Genetic diversity among Elaeagnus compatible Frankia strains and sympatric-related nitrogen-fixing actinobacteria revealed by nifH sequence analysis

Elaeagnus compatible Frankia isolates from Tunisian soil have been previously clustered with Frankia, colonizing Elaeagnaceae and Rhamnaceae in two different phylogenetic subgroups, while strain BMG5.6 was described as a new lineage closely related to Frankia and Micromonospora genera. In this study we further assess the diversity of captured Frankia and the relationship with BMG5.6-like actinobacteria, by using nifH gene sequences. Using PCR-RFLP screening on DNA extracted from lobe nodules, additional microsymbionts sharing BMG5.6 features have been detected proving a widespread occurrence of these actinobacteria in Elaeagnus root nodules. Neighbour-Joining trees of Frankia nifH sequences were consistent with previously published 16S rRNA and GlnII phylogenetic trees. Although four main clades could be discerned, actinobacterial strain BMG5.6 was clustered with Frankia strains isolated from Elaeagnus. The present study underscored the emanation of new diazotrophic taxon isolated from actinorhizal nodules occupying intermediate taxonomic position between Frankia and Micromonospora. Moreover, its aberrant position in nifH phylogeny should open network investigations on the natural history of nitrogen-fixing gene among actinobacteria.     

Reproductive biology and conservation implications of three endangered snapdragon species (Antirrhinum, Plantaginaceae)

About 32% of Antirrhinum species are considered to be endangered; however, no field studies have focused on their reproductive biology. In this work, several aspects of the reproductive biology (flowering phenology, floral biology, breeding system) and potential limits on seed quantity and quality (pollen limitation, inbreeding depression) were studied in natural populations of three endangered species of the genus (Antirrhinum charidemi, Antirrhinum subbaeticum, Antirrhinum valentinum). Results disclose that all three species need insect visitors for seed production since fruit set after autonomous self-pollination was lower than under hand cross-pollination. A. charidemi and A. valentinum were mainly self-incompatible, whereas A. subbaeticum was self-compatible but herkogamous. Supplementary pollination in open-pollinated flowers only increased fruit set and seed set relative to controls in a given population of A. valentinum. Preliminary data on inbreeding depression at early life-cycle stages of the self-compatible A. subbaeticum revealed that the cumulative level was low. Despite the three species being closely related and sharing many ecological characteristics, they show different mating systems, and different factors limit seed quantity and quality. Thus, caution should be taken when making a common conservation plan for a group of closely related taxa.     

Conserving marginal populations of the food plant (Impatiens noli-tangere) of an endangered moth (Eustroma reticulatum) in a changing climate

Impatiens noli-tangere is scarce in the UK and probably only native to the Lake District and Wales. It is the sole food plant for the endangered moth Eustroma reticulatum. Significant annual fluctuations in the size of I. noli-tangere populations endanger the continued presence of E. reticulatum in the UK. In this study, variation in population size was monitored across native populations of I. noli-tangere in the English Lake District and Wales. In 1998, there was a crash in the population size of all metapopulations in the Lake District but not of those found in Wales. A molecular survey of the genetic affinities of samples in 1999 from both regions and a reference population from Switzerland was performed using AFLP and ISSR analyses. The consensus UPGMA dendrogram and a PCO scatter plot revealed clear differentiation between the populations of I. noli-tangere in Wales and those in the Lake District. Most of the genetic variation in the UK (H_T=0.064) was partitioned between (G_ST=0.455) rather than within (H_S=0.034) regions, inferring little gene flow occurs between regions. There was similar bias towards differentiation between metapopulations in Wales, again consistent with low levels of interpopulation gene flow. This contrasts with far lower levels of differentiation in the Lake District which suggests modest rates of gene flow may occur between populations. It is concluded that in the event of local extinction of sites or populations, reintroductions should be restricted to samples collected from the same region. We then surveyed climatic variables to identify those most likely to cause local extinctions. Climatic correlates of population size were sought from two Lake District metapopulations situated close to a meteorological station. A combination of three climatic variables common to both sites explained 81-84% of the variation in plant number between 1990 and 2001. Projected trends for these climatic variables were used in a Monte Carlo simulation which suggested an increased risk of I. noli-tangere population crashes by 2050 at Coniston Water, but not at Derwentwater. Implications of these findings for practical conservation strategies are explored.     

