Isolation of the PCB-degrading bacteria Mesorhizobium sp. ZY1 and its combined remediation with Astragalus sinicus L. for contaminated soil

A bacterial strain ZY1 capable of utilizing PCBs as its carbon source was isolated from the root nodules of Chinese milk vetch (Astragalus sinicus L.). The strain was identified as Mesorhizobium sp. according to its physiological-biochemical properties and the analysis of its 16S rRNA gene sequence. When the initial OD600 was 0.15, 62.7% of 15 mg L^?1 3,3′,4,4′-TCB in a liquid culture was degraded by Mesorhizobium sp. ZY1 within 10 days. Mesorhizobium sp. ZY1 also greatly increased the biotransformation of soil PCBs. Pot experiments indicated that the soil PCB concentrations of a single incubation of strain ZY1 (R) and a single planting of A. sinicus (P) decreased by 20.5% and 23.0%, respectively, and the concentration of PCBs in soil treated with A. sinicus and strain ZY1 decreased by 53.1%. We also observed that A. sinicus-Mesorhizobium sp. ZY1 treatment (PR) improved plant biomass and the concentration of PCBs in plants compared with a single A. sinicus planting treatment (P). The results suggest that the synergistic association between A. sinicus and PCBs-degrading Mesorhizobium sp. ZY1 can stimulate the phytoextraction of PCBs and the rhizosphere microflora to degrade PCBs, and might be a promising bioremediation strategy for PCB-contaminated soil.     

Assessment of AM biotechnology in improving agricultural productivity of nutrient-deficient soil in the tropics

Abstract

Growth, nutrient content and nodulation response of cowpea plants (Vigna unguiculata L. Walp) inoculated with a Arbuscular Mycorrhiza (AM) fungus (Glomus etunicatum) and Bradyrhizobium (BR) strain IRC 25B peat-based were assessed on an alfisol in a two-cropping cycle experiment conducted in the greenhouse. A total of 5 kg sieved unsterilized topsoil plastic pots was amended with compost consisting of 2.4% N, 1.7% P, 2.7% Ca, 0.4% Mg and 0.7% Fe. Analysis from this first cropping cycle showed that all cowpea plants were infected with mycorrhiza in both AM inoculated and uninoculated treatments. However, all the AM inoculated plants had higher infections than the uninoculated cowpea plants. Nevertheless, nodule number and nodule weight of cowpea plants generally increased in response to compost application when used alone, or when combined with BR or AM; except for nodule weight of BR + Compost treatment. At 13 weeks after planting, the plants were harvested for a second cropping cycle experimental analysis. Results showed higher mycorrhizal infections in all the treatments inoculated with AM. However, infection was highest in cowpea plants treated with AM + BR + Compost, followed by those treated with AM + BR. This shows an increase in the number of AM propagules during the period of cropping. All other parameters measured were found generally lower in their mean values compared to the first cropping cycle. It was observed in this study that compost applications with AM inoculation could substitute for inorganic fertilizer. Thus, tropical countries should direct their efforts towards making the best use of AM to improve conditions for the peasant farmers that account for over 70% agricultural productivity in the region.     

Competition for Astragalus sinicus root nodule infection between its native microsymbiont Rhizobium huakuii bv. renge B3 and Rhizobium sp. ACMP18 strain, specific for Astragalus cicer

Rhizobium huakuii bv. renge B3, a native symbiont of Astragalus sinicus, outcompeted Rhizobium sp. strain ACMP18, which was isolated from Astragalus cicer nodules, in the formation of root nodules on A.␣sinicus when plants were co-inoculated with these strains. The strains occupying the nodules were identified by antibiotic resistance and phage sensitivity markers and also by polymerase chain reaction (PCR) genomic fingerprintings, which were performed by using enterobacterial repetitive intergenic consensus sequences. In PCR genomic fingerprintings, the total genomic DNA isolated from pure bacterial culture and from squashed root nodules showed identical profiles, indicating that this technique can be a useful tool for identification of rhizobia in ecological studies. When Rhizobium sp. strain ACMP18 outnumbered R. huakuii bv. renge strain B3 by a factor of ten, and even when strain ACMP18 was added to plants 1 week before bacterization with strain B3, the strain B3 occupied most nodules. Dually infected nodules were not observed, although Rhizobium sp. ACMP18 formed active nodules on A. sinicus when the bacterial strain was inoculated alone. Received: 5 February 1998 / Received revision: 23 March 1998 / Accepted: 27 April 1998     

