Plant growth promoting bacteria from Crocus sativus rhizosphere

Present study deals with the isolation of rhizobacteria and selection of plant growth promoting bacteria from Crocus sativus (Saffron) rhizosphere during its flowering period (October–November). Bacterial load was compared between rhizosphere and bulk soil by counting CFU/gm of roots and soil respectively, and was found to be ~40 times more in rhizosphere. In total 100 bacterial isolates were selected randomly from rhizosphere and bulk soil (50 each) and screened for in-vitro and in vivo plant growth promoting properties. The randomly isolated bacteria were identified by microscopy, biochemical tests and sequence homology of V1–V3 region of 16S rRNA gene. Polyphasic identification categorized Saffron rhizobacteria and bulk soil bacteria into sixteen different bacterial species with Bacillus aryabhattai (WRF5-rhizosphere; WBF3, WBF4A and WBF4B-bulk soil) common to both rhizosphere as well as bulk soil. Pseudomonas sp. in rhizosphere and Bacillus and Brevibacterium sp. in the bulk soil were the predominant genera respectively. The isolated rhizobacteria were screened for plant growth promotion activity like phosphate solubilization, siderophore and indole acetic acid production. 50 % produced siderophore and 33 % were able to solubilize phosphate whereas all the rhizobacterial isolates produced indole acetic acid. The six potential PGPR showing in vitro activities were used in pot trial to check their efficacy in vivo. These bacteria consortia demonstrated in vivo PGP activity and can be used as PGPR in Saffron as biofertilizers.This is the first report on the isolation of rhizobacteria from the Saffron rhizosphere, screening for plant growth promoting bacteria and their effect on the growth of Saffron plant.     

Diversity of cultivable rhizobacteria across tomato growing regions of Karnataka

Seven hundred and fifty-two rhizobacteria were isolated from 186 rhizosphere soil samples collected across tomato growing regions of Karnataka. Among them, 26% strains were Gram positive and other 74% were Gram negative and dominant being Bacillus and Pseudomonas. Sampling of different locations showed variation in species richness and diversity indices. Similarity matrix computed with Jaccard’s coefficient and principle coordinate analysis to correlate bacterial diversity revealed that rhizobacterial genera of Mysore, Mandya and Kolar soil samples were very closely related and rarefaction curve analysis indicated that these soil samples also harbored higher number of rhizobacteria which included all the genera studied. PGPR trait analysis revealed that most of the rhizobacteria were endowed with more than one beneficial trait which may act individually or simultaneously, and indole acetic acid production and phosphate solubilization are the two predominant traits exhibited by these rhizobacteria. Rhizobacterial isolates also showed a varied level of plant growth promotion traits and offered protection against fungal origin foliar and root pathogens. Among the nine regions studied, Mysore, Mandya and Kolar regions recorded higher percentage of promising PGPRs in comparison with other regions studied of Karnataka.     

Characterization of Rhizobacteria Associated with Weed Seedlings

Rhizobacteria were isolated from seedlings of seven economically important weeds and characterized for potential phytopathogenicity, effects on seedling growth, and antibiosis to assess the possibility of developing deleterious rhizobacteria as biological control agents. The abundance and composition of rhizobacteria varied among the different weed species. For example, fluorescent pseudomonads represented from 11 to 42% of the total rhizobacterial populations from jimsonweed and lambsquarters, respectively. Other bacteria frequently isolated were nonfluorescent pseudomonads, Erwinia herbicola, Alcaligenes spp., and Flavobacterium spp. Only 18% of all isolates were potentially phytopathogenic, based on an Escherichia coli indicator bioassay. However, the proportion of isolates that inhibited growth in seedling assays ranged from 35 to 65% depending on the weed host. Antibiosis was most prevalent among isolates of fluorescent Pseudomonas spp., the activity of which was due to siderophore production in over 75% of these isolates. Overall, rhizobacterial isolates exhibited a complex array of properties that were inconsistent with accepted definitions for plant growth-promoting and deleterious rhizobacteria. It is suggested that for development of effective biological control agents for weed control, deleterious rhizobacteria must be screened directly on host seedlings and must possess several properties including high colonizing ability, specific phytotoxin production, and resistance or tolerance to antibiotics produced by other rhizosphere microorganisms, and they must either synthesize or utilize other bacterial siderophores.     

