Luciane Maria Pereira Passaglia

Plant growth-promoting bacteria as inoculants in agricultural soils

Abstract Plant-microbe interactions in the rhizosphere are the determinants of plant health, productivity and soil fertility. Plant growth-promoting bacteria (PGPB) are bacteria that can enhance plant growth and protect plants from disease and abiotic stresses through a wide variety of mechanisms; those that establish close associations with plants, such as the endophytes, could be more successful in plant growth promotion. Several important bacterial characteristics, such as biological nitrogen fixation, phosphate solubilization, ACC deaminase activity, and production of siderophores and phytohormones, can be assessed as plant growth promotion (PGP) traits. Bacterial inoculants can contribute to increase agronomic efficiency by reducing production costs and environmental pollution, once the use of chemical fertilizers can be reduced or eliminated if the inoculants are efficient. For bacterial inoculants to obtain success in improving plant growth and productivity, several processes involved can influence the efficiency of inoculation, as for example the exudation by plant roots, the bacterial colonization in the roots, and soil health. This review presents an overview of the importance of soil-plant-microbe interactions to the development of efficient inoculants, once PGPB are extensively studied microorganisms, representing a very diverse group of easily accessible beneficial bacteria.

Genetic and phenotypic diversity of plant-growth-promoting bacilli isolated from wheat fields in southern Brazil.

In this work, a total of 311 putative nitrogen-fixing bacilli were isolated from seven distinct wheat production zones of the Rio Grande do Sul State, Brazil. Strains belonging to several species were grouped into 40 different nifH-RFLP-PCR profiles. The genus Paenibacillus was the most prominent group in both the rhizosphere (77.8%) and soil (79%). Paenibacillus borealis was the most frequently identified species, followed by Paenibacillus graminis. The remainder of the isolated bacteria belonged to the genus Bacillus sp. Indolic compound production (indole 3-acetic acid (IAA), indolepyruvic acid (IPyA) and indoleacetamide (IAM)) was detected in 33.6% and 26% of the isolates from the rhizosphere and soil, respectively. Among the 311 isolates, nine were able to solubilize phosphate and 48 were able to produce siderophores. The isolates SBR5, CSR16 and EsR7, identified by the 16S rRNA gene sequence as strains of Paenibacillus sp., were chosen for in vivo experiments in a greenhouse and proved to be very efficient in promoting a significant increase in the shoot and dry matter of wheat plants. Those strains could be useful in formulation of new inoculants, improving the cropping systems into which they can be most profitably applied.

The effect of plant growth-promoting rhizobacteria on the growth of rice (Oryza sativa L.) cropped in southern Brazilian fields

Background and AimsSeveral strains of rhizobacteria may be found in the rhizospheric soil, on the root surface or in association with rice plants. These bacteria are able to colonize plant root systems and promote plant growth and crop yield through a variety of mechanisms. The objectives of this study were to isolate, identify, and characterize putative plant growth-promoting rhizobacteria (PGPR) associated with rice cropped in different areas of southern Brazil.MethodsBacterial strains were selectively isolated based on their growth on three selective semi-solid nitrogen-free media. Bacteria were identified at the genus level by PCR-RFLP 16S rRNA gene analysis and partial sequencing methodologies. Bacterial isolates were evaluated for their ability to produce indolic compounds and siderophores and to solubilize phosphate. In vitro biological nitrogen fixation and the ability to produce 1-aminocyclopropane-1-carboxylate deaminase were evaluated for each bacterial isolate used in the inoculation experiments.ResultsIn total, 336 bacterial strains were isolated representing 31 different bacterial genera. Strains belonging to the genera Agrobacterium, Burkholderia, Enterobacter, and Pseudomonas were the most prominent isolates. Siderophore and indolic compounds producers were widely found among isolates, but 101 isolates were able to solubilize phosphate. Under gnotobiotic conditions, eight isolates were able to stimulate the growth of rice plants. Five of these eight isolates were also field tested in rice plants subjected to different nitrogen fertilization rates.ConclusionsThe results showed that the condition of half-fertilization plus separate inoculation with the isolates AC32 (Herbaspirillum sp.), AG15 (Burkholderia sp.), CA21 (Pseudacidovorax sp.), and UR51 (Azospirillum sp.) achieved rice growth similar to those achieved by full-fertilization without inoculation, thus highlighting the potential of these strains for formulating new bioinoculants for rice crops.

Screening of plant growth promoting Rhizobacteria isolated from sunflower (Helianthus annuus L.)

