Asma Imran

Isolation and characterization of plant growth-promoting rhizobacteria from wheat rhizosphere and their effect on plant growth promotion

The present study was conducted to characterize the native plant growth promoting (PGP) bacteria from wheat rhizosphere and root-endosphere in the Himalayan region of Rawalakot, Azad Jammu and Kashmir (AJK), Pakistan. Nine bacterial isolates were purified, screened in vitro for PGP characteristics and evaluated for their beneficial effects on the early growth of wheat (Triticum aestivum L.). Among nine bacterial isolates, seven were able to produce indole-3- acetic acid in tryptophan-supplemented medium; seven were nitrogen fixer, and four were able to solubilize inorganic phosphate in vitro. Four different morphotypes were genotypically identified based on IGS-RFLP fingerprinting and representative of each morphotype was identified by 16S rRNA gene sequencing analysis except Gram-positive putative Bacillus sp. Based on 16S rRNA gene sequence analysis, bacterial isolates AJK-3 and AJK-9 showing multiple PGP-traits were identified as Stenotrophomonas spp. while AJK-7 showed equal homologies to Acetobacter pasteurianus and Stenotrophomonas specie. Plant inoculation studies indicated that these Plant growth-promoting rhizobacteria (PGPR) strains provided a significant increase in shoot and root length, and shoot and root biomass. A significant increase in shoot N contents (up to 76%) and root N contents (up to 32%) was observed over the un-inoculated control. The study indicates the potential of these PGPR for inoculums production or biofertilizers for enhancing growth and nutrient content of wheat and other crops under field conditions. The study is the first report of wheat associated bacterial diversity in the Himalayan region of Rawalakot, AJK.

Cr-resistant rhizo- and endophytic bacteria associated with Prosopis juliflora and their potential as phytoremediation enhancing agents in metal-degraded soils.

Prosopis juliflora is characterized by distinct and profuse growth even in nutritionally poor soil and environmentally stressed conditions and is believed to harbor some novel heavy metal-resistant bacteria in the rhizosphere and endosphere. This study was performed to isolate and characterize Cr-resistant bacteria from the rhizosphere and endosphere of P. juliflora growing on the tannery effluent contaminated soil. A total of 5 and 21 bacterial strains were isolated from the rhizosphere and endosphere, respectively, and were shown to tolerate Cr up to 3000 mg l−1. These isolates also exhibited tolerance to other toxic heavy metals such as, Cd, Cu, Pb, and Zn, and high concentration (174 g l−1) of NaCl. Moreover, most of the isolated bacterial strains showed one or more plant growth-promoting activities. The phylogenetic analysis of the 16S rRNA gene showed that the predominant species included Bacillus, Staphylococcus and Aerococcus. As far as we know, this is the first report analyzing rhizo- and endophytic bacterial communities associated with P. juliflora growing on the tannery effluent contaminated soil. The inoculation of three isolates to ryegrass (Lolium multiflorum L.) improved plant growth and heavy metal removal from the tannery effluent contaminated soil suggesting that these bacteria could enhance the establishment of the plant in contaminated soil and also improve the efficiency of phytoremediation of heavy metal-degraded soils.

Plant–bacteria partnerships for the remediation of persistent organic pollutants

High toxicity, bioaccumulation factor and widespread dispersal of persistent organic pollutants (POPs) cause environmental and human health hazards. The combined use of plants and bacteria is a promising approach for the remediation of soil and water contaminated with POPs. Plants provide residency and nutrients to their associated rhizosphere and endophytic bacteria. In return, the bacteria support plant growth by the degradation and detoxification of POPs. Moreover, they improve plant growth and health due to their innate plant growth-promoting mechanisms. This review provides a critical view of factors that affect absorption and translocation of POPs in plants and the limitations that plant have to deal with during the remediation of POPs. Moreover, the synergistic effects of plant–bacteria interactions in the phytoremediation of organic pollutants with special reference to POPs are discussed.

