Plant growth-promoting bacteria isolated from earthworms enhance spinach growth and its phytoremediation potential in metal-contaminated soils.

The aim of this study was to evaluate the impact of metal-tolerant plant growth-promoting bacteria (PGPB) isolated from the chloragogenous tissue of Aporrectodea molleri, which represents a unique habitat. Our objectives were to investigate their effects on the growth of Spinacia oleracea under heavy metal stress and assess their potential for enhancing phytoremediation capabilities. The experiment was conducted in an alkaline soil contaminated with 7 mg kg-1 of cadmium, 100 mg kg-1 of nickel, 150 mg kg-1 of copper, 300 mg kg-1 of Zinc, and mg kg-1 of 600 Manganese. The results showed that heavy metal stress considerably diminished root (42.8%) and shoot length (60.1%), biomass (80%), chlorophyll content (41%), soil alkaline (45%), and acid (51%) phosphatases (42%) and urease (42%). However, soil inoculation with bacterial isolates remarkably improved plant growth. Soil bioaugmentation increased spinach growth (up to 74.5% for root length, up to 106.3% for shoot length, and up to 5.5 folds for fresh biomass) while significantly increasing soil enzyme activity and NPK content. Multivariate data analysis indicated that soil inoculation with Bacillus circulans TC7 promoted plant growth while limiting metal bioaccumulation, whereas Pseudomonas sp. TC33 and Bacillus subtilis TC34 increased metal bioaccumulation in spinach tissues while minimizing their toxicity. Our study confirms that earthworms are a reservoir of multi-beneficial bacteria that can effectively improve phytoremediation efficiency and mitigate the toxic effects of heavy metals on plant growth. Further studies are needed to investigate the long-term effects and feasibility of using these isolates as a consortium in field applications.

Taxonomic and functional characteristics of aerobic bacteria isolated from the chloragogenous tissue of the earthworm Aporrectodea molleri.

Earthworms are considered as a rich microhabitat for the growth and proliferation of diverse soil microorganisms. Hence, earthworms' associated bacteria attracted interest due to their high metabolic profiles and benefits to soil fertility and plant growth. In this study, we aimed to isolate for the first-time aerobic bacteria present in the chloragogenous tissue of the earthworm Apporectodea molleri and test their Plant Growth-Promoting abilities and their resistance to heavy metals (Mn, Zn, Cu, Cd, and Ni). The 16S rRNA gene sequencing revealed the affiliation of the fifteen isolates to six main bacterial genera: Enterobacter, Citrobacter, Aeromonas, Pseudomonas, Bacillus, Terribacillus. These strains displayed different plant growth promoting traits (e.g., indole-3-acetic acid IAA, siderophores, nitrogen fixation, phosphate, and potassium solubilization), in addition, they were able to resist differently to heavy metals. Bacillus strains were most effective as three strains, namely B. subtilis strain TC34; B. circulans strain TC7 and Bacillus sp. strain TC10, were positive to all PGP traits and resisted to all heavy metals. This study illustrates the potential of bacteria from the chloragogenous tissue to exhibit multiple properties, which can be related to the functional feature of this tissue to stock metabolites and neutralize toxic elements.

Earthworms and their cutaneous excreta can modify the virulence and reproductive capability of entomopathogenic nematodes and fungi.

Earthworms are ecological engineers that can contribute to the displacement of biological control agents such as the entomopathogenic nematodes (EPNs) and fungi (EPF). However, a previous study showed that the presence of cutaneous excreta (CEx) and feeding behavior of the earthworm species Eisenia fetida (Haplotaxida: Lumbricidae) compromise the biocontrol efficacy of certain EPN species by reducing, for example, their reproductive capability. Whether this phenomenon is a general pattern for the interaction of earthworms-entomopathogens is still unknown. We hypothesized that diverse earthworm species might differentially affect EPN and EPF infectivity and reproductive capability. Here we investigated the interaction of different earthworm species (Eisenia fetida, Lumbricus terrestris, and Perionyx excavatus) (Haplotaxida) and EPN species (Steinernema feltiae, S. riojaense, and Heterorhabditis bacteriophora) (Rhabditida) or EPF species (Beauveria bassiana and Metarhizium anisopliae) (Hypocreales), in two independent experiments. First, we evaluated the application of each entomopathogen combined with earthworms or their CEx in autoclaved soil. Hereafter, we studied the impact of the earthworms' CEx on entomopathogens applied at two different concentrations in autoclaved sand. Overall, we found that the effect of earthworms on entomopathogens was species-specific. For example, E. fetida reduced the virulence of S. feltiae, resulted in neutral effects for S. riojaense, and increased H. bacteriophora virulence. However, the earthworm P. excavates increased the virulence of S. feltiae, reduced the activity of H. bacteriophora, at least at specific timings, while S. riojaense remained unaffected. Finally, none of the EPN species were affected by the presence of L. terrestris. Also, the exposure to earthworm CEx resulted in a positive, negative or neutral effect on the virulence and reproduction capability depending on the earthworm-EPN species interaction. Concerning EPF, the impact of earthworms was also differential among species. Thus, E. fetida was detrimental to M. anisopliae and B. bassiana after eight days post-exposure, whereas Lumbricus terrestris resulted only detrimental to B. bassiana. In addition, most of the CEx treatments of both earthworm species decreased B. bassiana virulence and growth. However, the EPF M. anisopliae was unaffected when exposed to L. terrestris CEx, while the exposure to E. fetida CEx produced contrasting results. We conclude that earthworms and their CEx can have positive, deleterious, or neutral impacts on entomopathogens that often coinhabit soils, and that we must consider the species specificity of these interactions for mutual uses in biological control programs. Additional studies are needed to verify these interactions under natural conditions.

