Effect of Potassium Deficiency on Physiological Responses and Anatomical Structure of Basil, Ocimum basilicum L.

The aim of this study was to investigate the effect of a variable supply of potassium to culture medium on physiological and anatomical parameters (histological sections at the third internode) in basil, Ocimum basilicum. Thirty-four-day-old plants grown on basic nutrient medium were divided into four batches and grown on media with varying doses of potassium: 0.375 mM, 0.250 mM, 0.125 mM and 0 mM K+. After 64 days of culture, a final harvest was performed. The results showed that root and shoot growth in basil was decreased with decreased K+ concentration. This restriction was associated with a reduction in root elongation and leaf expansion, which was coupled with a decrease in chlorophyll and carotenoid contents. The estimation of electrolyte leakage reveals that this parameter was increased by potassium deficiency. With respect to total polyphenol and flavonoid contents, only the third leaf-stage extracts exhibited a decrease under low-K+ conditions. However, variability in response of phenolic compounds was recorded depending on the organ and the K+ concentration in the medium. Stem cross sections of potassium-deficient basil plants revealed a decrease in the diameter of these organs, which can be attributed to a restriction of the extent of different tissue territories (cortex and medulla), as well as by a reduction in cell size. These effects were associated with a decrease in the number of conducting vessels and an increase in the number of woody fibers.

Phytochemical Characterization and Bioactivity of Asparagus acutifolius: A Focus on Antioxidant, Cytotoxic, Lipase Inhibitory and Antimicrobial Activities.

The phytochemical composition of leaves, stems, pericarps and rhizomes ethanolic extracts of Asparagus acutifolius were characterized by HPLC-DAD-MS. A. acutifolius samples contain at least eleven simple phenolics, one flavonon, two flavonols and six steroidal saponins. The stem extracts showed the highest total phenolic acid and flavonoid contents, where cafeic acid and rutin were the main compounds. No flavonoids were detected in the leaf, pericarp or rhizome while caffeic acid and ferulic acid were the predominant. Steroidal saponins were detected in the different plant parts of A. acutifolius, and the highest contents were found in the rhizome extracts. The stem extracts exhibited the highest antioxidant activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) and the highest 2,2-azino-bis (3 ethylbenzothiazoline-6-sulphonic acid) (ABTS) scavenging activity was found in the pericarp extracts. The rhizome and leaf extracts showed a potent cytotoxic activity against HCT-116 and HepG2 cell lines. Moreover, the pericarp and rhizome extracts revealed a moderate lipase inhibitory activity. The leaf and rhizome extracts were screened for their antimicrobial activity against human pathogenic isolates. The leaf extract exhibited a powerful inhibitory activity against all the bacteria and fungi tested.

Host plant defenses of black (Solanum nigrum L.) and red nightshade ( Solanum villosum Mill.) against specialist Solanaceae herbivore Leptinotarsa decemlineata (Say).

Black nightshade (Solanum nigrum, S. nigrum L.) and red nightshade ( Solanum villosum, S. villosum Mill.) are medicinal plants from the Solanaceae family that synthesize glycoalkaloids and other secondary metabolites. To recognize the potential insecticide activity of these compounds, leaf extracts (containing glycoalkaloid and methanol fractions) were tested for enzyme inhibition, antifeedant activity and toxicity. For in-vitro glutathione S-transferase (GST) inhibition activity, we used insecticide-resistant Colorado potato beetle, Leptinotarsa decemlineata ( L. decemlineata; Say) midgut and fat-body homogenate. In-vivo toxicity and the antifeedant activity were performed using larval bioassays. The methanol extracts had greater GST inhibitory activity compared to the glycoalkaloids, as well as greater 2nd instar larvae mortality and antifeedant activity. Furthermore, the green leaf volatile compound, cis-hex-3-enyl acetate, at the concentration of 5 ppm, caused 50% mortality of 2nd instar larvae. Our findings suggest the potential usefulness of S. nigrum and S. villosum extracts to control L. decemlineata.