A novel method to alleviate arsenic toxicity in alfalfa plants using a deletion mutant strain of Sinorhizobium meliloti

Reduction in crop yield and contamination of food crops are major problems in many areas due to high soil arsenic content. In this study an aquaglyceroporin (AqpS) disrupted Sinorhizobium meliloti smk956 strain was found to accumulate 70.5% more arsenic than its parental strain S. meliloti Rm1021 under free living condition. This strain was inoculated onto alfalfa host plants under different arsenic concentrations (0, 1 and 5 mg/L) and its ability to alleviate arsenic toxicity in the host plant was investigated. At 1 and 5 mg/L arsenic concentrations the average arsenic contents in the shoots of the plants inoculated with the strain S. meliloti smk956 were 45.5 and 27.5% less than those of the plants inoculated with S. meliloti Rm1021, respectively. Under arsenic stress conditions the strain S. meliloti smk956 showed increased symbiotic efficiency than its parental strain. These results demonstrate a novel method to alleviate arsenic toxicity in alfalfa plants.     

Expression of the Kidney Bean Phenylalanine-Ammonia Lyase Gene in the Hairy Roots of Astragalus sinicus

Using Agrobacterium rhizogenes, Astragalus sinicus plants were transformed with the kidney bean pal5 gene coding for phenylalanine-ammonia lyase (PAL). The hairy root culture thus obtained manifested enhanced PAL activity and lignin content in the cell walls; in addition, the transformed cells differed from the wild-type ones in several electrophysiological indices. In particular, the diffusion component of the total membrane potential of plasmalemma increased in the pal-transformed roots. The authors presume that the volume density of the protein-related negative charge of the cytoplasm increases in the transformed root cells along with changes in the cytoplasmic pH and pCa²⁺, the extent of coupling of these two indices, and the hydraulic conductivity of plasmodesmata.     

Enhancement of the efficiency of Cd phytoextraction using bacterial endophytes isolated from Chromolaena odorata,a Cd hyperaccumulator

Phytoextraction is a technique using a hyperaccumulator to remove heavy metals from soil. The efficiency of heavy metal uptake can be enhanced by the inoculation of endophytes. In this study, we isolated and identified 23 endophytes from Chromolaena odorata, a cadmium (Cd) hyperaccumulator that consisted of 19 bacteria, 2 actinomycetes and 2 fungi. All bacteria and fungi could produce at least 1 plant growth promoting factors. However, only 4 bacterial isolates; Paenibacillus sp. SB12, Bacillus sp. SB31, Bacillus sp. LB51, and Alcaligenes sp. RB54 showed the highest minimum inhibitory concentration (MIC) value (2.9 mM), followed by Exiguobacterium sp.RB51 (1.7 mM). Then, these 5 high-MIC bacteria and 1 low-MIC bacterium, Bacillus sp. LB15 were inoculated onto sunflower grown in soil supplemented with 250 mg/kg of Cd. After 60 days, all inoculated plants accumulated significantly higher Cd concentration than the non-inoculated counterparts, and those inoculated with strain LB51 showed the highest Cd accumulation and growth. Interestingly, strain LB15 with low MIC also enhanced Cd accumulation in plants. The results suggest that these bacteria, particularly strain LB51, could be applied to improve Cd accumulation in plants, and that bacteria with low MIC also have the potential to enhance the efficiency of phytoextraction.     