Isolation and diversity of culturable rhizobacteria associated with economically important crops and uncultivated plants in Québec,Canada

Plants are chronically associated with microorganisms, residing all tissues. Holonomic analysis of diversity of established rhizobacteria in uncultivated plants is scarce. Thus, the present study was conducted to access the root-associated bacterial diversity of 6 crops (maize, canola, soybean, reed canarygrass, alfafa, and miscanthus) and 20 uncultivated plant species in the region of Sainte-Anne-de-Bellevue, Québec, Canada, using pure-culture methods. Based on 16S rRNA gene sequence analysis, 446 bacterial isolates were distributed onto four phyla (Proteobacteria, Firmicutes, Actinobacteria and Bacteroidetes), 32 families and 90 genera. Proteobacteria constituted the largest group of isolates (240), 40% of ectophytic and 61% of endophytic bacteria. Representatives of the genera Bacillus and Pseudomonas dominated in rhizosphere soil; Microbacterium and Pseudomonas were the predominant endophytes. Some genera were associated with specific plant species, such as Stenotrophomonas, Yersinia, Labrys and Luteibacter. Several endophytes were occasionally observed in the rhizosphere, and vice versa. This is the first survey of culturable endophytic bacteria associated with uncultivated plants in Québec. The culturable bacterial community studied herein are assumed to represent a portion of the entire phytomicrobiome of the evaluated plants. Results confirmed that the crops and uncultivated plants of Québec represent an extremely rich reservoir of diverse rhizobacteria.     

Effects of organochlorines on microbial diversity and community structure in Phragmites australis rhizosphere

This study investigated the impacts of an organochlorine (OC, γ-hexachlorocyclohexane and chlorobenzenes) mixture on microbial communities associated to Phragmites australis rhizosphere. Seventy-eight distinct colony morphotypes were isolated, cultivated and analysed by 16S rDNA sequence analysis. Toxicity tests confirmed sensitivity (e.g. Hevizibacter, Acidovorax) or tolerance (e.g. Bacillus, Aeromonas, Pseudomonas, Sphingomonas) of isolates. Rhizosphere analysis by pyrosequencing showed the microbial adaptation induced by OC exposure. Among the most abundant molecular operational taxonomic units, 80 % appeared to be tolerant (55 % opportunist, 25 % unaffected) and 20 % sensitive. P. australis rhizosphere exposed to OCs was dominated by phylotypes related to α-, β- and γ-Proteobacteria. Specific genera were identified which were previously described as chlorinated organic pollutant degraders: Sphingomonas sp., Pseudomonas sp., Devosia sp. and Sphingobium sp. P. australis could be suitable plants to maintain their rhizosphere active microbial population which can tolerate OCs and potentially improve the OC remediation process in part by biodegradation.     

The Effect of Biocontrol Bacteria on Rhizosphere Bacterial Communities Analyzed by Plating and PCR-DGGE

Bacillus subtilis B579, which was isolated from rhizosphere of cucumber, exhibited an excellent biocontrol activity on soil-born pathogens under greenhouse conditions. It could colonize in rhizosphere of cucumber with large number of populations after inoculated in plant growth season. To reveal the effect of high level colonization of B. subtilis B579 on rhizobacteria community structure, cultivation-based analysis coupled with denaturing gradient gel electrophoresis (DGGE) analysis were used to profile the changes of rhizobacteria community structure sampling at 1 week interval. Cultivation-based and DGGE fingerprinting analysis showed significant plant-dependent and seasonal shifts in rhizobacteria populations. Only minimal and transient effects were observed at 4–9 weeks after sowing in samples of B579 treatment, without the pathogen inoculation and showed the best plant growth potential. Sequencing of dominant bands excised from the gel revealed that Streptomyces sp. was the dominate species in soils before and after sowing. Burkholderia sp. was the dominate species in bulk soil, while Bacillus sp. was dominated in rhizosphere within the growth season. Arthrobacter ramosus and Nocardioides sp. were identified as the specific species in samples treated by B579 at the maturity and flowering stages of cucumber.     

