The use of transgenic canola (Brassica napus) and plant growth-promoting bacteria to enhance plant biomass at a nickel-contaminated field site

The applicability of transgenic plants and plant growth-promoting bacteria to improve plant biomass accumulation as a phytoremediation strategy at a nickel (Ni)-contaminated field site was examined. Two crops of 4-day old non-transformed and transgenic canola (Brassica napus) seedlings in the presence and absence of Pseudomonas putida strain UW4 (crop #1) or P. putida strain HS-2 (crop #1 and 2) were transplanted at a Ni-contaminated field site in 2005. Overall, transgenic canola had increased growth but decreased shoot Ni concentrations compared to non-transformed canola, resulting in similar total Ni per plant. Under optimal growth conditions (crop #2), the addition of P. putida HS-2 significantly enhanced growth for non-transformed canola. Canola with P. putida HS-2 had trends of higher total Ni per plant than canola without P. putida HS-2, indicating the potential usefulness of this bacterium in phytoremediation strategies. Modifications to the planting methods may be required to increase plant Ni uptake.     

Plant Growth-Promoting Bacteria Facilitate the Growth of Barley and Oats in Salt-Impacted Soil: Implications for Phytoremediation of Saline Soils

Plant growth-promoting bacteria (PGPB) strains that contain the enzyme 1-amino- cyclopropane-1-carboxylate (ACC) deaminase can lower stress ethylene levels and improve plant growth. In this study, ACC deaminase-producing bacteria were isolated from a salt-impacted (～50 dS/m) farm field, and their ability to promote plant growth of barley and oats in saline soil was investigated in pouch assays (1% NaCl), greenhouse trials (9.4 dS/m), and field trials (6–24 dS/m). A mix of previously isolated PGPB strains UW3 (Pseudomonas sp.) and UW4 (P. sp.) was also tested for comparison. Rhizobacterial isolate CMH3 (P. corrugata) and UW3+UW4 partially alleviated plant salt stress in growth pouch assays. In greenhouse trials, CMH3 enhanced root biomass of barley and oats by 200% and 50%, respectively. UW3+UW4, CMH3 and isolate CMH2 also enhanced barley and oat shoot growth by 100%–150%. In field tests, shoot biomass of oats tripled when treated with UW3+UW4 and doubled with CHM3 compared with that of untreated plants. PGPB treatment did not affect salt uptake on a per mass basis; higher plant biomass led to greater salt uptake, resulting in decreased soil salinity. This study demonstrates a method for improving plant growth in marginal saline soils. Associated implications for salt remediation are discussed.     

Antimicrobial activities of commercial nanoparticles against an environmental soil microbe, Pseudomonas putida KT2440

Background  

The release of heavy metal-containing nanoparticles (NP) into the environment may be harmful to the efficacy of beneficial microbes that function in element cycling, pollutant degradation and plant growth. Nanoparticles of Ag, CuO and ZnO are of interest as antimicrobials against pathogenic bacteria. We demonstrate here their antimicrobial activity against the beneficial soil microbe, Pseudomonas putida KT2440.     

Evidence for the involvement of ACC deaminase from Pseudomonas putida UW4 in the biocontrol of pine wilt disease caused by Bursaphelenchus xylophilus

Pine wilt disease, caused by the nematode Bursaphelenchus xylophilus, is responsible for devastation of pine forests worldwide. Until now, there are no effective ways of dealing with this serious threat. The use of 1-aminocyclopropane-1-carboxylate (ACC) deaminase (encoded by the acdS gene)-producing plant growth-promoting bacteria has been shown to be a useful strategy to reduce the damage due to biotic and abiotic stresses. Pinus pinaster seedlings inoculated with the ACC deaminase-producing bacterium Pseudomonas putida strain UW4 showed an increased root and shoot development and reduction of B. xylophilus induced symptoms. In contrast, a P. putida UW4 acdS mutant was unable to promote pine seedling growth or to decrease B. xylophilus induced symptoms. This is the first report on the use of ACC deaminase-producing bacteria as a potential biological control agent for a tree disease, thus suggesting that the inoculation of pine seedlings grown in a tree nursery might constitute a novel strategy to obtain B. xylophilus resistant pine trees.     

