Effect of Co-Inoculation with Growth-Promoting Bacteria and Arbuscular Mycorrhizae on Growth of Persea americana Seedlings Infected with Phytophthora cinnamomi.

Avocado is one of the most in-demand fruits worldwide and the trend towards its sustainable production, regulated by international standards, is increasing. One of the most economically important diseases is root rot, caused by Phythopthora cinnamomi. Regarding this problem, antagonistic microorganism use is an interesting alternative due to their phytopathogen control efficiency. Therefore, the interaction of arbuscular mycorrhizal fungi of the phylum Glomeromycota, native to the Peruvian coast (GWI) and jungle (GFI), and avocado rhizospheric bacteria, Bacillus subtilis and Pseudomonas putida, was evaluated in terms of their biocontrol capacity against P. cinnamomi in the "Zutano" variety of avocado plants. The results showed that the GWI and Bacillus subtilis combination increased the root exploration surface by 466.36%. P. putida increased aerial biomass by 360.44% and B. subtilis increased root biomass by 433.85%. Likewise, P. putida rhizobacteria showed the highest nitrogen (24.60 mg ∙ g-1 DM) and sulfur (2.60 mg ∙ g-1 DM) concentrations at a foliar level. The combination of GWI and Bacillus subtilis was the treatment that presented the highest calcium (16.00 mg ∙ g-1 DM) and magnesium (8.80 mg ∙ g-1 DM) concentrations. The microorganisms' multifunctionality reduced disease severity by 85 to 90% due to the interaction between mycorrhizae and rhizobacteria. In conclusion, the use of growth promoting microorganisms that are antagonistic to P. cinnamomi represents a potential strategy for sustainable management of avocado cultivation.

Role of Rahnella aquatilis AZO16M2 in Phosphate Solubilization and Ex Vitro Acclimatization of Musa acuminata var. Valery.

Rahnella aquatilis AZO16M2, was characterized for its phosphate solubilization capacity to improve the establishment and survival of Musa acuminata var. Valery seedlings under ex-acclimation. Three phosphorus sources (Rock Phosphate (RF), Ca3(PO4)2 and K2HPO4) and two types of substrate (sand:vermiculite (1:1) and Premix N°8) were selected. The factorial analysis of variance (p < 0.05) showed that R. aquatilis AZO16M2 (OQ256130) solubilizes Ca3(PO4)2 in solid medium, with a Solubilization Index (SI) of 3.77 at 28 °C (pH 6.8). In liquid medium, it was observed that R. aquatilis produced 29.6 mg/L soluble P (pH 4.4), and synthesized organic acids (oxalic, D-gluconic, 2-ketogluconic and malic), Indole Acetic Acid (IAA) (33.90 ppm) and siderophores (+). Additionally, acid and alkaline phosphatases (2.59 and 2.56 µg pNP/mL/min) were detected. The presence of the pyrroloquinoline-quinone (PQQ) cofactor gene was confirmed. After inoculating AZO16M2 to M. acuminata in sand:vermiculite with RF, the chlorophyll content was 42.38 SPAD (Soil Plant Analysis Development). Aerial fresh weight (AFW), aerial dry weight (ADW) and root dry weight (RDW) were superior to the control by 64.15%, 60.53% and 43.48%, respectively. In Premix N°8 with RF and R. aquatilis, 8.91% longer roots were obtained, with 35.58% and 18.76% more AFW and RFW compared with the control as well as 94.45 SPAD. With Ca3(PO4)2, values exceeded the control by 14.15% RFW, with 45.45 SPAD. Rahnella aquatilis AZO16M2 favored the ex-climatization of M. acuminata through improving seedling establishment and survival.

Plant Growth Promoting Bacteria and Arbuscular Mycorrhizae Improve the Growth of Persea americana var. Zutano under Salt Stress Conditions.

In Peru, almost 50% of the national agricultural products come from the coast, highlighting the production of avocado. Much of this area has saline soils. Beneficial microorganisms can favorably contribute to mitigating the effect of salinity on crops. Two trials were carried out with var. Zutano to evaluate the role of native rhizobacteria and two Glomeromycota fungi, one from a fallow (GFI) and the other from a saline soil (GWI), in mitigating salinity in avocado: (i) the effect of plant growth promoting rhizobacteria, and (ii) the effect of inoculation with mycorrhizal fungi on salt stress tolerance. Rhizobacteria P. plecoglissicida, and B. subtilis contributed to decrease the accumulation of chlorine, potassium and sodium in roots, compared to the uninoculated control, while contributing to the accumulation of potassium in the leaves. Mycorrhizae increased the accumulation of sodium, potassium, and chlorine ions in the leaves at a low saline level. GWI decreased the accumulation of sodium in the leaves compared to the control (1.5 g NaCl without mycorrhizae) and was more efficient than GFI in increasing the accumulation of potassium in leaves and reducing chlorine root accumulation. The beneficial microorganisms tested are promising in the mitigation of salt stress in avocado.