Does Lycium europaeum leaf have antihyperglycemic, antihyperlipidemic and antioxidant effects

The purpose of the present investigation is to assess, for the first time, the antidiabetic, antihyperlipidemic and antioxidant activities of Lycium europaeum extract in alloxan-induced diabetic rats. Diabetes was induced in adult male Wistar rats via a single subcutaneous alloxan injection (120 mg/kg). Lycium europaeum aqueous extract was orally administered at a dose of 20 mg/kg for 28 consecutive days. Serum concentrations of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and triglycerides (TG) were assayed at the end of the experimental period in all investigated groups. Antioxidant enzymes such as glutathione peroxidase (GPx), catalase (CAT) and superoxide dismutase (SOD) were sought in the serum and pancreas. Lycium europaeum extract significantly increased HDL-C and reduced blood glucose, TC, LDL-C and TG as compared to the alloxan-control group. Lycium europaeum extract was also efficient in reducing oxidative stress in diabetic rats by increasing SOD, CAT and GPx activities both in the pancreas and the plasma of the animals. Moreover, Lycium europaeum extract contained considerable levels of polyphenols and flavonoids. It also exhibited an important antioxidant capacity and a remarkable ability to quench DPPH radicals and reduce irons. The obtained results highlight potentially relevant health beneficial effects of Lycium europaeum extract, reversing hyperglycemic, hyperlipidemic and oxidative stress effects in rats with alloxan-induced diabetes. Therefore, it may be considered as a promising alternative or complementary agent to diabetes treatment.

The phytochemical and bioactivity profiles of wild Asparagus albus L. plant.

The ethanolic extracts from the leaves, pericarps and rhizomes of Asparagus albus L. were investigated for their phytochemical composition, antioxidant (DPPH and FRAP assays), anti-microbial against human pathogenic isolates and cytotoxic (human colon carcinoma HCT-116 cells) activities. The highest flavonoid content was obtained in the leaf extract followed by the pericarp but there were no flavonoids detected in the rhizome. However, the rhizome had a high concentration of saponins. Flavonoid and saponin profiles were similar to those previously described for the triguero Huetor Tajar asparagus landrace. It was found that the pericarp ethanolic extract exhibited higher antioxidant activity than rhizome and leaf extracts. Moreover, the rhizome possessed more evident cytotoxic activity against HCT-116 cells in comparison to leaf and pericarp. All extracts showed varying degrees of antimicrobial activity against most of the human pathogenic isolates. In addition, the leaves showed more powerful inhibitory activities against the maximum number of bacteria and all the fungai isolated and the highest activity was in the pericarp extract against multidrug resistant Pseudomonas aeruginosa (MDR) and Erythromycin resistant Streptococcus agalactiae (ER) with an inhibition zone of 21mm and 19mm, respectively. The results show that A. albus could be a new crop with pharmaceutical interest because its richness in bioactive compounds provides considerable benefits for human health.

Decision-making criteria for plant-species selection for phytostabilization: Issues of biodiversity and functionality.

In polluted protected areas, using phytoremediation raises the question of the choice of the plant species to select. As an example, Atriplex halimus has been identified as a proliferative plant species that needs to be eradicated in the Calanques National Park (PNCal). Since it has been proven that the spontaneous populations of this plant species could phytostabilize shore waste deposits generated by past industrial activities within the PNCal territory, its status seems controversial, presenting a dilemma between biodiversity management of a protected area and ecological solutions for pollution management. To address this issue, we assessed the ability of A. halimus to grow on different soils from this territory, in order to estimate the potential invasiveness of this plant in this territory. Petri dish germinations and pot-growth experiments showed 50% germination of seeds collected on local individuals from the most polluted PNCal soil and 20% growth reduction of seedlings. Soil analysis showed that limitation of growth was caused by high pH value and sparsely available micronutrients as well as metal and metalloid contamination. Our results suggested that local populations of A. halimus may stabilize the highly metal and metalloid polluted salt-affected soils of the PNCal, with low seed germination potential lowering the eventuality of a propagation over the PNCal territory. As a consequence of this study, the administration of the PNCal decided not to remove A. halimus populations along the polluted coastline until another solution to prevent pollution dispersal had been found. This laboratory approach may be extended to other similar situations where plant species may be evaluated not only in term of phytoremediation potential but also in term of biodiversity preservation.

The interaction between foliar GA3 application and arbuscular mycorrhizal fungi inoculation improves growth in salinized tomato (Solanum lycopersicum L.) plants by modifying the hormonal balance.