The impact of Pseudomonas putida UW3 and UW4 strains on photosynthetic activities of selected field crops under saline conditions

Abstract

This research was aimed to assess the photosynthetic activities of barley (Hordeum valgare L.), clover (Trifolium repens L.), and pearl millet (Pennisetum glaucum (L.) R. Br.) under different saline conditions with two strains of Pseudomonas putida (UW3 and UW4) treatments. An exceptional observation was revealed on barley biomass ratio (288.8%) that irrigated with brackish saline water (10,000?mg/L) with the presence of P. putida UW4 strain. In general, P. putida UW3 strain was significantly increased crops biomass ratio (249.4%, 202.1%, and 212.5%) for barley, pearl millet, and clover, respectively, which were irrigated with 10,000?mg/L brackish saline water. Plant root and shoot systems were significantly increased in their length and weight reflecting the improvement of plants’ photosynthetic activities under salt stress conditions with the presence of P. putida strains. The results from pulse amplitude modulation fluorometry showed that the plants were recovered from the saline stress effect once P. putida strains were applied. The outcome of this research was highly recommended to apply P. putida strains (UW3 and UW4) with field crops for phytoremediation, in particular, where salinity (soil and/or brackish water) was environmentally challenging.     

Interactions between Pseudomonas putida UW4 and Gigaspora rosea BEG9 and their consequences for the growth of cucumber under salt-stress conditions

Aims: After the determination of the toxic but nonlethal concentration of NaCl for cucumber, we examined the interaction between an ACC (1‐aminocyclopropane‐1‐carboxylate) deaminase producing bacterial strain and an arbuscular mycorrhizal fungus (AMF) and their effects on cucumber growth under salinity. Methods and Results: In the first experiment, cucumber seedlings were exposed to 0·1, 50, 100 or 200 mmol l^?1 NaCl, and plant biomass and leaf area were measured. While seeds exposed to 200 mmol l^?1 NaCl did not germinate, plant growth and leaf size were reduced by 50 or 100 mmol l^?1 salt. The latter salt cancentration caused plant death in 1 month. In the second experiment, seeds were inoculated with the ACC deaminase‐producing strain Pseudomonas putida UW4 (AcdS⁺), its mutant unable to produce the enzyme (AcdS^?), or the AMF Gigaspora rosea BEG9, individually or in combination and exposed to 75 mmol l^?1 salt. Plant morphometric and root architectural parameters, mycorrhizal and bacterial colonization and the influence of each micro‐organism on the photosynthetic efficiency were evaluated. The AcdS⁺ strain or the AMF, inoculated alone, increased plant growth, affected root architecture and improved photosynthetic activity. Mycorrhizal colonization was inhibited by each bacterial strain. Conclusions: Salinity negatively affects cucumber growth and health, but root colonization by ACC deaminase‐producing bacteria or arbuscular mycorrhizal fungi can improve plant tolerance to such stressful condition. Significance and Impact of the Study: Arbuscular mycorrhizal fungus and bacterial ACC deaminase may ameliorate plant growth under stressful conditions. It was previously shown that, under optimal growth conditions, Ps. putida UW4 AcdS⁺ increases root colonization by Gi. rosea resulting in synergistic effects on cucumber growth. These results suggest that while in optimal conditions ACC deaminase is mainly involved in the bacteria/fungus interactions, while under stressful conditions this enzyme plays a role in plant/bacterium interactions. This finding is relevant from an ecological and an applicative point of view.     

Plant growth-promoting activities of fluorescent pseudomonads,isolated from the Iranian soils

Fluorescent pseudomonads are among the most influencing plant growth-promoting rhizobacteria in plants rhizosphere. In this research work the plant growth-promoting activities of 40 different strains of Pseudomonas fluorescens and Pseudomonas putida, previously isolated from the rhizosphere of wheat (Triticum aestivum L.) and canola (Brassica napus L.) and maintained in the microbial collection of Soil and Water Research Institute, Tehran, Iran, were evaluated. The ability of bacteria to produce auxin and siderophores and utilizing P sources with little solubility was determined. Four strains of Wp1 (P. putida), Cfp10 (Pseudomonas sp.), Wp150 (P. putida), and Wp159 (P. putida) were able to grow in the DF medium with ACC. Thirty percent of bacterial isolates from canola rhizosphere and 33% of bacterial isolates from wheat rhizosphere were able to produce HCN. The results indicate that most of the bacteria, tested in the experiment, have plant growth-promoting activities. This is the first time that such PGPR species are isolated from the Iranian soils. With respect to their great biological capacities they can be used for wheat and canola inoculation in different parts of the world, which is of very important agricultural implications.     