Habitat use and preliminary demographic evaluation of the critically endangered Mediterranean monk seal (Monachus monachus) in the Cilician Basin (Eastern Mediterranean)

Over the last few decades a dramatic decline in the number of Mediterranean monk seals (Monachus monachus) limited scientific studies on the biology, behaviour, and ecology of the species. This lack of information impaired the effectiveness of conservation strategies. Thus, any further information gathered on the monk seal has utmost value in the work to halt and reverse the plight of the species. A hitherto unknown Cilician Basin colony of the Mediterranean monk seal has been investigated during a 6-year period between 1995 and 2001. Results of direct in-cave surveys and land based seal-watch observations enabled the evaluation of habitat use and preliminary demographic information. A total of 39 caves were discovered among which only three were used for breeding. Scarcity of the breeding caves emphasised the importance of breeding habitat in the survival of the colony. Common features of the breeding caves are found as an entrance with a protective barrier against strong waves; a chamber with a wide beach; and a well-sheltered shallow pool. Supplementary data obtained from infrared monitors installed in three actively used caves were used to assess the number of individuals. Throughout the study period, 25 individuals were identified and 11 newborn pups were found. Evaluation of the results suggests the presence of sub-regions inhabited by an individually identified sub-group of monk seals. Recommendations for conservation guided by this information is a Marine Protected Area consisting of two zones; a restricted core zone prohibiting any human activity securing breeding habitat and a restricted fishery zone securing sustainability of the food source.     

Relationship between population size and genetic diversity in endangered populations of the European bullhead (Cottus gobio): implications for conservation

This study investigated the relationship between the current size of endangered bullhead (Cottus gobio) populations and microsatellite genetic variability. Additionally, the microsatellite data were used to evaluate whether a genetic test for population bottlenecks was able to provide evidence of recent severe population declines. Finally, our results were used to develop conservation priorities and measures. Population size appears to be a crucial parameter in determining the amount of genetic diversity that can be preserved in bullheads, since a significant positive correlation was observed between both variables. Furthermore, in some populations we were able to detect genetic signatures of the documented decline in population size. We suggest that the most immediate goal for bullhead conservation should be to increase the size and the range of the populations, and in doing so minimise or even reverse further genetic erosion. Potential management actions like habitat quality improvement, reduction of river fragmentation and supplementation programmes (translocation, supportive breeding) are discussed.     

Assessing limitations on population growth in two critically endangered Acacia taxa

Flowering phenology, soil seedbank and the impact of fire, weeds and grazing were investigated in two rare Acacia taxa restricted to the agricultural district north of Perth Western Australia. Acacia aprica is known from six extant populations and one extinct population. Five of the extant populations are restricted to linear road reserves with the sixth population located in a small native vegetation remnant. Acacia cochlocarpa ssp. cochlocarpa is known from a single population also situated on a road verge. Size class structure, levels of canopy death and an absence of juveniles indicated that all populations are in decline. Flowering intensity and success varied between populations and years in A. aprica and between years in A. cochlocarpa ssp. cochlocarpa. Seed bank analysis indicated that seeds were patchily distributed while experimental fires demonstrated that such events could break seed dormancy and promote germination. Both taxa have similar germination physiologies and showed increased germination after seeds were exposed to heat. Following emergence, however, competition with annual weeds had a negative impact on seedling growth and survival in both taxa. While vertebrate grazing had some influence, weeds were the major inhibitory influence on recruitment. Reduced fire frequencies since fragmentation may be responsible for population decline in both taxa but other site specific factors such as weeds and grazing may affect the establishment of seedlings following fire.     

