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[Morphology of wheat roots under low-phosphorus stress].

The morphology of root systems of different wheat (Triticum aestivum L.) genotypes under low-phosphorus stress were studied to determine the effects of external factors on components of root system and the early morphological indicators related to phosphorus efficiency. The number of root axes and the length of lateral root of P-deficient plant were significantly lower than those of P-sufficient plant. The length of root axis and root system, and the number of lateral roots were sharply increased under low-P stress. The number and length of root axis were significantly different under different levels of phosphorus supply and among different wheat genotypes under same phosphorus supply. This implied that the two traits (number and length of root axis) were controlled by genotype and external factors. The difference in the characteristics of lateral root of the given wheat genotypes was significant only between different levels of P supply. It showed that the traits of lateral root mainly depended on external factors. The length and number of root axis, root length, and root angle were significantly different among 6 wheat genotypes. There exited significant linear relationships between relative grain yield and the interaction of the morphological traits, and it implied that the traits could be used as early indicators of selecting high P-efficiency wheat varieties.

Minerals↗

Root stimulated nitrogen removal: only a local effect or important for water treatment?

Plants in constructed wetlands serve as carriers for attached microbial growth. They mainly transfer oxygen and release exsudates to the root zone. In consequence of this an area around the roots, called the rhizosphere exists, in which bacteria are stimulated by root growth. The goals were to ascertain whether stimulating the microbial cenosis only has a local effect on the rhizoplane, and to establish the importance of this stimulation for the water purification process in the root zone. Observations were carried out in a laboratory batch reactor filled with sand and planted with reeds (Phragmites australis). A small section was separated with gauze to avoid root growth outside this zone. The reactor was incubated with an artificial waste water containing a high concentration of ammonium. Samples were taken at intervals of 10 mm away from the gauze. The chemical and physical conditions and enzyme activities in soil sections at different distances from the roots affecting the efficiency of nitrogen removal were characterized. An influence was detectable by several parameters up to a specific root distance. Indirect parameters such as the total bacterial number and the DNA amount seem to be affected up to a distance of 50 mm from the root whereas the oxygen amount and DOC are unaffected at a distance exceeding 20-30 mm. This is an initial indication that improved nitrogen removal is also possible in the wider root surroundings. In view of the average root-to-root distance of 35 mm, the root-influenced area could therefore be expanded to the whole rooted zone in a constructed wetland. The influence on bacteria by roots is not just a local effect but may also play an important role in the whole purification process.

Biodegradation, Environmental↗

Radiographic evaluation of periapical healing after obturation of infected root canals: an in vivo study.

AIM: To radiographically compare periapical repair of roots with infected root canals obturated in one-step or with calcium hydroxide (Ca(OH)2) intracanal medication in two steps. METHODOLOGY: Standardized preoperative periapical radiographs were taken of 72 roots of vital dogs' teeth. All roots were then aseptically instrumented to ISO size 45. As negative controls, 12 roots were aseptically obturated. The remaining roots were infected with dental plaque and closed. Six weeks later, apical periodontitis was radiographically confirmed in the infected roots. The roots were divided into the following groups: group 1, one-step (n = 24); roots were irrigated with 10 cc saline, obturated, and permanently restored. Group 2, Ca(OH)2 (n = 24); roots were treated as in group 1, except that after saline irrigation Ca(OH)2 medicament was placed in the canal 1 week before obturation. Group 3, positive control (n = 12); the roots were irrigated with saline, access permanently closed but canals not obturated. Group 4, negative control (n = 12); previously aseptically obturated roots were permanently restored. After 6 months, standardized postoperative radiographs were obtained. Three independent evaluators blinded to the treatment groups evaluated the preoperative and postoperative radiographs. The evaluators were instructed to rate each root, based on changes on the radiographs, as failed, improved or healed. RESULTS: Radiographically, the percentage of cases that completely healed were similar for the one-step and Ca(OH)2 groups (35.3% vs. 36.8%). However, the Ca(OH)2 group had fewer failed cases (15.8% vs. 41.2%) and more improved cases (47.4% vs. 23.5%) than the one-step group. CONCLUSION: Power statistics demonstrated that at 43 cases per group, Ca(OH)2 treatment would be statistically superior to one-step treatment. We consider this number to be clinically important.