Phytoremediation: recent advances in plant-endophytic synergistic interactions

BackgroundPlant-endophyte partnership provides an excellent gateway toward restoration of contaminated ecosystems. The interactions between both partners can be manipulated by introducing novel catabolic/metabolic genes into host plant or endophytic bacteria genomes. The potential of the engineered organisms to degrade or accumulate contaminants is much wider than their wild relatives.ScopeThis review covers the recent developments for engineering catabolic/metabolic genes from a wide range of sources into plants or endophytic bacteria for the development of modified plant-endophyte interactions. Genetic alteration of plants promises enhanced catabolism by plant’s own enzymatic machinery or greater contaminant uptake/accumulation for subsequent in planta detoxification by complementary endophytes. On the other hand, bacteria may also be engineered to enhance the potential for degradation or alteration of catabolic pathways, either to protect the host plant against phytotoxicity or to improve the overall efficiency of phytoremediation in planta, a situation especially suitable when hydrophilic compounds fail to be degraded by rhizospheric microbes due to the rapid uptake by plants. This is followed by discussion on kinetic parameters controlling phytoremediation.ConclusionsIt is hypothesized that transgenic approach can result in synergistic and effective plant-endophyte partnerships for wider-range and enhanced capabilities of degrading and/or detoxifying contaminants.

Ochrobactrum ciceri sp. nov., isolated from nodules of Cicer arietinum

A Gram-staining-negative, aerobic, rod-shaped, non-spore-forming bacterial strain, Ca-34(T), was isolated from nodules of chickpea (Cicer arietinum) in Pakistan and studied for its taxonomic affiliation. The almost full-length 16S rRNA gene sequence showed highest similarities to those of strains of the genus Ochrobactrum. Based on results of MALDI-TOF MS and 16S rRNA gene sequence similarity (98.6 %), strain Ca-34(T) and Ochrobactrum intermedium LMG 3301(T) are phylogenetic neighbours; the two strains shared DNA-DNA relatedness of 64 %. The fatty acid profile [predominantly C(18 : 1)omega7c (67.7 %) and C(19 : 0) cyclo omega8c (19.6 %)] also supported the genus affiliation. Metabolically, strain Ca-34(T) differed from other type strains of Ochrobactrum in many reactions and from all type strains in testing positive for gelatin hydrolysis and in testing negative for assimilation of alaninamide and l-threonine. Based on phenotypic and genotypic data, we conclude that strain Ca-34(T) represents a novel species, for which we propose the name Ochrobactrum ciceri sp. nov. (type strain Ca-34(T) =DSM 22292(T) =CCUG 57879(T)).

Genetic, physiological and biochemical characterization of Bacillus sp. strain RMB7 exhibiting plant growth promoting and broad spectrum antifungal activities

BackgroundPlant growth promoting rhizobacteria (PGPR) are functionally diverse group of bacteria having immense potential as biofertilizers and biopesticides. Depending upon their function, they may serve as partial replacements for chemical fertilizer or pesticides as an eco-friendly and cost-effective alternatives as compared to their synthetic counterparts. Therefore, isolation, characterization and practical evaluation of PGPRs having the aforementioned multifaceted beneficial characteristics, are essentially required. This study describes the detailed polyphasic characterization of Bacillus sp. strain RMB7 having profound broad spectrum antifungal activity and plant growth promoting potential.ResultsBased on 16S rRNA gene sequencing, strain RMB7 was identified as Bacillus specie. This strain exhibited the production of 8 mg. L−1of indole-3-acetic acid (IAA) in tryptophan-supplemented medium. It was able to solubilize 50.6 mg. L-1 tri-calcium phosphate, reduced 601ηmol acetylene h-1/vial and inhibited >70% growth of nine fungal phytopathogens tested in vitro. Under natural pathogen pressure, inoculation with strain RMB7 and RMB7-supernatant conferred resistance by arugula plant against Pythium irregulare with a concurrent growth improvement over non-inoculated plants. The T-RFLP analysis based on 16S rRNA gene showed that inoculation with RMB7 or its supernatant have a major impact on the indigenous rhizosphere bacterial population. Mass spectrometric analysis revealed the production of lipopeptide surfactins as well as iturin A presence in crude extract of RMB7. PCR-amplification further confirmed the presence of genes involved in the biosynthesis of these two bioactive lipopeptide compounds.ConclusionsThe data show that Bacillus sp. strain RMB7 has multifaceted beneficial characteristics. It may be an ideal plant growth promoting as well as biocontrol agent, for its integrated use in disease and nutrient management strategies.

