Sites Of Lateral Root Emergence Research Articles

Crop plants transport irregularly shaped mineral particles from root to shoot: Tracking and quantifying

Mineral particles, ubiquitous in soils, influence crop plant growth by carrying nutrients and pollutants. While the uptake of dissolved mineral nutrients is well-established, the direct incorporation of irregular mineral particles into plants remains unclear. This study investigated the uptake and transport of kaolin particles, representative of minerals, by wheat and lettuce seedlings using hydroponic and soil cultures. Covalent labeling and advanced microscopy revealed that kaolin enters root steles at lateral root emergence sites, followed by transport to shoots. Fluorescent dyes and lanthanum (La)-labeled kaolin particles demonstrated that wheat surpassed lettuce in kaolin uptake in hydroponics, but both plants showed similar levels of particles in the shoots. Translocation factors (TFs) for kaolin were significantly higher in soil (0.089 for wheat, 0.039 for lettuce) compared to hydroponics (0.001 for wheat, 0.003 for lettuce). These findings provide compelling evidence for the direct uptake and transport of kaolin particles in crop plants. This opens new avenues for research on the interactions between plant and mineral particles, including other colloidal particles, in terrestrial ecosystems.

Bacillus thuringiensis translocation in citrus scion/rootstock combinations and binding of cry toxins with the Asian citrus psyllid gut receptors

Bacillus thuringiensis (Bt) can be effective in controlling insect pests either in spray products or by their toxins expressed in transgenic crops. The discovery of Bt as endophytic in citrus opened new perspectives for the control of Diaphorina citri Kuwayama, the vector of the bacteria Candidatus Liberibacter spp. associated with Huanglongbing. In this study, the endophytic translocation of Bt in citrus seedlings (small and large plants) and in different scion-rootstock combinations were investigated. We also assessed the lethal concentration of D. citri nymphs caused by systemic Bt in citrus and, moreover, the binding of Cry toxins with the Asian citrus psyllid (ACP) gut receptors. Endophytic translocation of Bt in citrus seedlings and nursery trees was confirmed by colony isolation, PCR detection, and Btk-GFP microscopy analyses, which demonstrated that, after Bt inoculation into the substrate, the bacteria enter through lateral root emergence sites and are localized in stem xylem vessel elements. Mean lethal concentrations for D. citri third instar nymphs were 4.92 × 104 and 2.19 × 104 spores mL−1 for S1450 and S1302 strains, respectively. Nymphal mortality induced by the bacteria was higher in citrus seedlings than in citrus nursery trees. All Cry-toxins tested were able to bind to D. citri brush border membrane vesicles, suggesting the potential pathogenicity of Bt strains against psyllid. This study clearly demonstrates the penetration and Bt translocation in the xylem of small and large citrus plants and the potential of the bacteria to cause ACP nymphs mortality that fed on shoots from Bt-treated citrus.

Ceratocystis fimbriata alters root system architecture and causes symptom development only in detached storage roots in Bayou Belle and Beauregard sweet potato

AbstractLittle is known about how the sweet potato (Ipomoea batatas) root system responds to Ceratocystis fimbriata in the growth substrate. Understanding whether the fungus affects the root system before harvest of storage roots (SR) could guide timing of management efforts. Cuttings of cultivars Bayou Belle and Beauregard, both previously considered susceptible to black rot, were in unamended or infested sand. At the onset of SR formation, effects on first‐ and second‐order lateral root (LR) length and number among and within treatments, cultivars and replicate experiments were recorded. First‐order LR length in inoculated Bayou Belle plants was 31% greater than in inoculated plants of cultivar Beauregard in Expt 1, but there were no differences in Expt 2. Second‐order LR length varied among, and within, cultivars, inoculum treatments and experiments in Expts 1 and 2. At 49 days after planting in Expt 1, only inoculated plants had necrotic lesions on stems in both cultivars, with more than twice as many recorded on Bayou Belle than Beauregard. SR had no lesions at harvest. Detached SR originating from inoculated treatments and stored in sampling bags for 25 days developed black rot lesions at either end of the SR (45% and 31% of total in Expt 1 and Expt 2, respectively) or elsewhere, including those centred on LR emergence sites or lenticels (55% and 69%, respectively). Exposing developing plants to inoculation may reveal differences in SR susceptibility not found when harvested SR are wounded and inoculated.