Animals↗

Induction and growth properties of carrot roots with different complements of Agrobacterium rhizogenes T-DNA.

Single and multiple infections of carrot discs were carried out with Agrobacterium strains harbouring different segments of pRi1855 TL-DNA cloned in the binary vector Bin 19 and with a strain carrying the TR-DNA from the same Ri plasmid. Roots induced by the various co-inoculations were cultured and their growth patterns were followed. Abundant roots could be induced by TL-DNA rol genes A, B and C as a single insert (rolA + B + C) and by rolB alone provided an extended segment beyond its 5' non-coding region was included in the construction. A depression of rooting capability was caused by the inclusion of rolC together with rolB (rolB + C). In all cases co-inoculation with the Agrobacterium carrying TR-DNA-borne auxin genes was necessary for root induction since none of the rol constructions was in itself capable of eliciting any response; an exceeding majority of these roots were however shown to contain rol genes but no TR-DNA. Rooting was also elicited if rol constructions were co-inoculated with a strain carrying TL-DNA genes 13 and 14 (ORF13 + 14) instead of the TR-DNA strain. These roots were shown to contain both rol genes and ORF13 + 14. Striking differences in growth properties were shown by roots containing different complements of TL-DNA genes. Typical hairy root traits, high growth rate, branching and, most noticeably, absence of geotropism, were shown by roots containing rolB alone, while roots with rolA + B + C were geotropic as normal carrot roots. Hairy root traits were conferred to rolA + B + C roots by the concomitant presence of ORF13 + 14 and by the addition of auxin to the culture medium. A model is presented which attempts to rationalize the growth patterns by assigning interplaying roles to the various TL-DNA genes involved.

Blotting, Southern↗

Moving Waves of Bacterial Populations and Total Organic Carbon along Roots of Wheat.

> Abstract To determine if spatial variation in soluble carbon sources along the root coincides with different trophic groups of bacteria, copiotrophic and oligotrophic bacteria were enumerated from bulk soil and rhizosphere samples at 2 cm intervals along wheat roots 2, 3, and 4 weeks after planting. There was a moderate rhizosphere effect in one experiment with soil rich in fresh plant debris, and a very pronounced rhizosphere effect in the second experiment with soil low in organic matter. We obtained wavelike patterns of both trophic groups of bacteria as well as water-soluble total organic carbon (TOC) along the whole root length (60 or 90 cm). TOC concentrations were maximal at the root tip and base and minimal in the middle part of the roots. Oscillations in populations of copiotrophic and oligotrophic bacteria had two maxima close to the root tip and at the root base, or three maxima close to the tip, in the middle section, and at the root base. The location and pattern of the waves in bacterial populations changed progressively from week to week and was not consistently correlated with TOC concentrations or the location of lateral root formation. Thus, the traditional view that patterns in bacterial numbers along the root directly reflect patterns in exudation and rhizodeposition from several fixed sources along the root may not be true. We attributed the observed wavelike patterns in bacterial populations to bacterial growth and death cycles (due to autolysis or grazing by predators). Considering the root tip as a moving nutrient source, temporal oscillations in bacterial populations at any location where the root tip passed would result in moving waves along the root. This change in concept about bacterial populations in the rhizosphere could have significant implications for plant growth promotion and bioremediation.

Journal Article↗

Age related sprouting of dorsal root axons after sensory denervation.

To test for sprouting of dorsal root axons, 3 consecutive dorsal roots were sectioned above and 3 were sectioned below a spared root in rats 1 month and 24 months old. At various survival times, myelinated and unmyelinated axons were counted in the spared root, the first cranial root and the first caudal root, and these counts were compared to similar counts from the same roots on the unoperated side. The counts of myelinated axons in all roots and of unmyelinated axons in the caudal roots were not different from the counts in the same roots on the unoperated side. Thus there is no evidence for sprouting of these axons in our system. By contrast, the unmyelinated axons in the cranial and spared roots were significantly increased as compared to the unmyelinated axons in the same roots on the unoperated side. We interpret this increase to be sprouting of dorsal root axons. To determine the effects of age, animals operated at 1 month and sacrificed 1 month, 3 months or 18 months later were compared with animals operated at 2 years and sacrificed 1 month later. The unmyelinated axons were greater by approximately 20% on the operated side in the cranial and spared roots in the young rats and by approximately 8% in old rats. Thus the sprouting of unmyelinated axons is more vigorous when the surgery is performed in young animals, but there still seems to be a response in older animals.

