N2(C2H2−C2H4) fixation in two species of Ceanothus seedlings in second year postfire chaparral

Nitrogen fixation in excised root nodules of 2-year-old, postfireCeanothus tomentosus andC. leucodermis seedlings was measured over an 8-month period using the acetylene reduction method. High levels of NO₃–N and NH₄–N present in postfire soils were limited to the upper 10 cm and did not inhibit nodulation in these deeper-rooting seedlings. Decreases in acetylene reduction activity occurred with decreased soil moisture and increased soil temperature. Nitrogen gains from these two Ceanothus shrub seedlings totalled 1.6 kg N ha^–1 yr^–1.     

Soil and environmental factors related to nodulation in Cowania and Purshia

Summary Nitrogen fixing trees and shrubs may be useful in revegetation efforts. The possibility that soil and environmental factors may influence a soil's capability to produce nodulated seedlings was explored.Purshia tridentata andCowania mexicana var. Stansburiana seedlings were grown in greenhouse trials using ten soils from native sites for each of the two genera. Treatments included a control and a six mmole nitrogen amendment as NH₄NO₃ for both surface and subsurface samples. Nodulation was often sparse for seedlings grown in surface collected samples. Although nodulation was usually better in subsoil samples, even some subsoils produced few or no nodules. Nitrogen additions inhibit nodulation and although soil nitrogen may be inhibitory in some unamended surface soils it is probably not a general cause of sparse nodulation. Nodule masses showed the same trends as nodule number but varied less with treatment and depth of soil source. Seedlings compensated for sparse nodulation with an increase in mass per nodule. Incidence of nodulation was related to some soil and environmental factors. Multiple regression analysis explained a substantial portion of nodulation variability. Soils from lower elevations with less precipitation did not produce well nodulated seedlings even in well watered greenhouse trials. Micronutrient cations, potassium, and phosphorus are positively correlated with nodulation incidence. The two genera were generally similar in nodulation responses to soil and environmental factors.Support for portions of this study was provided by the National Science Foundation (PCM-8204885). Any opinions, findings conclusions or recommendations expressed are those of the authors and do not necessarily reflect the views of the sponsor. Oregon Agricultural Experiment Station technical paper number 7293.     

Variation with soil depth,topographic position and host species in the capacity of soils from an Australian locale to nodulateCasuarina andAllocasuarina seedlings

Sandy alluvial soils in a floodplain supporting a native stand ofCasuarina cunninghamiana Miq. produced about three times as many nodulated seedlings and more than twice as many nodules per nodulated seedling on roots of baitedCasuarina spp. than did clay loam red earth soils from the adjacent valley slope. Moist and well-aerated subsurficial alluvial sands had the greatest nodulation capacity of all the soils sampled. For all topographic positions, soil samples from depths greater than 20 cm promoted 76% more nodulated Casuarina seedlings than samples from the surficial 20 cm.Seedlings of three provenances ofC. cunninghamiana, together with seedlings ofC. glauca Sieb. ex Spreng.,C. cristata F. Muell ex Miq. andC. obesa Miq. developed significantly more nodules per pot and nodules per nodulated seedling in soils from this locale than seedlings of twoCasuarina equisetifolia Forst. provenances. Seedlings of two provenances ofAllocasuarina torulosa (Ait.) L. Johnson had fewer than 1% nodulated seedlings, a significantly lower level by far than that ofCasuarina seedlings.A. torulosa provenances also had significantly fewer nodulated seedlings per pot and nodules per nodulated seedling than all Casuarina hosts excepting one poorly-nodulated provenance ofC. equisetifolia.Nodulated seedlings of allCasuarina species had the capacity to fix atmospheric N₂, as indicated by acetylene-reduction capability. The presence of yellow cladodes and low rates of acetylene reduction per plant forC. cristata Miq. suggest that this association was poorly effective.     

