Actinorhizal Species Research Articles

Identification of potential transcriptional regulators of actinorhizal symbioses in Casuarina glauca and Alnus glutinosa.

BackgroundTrees belonging to the Casuarinaceae and Betulaceae families play an important ecological role and are useful tools in forestry for degraded land rehabilitation and reforestation. These functions are linked to their capacity to establish symbiotic relationships with a nitrogen-fixing soil bacterium of the genus Frankia. However, the molecular mechanisms controlling the establishment of these symbioses are poorly understood. The aim of this work was to identify potential transcription factors involved in the establishment and functioning of actinorhizal symbioses.ResultsWe identified 202 putative transcription factors by in silico analysis in 40 families in Casuarina glauca (Casuarinaceae) and 195 in 35 families in Alnus glutinosa (Betulaceae) EST databases. Based on published transcriptome datasets and quantitative PCR analysis, we found that 39% and 26% of these transcription factors were regulated during C. glauca and A. glutinosa-Frankia interactions, respectively. Phylogenetic studies confirmed the presence of common key transcription factors such as NSP, NF-YA and ERN-related proteins involved in nodule formation in legumes, which confirm the existence of a common symbiosis signaling pathway in nitrogen-fixing root nodule symbioses. We also identified an actinorhizal-specific transcription factor belonging to the zinc finger C1-2i subfamily we named CgZF1 in C. glauca and AgZF1 in A. glutinosa.ConclusionsWe identified putative nodulation-associated transcription factors with particular emphasis on members of the GRAS, NF-YA, ERF and C2H2 families. Interestingly, comparison of the non-legume and legume TF with signaling elements from actinorhizal species revealed a new subgroup of nodule-specific C2H2 TF that could be specifically involved in actinorhizal symbioses. In silico identification, transcript analysis, and phylogeny reconstruction of transcription factor families paves the way for the study of specific molecular regulation of symbiosis in response to Frankia infection.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-014-0342-z) contains supplementary material, which is available to authorized users.

First report on the occurrence of the uncultivated cluster 2 Frankia microsymbionts in soil outside the native actinorhizal host range area

The occurrence of uncultivated Frankia was evaluated in Tunisian soils by a plant-trapping assay using Coriaria myrtifolia seedlings. Despite the lack of this compatible host plant for more than two centuries, soil-borne Frankia cells were detected in one sampled soil as shown by the development of root nodules on 2-year-old seedlings. Based on glnA sequences, Tunisian trapped Frankia strains belong to the uncultivated cluster 2 strains that associate with other Coriaria species and also with Ceanothus, Datisca and Rosaceae actinorhizal species. This is the first report on the occurrence of Frankia cluster 2 strains in soils from areas lacking compatible host plant groups.

The independent acquisition of plant root nitrogen-fixing symbiosis in Fabids recruited the same genetic pathway for nodule organogenesis.

Only species belonging to the Fabid clade, limited to four classes and ten families of Angiosperms, are able to form nitrogen-fixing root nodule symbioses (RNS) with soil bacteria. This concerns plants of the legume family (Fabaceae) and Parasponia (Cannabaceae) associated with the Gram-negative proteobacteria collectively called rhizobia and actinorhizal plants associated with the Gram-positive actinomycetes of the genus Frankia. Calcium and calmodulin-dependent protein kinase (CCaMK) is a key component of the common signaling pathway leading to both rhizobial and arbuscular mycorrhizal symbioses (AM) and plays a central role in cross-signaling between root nodule organogenesis and infection processes. Here, we show that CCaMK is also needed for successful actinorhiza formation and interaction with AM fungi in the actinorhizal tree Casuarina glauca and is also able to restore both nodulation and AM symbioses in a Medicago truncatula ccamk mutant. Besides, we expressed auto-active CgCCaMK lacking the auto-inhibitory/CaM domain in two actinorhizal species: C. glauca (Casuarinaceae), which develops an intracellular infection pathway, and Discaria trinervis (Rhamnaceae) which is characterized by an ancestral intercellular infection mechanism. In both species, we found induction of nodulation independent of Frankia similar to response to the activation of CCaMK in the rhizobia-legume symbiosis and conclude that the regulation of actinorhiza organogenesis is conserved regardless of the infection mode. It has been suggested that rhizobial and actinorhizal symbioses originated from a common ancestor with several independent evolutionary origins. Our findings are consistent with the recruitment of a similar genetic pathway governing rhizobial and Frankia nodule organogenesis.

