Enhancing the biological nitrogen fixation of leguminous crops grown under stressed environments

Legumes have the ability to establish a symbiotic interaction with soil bacteria, collectively termed as rhizobia. These bacteria can enhance growth and development of associated crops by transferring atmospheric nitrogen into a form that is available for plant growth or by improving nutrient uptake through modulation of hormone-linked phenomena in inoculated plants. Selection of the effective Rhizobium strain is the most critical aspect used to achieve maximum benefits from this technology. This review aims to focus on recent findings, thereby highlighting the enhancement of plant growth and nitrogen uptake during the symbiosis between rhizobia and leguminous plants under severe conditions. The potential uses of such microorganisms due to their multifaceted beneficial activities are likely to play an important role in modern high intensive agricultural practices.Keywords: Rhizobium spp., legumes, growth promotion, plant protection, sustainable agricultur

Biochemical analysis of induced resistance in chickpea against broomrape (Orobanche foetida) by rhizobia inoculation

This study examined the capacity of Rhizobium sp. strain PchAZM to reduce parasitism of chickpea by Orobanche foetida under greenhouse conditions, and assessed the relative impact of rhizobia on the expression of chickpea defense response against broomrape. Growth chamber experiments using Petri dishes revealed that rhizobia infection on chickpea roots reduced broomrape seed germination, and restricted the broomrape attachment to host roots while retarding tubercle formation and development by the parasite. In pot experiments, chickpea roots inoculated with rhizobia reduced the total number of broomrape by up to 90%. Broomrape necrosis was observed both before and after parasite attachment to inoculated chickpea roots in Petri dishes and pot experiments. Reduction in infection was accompanied by enhanced levels of the defence-related enzymes phenylalanine ammonia lyase (PAL) and peroxidase (POX). Increased levels of phenolics were recorded in the roots of rhizobia-inoculated plants grown in the presence of broomrape. The results suggest that rhizobia could be used for protection of chickpea against O. foetida

Potential of Rhizobia in Improving Nitrogen Fixation and Yields of Legumes

Strong demand for food requires specific efforts by researchers involved in the agricultural sector to develop means for sufficient production. While, agriculture today faces challenges such as soil fertility loss, climate change and increased attacks of pathogens and pests. The production of sufficient quantities in a sustainable and healthy farming system is based on environmentally friendly approaches such as the use of biofertilizers, biopesticides and the return of crop residues. The multiplicity of beneficial effects of soil microorganisms, particularly plant growth promotion (PGP), highlights the need to further strengthen the research and its use in modern agriculture. Rhizobia are considered as PGP comes in symbiosis with legumes taking advantage of nutrients from plant root exudates. When interacting with legumes, rhizobia help in increased plant growth through enriching nutrients by nitrogen fixation, solubilizing phosphates and producing phytohormones, and rhizobia can increase plants’ protection by influencing the production of metabolites, improve plant defense by triggering systemic resistance induced against pests and pathogens. In addition, rhizobia contain useful variations to tolerate abiotic stresses such as extreme temperatures, pH, salinity and drought. The search for rhizobium tolerant strains is expected to improve plant growth and yield, even under a combination of constraints. This chapter summarizes the use of rhizobia in agriculture and its benefits

Dissemination and genetic support of broad-spectrum beta-lactam-resistant Escherichia coli strain isolated from two Tunisian hospitals during 2004-2012.

Background: The dissemination of extended-spectrum \u3b2-lactamase (ESBL)-producing bacteria presented a great concern worldwide. Gram-negative organisms such as Escherichia coli and Klebsiella pneumoniae are the most frequently isolated pathogens responsible for nosocomial infections. Objectives: The aim of this study was to investigate and to follow the emergence of resistance and the characterization of Extended-Spectrum Beta-Lactamases (ESBL) among broad-spectrum beta-lactam-Escherichia coli clinical isolates recovered from the military hospital and Habib Thameur hospital in Tunisia. Methods: A total of 113 E.coli isolates obtained during the period 2004 through 2012 showed a significant degree of multi-resistance. Among these strains, the double-disk synergy test confirmed the ESBL phenotype in 46 isolates. These included 32(70%) strains from Hospital A and 14(30%) from Hospital B. Results: The ESBL was identified as CTX-M-15. The ESBL resistance was transferred by a 60 kb plasmid CTXM-15-producing isolates were unrelated according to the PFGE analysis and characterization of the regions surrounding the blaCTX-M-15 showed the ISEcp1 elements located in the upstream region of the bla gene and 20 of them truncated by IS26. Conclusion: ESBL producing E. coli strains are a serious threat in the community in Tunisia and we should take into consideration any possible spread of such epidemiological resistance

Study of a Natural Mutant SHV-Type -Lactamase, SHV-104, from Klebsiella pneumoniae