Freeze/thaw stress in<Emphasis Type="Italic"> Ceanothus</Emphasis> of southern California chaparral

Freeze/thaw stress was examined in chaparral shrubs of the genus Ceanothus to determine the interactive effects of freezing and drought and to consider which is the more vulnerable component, the living leaves (symplast) or the non-living water transport system (apoplast). We hypothesized that where Ceanothus species co-occurred, the more inland species C. crassifolius would be more tolerant of low temperatures than the coastal species C. spinosus, both in terms of leaf survival (LT(50), or the temperature at which there is 50% loss of function or viability) and in terms of resistance to freezing-induced embolism (measurements of percent loss hydraulic conductivity due to embolism following freeze/thaw). Cooling experiments on 2 m long winter-acclimated shoots resulted in LT(50) values of about -10 degrees C for C. spinosus versus -18 degrees C for C. crassifolius. Freeze-thaw cycles resulted in no change in embolism when the plants were well hydrated (-0.7 to -2.0 MPa). However, when plants were dehydrated to -5.0 MPa, C. spinosus became 96% embolized with freeze/thaw, versus only 61% embolism for C. crassifolius. Stems of C. crassifolius became 90% and 97% embolized at -6.6 and -8.0 MPa, respectively, meaning that even in this species, stems could be more vulnerable than leaves under conditions of extreme water stress combined with freeze/thaw events. The dominance of C. crassifolius at colder sites and the restriction of C. spinosus to warmer sites are consistent with both the relative tolerance of their symplasts to low temperatures and the relative tolerance of their apoplasts to freeze events in combination with drought stress.     

Infectivity variation of Discaria trinervis-nodulating Frankia in Patagonian soil according to season and storage conditions

Changes in the infectious capacity of soilborne Frankia from the same site may depend on environmental conditions. To test this, we examined the effect of season of sampling, sample storage protocol and storage time. The nodulation capacity of Frankia from rhizospheric soils of Discaria trinervis (Hook et Arn.) Reiche (Rhamnaceae) growing in northwest Patagonia (Argentina) was measured using the most probable number method. Soil samples were collected seasonally and either stored moist at 4°C or air dried at room temperature for few days. Old (air-dried) soil samples were also assayed. All soils nodulated D. trinervis seedlings. Nodulation units (NU) ranged from 44 (spring, moist storage) to about 1 ml⁻¹ of soil (summer moist, and summer and autumn, air-dried storage), with intermediate values in other samples. Soils stored for 12 years, 6 months or 1 week had similar NU. Frankia NU positively correlated with soil water content ( r = 0.6, P < 0.05); therefore, it is likely that soil moisture is a relevant factor regulating soilborne Frankia nodulation ability in Patagonian soils. We suggest that Frankia can remain as spores or grow saprophytically in Patagonian soils.     

Nodulation and nitrogen fixation in Purshia: inoculation responses and species comparisons

Summary Nitrogen fixing trees and shrubs may be useful in revegetation efforts. Speculation that insufficient endophyte populations in surface soils may limit non-leguminous symbiotic nitrogen fixation in marginal land was explored.Purshia tridentata andP. glandulosa seedlings were grown in greenhouse trials using ten soils from nativePurshia sites. Treatments include a control, an inoculated treatment, and six mmole nitrogen amendment. When inoculated with aP.tridentata crushed nodule inoculum, two of five non-nodulating soils and three sparsely nodulating soils produced well nodulated plants. Inoculation also increased nodule mass, total nitrogen, nitrogen content and shoot dry mass in plants from some of the soils. Of the three soils failing to produce nodulated plants when inoculated, one produced plants that responded well to nitrogen additions but failed to nodulate under low nitrogen conditions; another produced severely stunted plants indicating nutritional limitations on the host; and the third produced plants that were not nitrogen deficient. An application of nitrogen completely suppressed nodulation in all but one soil. The twoPurshia species were similar in nodulation, nitrogen fixation and growth, although important exceptions exist that indicate species may differ in adaptability to certain soil conditions.     

Effects of continuous nitrogen application and nitrogen preconditioning on nodulation and growth ofCeanothus griseus varhorizontalis

Rooted cuttings ofCeanothus griseus varhorizontalis were irrigated with 0, 10, 20, 50, 75 or 100ppm nitrogen as NH₄NO₃ for eight weeks prior to inoculation with infectiveFrankia. After inoculation, half of the plants for each treatment nitrogen level continued to be irrigated with the preconditioning nitrogen level and half were given no more supplemental nitrogen. For plants continuously receiving nitrogen, nodule initiation (nodule number) was inversely correlated with increasing supplemental nitrogen levels, and suppressed above 50 ppm N. Leaf nitrogen above 2% in continuous-N plants correlated with greatly reduced or suppressed nodulation. Plants maintained after inoculation without supplemental nitrogen showed influence of the prior nitrogen treatment on nodulation. Preconditioning at 50 ppm and above greatly reduced the number of nodules formed. The evidence suggests that stored internal nitrogen can regulate nodulation.Plant biomass accumulated maximally when nodulation was suppressed, at 75 and 100 ppm supplemental N applied continuously. Internode elongation during the nodulation period occurred only on nodulated plants, or in the presence of supplemental N (10 ppm and above).     