Colonization of Brachypodium distachyon by Gluconacetobacter diazotrophicus and its effect on plant growth promotion

Gluconacetobacter diazotrophicus is a nitrogen (N) fixing bacterium originally isolated from sugarcane in Brazil. Understanding the interactions between this bacterium and plants is important to exploit the nitrogen-fixing feature of G. diazotrophicus in different crops. Research was conducted to establish, monitor and optimize methods for introducing G. diazotrophicus into Brachypodium distachyon, a model for molecular research involving monocot plants. Colonization was studied in three plant growth media including liquid, semi-solid, and solid media. The effect of mineral nitrogen on colonization was also studied in each of these three media. The colonization of B. distachyon by G. diazotrophicus was analyzed by a histochemical β-glucuronidase (GUS) assay, PCR using G. diazotrophicus specific primers and bacterial re-isolation. Colonization was established in all tested plant growth conditions. The highest level of colonization was achieved in the liquid media without mineral nitrogen. Microscopic observations showed that G. diazotrophicus colonizes B. distachyon through the lateral root emergence sites, root hairs, and root tips. The colonization was found to be limited to root tissues and not found in other organs. These results indicate that B. distachyon roots are the primary organ of interaction and colonization by the bacterium. Plant fresh weight and lateral root numbers were significantly increased in inoculated plants indicating that G. diazotrophicus can stimulate growth of B. distachyon. This study indicates that B. distachyon is a useful model to study the mechanisms of G. diazotrophicus colonization and growth enhancement in monocot plants.

From the roots to the stem: unveiling pear root colonization and infection pathways by Erwinia amylovora.

Fire blight caused by Erwinia amylovora affects pome fruit worldwide, generating serious economic losses. Despite the abundant literature on E. amylovora infection mechanisms of aerial plant organs, root infection routes remain virtually unexplored. Assessing these infection pathways is necessary for a full understanding of the pathogen's ecology. Using the pathosystem Pyrus communis-E. amylovora and different experimental approaches including a green fluorescent protein transformant (GFP1) and epifluorescence microscopy (EFM) and laser confocal scanning microscopy (LCSM), we demonstrated the pathogen's ability to infect, colonize and invade pear roots and cause characteristic fire blight symptoms both in the aerial part and in the root system. Plant infections after soil irrigation with E. amylovora-contaminated water were favored by root damage, which agreed with EFM and LCSM observations. E. amylovora GFP1 cells formed aggregates/biofilms on root surfaces and invaded the cortex through wounds and sites of lateral root emergence. Sugars, sugar-alcohols and amino acids typically secreted by roots, favored the in vitro biofilm development by E. amylovora. Migration of E. amylovora cells to aerial tissues mainly occurred after xylem penetration. Overall, our findings revealed, for the first time, common root infection patterns between E. amylovora and well-known soil borne plant pathogens and endophytes.

Effective uptake of submicrometre plastics by crop plants via a crack-entry mode

Most microplastics are emitted, either directly or via the degradation of plastics, to the terrestrial environment and accumulate in large amounts in soils, representing a potential threat to terrestrial ecosystems. It is very important to evaluate the uptake of microplastics by crop plants because of the ubiquity of microplastics in wastewaters often used for agricultural irrigation worldwide. Here, we analyse the uptake of different microplastics by crop plants (wheat (Triticum aestivum) and lettuce (Lactuca sativa)) from treated wastewater in hydroponic cultures and in sand matrices or a sandy soil. Our results provide evidence in support of submicrometre- and micrometre-sized polystyrene and polymethylmethacrylate particles penetrating the stele of both species using the crack-entry mode at sites of lateral root emergence. This crack-entry pathway and features of the polymeric particles lead to the efficient uptake of submicrometre plastic. The plastic particles were subsequently transported from the roots to the shoots. Higher transpiration rates enhanced the uptake of plastic particles, showing that the transpirational pull was the main driving force of their movement. Our findings shed light on the modes of plastic particle interaction with plants and have implications for crops grown in fields contaminated with wastewater treatment discharges or sewage sludges. The presence of microplastics in wastewaters used for irrigation highlights the urgency of analysing the possible uptake of microplastics by crop plants. This study shows that submicrometre and micrometre plastic particles from treated wastewater enter the steles of crop plants via a crack entry at sites of lateral root emergence.