Isolation and characterization of KDML105 aromatic rice rhizobacteria producing indole-3-acetic acid: impact of organic and conventional paddy rice practices

Indole-3-acetic acid (IAA) synthesis is a major property of rhizosphere bacteria. The IAA-producing ability of rhizobacteria may be influenced by agricultural management. We therefore evaluated the IAA-producing potential of rhizobacteria isolated during organic rice farming (ORF) and conventional rice farming (CRF) in Thung Kula Rong Hai areas of Thailand. The results indicated that ORF gave a significantly higher percentage of IAA producers (95·8%) than CRF (69·9%). The average IAA values of the ORF isolates were around two times higher than those of the CRF isolates both in the absence (12·8 and 5·8 μg IAA ml⁻¹, respectively) and presence of L-tryptophan (L-Trp) (35·2 and 17·2 μg IAA ml⁻¹, respectively). The 16S rRNA gene sequence analysis indicated that the 23 selected isolates belonged to 8 different genera—Sinomonas sp., Micrococcus sp., Microbacterium sp., Fictibacillus sp., Bacillus sp., Burkholderia sp., Leclercia sp. and Enterobacter sp. Interestingly, only three ORF isolates, i.e. ORF15-20 (Micrococcus sp.), ORF15-21 (Sinomonas sp.) and ORF15-23 (Sinomonas sp.), exhibited high IAA production ability without L-Trp (128·5, 160·8 and 174·7 μg IAA ml⁻¹, respectively). Meanwhile, a slight decrease in IAA production with L-Trp was noticed, suggesting that the L-Trp was not used for the IAA synthesis of these isolates. Biopriming with rhizobacterial isolates significantly enhanced the rate of germination of KDML 105 rice seeds compared to the control.     

Characterization of bacterial communities associated with Brassica napus L. growing on a Zn-contaminated soil and their effects on root growth

The interaction between plant growth-promoting bacteria (PGPB) and plants can enhance biomass production and metal tolerance of the host plants. This work aimed at isolating and characterizing the cultivable bacterial community associated with Brassica napus growing on a Zn-contaminated site, for selecting cultivable PGPB that might enhance biomass production and metal tolerance of energy crops. The effects of some of these bacterial strains on root growth of B. napus exposed to increasing Zn and Cd concentrations were assessed. A total of 426 morphologically different bacterial strains were isolated from the soil, the rhizosphere, and the roots and stems of B. napus. The diversity of the isolated bacterial populations was similar in rhizosphere and roots, but lower in soil and stem compartments. Burkoholderia, Alcaligenes, Agrococcus, Polaromonas, Stenotrophomonas, Serratia, Microbacterium, and Caulobacter were found as root endophytes exclusively. The inoculation of seeds with Pseudomonas sp. strains 228 and 256, and Serratia sp. strain 246 facilitated the root development of B. napus at 1,000 µM Zn. Arthrobacter sp. strain 222, Serratia sp. strain 246, and Pseudomonas sp. 228 and 262 increased the root length at 300 µM Cd.     

Co-inoculation with <Emphasis Type="Italic">Enterobacter</Emphasis> and Rhizobacteria on Yield and Nutrient Uptake by Wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) in the Alluvial Soil Under Indo-Gangetic Plain of India