Aging↗

Aortic root replacement--20 years experience of the use of homografts.

The aortic root was replaced with a free root homograft 100 times in 91 patients from 1970 to January 1987. The ages of the patients were in the range 5-73 years (mean 29.7). There were 63 males and 34 females. Thirty had endocarditis with root abscesses. Forty-one had congenitally small aortic roots and 20 patients had such severe root calcification that root replacement was required. One patient had an aortic sinus aneurysm repaired using this technique. Operative mortality (overall 24%) analysed by operative indication was highest for prosthetic endocarditis (33%). 20% of those with root calcification and 15% of those operated on for hypoplastic roots died. Endocarditis was cured in 19 of 20 survivors. One patient needed repeat root replacement to achieve this. Thirteen patients out of 35 survivors in the hypoplastic group were restudied. Mean subvalvular gradient was 12 +/- 9 mmHg (range 0-27 mmHg). Late mortality was lowest in the endocarditis group (5%), and 11% in the hypoplastic group. There were 4 (24%) late deaths in the root calcification group but 2 were non cardiac. Eight homograft roots failed, two in the endocarditis group and one in the hypoplastic group due to endocarditis. The remainder failed because of 'wear and tear'. Five were replaced using homografts and one by a composite graft. Two were treated by prosthetic subcoronary aortic valve replacement with pericardial patch enlargement of the root. A further patient with Marfan's syndrome had an infected composite graft. This was replaced by a homograft root. There are 5 long term survivors of 7 repeat aortic root replacements.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Rooting phylogenetic trees with distant outgroups: a case study from the commelinoid monocots.

Phylogenetic rooting experiments demonstrate that two chloroplast genes from commelinoid monocot taxa that represent the closest living relatives of the pickerelweed family, Pontederiaceae, retain measurable signals regarding the position of that family's root. The rooting preferences of the chloroplast sequences were compared with those for artificial sequences that correspond to outgroups so divergent that their signal has been lost completely. These random sequences prefer the three longest branches in the unrooted ingroup topology and do not preferentially root on the branches favored by real outgroup sequences. However, the rooting behavior of the artificial sequences is not a simple function of branch length. The random outgroups preferentially root on long terminal ingroup branches, but many ingroup branches comparable in length to those favored by random sequences attract no or few hits. Nonterminal ingroup branches are generally avoided, regardless of their length. Comparisons of the ease of forcing sequences onto suboptimal roots indicate that real outgroups require a substantially greater rooting penalty than random outgroups for around half of the least-parsimonious candidate roots. Although this supports the existence of nonrandomized signal in the real outgroups, it also indicates that there is little power to choose among the optimal and nearly optimal rooting possibilities. A likelihood-based test rejects the hypothesis that all rootings of the subtree using real outgroup sequences are equally good explanations of the data and also eliminates around half of the least optimal candidate roots. Adding genes or outgroups can improve the ability to discriminate among different root locations. Rooting discriminatory power is shown to be stronger, in general, for more closely related outgroups and is highly correlated among different real outgroups, genes, and optimality criteria.

DNA, Chloroplast↗

Growth, Gravitropism, and Endogenous Ion Currents of Cress Roots (Lepidium sativum L.) : Measurements Using a Novel Three-Dimensional Recording Probe.