Production of the Plant Hormone Auxin by Salmonella and Its Role in the Interactions with Plants and Animals.

The ability of human enteric pathogens to colonize plants and use them as alternate hosts is now well established. Salmonella, similarly to phytobacteria, appears to be capable of producing the plant hormone auxin via an indole-3-pyruvate decarboxylase (IpdC), a key enzyme of the IPyA pathway. A deletion of the Salmonella ipdC significantly reduced auxin synthesis in laboratory culture. The Salmonella ipdC gene was expressed on root surfaces of Medicago truncatula. M. truncatula auxin-responsive GH3::GUS reporter was activated by the wild type Salmonella, and not but the ipdC mutant, implying that the bacterially produced IAA (Indole Acetic Acid) was detected by the seedlings. Seedling infections with the wild type Salmonella caused an increase in secondary root formation, which was not observed in the ipdC mutant. The wild type Salmonella cells were detected as aggregates at the sites of lateral root emergence, whereas the ipdC mutant cells were evenly distributed in the rhizosphere. However, both strains appeared to colonize seedlings well in growth pouch experiments. The ipdC mutant was also less virulent in a murine model of infection. When mice were infected by oral gavage, the ipdC mutant was as proficient as the wild type strain in colonization of the intestine, but it was defective in the ability to cross the intestinal barrier. Fewer cells of the ipdC mutant, compared with the wild type strain, were detected in Peyer's patches, spleen and in the liver. Orthologs of ipdC are found in all Salmonella genomes and are distributed among many animal pathogens and plant-associated bacteria of the Enterobacteriaceae, suggesting a broad ecological role of the IpdC-catalyzed pathway.

Role of LOTR1 in Nutrient Transport through Organization of Spatial Distribution of Root Endodermal Barriers

The formation of Casparian strips and suberin lamellae at the endodermis limits the free diffusion of nutrients and harmful substances via the apoplastic space between the soil solution and the stele in roots [1-3]. Casparian strips are ring-like lignin polymers deposited in the middle of anticlinal cell walls between endodermal cells and fill the gap between them [4-6]. Suberin lamellae are glycerolipid polymers covering the endodermal cells and likely function as a barrier to limit transmembrane movement of apoplastic solutes into the endodermal cells [7, 8]. However, the current knowledge on the formation of these two distinct endodermal barriers and their regulatory role in nutrient transport is still limited. Here, we identify an uncharacterized gene, LOTR1, essential for Casparian strip formation in Arabidopsis thaliana. The lotr1 mutants display altered localization of CASP1, an essential protein for Casparian strip formation [9], disrupted Casparian strips, ectopic suberization of endodermal cells, and low accumulation of shoot calcium (Ca). Degradation by expression of a suberin-degrading enzyme in the mutants revealed that the ectopic suberization at the endodermal cells limits Ca transport through the transmembrane pathway, thereby causing reduced Ca delivery to the shoot. Moreover, analysis of the mutants showed that suberin lamellae function as an apoplastic diffusion barrier to the steleat sites of lateral root emergence where Casparian strips are disrupted. Our findings suggest that thetransmembrane pathway through unsuberized endodermal cells, rather than the sites of lateral root emergence, mediates the transport of apoplastic substances such as Ca into the xylem.