The aim of this work was to evaluate the effects of co-inoculation with phosphate-solubilizing and nitrogen-fixing rhizobacteria on growth promotion, yield, and nutrient uptake by wheat. Out of twenty-five bacteria isolated from the rhizosphere soils of cereal, vegetable, and agro-forestry plants in eastern Uttar Pradesh, three superior most plant growth-promoting (PGP) isolates were characterized as Serratia marcescens, Microbacterium arborescens, and Enterobacter sp. based on their biochemical and 16S rDNA gene sequencing data and selected them for evaluating their PGP effects on growth and yield of wheat. Among them, Enterobacter sp. and M. arborescens fixed significantly higher amounts (9.32?±?0.57 and 8.89?±?0.58 mg Ng^?1 carbon oxidized, respectively) of atmospheric nitrogen and produced higher amounts (27.06?±?1.70 and 26.82?±?1.63 TP 100 µg mL^?1, respectively) of IAA in vitro compared to S. marcescens (8.32?±?0.39 mg Ng^?1 carbon oxidized and 21.29?±?0.99 TP 100 µg mL^?1). Although both M. arborescens and S. marcescens solubilized remarkable amounts of phosphate from tricalcium phosphate likely through production of organic acids, however, Enterobacter sp. was inactive. The effects of these three rhizobacteria were evaluated on wheat in alluvial soils of the Indo-Gangetic Plain by inoculation of plants with bacterial isolates either alone or in combinations in both pot and field conditions for two successive years. Rhizobacterial inoculation either alone or in consortium of varying combinations significantly (P?≤?0.05) increased growth and yield of wheat compared to mock inoculated controls. A consortium of two or three rhizobacterial isolates also significantly increased plant height, straw yield, grain yield, and test weight of wheat in both pot and field trials compared to single application of any of these isolates. Among the rhizobacterial treatment, co-inoculation of three rhizobacteria (Enterobacter, M. arborescens and S. marcescens) performed best in promotion of growth, yield, and nutrient (N, P, Cu, Zn, Mn, and Fe) uptake by wheat. Taken together, our results suggest that co-inoculation of Enterobacter with S. marcescens and M. arborescens could be used for preparation of an effective formulation of PGP consortium for eco-friendly and sustainable production of wheat.     

Plant growth promoting traits of phosphate-solubilizing rhizobacteria isolated from apple trees in trans Himalayan region of Himachal Pradesh

Two hundred and six phosphate-solubilizing rhizobacteria (PSB) were isolated from rhizosphere soil (RS) and root endosphere (ER) of apple trees from different sites of four locations viz., Chamba, Shimla, Kinnaur and Kullu of Himachal Pradesh, Northern India, and were screened for plant growth promoting traits (PGPTs) by using culture dependent procedures. Indole acetic acid (IAA) production was detected in 50 isolates (24.2 %), siderophore synthesis in 53 isolates (25.7 %), hydrocyanic acid (HCN) in 40 isolates (19.4 %) and percentage growth inhibition against Dematophora necatrix in 61 isolates (29.6 %). Overall, 54.3 % of PSB isolates from RS and 64.4 % from ER showed none of the PGPTs tested. Among the PSB showing PGPTs, 10.6 % had single trait and 30.6 % had multiple traits showing two (10.7 %), three (14.1 %) and four (5.8 %) types of PGPTs. The Shannon–Weaver diversity index (H′) revealed that PGPT-possessing PSBs in RS were more abundant than ER. Clustering analysis by principal component analysis showed that ER was most important factor influencing the ecological distribution and physiological characterization of PGPT-possessing PSB. There was a positive correlation (0.94, p < 0.05) between HCN and antifungal activity producers, and IAA and antifungal activity producers (0.99, p < 0.05). Significant positive correlation (0.42, p < 0.05) between HCN producers and altitude was also noted.     

Characterization of a novel antarctic plant growth-promoting bacterial strain and its interaction with antarctic hair grass (Deschampsia antarctica Desv)

Deschampsia antarctica is the only hair grass that has been able to successfully colonize the Antarctic continent. However, there is little research on the role of microorganisms associated with the rhizosphere that may participate in its growth and development. The objective of this research was to characterize a psychrotolerant bacterial strain isolated from the rhizosphere of D. antarctica. Biochemical and molecular studies were performed to characterize this bacterium. It was determined that this strain secretes a neutral polysaccharide that presents different compositions at different temperatures (4 and 20 °C). Based on biochemical and phylogenetic analyses, the Antarctic rhizobacterium could be a new species of Pseudomonas. To determine their ability to solubilize different sources of inorganic phosphate, qualitative and quantitative analyses were conducted to determine P released at 4 °C. The Antarctic strain of Pseudomonas sp. was able to solubilize all sources of phosphates, and 34.2 mg P/L was released from rock phosphate. Growth physiological parameters were evaluated for seedlings of D. antarctica inoculated with the rhizobacteria. It was found that the bacterial inoculation promoted plant root development. SEM analysis of the roots showed that the bacterium is mainly located in the root hairs of D. antarctica.     