Bradyrhizobium centrosemae (symbiovar centrosemae) sp. nov., Bradyrhizobium americanum (symbiovar phaseolarum) sp. nov. and a new symbiovar (tropici) of Bradyrhizobium viridifuturi establish symbiosis with Centrosema species native to America.

In this work we analyze through a polyphasic approach several Bradyrhizobium strains isolated in Venezuela from root nodules of Centrosema species. The analysis of the 16S rRNA gene showed that the strains belong to three clusters within genus Bradyrhizobium which have 100% similarity with Bradyrhizobium daqingense CCBAU 15774(T)Bradyrhizobium guangxiense CCBAU 53363(T) and Bradyrhizobium viridifuturi SEMIA 690(T). The results of recA and glnII gene analysis confirmed the identification of the strains CMVU02 and CMVU30 as Bradyrhizobium viridifuturi but the nodC gene analysis showed that they belong to a new symbiovar for which we propose the name tropici. Nevertheless, the concatenated recA and glnII gene phylogenetic analysis, DNA-DNA hybridization and phenotypic characterization showed that the strains A9(T), CMVU44(T) and CMVU04 belong to two novel Bradyrhizobium species. The analysis of the nodC gene showed that these strains also represent two new symbiovars. Based on these results we propose the classification of the strain A9(T) isolated from Centrosema molle into the novel species Bradyrhizobium centrosemae (sv. centrosemae) sp. nov. (type strain A9(T)=LMG 29515(T)=CECT 9095(T)). and the classification of the strains CMVU44(T) and CMVU04 isolated from C. macrocarpum into the novel species Bradyrhizobium americanum (sv. phaseolarum) sp. nov. (type strain CMVU44(T)=LMG 29514(T)=CECT 9096(T)).

Pseudomonas guariconensis sp. nov., isolated from rhizospheric soil.

We isolated a bacterial strain designated PCAVU11(T) in the course of a study of phosphate-solubilizing bacteria occurring in rhizospheric soil of Vigna unguiculata (L.) Walp. in Guárico state, Venezuela. The 16S rRNA gene sequence had 99.2 % sequence similarity with respect to the most closely related species, Pseudomonas taiwanensis, and 99.1 % with respect to Pseudomonas entomophila, Pseudomonas plecoglossicida and Pseudomonas monteilii, on the basis of which PCAVU11(T) was classified as representing a member of the genus Pseudomonas. Analysis of the housekeeping genes rpoB, rpoD and gyrB confirmed the phylogenetic affiliation and showed sequence similarities lower than 95 % in all cases with respect to the above-mentioned closest relatives. Strain PCAVU11(T) showed two polar flagella. The respiratory quinone was Q9. The major fatty acids were 16 : 0 (25.7 %), 18 : 1ω7c (20.4 %), 17 : 0 cyclo (11.5 %) and 16 : 1ω7c/15 : 0 iso 2-OH in summed feature 3 (10.8 %). The strain was oxidase-, catalase- and urease-positive, the arginine dihydrolase system was present but nitrate reduction, ß-galactosidase production and aesculin hydrolysis were negative. Strain PCAVU11(T) grew at 44 °C and at pH 10. The DNA G+C content was 61.5 mol%. DNA-DNA hybridization results showed values lower than 56 % relatedness with respect to the type strains of the four most closely related species. Therefore, the results of genotypic, phenotypic and chemotaxonomic analyses support the classification of strain PCAVU11(T) as representing a novel species of the genus Pseudomonas, which we propose to name Pseudomonas guariconensis sp. nov. The type strain is PCAVU11(T) ( = LMG 27394(T) = CECT 8262(T)).

Centrosema is a promiscuous legume nodulated by several new putative species and symbiovars of Bradyrhizobium in various American countries.

Centrosema is an American indigenous legume that can be used in agroecosystems for recovery of acidic and degraded soils. In this study, a Centrosema-nodulating rhizobial collection of strains isolated in a poor acid savanna soil from Venezuela was characterized, and the members of the collection were compared to other Centrosema strains from America. The analysis of the rrs gene showed that the strains nodulating Centrosema in American countries were closely related to different species of the genus Bradyrhizobium. However, the analysis of the atpD and recA genes, as well as the 16S-23S ITS region, showed that they formed several new phylogenetic lineages within this genus. The Venezuela strains formed three lineages that were divergent among themselves and with respect to those formed by Centrosema strains isolated in other countries, as well as to the currently described species and genospecies of Bradyrhizobium. In addition, the symbiotic genes nodC and nifH carried by Centrosema-nodulating strains were analyzed for the first time, and it was shown that they belonged to three new phylogenetic lineages within Bradyrhizobium. The nodC genes of the Centrosema strains were divergent among themselves and with respect to the genistearum and glycinearum symbiovars, indicating that Centrosema is a promiscuous legume. According to these results, the currently known Centrosema-nodulating strains represent several new putative species and symbiovars of the genus Bradyrhizobium.