The Complete Genome Sequence of the Plant Growth-Promoting Bacterium Pseudomonas sp. UW4

The plant growth-promoting bacterium (PGPB) Pseudomonas sp. UW4, previously isolated from the rhizosphere of common reeds growing on the campus of the University of Waterloo, promotes plant growth in the presence of different environmental stresses, such as flooding, high concentrations of salt, cold, heavy metals, drought and phytopathogens. In this work, the genome sequence of UW4 was obtained by pyrosequencing and the gaps between the contigs were closed by directed PCR. The P. sp. UW4 genome contains a single circular chromosome that is 6,183,388 bp with a 60.05% G+C content. The bacterial genome contains 5,423 predicted protein-coding sequences that occupy 87.2% of the genome. Nineteen genomic islands (GIs) were predicted and thirty one complete putative insertion sequences were identified. Genes potentially involved in plant growth promotion such as indole-3-acetic acid (IAA) biosynthesis, trehalose production, siderophore production, acetoin synthesis, and phosphate solubilization were determined. Moreover, genes that contribute to the environmental fitness of UW4 were also observed including genes responsible for heavy metal resistance such as nickel, copper, cadmium, zinc, molybdate, cobalt, arsenate, and chromate. Whole-genome comparison with other completely sequenced Pseudomonas strains and phylogeny of four concatenated “housekeeping” genes (16S rRNA, gyrB, rpoB and rpoD) of 128 Pseudomonas strains revealed that UW4 belongs to the fluorescens group, jessenii subgroup.     

Biotization of Arabidopsis thaliana with Pseudomonas putida and assessment of its positive effect on in vitro growth

Determination of the optimal inoculation method and concentration to use for plant-bacteria interaction studies is important in many cases, such as the phytoremediation of heavy metals and other toxic compounds in contaminated areas. The aim of this study was to compare different concentrations and times of inoculation of Pseudomonas putida with various growth stages of Arabidopsis thaliana in 14-d in vitro cultures. A significant beneficial impact of the bacterium was detected in the shoot length and root weight of seedlings. The highest shoot length and root fresh and dry weights were detected in 14-d and 2 × 10³ cfu mL⁻¹ inoculated samples. In addition, the increase in root weight could be visualized with crystal violet staining, as relatively more root hair and lateral root formation occurred in seedlings inoculated with moderate concentrations of bacteria, possibly due to the ability of P. putida to produce indole-acetic acid. Moreover, the highest photosynthetic pigment accumulation was obtained with the highest bacterial inoculum (2 × 10⁶ cfu mL⁻¹), which was tested in 0- or 3-d-old seedlings. Rhizospheric bacterial colonization was also visualized with GFP-labeled bacteria by confocal microscopy. These results showed that biotization of A. thaliana with P. putida KT2440 did not cause severe oxidative stress in seedlings, because H₂O₂ accumulation levels together with CAT and POX activities were not significantly induced. Therefore, this strain could be used for several applications based on plant-bacteria interactions.

     

Effect of Wild-Type and Mutant Plant Growth-Promoting Rhizobacteria on the Rooting of Mung Bean Cuttings

Mung bean cuttings were dipped in solutions of wild type and mutant forms of the plant growth-promoting rhizobacterium Pseudomonas putida GR12-2 and then incubated for several days until roots formed. The bacteria P. putida GR12-2 and P. putida GR12-2/aux1 mutant do not produce detectable levels of the enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase, whereas P. putida GR12-2/acd36 is an ACC deaminase minus mutant. All bacteria produce the phytohormone indole-3-acetic acid (IAA), and P. putida GR12-2/aux1 overproduces it. Treatment of cuttings with the above-mentioned bacteria affected the rates of ethylene production in the cuttings in a way that can be explained by the combined effects of the activity of ACC deaminase localized in the bacteria and bacterial produced IAA. P. putida GR12-2 and P. putida GR12-2/acd36-treated cuttings had a significantly higher number of roots compared with cuttings rooted in water. In addition, the wild type influenced the development of longer roots. P. putida GR12-2/aux1 stimulated the highest rates of ethylene production but did not influence the number of roots. These results are consistent with the notion that ethylene is involved in the initiation and elongation of adventitious roots in mung bean cuttings. Received October 21, 1998; accepted January 3, 1999     

Inoculation with plant growth-promoting bacteria (PGPB) improves salt tolerance of maize seedling