Tissue water relations of three chaparral shrub species after wildfire

Summary We compared the tissue water relations among resprouts and seedlings of three chaparral species during the first summer drought after wildfire. Two of the species, Rhus laurina and Ceanothus spinosus recover after fire by a combination of resprouting and seedling establishment (facultative resprouters), whereas a third species, Ceanothus megacarpus recovers by seedling establishment alone (obligate seeder). Our objectives were to document any differences in tissue water characteristics that might arise between resprouts and seedlings and to test the hypothesis that seedlings of obligate seeders develop more drought tolerant characteristics of their tissues than seedlings of facultative resprouters. We found that resprouts had much higher predawn values of water potential, osmotic potential, and turgor potentials than seedlings. Predawn turgor potentials of resprouts were 1.5 MPa through July and August when turgor potentials for seedlings remained near 0 MPa. During summer months, midday water potentials were 2 to 3 MPa higher for resprouts than seedlings and midday conductances of resprouts were two to five fold greater than those of seedlings. Even though resprouts did not experience severe water stress like seedlings, their tissue water characteristics, as determined by pressure-volume curve analyses, were similar by the peak of the drought in August. Further-more, the tissue water characteristics of seedlings from the obligate seeder, C. megacarpus, were similar to those of facultative resprouters — R. laurina, and C. spinosus. We attribute the observed differences in plant water status between resprouts and seedlings to differences in rooting depths and access to soil moisture reserves during summer drought. We conclude that the higher growth rates, photosynthetic performance, and survivorship of postfire resprouts are primarily a result of higher water availability to resprouting tissues during summer months. It appears that the greater seedling survivorship during summer drought observed for the obligate seeder, C. megacarpus, is not associated with more favorable tissue water characteristics.     

Early colonization of red alder and Douglas fir by ectomycorrhizal fungi and Frankia in soils from the Oregon coast range

The potential for mycorrhizal formation and Frankia nodulation were studied in soils from six sites in the Pacific Northwest. The sites included young and old alder stands, a 1-year-old conifer clear-cut, a young conifer plantation, and rotation-aged and old-growth conifer stands. A bioassay procedure was used with both red alder and Douglas fir seedlings as hosts. After 6 weeks growth, seedlings of both hosts were harvested every 3 weeks for 21 weeks and numbers of nodules and ectomycorrhizal types estimated. Nodules formed on red alder and ectomycorrhizae formed on both alder and Douglas fir in soil from all sites. Nodulation potential was highest in soil from the alder stands and the conifer plantation. Seven morphologically distinct ectomycorrhizal types were recovered on Douglas fir and five on alder. Only Thelephora terrestris, a broad-host-range mycobiont, formed mycorrhizae on both hosts. New ectomycorrhizal types formed on both hosts throughout the bioassay. Ectomycorrhizal colonization of alder was greatest in the alder and clear-cut soils. Low ectomycorrhizal colonization on alder was found in soils from sites where conifers were actively growing. Ectomycorrhizal colonization of Douglas fir was highest in the young alder and conifer plantation soils and was low in the rotation-aged conifer soil. The highest diversity of ectomycorrhizal types was found on alder in the conifer clear-cut soil and on Douglas fir in the rotation-aged conifer soil. Effects of host specificity, nodulation and mycorrhiza-forming potential and nodule-mycorrhiza interactions on seedling establishment are discussed in relation to seral stage dynamics and attributes of pioneer ectomycorrhizal fungal species.     