A novel, three-dimensional recording, vibrating probe was used for measuring the density and direction of the endogenous ionic current of cress roots (Lepidium sativum L.) bathed in low salt media (artificial pond water, APW). Roots submerged in regular APW and growing vertically show the following current pattern. Current of 0.7 microampere/square centimeter density enters or leaves the root cap; the current changes direction frequently. Current of 1.6 microamperes/square centimeter enters the meristem zone most of the time. Maximum current with a density of 2.2 microamperes/square centimeter enters the apical elongating zone, i.e. between 0.8 and 1.2 millimeters behind the root tip. The current density decreases to 1.4 microamperes/square centimeter at 2 millimeters, i.e. in the central elongating zone, and to 1.0 microampere/square centimeter at 3 millimeters, i.e. in the basal elongating zone. The current direction changes from inward to predominantly outward between 1.2 and 3 millimeters behind the tip. Measurements on opposite flanks of the roots indicate that the current pattern is fairly symmetrical. After placing the roots horizontally, the density of the endogenous current remains stable, but the current direction changes at the root cap and in the meristem zone. The current leaves the root on the upper side and enters on the lower side, causing a highly asymmetrical current pattern at the very tip. The current pattern at the upper and lower side further away from the tip remains the same as in vertical roots. Roots submerged in low Ca(2+) APW show a very different current pattern, no gravitropism, and no change of the current pattern after horizontal orientation. In these roots current enters the root cap and the basal elongating zone and leaves the apical elongating zone. Three conclusions are drawn from these results: First, plant roots elongate by two different modes of growth that are correlated with different current directions. They grow by cytoplasmic enlargement at sites of inward current and by turgor-driven elongation at sites of outward current. Second, a change in the current pattern at the root cap and in the meristem zone is a clear indicator of later gravitropism. Third, Ca(2+) ions are involved in the gravistimulated change in the current pattern, probably affecting the activity of plasmalemma H(+)-ATPases.

Journal Article↗

Phosphorus Stress-Induced Proteoid Roots Show Altered Metabolism in Lupinus albus.

Proteoid roots develop in Lupinus albus L. in response to nutrient stress, especially P. Proteoid roots excrete citrate and thus increase the availability of P, Fe, and Mn in the rhizosphere. In an effort to understand citrate synthesis and organic acid metabolism in proteoid roots of lupin, we have evaluated in vitro enzyme activities of citrate synthase (CS), malate dehydrogenase (MDH), and phosphoenolpyruvate carboxylase (PEPC) in proteoid and normal roots of plants grown with or without P. Organic acid concentrations, respiration rates, and dark 14CO2-labeling patterns were also determined. The in vitro specific activities of CS, MDH, and PEPC and in vivo dark 14CO2 fixation were higher in proteoid roots compared to normal roots, particularly under P stress. Western blot analysis showed that PEPC enzyme protein was more highly expressed in -P proteoid roots compared to other tissues. The majority of the fixed 14C was found in organic acids, predominantly malate and citrate. A larger fraction of citrate was labeled in P- stressed proteoid roots compared to other root tissue. Respiration rates of proteoid roots were 31% less than those of normal roots. The data provide evidence for increased synthesis of citrate in proteoid roots compared to normal roots, particularly under P stress. A portion of the carbon for citrate synthesis is derived from nonautotrophic CO2 fixation via PEPC in proteoid roots.

Journal Article↗

Root growth and oxygen relations at low water potentials. Impact Of oxygen availability in polyethylene glycol solutions

Polyethylene glycol (PEG), which is often used to impose low water potentials (psiw) in solution culture, decreases O2 movement by increasing solution viscosity. We investigated whether this property causes O2 deficiency that affects the elongation or metabolism of maize (Zea mays L.) primary roots. Seedlings grown in vigorously aerated PEG solutions at ambient solution O2 partial pressure (pO2) had decreased steady-state root elongation rates, increased root-tip alanine concentrations, and decreased root-tip proline concentrations relative to seedlings grown in PEG solutions of above-ambient pO2 (alanine and proline accumulation are responses to hypoxia and low psiw, respectively). Measurements of root pO2 were made using an O2 microsensor to ensure that increased solution pO2 did not increase root pO2 above physiological levels. In oxygenated PEG solutions that gave maximal root elongation rates, root pO2 was similar to or less than (depending on depth in the tissue) pO2 of roots growing in vermiculite at the same psiw. Even without PEG, high solution pO2 was necessary to raise root pO2 to the levels found in vermiculite-grown roots. Vermiculite was used for comparison because it has large air spaces that allow free movement of O2 to the root surface. The results show that supplemental oxygenation is required to avoid hypoxia in PEG solutions. Also, the data suggest that the O2 demand of the root elongation zone may be greater at low relative to high psiw, compounding the effect of PEG on O2 supply. Under O2-sufficient conditions root elongation was substantially less sensitive to the low psiw imposed by PEG than that imposed by dry vermiculite.