Colonization and plant growth-promotion of tomato by Burkholderia tropica

Several diazotrophic Burkholderia species have been described to exhibit some activities involved in plant growth promotion and biological control. In this work seedlings of tomato plants were inoculated with this bacterium in order to study colonization of different vegetal tissues and plant growth promoting ability under greenhouse conditions. Tomato seedlings inoculated with Burkholderia tropica strain MTo-293 and two derivative strains containing the marker genes gusA and gfp, respectively (constructions described in this work), were grown under gnotobiotic conditions. Colonization was monitored both by colony counting of bacterial suspensions from homogenized tissues with or without previous surface disinfection and by microscopic observation of entire plant tissues. In another set of experiments tomato seedlings were inoculated with B. tropica MTo-293 for evaluation of tomato production under greenhouse conditions. Tomato yields were determined by quantifying total tomato production throughout the crop in two different seasons. B. tropica could be isolated from root surfaces (>7.0 logCFUg−1 fresh weight) and from surface-disinfected and disrupted roots (>5.0 logCFUg−1 fresh weight) and stems (>4.0 logCFUg−1 fresh weight) of inoculated plants. Microscopic studies showed colonizing bacteria on root hairs, root tips, lateral root emergence sites, and stomata. In greenhouse experiments inoculated plants showed a consistent increase of both number and weight of fruits as compared to uninoculated controls. Although this enhancement in fruit production was only statistically significant for fruit weight in the first crop season, our results show a consistent tendency to a higher yield (5–15%) for the inoculated treatments also in the second year. These results show that seedling inoculation with B. tropica led to effective root colonization of tomato plants followed by bacterial spreading to aerial tissues. This significant colonization was accompanied by an enhancement of tomato production in two different crop seasons.

Prediction of humic acids bioactivity using spectroscopy and multivariate analysis

Humic substances (HS) represent the organic material most widespread in nature and have positive effects on plant physiology influencing nutrient uptake and root architecture. Understanding the structure–activity relationship has been considered an essential step to drive technological approaches to produce HS with enhanced plant growth promotion performance. In this study we used solid state nuclear magnetic resonance (CP-MAS 13C NMR) and infrared spectroscopy with diffuse reflectance (DRIFT), a collection of data from humic acid-like substances (HA) obtained from vermicompost produced with different sources of raw material and maturation stage. The bioactivity was evaluated using the ability of HA to induce lateral root emergence sites as well acidification of the liquid medium by treated-maize root seedlings. The structure–activity relationship was done using multivariate analysis including principal component analysis (PCA), principal component regression (PCR) and Spearman correlation joined with NMR and DRIFT results. The main variables (higher loadings) correlated with HA bioactivity were for NMR positive loadings for lignin (56ppm, 124ppm, 148 and 153ppm) and COOH groups (174ppm) and negative loadings for carbohydrates (64, 75 and 102ppm). For DRIFT the main positive loadings were aryl stretching and bending (1560, 1480, 860 and 780cm−1) probably from lignins; for 2926cm−1 and 2852cm−1 due to CH asymmetric and symmetric stretching, respectively, probably from long chain fat acids due to the presence of rocking absorption band of (CH2)n with n>4 at 720cm−1 and negative loadings due to carbohydrate moieties (around 1100cm−1, CO stretching) and carboxylic acids (1724cm−1, CO stretching and 1220cm−1, COH stretching and COH bending). The use of spectroscopy data to attempt to relate structure to bioactivity may be of practical interest in developing plant growth promoters using renewed source of humic materials.

Monitoring the colonization of sugarcane and rice plants by the endophytic diazotrophic bacterium Gluconacetobacter diazotrophicus marked with gfp and gusA reporter genes

To evaluate the colonization process of sugarcane plantlets and hydroponically grown rice seedlings by Gluconacetobacter diazotrophicus strain PAL5 marked with the gusA and gfp reporter genes. Sugarcane plantlets inoculated in vitro with PAL5 carrying the gfp::gusA plasmid pHRGFPGUS did not present green fluorescence, but beta-glucuronidase (GUS)-stained bacteria could be observed inside sugarcane roots. To complement this existing inoculation methodology for micropropagated sugarcane with a more rapid colonization assay, we employed hydroponically grown gnotobiotic rice seedlings to study PAL5-plant interaction. PAL5 could be isolated from the root surface (10(8) CFU g(-1)) and from surface-disinfected root and stem tissues (10(4) CFU g(-1)) of inoculated plants, suggesting that PAL5 colonized the internal plant tissues. Light microscopy confirmed the presence of bacteria inside the root tissue. After inoculation of rice plantlets with PAL5 marked with the gfp plasmid pHRGFPTC, bright green fluorescent bacteria could be seen colonizing the rice root surface, mainly at the sites of lateral root emergence, at root caps and on root hairs. The plasmids pHRGFPGUS and pHRGFPTC are valid tools to mark PAL5 and monitor the colonization of micropropagated sugarcane and hydroponic rice seedlings. These tools are of use to: (i) study PAL5 mutants affected in bacteria-plant interactions, (ii) monitor plant colonization in real time and (iii) distinguish PAL5 from other bacteria during the study of mixed inoculants.