Our objective was to evaluate the role of plant growth-promoting bacteria to protect maize (Zea mays L.) plants against salt damage. Bacillus aquimaris DY-3 based on their 16S rDNA sequences, the most tolerant to salinity and the synthesis of indole acetic acid was selected for further studies. Strain was inoculated on maize roots growing in sterilized sand under salt stress conditions (1% NaCl). After one week, plant growth was promoted by bacterial inoculation regardless of salt stress and non-salt stress. Chlorophyll content, leaf relative water content, accumulation of proline, soluble sugar and total phenolic compound, and activities of superoxide dismutase, catalase, peroxidase and ascorbate peroxidase were enhanced, while lipid peroxidation levels and Na⁺ content were decreased. The results showed that B. aquimaris DY-3 alleviated the salt stress in maize, likely through the integration of the antioxidant enzymes and the non-antioxidant systems that improve the plant response. Hence, the application of indole acetic acid synthesizing plant growth-promoting bacteria may represent an important alternative approach to decrease the impact of salt stress on crops.     

Comparative effectiveness of <Emphasis Type="Italic">Pseudomonas</Emphasis> and <Emphasis Type="Italic">Serratia</Emphasis> sp. containing ACC-deaminase for improving growth and yield of wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) under salt-stressed conditions

Ethylene synthesis is accelerated in response to various environmental stresses like salinity. Ten rhizobacterial strains isolated from wheat rhizosphere taken from different salt affected areas were screened for growth promotion of wheat under axenic conditions at 1, 5, 10 and 15 dS m⁻¹. Three strains, i.e., Pseudomonas putida (N21), Pseudomonas aeruginosa (N39) and Serratia proteamaculans (M35) showing promising performance under axenic conditions were selected for a pot trial at 1.63 (original), 5, 10 and 15 dS m⁻¹. Results showed that inoculation was effective even in the presence of higher salinity levels. P. putida was the most efficient strain compared to the other strains and significantly increased the plant height, root length, grain yield, 100-grain weight and straw yield up to 52, 60, 76, 19 and 67%, respectively, over uninoculated control at 15 dS m⁻¹. Similarly, chlorophyll content and K⁺/Na⁺ of leaves also increased by P. putida over control. It is highly likely that under salinity stress, 1-aminocyclopropane-1-carboxylic acid-deaminase activity of these microbial strains might have caused reduction in the synthesis of stress (salt)-induced inhibitory levels of ethylene. The results suggested that these strains could be employed for salinity tolerance in wheat; however, P. putida may have better prospects in stress alleviation/reduction.     

The impact of ColRS two-component system and TtgABC efflux pump on phenol tolerance of <Emphasis Type="Italic">Pseudomonas putida</Emphasis> becomes evident only in growing bacteria

Background  

We have recently found that Pseudomonas putida deficient in ColRS two-component system is sensitive to phenol and displays a serious defect on solid glucose medium where subpopulation of bacteria lyses. The latter phenotype is significantly enhanced by the presence of phenol in growth medium. Here, we focused on identification of factors affecting phenol tolerance of the colR-deficient P. putida.     

A Plant Growth-Promoting Bacterium That Decreases Nickel Toxicity in Seedlings

A plant growth-promoting bacterium, Kluyvera ascorbata SUD165, that contained high levels of heavy metals was isolated from soil collected near Sudbury, Ontario, Canada. The bacterium was resistant to the toxic effects of Ni²⁺, Pb²⁺, Zn²⁺, and CrO₄⁻, produced a siderophore(s), and displayed 1-aminocyclopropane-1-carboxylic acid deaminase activity. Canola seeds inoculated with this bacterium and then grown under gnotobiotic conditions in the presence of high concentrations of nickel chloride were partially protected against nickel toxicity. In addition, protection by the bacterium against nickel toxicity was evident in pot experiments with canola and tomato seeds. The presence of K. ascorbata SUD165 had no measurable influence on the amount of nickel accumulated per milligram (dry weight) of either roots or shoots of canola plants. Therefore, the bacterial plant growth-promoting effect in the presence of nickel was probably not attributable to the reduction of nickel uptake by seedlings. Rather, it may reflect the ability of the bacterium to lower the level of stress ethylene induced by the nickel.     

The elicitation of a systemic resistance by Pseudomonas putida BTP1 in tomato involves the stimulation of two lipoxygenase isoforms

Background  

Some non-pathogenic rhizobacteria called Plant Growth Promoting Rhizobacteria (PGPR) possess the capacity to induce in plant defense mechanisms effective against pathogens. Precedent studies showed the ability of Pseudomonas putida BTP1 to induce PGPR-mediated resistance, termed ISR (Induced Systemic Resistance), in different plant species. Despite extensive works, molecular defense mechanisms involved in ISR are less well understood that in the case of pathogen induced systemic acquired resistance.     