Background and AimsThis study was aimed at assessing the diversity of putatively diazotrophic rhizobacteria associated with sunflower (Helianthus annuus L.) cropped in the south of Brazil, and to examine key plant growth promotion (PGP) characteristics of the isolates for the purposes of increasing plant productivity.Methods299 strains were isolated from the roots and rhizosphere of sunflower cultivated in five different areas using N-free media. 16S rDNA PCR-RFLP and 16S rRNA partial sequencing were used for identification and the Shannon index was used to evaluate bacterial diversity. Production of siderophores and indolic compounds (ICs), as well phosphate solubilization activities of each isolate were also evaluated in vitro. On the basis of multiple PGP activities, eight isolates were selected and tested for their N-fixation ability, and their capacity as potential PGPR on sunflower plants was also assessed.ResultsAll except three Gram-positive strains (phylum Actinobacteria) belonged to the Gram-negative Proteobacteria subgroups [Gamma (167), Beta (78), and Alpha (50)] and the family Flavobacteriaceae (1)]. Shannon indexes ranged from 0.96 to 2.13 between the five sampling sites. Enterobacter and Burkholderia were the predominant genera isolated from roots and rhizosphere, respectively. Producers of siderophores and ICs were widely found amongst the isolates, but only 19.8% of them solubilized phosphate. About 8% of the isolates exhibited all three PGP traits, and these mostly belonged to the genus Burkholderia. Four isolates were able to stimulate the growth of sunflower plants under gnotobiotic conditions.ConclusionsEnterobacter and Burkholderia were the dominant rhizospheric bacterial genera associated with sunflower plants. Inoculation with isolates belonging to the genera Achromobacter, Chryseobacterium, Azospirillum, and Burkholderia had a stimulatory effect on plant growth.

Paenibacillus riograndensis sp. nov., a nitrogen- fixing species isolated from the rhizosphere of Triticum aestivum

A bacterial strain designated SBR5(T) was isolated from the rhizosphere of Triticum aestivum. A phylogenetic analysis based on the 16S rRNA gene sequence placed the isolate within the genus Paenibacillus, being most closely related to Paenibacillus graminis RSA19(T) (98.1 % similarity). The isolate was a Gram-reaction-variable, motile, facultatively anaerobic bacterium, with spores in a terminal position in cells. Starch was utilized and dihydroxyacetone and catalase were produced. Strain SBR5(T) displayed plant-growth-promoting rhizobacteria characteristics: the ability to fix nitrogen and to produce siderophores and indole-3-acetic acid. The DNA G+C content was 55.1 mol%. Chemotaxonomic analysis of the isolated strain revealed that MK-7 was the predominant menaquinone, while the major fatty acid was anteiso-C(15 : 0). DNA-DNA hybridization values between strain SBR5(T) and P. graminis RSA19(T), Paenibacillus odorifer TOD45(T) and Paenibacillus borealis KK19(T) were 43, 35 and 28 %, respectively. These DNA relatedness data and the results of phylogenetic and phenotypic analyses showed that strain SBR5(T) should be considered as the nitrogen-fixing type strain of a novel species of the genus Paenibacillus, for which the name Paenibacillus riograndensis sp. nov. is proposed. The type strain is SBR5(T) (=CCGB 1313(T) =CECT 7330(T)).

The effects of different fertilization conditions on bacterial plant growth promoting traits: guidelines for directed bacterial prospection and testing

AimsIn this work, the effects of fertilization on diazotroph diversity and plant growth promoting traits were analyzed. An interaction model was then suggested and tested in field.MethodsOne hundred and ninety bacterial strains were isolated from rhizospheric soil and roots of rice cropped in three different fertilization conditions. Phosphate solubilization, indolic compound (IC) and siderophore production, and nitrogen fixation abilities of the isolates were screened and compared. Some isolates were selected for in vivo plant growth promotion in greenhouse and field experiments.ResultsWe found that fertilization had a limited effect on diversity but had a major effect on phosphate solubilization and IC production abilities. We demonstrated that plants select bacteria that present good phosphate solubilization ability for intimate root association in lieu of good IC production under nutrient-poor conditions and select good IC producers in lieu of good phosphate solubilizers under nutrient-moderate conditions. In nutrient-rich conditions, this selection preference seems to be deactivated. In the field trial, good phosphate solubilizers only contributed effectively to plant growth at nutrient-poor conditions and good IC producers only contributed to plant growth at nutrient-moderate conditions.ConclusionsFertilization affects the PGP traits of the diazotrophic community. These findings may be used for directed PGPR prospection and anticipated PGPR candidate selection.