Transcriptional regulation of the cinnamyl alcohol dehydrogenase gene from sweetpotato in response to plant developmental stage and environmental stress

Cinnamyl alcohol dehydrogenase (CAD) is a key enzyme in the biosynthesis of lignin. We have isolated full length of a cDNA encoding CAD (IbCAD1) that was previously identified as the most abundant gene in an EST library of sweetpotato suspension cells. Phylogenetic analysis revealed that IbCAD1 belongs to the family of defense-related CADs. High levels of IbCAD1 mRNA were found in the roots of sweetpotato, but not in the leaves and petioles. The IbCAD1 gene transcripts were highly induced by cold, wounding, and reactive oxygen species. Analyses of transcriptional regulation of the IbCAD1 gene in transgenic tobacco plants carrying the IbCAD1 promoter–GUS revealed that IbCAD1 promoter expression was strong in the roots, but barely detectable in the cotyledons. IbCAD1 promoter activity increased with increasing root age, and strong promoter expression was observed in the lateral root emergence sites and in root tips. Weak GUS expression was observed in lignified tissues of vascular system of mature leaves and stems. IbCAD1 promoter activity was strongly induced in response to the biotic and abiotic stresses, with the strongest inducer being wounding, and was also induced by salicylic acid (SA) and jasmonic acid (JA) as well as by abscisic acid (ABA) and 6-benzylaminopurine. Taken together, our data suggest that IbCAD1 can be involved in JA- and SA-mediated wounding response and ABA-mediated cold response, respectively. The IbCAD1 gene may play a role in the resistance mechanism to biotic and abiotic stresses as well as in tissue-specific developmental lignification.

Studies on sodium bypass flow in lateral rootless mutants lrt1 and lrt2, and crown rootless mutant crl1 of rice (Oryza sativa L.)

An apoplastic pathway, the so-called bypass flow, is important for Na+ uptake in rice (Oryza sativa L.) under saline conditions; however, the precise site of entry is not yet known. We report the results of our test of the hypothesis that bypass flow of Na+ in rice occurs at the site where lateral roots emerge from the main roots. We investigated Na+ uptake and bypass flow in lateral rootless mutants (lrt1, lrt2), a crown rootless mutant (crl1), their wild types (Oochikara, Nipponbare and Taichung 65, respectively) and in seedlings of rice cv. IR36. The results showed that shoot Na+ concentration in lrt1, lrt2 and crl1 was lower (by 20-23%) than that of their wild types. In contrast, the bypass flow quantified using trisodium-8-hydroxy-1,3,6-pyrenetrisulphonic acid (PTS) was significantly increased in the mutants, from an average of 1.1% in the wild types to 3.2% in the mutants. Similarly, bypass flow in shoots of IR36 where the number of lateral and crown roots had been reduced through physical and hormonal manipulations was dramatically increased (from 5.6 to 12.5%) as compared to the controls. The results suggest that the path of bypass flow in rice is not at the sites of lateral root emergence.

Pathways of infection of Brassica napus roots by Leptosphaeria maculans

Infection of Brassica napus cotyledons and leaves by germinating ascospores of Leptosphaeria maculans leads to production of leaf lesions followed by stem cankers (blackleg). Leptosphaeria maculans also causes root rot but the pathway of infection has not been described. An L. maculans isolate expressing green fluorescent protein (GFP) was applied to the petiole of B. napus plants. Hyphal growth was followed by fluorescence microscopy and by culturing of sections of plant tissue on growth media. Leptosphaeria maculans grew within stem and hypocotyl tissue during the vegetative stages of plant growth, and proliferated into the roots within xylem vessels at the onset of flowering. Hyphae grew in all tissues in the stem and hypocotyl, but were restricted mainly to xylem tissue in the root. Leptosphaeria maculans also infected intact roots when inoculum was applied directly to them and hyphae entered at sites of lateral root emergence. Hyphal entry may occur at other sites but the mechanism is uncertain as penetration structures were not observed. Infection of B. napus roots by L. maculans can occur via above- and below-ground sources of inoculum, but the relative importance of the infection pathways under field conditions is unknown.