Keystone effects of the endangered Stephens' kangaroo rat (Dipodomys stephensi)

Kangaroo rats (Dipodomys) have been argued to exert keystone effects because they interact strongly with other species, and their removal results in major changes in community structure. We evaluated the nature and magnitude of such impacts by the endangered Stephens' kangaroo rat (Dipodomys stephensi), employing an exclosure experiment to determine how the removal of this species impacted relative cover by vegetation, plant species diversity, abundance of a dominant annual plant genus (Erodium), and predation of seeds from artificial trays. Fifteen months of D. stephensi removal resulted in significant increases in herb cover and Erodium abundance, and significant declines in bare ground and in seed predation. These results support the hypothesis that D. stephensi has major impacts on plant community structure and seems to function as a keystone species. It remains to be seen how the loss of this species from much of its range will influence local community composition and dynamics.     

Distribution and efficiency of Rhizobium leguminosarum strains nodulating Phaseolus vulgaris in Northern Spanish soils: Selection of native strains that replace conventional N fertilization

Phaseolus vulgaris is a legume extensively cultivated in Spain, León province being the most important producer. This province produces selected varieties of common bean highly appreciated by their quality that warrants a Protected Geographic Indication (PGI). In this work we analysed the rhizobia present in nodules of the variety “Riñón” in several soils from León province in order to select native rhizobial strains to be used as biofertilizers. The analysis of rrs and housekeeping genes of these strains showed that they belong to two phylogenetic groups within Rhizobium leguminosarum (I and II). Although the group II strains were most abundant in nodules, very effective strains were also found in group I. Strains LCS0306 from group I and LBM1123 from group II were the best nitrogen fixers among all strains isolated and were selected for field experiments. The field research showed that the biofertilization of common bean with native and selected rhizobial strains can completely replace the fertilization with chemical N fertilizers. The biofertiliser designed in such way, was valid for the whole agroecological area, regardless the specific properties of each soil and microclimatic conditions. This conclusion can be generalised as a strategy for the development of biofertilisers in different agroecological conditions worldwide.     

Ex situ genetic conservation of endangered Vatica guangxiensis (Dipterocarpaceae) in China

RAPD polymorphisms were applied to check the efficiency of ex situ genetic conservation of endangered Vatica guangxiensis X. L. Mo. endemic to southwestern China. Low level of genetic variation was revealed in three remaining natural populations. Twenty random primers, each with 10 base pairs, generated 231 bands with 53.68% being polymorphic, and with an average of 32.46% being polymorphic in each natural population. Strong population differentiation was revealed by AMOVA (analysis of molecular variance) and Gst value was 0.3764. The population ML ex situ conserved in the Xishuangbanna Tropical Botanical Garden contained an intermediate genetic variation compared with natural populations, with 30.74% bands being polymorphic. Of the total 231 bands generated in V. guangxiensis, 204 bands were also detected in population ML, indicating that 88.31% of the total genetic variations of this species were conserved in ex situ population. If only the alleles with moderate to high frequency (P>0.05) were considered, 204 out of 209 bands (97.61%) occurred in ex situ population ML. RAPD analysis also detected one exclusive band in natural population NS, and five in natural population NP, three of these exclusive bands were generated in every samples of natural population (NP), and other three had moderate to high frequencies. While none of these exclusive bands were detected in ex situ conserved population ML. Our conclusions are that the ex situ conserved population ML contains representative genetic variation to maintain long-term survival and evolutionary process of V. guangxiensis, and that more extensive ex situ sampling in natural population NS and NP is needed to conserve more exclusive alleles in ex situ population. The tropical area in the Botanical Garden would play a more important role in the ex situ conservation of rare and endangered plants.     