Isolation and characterization of a β-propeller gene containing phosphobacterium Bacillus subtilis strain KPS-11 for growth promotion of potato (Solanum tuberosum L.)

Phosphate-solubilizing and phytate-mineralizing bacteria collectively termed as phosphobacteria provide a sustainable approach for managing P-deficiency in agricultural soils by supplying inexpensive phosphate to plants. A phosphobacterium Bacillus subtilis strain KPS-11 (Genbank accession no. KP006655) was isolated from potato (Solanum tuberosum L.) rhizosphere and characterized for potato plant growth promoting potential. The strain utilized both Ca-phosphate and Na-phytate in vitro and produced 6.48 μg mL-1 indole-3-acetic acid in tryptophan supplemented medium. P-solubilization after 240 h was 66.4 μg mL-1 alongwith the production of 19.3 μg mL-1 gluconic acid and 5.3 μg mL-1 malic acid. The extracellular phytase activity was higher (4.3 × 10-10 kat mg-1 protein) than the cell-associated phytase activity (1.6 × 10-10 kat mg-1 protein). B. subtilis strain KPS-11 utilized 40 carbon sources and showed resistance against 20 chemicals in GENIII micro-plate system demonstrating its metabolic potential. Phytase-encoding gene β-propeller (BPP) showed 92% amino acid similarity to BPP from B. subtilis (accession no.WP_014114128.1) and 83% structural similarity to BPP from B. subtilis (accession no 3AMR_A). Potato inoculation with B. subtilis strain KPS-11 increased the root/shoot length and root/shoot weight of potato as compared to non-inoculated control plants. Moreover, rifampicin-resistant derivative of KPS-11 were able to survive in the rhizosphere and on the roots of potato up to 60 days showing its colonization potential. The study indicates that B. subtilis strain KPS-11 can be a potential candidate for development of potato inoculum in P-deficient soils.

Ecology of bacterial endophytes associated with wetland plants growing in textile effluent for pollutant-degradation and plant growth-promotion potentials

In this study, 41 culturable endophytic bacteria were isolated from the roots and shoots of three wetland plants, Typha domingensis, Pistia stratiotes and Eichhornia crassipes, and identified by 16S rRNA gene sequencing. Textile effluent-degrading and plant growth-promoting activities of these endophytes were determined. The analysis of endophytic bacterial communities indicated that plant species had a pronounced effect on endophytic bacterial association and maximum endophytes (56.5%) were associated with T. domingensis. These endophytic bacteria mainly belonged to different species of the genera Bacillus (39%), Microbacterium (12%) and Halomonas (12%). Eight of the 41 strains showing maximum efficiency of textile effluent degradation also exhibited plant growth-promoting activities such as production of indole-3-acetic acid and siderophore, presence of 1-amino-cyclopropane-1-carboxylic acid deaminase, and solubilization of inorganic phosphorous. This is the first study describing the diversity and plant-beneficial characteristics of the textile effluent-degrading endophytic bacteria associated with wetland plants. T. domingensis showed better growth in textile effluent and also hosted maximum number of endophytic bacteria in roots and shoots. The interactions between T. domingensis and its associated endophytic bacteria could be exploited to enhance the efficiency of constructed wetlands during the remediation of industrial effluent.

