. The aim of this study was to explore how soybean root nodule responds to Pb and Hg through proteomic and ecophysiological alterations in order to enhance tolerance to metal stress. RESULTS: Soybean plants were exposed to Pb (30 ppm PbCl2) and Hg (0.5 ppm HgCl2) to study histological, histochemical, biochemical and molecular response of N2-fixing symbiotic nodules. Both Pb and Hg treatment increased the level of oxidative stress in leaves and nodules. Chlorosis in leaves and morphological/anatomical changes in nodules were observed. Activities of ascorbate peroxidase, glutathione reductase and catalase were also modulated. Significant changes were observed in abundance of 76 proteins by Pb and Hg. Pb and Hg influenced abundance of 33 proteins (17 up and 16 down) and 43 proteins (33 up and 10 down), respectively. MS/MS ion search identified 55 proteins which were functionally associated with numerous cellular functions. Six crucial proteins namely catalase (CAT), allene oxide synthase (AOS), glutathione S-transferase (GST), calcineurin B like (CBL), calmodulin like (CML) and rapid alkalinisation factor (RAF) were selected for transcript abundance estimation. The qRT-PCR based real time expression exhibited a positive correlation with proteomics expression except for GST and RAF. CONCLUSION: Soybean root nodule responds to metal stress by increased abundance of defence, development and repair related proteins. An efficient proteomic modulation might lead to metal-induced stress tolerance in N2-fixing nodules. Although concentrations of Pb and Hg used in the study cannot be considered equimolar, yet Hg seems to induce more changes in nodule proteomic profile, and higher damage to both bacteroides and root anatomy.

Selection of Bradyrhizobium or Ensifer symbionts by the native Indian caesalpinioid legume Chamaecrista pumila depends on soil pH and other edaphic and climatic factors.

Nodules of Chamaecrista pumila growing in several locations in India were sampled for anatomical studies and for characterization of their rhizobial microsymbionts. Regardless of their region of origin, the nodules were indeterminate with their bacteroids contained within symbiosomes which were surrounded by pectin. More than 150 strains were isolated from alkaline soils from the Thar Desert (Rajasthan), wet-acidic soils of Shillong (Meghalaya), and from trap experiments using soils from four other states with different agro-ecological regions. Molecular phylogenetic analysis based on five housekeeping (rrs, recA, glnII, dnaK andatpD) and two symbiotic (nodA and nifH) genes was performed for selected strains. Chamaecrista pumila was shown to be nodulated by niche-specific diverse strains of either Ensifer or Bradyrhizobium in alkaline (Thar Desert) to neutral (Tamil Nadu) soils and only Bradyrhizobium strains in acidic (Shillong) soils. Concatenated core gene phylogenies showed four novel Ensifer-MLSA types and nine Bradyrhizobium-MLSA types. Genetically diverse Ensifer strains harbored similar sym genes which were novel. In contrast, significant symbiotic diversity was observed in the Bradyrhizobium strains. The C. pumila strains cross-nodulated Vigna radiata and some wild papilionoid and mimosoid legumes. It is suggested that soil pH and moisture level played important roles in structuring the C. pumila microsymbiont community.

Functional characterisation of brassinosteroid receptor MtBRI1 in Medicago truncatula.

Brassinosteroids are phytohormones involved in plant development and physiological processes. Brassinosteroids Insensitive 1 (BRI1) is required for BR perception and initiation of subsequent signal transduction in Arabidopsis. In this study, the orthologue of BRI1 in the model legume species Medicago truncatula, MtBRI1, was identified and characterised. Three allelic Tnt1 insertion mutants, mtbri1-1, mtbri1-2, and mtbri1-3, were obtained from the M. truncatula Tnt1 insertion population. mtbri1 mutants displayed characteristic bri1 mutant phenotypes: extreme dwarfness, dark green curled leaves, short primary roots, less lateral roots, and insensitive to exogenous brassinolide (BL). Moreover, mtbri1 mutants show decreased total nodule number and defects in nitrogen fixation. MtBRI1 is able to complement an Arabidopsis BRI1 mutant, bri1-5. Similar to the interaction of BRI1 and BAK1 in Arabidopsis, MtBRI1 interacts with MtSERK1 in vivo. Global gene expression profiling revealed that the expression of BR biosynthesis genes and SAUR genes are significantly altered in mtbri1 mutants. MapMan analysis indicated that genes involved in signaling, hormone, cell wall, and biotic stress responses are over-represented in differentially expressed genes. Taken together, the results indicate that MtBRI1 is the BR receptor in M. truncatula and that BR signaling may play a conserved role in balancing plant growth and defenses.