Reproductive constraints for the long-term persistence of fragmented Acacia dealbata (Mimosaceae) populations in southeast Australia

Fragmented and degraded vegetation characterises agricultural landscapes across southern Australian. Remnant vegetation within these regions performs a number of vital ecological and hydrological roles, but little is known about whether or how fragmentation is affecting the long-term persistence of these critical landscape elements. Acacias are a significant component of many remnant vegetation communities across Australia, forming numerous integral faunal and floral relationships. Here, reproductive output of 11 fragmented Acacia dealbata (Mimosaceae) populations from across the southern tablelands of New South Wales was assessed over 2 years to identify reproductive constraints associated with increasing vegetation fragmentation. Fertilization success is the major reproductive constraint, particularly in small populations, and probably reflects a self-incompatible reproductive strategy. During 2002 larger and more dense populations produced more legumes (p = 0.014 and <0.001, respectively) while in 2003 these two variables were associated with increased fertilization success (p = 0.004 and 0.017, respectively). There was also some suggestion that populations with fewer exotic species also experienced increased fertilization success (p = 0.055). Assessment of plant performance within populations suggests that consistent reproductive output of particular individuals within small populations may limit reproductive compatibility within these populations over time. The long-term persistence of many small A. dealbata populations may be jeopardised by low seed set, and limited recruitment and aging stands. Immediate steps are now required to ensure that these populations continue contributing to landscape function by augmenting populations, improving connectivity, and allowing disturbance events that will stimulate recruitment.     

Detecting hybridization in wild (Coturnix c. coturnix) and domesticated (Coturnix c. japonica) quail populations

Two subspecies of Coturnix coturnix have allopatric distributions in Europe, Maghreb and western Asia (the Common quail C. c. coturnix), and in eastern Asia (the Japanese quail C. c. japonica), except for sympatric breeding areas in the Baikal (Russia) and Kentei (Mongolia) regions, where they could hybridize. Japanese quails have been reproduced in captivity for centuries and domesticated. Massive releasing of Japanese domesticated quails, or hybrids, in west European countries might threaten with hybridization the gene pool of wild quail populations. Here, we used mitochondrial (mtDNA) and nuclear DNA markers (microsatellites), and Bayesian admixture analyses aiming to assess subspecies distinction and identify hybrids in wild and captive-reared quails. A phylogenetic tree identified two major mtDNA clades, corresponding to the two subspecies, and allowed detecting 12 hybrids in wild Common quails sampled in Spain. Additional hybrids were identified in wild quails sampled in Spain, Italy and Senegal using microsatellite markers. Wild quails sampled in Mongolia showed japonica mtDNAs, but their microsatellite genotypes were admixed, suggesting that Common and Japanese quails can hybridize in nature. Some captive-reared stocks of Japanese quails were also admixed. Introgression of domesticated Japanese quail genes in wild Common quail populations might affect the phenotypic expression of functional traits, as body size, feather colours, sexual calls and migratory behaviours. Hence, restocking with captive-reared non-native quails should be banned.     

Two genetically distinct units of the Chinese sika deer (cervus nippon): analyses of mitochondrial DNA variation

Chinese sika deer, Cervus nippon, are currently threatened by habitat loss, fragmentation and human hunting, which has led to the extinction of three subspecies in the wild. The remaining subspecies subsist in the narrow regions of Jilin, Heilongjiang, Sichuan, Gansu, Jiangxi, Anhui, and Zhejiang provinces. In order to design effective conservation strategies for the Chinese sika deer, we have investigated genetic diversity, population structure and gene flow in the Chinese sika deer populations by analyzing ≈995 base pairs of the mitochondrial DNA (mtDNA) control region in 59 individuals sampled from the northeast of China, Sichuan, Jiangxi and Zhejiang. Chinese sika deer exhibited low mtDNA diversity and high gene flow among the four populations, and showed no strong geographical structure. The analyses of mtDNA variation among individuals of sika deer identified only two main phylogenetic groups even though three subspecies were sampled. These data singles out the Zhejiang population as being highly genetically distinct and worthy of separate conservation consideration. Therefore, it is recommended that a breeding program for the Zhejiang population be established.     