Silencing of the chalcone synthase gene inCasuarina glaucahighlights the important role of flavonoids during nodulation

Nitrogen-fixing root nodulation is confined to four plant orders, including > 14,000 Leguminosae, one nonlegume genus Parasponia and c. 200 actinorhizal species that form symbioses with rhizobia and Frankia bacterial species, respectively. Flavonoids have been identified as plant signals and developmental regulators for nodulation in legumes and have long been hypothesized to play a critical role during actinorhizal nodulation. However, direct evidence of their involvement in actinorhizal symbiosis is lacking. Here, we used RNA interference to silence chalcone synthase, which is involved in the first committed step of the flavonoid biosynthetic pathway, in the actinorhizal tropical tree Casuarina glauca. Transformed flavonoid-deficient hairy roots were generated and used to study flavonoid accumulation and further nodulation. Knockdown of chalcone synthase expression reduced the level of specific flavonoids and resulted in severely impaired nodulation. Nodule formation was rescued by supplementing the plants with naringenin, which is an upstream intermediate in flavonoid biosynthesis. Our results provide, for the first time, direct evidence of an important role for flavonoids during the early stages of actinorhizal nodulation.

Use of Frankia and actinorhizal plants for degraded lands reclamation.

Degraded lands are defined by soils that have lost primary productivity due to abiotic or biotic stresses. Among the abiotic stresses, drought, salinity, and heavy metals are the main threats in tropical areas. These stresses affect plant growth and reduce their productivity. Nitrogen-fixing plants such as actinorhizal species that are able to grow in poor and disturbed soils are widely planted for the reclamation of such degraded lands. It has been reported that association of soil microbes especially the nitrogen-fixing bacteria Frankia with these actinorhizal plants can mitigate the adverse effects of abiotic and biotic stresses. Inoculation of actinorhizal plants with Frankia significantly improves plant growth, biomass, shoot and root N content, and survival rate after transplanting in fields. However, the success of establishment of actinorhizal plantation in degraded sites depends upon the choice of effective strains of Frankia. Studies related to the beneficial role of Frankia on the establishment of actinorhizal plants in degraded soils are scarce. In this review, we describe some examples of the use of Frankia inoculation to improve actinorhizal plant performances in harsh conditions for reclamation of degraded lands.

Genome Sequence of “Candidatus Frankia datiscae” Dg1, the Uncultured Microsymbiont from Nitrogen-Fixing Root Nodules of the Dicot Datisca glomerata

Members of the noncultured clade of Frankia enter into root nodule symbioses with actinorhizal species from the orders Cucurbitales and Rosales. We report the genome sequence of a member of this clade originally from Pakistan but obtained from root nodules of the American plant Datisca glomerata without isolation in culture.

Early signaling in actinorhizal symbioses.

Nitrogen-fixing root nodulation, confined to four plant orders, encompasses more than 14,000 Leguminosae species, and approximately 200 actinorhizal species forming symbioses with rhizobia (Rhizobium, Bradyrhizobium, etc) and Frankia bacterial species, respectively. While several genetic components of the host-symbiont interaction have been identified in legumes, little is known about the genetic bases of actinorhizal symbiosis. However, we recently demonstrated the existence of common symbiotic signaling pathways in actinorhizals and legumes. Moreover, important data on the identification of flavonoids as plant signaling compounds and the role for auxins during Frankia infection process and nodule organogenesis have been acquired. All together these results lead us to propose a unified model for symbiotic exchange and genetic control of actinorhizal symbiosis.

Symbiotic Signaling in Actinorhizal Symbioses

Actinorhizal symbioses are mutualistic associations between plants belonging to eight angiosperm families and soil bacteria of the genus Frankia. These interactions lead to the formation of new root organs, actinorhizal nodules, where the bacteria are hosted and fix atmospheric nitrogen thus providing the plant with an almost unlimited source of nitrogen for its nutrition. It involves an elaborate signaling between both partners of the symbiosis. In recent years, our knowledge of this signaling pathway has increased tremendously thanks to a series of technical breakthroughs including the sequencing of three Frankia genomes [1] and the implementation of RNA silencing technology for two actinorhizal species. In this review, we describe all these recent advances, current researches on symbiotic signaling in actinorhizal symbioses and give some potential future research directions.

Reproductive ecology of Ochetophila trinervis in Northwest Patagonia.

