Non-pathogenic Fusarium strains protect the seedlings of Lepidium sativum from Pythium ultimum

Two root-colonizing Fusarium strains, Ls-F-in-4-1 and Rs-F-in-11, isolated from roots of Brassicaceae plants, induced the resistance in Lepidium sativum seedlings against Pythium ultimum. These strains caused an increase in the content of benzyl isothiocyanate, and of its precursor glucotropaeolin, in the roots of the host plants. The increased isothiocyanate content is one of the factors contributing to the resistance of L. sativum against P. ultimum. To be transformed into the fungitoxic compound benzyl isothiocyanate, glucotropaeolin has to be hydrolyzed by myrosinase, which can be produced either by plants or microorganisms. The Fusarium strain Ls-F-in-4-1 has a myrosinase activity but the strain Rs-F-in-11 has not. These results suggest that both strains are able to trigger the metabolic pathway leading to benzyl isothiocyanate production in the plant. In the case of the myrosinase-negative strain Rs-F-in-11, hydrolyzation into isothiocyanate is only due to the myrosinase activity of the plant, and in the other case, the myrosinase produced by the strain Ls-F-in-11 also would contribute to the production of isothiocyanate. This paper reports a new mode of action of non-pathogenic Fusarium strains in controlling P. ultimum.     

Symbiotic effectiveness of Bradyrhizobium japonicum in acid soils can be predicted from their sensitivity to acid soil stress factors in acidic agar media

In acid soil, low pH, reduced availability of nutrients, and toxicity of Al and Mn limit plant growth and the survival and effectiveness of rhizobia. The symbiosis between legumes and rhizobia is particularly sensitive to acid soil stress. A pot experiment evaluated whether Bradyrhizobium japonicum strain growth on acidic agar media would predict ability to colonize the rhizosphere and form effective nodules in acidic soils. Three Indonesian strains of B. japonicum with similar effectiveness at neutral pH in sand culture but with different tolerance of acid soil stress factors in agar media, and an acid-tolerant commercial strain (CB1809) of comparable effectiveness, were tested in three acid soils using the Al tolerant soybean (Glycine max cv PI 416937). At 7 days after inoculation all strains had achieved large rhizosphere populations, but by day 14 the rhizosphere population of the acid-sensitive strain had decreased, while the more acid-tolerant strains increased. The acid-tolerant strains had significantly greater nodulation and symbiotic effectiveness than plants inoculated with the acid-sensitive strain. Laboratory prescreening of B. japonicum for acid, Al and Mn tolerance in acid media successfully identified strains which were symbiotically competent in low pH soils.     

Influence of soil on the interactions between endomycorrhizae and Azospirillum in sorghum

Sorghum (S. bicolor L. Moench cv. Bok 8) plants were grown in soil or sand-perlite low in plant-available N and P. Plants were inoculated with a vesicular-arbuscular mycorrhizal (VAM) fungus, or a strain of Azospirillum brasilense or both endophytes together. Plants received a nutrient solution which did not contain N or P. Increases in plant dry weight, shoot-to-root ratios, and the N content of dually-infected plants could be accounted for by summing the VAM and Azospirillum effects. For sorghum inoculated with both endophytes, the presence of A. brasilense in the rhizosphere increased VAM colonization and biomass, while the N input due to Azospirillum decreased, possibly due to competition for carbohydrates.Comparisons between sorghum grown with or without VAM-fungal infection in four growth media showed that edaphic factors other than P availability determined the host response to VAM infection. The P-fixing capacity of the soil, rather than the amount of available (NaHCO₃-extractable) P, influenced the balance between mutualistic and parasitic VAM-fungal growth.     