The agriculture industry is frequently affected by various abiotic stresses limiting plant productivity. To decrease the negative effect of salinity and improve growth performance, some strategies have been used, such as exogenous application of plant growth regulators (i.e. gibberellic acid, GA3), or arbuscular mycorrhizal fungi (AMF) inoculation. To gain insights about the cross-talk effect of exogenous GA3 application and AMF inoculation on growth under salinity conditions, tomato plants (Solanum lycopersicum, cv. TT-115) were inoculated or not with the AMF Rhizophagus irregularis and exposed to different treatments during two weeks: 0M GA3+0mM NaCl, 10-6M GA3+0mM NaCl, 0M GA3+100mM NaCl and 10-6M GA3+100mM NaCl. Results have revealed that AMF inoculation or GA3 application alone, but especially their interaction, resulted in growth improvement under salinity conditions. The growth improvement observed in AMF-inoculated tomato plants under salinity conditions was mainly associated to ionic factors (higherK concentration and K/Na ratio) while the alleviating effect of GA3 application and its interaction with AMF appear to be due to changes in the hormonal balance. Foliar GA3 application was found to increase the active gibberellins (GAs), resulting in a positive correlation between GA3 and the growth-related parameters. Furthermore, cytokinins, indoleacetic acid and abscisic acid concentrations increased in AMF inoculated or GA3 treated plants but, notably, in AMF plants treated with GA3, which showed improved growth under salinity conditions. This suggests that there is an interactive positive effect between GAs and AMF which alleviates growth impairment under salinity conditions by modifying the hormonal balance of the plant.

Antioxidant and selective anticancer activities of two Euphorbia species in human acute myeloid leukemia.

In this study, two Euphorbia species (i.e. terracina and paralias) were investigated for their cytotoxic and antioxidant activities. Cytotoxicity of plant methanol and chloroform fractions was examined towards human acute myeloid leukemia (THP1) and human colon epithelial (Caco2) cancer cell lines, as well as CD 14 and IEC-6 normal cells by targeting various modulators of apoptosis or inflammation. Moreover, secondary metabolite pools (phenolic classes, alkaloids, terpenes, saponins) and antioxidant activities (DPPH, ABTS and O2- scavenging, as well as FRAP tests) were assessed in plant extracts. Both Euphorbia species appeared to be rich in phenolic compounds and terpenoids, Moreover, E. terracina polar and apolar fractions and E. paralias polar fraction were highly active against THP1 cells, with IC50 values of 2.08, 14.43 and 54.58µg/mL, respectively. However, no cytotoxicity was found against normal cells (CD14+ monocytes). The results indicate that the three fractions induce apoptosis in THP1 cell line after 6h of exposure. Furthermore, apoptosis caused by apolar fraction was related to a caspase-dependent process, whereas other death pathways seemed to be involved with the polar fractions. An enhanced production of reactive oxygen species was detected upon cell treatment with plant extracts. Interestingly, they have no effect on cytokine TNF-α secretion in THP1 and normal cells compared to untreated cells, indicating that the three fractions caused no inflammation. Euphorbia terracina and E. paralias polar fractions showed strong antioxidant activity with potent scavenging capacity against DPPH, ABTS and superoxide radicals. Moreover, these fractions displayed a very high ferric reducing power. These findings confirm the strong antioxidant capacity of Euphorbia plants and suggest a targeted anti-cancer effect with a potent anti-proliferative property of E. terracina and E. paralias extracts, which induce programmed cell death in leukemia cell lines but not in normal monocytes cells.

Phenolic content, antioxidant and anti-inflammatory activities of Tunisian Diplotaxis simplex (Brassicaceae).

This study investigates the polyphenol content of Diplotaxis simplex extract and the biological activities of the main organ. The analysed extracts showed that polyphenol contents varied considerably as a function of organs. Furthermore, novel biological activities of this species were assessed. Flower extracts exhibit a potent in vitro antioxidant capacity using oxygen radical absorbance capacity and displayed a strong anti-inflammatory activity, inhibiting nitric oxide release, by 79.3% at 160 µg/mL. Our findings suggested that the Diplotaxis flower is a valuable source of antioxidants and anti-inflammatory agents.

Salt stress mitigation by seed priming with UV-C in lettuce plants: growth, antioxidant activity and phenolic compounds.