Native actinorhizal species Ochetophila trinervis (Gillies ex Hook. & Arn.) Poepp. ex Miers, also known as Discaria trinervis (Kellermann et al. 2005), grows along watercourses and distant from them, along a rainfall gradient in north-west Patagonia. We studied the reproductive ecology of this species under different macro- and micro-environmental conditions, in three zones in a rainfall gradient (western, intermediate and eastern) with two sites for each one, near and distant to a watercourse (riparian and dry-land). We performed field studies and germination trials. Plant size, reproductive effort (seed abundance per branch) and seed bank size of O. trinervis, were favoured by the proximity of streams in the drier environments of the gradient. The abundance of seedlings and saplings in the field was very low, which was in agreement with a lack of germination in the field, despite the good germination capacity of seeds. Sexual reproduction of O. trinervis was affected by the low abundance of seedlings. Lower rainfall, higher temperatures and a longer growing season along the gradient favour the potential regeneration of the species. Because of its nitrogen-fixing capacity and other features, O. trinervis has potential for reclamation of eroded lands in Patagonia. Patterns of seed biology and regeneration presented in this study will aid in the use of O. trinervis in the reclamation of disturbed lands.

An overview of actinorhizal plants in Africa.

A compilation and synthesis of information derived from plant databases and other sources on the occurrence, diversity and geographic distribution of actinorhizal plants in Africa is presented in this review. Actinorhizal plants are a specific group of non-leguminous, woody dicots having symbiotic, nitrogen-fixing root nodules that are induced on roots of actinorhizal plant species by soil actinomycetes of the genus Frankia. There is a lack of basic information on actinorhizal plants in Africa compared with other major land masses in the world. Results indicate that most, if not all, African countries and climatic regions have native or introduced actinorhizal species. A synthesis of available information indicates that there are six families, nine genera and 38 reported species of actinorhizal plants in Africa. Of these, 21 species are native and 17 are exotic. The families and corresponding number of species in each genus are: Betulaceae (native Alnus glutinosa (1), exotic Alnus (2)); Casuarinaceae (exotic Casuarina (5), exotic Allocasuarina (3), exotic Gymnostoma deplancheana (1)); Coriariaceae (native Coriaria myrtifolia (1)); Myricaceae (native Morella (19), exotic Morella cerifera (1)); Rhamnaceae (exotic Ceanothus caeruleus (1), exotic Colletia paradoxa (1)); and Elaeagnaceae (exotic Eleaegnus angustifolia (1)). Four reports of native, actinorhizal Ceanothus species in Africa found in the database were determined to be false, instead, being non-actinorhizal species. Widespread plantings of exotic Casuarinaceae have been introduced into tropical and arid zones of Africa as multipurpose trees, especially in arid regions where native species do not occur. There is a diverse assemblage of native species of Morella in Africa, mostly shrubs or small trees, which provide medicine, other useful chemicals and wildlife habitat. Many native Morella species are isolated in montane islands, apparently leading to greater speciation than in Eurasia from where the genus migrated into Africa. The current status and knowledge of African actinorhizal plants indicates a need to focus research on their biogeography, biology, ecology, genetics and use.

Jasmonate biosynthesis in legume and actinorhizal nodules

Jasmonic acid (JA) is a plant signalling compound that has been implicated in the regulation of mutualistic symbioses. In order to understand the spatial distribution of JA biosynthetic capacity in nodules of two actinorhizal species, Casaurina glauca and Datisca glomerata, and one legume, Medicago truncatula, we determined the localization of allene oxide cyclase (AOC) which catalyses a committed step in JA biosynthesis. In all nodule types analysed, AOC was detected exclusively in uninfected cells. The levels of JA were compared in the roots and nodules of the three plant species. The nodules and noninoculated roots of the two actinorhizal species, and the root systems of M. truncatula, noninoculated or nodulated with wild-type Sinorhizobium meliloti or with mutants unable to fix nitrogen, did not show significant differences in JA levels. However, JA levels in all plant organs examined increased significantly on mechanical disturbance. To study whether JA played a regulatory role in the nodules of M. truncatula, composite plants containing roots expressing an MtAOC1-sense or MtAOC1-RNAi construct were inoculated with S. meliloti. Neither an increase nor reduction in AOC levels resulted in altered nodule formation. These data suggest that jasmonates are not involved in the development and function of root nodules.

