Analysis of rhizobial endosymbionts of Vicia, Lathyrus and Trifolium species used to maintain mountain firewalls in Sierra Nevada National Park (South Spain)

Villadas, Pablo J.; Lasa, Ana V.; Martínez‐Hidalgo, Pilar; Flores-Félix, José David; Martínez‐Molina, Eustoquio; Toro, Nicolás; Velázquez, Encarna; Fernández‐López, Manuel

doi:10.1016/j.syapm.2016.11.008

Cited by 11 publications

(17 citation statements)

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“…Extracted RNA was treated with DNase I (Roche, Germany) and RNase Out Recombinant Ribonuclease Inhibitor (Invitrogen, CA, USA) and the absence of DNA was confirmed by the lack of amplification of 16S rRNA gene, according to Villadas et al . 54 cDNA was synthesized using random primers and SuperScript TM II Reverse Transcriptase (Invitrogen, CA, USA) as recommended by the manufacturer. Double stranded cDNA was synthesized using RNase H (Roche), DNA polymerase I (Promega) and E. coli DNA ligase (Invitrogen, CA, USA).…”

Section: Methodsmentioning

confidence: 99%

Metabarcoding reveals that rhizospheric microbiota of Quercus pyrenaica is composed by a relatively small number of bacterial taxa highly abundant

Lasa

Fernández-González

Villadas

et al. 2019

Sci Rep

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Melojo oak (Quercus pyrenaica Willd.) is a key tree species of Mediterranean forests; however, these forests show an advanced stage of deterioration in the Iberian Peninsula. Plant-associated microorganisms play an essential role improving their host’s fitness, hence, a better understanding of oak rhizospheric microbiome, especially of those active members, could be the first step towards microbiome-based approaches for oak-forest improvement. Here we reported, for the first time, the diversity of total (DNA-based) and potentially active (RNA-based) bacterial communities of different melojo-oak forest formations through pyrosequencing of 16S rRNA gene amplicons. We found that potentially active bacterial communities were as rich and diverse as total bacterial communities, but different in terms of relative abundance patterns in some of the studied areas. Both core microbiomes were dominated by a relatively small percentage of OTUs, most of which showed positive correlation between both libraries. However, the uncoupling between abundance (rDNA) and potential activity (rRNA) for some taxa suggests that the most abundant taxa are not always the most active, and that low-abundance OTUs may have a strong influence on oak’s rhizospheric ecology. Thus, measurement of rRNA:rDNA ratio could be helpful in identifying major players for the development of bacterial bioinoculants.

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Section: Methodsmentioning

confidence: 99%

Metabarcoding reveals that rhizospheric microbiota of Quercus pyrenaica is composed by a relatively small number of bacterial taxa highly abundant

Lasa

Fernández-González

Villadas

et al. 2019

Sci Rep

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“…Regarding seed yield, only V. disperma in Soportújar responded to mycorrhizal inoculation. On the other hand, there was no significant effect associated with rhizobium inoculation for any of the species studied, probably because this type of bacteria are already present in the soil, and hence inoculation had no additional benefit (Villadas et al 2016).…”

Section: Discussionmentioning

confidence: 90%

“…In Lanjarón, the experimental layout was a randomized block design with four blocks and with four treatments per block and species: rhizobium, sheep-penning (SP), and sheep-penning + mycorrhizae (SPM), and control. The rhizobium treatment consisted of pelletization of the seeds with native rhizobial species and symbiovars that were identified and isolated by Villadas et al (2016) in the same area for these legume species. Sheep penning was carried out as follows: in summer 2014, 300 sheep were fenced into a 300 m 2 pen for 16 hours a day over 3 days.…”

Section: Methodsmentioning

confidence: 99%

“…Firstly, symbiotic nodulating bacteria, collectively termed rhizobia, in legume roots ensure nitrogen availability for the plant due to their capability to reduce atmospheric nitrogen to ammonia in symbiotic root nodules (Leigh 2002). Furthermore, when using local ecotypes, local endosymbionts are usually present in the soil, but if necessary, seeds should be inoculated before sowing to ensure their persistence (Villadas et al 2016). Secondly, vesicular-arbuscular mycorrhizae (VAM) are known to be very efficient in improving growth and nitrogen content in legumes, due to their ability to improve phosphorus intake by plants, and increase the supply of other immobilized nutrients or those present in low concentrations, such as ammonium, zinc and copper (Barea et al 1987;Barea et al 2005).…”