The presence of nodules on legume root systems can alter phenotypic plasticity in response to internal nitrogen independent of nitrogen fixation.

All higher plants show developmental plasticity in response to the availability of nitrogen (N) in the soil. In legumes, N starvation causes the formation of root nodules, where symbiotic rhizobacteria fix atmospheric N2 for the host in exchange for fixed carbon (C) from the shoot. Here, we tested whether plastic responses to internal [N] of legumes are altered by their symbionts. Glasshouse experiments compared root phenotypes of three legumes, Medicago truncatula, Medicago sativa and Trifolium subterraneum, inoculated with their compatible symbiont partners and grown under four nitrate levels. In addition, six strains of rhizobia, differing in their ability to fix N2 in M. truncatula, were compared to test if plastic responses to internal [N] were dependent on the rhizobia or N2 -fixing capability of the nodules. We found that the presence of rhizobia affected phenotypic plasticity of the legumes to internal [N], particularly in root length and root mass ratio (RMR), in a plant species-dependent way. While root length responses of M. truncatula to internal [N] were dependent on the ability of rhizobial symbionts to fix N2 , RMR response to internal [N] was dependent only on initiation of nodules, irrespective of N2 -fixing ability of the rhizobia strains.

Deletions of the SACPD-C locus elevate seed stearic acid levels but also result in fatty acid and morphological alterations in nitrogen fixing nodules.

BACKGROUND: Soybean (Glycine max) seeds are the primary source of edible oil in the United States. Despite its widespread utility, soybean oil is oxidatively unstable. Until recently, the majority of soybean oil underwent chemical hydrogenation, a process which also generates trans fats. An alternative to chemical hydrogenation is genetic modification of seed oil through identification and introgression of mutant alleles. One target for improvement is the elevation of a saturated fat with no negative cardiovascular impacts, stearic acid, which typically constitutes a minute portion of seed oil (~3%). RESULTS: We examined radiation induced soybean mutants with moderately increased stearic acid (10-15% of seed oil, ~3-5 X the levels in wild-type soybean seeds) via comparative whole genome hybridization and genetic analysis. The deletion of one SACPD isoform encoding gene (SACPD-C) was perfectly correlated with moderate elevation of seed stearic acid content. However, SACPD-C deletion lines were also found to have altered nodule fatty acid composition and grossly altered morphology. Despite these defects, overall nodule accumulation and nitrogen fixation were unaffected, at least under laboratory conditions. CONCLUSIONS: Although no yield penalty has been reported for moderate elevated seed stearic acid content in soybean seeds, our results demonstrate that genetic alteration of seed traits can have unforeseen pleiotropic consequences. We have identified a role for fatty acid biosynthesis, and SACPD activity in particular, in the establishment and maintenance of symbiotic nitrogen fixation.

Morphological and genetic changes induced by excess Zn in roots of Medicago truncatula A17 and a Zn accumulating mutant.

BACKGROUND: Nutrient fluxes associated with legume-rhizobia symbioses are poorly understood and little is known regarding the influence of abiotic stresses on development and maintenance of N-fixing nodules and root system architecture (RSA). We examined effects of Zn on nodule development and structure, root architecture, and expression of nodulation-related miRNAs in Medicago truncatula and the mutant, raz (requires additional Zn). FINDINGS: Excess Zn increased root and shoot associated Zn in both genotypes, however, raz plants had lower root associated Zn than WT plants. Roots of raz plants exposed to excess Zn had less volume, surface area, and total length compared to WT plants. Raz plants had lower lateral root number than WT plants. Excess Zn was found to increase root diameter in both genotypes. The Mn Translocation Factor (TfMn) increased in response to Zn in both genotypes; this was more pronounced in raz plants. TfZn was higher in raz plants and reduced in both genotypes in response to Zn. Nodulation was not influenced by Zn treatment or plant genotype. MicroRNA166 was upregulated under excess Zn in WT plants. CONCLUSIONS: Neither the raz mutation nor Zn treatment affected nodulation, however, raz plants had altered RSA compared with WT and responded differently to Zn, implying the mutation potentially modulates RSA responses to Zn but doesn't play a direct role in nodulation. MicroRNA166 was significantly induced in WT plants by excess Zn, warranting further investigation into the potential role it plays in controlling RSA.