Seeds of Lactuca sativa L. 'Romaine' were subjected to priming treatments with UV-C radiation at 0.85 or 3.42 kJ m(-2). Seedlings obtained from both primed (Pr) and non-primed (NPr) seeds were grown in an hydroponic culture system supplemented with 0 (control) or 100 mM NaCl. After 21 days of NaCl treatment, root and leaf biomass, root lengths, leaf numbers, and leaf surface area were measured. Ions (Na(+) and K(+)) accumulation was determined in roots and leaves. Total phenolic compound and flavonoid concentrations, as well as antioxidant and antiradical activities were measured in L. sativa leaves. Salt stress resulted in a lower increase in fresh weight of roots and leaves, which was more pronounced in roots than in leaves, due to reduced root elongation, leaf number and leaf expansion, as well as leaf thickness. The lower increase in fresh weight was accompanied by a restriction in tissue hydration and K(+) ion uptake, as well as an increase in Na(+) ion concentrations in all organs. These effects were mitigated in plants from the UV-C primed seeds. The mitigating effect of UV-C was more pronounced at 0.85 than at 3.42 kJ m(-2). Salt stress also resulted in an increase in total phenolic compounds and flavonoid concentrations and in the total antioxidant capacity in leaves. The highest diphenylpicrylhydrazyl radical (DPPH) scavenging activity was found in the leaves of plants from both Pr seeds. Our results suggest that plants grown from seed primed by exposure to moderate UV-C radiation exhibited a higher tolerance to salinity stress.

How does iron deficiency disrupt the electron flow in photosystem I of lettuce leaves?

The changes observed photosystem I activity of lettuce plants exposed to iron deficiency were investigated. Photooxidation/reduction kinetics of P700 monitored as ΔA820 in the presence and absence of electron transport inhibitors and acceptors demonstrated that deprivation in iron decreased the population of active photo-oxidizable P700. In the complete absence of iron, the addition of plant inhibitors (DCMU and MV) could not recover the full PSI activity owing to the abolition of a part of P700 centers. In leaves with total iron deprivation (0µM Fe), only 15% of photo-oxidizable P700 remained. In addition, iron deficiency appeared to affect the pool size of NADP(+) as shown by the decline in the magnitude of the first phase of the photooxidation kinetics of P700 by FR-light. Concomitantly, chlorophyll content gradually declined with the iron concentration added to culture medium. In addition, pronounced changes were found in chlorophyll fluorescence spectra. Also, the global fluorescence intensity was affected. The above changes led to an increased rate of cyclic electron transport around PSI mainly supported by stromal reductants.

Variability of phenolic content and antioxidant activity of two lettuce varieties under Fe deficiency.

BACKGROUND: Fe deficiency affects food growth and quality in calcareous soils. In this study, the effect of Fe deficiency on growth parameters, phenolic content and antioxidant capacities of two lettuce shoots varieties (Romaine and Vista) were investigated. RESULTS: Fresh matter production, pigment (chlorophyll and carotenoid) and Fe2+ content were significantly reduced by Fe deficiency in both varieties. However, restriction of these parameters was particularly pronounced in Romaine variety as compared to Vista. Moreover, Fe deficiency caused decreases in the activity of antioxidant enzymes such as catalase and guaiacol peroxidase, whereas ascorbate peroxidase and malondialdehyde concentrations were not significantly affected. On the other hand, Fe deficiency in Vista variety induced an increase in polyphenol and flavonoid content as compared to Romaine variety. In addition, total antioxidant capacity and antiradical test against DPPH radical decreased in leaves of Romaine variety after 15 days of treatment. CONCLUSION: These results suggest that the higher polyphenol content in Vista variety supports the involvement of these components in the stability of antioxidant capacities and then in its protection against oxidative damage generated by Fe deficiency in lettuce plants.

Salt effect on phenolics and antioxidant activities of Tunisian and Canadian sweet marjoram (Origanum majorana L.) shoots.

BACKGROUND: Two varieties of Origanum majorana (Canadian and Tunisian) were evaluated for their phenolic, flavonoid and tannin contents, individual phenolic compounds and antioxidant activities under NaCl constraint. RESULTS: The results showed a significant variability in phenolic composition and antioxidant behavior between the two varieties under salt stress. The phenolic composition of methanolic extracts was determined by reversed-phase high-performance liquid chromatography. Amentoflavone was the predominant flavonoid compound; in addition, trans-2-hydrocinnamic acid became the major phenolic acid with salt treatment of the Tunisian variety. In the control, Canadian variety extract was characterized by high levels of gallic acid and amentoflavone. However, under 75 mmol L(-1) NaCl, gallic acid content doubled, whereas amentoflavone content was maintained in the Canadian variety. Stimulation of phenolic acid biosynthesis was observed in these two varieties under salt treatment despite the fact that shoots of the Tunisian variety showed higher antioxidant activities compared to those from the Canadian variety. Tunisian O. majorana might have developed tolerance to salinity and avoided tissue damage by activating enzymes involved in the galactosylation of quercetin into quercetin-3-galactoside and quercetin-3-rhamnoside. CONCLUSION: Our results confirmed the tolerance of Tunisian O. majorana plants.