Exiguobacterium oxidotolerans, a halotolerant plant growth promoting rhizobacteria, improves yield and content of secondary metabolites in Bacopa monnieri (L.) Pennell under primary and secondary salt stress

Brahmi (Bacopa monnieri), an integral component of Indian Ayurvedic medicine system, is facing a threat of extinction owing to the depletion of its natural populations. The present study investigates the prospective of exploitation of halotolerant plant growth promoting rhizobacteria (PGPR) in utilising the salt stressed soils for cultivation of B. monnieri. The effects of two salt tolerant PGPR, Bacillus pumilus (STR2) and Exiguobacterium oxidotolerans (STR36) on the growth and content of bacoside-A, an important pharmaceutical compound in B. monnieri, were investigated under primary and secondary salinity conditions. The herb yields of un-inoculated plants decreased by 48 % under secondary salinization and 60 % under primary salinization than the non salinised plants. Among the rhizobacteria treated plants, E. oxidotolerans recorded 109 and 138 %, higher herb yield than non-inoculated plants subjected to primary and secondary salinity respectively. E. oxidotolerans inoculated plants recorded 36 and 76 % higher bacoside-A content under primary and secondary salinity respectively. Higher levels of proline content and considerably lower levels of lipid peroxidation were noticed when the plants were inoculated with PGPR under all salinity regimes. From the results of this investigation, it can be concluded that, the treatments with salt tolerant PGPR can be a useful strategy in the enhancement of biomass yield and saponin contents in B. monnieri, as besides being an eco-friendly approach; it can also be instrumental in cultivation of B. monnieri in salt stressed environments.     

Contribution of native phosphorous-solubilizing bacteria of acid soils on phosphorous acquisition in peanut (<Emphasis Type="Italic">Arachis hypogaea</Emphasis> L.)

The present investigation analyzes the in vitro P solubilization [Ca-P, Al-P, Fe(II)-P, and Fe(III)-P] efficiency of native PSB strains from acid soils of Odisha and exploitation of the same through biofertilization in peanut (Arachis hypogaea L.) growth and P acquisition. One hundred six numbers of soil samples with pH ≤ 5.50 were collected from five districts of Odisha viz., Balasore, Cuttack, Khordha, Keonjhar, and Mayurbhanj. One bacterial isolate from each district were selected and analyzed for their P solubilization efficiency in National Botanical Research Institute Phosphate broths with Ca, Al, and Fe-complexed phosphates. CTC12 and KHD08 transformed more amount of soluble P from Ca-P (CTC12 393.30 mg/L; KHD08 465.25 mg/L), Al-P (CTC12 40.00 mg/L; KHD08 34.50 mg/L), Fe(III)-P (CTC12 175.50 mg/L; KHD08 168.75 mg/L), and Fe(II)-P (CTC12 47.40 mg/L; KHD08 42.00 mg/L) after 8 days of incubation. The bioconversion of P by all the five strains in the broth medium followed the order Ca-P > Fe(III)-P > Fe(II)-P > Al-P. The identified five strains were Bacillus cereus BLS18 (KT582541), Bacillus amyloliquefaciens CTC12 (KT633845), Burkholderia cepacia KHD08 (KT717633), B. cepacia KJR03 (KT717634), and B. cepacia K1 (KM030037) and further studied for biofertilization effects on peanut. CTC12 and KHD08 enhanced the soil available P around 65 and 58% and reduced the amount of each Al³⁺ about 79 and 81%, respectively, over the uninoculated control pots in the peanut rhizosphere. Moreover, all tested PSB strains could be able to successfully mobilize P from inorganic P fractions (non-occluded Al-P and Fe-P). The strains CTC12 and KHD08 increased the pod yield (114 and 113%), shoot P (92 and 94%), and kernel P (100 and 101%), respectively, over the control. However, B. amyloliquefaciens CTC12 and B. cepacia KHD08 proved to be the potent P solubilizers in promoting peanut growth and yield.     

