Potential biocontrol actinobacteria: Rhizospheric isolates from the Argentine Pampas lowlands legumes.

Control of fungal plant diseases by using naturally occurring non-pathogenic microorganisms represents a promising approach to biocontrol agents. This study reports the isolation, characterization, and fungal antagonistic activity of actinobacteria from forage soils in the Flooding Pampa, Argentina. A total of 32 saprophytic strains of actinobacteria were obtained by different isolation methods from rhizospheric soil of Lotus tenuis growing in the Salado River Basin. Based on physiological traits, eight isolates were selected for their biocontrol-related activities such as production of lytic extracellular enzymes, siderophores, hydrogen cyanide (HCN), and antagonistic activity against Cercospora sojina, Macrophomia phaseolina, Phomopsis sp., Fusarium oxysporum, and Fusarium verticilloides. These actinobacteria strains were characterized morphologically, physiologically, and identified by using molecular techniques. The characterization of biocontrol-related activities in vitro showed positive results for exoprotease, phospholipase, fungal growth inhibition, and siderophore production. However, none of the strains was positive for the production of hydrogen cyanide (HCN). Streptomyces sp. MM140 presented the highest index for biocontrol, and appear to be promising pathogenic fungi biocontrol agents. These results show the potential capacity of actinobacteria isolated from forage soils in the Argentine Pampas lowlands as promising biocontrol agents, and their future agronomic applications.

Datathons: fostering equitability in data reuse in ecology.

Approaches to rapidly collecting global biodiversity data are increasingly important, but biodiversity blind spots persist. We organized a three-day Datathon event to improve the openness of local biodiversity data and facilitate data reuse by local researchers. The first Datathon, organized among microbial ecologists in Uruguay and Argentina assembled the largest microbiome dataset in the region to date and formed collaborative consortia for microbiome data synthesis.

Analysis of nodulation kinetics in Frankia-Discaria trinervis symbiosis reveals different factors involved in the nodulation process.

The induction of root nodule development in actinorhizal symbiosis would depend on the concentration of factors produced by the bacteria and the plant. A detailed analysis of nodulation description parameters revealed different factors related to the nodulation process. The initial time for nodulation (t(0)), the initial nodulation rate (v(0)) and the total time of nodule development (t(NOD)) were defined and consequently quantified in different experimental conditions: co-inoculation of Discaria trinervis with increasing concentrations of different non-infective bacteria together with the full compatible infective Frankia strain (the indicator strain) used at a limiting concentration or by changing plant factor(s) concentration. All the above nodulation parameters were modified by changing doses of full compatibility infective strain Frankia BCU110501; v(0) appears to be an expression of symbiotic recognition between partners as only fully symbiotic indicator Frankia BCU110501 was able to change it; t(0) seems not to reflect symbiotic recognition because it can also be modified by non-infective Frankia but suggest the existence of a basic level of plant microbe recognition. The initial time for nodulation t(0), reflecting the time required for the early interactions toward nodulation, is an inverse measure of the ability to establish early interactions toward nodulation. The increase in plant factors concentration also reduces t(0) values, suggesting that a plant factor is involved and favors very early interactions. Increases in plant factors concentration also modify the final number of nodules per plant and the nodule cluster profile along the taproot as an expression of the autoregulation phenomenon. Meanwhile, Frankia inoculums' concentration, either infective or not, modified t(NOD) in an opposite way plant factors did. In conclusion, the analysis of nodulation kinetics appears to be an appropriate tool to investigate factors involved in the symbiotic interaction leading to the formation of nitrogen-fixing nodules.

Draft Genome Sequence of Burkholderia cordobensis Type Strain LMG 27620, Isolated from Agricultural Soils in Argentina.

Bacteria of the genus Burkholderia are commonly found in diverse ecological niches in nature. We report here the draft genome sequence of Burkholderia cordobensis type strain LMG 27620, isolated from agricultural soil in Córdoba, Argentina. This strain harbors several genes involved in chitin utilization and phenol degradation, which make it an interesting candidate for biocontrol purposes and xenobiotic degradation in polluted environments.

