The inoculation with Ensifer meliloti sv. rigiduloides improves considerably the growth of Robinia pseudoacacia under lead-stress

Aims: In Morocco, the Zaida abandoned mine is a major source of metallic trace elements that are harmful to the environment and human health. With the aim to use the heavy metal tolerant legume tree Robinia pseudoacacia in the phytostabilization of this mine’s tailings, we isolated and characterized its indigenous microsymbionts, and tested their effects on the plant growth under high Pb-acetate concentrations. Methods: Thus, 27 bacteria were isolated from nodules of the plant grown in the Zaida abandoned mine tailings, and we examined their tolerance to high concentrations of Pb-acetate in-vitro. Four strains were then selected for symbiotic, molecular, and phenotypic analyses, and we analyzed the effect of inoculation on the plant growth parameters under increasing Pb-acetate concentrations (in-vivo). Results: The isolated strains tolerate high lead concentrations. The Phylogenetic analyses of 16 S rRNA, five housekeeping genes (MLSA), and three symbiotic genes showed the affiliation of the strains with Ensifer meliloti sv. rigiduloides and E. kummerowiae sv. meliloti. Representative strains of both symbiovars tolerate 1100 µg ml Pb-acetate in-vitro and nodulate R. pseudoacacia even under 1200 µg ml of Pb-acetate. In all tests, the plant proline content increased as a response mechanism to increasing concentrations of Pb-acetate. Inoculation experiments under high concentrations of Pb-acetate showed that both strains improved the plant growth parameters and chlorophyll content compared to controls. Conclusion: Our results show that E. meliloti sv. rigiduloides strain RPZ12 improves plant growth under Pb stress conditions more than E. kummerowiae sv. meliloti strain RPZ17. Strain RPZ12 would be an excellent inoculum to use for the phytostabilization of Zaida mine tailings. This is the first description of symbiovar meliloti in E. kummerowiae.The authors want to thank all the persons who contributed to the achievement of this work. This work was supported by the Moroccan ministry of higher education. Dr Alami received a grant from the ministr

Plant growth promoting activities of Pseudomonas sp. and Enterobacter sp. isolated from the rhizosphere of Vachellia gummifera in Morocco

The Moroccan endemic Vachellia gummifera grows wild under extreme desert conditions. This plant could be used as an alternative fodder for goats, and camels, in order to protect the Argan forests against overgrazing in Central and Southwestern Moroccan semiarid areas. With the aim to improve the V. gummifera population's density in semiarid areas, we proposed its inoculation with performing plant growth-promoting bacteria. Hence, 500 bacteria were isolated from the plant rhizosphere. From these, 291 isolates were retained for plant growth-promoting (PGP) activities assessment. A total of 44 isolates showed the best phosphates solubilization potential, as well as siderophore and auxin production. The combination of REP-PCR (repetitive extragenic palindromic-polymerase chain reaction) fingerprinting, PGP activities, and phenotypic properties, allowed the selection of three strains for the inoculation experiments. The three selected strains' 16S rRNA sequencing showed that they are members of the Enterobacter and Pseudomonas genera. The inoculation with three strains had diverse effects on V. gummifera growth parameters. All single and combined inoculations improved the plant shoot weight by more than 200%, and the root length by up to 139%, while some combinations further improved protein and chlorophyll content, thereby improving the plant's forage value. The three selected strains constitute an effective inoculum for use in the arid and semiarid zones of southern Morocco.The authors want to thank all the persons who contributed to the achievement of this work. This work was funded by the Hassan II Academy of Science and Technology (AH2ST), in Morocco. B.M. received a grant from the AH2ST. She received also the excellency fellowship Khadija Baddouri in life sciences, from the Horizons foundation, Franc

Different species of Bradyrhizobium from symbiovars genistearum and retamae nodulate the endemic Retama dasycarpa in the High Atlas Mountains

Retama dasycarpa is an endemic Retama species native to the cold semi-arid bioclimates of the High Atlas Mountains in Morocco. In this work, we analyzed the diversity of the microsymbionts nodulating this plant and their different phenotypic and symbiotic characteristics. Phylogenetic analysis of the 16S rRNA gene revealed that the tested isolates clustered in the Bradyrhizobium genus. Multilocus sequence analyses of four housekeeping genes (recA, gyrB, glnII and atpD) for 12 selected strains grouped them into four clusters close to B. lupini USDA 3051, B. frederickii CNPSo 3446, B. valentinum LmjM3 and B. retamae Ro19. The individual phylogenies of these core genes and the symbiotic genes nodC, nodA and nifH were congruent. These isolates showed a broad host range, being able to nodulate different legume hosts, such as R. sphaerocarpa, R. monosperma, Lupinus luteus, Cytisus grandiflorus and Chamaecytisus albidus, but not Phaseolus vulgaris or Glycine max. They all had a similar metabolic capacity, using the majority of the carbohydrates and amino acids tested as sole sources of carbon and nitrogen. Furthermore, out of the 12 selected strains, some displayed plant growth-promoting features, with six of them solubilizing phosphate and three of them producing siderophores. The present work provides, for the first time, a detailed description about the microsymbionts associated with the endemic legume R. dasycarpa.This work was supported by the Ministry of Higher Education and Innovation. Dr Mouad Lamrabet was granted a fellowship from the PPR2-BIOMIVER project. The authors want to thank all the people who contributed to this work

Different species of Bradyrhizobium from Symbiovars Genistearum and Retamae Nodulate the endemic Retama dasycarpa in the High Atlas Mountains

Resumen del poster presntado en: 15th European Nitrogen Fixation Conference (ENFC). Napóles Italia. 31 agosto - 3 septiembre (2023