Influence of soil temperature on niacin and thiamine in honey mesquite

Summary The influence of soil temperature was examined on niacin and thiamine concentration in honey mesquite (Prosopis glandulosa var.glandulosa) seedlings. The seedlings were grown in soil temperature regimes of 21, 27, and 32°C in a controlled environment growth room. Nodulation randomly occurred on the roots of the seedlings, necessitating separate analysis according to the occurrence of nodulation. Roots of nodulated seedlings from the 21°C soil temperature regime contained greater quantities of niacin and thiamine compared to root samples from seedlings grown in either 27 or 32°C regimes. Niacin concentration of non-nodulated seedlings was highest in samples from seedlings grown in the 27°C soil temperature regime and lowest in samples from seedlings grown in the 21°C regime. Thiamine concentration was the greatest from non-nodulated seedlings grown in the 27°C soil temperature regime, while the thiamine concentration of non-nodulated samples from the 32°C regime was the least. Optimal soil temperature for honey mesquite root growth appears to be about 27°C. At sub-optimal soil temperatures niacin might have limited ‘growth’ while at supra-optimal soil temperatures, thiamine might be a limiting factor. College of Agricultural Sciences Contribution No. T-9-164.     

Effects of Brassmosteroids on Growth, Nodulation, Phytohormone Content and Nitrogenase Activity in French Bean Under Water Stress

Effect of pre-treatments of 1 and 5 M epibrassinolide or homobrassinolide prior to water stress induction on changes in root nodulation and contents of endogenous abscisic acid (ABA) and cytokinin trans-zeatin riboside (ZR), and nitrogenase activity was investigated in the nodulated roots of Phaseolus vulgaris L. cv. Arka Suvidha. Brassinosteroids in the unstressed plants increased root nodulation, ZR content and nitrogenase activity, and also ameliorated their stress-induced decline in the nodulated roots. The ABA contents in the nodules of control or stressed plants were not altered by brassinosteroids treatment. There was an increase in pod yield by brassinosteroids treatment (5 M) in the irrigated control as well as stressed plants without influencing pod number or pod length. Among the brassinosteroids, epibrassinolide was relatively more effective.     

The capacity of Jamaican mine spoils, agricultural and forest soils to nodulate Myrica cerifera, Leucaena leucocephala and Casuarina cunninghamiana

Soils from seven sites on the island of Jamaica were assayed for the symbiotic diazotrophs Frankia and Rhizobium using serial dilutions. Most probable number and least squares regression methods were used to estimate each soil's capacity to nodulate native Myrica cerifera, exotic Leucaena leucocephala and exotic Casuarina cunninghamiana. The sample sites included a montane forest, a slash-and-burn agricultural site, reclaimed bauxite mining areas, abandoned sugar cane fields, and a garden plot. None of the host plants used in the bioassay were present on the sites sampled except for scattered L. leucocephala on one site. Frankia capable of nodulating M. cerifera, which is native to Jamaican highlands, occurred at all sites sampled. No C. cunninghamiana-infective Frankia was detected in soils sampled. Only soils from one site on the tropical coastal plain harbored rhizobia able to nodulate L. leucocephala (37 nodulation units cm^?3 of soil). A subset of nodulated M. cerifera and L. leucocephala reduced acetylene to ethylene indicating nitrogenase activity. The slash-and-burn agricultural site, which was situated at an elevation of 200 m and possessed both high natural fertility and high soil moisture-supplying capacity, had significantly greater Myrica infectious capacity (1 000 nodulation units cm^?3 of soil) than the other sites (7?207 nodulation units cm^?3 of soil). A planned, paired comparison revealed that a recently cultivated sugar cane field and a recently reclaimed bauxite mining site together had significantly less Myrica-infective Frankia (4 nodulation units cm^?3 of soil) than a corresponding pair of sites consisting of a sugar cane field abandoned for 25 years and a bauxite mining site reclaimed 20 years before sampling (118 nodulation units cm^?3 of soil). Results indicate that Myrica-infective Frankia is widespread in Jamaica, that the number of Myrica-infective Frankia units vary from site to site in accordance with soil type and soil history, that Jamaican sites sampled lack soil Frankia populations capable of nodulating a casuarina host, that rhizobial symbionts capable of nodulating L. leucocephala may be geographically restricted to lowlands in Jamaica, and that the occurrence of Frankia in these soils is independent of host plant presence.     