Journal Article↗

Early Events in the Infection of Soybean (Glycine max L. Merr) by Rhizobium japonicum: I. LOCALIZATION OF INFECTIBLE ROOT CELLS.

The infectible cells of soybean roots appear to be located at any given time just above the zone of root elongation and just below the position of the smallest emergent root hairs. The location of infectible cells on the primary root at the time of inoculation was inferred from the position of subsequent nodule development, correcting for displacement of epidermal cells due to root elongation. Marks were made on the seedling growth pouches at the time of inoculation to indicate the position of the root tip and the zones of root hair development. Virtually all of the seedlings developed nodules on the primary root above the marks made at the root tips at the time of inoculation. None of the plants formed nodules on the root where mature root hairs were present at the time of inoculation. These results and profiles of nodulation frequency indicate that the location of infectible cells is developmentally restricted. When inoculations were delayed for intervals of 1 to 4 hours after marking the positions of the root tips, progressively fewer nodules were formed above the root tip marks, and the uppermost of these nodules were formed at progressively shorter distances above the marks. These results indicate that the infectibility of given host cells is a transient property that appears and then is lost within a few hours. The results also indicate that host responses leading to infection and nodulation are triggered or initiated in less than 2 hours after inoculation. The extent of nodulation above the root tip mark increased in proportion to the logarithm of the number of bacteria in the inoculum.

Journal Article↗

Rapid Changes in the Pattern of Electric Current around the Root Tip of Lepidium sativum L. following Gravistimulation.

Using a highly sensitive vibrating electrode, the pattern of naturally occurring electric currents around 1-day-old primary roots of Lepidium sativum L. growing vertically downward and the current pattern following gravistimulation of the root has been examined. A more or less symmetrical pattern of current was found around vertically oriented, downward growing roots. Current entered the root at the root cap, the meristem, and the beginning of the elongation zone and left the root along most of the elongation zone and in the root hair zone. After the root was tilted to a horizontal position, we observed current flowing acropetally at the upper side of the root cap and basipetally at the lower side within about 30 seconds in most cases. After a delay of several minutes, acropetally oriented current was also found flowing along the upper side of the meristematic zone. The apparent density of the acropetal current in the root cap region increased and then decreased with time. Gravitropic curvature was first visible approximately 10 minutes after tilting of the root to the horizontal position. Since the change in the pattern of current in the root cap region precedes bending of the root and is different for the upper and lower side, a close connection is suggested between the current and the transduction of information from the root cap to the elongation zone following graviperception in the cap.

Journal Article↗

Abscisic Acid Accumulation by Roots of Xanthium strumarium L. and Lycopersicon esculentum Mill. in Relation to Water Stress.

Plants of Xanthium strumarium L. and Lycopersicon esculentum Mill. cv ;Rheinlands Ruhm' were grown in solution culture, and control and steam-girdled intact plants were stressed. Detached roots of both species were stressed to different extents in two ways: (a) either in warm air or, (b) in the osmoticum Aquacide III. The roots of both species produced and accumulated progressively more abscisic acid (ABA), the greater the stress inflicted by either method. ABA-glucose ester levels in Xanthium roots were not affected by water stress and were too low to be the source of the stress-induced ABA. The fact that ABA accumulated in detached roots and in roots of girdled plants proves that ABA was synthesized in the roots and not merely transported from the shoots.Maximum ABA accumulation in detached roots occurred after 60 to 70% loss of fresh weight. In Xanthium roots, ABA levels continued to increase for at least 11 hours, and no catabolism was apparent when stressed roots were immersed in water, although the roots did stop accumulating ABA. When osmotically stressed, Xanthium roots reached a maximum ABA level after 2 hours, but ABA continued to rise in the medium.Under optimal stress conditions, endogenous ABA levels increased 100 times over their prestress values in detached roots of Xanthium, and 15 times in Lycopersicon under nonoptimal stress, when endogenous ABA was expressed as concentrations based on tissue water content. These are much greater relative increases than observed in the leaves (15 times in Xanthium, 3 times in Lycopersicon), although the roots contain substantially less ABA than the leaves in all circumstances. The results suggest that the endogenous level of ABA in roots could rise appreciably prior to leaf wilt, and could modify the plant's water economy before the leaves become stressed.