Klebsiella pneumoniae ML2011, a multiresistant isolate, was isolated from the Military Hospital of Tunis (Tunisia). The determination of the minimal inhibitory concentrations exhibited by K. pneumoniae ML2011 was performed by Etest. The crude extract of the isolates contains four different -lactamases with pI 5.5, 7.3, 7.6, and 8.6. Only the -lactamases with pI 7.3 and pI 8.6 were transferred by transformation and conjugation experiment. Molecular characterization of these genes was performed by PCR and sequencing. The chromosomal -lactamases are TEM (pI 5.5) and SHV-1 (7.6). CTX-M-28 (pI 8.6) and the novel variant of SHV named SHV-104 (pI 7.3) were encoded by bla gene located on a 50 kb highly conjugative plasmid. The SHV-104 -lactamase was produced in E. coli and purified. Its profile of activity was determined. Compared to SHV-1, SHV-104 contains one mutation, R202S. Their kinetic parameters were similar except for cefotaxime. The analysis of the predicted structure of SHV-104 indicated that the R202S mutation suppresses a salt bridge present in SHV-1. Therefore, the overall flexibility of the protein increased and might improve the hydrolysis of cefotaxime. We can conclude that the multiresistant phenotype of K. pneumoniae ML2011 strain is mainly linked to the production of CTX-M-28 since SHV-104 possesses a narrow spectrum of activity

Purification and Characterization of a New Thermostable, Haloalkaline, Solvent Stable, and Detergent Compatible Serine Protease from Geobacillus toebii Strain LBT 77

A new thermostable, haloalkaline, solvent stable SDS-induced serine protease was purified and characterized from a thermophilic Geobacillus toebii LBT 77 newly isolated from a Tunisian hot spring. This study reveals the potential of the protease from Geobacillus toebii LBT 77 as an additive to detergent with spectacular proprieties described for the first time. The protease was purified to homogeneity by ammonium sulfate precipitation followed by Sephadex G-75 and DEAE-Cellulose chromatography. It was a monomeric enzyme with molecular weight of 30 kDa. The optimum pH, temperature, and NaCl for maximum protease activity were 13.0, 95°C, and 30%, respectively. Activity was stimulated by Ca2+, Mg2+, DTNB, β-mercaptoethanol, and SDS. The protease was extremely stable even at pH 13.25, 90°C, and 30% NaCl and in the presence of hydrophilic, hydrophobic solvents at high concentrations. The high compatibility with ionic, nonionic, and commercial detergents confirms the utility as an additive to cleaning products. Kinetic and thermodynamic characterization of protease revealed Km=1 mg mL−1,  Vmax=217.5 U mL−1, Kcat/Km=99 mg mL−1 S−1, Ea=51.5 kJ mol−1, and ΔG⁎=56.5 kJ mol−1

Screening and characterization of thermo-active enzymes of biotechnological interest produced by thermophilic Bacillus isolated from hot springs in Tunisia

As part of the contribution to the global efforts in research of thermostable enzymes being of industrial interest, we focus on the isolation of thermophilic bacteria from Tunisian hot springs. Among the collection of 161 strains of thermophilic Bacillus isolated from different samples of thermal water in Tunisia, 20% are capable of growing at 100°C and the rest grow at 70°C or above. Preliminary activity tests on media supplemented with enzyme-substrates confirmed that 35 strains produced amylases, 37 - proteases, 43 - cellulases, 31 - xylanases and 37 - mannanases. The study of the effect of temperature on enzyme activity led to determination of the optimal temperatures of activities that vary between 60 and 100°C. Several enzymes were active at high temperatures (80, 90 and 100°C) and kept their activity even at 110°C. Several isolated strains producing enzymes with high optimal temperatures of activity were described for the first time in this study. Both strains B62 and B120 are producers of amylase, protease, cellulase, xylanase, and mannanase. The sequencing of 16S DNA identified isolated strains as Geobacillus kaustophillus, Aeribacillus pallidus, Geobacillus galactosidasus and Geobacillus toebii

Screening and characterization of thermo-active enzymes of biotechnological interest produced by thermophilic Bacillus isolated from hot springs in Tunisia

As part of the contribution to the global efforts in research of thermostable enzymes being of industrial interest, we focus on the isolation of thermophilic bacteria from Tunisian hot springs. Among the collection of 161 strains of thermophilic Bacillus isolated from different samples of thermal water in Tunisia, 20% are capable of growing at 100°C and the rest grow at 70°C or above. Preliminary activity tests on media supplemented with enzyme-substrates confirmed that 35 strains produced amylases, 37 - proteases, 43 - cellulases, 31 - xylanases and 37 - mannanases. The study of the effect of temperature on enzyme activity led to determination of the optimal temperatures of activities that vary between 60 and 100°C. Several enzymes were active at high temperatures (80, 90 and 100°C) and kept their activity even at 110°C. Several isolated strains producing enzymes with high optimal temperatures of activity were described for the first time in this study. Both strains B62 and B120 are producers of amylase, protease, cellulase, xylanase, and mannanase. The sequencing of 16S DNA identified isolated strains as Geobacillus kaustophillus, Aeribacillus pallidus, Geobacillus galactosidasus and Geobacillus toebii