Occurrence of plant growth-promoting traits in clover-nodulating rhizobia strains isolated from different soils in Rio Grande do Sul state

In the last decades, the use of plant growth-promoting rhizobacteria has become an alternative to improve crop production. Rhizobium leguminosarum biovar trifolii is one of the most promising rhizobacteria and is even used with non-legume plants. This study investigated in vitro the occurrence of plant growth-promoting characteristics in several indigenous R. leguminosarum biovar trifolii isolated from soils in the State of Rio Grande do Sul, Brazil. Isolates were obtained at 11 locations and evaluated for indoleacetic acid and siderophore production and inorganic phosphate solubilization. Ten isolates were also molecularly characterized and tested for antagonism against a phytopathogenic fungus and for plant growth promotion of rice seedlings. Of a total of 252 isolates, 59 produced indoleacetic acid, 20 produced siderophores and 107 solubilized phosphate. Some degree of antagonism against Verticillium sp. was observed in all tested isolates, reducing mycelial growth in culture broth. Isolate AGR-3 stood out for increasing root length of rice seedlings, while isolate ELD-18, besides increasing root length in comparison to the uninoculated control, also increased the germination speed index, shoot length, and seedling dry weight. These results confirm the potential of some strains of R. leguminosarum biovar trifolii as plant growth-promoting rhizobacteria.

Ecological role of bacterial inoculants and their potential impact on soil microbial diversity

BackgroundMicrobial inoculants are an alternative method of increasing crop productivity that can reduce the use of chemical fertilizers, which is one of the more controversial agricultural practices that affect the environment. Beneficial bacteria, collectively known as plant growth-promoting bacteria (PGPB), enhance plant growth and protect plants from disease and abiotic stresses through a wide variety of mechanisms. Bacterial inoculation efficiency is associated with the beneficial features of the inoculated bacterium, as well as with the complex network of interactions occurring in the soil.ScopeBeneficial bacteria have previously been examined for interactions with different plant hosts, soil types, and agricultural practices, but there is limited information concerning the potential effects of the release of microorganisms on soil functionality. Despite the plant growth promotion characteristics, the survival, abundance, and persistence of inoculant in soil or plant roots are characteristics that could potentially lead to its invasiveness. Inoculants can also interfere with soil health and microbial and faunal community composition.ConclusionThis review presents an overview of plant-PGPB interactions and their impacts on microbial communities, hypothesizing about the potential of these interactions to promote positive disturbances in soil, mainly in poor environments. The inoculation of free-living bacteria seems to cause a short-term impact to agricultural soils, while rhizobia-based inoculants or bacterial inoculations performed under stress conditions are long-term processes. However, there is great variability amongst results concerning the effects of bacterial inoculation into different plant and soil conditions.

A Model to Explain Plant Growth Promotion Traits: A Multivariate Analysis of 2,211 Bacterial Isolates

Plant growth-promoting bacteria can greatly assist sustainable farming by improving plant health and biomass while reducing fertilizer use. The plant-microorganism-environment interaction is an open and complex system, and despite the active research in the area, patterns in root ecology are elusive. Here, we simultaneously analyzed the plant growth-promoting bacteria datasets from seven independent studies that shared a methodology for bioprospection and phenotype screening. The soil richness of the isolates origin was classified by a Principal Component Analysis. A Categorical Principal Component Analysis was used to classify the soil richness according to isolates indolic compound production, siderophores production and phosphate solubilization abilities, and bacterial genera composition. Multiple patterns and relationships were found and verified with nonparametric hypothesis testing. Including niche colonization in the analysis, we proposed a model to explain the expression of bacterial plant growth-promoting traits according to the soil nutritional status. Our model shows that plants favor interaction with growth hormone producers under rich nutrient conditions but favor nutrient solubilizers under poor conditions. We also performed several comparisons among the different genera, highlighting interesting ecological interactions and limitations. Our model could be used to direct plant growth-promoting bacteria bioprospection and metagenomic sampling.

Changes in root bacterial communities associated to two different development stages of canola (Brassica napus L. var oleifera) evaluated through next-generation sequencing technology.

Crop production may benefit from plant growth-promoting bacteria. The knowledge on bacterial communities is indispensable in agricultural systems that intend to apply beneficial bacteria to improve plant health and production of crops such as canola. In this work, the diversity of root bacterial communities associated to two different developmental phases of canola (Brassica napus L.) plants was assessed through the application of new generation sequencing technology. Total bacterial DNA was extracted from root samples from two different growth states of canola (rosette and flowering). It could be shown how bacterial communities inside the roots changed with the growing stage of the canola plants. There were differences in the abundance of the genera, family, and even the phyla identified for each sample. While in both root samples Proteobacteria was the most common phylum, at the rosette stage, the most common bacteria belonged to the family Pseudomonadaceae and the genus Pseudomonas, and in the flowering stage, the Xanthomonadaceae family and the genus Xanthomonas dominated the community. This implies in a switch in the predominant bacteria in the different developmental stages of the plant, suggesting that the plant itself interferes with the associated microbial community.