Section: Introductionmentioning

confidence: 99%

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Potential of wild annual legumes for mountain pasture restoration at two silvopastoral sites in southern Spain: promising species and soil-improvement techniques

et al. 2019

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This study evaluates the potential for pasture improvement and restoration at two silvopastoral sites. We used a total of 16 wild legume species under different management systems (rhizobial inoculation, mycorrhizal inoculation, sheep penning, and sheep penning with mycorrhizal inoculation), at two mountain sites in Sierra Nevada Natural Park (Granada, southern Spain). The first site, Soportújar, had higher soil fertility and rainfall than the second, Lanjarón. Forage yields in Soportújar ranged from 265 to 8970 kg DM ha-1 , Vicia amphicarpa, Vicia monantha, Vicia disperma and Lathyrus clymenum being the most productive species. Mycorrhizal inoculation resulted in higher forage yields for Lens nigricans, V. disperma and Vicia lutea. Seed yields were low, ranging from 5.9 to 1234 kg ha-1. Forage yields in Lanjarón were lower, ranging from 46 to 1415 kg DM ha-1 ; and the most productive species were V. monantha, V. disperma, Lathyrus cicera and Medicago rigidula. Sheep penning alone and together with mycorrhizal treatment resulted in greater forage yields for most of the species studied, although differences were only significant for V. disperma, V. monantha and L. sphaericus. Seed yields ranged from 0.4 to 60 kg ha-1. In conclusion, we recommend V. Post-print: Agroforestry Systems (2019) 1-13 monantha, V. disperma and L. cicera followed by V. amphicarpa and L. clymenum (in wetter more fertile sites) and M. rigidula (in drier sites), as they seem to be the best adapted to the pedoclimatic conditions of Sierra Nevada Natural Park. Increasing nutrients in the soil (by sheep penning) and promoting nutrient assimilation (by mycorrhizal inoculation) may be effective strategies for increasing pasture biomass in silvopastoral sites.

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“…; Villadas et al . ). This also proved to be the case with the L. linifolius rhizobial isolates, which all segregated with nodA sequences for R. leguminosarum biovar viciae , including the sequenced type strain 3841 (Fig.…”

Section: Discussionmentioning

confidence: 97%

Towards a characterisation of the wild legume bitter vetch (Lathyrus linifolius L. (Reichard) Bässler): heteromorphic seed germination, root nodule structure and N‐fixing rhizobial symbionts

et al. 2018

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Lathyrus linifolius L. (Reichard) Bässler (Fabiaceae, bitter vetch) is a nitrogen (N) fixing species. A coloniser of low nutrient (N) soils, it supports biodiversity such as key moth and butterfly species, and its roots are known for their organoleptic and claimed therapeutic properties. Thus, the species has high potential for restoration, conservation, novel cropping and as a model species. The last because of its genetic synteny with important pulse crops. However, regeneration and functional attributes of L. linifolius remain to be characterised. Seeds of L. linifolius were characterised using physical, colorimetric and chemical data. Ultrastructural and functional characterisation of the N-fixing root nodules included immunolabelling with nifH protein antibodies (recognising the N-fixing enzyme, nitrogenase). Endosymbiotic bacteria were isolated from root nodules and characterised phylogenetically using 16S rRNA, nodA and nodD gene sequences. L. linifolius yielded heteromorphic seed of distinct colour classes: green and brown. Seed morphotypes had similar C:N ratios and were equally germinable (ca. 90%) after scarification at differing optimal temperatures (16 and 20 °C). Brown seeds were larger and comprised a larger proportion of the seed batch (69%). L. linifolius root nodules appeared indeterminate in structure, effective (capable of fixing atmospheric N) and having strains very similar to Rhizobium leguminosarum biovar viciae. The findings and rhizobial isolates have potential application for ecological restoration and horticulture using native seeds. Also, the data and rhizobial resources have potential applications in comparative and functional studies with related and socio-economically important crops such as Pisum, Lens and Vicia.

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Analysis of rhizobial endosymbionts of Vicia, Lathyrus and Trifolium species used to maintain mountain firewalls in Sierra Nevada National Park (South Spain)

Cited by 11 publications

References 60 publications

Metabarcoding reveals that rhizospheric microbiota of Quercus pyrenaica is composed by a relatively small number of bacterial taxa highly abundant

Metabarcoding reveals that rhizospheric microbiota of Quercus pyrenaica is composed by a relatively small number of bacterial taxa highly abundant

Potential of wild annual legumes for mountain pasture restoration at two silvopastoral sites in southern Spain: promising species and soil-improvement techniques

Towards a characterisation of the wild legume bitter vetch (Lathyrus linifolius L. (Reichard) Bässler): heteromorphic seed germination, root nodule structure and N‐fixing rhizobial symbionts

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