Characterization of microsatellite markers for black alder (Alnus glutinosa [L.] Gaertn)

Black alder (Alnus glutinosa Gaertn.) is an actinorhizal tree species that is becoming increasingly interesting as a pioneer species and a soil improvement species, as well as for its ecological benefits for river ecosystems and bank stabilisation. Little is known at the molecular level about its genetic variation and population structure. Microsatellite markers suitable for use for A. glutinosa were searched for by data mining the publicly available Alnus sequences and by testing cross-genera amplification of 26 available Corylus SSR markers. We report the characteristics of 11 microsatellite markers that are usable for A. glutinosa. The observed and expected heterozygosity ranged from 0.160 to 0.920 and 0.147 to 0.882, respectively. Two loci showed significant deviation from Hardy–Weinberg Equilibrium. This deviation is likely to be related to the presence of null alleles. These markers will be useful in genetic population studies and the preservation of A. glutinosa diversity.

Variation of 15N natural abundance in leaves and nodules of actinorhizal shrubs in Northwest Patagonia

This study was performed to assess the N2-fixing capability of the native actinorhizal species Ochetophila trinervis (sin. Discaria trinervis) and Discaria chacaye (Rhamnaceae) in Northwest Patagonia. We measured the N concentration and 15N natural abundance in leaves and nodules of O. trinervis and D. chacaye, in leaves of associated non-actinorhizal vegetation, and in the soils under each sampled plant. O. trinervis and D. chacaye had foliar N concentrations that were about twice that of non-actinorhizal shrubs growing at the same sites, even though soils varied four-fold in total N across the sites. Leaves of both actinorhizal plants had a similar δ15N at any site and were close to atmospheric values. The foliar δ15N of non-actinorhizal plants and soil δ15N were strongly correlated across the sites. Nodules were depleted in δ15N relative to the foliage of the respective actinorhizal species. In conjunction with the uniformly high foliage N concentration of these actinorhizal plants and the universal presence of vesicles observed in root nodules, these data strongly suggest that O. trinervis and D. chacaye obtain a significant amount of their N from N2 fixation. To calculate the proportion of N derived from atmosphere, theoretical B-values were estimated. In all cases where the δ15N of fixing and reference foliage were significantly different, O. trinervis and D. chacaye obtained almost all of their N from N2 fixation. These results are the first to demonstrate N2 fixation by O. trinervis and D. chacaye in the field and therefore suggest an important role for these actinorhizal plants in the N economy of ecosystems in northwest Patagonia as well as their potential use for restoration of degraded lands in this region.

Foliar‐Nitrogen and Phosphorus Resorption Patterns Differ among Nitrogen‐Fixing and Nonfixing Temperate‐Deciduous Trees and Shrubs

Nutrient resorption from senescing leaves enables plants to conserve nutrients. Low foliar‐nitrogen (N) resorption levels have been estimated in past studies for some N‐fixing woody plant taxa. To determine whether this trait is characteristic of N‐fixing temperate species, N‐resorption proficiency estimates were derived for several species during two seasons in central Illinois, United States. Nutrient‐resorption proficiencies were measured for 24 species of temperate‐deciduous woody plants. Thirteen actinorhizal species and three non‐N‐fixing species in families with actinorhizal species were sampled. One N‐fixing woody legume, four non‐N‐fixing legumes, and two species lacking N‐fixing capabilities and unrelated to these groups were also sampled. Nitrogen resorption, based on resorption proficiency values, was less in N‐fixing species (1.78% N retained in abscised leaves) than in non‐N‐fixing species (1.01% N). Phosphorus (P) resorption, however, was greater in N‐fixing species (0.06% P retained in abscised leaves) than in non‐N‐fixing species (0.26% P). Resorption of P may have been low in a group supporting ectomycorrhizal symbionts. Results indicate a consistent pattern in which N‐fixing species are less proficient at N resorption than other temperate‐deciduous woody species.

Nodulation and Growth of Alnus nitida and Alnus maritima Inoculated with Species-specific and Nonspecific Frankia

Abstract Actinorhizal plants form N2-fixing symbioses with soil-borne bacteria of the genus Frankia. Potential exists for development of sustainable, actinorhizal nursery crops that obtain most of their required N through N2 fixation, but information on host-symbiont specificity, presence of compatible Frankia in soils, and techniques to inoculate during plant production is lacking. Our objectives were to determine the effect of inoculum type and source and the effect of supplemental N on nodulation, growth, and N content of two actinorhizal species, Alnus nitida (Spach) Endl. and Alnus maritima (Marsh.) Muhl. ex Nutt. Plants of both species were subjected to one of four inoculum treatments (two crushed-nodule inocula: species-specific and cross inoculation, and two soil inocula: soil collected beneath native Alnus rubra Bong. in Washington state and native prairie soil from Iowa), were supplied fertilizer with or without N, and were grown in a greenhouse for 22 weeks. Inoculated plants nodulated, grew larger and faster, and accrued greater N content than uninoculated controls in both fertilizer treatments. Plants that received species-specific inoculum grew larger, acquired more dry weight from symbioses, and accumulated higher N content than cross-inoculated plants. Plants of A. nitida inoculated with soil from Washington state grew larger and accumulated more dry weight from symbioses than those inoculated with prairie soil, but A. maritima grew similarly with soil inoculum from both sources. Our results demonstrate that A. nitida and A. maritima can benefit from N2-fixing symbiosis during production and that potential exists for development of superior inocula and inoculation techniques.