Time course of nodule development in the Discaria trinervis (Rhamnaceae) -Frankia symbiosis.

The time course of initiation and development of root nodules was investigated in the South American actinorhizal shrub Discaria trinervis (Rhamnaceae). A local strain of Frankia (BCU110501) which was isolated from D. trinervis nodules, was used as inoculum. Inoculated seedlings were periodically studied under the light microscope after clearing with aqueous NaClO. In parallel, semithin and ultrathin sections were analysed by light and electron microscopy. Infection by Frankia BCU110501 involved intercellular penetration among epidermal and cortical root cells. Nodule primordia were detected from 6 d after inoculation, while bacteria were progressing through intercellular spaces of the outer layers of cortical cells. Invasion of host cells by the symbiont occurred 7-9 d after inoculation, and hypertrophy of the primordium cells was associated with Frankia penetration. Root hairs were not deformed during the early events of nodule formation. From 13 to 16 d after inoculation, the proximal cellular zone of the primordia behaved differently from the other tissues after NaClO treatment and remained darkly pigmented. At the same time, differentiation of Frankia vesicles started to occur inside already infected cells. By 16 d after inoculation, spherical vesicles of BCU110501 were homogeneously distributed in the host cells. These vesicles were septate and surrounded by void space. Frankia spores or sporangia were not observed in the nodule tissue. This study has clarified the mode of Frankia penetration in D. trinervis, one of the Rhamnaceae which also includes Ceanothus. The events involved in infection, nodule induction, host-cell infection and vesicle differentiation have been characterized and identified as time-segregated developmental processes in the ontogeny of D. trinervis root nodules.

Diffusible factors from Frankia modify nodulation kinetics in Discaria trinervis, an intercellular root-infected actinorhizal symbiosis.

Frankia BCU110501 induces nitrogen-fixing root nodules in Discaria trinervis (Gillies ex Hook. & Arn.) Reiche (Rhamnaceae) via intercellular colonisation, without root hair deformation. It produces diffusible factors (DFs) that might be involved in early interactions with the D. trinervis roots, playing a role in the nodulation process. The induction of root nodule development in actinorhizal symbiosis would depend on the concentration of factors produced by the bacteria and the plant. A detailed analysis of nodulation kinetics revealed that these DFs produce changes at the level of initial rate of nodulation and also in nodulation profile. Diluted Frankia BCU110501 inoculum could be activated in less than 96h by DFs produced by Frankia BCU110501 cells that had been previously washed. Biochemical characterisation showed that Frankia BCU110501 DFs have a molecular weight of <12kDa, are negatively charged at pH 7.0 and seem to contain a peptide bond necessary for their activity. Frankia BCU110501, belonging to Frankia Clade 3, does not induce nodules in Alnus acuminata H.B.K. ssp. acuminata but is able to deform root hairs, as do Frankia strains from Clade 1. The root hair deforming activity of Frankia BCU110501 DFs show the same biochemical characteristics of the DFs involved in nodulation of D. trinervis. These results suggest that Frankia symbiotic factors have a basic structure regardless of the infection pathway of the host plant.

Diffusible factors involved in early interactions of actinorhizal symbiosis are modulated by the host plant but are not enough to break the host range barrier.