A functional-structural modelling approach to autoregulation of nodulation.

BACKGROUND AND AIMS: Autoregulation of nodulation is a long-distance shoot-root signalling regulatory system that regulates nodule meristem proliferation in legume plants. However, due to the intricacy and subtleness of the signalling nature in plants, molecular and biochemical details underlying mechanisms of autoregulation of nodulation remain largely unknown. The purpose of this study is to use functional-structural plant modelling to investigate the complexity of this signalling system. There are two major challenges to be met: modelling the 3D architecture of legume roots with nodulation and co-ordinating signalling-developmental processes with various rates. METHODS: Soybean (Glycine max) was chosen as the target legume. Its root system was observed to capture lateral root branching and nodule distribution patterns. L-studio, a software tool supporting context-sensitive L-system modelling, was used for the construction of the architectural model and integration with the internal signalling. KEY RESULTS: A branching pattern with regular radial angles was found between soybean lateral roots, from which a root mapping method was developed to characterize the laterals. Nodules were mapped based on 'nodulation section' to reveal nodule distribution. A root elongation algorithm was then developed for simulation of root development. Based on the use of standard sub-modules, a synchronization algorithm was developed to co-ordinate multi-rate signalling and developmental processes. CONCLUSIONS: The modelling methods developed here not only allow recreation of legume root architecture with lateral branching and nodulation details, but also enable parameterization of internal signalling to produce different regulation results. This provides the basis for using virtual experiments to help in investigating the signalling mechanisms at work.

Genetic dissection of nitrogen nutrition in pea through a QTL approach of root, nodule, and shoot variability.

Pea (Pisum sativum L.) is the third most important grain legume worldwide, and the increasing demand for protein-rich raw material has led to a great interest in this crop as a protein source. Seed yield and protein content in crops are strongly determined by nitrogen (N) nutrition, which in legumes relies on two complementary pathways: absorption by roots of soil mineral nitrogen, and fixation in nodules of atmospheric dinitrogen through the plant-Rhizobium symbiosis. This study assessed the potential of naturally occurring genetic variability of nodulated root structure and functioning traits to improve N nutrition in pea. Glasshouse and field experiments were performed on seven pea genotypes and on the 'Cameor' x 'Ballet' population of recombinant inbred lines selected on the basis of parental contrast for root and nodule traits. Significant variation was observed for most traits, which were obtained from non-destructive kinetic measurements of nodulated root and shoot in pouches, root and shoot image analysis, (15)N quantification, or seed yield and protein content determination. A significant positive relationship was found between nodule establishment and root system growth, both among the seven genotypes and the RIL population. Moreover, several quantitative trait loci for root or nodule traits and seed N accumulation were mapped in similar locations, highlighting the possibility of breeding new pea cultivars with increased root system size, sustained nodule number, and improved N nutrition. The impact on both root or nodule traits and N nutrition of the genomic regions of the major developmental genes Le and Af was also underlined.

[Root morphology and nodule traits of two soybean varieties on alfisol and mollisol].

A pot experiment was conducted with two soybean varieties (Di2003-1 and Hefeng25) and two main soil types (alfisol and mollisol) in Heilongjiang Province to study the interactive effects of variety and soil type on the root morphology and nodule traits of soybean plants. Root samples were collected at the fifth trifoliolate stage, bloom-beginning stage, pod-beginning stage, full pod stage, seed-beginning stage, full seed stage, and full maturity stage for the analysis of root dry mass, root length, root surface area, average root diameter, nodule number, nodule fresh mass, and mass per nodule. The results showed that soil type had significant effects on the root morphology and nodule traits of test varieties. In alfisol, the root dry mass, root length, and root surface area of test varieties were greater at the fifth trifoliolate stage and bloom-beginning stages while smaller after seed-beginning stage, compared with those in mollisol, and the average root diameter after seed-beginning stage was higher in alfisol than that in mollisol. Soil type did not affect root-shoot ratio. The nodule number after seed-beginning stage was much smaller in alfisol than in mollisol, while the nodule fresh mass and the mass per nodule were in adverse. The root traits in the two soil types varied with soybean variety. Compared with that of Hefeng25, the response of root traits of Di2003-1 to soil type was more sensitive. The interactive effects of variety and soil type on the root morphology and nodule traits of soybean plants were more significant at full seed stage.

The model legume Medicago truncatula A17 is poorly matched for N2 fixation with the sequenced microsymbiont Sinorhizobium meliloti 1021.