Genetic Diversity of Frankia Microsymbionts from the Relict Species Myrica faya (Ait.) and Myrica rivas-martinezii (S.) in Canary Islands and Hawaii

In the Western Canary Islands, Myrica faya and Myrica rivas-martinezii (Myricaceae) are phylogenetically close, endemic, actinorhizal species presumed to be remnants either of the European or the African Tertiary floras. Unisolated Frankia strains from field-collected nodules on Tenerife, Gomera, and La Palma Islands were compared by their rrs gene and 16S-23S intergenic spacer (IGS) restriction patterns. To compare the genetic diversity of Frankia strains from within and outside the host's native range, nodules of M. faya field plants were collected both in Canary Islands and in Hawaii, where this species is an exotic invasive. Myrica rivas-martinezii, endemic to the Canary Islands, was sparsely nodulated in the field. Frankia strains harbored in field-collected nodules of M. faya and M. rivas-martinezii belonged to the Elaeagnaceae strains' genetic cluster and exhibited a high degree of diversity. Frankia genotypes were specific to each host species. In the Canary archipelago, we found no relationship between site of collection and Frankia genotype for M. faya. The only exceptions were strains from site 2 in Tenerife, a location with a geological history different from the other sites sampled. Hawaiian and Canarian M. faya strains had no genotypes in common, raising questions concerning the origin of M. faya-infective Frankia in Hawaii. Nodular strains of M. rivas-martinezii from nursery plants were genetically characterized and shown to be divergent from the strains of field-collected nodules and belong to the Alnus-Casuarina strains cluster. This suggests Myrica may have the potential to nodulate with a broader range of Frankia genotypes under artificial conditions than has been detected in field-collected nodules.

Location and Anatomy of Nodules on Alnus maritima Subjected to Flooding

Although many species of Alnus Miller grow in wet soils, none is as closely associated with low-oxygen, waterlogged soils as Alnus maritima (Marsh.) Muhl. ex Nutt. (seaside alder). An actinorhizal species with promise for use in horticultural landscapes, land reclamation, and sustainable systems, A. maritima associates with Frankia Brunchorst, thereby forming root nodules in which gaseous nitrogen is fixed. Our objective was to determine how root-zone moisture conditions influence the occurrence, location, and anatomy of nodules on A. maritima. Plants of Alnus maritima subsp. maritima Schrader and Graves were established in root zones with compatible Frankia and subjected to four moisture regimens (daily watered/drained, partially flooded, totally flooded, and totally flooded with argon bubbled through the flood water) for 8 weeks. Oxygen content of the root zone, number and location of nodules on root systems, and dry weight and nitrogen content of shoots were determined. Root-zone oxygen content ranged from 17.3 kPa for daily watered/drained plants to 0.9 kPa for argon-treated plants. Across all treatments, 87% of the nodules were within the upper one-third (4 cm) of the root zone. Although shoot dry weights of daily watered/drained and partially flooded plants were not different, daily watered/drained plants had more nitrogen in their leaves (2.53 vs. 2.21 mg·g-1). Nodulation occurred in all treatments, but nodules on totally flooded roots (with or without argon) were limited to a single lobe; in contrast, multilobed nodules were prevalent on partially flooded and daily watered/drained plants. Frankia infection within submerged nodule lobes was limited to one or two layers of cortical cells. Submerged nodules developed large air spaces between cortical cells, and phenolic-containing cells appeared to inhibit Frankia expansion within the nodule. These data suggest that access to root-zone oxygen is critical to the Frankia-A. maritima subsp. maritima symbiosis, and that plants of this subspecies in the drained soils of managed landscapes may benefit more than plants in native wetland habitats from nodulation and nitrogen fixation.