Nodulation kinetics were analysed in two nitrogen-fixing actinorhizal symbioses that show different pathways for infection: Alnus acuminata H. B. K., which is infected by Frankia ArI3, and Discaria trinervis (Hooker et Arnot) Reiche, which is infected by Frankia BCU110501. Both pairs are incompatible in cross-inoculation experiments. The dose-response effects in nodulation were studied in A. acuminata seedlings using different concentrations of compatible and incompatible bacteria in co-inoculation experiments. Restriction fragment length polymorphism PCR analysis and plant-trapping analysis showed no co-occupation in A. acuminata nodules when plants were co-inoculated with Frankia BCU110501 and Frankia ArI3. Despite the lack of co-occupation, the noninfective BCU110501 could modify the nodulation parameters of the non-host A. acuminata when infective ArI3 was present in the inoculum. The results suggest that although BCU110501 was not able to induce nodulation in A. acuminata, its interaction with the plant could induce autoregulation as if some level of infection or partial recognition could be achieved. We explored the possibility that physiological complementation of the heterologous Frankia BCU110501 for nodulation of A. acuminata originated in the homologous Frankia ArI3 in the presence of compatible root exudates. Despite the possibility of full activation between bacteria and the host, there was no co-infection of Frankia BCU110501 in Alnus or of Frankia ArI3 in Discaria either. These negative results suggest a physical recognition barrier in actinorhizal symbiosis that operates after early interactions, involving something other than root exudates and diffusible factors of bacterial or plant origin, regardless of the infection pathway.

Infectivity of soilborne Frankia and mycorrhizae in Discaria trinervis along a vegetation gradient in Patagonian soil.

The infective capacities of the nitrogen fixing Actinomycete Frankia and arbuscular mycorrhizal fungi from soils near watercourses, along a vegetation gradient, were studied using plant bioassays. Frankia and arbuscular mycorrhizas capable of infecting Discaria trinervis were found at seventeen sites sampled. More specific enumeration of the infective capacities of both microorganisms in relation to environmental factors was performed in seven representative soils of the analysed vegetation zones (rainforest, xeric forest and steppe) using the most probable number method. The highest nodulation capacities ranged from 340 infective units g(-1 )soil, in a steppe marsh devoid of actinorhizas, to 61 in a coastal actinorhizal scrub (in xeric forest). The highest number of infective mycorrhizal units--also found in marsh--was 145. In general, rainforest soils had the lowest values for both microorganisms. Infective units of Frankia and arbuscular mycorrhizal fungi in soil were positively correlated (r = 0.89, P < 0.05). Both soilborne symbionts showed the highest infective capacity in semi-arid conditions nearby watercourse and at the valley bottom location. Tripartite symbiosis was effective in plants inoculated with steppe and xeric forest soils and plants inoculated with Frankia BCU110501 and Glomus mosseae. Interaction between both symbionts and influence of environmental conditions, in general, would contribute to define comparable trends of their infective capacities.

Regulation of nodulation in the absence of N2 is different in actinorhizal plants with different infection pathways.

Root nodulation in actinorhizal plants, like Discaria trinervis and Alnus incana, is subject to feedback regulatory mechanisms that control infection by Frankia and nodule development. Nodule pattern in the root system is controlled by an autoregulatory process that is induced soon after inoculation with Frankia. The final number of nodules, as well as nodule biomass in relation to plant biomass, are both modulated by a second mechanism which seems to be related to the N status of the plant. Mature nodules are, in part, involved in the latter process, since nodule excision from the root system releases the inhibition of infection and nodule development. To study the effect of N(2) fixation in this process, nodulated D. trinervis and A. incana plants were incubated under a N(2)-free atmosphere. Discaria trinervis is an intercellularly infected species while A. incana is infected intracellularly, via root hairs. Both symbioses responded with an increment in nodule biomass, but with different strategies. Discaria trinervis increased the biomass of existing nodules without significant development of new nodules, while in A. incana nodule biomass increased due to the development of nodules from new infections, but also from the release of arrested infections. It appears that in D. trinervis nodules there is an additional source for inhibition of new infections and nodule development that is independent of N(2) fixation and nitrogen assimilation. It is proposed here that the intercellular Frankia filaments commonly present in the D. trinervis nodule apex, is the origin for the autoregulatory signals that sustain the blockage of initiated nodule primordia and prevent new roots from infections. When turning to A. incana plants, it seems likely that this signal is related to the early autoregulation of nodulation in A. incana seedlings and is no longer present in mature nodules. Thus, actinorhizal symbioses belonging to relatively distant phylogenetic groups and displaying different infection pathways, show different feedback regulatory processes that control root nodulation by Frankia.