Unveiling resilience: coelomic fluid bacteria's impact on plant metabolism and abiotic stress tolerance.

Earthworms' coelomic fluid (CF) has been discovered to possess properties that promote plant development. In particular, the earthworm's coelomic fluid-associated bacteria (CFB) are the primary factor influencing the plants' response. To investigate this, we used bacteria isolated from the CF and selected based on different plant growth-promoting traits, in a mesocosm ecosystem that includes plants. This experiment aimed to assess their impact on the metabolism of plants growing under abiotic stress environments (alkaline soil and nitrogen (N), phosphate (P), and potassium (K) deficit) and compare the lipid profiles of plants under the various treatments. We used seven different bacterial species isolated from the CF of Aporrectodea molleri and as a plant model Zea mays L. For the metabolomic analysis method, we used gas chromatography-mass spectrometry lipidomic. After observing the metabolomic profiles, we found that a few molecular pathways are involved in how plants react to bacterial biostimulants. The bacterial isolates belonging to Pantoea vagans, Pseudomonas aeruginosa, Bacillus paramycoides, and Bacillus thuringiensis have led to a significant increase in synthesizing several metabolites belonging to various chemical categories. Contrary to predictions, abiotic stress did not cause a drop in the composition and concentration of lipids in plants treated with the CFB, demonstrating the rigidity of the protective mechanisms. The statistical analysis based on the Pearson method revealed a positive significant correlation between plant growth parameters (length of the aerial part, surface of the leaves, and biomass) and some metabolites belonging to fatty acids, carboxylic acids, benzene derivatives, and alkanes. Moreover, the standard metabolic components of all treatments in much higher concentrations during bacterial treatments than the control treatment suggests that the bacteria have stimulated the overexpression of these metabolic components. According to these results, we could assume that plants treated with CFB exhibit an adaptability of abiotic stress defense mechanisms, which may be attributed to the upregulation of genes involved in lipid biosynthesis pathways.

Plant growth-promoting bacteria isolated from earthworms enhance spinach growth and its phytoremediation potential in metal-contaminated soils / Las bacterias promotoras del crecimiento vegetal aisladas de lombrices de tierra mejoran el crecimiento de las espinacas y su potencial de fitorremediación en suelos contaminados con metales

The aim of this study was to evaluate the impact of metal-tolerant plant growth-promoting bacteria (PGPB) isolated from the chloragogenous tissue of Aporrectodea molleri, which represents a unique habitat. Our objectives were to investigate their effects on the growth of Spinacia oleracea under heavy metal stress and assess their potential for enhancing phytoremediation capabilities. The experiment was conducted in an alkaline soil contaminated with 7 mg kg-1 of cadmium, 100 mg kg-1 of nickel, 150 mg kg-1 of copper, 300 mg kg-1 of Zinc, and mg kg-1 of 600 Manganese. The results showed that heavy metal stress considerably diminished root (42.8%) and shoot length (60.1%), biomass (80%), chlorophyll content (41%), soil alkaline (45%), and acid (51%) phosphatases (42%) and urease (42%). However, soil inoculation with bacterial isolates remarkably improved plant growth. Soil bioaugmentation increased spinach growth (up to 74.5% for root length, up to 106.3% for shoot length, and up to 5.5 folds for fresh biomass) while significantly increasing soil enzyme activity and NPK content. Multivariate data analysis indicated that soil inoculation with Bacillus circulans TC7 promoted plant growth while limiting metal bioaccumulation, whereas Pseudomonas sp. TC33 and Bacillus subtilis TC34 increased metal bioaccumulation in spinach tissues while minimizing their toxicity. Our study confirms that earthworms are a reservoir of multi-beneficial bacteria that can effectively improve phytoremediation efficiency and mitigate the toxic effects of heavy metals on plant growth. Further studies are needed to investigate the long-term effects and feasibility of using these isolates as a consortium in field applications.(AU)