Inoculation of field-grown wheat (Triticum aestivum) with Azospirillum spp. in Brazil

Summary Three field experiments with wheat were conducted in 1983, 1984, and 1985 in Terra Roxa soil in Paraná, the major Brazilian wheat-growing region, to study inoculation effects of various strains of Azospirillum brasilense and A. amazonense. In all three experiments inoculation with A. brasilense Sp 245 isolated from surface-sterilized wheat roots in Paraná produced the highest plant dry weights and highest N% in plant tops and grain. Grain yield increases with this strain were up to 31 % but were not significant. The application of 60 or 100 kg N ha^–1 to the controls increased N accumulation and produced yields less than inoculation with this strain. Another A. brasilense strain from surface-sterilized wheat roots (Sp 107st) also produced increased N assimilation at the lower N fertilizer level but reduced dry weights at the high N level, while strain Sp 7 + Cd reduced dry weights and N% in the straw at both N levels. The A. amazonense strain isolated from washed roots and a nitrate reductase negative mutant of strain Sp 245 were ineffective. Strains Sp 245 and Sp 107st showed the best establishment within roots while strain Cd established only in the soil.     

Seedlings growth promotion by Azospirillum brasilense under normal and drought conditions remains unaltered in Tebuconazole-treated wheat seeds

The wide use of pesticides in modern agriculture may cause side effects on the non-target microflora. Data on the fungicide Tebuconazole effects on Azospirillum-wheat association are scarce. We analyzed the effects of Tebuconazole on: (a) Azospirillum brasilense Sp245 growth in pure culture, (b) A. brasilense Sp245 colonization of Triticum aestivum cv ProINTA Oasis roots, (c) A. brasilense Sp245-inoculated seedlings growth under normal and water stress conditions in the presence of 20% polyethylene glycol 8000. Seeds were separated in Tebuconazole-free and Tebuconazole-treated lots. Inoculated and non-inoculated seedlings were grown in hydroponics in the dark at 20 °C for 72 h. Root surface, coleoptile length, fresh and dry (DW) weights in both tissues and diazotrophic bacterial most probable number in roots were determined. Water contents and shoot-to-roots DW ratio were calculated. Neither Azospirillum growth nor root colonization was affected by Tebuconazole. Under normal growth conditions most of the growth parameters analyzed, revealed a clear positive effect of A. brasilense on wheat seedlings up to 72 h treatments. The characteristic Azospirillum enhancing effects observed on roots remained unaltered by Tebuconazole. The present study shows that Tebuconazole is compatible with A. brasilense Sp245-wheat inoculation.     

Host-plant specificity in the infection of cereals with Azospirillum spp

The specificity of the infection of maize, wheat and rice roots by N₂-fixing Azospirillum spp was studied in four greenhouse experiments using pots with unsterilized soil and in two field experiments. In all experiments A. lipoferum was most frequently isolated from externally sterilized roots of maize, and A. brasilense nir^? (nitrite reductase negative) from wheat and rice. In pot experiments, A. brasilense nir⁺ was isolated with moderate frequency from within maize roots but rarely from within wheat or rice roots. Inoculation of the pots with a mixture of representative strains of the three Azospirillum groups had no effect on the proportion of strains recovered from each plant species. In the field experiments, inoculation with spontaneous streptomycin-resistant mutants of two of the representative strains confirmed the apparent specificity of A. lipoferum for maize roots and of A. brasilense for wheat but the results were partially obscured by the unexpectedly high proportion of streptomycin-resistant strains isolated from within the roots of uninoculated plants.     

Effect of Azospirillum brasilense inoculation on rhizobacterial communities analyzed by denaturing gradient gel electrophoresis and automated ribosomal intergenic spacer analysis

Nucleic acid-based techniques allow the exploration of microbial communities in the environments such as the rhizosphere. Azospirillumbrasilense, a plant growth promoting rhizobacterium (PGPR), causes morphological changes in the plant root system. These changes in root physiology may indirectly affect the microbial diversity of the rhizosphere. In this study, the changes in the rhizobacterial structure following A. brasilense inoculation of maize (Zea mays) plants was examined by PCR-denaturating gradient gel electrophoresis (DGGE) and automated ribosomal intergenic spacer analysis (ARISA), using two universal primers sets for the 16S rRNA gene, and an intergenic 16S-23S rDNA primer set, respectively. Similar results were obtained when using either ARISA or DGGE performed with these different primer sets, and analyzed by different statistical methods: no prominent effect of A. brasilense inoculation was observed on the bacterial communities of plant roots grown in two different soils and in different growth systems. In contrast, plant age caused significant shifts in the bacterial populations.     