Journal Article↗

Effect of localized nitrate application on isoflavonoid concentration and nodulation in split-root systems of wild-type and nodulation-mutant soybean plants.

Although isoflavonoids are known to be inducers of nod genes in Bradyrhizobium japonicum, it was recently proposed that internal root levels of isoflavonoids may be important in nodule development on soybean (Glycine max [L.] Merr.). The hypernodulating soybean mutants were shown to accumulate higher root concentrations of isoflavonoid compounds (daidzein, genistein, and coumestrol) and to be more extensively nodulated than was the Williams parent when inoculated with B. japonicum. The hypernodulating mutants and the parent line, Williams, also showed decreased isoflavonoid concentrations and decreased nodule development if N was applied. The current study evaluated the effect of localized NO(3) (-) application on root isoflavonoid concentration and on nodulation in split-root systems of the Williams wild type and a hypernodulating mutant (NOD1-3). Nitrate application markedly decreased isoflavonoid concentrations in non-inoculated soybean roots. When roots were inoculated, nodule number, weight, and nitrogenase activity were markedly suppressed on the root-half receiving 5 millimolar NO(3) (-) compared with the other root-half receiving 0 millimolar NO(3) (-). High performance liquid chromatographic analyses of root extracts showed that the root-half receiving 5 millimolar NO(3) (-) was markedly lower in isoflavonoid concentrations in both soybean lines. This was partially due to the localized stimulatory effect of NO(3) (-) on root growth. The inoculated NOD1-3 mutant had higher isoflavonoid concentrations than did the Williams control in both the presence and absence of NO(3) (-). These results provide evidence that the site of N application primarily controls the site of nodulation inhibition, possibly through decreasing isoflavonoid levels. Although the effect of NO(3) (-) on nodule development and root isoflavonoid concentration was strongly localized, there was evidence that NO(3) (-) also resulted in a systemic effect on root isoflavonoids. The results are consistent with previous speculation that internal levels of root isoflavonoids may affect nodule development.

Journal Article↗

Growth and Movement of Spot Inoculated Rhizobium meliloti on the Root Surface of Alfalfa.

Inoculum droplets of approximately 10 nanoliter volume and containing about 10 Rhizobium meliloti cells were placed onto the root surface of alfalfa seedlings in plastic growth pouches at either the root tip, the position of the smallest emergent root hairs, or at a site midway between these points. The droplets were initially confined to an area of about 0.2 square millimeter at the point of application. By 48 and 96 hours after inoculation, the inoculum bacteria and their progeny were distributed over several centimeters of the root between the initial site of deposition and the growing root tip, reaching densities of 10(3) to 10(4) bacteria per centimeter near the site of initial deposition and decreasing exponentially from that point toward the root tip. Graphite particles deposited on the root surface close to the growing tip were similarly distributed along the root length by 48 and 96 hours, suggesting that passive displacement by root cell elongation was primarily responsible for the spread of bacteria. A nonmotile mutant of R. meliloti colonized alfalfa roots to the same extent as the wild type and was usually distributed in the same manner, indicating that bacterial motility contributed little under these conditions to long distance spread of the bacteria. However, when applied in low numbers, R. meliloti mutants defective in motility or chemotaxis were considerably less efficient in initiating nodules near the point of inoculation than the wild type. This implies that motility and/or chemotaxis contribute significantly to local exploration for suitable infection sites. Almost all nodules on the primary root formed within a few millimeters of the spot-inoculation site, indicating that, under our experimental conditions, movement and multiplication of R. meliloti on the root surface were not sufficient to maintain an adequate population in the infectible region of the root during root growth.