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

Background and Aims

This 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.

Methods

299 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.

Results

All 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.

Conclusions

Enterobacter 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.     

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

Background and Aims

Several 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.

Methods

Bacterial 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.

Results

In 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.

Conclusions

The 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.     

Diversity and metabolic potential of culturable bacteria from the rhizosphere of Turkish tea grown in acidic soils

The purpose of this study was to investigate the diversity of cultivable phosphate solubilising (PSB) and total bacteria originated from 384 rhizospheric acidic soils samples of tea plants grown at 32 locations. Over 900 rhizoplane bacteria were randomly selected from agar-solidified trypticase soy broth, and identified using fatty acid methyl ester (FAME) profiles. Based on FAME profiles, 53 bacterial genera were identified with a similarity index >0.3, but 60.3% of the identified isolates belonged to five genera: Bacillus (34.6%), Pseudomonas (8.9%), Stenotrophomonas (6.1%), Paenibacillus (5.9%) and Arthrobacter (4.8%). The bacilli group comprised many different species, with the most abundant being B. cereus, B. megaterium and B. sphaericus. The main identified Pseudomonads included P. fluorescens, P. putida, and P. alcaligenes. About 30.4% of the bacterial isolates could not be classified to genus since their similarity indices were <0.3 indicating no close matches. Most of the total and P-solubilizing bacteria isolated were Gram positive (61.3 and 52.3%), and Gram negative constituted only 38.7 and 47.7%. Out of the 214 PSB from a pool of 506 bacterial isolates recovered on the selective media from the rhizosphere of tea, 74 of them were characterized by carbon sources using BIOLOG^M GN2 and GP2 plates. Bacillus, Pseudomonas, Paenibacillus and Stenotrophomonas genera were the most prominent P-solubilizing groups in the rhizosphere and soil populations analyzed. B. cereus, P. fluorescens, S. maltophilia, B. megaterium, P. putida, B. sphaericus and Paenibacillus polymyxa were the most frequent P-solubilizing species in the acidic tea rhizosohere soils. Selected Gram-positive PSB appeared to favour carbohydrates, and Gram-negative bacteria appeared to favour carboxylic acids, amino acids and carbohydrates as carbon sources. Selected phosphate solubilizing acid tolerant strains showed high variability in utilizing various carbon sources.     

Molecular diversity of 1-aminocyclopropane-1-carboxylate (ACC) deaminase producing PGPR from wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) rhizosphere

Aims

The present study was planned to investigate the diversity of 1-aminocyclopropane-1-carboxylate (ACC) deaminase producing bacteria from the rhizosphere of wheat plants and subsequent evaluation of selected PGPR on growth enhancement of wheat seedlings under drought and saline conditions.

Methods

ACC deaminase producing plant growth promoting rhizobacteria (PGPR) were isolated from the rhizosphere of wheat and identified using 16S rRNA gene sequence analysis. Isolates were evaluated for various direct and indirect plant growth promoting (PGP) traits. Plant inoculation experiment was conducted using isolates IG 19 and IG 22 in wheat to assess their plant growth promotion potential under salinity and drought stress.

Results

Thirty-eight ACC deaminase producing PGPR were isolated which belonged to 12 distinct genera and falling into four phyla γ-proteobacteria, β-proteobacteria, Flavobacteria and Firmicutes. Klebsiella sp. was the most abundant genera and followed by Enterobacter sp. The isolates exhibited ACC deaminase activities ranging from 0.106–0.980 μM α- ketobutyrate μg protein^?1 h^?1. The isolates showed multiple PGP traits such as IAA production, phosphate, zinc, potassium solubilization and siderophore production. Enterobacter cloacae (IG 19) and Citrobacter sp. (IG 22) inoculated wheat seedlings showed notable increases in fresh and dry biomass under non-stress as well as under stressed condition.