Phenotypic variation in Azospirillum brasilense Sp7 does not influence plant growth promotion effects

The Azospirillum genus comprises free-living, plant growth-promoting, nitrogen-fixing bacteria found in the rhizosphere of plant roots. Azospirilla are able to promote plant growth mainly through improvement of root development. Bacterial surface components, such as extracellular polysaccharides and proteins, are involved in root colonization. Phase variation – or phenotypic variation – is one of the mechanisms by which microorganisms adapt to environmental changes. This phenomenon is characterized by the presence of a sub-population of the bacteria presenting a different phenotype relative to the major population. In this study we characterized phenotypic variation of Azospirillum brasilense Sp7. When plated on solid media, some A. brasilense colonies were shown to possess a much more mucoid morphology, producing 7.5–8 times more exopolysaccharide with different monosaccharide composition than the parental strain Sp7. The rate of appearance of this kind of variant colonies was 1 in 5000, in agreement with the accepted rate for the phase/phenotypic variation phenomenon. The variants were significantly more resistant to heat and UV-exposure than the parental strain and displayed genomic changes as seen by the different band patterns following ERIC-PCR, BOX-PCR and RAPD analyses. In plant inoculation experiments under greenhouse conditions, with maize, wheat, soybean and peanuts, the EPS overproducing variants performed as similar as the parental strain. Therefore, EPS overproduction did not confer an apparent advantage to A. brasilense in terms of induction of plant growth promotion.     

Suppression of damping-off of cucumber caused by Pythium ultimum with live cells and extracts of Serratia marcescens N4-5

Environmentally friendly control measures are needed for the soil-borne pathogen, Pythium ultimum. This pathogen can cause severe losses to field- and greenhouse-grown cucumber and other cucurbits. Live cells and ethanol extracts of cultures of the bacterium Serratia marcescens N4-5 provided significant suppression of damping-off of cucumber caused by P. ultimum when applied as a seed treatment. Live cells of this bacterium also suppressed damping-off caused by P. ultimum on cantaloupe, muskmelon, and pumpkin. Culture filtrates from strain N4-5 contained chitinase and protease activities while ethanol extracts contained the antibiotic prodigiosin, the surfactant serrawettin W1, and possibly other unidentified surfactants. Production of prodigiosin and serrawettin W1 was temperature-dependent, both compounds being detected in extracts from N4-5 grown at 28 °C but not in extracts from N4-5 grown at 37 °C. Ethanol extracts from strain N4-5 grown at 28 °C inhibited germination of sporangia and mycelial growth by P. ultimum in in vitro experiments. There was no in vitro inhibition of P. ultimum associated with ethanol extracts of strain N4-5 grown at 37 °C. Prodigiosin, purified from two consecutive thin-layer chromatography runs using different solvent systems, inhibited germination of sporangia and mycelial growth of P. ultimum. Another unidentified compound(s) also inhibited germination of sporangia but did not inhibit mycelial growth. There was no in vitro inhibition associated with serrawettin W1. These results demonstrate that live cells and cell-free extracts of S. marcescens N4-5 are effective for suppression of damping-off of cucumber caused by P. ultimum possibly due in part to the production of the antibiotic prodigiosin.     

Peroxidase-antiperoxidase labeling of azospirillum brasilense in field-grown pearl millet

Bacterial cells of Azospirillum brasilense (Tarrand) from their natural habitat were labeled with peroxidase-antiperoxidase (PAP), identified, and observed using the transmission electron microscope. Pure cultures of A. brasilense, axenically inoculated pearl millet root samples, and field-grown inoculated pearl millet root samples were embedded in Luffs araldite. Thin sections were treated using the immunological PAP method. Identification was possible because of the heavy outlining of the cells with a dense deposit of osmium.Pleomorphic forms of A. brasilense were observed in axenic pearl millet root cultures. Encapsulated forms were larger than vibrioid forms, and both types reacted with antiserum against the bacterial strain.     

