Control of Fusarium wilt in carnation grown on rockwool by Pseudomonas sp. strain WCS417r and by Fe-EDDHA

In carnations grown on rockwool disease incidence of fusarium wilt caused byFusarium oxysporum f.sp.dianthi (Fod) was reduced when Fe-EDDHA instead of Fe-DTPA was used as iron source in the nutrient solution. Addition ofPseudomonas sp. strain WSC417r intensified this reduction in the cultivar Pallas, moderately resistant to Fusarium, but not in the susceptible cultivar Lena. Treatment of plants with Fe-EDDHA instead of Fe-DTPA as iron source resulted in higher numbers and percentages on the roots, ofin vitro antagonistic fluorescent pseudomonads. However, differences were only significant at 56 days after planting for cv. Lena and at 14 and 28 days after planting for cv. Palas. Both chelators, at different concentrations, had no effect on root colonization by eitherPseudomonas sp. strain WCS417r orFod strain WCS816. However, when coinoculated, reduced numbers of propagules ofFusarium were found at concentrations of Fe-EDDHA lower than 10^–5 M.Higher concentrations of the siderophore fusarine produced byFod strain WCS816 were demonstrated when Fe-EDDHA instead of Fe-DTPA was used as iron source in culture media. At equal concentrations, no such differences were found in the amount of siderophore produced by WCS417r. Germ tube length ofFod was less with Fe-EDDHA than with Fe-DTPA. The reduction of germ tube length was stronger when the purified siderophore of WCS417r was added in excess to the culture media with Fe-EDDHA than those with Fe-DTPA. Therefore, the observed reduction of germ tube growth can not completely be explained by iron deprivation. It appeared that EDDHA exhibited a toxic effect for conidia ofFod strain WCS816 as well.we conclude that the observed disease reduction by Fe-EDDHA is a consequence of a limitation of iron availability forFod. This limitation is possibly intensified by the increase in number or percentage of antagonistic fluorescent pseudomonads that strongly compete for iron. The additional effect after bacterization withPseudomonas strain WCS417r in Fe-EDDHA treated carnations of cv. Pallas is likely to be due, at least partly, to a direct competition for iron between the siderophores ofFod strain WCS816 and ofPesudomonas sp. strain WCS417r.Samenvatting Verwelkingsziekte in anjers op steenwol, veroorzaakt doorFusarium oxysporum f. sp.dianthi (Fod), werd gereduceerd indien het ijzer-chelaat Fe-EDDHA in plaats van Fe-DTPA werd toegevoegd aan de nutriëntenvloeistof. Bacterisatie metPseudomonas sp. stam WCS417r had een additioneel effect bij de matig resistence cultivar Pallas maar niet bij de vatbare cultivar Lena. Toevoeging van Fe-EDDHA in plaats van Fe-DTPA aan planten als ijzerbron resulteerde op de wortels in hogere aantallen en percentages fluorescerende pseudomonaden, diein vitro antagonistisch waren ten opzichte vanFod. De verschillen waren echter alleen significant 56 dagen na planten voor de cultivar Lena en 14 en 28 dagen na planten voor de cultivar Pallas. Beide chelaten vertoonden bij verschillende concentraties geen effect op de kolonisatie van de wortel door beide microorganismen. Echter, wanneer beide micro-organismen gezamelijk werden toegevoegd nam de wortelkolonisatie doorFod stam WCS816 af bij concentraties lager dan 10^–5 M Fe-EDDHA. Er werd meer van het siderofoor fusarine doorFod stam WCS816 geproduceerd bij concentraties lager dan 10^–4 M Fe indien Fe-EDDHA in plaats van Fe-DTPA als ijzerbron aan het cultuurmedium was toegevoegd. Er werd geen effect van beide chelaten gevonden op de siderofoorproduktie door WCS417r. Indien een overmaat van het gezuiverde siderofoor van WCS417r werd toegevoegd aan Fe-EDDHA werden een sterkere afname van de kiembuislengte gevonden dan toevoeging aan Fe-DTPA. De reductie van de kiembuislengte bleek niet volledig verklaard te kunnen worden door een afname van de ijzerbeschikbaarheid. Het chelaat EDDHA heeft ook een toxisch effect op conidiën van fusarium.Wij concluderen, dat de waargenomen reductie van de verwelkingziekte door Fe-EDDHA een gevolg is van de afname van de ijzerbeschikbaarheid voorFod. Dit wordt waarschijnlijk versterkt door de ontwikkeling van een antagonistische, fluorescerendePseudomonas-populatie die sterk concurreren om ijzer. Het additioneel effect dat door bacterisatie metPseudomonas sp. WCS417r van de met Fe-EDDHA behandelde matig resistante anjers (Pallas) werd verkregen is voor een deel het gevolg van een directe concurrentie om ijzer tussen de sideroforen vanFod stam WCS816 en vanPseudomonas sp. stam WCS417r.     