Pre-symbiotic and symbiotic interactions between Glomus intraradices and two Paenibacillus species isolated from AM propagules. In vitro and in vivo assays with soybean (AG043RG) as plant host

Two indole-producing Paenibacillus species, known to be associated with propagules of arbuscular mycorrhizal (AM) fungi, were examined for their mycorrhization helper bacteria activity at pre-symbiotic and symbiotic stages of the AM association. The effects were tested under in vitro and in vivo conditions using an axenically propagated strain of the AM fungus Glomus intraradices and Glycine max (soybean) as the plant host. The rates of spore germination and re-growth of intraradical mycelium were not affected by inoculation with Paenibacillus strains in spite of the variation of indole production measured in the bacterial supernatants. However, a significant promotion in pre-symbiotic mycelium development occurred after inoculation of both bacteria under in vitro conditions. The Paenibacillus rhizosphaerae strain TGX5E significantly increased the extraradical mycelium network, the rates of sporulation, and root colonization in the in vitro symbiotic association. These results were also observed in the rhizosphere of soybean plants grown under greenhouse conditions, when P. rhizosphaerae was co-inoculated with G. intraradices. However, soybean dry biomass production was not associated with the increased development and infectivity values of G. intraradices. Paenibacillus favisporus strain TG1R2 caused suppression of the parameters evaluated for G. intraradices during in vitro symbiotic stages, but not under in vivo conditions. The extraradical mycelium network produced and the colonization of soybean roots by G. intraradices were promoted compared to the control treatments. In addition, dual inoculation had a promoting effect on soybean biomass production. In summary, species of Paenibacillus associated with AM fungus structures in the soil, may have a promoting effect on short term pre-symbiotic mycelium development, and little impact on AM propagule germination. These findings could explain the associations found between some bacterial strains and AM fungus propagules.     

Livestock predation by endangered African wild dogs (Lycaon pictus) in northern Kenya

Most large mammalian carnivores are in global decline, principally due to conflict with livestock farmers. Because endangered African wild dogs (Lycaon pictus) range widely, often beyond the boundaries of protected areas, they may be particularly exposed to lethal control by farmers, even where nominally protected by reserves. Hence, effectively conserving wild dogs demands resolution of their conflicts with farmers. We investigated livestock depredation by African wild dogs living outside protected areas in northern Kenya. Scat analysis confirmed the distribution of depredation events reported by local farmers, indicating that farmer reports - collated by local Community Liaison Officers - gave a reasonably good index of the true pattern of depredation. Although livestock were abundant throughout the study area, depredation was exceedingly uncommon (approximately one attack per 1000 km² per year) and the costs of tolerating wild dogs were very low (US $3.40/wilddog/year) where wild prey remained, even at low densities. However, where wild prey were seriously depleted, wild dogs killed livestock repeatedly, and the cost of sustaining them rose to US $389/wilddog/year. Hence, although wild dogs had a negligible economic impact in the region, their impact was locally severe. Conservation activities for wild dogs are most likely to be successful if targeted at areas where wild prey remain, and where traditional herding practices have been retained.     

Screening of bacterial isolates from various European soils for in vitro antagonistic activity towards Rhizoctonia solani and Fusarium oxysporum: Site-dependent composition and diversity revealed