Potential for enhancement of root growth and nodulation of soybean co-inoculated with Azospirillum and Bradyrhizobium in laboratory systems

The potential enhancement of root growth and nodulation in vegetable soybean (AGS190) was studied with application of Azospirillum brasilense (Sp7) and A. lipoferum (CCM3863) co-inoculated with two Bradyrhizobium japonicum strains (TAL102 and UPMR48). Significant root growth stimulation and nodulation were observed in Azospirillum as well as during its co-inoculation with Bradyrhizobium. Nodule formation is linked with the initiation of new roots; nodules were almost absent even in Bradyrhizobium inoculated plant due to the absence of new roots development in clipped rooted seedlings. Total root length, root number, specific root length, root dry matter, root hair development and shoot dry matter were significantly increased by Azospirillum alone and its co-inoculum. Co-inoculated plants significantly influenced the number of nodules and its fresh weight. A. brasilense seemed to perform better in root growth and nodule development compared to A. lipoferum.     

Rapid identification and discrimination among Egyptian genotypes of Rhizobium leguminosarum bv. viciae and Sinorhizobium meliloti nodulating faba bean (Vicia faba L.) by analysis of nodC, ARDRA, and rDNA sequence analysis

Twenty-eight Rhizobium strains were isolated from the root nodules of faba bean (Vicia faba L.) collected from 11 governorates in Egypt. A majority of these strains (57%) were identified as Rhizobium leguminosarum bv. viciae (Rlv) based on analysis of a nodC gene fragment amplified using specific primers for these faba bean symbionts. The strains were characterized using a polyphasic approach, including nodulation pattern, tolerance to environmental stresses, and genetic diversity based on amplified ribosomal DNA-restriction analysis (ARDRA) of both 16S and 23S rDNA. Analysis of tolerance to environmental stresses revealed that some of these strains can survive in the presence of 1% NaCl and a majority of them survived well at 37 °C. ARDRA indicated that the strains could be divided into six 16S rDNA genotypes and five 23S rDNA genotypes. Sequence analysis of 16S rDNA indicated that 57% were Rlv, two strains were Rhizobium etli, one strain was taxonomically related to Rhizobium rubi, and a group of strains were most closely related to Sinorhizobium meliloti. Results of these studies indicate that genetically diverse rhizobial strains are capable of forming N₂-fixing symbiotic associations with faba bean and PCR done using nodC primers allows for the rapid identification of V. faba symbionts.     

Suppression of root-knot disease by Pseudomonas fluorescens CHA0 in tomato: importance of bacterial secondary metabolite, 2,4-diacetylpholoroglucinol

The antimicrobial metabolites 2,4-diacetylphloroglucinol (2,4-DAPG) and pyoluteorin contribute to the ability of Pseudomonas fluorescens strain CHA0 to control plant diseases caused by soil-borne pathogens. P. fluorescens strain CHA0 and its derivatives CHA89 (antibiotics-deficient) and CHA0/pME3424 (antibiotics overproducing) were investigated as potential biocontrol agents against Meloidogyne javanica the root-knot nematode. Exposure of root-knot nematode to culture filtrates of P. fluorescens under in vitro conditions significantly reduced egg hatch and caused substantial mortality of M. javanica juveniles. Nutrient broth yeast extract (NBY) medium amended with 2% (w/v) glucose or 1 mM EDTA markedly repressed hatch inhibition activity of the strain CHA0 but not that of CHA0/pME3424 or CHA89. On the other hand, NBY medium amended with glucose significantly enhanced nematicidal activity of the strain CHA0/pME3424. Neither glucose nor EDTA had an influence on the nematicidal activity of the strains CHA0 and CHA89. Under in vitro conditions, antibiotic overproducing strain CHA0/pME3424 and CHA0 expressed phl‘-’lacZ reporter gene but strain CHA89 did not. Expression of the reporter gene reflects actual production of DAPG. In general, CHA0/pME3424 expressed reporter gene to a greater extent compared to its wild type counterpart CHA0. Regardless of the bacterial strains, reporter gene expression was markedly enhanced when NBY medium was amended with glucose but EDTA had no such effect. A positive correlation between the degree of juvenile mortality and extent of phl‘-’lacZ reporter gene expression was also observed in vitro. Strain CHA0 produced zones of 4-6 mm on MM medium containing gelatin while strain CHA0/pME3424 and CHA89 did not. When MM medium containing gelatin was amended with 2% glucose of 1 mM EDTA size of haloes produced by the strain CHA0 reduced to 2 mm. Under glasshouse conditions aqueous cell suspension of the strains CHA0 or CHA0/pME3424 at various inoculum levels (10⁷, 10⁸ or 10⁹ cfu ml⁻¹) significantly reduced root-knot development. CHA89 caused significant reduction in galling when applied at 10⁹ cfu ml⁻¹. To better understand the mechanism of nematode suppression, split root bioassay was performed. Split-root experiments, that guarantee a spatial separation of inducing agent and a challenging pathogen, showed that soil treatment of one half of the root system with cell suspension of CHA0 or CHA0/pME3424 resulted in a significant systemic induced resistance leading to reduction of M. javanica infection of tomato roots in the non-baterized nematode treated half. The results clearly suggest that the antibiotic 2,4-DAPG from P. fluorescens CHA0 act as the inducing agents of systemic resistance in tomato roots. Populations of CHA0 and its derivatives declined progressively by 10-fold between first and fourth harvests (0-21 days after inoculation). However, bacterial populations increased at final harvest (28 days after application).     