Salt effects on Origanum majorana fatty acid and essential oil composition.

BACKGROUND: The effects of salt on the essential oil yield and fatty acid composition of aerial parts of two marjoram varieties were investigated. Plants with 6 leaves were treated with NaCl (75mM). RESULTS: Salt treatment led to a reduction in aerial part growth. Salinity increased the fatty acid content more significantly in Tunisian variety (TV) than in Canadian variety (CV). CV showed an increase in double-bond index (DBI) and a decrease in malondialdehyde content under salt stress, while the opposite was observed in TV. The DBI was mainly affected by a strong reduction in oleic and linoleic acids in TV, whereas a strong stimulation of linoleic acid in CV was observed. Salt decreased and increased the essential oil yield in TV and CV respectively. The main constituents of the essential oil of TV were trans-hydrate sabinene and terpinen-4-ol, which showed a significant decrease under salt stress. In contrast, the main constituents of the essential oil of CV were sabinene and trans-hydrate sabinene, which showed a significant decrease and increase respectively under salt stress. CONCLUSION: Marjoram oil is a rich source of many compounds such as essential oils and fatty acids, but the distribution of these compounds differed significantly between the two varieties studied.

Inhibition of photosystems I and II activities in salt stress-exposed Fenugreek (Trigonella foenum graecum).

Fenugreek (Trigonella foenum graecum) seedlings were exposed to increasing NaCl concentrations in the growth medium to examine the effect of salt stress on the electron transport reactions of photosynthesis. Activities of both photosystem II (PSII), measured by chlorophyll fluorescence, and photosystem I (PSI), measured by P700 photooxidation, were decreased by salt stress. The inhibition proceeded in a two step manner. At the lower salt concentrations used and shorter exposition periods, electron transfer between the quinone acceptors of PSII, Q(A) and Q(B), was strongly retarded as shown by an increased amplitude of the OJ phase of the OJIP chlorophyll fluorescence induction traces and slowed chlorophyll fluorescence relaxation kinetics following a single turn-over flash. The above indicated a disturbance of the Q(B) binding site likely associated with the first step of photoinhibition. In the second step, strong photoinhibition was observed as manifested by increased F(0) values, declined F(v)/F(0) and loss of photoactive P700.

Inhibition of photosynthetic oxygen evolution and electron transfer from the quinone acceptor QA- to QB by iron deficiency.

The effect of iron deficiency on photosynthetic electron transport in Photosystem II (PS II) was studied in leaves and thylakoid membranes of lettuce (Lactuca sativa, Romaine variety) plants. PS II electron transport was characterized by oxygen evolution and chlorophyll fluorescence parameters. Iron deficiency in the culture medium was shown to affect water oxidation and the advancement of the S-states. A decrease of maximal quantum yield of PS II and an increase of fluorescence intensity at step J and I of OJIP kinetics were also observed. Thermoluminescence measurements revealed that charge recombination between the quinone acceptor of PS II, Q(B), and the S(2) state of the Mn-cluster was strongly perturbed. Also the dark decay of Chl fluorescence after a single turnover white flash was greatly retarded indicating a slower rate of Q(A)(-) reoxidation.

The impact of genotype and salinity on physiological function, secondary metabolite accumulation, and antioxidative responses in lettuce.

Salinity inhibits plant growth due to osmotic and ionic effects. However, little is known about the impact of genotype and salinity on biochemical and molecular processes in the leafy vegetable lettuce. We report here evaluations of two lettuce types, Verte (NaCl tolerant) and Romaine (NaCl sensitive), under iso-osmotic 100 mM NaCl and 77 mM Na(2)SO(4) treatments. As compared to Romaine, NaCl-treated Verte displayed better growth, contained lower levels of inorganic cations in leaves, and possessed superior antioxidative capacity due to enhanced carotenoid and phenolics biosynthesis and more active antioxidative enzymes resulting in reduced membrane damage. Both genotypes had relatively similar growth patterns under Na(2)SO(4) treatment, but Romaine showed enhanced root lignification, greater malondialdehyde formation, and suppressed Fe-superoxide dismutase expression in roots as compared with Verte.