Medicago truncatula (barrel medic) A17 is currently being sequenced as a model legume, complementing the sequenced root nodule bacterial strain Sinorhizobium meliloti 1021 (Sm1021). In this study, the effectiveness of the Sm1021-M. truncatula symbiosis at fixing N(2) was evaluated. N(2) fixation effectiveness was examined with eight Medicago species and three accessions of M. truncatula with Sm1021 and two other Sinorhizobium strains. Plant shoot dry weights, plant nitrogen content and nodule distribution, morphology and number were analysed. Compared with nitrogen-fed controls, Sm1021 was ineffective or partially effective on all hosts tested (excluding M. sativa), as measured by reduced dry weights and shoot N content. Against an effective strain, Sm1021 on M. truncatula accessions produced more nodules, which were small, pale, more widely distributed on the root system and with fewer infected cells. The Sm1021-M. truncatula symbiosis is poorly matched for N(2) fixation and the strain could possess broader N(2) fixation deficiencies. A possible origin for this reduction in effectiveness is discussed. An alternative sequenced strain, effective at N(2) fixation on M. truncatula A17, is Sinorhizobium medicae WSM419.

Developmental changes in peanut root structure during root growth and root-structure modification by nodulation.

BACKGROUND AND AIMS: Basic information about the root and root nodule structure of leguminous crop plants is incomplete, with many aspects remaining unresolved. Peanut (Arachis hypogaea) forms root nodules in a unique process. Structures of various peanut root types were studied with emphasis on insufficiently characterized lateral roots, changes in roots during their ontogenesis and root modification by nodule formation. METHODS: Peanut plants were grown in the field, in vermiculite or in filter paper. The taproot, first-order and second-order lateral roots and root nodules were analysed using bright-field and fluorescence microscopy with hand sections and resin sections. KEY RESULTS: Three root categories were recognized. The primary seminal root was thick, exhibiting early and intensive secondary thickening mainly on its base. It was tetrarch and contained broad pith. First-order lateral roots were long and thin, with limited secondary thickening; they contained no pith. Particularly different were second- and higher-order lateral roots, which were anatomically simple and thin, with little or no secondary growth. Unusual wall ingrowths were visible in the cells of the central part of the cortex in the first-order and second-order lateral roots. The nodule body was formed at the junction of the primary and lateral roots by the activity of proliferating cells derived originally from the pericycle. CONCLUSIONS: Two morphologically and anatomically distinct types of lateral roots were recognized: long, first-order lateral roots, forming the skeleton of the root system, and thin and short second- and higher-order lateral roots, with an incomplete second state of endodermal development, which might be classified as peanut 'feeder roots'. Formation of root nodules at the base of the lateral roots was the result of proliferating cell divisions derived originally from the pericycle.

Abnormal root and nodule vasculature in R50 (sym16), a pea nodulation mutant which accumulates cytokinins.

BACKGROUND AND AIMS: R50 (sym16) is a pea nodulation mutant with fewer and shorter lateral roots (LR), fewer nodules and high levels of cytokinins (CK). Because a link exists between CK imbalance and abnormal vasculature, the vasculature of the primary root (PR) and LR of R50 was studied and it was compared with that of the wild-type 'Sparkle'. Also nodule vasculature was investigated to correlate R50 low nodulation phenotype with CK accumulation. METHODS: PR and first-order LR were hand-sectioned transversely in different locations and at different ages. Vascular poles were counted and root and stele diameters measured. To evaluate LR primordia number, roots were cleared. Nodules obtained from inoculated plants were either fixed and sectioned or cleared; numbers of vascular strands and of tracheary elements in the strands were counted. KEY RESULTS: 'Sparkle' PR is triarch, whereas that of R50 can be triarch, tetrarch or pentarch. Furthermore, as the R50 roots developed, supernumerary vascular strands appeared but, as they aged, the new growth of more roots displayed the triarch pattern. LR vasculature differed from that of PR: whereas 'Sparkle' LR had three or four poles, those of R50 had two or three. No differences in PR or PR stele diameters existed between the two lines. Whereas 'Sparkle' nodules had two vascular strands, most R50 nodules possessed three; however, because R50 nodules were variable in size, their vasculature was highly diverse in terms of strand length. A strong correlation was found between nodule length and number of tracheary elements in strands. CONCLUSIONS: R50 displays an additional number of vascular poles in its PR, a smaller number of vascular poles in its first-order LR and an altered vasculature in its nodules. It appears that these three characteristics are linked to the high levels of CKs that the mutant accumulates over its development.