Plant growth promoting properties of a strain of Enterobacter ludwigii isolated from Lolium perenne rhizosphere

The capability of native bacterial strains isolated from Lolium perenne rhizosphere to behave as plant growth promoting bacteria and /or biocontrol agents was investigated. One strain (BNM 0357) over 13 isolates from the root tips of L. perenne resulted proved to be nitrogenase positive (ARA test) and an IAA producer. Conventional tests and the API 20E diagnostic kit indicated that BNM 0357 behaves to the Enterobacteriaceae family and to the Enterobacter genus. Molecular identification by 16S rRNA sequence analysis indicated that BNM 0357 had the highest similarity to Enterobacter ludwigii (EN-119). Isolate BNM 0357 had the capability to solubilize calcium triphosphate and to antagonize Fusarium solani mycelial growth and spore germination. Strain BNM 0357 also showed the ability to improve the development of the root system of L. perenne. This study disclosed features of E. ludwigii BNM 0357 that deserve further studies aimed at confirming its putative importance as a PGPR.     

Genetic diversity among Elaeagnus compatible Frankia strains and sympatric-related nitrogen-fixing actinobacteria revealed by nifH sequence analysis

Elaeagnus compatible Frankia isolates from Tunisian soil have been previously clustered with Frankia, colonizing Elaeagnaceae and Rhamnaceae in two different phylogenetic subgroups, while strain BMG5.6 was described as a new lineage closely related to Frankia and Micromonospora genera. In this study we further assess the diversity of captured Frankia and the relationship with BMG5.6-like actinobacteria, by using nifH gene sequences. Using PCR-RFLP screening on DNA extracted from lobe nodules, additional microsymbionts sharing BMG5.6 features have been detected proving a widespread occurrence of these actinobacteria in Elaeagnus root nodules. Neighbour-Joining trees of Frankia nifH sequences were consistent with previously published 16S rRNA and GlnII phylogenetic trees. Although four main clades could be discerned, actinobacterial strain BMG5.6 was clustered with Frankia strains isolated from Elaeagnus. The present study underscored the emanation of new diazotrophic taxon isolated from actinorhizal nodules occupying intermediate taxonomic position between Frankia and Micromonospora. Moreover, its aberrant position in nifH phylogeny should open network investigations on the natural history of nitrogen-fixing gene among actinobacteria.     