Comparative effect of potassium on K and Na uptake and transport in two accessions of Arabidopsis thaliana during salinity stress.

Potassium-sodium interaction was compared in two natural accessions of Arabidopsis thaliana, Columbia-0 and NOK2. Seedlings were grown in the presence of 0 or 50 mM NaCl and 0.1; 0.625 or 2.5 mM K(+). At the lowest K(+) concentration, salt treatment inhibited both K(+) uptake and growth. Increasing the K(+) availability did not modified salt response in Columbia-0, but restored nearly normal net K(+) uptake in NaCl condition and alleviated NaCl growth reduction in NOK2. The effect of K(+) and NaCl on transcript level of several K(+) and Na(+) transporters in both shoots and roots was assessed using semi-quantitative RT-PCR. The mRNA abundance of the NHX1 and SOS1 Na(+)/H(+) antiporters was significantly increased by 50 mM NaCl in the two accessions. NHX1, which is responsible for Na(+) sequestration into vacuoles, was more up-regulated in NOK2 leaves than in Columbia-0's in NaCl stress condition. AKT1, which is the major channel involved in K(+) absorption, was down-regulated in salt stress condition, but was not responding to K(+) treatments. Only in NOK2, SKOR and AKT2, which respectively control xylem and phloem K(+) transport, were markedly up-regulated by 2.5 mM K(+) in both roots and shoots, independently of NaCl. Phenotypic and gene expression analyses suggest that the relative salt tolerance of NOK2 is mainly due to a high ability to sequester Na(+) in the vacuole and to take up and transport K(+). Up-regulation of SKOR and AKT2 by K(+), and of NHX1 by NaCl could participate in determining this phenotype.

Long-term effects of mild salt stress on growth, ion accumulation and superoxide dismutase expression of Arabidopsis rosette leaves.

Arabidopsis thaliana plants (wild-type accessions Col and N1438) were submitted to a prolonged, mild salt stress using two types of protocols. These protocols allowed salt-treated plants to absorb nutrients either through a part of their root system maintained in control medium (split-rooted plants) or during episodes on control medium alternating with salt application (salt alternation experiment). Full-salt treatments (salt applied continuously to whole root system) resulted in severe (but non-lethal) growth inhibition. This effect was partly alleviated in split-rooted plants on mixed salt-control medium and in plants submitted to salt-control medium alternation. The activity of the various isoforms of superoxide dismutases (SODs) did not appreciably change with the treatments. The abundance of the mRNAs of the seven SOD genes present in Arabidopsis genome was determined using real-time polymerase chain reaction. The two protocols gave qualitatively identical results. The expression level was increased by full-salt treatments for some genes and diminished for other genes. However, the nature of these genes differed according to the accessions: the responses to salt of FSD1 and MSD were opposite in Col and N1438. In Col, salt treatments inhibited the expression of FSD1 and strongly stimulated that of CSD1 and MSD. In N1438, the stimulation by salt concerned FSD1 and CSD1 and MSD expression being inhibited. In both accessions, the expression of CSD2 and CSD3 was lowered by salt. For all genes, the treatments that mitigated stress partially restored SOD expression to control level. Thus, the changes in SOD transcript abundance accurately reflected the severity of the salt stress.

Phenotypic and molecular characterization of chickpea rhizobia isolated from different areas of Tunisia.

Several phenotypic markers were used in this study to determine the biodiversity of rhizobial strains nodulating Cicer arietinum L. in various areas of Tunisia. They include symbiotic traits, the use of 21 biochemical substrates, and tolerance to salinity and pH. In addition, restriction fragment length polymorphisms (RFLPs) of PCR-amplified 16S rDNA were compared with those of reference strains. Numeric analysis of the phenotypic characteristics showed that the 48 strains studied fell into three distinct groups. This heterogeneity was highly supported by the RFLP analysis of 16S rRNA genes, and two ribotypes were identified. Chickpea rhizobia isolated from Tunisian soils are both phenotypically and genetically diverse. Results showed that 40 and 8 isolates were assigned, respectively, to Mesorhizobium ciceri and Mesorhizobium mediterraneum.