Adaptations of Alnus maritima Nodules to Low Oxygen in the Root Zone

Alnus maritima (Marsh.) Muhl. ex Nutt. is unique among alders in its degree of preference for low-oxygen soils of wetlands. An actinorhizal species with promise for use in sustainable horticulture, A. maritima develops a root-nodule symbiosis with nitrogen-fixing Frankia. Nodules of other actinorhizal species that are obligate wetland natives are adapted to low oxygen, and expression of hemoglobin is common to these taxa. Our objectives were to determine the range of oxygen tension under which Alnus maritima subsp. maritima fixes nitrogen and to investigate a potential role for hemoglobin in adaptation of nodules to low oxygen. Roots of plants, cultured aeroponically, were subjected to eight oxygen tensions from 0 to 32 kPa. After four weeks, plant dry weight, nodule fresh weight, nitrogenase activity, and photosynthetic rate were measured. In addition, nodules were assayed spectrophotometrically for the presence of hemoglobin. A quadratic function best described the influence of oxygen on plant dry weight, nodule fresh weight, nitrogenase activity, and photosynthetic rate with maximal values above 20 kPa. Alnus serrulata (Ait.) Willd. is sympatric with A. maritima subsp. maritima but is not an obligate inhabitant of wetlands. In a separate experiment, we found higher nitrogenase activity in A. maritima subsp. maritima than in A. serrulata (0.74 vs. 0.26 μmol/h per plant) at hypoxic oxygen tensions. Further, optical absorption spectra of nodule extracts confirmed hemoglobin within nodules of A. maritima subsp. maritima. Our data suggest that hemoglobin contributes to oxygen regulation in nodules of A. maritima subsp. maritima.

Novel expression pattern of cytosolic Gln synthetase in nitrogen-fixing root nodules of the actinorhizal host, Datisca glomerata.

Gln synthetase (GS) is the key enzyme of primary ammonia assimilation in nitrogen-fixing root nodules of legumes and actinorhizal (Frankia-nodulated) plants. In root nodules of Datisca glomerata (Datiscaceae), transcripts hybridizing to a conserved coding region of the abundant nodule isoform, DgGS1-1, are abundant in uninfected nodule cortical tissue, but expression was not detectable in the infected zone or in the nodule meristem. Similarly, the GS holoprotein is immunolocalized exclusively to the uninfected nodule tissue. Phylogenetic analysis of the full-length cDNA of DgGS1-1 indicates affinities with cytosolic GS genes from legumes, the actinorhizal species Alnus glutinosa, and nonnodulating species, Vitis vinifera and Hevea brasilensis. The D. glomerata nodule GS expression pattern is a new variant among reported root nodule symbioses and may reflect an unusual nitrogen transfer pathway from the Frankia nodule microsymbiont to the plant infected tissue, coupled to a distinctive nitrogen cycle in the uninfected cortical tissue. Arg, Gln, and Glu are the major amino acids present in D. glomerata nodules, but Arg was not detected at high levels in leaves or roots. Arg as a major nodule nitrogen storage form is not found in other root nodule types except in the phylogenetically related Coriaria. Catabolism of Arg through the urea cycle could generate free ammonium in the uninfected tissue where GS is expressed.

Nitrogen Fixation as a Stress-Avoidance Strategy Among Actinorhizal (Non-Legume) Trees and Shrubs

Summary Actinorhizal species of trees and shrubs are a diverse group of plants that may survive in poor soils by virtue of their associations with the actinomycete Frankia. These species include several important woody plants that are well suited for horticultural use in temperate climates. The symbiosis between Frankia and actinorhizal species shows some similarity to symbioses between rhizobia and woody legumes, and a common ancestor has been proposed for the predisposition to root-nodule symbiosis. Despite their probable common origin, important differences exist between actinorhizal and leguminous symbioses; characteristics of the microsymbiont, nodule architecture, and mechanisms controlling oxygen relations of the nodule are among the ways the two systems differ. If nitrogen fixation is sustained under unfavorable conditions, woody plants that associate with nitrogen-fixing organisms may show enhanced tolerance of environmental stress; species of plants capable of nitrogen-fixing symbioses are known to have comparatively strong resistance to invasion by pathogens. Expansion of the capacity to form nitrogen-fixing symbioses to novel species is a goal of those concerned with the economic and ecological impact of chemical fertilizers. Small in-roads have been made in this regard, but much remains to be discovered about introducing nitrogen fixation to additional species. Herein we review biological aspects of actinorhizal symbioses; consider the horticultural potential of temperate, woody species that form these symbioses; and discuss how nitrogen-fixing symbiosis may impact the stress resistance and use of actinorhizal species as horticultural crops.