Assessment of the plant growth promotion abilities of six bacterial isolates using Zea mays as indicator plant

Zea mays, one of the most important cereals worldwide, is a plant not only with food and energy value, but also with phytoremediation potential. The use of plant growth promoting (PGP) rhizobacteria may constitute a biological alternative to increase crop yield and plant resistance to degraded environments. In search for PGP rhizobacteria strains, 6 bacterial isolates were isolated from a metal contaminated site, screened in vitro for their PGP characteristics and their effects on the growth of Z. mays were assessed. Isolates were identified as 3A10^T, ECP37^T, corresponding to Chryseobacterium palustre and Chryseobacterium humi, and 1ZP4, EC15, EC30 and 1C2, corresponding to strains within the genera Sphingobacterium, Bacillus, Achromobacter, and Ralstonia, respectively. All the bacterial isolates were shown to produce indole acetic acid, hydrogen cyanide and ammonia when tested in vitro for their plant growth promoting abilities, but only isolates 1C2, 1ZP4 and ECP37^T have shown siderophore production. Their further application in a greenhouse experiment using Z. mays indicated that plant traits such as root and shoot elongation and biomass production, and nutrient status, namely N and P levels, were influenced by the inoculation, with plants inoculated with 1C2 generally outperforming the other treatments. Two other bacterial isolates, 1ZP4 and ECP37^T also led to increased plant growth in the greenhouse. These 3 species, corresponding to strains within the genera Ralstonia (1C2), Sphingobacterium (1ZP4), and to a strain identified as C. humi (ECP37^T) can thus be potential agents to increase crop yield in maize plants.     

Cadmium-tolerant plant growth-promoting bacteria associated with the roots of Indian mustard (Brassica juncea L. Czern.)

Eleven cadmium-tolerant bacterial strains were isolated from the root zone of Indian mustard (Brassica juncea L. Czern.) seedlings grown in Cd-supplemented soils as well as sewage sludge and mining waste highly contaminated with Cd. The bacteria also showed increased tolerance to other metals including Zn, Cu, Ni and Co. The isolated strains included Variovorax paradoxus, Rhodococcus sp. and Flavobacterium sp., and were capable of stimulating root elongation of B. juncea seedlings either in the presence or absence of toxic Cd concentrations. Some of the strains produced indoles or siderophores, but none possessed C₂H₂-reduction activity. All the strains, except Flavobacterium sp. strain 5P-3, contained the enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase, which hydrolyses ACC (the immediate precursor of plant hormone ethylene) to NH₃ and α-ketobutyrate. V. paradoxus utilized ACC as a sole source of N or energy. A positive correlation between the in vitro ACC deaminase activity of the bacteria and their stimulating effect on root elongation suggested that utilization of ACC is an important bacterial trait determining root growth promotion. The isolated bacteria offer promise as inoculants to improve growth of the metal accumulating plant B. juncea in the presence of toxic Cd concentrations and for the development of plant-inoculant systems useful for phytoremediation of polluted soils.     

Size, symbiotic effectiveness and genetic diversity of field pea rhizobia (Rhizobium leguminosarum bv. viciae) populations in South Australian soils