Endophytic Colonization of Vitis vinifera L. by Plant Growth-Promoting Bacterium Burkholderia sp. Strain PsJN

Patterns of colonization of Vitis vinifera L. cv. Chardonnay plantlets by a plant growth-promoting bacterium, Burkholderia sp. strain PsJN, were studied under gnotobiotic conditions. Wild-type strain PsJN and genetically engineered derivatives of this strain tagged with gfp (PsJN::gfp2x) or gusA (PsJN::gusA11) genes were used to enumerate and visualize tissue colonization. The rhizospheres of 4- to 5-week-old plantlets with five developed leaves were inoculated with bacterial suspensions. Epiphytic and endophytic colonization patterns were then monitored by dilution plating assays and microscopic observation of organ sections. Bacteria were chronologically detected first on root surfaces, then in root internal tissues, and finally in the fifth internode and the tissues of the fifth leaf. Analysis of the PsJN colonization patterns showed that this strain colonizes grapevine root surfaces, as well as cell walls and the whole surface of some rhizodermal cells. Cells were also abundant at lateral root emergence sites and root tips. Furthermore, cell wall-degrading endoglucanase and endopolygalacturonase secreted by PsJN explained how the bacterium gains entry into root internal tissues. Host defense reactions were observed in the exodermis and in several cortical cell layers. Bacteria were not observed on stem and leaf surfaces but were found in xylem vessels of the fifth internode and the fifth leaf of plantlets. Moreover, bacteria were more abundant in the fifth leaf than in the fifth internode and were found in substomatal chambers. Thus, it seems that Burkholderia sp. strain PsJN induces a local host defense reaction and systemically spreads to aerial parts through the transpiration stream.

Combined use of different Gfp reporters for monitoring single-cell activities of a genetically modified PCB degrader in the rhizosphere of alfalfa

Single-cell localization and activity of Pseudomonas fluorescens F113, colonizing alfalfa roots, were monitored using fusions of the Escherichia coli rrnBP1 ribosomal promoter and gfp genes encoding green fluorescent protein (Gfp) of different stability. The monitoring systems permitted non-destructive in situ detection of F113rifpcb cells on the entire root system grown in both the presence and absence of 3-chlorobiphenyl (PCB-2). The root tip and sites of lateral root emergence were found to be hotspots for fast-growing cells. In addition, a reporter strain of P. fluorescens F113rifpcb for degradation of chlorinated biphenyl was constructed, using another gfp fusion with the meta-pathway Pm promoter from Pseudomonas putida (TOL plasmid). Expression of this promoter, which is strongly induced by the PCB-2 degradation product, 3-chlorobenzoate, was tested in vitro and subsequently monitored in vivo on alfalfa roots using the P. fluorescens F113rifpcb reporter. A small but distinct fraction of the introduced bacteria activated the Pm promoter and thus appeared to sense a PCB-2 degradation product in the alfalfa rhizosphere. The degrading cells, which by design were identical to the sensing cells, were located in distinct microcolonies on the root surface or in intercellular crevices between the root epidermal cells. However, PCB-degrading cells were not observed in the root areas containing fast-growing cells, indicating that PCB degradation was not linked to high cellular activity.

Humic acids isolated from earthworm compost enhance root elongation, lateral root emergence, and plasma membrane H+-ATPase activity in maize roots.

Earthworms (Eisenia foetida) produce humic substances that can influence plant growth by mechanisms that are not yet clear. In this work, we investigated the effects of humic acids (HAs) isolated from cattle manure earthworm compost on the earliest stages of lateral root development and on the plasma membrane H(+)-ATPase activity. These HAs enhance the root growth of maize (Zea mays) seedlings in conjunction with a marked proliferation of sites of lateral root emergence. They also stimulate the plasma membrane H(+)-ATPase activity, apparently associated with an ability to promote expression of this enzyme. In addition, structural analysis reveals the presence of exchangeable auxin groups in the macrostructure of the earthworm compost HA. These results may shed light on the hormonal activity that has been postulated for these humic substances.