A cultivation-based approach was used to determine the in vitro antagonistic potential of soil bacteria towards Rhizoctonia solani AG3 and Fusarium oxysporum f. sp. lini (Foln3). Four composite soil samples were collected from four agricultural sites with previous documentation of disease suppression, located in France (FR), the Netherlands (NL), Sweden (SE) and the United Kingdom (UK). Similarly, two sites from Germany (Berlin, G-BR; and Braunschweig, G-BS) without documentation of disease suppression were sampled. Total bacterial counts were determined by plating serial dilutions from the composite soil samples onto R2A, AGS and King's B media. A total of 1,788 isolates (approximately 100 isolates per medium and site) was screened for antifungal activity, and in vitro antagonists (327 isolates) were found amongst the dominant culturable bacteria isolated from all six soils. The overall proportion of antagonists and the number of isolates with inhibitory activity against F. oxysporum were highest in three of the suppressive soils (FR, NL and SE). Characterization of antagonistic bacteria revealed a high phenotypic and genotypic diversity. Siderophore and protease activity were the most prominent phenotypic traits amongst the antagonists. The composition and diversity of antagonists in each soil was site-specific. Nevertheless, none of the antimicrobial traits of bacteria potentially contributing to soil suppressiveness analyzed in this study could be regarded as specific to a given site.     

Functional conservation units for the endangered Alcon Blue butterfly Maculinea alcon in Belgium (Lepidoptera: Lycaenidae)

To organize and prioritise species-specific conservation efforts, we delineate `functional conservation units' for the threatened Alcon Blue butterfly Maculinea alcon in Belgium. We used detailed distribution data on the butterfly, its host plant and its habitat, present-day population sizes and its mobility and colonization capacity to determine functional conservation units (FCUs) on different spatial scales: FCU-1, i.e., the 12 presently occupied habitat patches plus the area within a range of 500 m surrounding them (the maximum local movement distance, based on mark-release-recapture data), FCU-2, i.e., the areas within a range of 2 km around the occupied habitat patches (the maximum observed colonization capacity) and FCU-3, i.e., potential re-introduction sites (sites where M. alcon went extinct recently). We suggest different management and planning measures for each type of functional conservation unit and discuss translocation and re-introduction as `intensive care' conservation measures for this threatened and sedentary species.     

Distribution, status, and conservation of radiated tortoises (Geochelone radiata) in Madagascar

The radiated tortoise, Geochelone radiata, one of Madagascar’s four endemic tortoises, occupies a narrow band of xeric spiny forest along the island’s southwest coast. Traditionally avoided by indigenous tribes, these tortoises are now routinely harvested for food. An accurate assessment of human exploitation remains problematic, however, hindered by limited, dated statistics available on tortoise populations. To update the radiated tortoise’s status and distribution, we established a series of line transects at seven localities across its range and implemented a mark-recapture study at one of these localities (Cap Sainte Marie). Tortoises currently range from south of Tulear to east of Cap Sainte Marie, at density estimates spanning 27-5744 tortoises/km². The mark-recapture estimate for Cap Sainte Marie (1905-2105 tortoises/km²) was substantially higher than its transect estimate (654 tortoises/km²) though comparable to actual tortoise captures (1438) there. Thus, our transect density values probably err as underestimates, and from these data, we calculate a conservative total population size of 12 million radiated tortoises. We also examined mitochondrial DNA sequences (ND4 gene) for two individuals/locality in a preliminary assessment of genetic variation across the species’ range. Only two ND4 haplotypes were recovered, the more common haplotype representing 13 of the 14 individuals. We offer several conservation recommendations in light of our survey results.     

Cryptic diversity in the well-studied terrestrial worm Cognettia sphagnetorum (Clitellata: Enchytraeidae)