Isolation and Characterization of Mutants of the Plant Growth-Promoting Rhizobacterium Pseudomonas putida GR12-2 That Overproduce Indoleacetic Acid

Following transposon Tn5 mutagenesis of the plant growth-promoting rhizobacterium Pseudomonas putida GR12-2, mutants that were able to grow in the presence of the tryptophan analog 5-fluorotryptophan were selected. Seven of the 50 5-fluorotryptophan-resistant mutants overproduced the phytohormone indoleacetic acid (IAA). Of these seven mutants, the highest level of IAA was observed with strain P. putida GR12-2/aux1, which produced four times the amount of indoleacetic acid synthesized by the wild-type strain. Strain P. putida GR12-2/aux1, in contrast to the wild type, lost the ability to stimulate the elongation of the roots of canola seedlings under gnotobiotic conditions. The growth rate, siderophore production, and 1-aminocyclopropane-1-carboxylate deaminase activity of mutant strain P. putida GR12-2/aux1 were identical to those of the wild-type strain. The role of IAA in the mechanism of plant growth stimulation by P. putida GR12-2 and other plant growth-promoting rhizobacteria is discussed.     

Isolation,identification and characterization of soil microbes which degrade phenolic allelochemicals

Aims: To isolate and characterize microbes in the soils containing high contents of phenolics and to dissolve the allelopathic inhibition of plants through microbial degradation. Methods and Results: Four microbes were isolated from plant soils using a screening medium containing p‐coumaric acid as sole carbon source. The isolates were identified by biochemical analysis and sequences of their 16S or 18S rDNA, and designated as Pseudomonas putida 4CD1 from rice (Oryza sativa) soil, Ps. putida 4CD3 from pine (Pinus massoniana) soil, Pseudomonas nitroreducens 4CD2 and Rhodotorula glutinis 4CD4 from bamboo (Bambusa chungii) soil. All isolates degraded 1 g l^?1 of p‐coumaric acid by 70–93% in inorganic and by 99% in Luria‐Bertani solutions within 48 h. They also effectively degraded ferulic acid, p‐hydroxybenzoic acid and p‐hydroxybenzaldehyde. The microbes can degrade p‐coumaric acid and reverse its inhibition on seed germination and seedling growth in culture solutions and soils. Low pHs inhibited the growth and phenolic degradation of the three bacteria. High temperature inhibited the R. glutinis. Co²⁺ completely inhibited the three bacteria, but not the R. glutinis. Cu²⁺, Al³⁺, Zn²⁺, Fe³⁺, Mn²⁺, Mg²⁺ and Ca²⁺ had varying degrees of inhibition for each of the bacteria. Conclusions: Phenolics in plant culture solutions and soils can be decomposed through application of soil microbes in laboratory or controlled conditions. However, modification of growth conditions is more important for acidic and ions‐contaminated media. Significance and Impact of the Study: The four microbes were first isolated and characterized from the soils of bamboo, rice or pine. This study provides some evidence and methods for microbial control of phenolic allelochemicals.     

Growth Enhancement of Chickpea in Saline Soils Using Plant Growth-Promoting Rhizobacteria

Bacterial isolates with the ability to tolerate salinity and plant growth-promoting features were isolated from the saline areas of Gujarat, India, that is, Bhavnagar and Khambat. A total of 176 strains of rhizobacteria were isolated out of which 62 bacterial strains were able to tolerate 1 M NaCl. These were then further studied for their potential plant growth-promoting rhizobacteria characteristics like phosphate solubilization, siderophore production, and IAA production. Twenty-eight isolates of the 62 strains showed good tricalcium phosphate solubilization in solid medium in the range of 9–22 mm and 15 isolates showed good phosphate solubilization in liquid medium in the range of 9–45 μg/ml. Siderophore production was checked in all 15 isolates, and 13 were screened out that produced the hydroxamate type of siderophore in the range of 11–50 mM. Among the 13 isolates, 10 were able to produce indole acetic acid in the range of 10–26 μg/ml after 72 h of incubation. Pot trials were carried out on chickpea under 300 mM NaCl stress using the best five isolates. Plants inoculated with MSC1 or MSC4 isolates showed an increase in the parameters that evaluate plant growth when compared to uninoculated controls. Strains MSC1 and MSC4 were identified as Pseudomonas putida and Pseudomonas pseudoalcaligens, respectively, according to sequence analysis of the 16S rRNA gene.