Application of Bacillus megaterium MCR-8 improved phytoextraction and stress alleviation of nickel in Vinca rosea

The current research was performed to evaluate the effect of Bacillus megaterium MCR-8 on mitigation of nickel (Ni) stress in Vinca rosea grown on Ni-contaminated soil (50, 100, and 200 mg Ni kg^?1 soil). The treated plants exhibited reduced growth, biomass, gas exchange capacity, and chlorophyll (Chl) content under Ni stress. The inoculated plants growing in Ni-contaminated media exhibited relatively higher growth, total soluble protein, and proline contents. Similarly, bacterial inoculation improved the activity of antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and ascorbate peroxidase (APX) under Ni stress. The Ni stress alleviation in inoculated plants was attributed to the reduced level of malondialdehyde (MDA) and hydrogen peroxide (H₂O₂), enhanced synthesis of protein, proline, phenols, and flavonides in conjunction with improved activity of antioxidant enzymes. The growth-promoting characteristics of microbe such as 1-aminocyclopropane-1-carboxylate deaminase (ACCD) and phosphate solubilization activity, siderophore, and auxin production capability also improved the growth and stress mitigation in inoculated plants. Furthermore, the inoculated plants exhibited higher value for bioconcentration factor (BCF), translocation factor (TF), and resulted in higher loss of Ni content from soil. The current results exhibited the beneficial role of B. megaterium MCR-8 regarding stress alleviation and Ni phytoextraction by V. rosea.     

Characterization ofBradyrhizobium sp. (Astragalus sinicus L.) using serological agglutination,intrinsic antibiotic resistance,plasmid visualization,and field performance

Eleven isolates ofBradyrhizobium sp. (Astragalus sinicus L.), originating from plants ofAstragalus sinicus (Chinese milkvetch) in China and Japan, were characterized using serological agglutination, intrinsic antibiotic resistance, and plasmid visualization. Serological agglutination, using antisera against three isolates effective onA. sinicus and two isolates effective on otherAstragalus spp., showed that there may be two serogroups among the eleven isolates effective onA. sinicus, and that the antigenic components are heterogeneous. Results of both intrinsic antibiotic resistance and plasmid visualization demonstrated similarities between four of the isolates.Ten isolates were further evaluated for effectiveness on the host,A. sinicus, in a field study in Puyallup, Washington (47°N, 122°W). This study showed that the Chinese commercial inoculum and one field isolate were about ten times more effective than the control based on plant dry weight and seed yield. Eight other isolates were less effective.     

Regulation of plasma membrane aquaporins by inoculation with a Bacillus megaterium strain in maize (Zea mays L.) plants under unstressed and salt-stressed conditions

It is documented that some plant-growth-promoting rhizobacteria (PGPR) enhance plant salt tolerance. However, as to how PGPR may influence two crucial components of plant salt tolerance such as, root hydraulic characteristics and aquaporin regulation has been almost unexplored. Here, maize (Zea mays L.) plants were inoculated with a Bacillus megaterium strain previously isolated from a degraded soil and characterized as PGPR. Inoculated plants were found to exhibit higher root hydraulic conductance (L) values under both unstressed and salt-stressed conditions. These higher L values in inoculated plants correlated with higher plasma membrane type two (PIP2) aquaporin amount in their roots under salt-stressed conditions. Also, ZmPIP1;1 protein amount under salt-stressed conditions was higher in inoculated leaves than in non-inoculated ones. Hence, the different regulation of PIP aquaporin expression and abundance by the inoculation with the B. megaterium strain could be one of the causes of the different salt response in terms of root growth, necrotic leaf area, leaf relative water content and L by the inoculation treatment.     