Journal Article↗

Specific phases of root hair attachment in the Rhizobium trifolii-clover symbiosis.

The time course and orientation of attachment of Rhizobium trifolii 0403 to white clover root hairs was examined in slide cultures by light and electron microscopy. Inocula were grown for 5 days on defined BIII agar medium and represented the large subpopulation of fully encapsulated single cells which uniformly bind the clover lectin trifoliin A. When 10(7) cells or more were added per seedling, bacteria attached within minutes, forming randomly oriented clumps at the root hair tips. Several hours later, single cells attached polarly to the sides of the root hair. This sequence of attachment to clover root hairs was selective for R. trifolii at inoculum sizes of 10(7) to 4 X 10(8) per seedling, specifically inhibited if 2-deoxy-D-glucose, a hapten for trifoliin A, was present in the inoculum, and not observed when 4 X 10(8) cells were added to alfalfa seedling roots or to large clover root cell wall fragments which lacked trifoliin A but still had trifoliin A receptors. Once attached, R. trifolii 0403 became progressively less detachable with 2-deoxy-D-glucose. At smaller inoculum sizes (10(5) to 10(6) cells per seedling), there was no immediate clumping of R. trifolii at clover root hair tips, although polar binding of bacteria along the root hair surface was observed after 4 h. The interface between polarly attached bacteria and the root hair cell wall was shown to contain trifoliin A by immunofluorescence microscopy. Also, this interface was shown by transmission electron microscopy to contain electron-dense granules of host origin. Scanning electron microscopy revealed an accumulation of extracellular microfibrils associated with the lateral and polar surfaces of the attached bacteria, detectable after 12 h of incubation with seedling roots. At this same time, there was a significant reduction in the effectiveness of 2-deoxy-D-glucose in dislodging bacteria already attached to root hairs and an increase in firm attachment of bacteria to the root hair surface, which withstood the hydrodynamic shear forces of high-speed vortexing. These results are interpreted as a sequence of phases in attachment, beginning with specific reversible interactions between bacterial and plant surfaces (phase I attachment), followed by production of extracellular microfibrils which firmly anchor the bacterium to the root hair (phase 2 adhesion). Thus, attachment of R. trifolii to clover root hairs is a specific process requiring more than just the inherent adhesiveness of the bacteria to the plant cell wall.(ABSTRACT TRUNCATED AT 400 WORDS)

Microscopy, Phase-Contrast↗

Effects of fungal root pathogens on the population dynamics of biocontrol strains of fluorescent pseudomonads in the wheat rhizosphere.

The influences of Gaeumannomyces graminis var. tritici (which causes take-all of wheat), Rhizoctonia solani AG-8 (which causes rhizoctonia root rot of wheat), Pythium irregulare, P. aristosporum, and P. ultimum var. sporangiiferum (which cause pythium root rot of wheat) on the population dynamics of Pseudomonas fluorescens 2-79 and Q72a-80 (bicontrol strains active against take-all and pythium root rot of wheat, respectively) in the wheat rhizosphere were examined. Root infection by either G. graminis var. tritici or R. solani resulted in populations of both bacterial strains that were equal to or significantly larger than their respective populations maintained on roots in the absence of these pathogens. In contrast, the population of strain 2-79 was significantly smaller on roots in the presence of any of the three Pythium species than on noninfected roots and was often below the limits of detection (50 CFU/cm of root) on Pythium-infected roots after 40 days of plant growth. In the presence of either P. aristosporum or P. ultimum var. sporangiiferum, the decline in the population of Q72a-80 was similar to that observed on noninfected roots; however, the population of this strain declined more rapidly on roots infected by P. irregulare than on noninfected roots. Application of metalaxyl (which is selectively inhibitory to Pythium spp.) to soil naturally infestated with Pythium spp. resulted in significantly larger rhizosphere populations of the introduced bacteria over time than on plants grown in the same soil without metalaxyl. It is apparent that root infections by fungal pathogens may either enhance or depress the population of fluorescent pseudomonads introduced for their control, with different strains of pseudomonads reacting differentially to different genera and species of the root pathogens.

Journal Article↗