Nodulation ofMedicago sativa in solution culture

Summary The time course of increase in nodule number was observed in solution cultures of controlled composition, over periods of 5 to 7 weeks following the inoculation of 8-day-old seedlings. The plants produced successive crops of nodules two to three weeks apart. Maintaining nitrate at concentrations of 0.2 or 0.5mM greatly reduced the number of nodules in the first crop but only slightly or not at all in the larger following crops, relative to the numbers in nil-nitrate control cultures. This was not due to any decrease in the sensitivity of nodulation to inhibition by nitrate. Nodulation in the nil-nitrate controlplants was restrained following their abundant first-crop nodulation. The experiments ceased to be simple tests of effects of nitrate on rate of nodule production as soon as the first crop of nodules appeared. Subsequent nodule production was then inhibited approximately equally, by prior nodulation in the abundantly nodulated control-plants, and by nitrate in the sparsely nodulated plants in the nitrate treatment.When the effect of nitrate was tested on plants comparable in size and in number of previously established nodules, it inhibited nodule production by 70–80%, whether the plants were seedlings or month-old plants, previously nodulated or not.     

Frankia inoculation,soil biota,and host tissue amendment influence Casuarina nodulation capacity of a tropical soil

The effects of soil biota, Frankia inoculation and tissue amendment on nodulation capacity of a soil was investigated in a factorial study using bulked soil from beneath a Casuarina cunninghamiana tree and bioassays with C. cunninghamiana seedlings as capture plants. Nodulation capacities were determined from soils incubated in sterile jars at 21 °C for 1, 7, and 28 days, after receiving all combinations of the following treatments: ± steam pasteurization, ± inoculation with Frankia isolate CjI82001, and ± amendment with different concentrations of Casuarina cladode extracts. Soil respiration within sealed containers was determined periodically during the incubation period as a measure of overall microbial activity. Soil respiration, and thus overall microbial activity, was positively correlated with increasing concentrations of Casuarina cladode extracts. The nodulation capacity of soils inoculated with Frankia strain Cj82001 decreased over time, while those of unpasteurized soils without inoculation either increased or remained unaffected. The mean nodulation capacity of unpasteurized soil inoculated with Frankia CjI82001 was two to three times greater than the sum of values for unpasteurized and inoculated pasteurized soils. Our results suggest a positive synergism between soil biota as a whole and Frankia inoculum with respect to host infection.     

Frankia in acid soils of forests devoid of actinorhizal plants

The capacity of some acid forest soils to induce nodulation on a hybrid between Alnus incana (L.) Moench and A. glutinosa (L.) Gaertn. was investigated. Soil was sampled from tree stands devoid for decades of actinorhizal hosts. Seven-week-old Alnus seedlings growing m liquid culture were inoculated with soil dilutions. The nodules were counted after 6 weeks and classified as Sp⁻, if they lacked spores, or as Sp⁺. if spores were present, according to microscopy of microtome sections. Frankia was found in all the forest soils studied, apart from a soil from a Betula swamp. The highest nodulation capacities on Alnus , caused predominantly by Frankia of the Sp⁻ type. were observed in mineral soil sites with Betula stands — even higher than in soil from an A. incana stand. A positive correlation was found between the pH and the noduiation capacity of the soil.     

Rhizobial ecology of the woody legumePsorothamnus spinosus in a Sonoran Desert Arroyo

Psorothamnus spinosus seedlings were selected at random and excavated from 10×30 m quadrats along a 270 m transect in a southern Californian desert arroyo. Forty-six of the fifty seedlings excavated (92%) were nodulated. The mean number of nodules/plant was 7.0±3.6. Maximum rooting depth was 40 cm, and most (58.8%) of the nodules were found between 15 and 30 cm depth. The mean root/shoot ratio was 1.4±0.4. The rhizobial population density between 10 to 40 cm depth across all quadrats was non-detectable using a plant infection, most probable number method. All 70 isolates from field nodules were fast-growingRhizobium spp. Two colony morphologies were observed: wet, translucent, and wet, opaque. All isolates tested were effective on bothP. spinosus and another desert woody legume, mesquite (Prosopis glandulosa). Many isolates had a high relative efficiency, and seven of eight efficient isolates had uptake-hydrogenase activity. Root stimulation of the low rhizobial population found in the arroyo and subsequent root nodulation may be an important factor in seedling establishment by woody legumes in N-limited desert arroyo systems.     

Nodulation and nitrogen fixation (Acetylene reduction) in desert ironwood (Olneya tesota)

Summary Acacia greggi, Cercidium floridium, and Olneya tesota seeds were inoculated with soil from beneath mature native desert trees and grown in the greenhouse on a nitrogen free media. Olneya tesota seedlings nodulated and reduced acetylene to ethylene. Nodulation or acetylene reduction was not observed in A. greggi or C. floridium. This is the first report of nodulation and nitrogen fixation in Olneya tesota.     