Differential Response of Potato Toward Inoculation with Taxonomically Diverse Plant Growth Promoting Rhizobacteria

Rhizosphere engineering with beneficial plant growth promoting bacteria offers great promise for sustainable crop yield. Potato is an important food commodity that needs large inputs of nitrogen and phosphorus fertilizers. To overcome high fertilizer demand (especially nitrogen), five bacteria, i.e., Azospirillum sp. TN10, Agrobacterium sp. TN14, Pseudomonas sp. TN36, Enterobacter sp. TN38 and Rhizobium sp. TN42 were isolated from the potato rhizosphere on nitrogen-free malate medium and identified based on their 16S rRNA gene sequences. Three strains, i.e., TN10, TN38, and TN42 showed nitrogen fixation (92.67–134.54 nmol h-1mg-1 protein), while all showed the production of indole-3-acetic acid (IAA), which was significantly increased by the addition of L-tryptophan. Azospirillum sp. TN10 produced the highest amount of IAA, as measured by spectrophotometry (312.14 μg mL-1) and HPLC (18.3 μg mL-1). Inoculation with these bacteria under axenic conditions resulted in differential growth responses of potato. Azospirillum sp. TN10 incited the highest increase in potato fresh and dry weight over control plants, along with increased N contents of shoot and roots. All strains were able to colonize and maintain their population densities in the potato rhizosphere for up to 60 days, with Azospirillum sp. and Rhizobium sp. showing the highest survival. Plant root colonization potential was analyzed by transmission electron microscopy of root sections inoculated with Azospirillum sp. TN10. Of the five test strains, Azospirillum sp. TN10 has the greatest potential to increase the growth and nitrogen uptake of potato. Hence, it is suggested as a good candidate for the production of potato biofertilizer for integrated nutrient management.

Differential response of kabuli and desi chickpea genotypes toward inoculation with PGPR in different soils

Pakistan is among top three chickpea producing countries but the crop is usually grown on marginal lands without irrigation and fertilizer application which significantly hampers its yield. Soil fertility and inoculation with beneficial rhizobacteria play a key role in nodulation and yield of legumes. Four kabuli and six desi chickpea genotypes were, therefore, evaluated for inoculation response with IAA-producing Ochrobactrum ciceri Ca-34T and nitrogen fixing Mesorhizobium ciceri TAL-1148 in single and co-inoculation in two soils. The soil type 1 was previously unplanted marginal soil having low organic matter, P and N contents compared to soil type 2 which was a fertile routinely legume-cultivated soil. The effect of soil fertility status was pronounced and fertile soil on average, produced 31% more nodules, 62% more biomass and 111% grain yield than marginal soil. Inoculation either with O. ciceri alone or its co-inoculation with M. ciceri produced on average higher nodules (42%), biomass (31%), grains yield (64%) and harvest index (72%) in both chickpea genotypes over non-inoculated controls in both soils. Soil 1 showed maximum relative effectiveness of Ca-34T inoculation for kabuli genotypes while soil 2 showed for desi genotypes except B8/02. Desi genotype B8/02 in soil type 1 and Pb-2008 in soil type 2 showed significant yield increase as compared to respective un-inoculated controls. Across bacterial inoculation treatments, grain yield was positively correlated to growth and yield contributing parameters (r = 0.294* to 0.838*** for desi and r = 0.388* to 0.857** for kabuli). PCA and CAT-PCA analyses clearly showed a site-specific response of genotype x bacterial inoculation. Furthermore, the inoculated bacterial strains were able to persist in the rhizosphere showing colonization on root and within nodules. Present study shows that plant growth promoting rhizobacteria (PGPR) inoculation should be integrated with national chickpea breading program in Pakistan especially for marginal soils. Furthermore, the study shows the potential of phytohormone producing strain Ca-34T as promising candidate for development of biofertilizer alongwith nodulating strains to get sustainable yield of kabuli and desi chickpea with minimum inputs at marginal land.