Conclusion

To the best of our knowledge this is the first report of presence of ACC deaminase activity and other PGP traits from the genus Citrobacter and Empedobacter. Our finding revealed that the γ-proteobacteria group dominated the wheat rhizosphere. Plant inoculation with PGPR could be a sustainable approach to alleviate abiotic stresses in wheat plants. These native PGPR isolates could be used as potential biofertilizers for sustainable agriculture.

     

Ecophysiological properties of cultivable heterotrophic bacteria and yeasts dominating in phytocenoses of Galindez Island,maritime Antarctica

Antarctic plants are stable specific microenvironments for microbial colonization that are still less explored. In this study, we investigated cultivable heterotrophic bacteria and yeasts dominating in plant samples collected from different terrestrial biotopes near Ukrainian Antarctic Base on Galindez Island, maritime Antarctica. Phylogenetic analysis revealed affiliation of the bacterial isolates to genera Pseudomonas, Stenotrophomonas, Brevundimonas, Sporosarcina, Dermacoccus, Microbacterium, Rothia and Frondihabitans, and the yeast isolates to genera Rhodosporidium, Cryptococcus, Leucosporidiella, Candida and Exophiala. Some ecophysiological properties of isolated strains were determined that are important in response to different stresses such as psychro- and halotolerance, UV-resistance and production of hydrolytic enzymes. The majority of isolates (88 %) was found to be psychrotolerant; all are halotolerant. Significant differences in survival subsequent to UV-C radiation were observed among the isolates, as measured by culturable counts. For the bacterial isolates, lethal doses in the range 80–600 J m^?2 were determined, and for the yeast isolates—in the range 300–1,000 J m^?2. Dermacoccus profundi U9 and Candida davisiana U6 were found as most UV resistant among the bacterial and yeast isolates, respectively. Producers of caseinase, gelatinase, β-glucosidase, and cellulase were detected. To the best of our knowledge, this is the first report on isolation of UV resistant strain D. profundi, and Frondihabitans strain from Antarctica, and on detection of cellulase activity in Antarctic yeast strain C. davisiana. The results obtained contribute to clarifying adaptation strategies of Antarctic microbiota and its possible role in functional stability of Antarctic biocenoses. Stress tolerant strains were detected that are valuable for ecological and applied studies.     

Rhizobacteria of Cotton and Their Repression of Seedling Disease Pathogens

During the 1983 field season, the rhizobacteria (including organisms from rhizosphere soil and the root rhizoplane) of cotton plants at one location in Mississippi were inventoried at different plant growth stages. Isolates (1,000) were identified to the genus level and characterized for repression of Pythium ultimum and Rhizoctonia solani. Cotton seedlings were initially colonized by bacteria of many different genera, and populations quickly reached 10⁸ CFU/g of root tissue. As the season progressed, the bacterial populations declined as root mass increased and the roots became more woodlike in consistency. Fluorescent pseudomonads were the most numerous gram-negative rhizobacterial isolates of those that were randomly collected and identified, and they provided the largest number of isolates with fungal repressive activity. Several other gram-negative bacterial genera were recovered throughout the growing season, and some gram-positive bacteria were also isolated routinely, but at lower numbers. There was no correlation between the proportion of rhizobacterial isolates that possessed fungal repressive activity and the plant growth stage from which the isolates were obtained. Approximately twice as many bacterial isolates demonstrated fungal repression in the agar assay compared with the inplanta assay, and isolates were found more frequently with fungal repressive activity against P. ultimum than against R. solai.     