Nitrogen fixation by white lupin under phosphorus deficiency.

BACKGROUND AND AIMS: White lupin is highly adapted to growth in a low-P environment. The objective of the present study was to evaluate whether white lupin grown under P-stress has adaptations in nodulation and N2 fixation that facilitate continued functioning. METHODS: Nodulated plants were grown in silica sand supplied with N-free nutrient solution containing 0 to 0.5 mm P. At 21 and 37 d after inoculation (DAI) growth, nodulation, P and N concentration, N2 fixation (15N2 uptake and H2 evolution), root/nodule net CO2 evolution and CO2 fixation (14CO2 uptake) were measured. Furthermore, at 21 DAI in-vitro activities and transcript abundance of key enzymes of the C and N metabolism in nodules were determined. Moreover, nodulation in cluster root zones was evaluated. KEY RESULTS: Treatment without P led to a lower P concentration in shoots, roots, and nodules. In both treatments, with or without P, the P concentration in nodules was greater than that in the other organs. At 21 DAI nitrogen fixation rates did not differ between treatments and the plants displayed no symptoms of P or N deficiency on their shoots. Although nodule number at 21 DAI increased in response to P-deficiency, total nodule mass remained constant. Increased nodule number in P-deficient plants was associated with cluster root formation. A higher root/nodule CO2 fixation in the treatment without P led to a lower net CO2 release per unit fixed N, although the total CO2 released per unit fixed N was higher in the treatment without P. The higher CO2 fixation was correlated with increased transcript abundance and enzyme activities of phosphoenolpyruvate carboxylase and malate dehydrogenase in nodules. Between 21 and 37 DAI, shoots of plants grown without P developed symptoms of N- and P-deficiency. By 37 DAI the P concentration had decreased in all organs of the plants treated with no P. At 37 DAI, nitrogen fixation in the treatment without P had almost ceased. CONCLUSIONS: Enhanced nodulation in cluster root zones and increased potential for organic acid production in root nodules appear to contribute to white lupin's resilience to P-deficiency.

Silencing the flavonoid pathway in Medicago truncatula inhibits root nodule formation and prevents auxin transport regulation by rhizobia.

Legumes form symbioses with rhizobia, which initiate the development of a new plant organ, the nodule. Flavonoids have long been hypothesized to regulate nodule development through their action as auxin transport inhibitors, but genetic proof has been missing. To test this hypothesis, we used RNA interference to silence chalcone synthase (CHS), the enzyme that catalyzes the first committed step of the flavonoid pathway, in Medicago truncatula. Agrobacterium rhizogenes transformation was used to create hairy roots that showed strongly reduced CHS transcript levels and reduced levels of flavonoids in silenced roots. Flavonoid-deficient roots were unable to initiate nodules, even though normal root hair curling was observed. Nodule formation and flavonoid accumulation could be rescued by supplementation of plants with the precursor flavonoids naringenin and liquiritigenin. The flavonoid-deficient roots showed increased auxin transport compared with control roots. Inoculation with rhizobia reduced auxin transport in control roots after 24 h, similar to the action of the auxin transport inhibitor N-(1-naphthyl)phthalamic acid (NPA). Rhizobia were unable to reduce auxin transport in flavonoid-deficient roots, even though NPA inhibited auxin transport. Our results present genetic evidence that root flavonoids are necessary for nodule initiation in M. truncatula and suggest that they act as auxin transport regulators.

Morphological variation, histology, histochemistry and nutrient contents of Frankia root nodules in Casuarina equisetifolia.

Root nodules were collected from the existing plantation of Casuarina equisetifolia from three different agroclimatic zones of Tamil Nadu, India. Morphological variation, histology, histochemistry and nutrient contents of the nodules were studied. The nodules were found to be simple, dichotomously branched and coralloid. The stele in simple and branched nodules reached up to the tip of the nodules. Structure of Frankia nodules was described using histochemical methods to elucidate the nature of micro-symbiont within the nodules. The nodule consisted of a distinct periderm enclosing the cortex. The cortex showed patches of infected cells interspersed among the uninfected ones. Inside the cortex was a stele bound by an endodermis, which contained high amount of tannin. The micro-symbiont was observed in the nodule tisues in hyphal form and spore packed sporocorp was also seen in the cortex. The histochemical tests revealed the presence of proteins, insoluble polysaccharides and polyphenols.