The terrestrial worm Cognettia sphagnetorum has been used as a model in several studies focusing on research areas such as climate change as well as forest and soil ecology; it has also been shown to play a key role in the decomposition of organic matter and nutrient cycling. Cognettia is an enchytraeid genus commonly found in acidic terrestrial habitats, such as coniferous forests and bogs. In this study, the diversity of the genus, with particular focus on the morphospecies C. sphagnetorum in northern Europe, is assessed using four molecular markers, the mitochondrial COI (cytochrome c oxidase subunit I) and16S (16S ribosomal RNA), and the nuclear H3 (Histone 3) and ITS (Internal Transcribed Spacer). The datasets were first delimited into Molecular Operational Units (MOTUs) and the existence of global barcoding-gaps was tested. Single gene-trees were then estimated for all genes using Bayesian Inference, and a species tree was estimated with all markers combined using the multi-species coalescence. The results show that in northern Europe the genus consists of at least eight MOTUs supported by all genes except H3. Four of these MOTUs were within the morphotaxon C. sphagnetorum and two within Cognettia glandulosa. C. sphagnetorum s.l. was found to be non-monophyletic in all gene-trees, as well as in the species tree. As the MOTUs were well separated and non-monophyly was observed within C. sphagnetorum s.l., we conclude that the MOTUs are best treated as separate species. Given that cryptic diversity was found in this genus, we recommend that material of Cognettia used in future studies should be identified using molecular barcodes.     

Habitat of endangered white abalone, Haliotis sorenseni

Surveys with a submersible at offshore islands and banks in southern California found that white abalone were most abundant at depths between 43 and 60 m. This is deeper than estimates taken when white abalone were more abundant. Densities were highest at sites far from fishing ports. Controlling for depth and site found that white abalone were significantly more abundant in areas with Laminaria farlowii (an alga) but abalone were not associated with areas high in the cover of other algae (Pelagophycus porra or Eisenia arborea) or the amount of sand in the habitat (except that abalone always occurred on rock). Within an area with abalone, the particular rock they occurred on was significantly larger than unoccupied neighboring rocks. Occupied rocks were not significantly different in algal cover or in sea urchin density than unoccupied neighboring rocks. The position of abalone on a rock was nearer to the rock-sand interface than would be expected based on a random distribution. More white abalone were feeding when in association with red urchins, perhaps because both grazers capture drift algae to eat. These data may aid future efforts to locate white abalone brood stock and identify locations for outplanting.     

Growth promoting effects of corn (Zea mays) bacterial isolates under greenhouse and field conditions

Fertilizer costs are a major component of corn production. The use of biofertilizers may be one way of reducing production costs. In this study we present isolation and identification of three plant growth promoting bacteria that were identified as Enterobacter cloacae (CR1), Pseudomonas putida (CR7) and Stenotrophomonas maltophilia (CR3). All bacterial strains produced IAA in the presence of 100 mg l⁻¹ of tryptophan and antifungal metabolites to several soilborne pathogens. S. maltophilia and E. cloacae had broad spectrum activity against most Fusarium species. The only strain that was positive for nitrogen fixation was E. cloacae and it, and P. putida, were also positive for phosphate solubilization. These bacteria and the corn isolate Sphingobacterium canadense CR11, and known plant growth promoting bacterium Burkholderia phytofirmans E24 were used to inoculate corn seed to examine growth promotion of two lines of corn, varieties 39D82 and 39M27 under greenhouse conditions. When grown in sterilized sand varieties 39M27 and 39D82 showed significant increases in total dry weights of root and shoot of 10-20% and 13-28% and 17-32% and 21-31% respectively. Plants of the two varieties grown in soil collected from a corn field had respective increases in dry weights of root and shoot of 10-30% and 12-35% and 11-19% and 10-18%. In sand, a bacterial mixture was highly effective whereas in soil individual bacteria namely P. putida CR7 and E. cloacae CR1 gave the best results with 39M27 and 39D82 respectively. These isolates and another corn isolate, Azospirillum zeae N7, were tested in a sandy soil with a 55 and 110 kg ha⁻¹ of nitrogen fertility at the Delhi research Station of Agriculture and Agri-Food Canada over two years. Although out of seven bacterial treatments, no treatment provided a statistically significant yield increase over control plots but S. canadense CR11 and A. zeae N7 provided statistically significant yield increase as compared to other bacteria. The 110 kg rate of nitrogen provided significant yield increase compared to the 55 kg rate in both years.