Culture-Dependent and Independent Studies of Microbial Diversity in Highly Copper-Contaminated Chilean Marine Sediments

Cultivation and molecular-based approaches were used to study microbial diversity in two Chilean marine sediments contaminated with high (835 ppm) and very high concentrations of copper (1,533 ppm). The diversity of cultivable bacteria resistant to copper was studied at oxic and anoxic conditions, focusing on sulfate-, thiosulfate-, and iron-reducing bacteria. For both sediments, the cultivable bacteria isolated at oxic conditions were mostly affiliated to the genus Bacillus, while at anoxic conditions the majority of the cultivable bacteria found were closely related to members of the genera Desulfovibrio, Sphingomonas, and Virgibacillus. Copper resistance was between 100 and 400 ppm, with the exception of a strain affiliated to members of the genus Desulfuromonas, which was resistant up to 1,000 ppm of copper. In parallel, cloning and sequencing of 16S rRNA was performed to study the total bacterial diversity in the sediments. A weak correlation was observed between the isolated strains and the 16S rRNA operational taxonomic units detected. The presence of copper resistance genes (copA, cusA, and pcoA) was tested for all the strains isolated; only copA was detected in a few isolates, suggesting that other copper resistance mechanisms could be used by the bacteria in those highly copper-contaminated sediments.     

Isolation and characterization of aerobic,culturable, arsenic-tolerant bacteria from lead–zinc mine tailing in southern China

Bioremediation of arsenic (As) pollution is an important environmental issue. The present investigation was carried out to isolate As-resistant novel bacteria and characterize their As transformation and tolerance ability. A total of 170 As-resistant bacteria were isolated from As-contaminated soils at the Kangjiawan lead–zinc tailing mine, located in Hunan Province, southern China. Thirteen As-resistant isolates were screened by exposure to 260 mM Na₂HAsO₄·7H₂O, most of which showed a very high level of resistance to As⁵⁺ (MIC?≥?600 mM) and As³⁺ (MIC?≥?10 mM). Sequence analysis of 16S rRNA genes indicated that the 13 isolates tested belong to the phyla Firmicutes, Proteobacteria and Actinobacteria, and these isolates were assigned to eight genera, Bacillus, Williamsia, Citricoccus, Rhodococcus, Arthrobacter, Ochrobactrum, Pseudomonas and Sphingomonas. Genes involved in As resistance were present in 11 of the isolates. All 13 strains transformed As (1 mM); the oxidation and reduction rates were 5–30% and 10–51.2% within 72 h, respectively. The rates of oxidation by Bacillus sp. Tw1 and Pseudomonas spp. Tw224 peaked at 42.48 and 34.94% at 120 h, respectively. For Pseudomonas spp. Tw224 and Bacillus sp. Tw133, the highest reduction rates were 52.01% at 48 h and 48.66% at 144 h, respectively. Our findings will facilitate further research into As metabolism and bioremediation of As pollution by genome sequencing and genes modification.     

Beneficial native bacteria improve survival and mycorrhization of desert truffle mycorrhizal plants in nursery conditions

Sixty-four native bacterial colonies were isolated from mycorrhizal roots of Helianthemum almeriense colonized by Terfezia claveryi, mycorrhizosphere soil, and peridium of T. claveryi to evaluate their effect on mycorrhizal plant production. Based on the phylogenetic analysis of the 16S rDNA partial sequence, 45 different strains from 17 genera were gathered. The largest genera were Pseudomonas (40.8 % of the isolated strains), Bacillus (12.2 % of isolated strains), and Varivorax (8.2 % of isolated strains). All the bacteria were characterized phenotypically and by their plant growth-promoting rhizobacteria (PGPR) traits (auxin and siderophore production, phosphate solubilization, and ACC deaminase activity). Only bacterial combinations with several PGPR traits or Pseudomonas sp. strain 5, which presents three different PGPR traits, had a positive effect on plant survival and growth. Particularly relevant were the bacterial treatments involving auxin release, which significantly increased the root-shoot ratio and mycorrhizal colonization. Moreover, Pseudomonas mandelii strain 29 was able to considerably increase mycorrhizal colonization but not plant growth, and could be considered as mycorrhiza-helper bacteria. Therefore, the mycorrhizal roots, mycorrhizosphere soil, and peridium of desert truffles are environments enriched in bacteria which may be used to increase the survival and mycorrhization in the desert truffle plant production system at a semi-industrial scale.