Biosafety and colonization of Burkholderia multivorans WS-FJ9 and its growth-promoting effects on poplars

Burkholderia cepacia complex (Bcc) is a group of bacteria with conflicting biological characteristics, which make them simultaneously beneficial and harmful to humans. They have been exploited for biocontrol, bioremediation, and plant growth promotion. However, their capacity as opportunistic bacteria that infect humans restricts their biotechnological applications. Therefore, the risks of using these bacteria should be assessed. In this study, Burkholderia multivorans WS-FJ9 originally isolated from pine rhizosphere, which was shown to be efficient in solubilizing phosphate, was evaluated with respect to its biosafety, colonization in poplar rhizosphere, and growth-promoting effects on poplar seedlings. Pathogenicity of B. multivorans WS-FJ9 on plants was determined experimentally using onion and tobacco as model plants. Onion bulb inoculated with B. multivorans WS-FJ9 showed slight hypersensitive responses around the inoculation points, but effects were not detectable based on the inner color and odor of the onion. Tobacco leaves inoculated with B. multivorans WS-FJ9 exhibited slightly water-soaked spots around the inoculation points, which did not expand or develop into lesions even with repeated incubation. Pathogenicity of the strain in alfalfa, which has been suggested as an alternative Bcc model for mice, was not detectable. Results from gene-specific polymerase chain reactions showed that the tested B. multivorans WS-FJ9 strain did not possess the BCESM and cblA virulence genes. Scanning electron microscopy revealed that the colonization of the WS-FJ9 strain reached 1.4?×?10⁴ colony forming units (cfu)?g^?1 rhizosphere soil on day 77 post-inoculation. The B. multivorans WS-FJ9 strain could colonize the rhizosphere as well as the root tissues and cells of poplars. Greenhouse evaluations in both sterilized and non-sterilized soils indicated that B. multivorans WS-FJ9 significantly promoted growth in height, root collar diameter, and plant biomass of inoculated poplar seedlings compared with controls. Phosphorus contents of roots and stems of treated seedlings were 0.57 and 0.55 mg g^?1 higher than those of the controls, respectively. Phosphorus content was lower in the rhizosphere soils by an average of 1.03 mg g^?1 compared with controls. The results demonstrated that B. multivorans WS-FJ9 is a nonpathogenic strain that could colonize the roots and significantly promote the growth of poplar seedlings.     

The Effect of Recombinant Heavy Metal-Resistant Endophytic Bacteria on Heavy Metal Uptake by Their Host Plant

The ncc-nre nickel resistance system of Ralstonia metallidurans 31A was efficiently expressed in Burkholderia cepacia L.S.2.4 and Herbaspirillum seropedicae LMG2284. The heterologous expression of ncc-nre encoded nickel resistance was accompanied by nickel removal from the culture medium. B. cepacia L.S.2.4:: ncc-nre and H. seropedicae LMG2284::ncc-nre were able to remove 35 and 15% nickel, respectively. The capacity to remove nickel through sequestration or bio-precipitation processes and consequently lowering the free nickel concentration could offer interesting benefits for these endophytic bacteria and their host plants. Once inoculated in their host plant, they could possibly alter the nickel speciation and therefore decrease the free ions and thus toxic concentration for the plant metabolism. Lupinus luteus L, when grown on a nickel enriched substrate and inoculated with B. cepacia L.S.2.4::ncc-nre, showed a significant increase (30%) of nickel concentration in the roots, whereas the nickel concentration in the shoots remained comparable with that of the control plants. The inoculation of Lolium perenne (cv Atlas) with the nickel resistance derivative of H. seropedicae LMG2284::ncc-nre resulted in a significant decrease of the nickel concentration in the roots (11%) as well as in the shoots (14%). As this phenomenon was also observed in the Lolium perenne plants inoculated with the wild-type strain LMG2284, the nickel resistance characteristics probably are not responsible for the altered nickel uptake observed.     