Field pea (Pisum sativum L.) is widely grown in South Australia (SA), often without inoculation with commercial rhizobia. To establish if symbiotic factors are limiting the growth of field pea we examined the size, symbiotic effectiveness and diversity of populations of field pea rhizobia (Rhizobium leguminosarum bv. viciae) that have become naturalised in South Australian soils and nodulate many pea crops. Most probable number plant infection tests on 33 soils showed that R. l. bv. viciae populations ranged from undetectable (six soils) to 32×10³ rhizobia g⁻¹ of dry soil. Twenty-four of the 33 soils contained more than 100 rhizobia g⁻¹ soil. Three of the six soils in which no R. l. bv. viciae were detected had not grown a host legume (field pea, faba bean, vetch or lentil). For soils that had grown a host legume, there was no correlation between the size of R. l. bv. viciae populations and either the time since a host legume had been grown or any measured soil factor (pH, inorganic N and organic C). In glasshouse experiments, inoculation of the field pea cultivar Parafield with the commercial Rhizobium strain SU303 resulted in a highly effective symbiosis. The SU303 treatment produced as much shoot dry weight as the mineral N treatment and more than 2.9 times the shoot dry weight of the uninoculated treatment. Twenty-two of the 33 naturalised populations of rhizobia (applied to pea plants as soil suspensions) produced prompt and abundant nodulation. These symbioses were generally effective at N₂ fixation, with shoot dry weight ranging from 98% (soil 21) down to 61% (soil 30) of the SU303 treatment, the least effective population of rhizobia still producing nearly double the growth of the uninoculated treatment. Low shoot dry weights resulting from most of the remaining soil treatments were associated with delayed or erratic nodulation caused by low numbers of rhizobia. Random amplified polymorphic DNA (RAPD) polymerase chain reaction (PCR) fingerprinting of 70 rhizobial isolates recovered from five of the 33 soils (14 isolates from each soil) showed that naturalised populations were composed of multiple (5-9) strain types. There was little evidence of strain dominance, with a single strain type occupying more than 30% of trap host nodules in only two of the five populations. Cluster analysis of RAPD PCR banding patterns showed that strain types in naturalised populations were not closely related to the current commercial inoculant strain for field pea (SU303, ≥75% dissimilarity), six previous field pea inoculant strains (≥55% dissimilarity) or a former commercial inoculant strain for faba bean (WSM1274, ≥66% dissimilarity). Two of the most closely related strain types (≤15% dissimilarity) were found at widely separate locations in SA and may have potential as commercial inoculant strains. Given the size and diversity of the naturalised pea rhizobia populations in SA soils and their relative effectiveness, it is unlikely that inoculation with a commercial strain of rhizobia will improve N₂ fixation in field pea crops, unless the number of rhizobia in the soil is very low or absent (e.g. where a legume host has not been previously grown and for three soils from western Eyre Peninsula). The general effectiveness of the pea rhizobia populations also indicates that reduced N₂ fixation is unlikely to be the major cause of the declining field pea yields observed in recent times.     

Azospirillum brasilense mitigates water stress imposed by a vascular disease by increasing xylem vessel area and stem hydraulic conductivity in tomato

Water stress, with its negative consequences on plant growth and survival, can be mitigated by Azospirillum brasilense inoculation. In tomato, A. brasilense delays wilting caused by a vascular pathogen, Clavibacter michiganensis subsp. michiganensis, by yet unknown mechanisms. We studied morphological, anatomical and physiological changes induced by A. brasilense in tomato that relate to water stress tolerance, which could explain the deferral in symptom expression. For this purpose, tomato seeds were treated or not with A. brasilense BNM65, and 5 weeks later plants were challenged with C. michiganensis subsp. michiganensis or mock inoculated with water. There was a large growth promotion associated to Azospirillum: treated plants had higher total biomass and leaf area. In relation to water stress tolerance, Azospirillum treated plants had larger xylem vessel area, higher stem specific hydraulic conductivity, thicker stems, and lower shoot/root dry matter and specific leaf area. These changes were opposite to those induced by C. michiganensis subsp. michiganensis. We conclude that A. brasilense favoured a better adjustment of plant-water relations by several mechanisms, and thus, transitorily alleviated symptoms expression of a vascular disease.     

甘薯长喙壳侵染对甘薯块根抗氧化酶活性的影响

以易感黑斑病甘薯品种“徐薯18”为试验材料, 分别接种分离自石榴、甘薯和芋头的甘薯长喙壳(Ceratocystis fimbriata), 研究其保护酶活性及丙二醛(MDA)含量变化。结果显示, 非亲和性菌株[分离自石榴的甘薯长喙壳(CQS2)和分离自芋头的甘薯长喙壳(YP1)]和亲和性菌株[分离自甘薯的甘薯长喙壳(SP1)]接种的甘薯块根苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性, 与对照处理相比均有升高, 且接种CQS2 和YP1 的处理始终高于接种SP1 的处理; 非亲和性菌株和亲和性菌株接种的甘薯块根MDA 含量均呈先上升后下降趋势, 且接种CQS2 和YP1 的处理在接种25 h 时MDA 含量明显高于接种SP1 的处理, 接种SP1 的处理在接种后期(50 h)的MDA 含量降至与对照相当水平。由此表明: 不同寄主的甘薯长喙壳侵染甘薯块根后, 非亲和性接种菌株(CQS2 和YP1)比亲和性接种菌株(SP1)能增强POD、SOD和CAT 酶活性及诱导较多的MDA, 从而增强病原菌侵染的抗性。