Surface colonization of lodgepole pine (Pinus contorta var. latifolia [Dougl. engelm.]) roots by Pseudomonas fluorescens and Paenibacillus polymyxa under gnotobiotic conditions

Indirect immunofluorescence techniques and confocal scanning laser microscopy were used to identify rhizobacterial strains on the root surfaces of pine seedlings, which were grown from seeds under gnotobiotic conditions. Conifer plant growth promoting rhizobacterial strains Paenibacillus polymyxa L6 and Pw-2, and the forest soil isolate Pseudomonas fluorescens M20, were inoculated onto surface-disinfested pine seeds, singly, or in dual combinations: strains L6 + M20, or strains Pw-2 + M20. Segments containing particular root microsites (root tip, root hair zone, or areas of lateral root emergence) were sampled randomly from roots 7 or 13 weeks after inoculation, and the colonization of roots by each bacterium was observed. Root segments were also sampled from individual roots at six different points along the length of the root, and the qualitative colonization of younger areas, closer to the root tip, contrasted with that of older areas, closer to the root base. The ability of strain M20 to colonize root areas adjacent to sites of lateral root emergence improves in the presence of either P. polymyxa strain, while the ability of the P. polymyxa strains to colonize these areas was not affected. More rhizobacteria were also generally observed on younger root tissues than on areas closer to the root base.

Soil and plant specific effects on bacterial community composition in the rhizosphere

Eubacterial community structures in the plant rhizosphere were examined with respect to plant species, soil type, and root zone location. Three plant species (chickpea, rape and Sudan grass) were grown in intact cores of three California soils (a sandy soil, a sandy loam, and a clay) and were provided with a complete fertilizer solution with or without nitrogen supplied as ammonium nitrate. After 7.5 weeks, the plants were harvested and DNA was extracted from soil adhering to the root tips and from mature root zones at the sites of lateral root emergence. Eubacterial community structures were examined by PCR-DGGE of 16S rDNA to determine the relative abundance and species diversity. While both soil type and nitrogen fertilization affected plant growth, canonical correspondence analyses showed that nitrogen had no significant effect on eubacterial community structures. Eubacterial species diversity was higher in the mature root zones than at the root tips in the sandy soil and the clay but not in the loamy sand. Monte Carlo permutation tests indicated that plant species, root zone and soil type as well as the interactions between these variables had significant effects on community structure. The bacterial rhizosphere community of chickpea was influenced primarily by soil type, whereas root zone was less important. In contrast to chickpea, the community in the rhizosphere of rape and Sudan grass was more affected by the root zone than the soil type. In the sandy soil and the loamy sand, the eubacterial rhizosphere community structure was more affected by the root zone than the plant species and the three plant species had distinct communities. In the clay however, the root zone was less important than the plant species and the rhizosphere communities of chickpea differed from those of rape and Sudan grass. It is concluded that the bacterial community composition in the rhizosphere is affected by a complex interaction between soil type, plant species and root zone location.

Rhizobia can induce nodules in white clover by "hijacking" mature cortical cells activated during lateral root development.

We examined a range of responses of root cortical cells to Rhizobium sp. inoculation to investigate why rhizobia preferentially nodulate legume roots in the zone of emerging root hairs, but generally fail to nodulate the mature root. We tested whether the inability to form nodules in the mature root is due to a lack of plant flavonoids to induce the bacterial genes required for nodulation or a failure of mature cortical cells to respond to Rhizobium spp. When rhizobia were inoculated in the zone of emerging root hairs, changes in beta-glucuronidase (GUS) expression from an auxin-responsive promoter (GH3), expression from three chalcone synthase promoters, and the accumulation of specific flavonoid compounds occurred in cortical cells prior to nodule formation. Rhizobia failed to induce these responses when inoculated in the mature root, even when co-inoculated with nod gene-inducing flavonoids. However, mature root hairs remained responsive to rhizobia and could support infection thread formation. This suggests that a deficiency in signal transduction is the reason for nodulation failure in the mature root. However, nodules could be initiated in the mature root at sites of lateral root emergence. A comparison between lateral root and nodule formation showed that similar patterns of GH3:gusA expression, chalcone synthase gene expression, and accumulation of a particular flavonoid compound occurred in the cortical cells involved in both processes. The results suggest that rhizobia can "hijack" cortical cells next to lateral root emergence sites because some of the early responses required for nodule formation have already been activated by the plant in those cells.