Medicago sativa - Sinorhizobium meliloti Symbiosis Promotes the Bioaccumulation of Zinc in Nodulated Roots

In this study we investigated effects of Zn supply on germination, growth, inorganic solutes (Zn, Ca, Fe, and Mg) partitioning and nodulation of Medicago sativa This plant was cultivated with and without Zn (2 mM). Treatments were plants without (control) and with Zn tolerant strain (S532), Zn intolerant strain (S112) and 2 mM urea nitrogen fertilisation. Results showed that M. sativa germinates at rates of 50% at 2 mM Zn. For plants given nitrogen fertilisation, Zn increased plant biomass production. When grown with symbionts, Zn supply had no effect on nodulation. Moreover, plants with S112 showed a decrease of shoot and roots biomasses. However, in symbiosis with S532, an increase of roots biomass was observed. Plants in symbiosis with S. meliloti accumulated more Zn in their roots than nitrogen fertilised plants. Zn supply results in an increase of Ca concentration in roots of fertilised nitrogen plants. However, under Zn supply, Fe concentration decreased in roots and increased in nodules of plants with S112. Zn supply showed contrasting effects on Mg concentrations for plants with nitrogen fertilisation (increase) and plants with S112 (decrease). The capacity of M. sativa to accumulate Zn in their nodulated roots encouraged its use in phytostabilisation processes.     

ACC Deaminase-Containing Bacillus subtilis Reduces Stress Ethylene-Induced Damage and Improves Mycorrhizal Colonization and Rhizobial Nodulation in Trigonella foenum-graecum Under Drought Stress

This study was aimed at protecting Trigonella plants by reducing stress ethylene levels through ACC (1-aminocyclopropane-1-carboxylic acid) deaminase-containing Bacillus subtilis (LDR2) and promoting plant growth through improved colonization of beneficial microbes like Ensifer meliloti (Em) and Rhizophagus irregularis (Ri) under drought stress. A plant growth-promoting rhizobacterium strain possessing high levels of ACC deaminase characterized as B. subtilis was selected. Application of this strain considerably protected Trigonella plants under severe drought stress conditions; this protection was correlated with reduced levels of ACC (responsible for generation of stress ethylene). The experiment consisted of eight inoculation treatments with different combinations of ACC deaminase-containing rhizobacteria LDR2, Ri, and Em under three water regimes. The tripartite combination of LDR2 + Ri + Em acted synergistically to induce protective mechanisms against decreased soil water availability in Trigonella plants and improved plant weight by 56 % with lower ACC concentration (39 % less than stressed noninoculated plants) under severe drought conditions. Drought-induced changes in biochemical markers like reduced chlorophyll concentration, increased proline content, and higher lipid peroxidation were monitored and clearly indicated the protective effects of LDR2 under drought stress. Under drought conditions, apart from alleviating ethylene-induced damage, LDR2 enhanced nodulation and arbuscular mycorrhizal fungi colonization in the plants resulting in improved nutrient uptake and plant growth.     

Influence of arbuscular mycorrhiza on the growth and antioxidative activity in cyclamen under heat stress

The influence of the arbuscular mycorrhizal (AM) fungus, Glomus fasciculatum, on the growth, heat stress responses and the antioxidative activity in cyclamen (Cyclamen persicum Mill.) plants was studied. Cyclamen plants (inoculated or not with the AM fungus) were placed in a commercial potting media at 17–20 °C for 12 weeks in a greenhouse and subsequently subjected to two temperature conditions in a growth chamber. Initially, plants were grown at 20 °C for 4 weeks as a no heat stress (HS?) condition, followed by 30 °C for another 4 weeks as a heat stress (HS+) condition. Different morphological and physiological growth parameters were compared between G. fasciculatum-inoculated and noninoculated plants. The mycorrhizal symbiosis markedly enhanced biomass production and HS + responses in plants compared to that in the controls. A severe rate of leaf browning (80–100 %) was observed in control plants, whereas the mycorrhizal plants showed a minimum rate of leaf browning under HS + conditions. The mycorrhizal plants showed an increase activity of antioxidative enzymes such as superoxide dismutase and ascorbate peroxidase, as well as an increase in ascorbic acid and polyphenol contents. The 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity also showed a greater response in mycorrhizal plants than in the control plants under each temperature condition. The results indicate that in cyclamen plants, AM fungal colonisation alleviated heat stress damage through an increased antioxidative activity and that the mycorrhizal symbiosis strongly enhanced temperature stress tolerance which promoted plant growth and increased the host biomass under heat stress.     