Are Northwestern Patagonian “mallín” wetland meadows reservoirs of <Emphasis Type="BoldItalic">Ochetophila trinervis</Emphasis> infective <Emphasis Type="BoldItalic">Frankia?</Emphasis>

“Mallín” (plural mallines) is a particular kind of wetland occurring in Patagonian steppe and forests. In Northwest Patagonia, mallines are humid meadows with high net primary production. It was previously found that a mallín soil in the steppe devoid of actinorhizal plants had a higher Frankia nodulation capacity in Ochetophila trinervis (sin. Discaria trinervis) than other soils in the region. Under the hypothesis that mallín wetland meadows are reservoir of infective Frankia, we studied the Frankia nodulation capacity in O. trinervis of 12 mallín and their neighbouring steppe soils, by using plant bioassays. A qualitative plant bioassay showed that infective Frankia was present in most soils. The number of nodules per plant in seedlings inoculated with mallín soils was negatively correlated with soil water content while the opposite was true for plants inoculated with soils from neighbouring steppe. A quantitative bioassay was performed with eight representative soils, selected according to the number of nodules per plant produced in the qualitative assay and to the presence or not of different actinorhizal plants at the sites. Frankia nodulation units per cm³ of soil (NU) in mallín soils were higher than those in steppe. Water and organic matter content of soils were correlated with the higher nodulation capacity of mallines, which may account for the saprotrophic growth of Frankia in soils. The symbiosis was effective in plants inoculated with all soil samples. These results suggest that Northwestern Patagonian mallín wetland meadows are reservoirs of infective and effective Frankia propagules in O. trinervis.     

Distribution and diversity of rhizobia nodulating agroforestry legumes in soils from three continents in the tropics

The natural rhizobial populations of Calliandra calothyrsus, Gliricidia sepium, Leucaena leucocephala and Sesbania sesban were assessed in soils from nine sites across tropical areas of three continents. The rhizobial population size varied from undetectable numbers to 1.8 x 104 cells/g of soil depending on the trap host and the soil. Calliandra calothyrsus was the most promiscuous legume, nodulating in eight soils, while S. sesban nodulated in only one of the soils. Polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analyses of the 16S rRNA gene and the internally transcribed spacer (ITS) region between the 16S and 23S rRNA genes were used to assess the diversity and relative abundance of rhizobia trapped from seven of the soils by C. calothyrsus, G. sepium and L. leucocephala. Representatives of the 16S rRNA RFLP groups were also subjected to sequence analysis of the first 950 base pairs of the 16S rRNA gene. Eighty ITS groups were obtained, with none of the ITS types being sampled in more than one soil. RFLP analysis of the 16S rRNA yielded 23 'species' groups distributed among the Rhizobium, Mesorhizobium, Sinorhizobium and Agrobacterium branches of the rhizobial phylogenetic tree. The phylogeny of the isolates was independent of the site or host of isolation, with different rhizobial groups associated with each host across the soils from widely separated geographical regions. Although rhizobial populations in soils sampled from the centre of diversity of the host legumes were the most genetically diverse, soil acidity was highly correlated with the diversity of ITS types. Our results support the hypothesis that the success of these tree legumes in soils throughout the tropics is the result of their relative promiscuity (permissiveness) allowing nodulation with diverse indigenous rhizobial types.     

Effects of soil acidity on nodulation ofAlnus glutinosa and viability ofFrankia

Summary Black alder seedlings were grown from seed for 7 weeks in six soils limed to various pH levels and inoculated withFrankia in two inoculation-seeding time combinations (inoculated and seeded concurrently; inoculated then seeded 5 weeks after inoculation). Three mine soils and three non-mine soils were used. Soil pHs in the study ranged from 3.6 to 7.6. In the second inoculation-seeding time combination, a series of soil samples at each of the pH levels below 7.0 were relimed to pH 7.0 immediately prior to seeding. The purpose of the study was to examine the effects of soil acidity on the nodulation of black alder byFrankia and the viability ofFrankia in acid soils. Based on the average number of nodules established per seedling, soil pH was determined to be a significant factor affecting nodulation in the mine soils. The highest levels of nodulation occurred between soil pH 5.5 and 7.2. Below pH 5.5, nodulation was reduced. There was also evidence of decreased viability of the endophyte below pH 4.5.