Increased photosyntethic efficiency generated by fungal symbiosis in Agave victoria-reginae

A unique non-pathogenic strain of Fusarium oxysporum was isolated from field-collected plants of Agave victoria-reginae Moore. It was then re-introduced into micropropagated plants of the same species under greenhouse conditions. The fungic inoculation induced a 225% increase in the length of roots, 50% in the number of root branches and 50% in the number of stomata on the adaxial surface of leaves. Also, an increment of 167% in nocturnal acidity and a 122% in malic acid was observed and nocturnal pH was significantly more acid in the inoculated plants. Total chlorophyll and sugar content increased 14 and 172%, respectively. These results indicate a higher photosynthetic efficiency of the plants inoculated with the fungus than those plants which were not inoculated; therefore the association of this unique F. oxysporum strain with A. victoria-reginae Moore was considered as a beneficial symbiosis.     

Oxidative stress in the root nodules of Phaseolus vulgaris is induced under conditions of phosphorus deficiency

One of the most adverse effects of phosphorus (P) deficiency on N₂-fixing legumes is the generation of harmful active oxygen species which cause oxidative stress. And although oxidative stress has been widely studied in roots and shoots of various plant species, it has not yet sufficiently been documented in bean nodules so far. In this study, two recombinant inbred lines RIL115 (P-deficiency tolerant) and RIL147 (P-deficiency sensitive) of common bean and Concesa (local variety) were inoculated separately with the reference strain R. tropici CIAT899, RhM₁₁ (R. gallicum) or RhM₁₄ (R. tropici); two local strains of the Marrakesh region of Morocco. Nodulated plants were grown under semi-hydroponic conditions with sufficient or deficient P supply and analyzed for their oxidative responses at the flowering stage. The results indicated that P-deficiency decreased the growth of shoots (48 %) and nodules (32 %), particularly with RhM₁₄ exhibiting the highest decrease (52 %) of nodulation. This constraint increased electrolyte leakage of nodules (40 %) as compared to leaves (20 %), especially for plants inoculated with RhM₁₄ and CIAT899. Moreover, high H₂O₂ and malondialdehyde contents were noticed in P-deficient nodules of RhM₁₄ and RhM₁₁. These variations were associated with peroxidase activity stimulation in P-deficient nodules induced by CIAT899 and RhM₁₄. In symbiosis with RIL115, these last strains exhibited the highest nodule phenol content. Overall, phenol content was mainly enhanced in P-deficient nodules (35 %) as compared to the leaves (16 %). It was concluded that the genotypes inoculated with CIAT899 and RhM₁₁ are relatively P-deficiency tolerant combinations as compared to those inoculated with RhM₁₄. Increase of oxidative stress in nodules rather than in leaves points to the need for further investigations of mechanisms that improve the root-nodule efficiency under adverse conditions.     

Phenotypic and Phylogenetic Characterization of Burkholderia (Pseudomonas) sp. Strain LB400

The relevant phenotypic traits and phylogenetic relationships between Burkholderia (Pseudomonas) sp. strain LB400 and B. cepacia ATCC 25416^T were compared to determine the degree to which these two strains might be related. Strain LB400 degrades chlorinated biphenyls and has been a model system for potential use in the bioremediation of polychlorinated biphenyls, while some strains of B. cepacia are plant and human pathogens. The fatty acid methyl ester profile, sole carbon source utilization, and biochemical tests confirmed that strain LB400 was a member of the genus Burkholderia. The 16S rRNA gene sequence showed that this strain was not as closely related to B. cepacia as previously suspected or to other known pathogens of this genus, but is closely related to B. phenazinium, B. caribensis, B. graminis, and three unnamed Burkholderia spp. not known to be pathogenic. Received: 16 August 2000 / Accepted: 27 September 2000