Potato diseases caused by soft rot erwinias: an overview of pathogenesis

Three soft rot erwinias, Erwinia carotovora ssp. carotovora , E. carotovora ssp. atroseptica and E. chrysanthemi are associated with potatoes causing tuber soft rot and blackleg (stem rot). Latent infection of tubers and stems is widespread. As opportunistic pathogens, the bacteria tend to cause disease when potato resistance is impaired. Pathogenesis or disease development in potato tubers and stems is discussed in terms of the interaction between pathogen, host and environment, microbial competition and recent findings on the molecular basis of pathogenicity. Emphasis is placed on the role of free water and anaerobiosis in weakening tuber resistance and in providing nutrient for erwinias to multiply. Blackleg symptoms are expressed when erwinias predominate in rotting mother tubers, invade the stems and multiply in xylem vessels under favourable weather conditions. Soft rot erwinias tend to out-compete other bacteria in tuber rots because of their ability to produce larger quantities of a wider range of cell wall-degrading enzymes. However, despite extensive studies on their induction, regulation and secretion, little is known about the precise role of the different enzymes in pathogenesis. The putative role of quorum-sensing regulation of these enzymes in disease development is evaluated. The role certain pathogenicity-related characters, including motility, adhesion, siderophores, detoxifying systems and the hrp gene complex, common to most bacteria including symbionts and saprophytes, could play in latent and active infections is also discussed.     

Modulation of Uptake and Translocation of Iron and Copper from Root to Shoot in Common Bean by Siderophore-Producing Microorganisms

Microbes have developed high-affinity uptake mechanisms to assimilate iron (Fe) and other metals such as aluminum (Al), gallium (Ga), chromium (Cr), and copper (Cu). Siderophores, which are metal chelating compounds, and membrane receptor proteins are involved in these specialized mechanisms. A few siderophore-producing microorganisms associated with plant roots also influence the uptake of some metals. In this study, the potential microbial-assisted Cu and Fe uptake by Phaseolus vulgaris (common bean) plants was evaluated. Seedlings of cultivated common bean varieties Bayo-INIFAP (B) and Negro-150 (N) and wild types yellowish (WY) and black (WB) were developed in the presence of a Cu and Fe solution and associated with the siderophore-producing microorganisms R. leguminosarumbv. Phaseoli (strains 19, 44, and 46); Pseudomonas fluorescens(strain Avm), and Azospirillum brasilense (strain 154). Seedlings of cultivated variety N and black wild type WB inoculated with the strain CPMex.44 accumulated 71% and 30% more Fe than the un-inoculated plants, respectively; however, the wild black bean accumulated the highest absolute amount of Fe (221.56 mg/kg of dry matter) as compared with the cultivated black variety N (126.16 mg/kg of dry matter) (P < 0.05). In the wild type WY seedlings, the highest Fe accumulation was observed when the seeds were inoculated with the Pseudomonas strain Avm (206 mg/kg of dry matter) (P < 0.05). The interaction of Pseudomonas strain Avm with seedlings of the cultivated B variety and the wild type WB promoted the highest accumulation of Cu (51 and 54 mg/kg of dry matter, respectively), 7 and 14 mg more than in the respective non-inoculated seedlings. No promotion of Fe accumulation was observed in the seedlings of the cultivated B variety and in roots; instead, less Fe was accumulated. The wild type WY did not show any improvement in Cu accumulation. In this study, Rhizobiumstrains promoted Fe but not Cu uptake in P. vulgaris seedlings while Pseudomonas strains promoted the uptake of both Cu and Fe.     

Mobilization of glucose‐6‐phosphate from ferrihydrite by ligand‐promoted dissolution is higher than of orthophosphate

We investigated glucose‐6‐phosphate (G6P) release from ferrihydrite by ligand‐promoted dissolution by oxalate, ascorbate, and desferrioxamine‐B in comparison to orthophosphate (OP). Overall, P release was poorly related to Fe dissolution. Initial release of G6P was higher as compared to OP, likely due to its preferential adsorption at outer surface sites caused by pore size effects. Our data suggest that G6P is potentially more bioavailable (i.e ., mobilizable) than OP.     

A novel iron transporter in Streptococcus iniae

Streptococcus iniae is a major pathogen that results in considerable economic loss to fish farms. Restricted availability of iron is a huge obstacle to survival for pathogenic bacteria during infection, and iron acquisition is important in bacterial virulence. In this study, S. iniae HD‐1 was shown not to produce siderophores (low‐molecular‐weight compounds) but rather to require iron‐containing proteins for growth under iron‐restricted conditions. The adenosine triphosphate (ATP)‐binding‐cassette (ABC) transporter system (ftsABCD), which is cotranscribed by four downstream genes, namely, ftsA, ftsB, ftsC and ftsD, was identified as responsible for haem utilization of S. iniae. Analysis of the corresponding recombinant protein, FtsB, indicated that it is a putative lipoprotein which plays a role in haem utilization and is produced in vivo during infection with S. iniae HD‐1, and therefore may be a potential candidate antigen for a streptococcal vaccine.     

Metal-induced oxidative stress impacting plant growth in contaminated soil is alleviated by microbial siderophores

High levels of metals impede plant growth by affecting physiological processes. Siderophores are microbial Fe-chelators that, however, bind other metals. This study evaluated plant growth in a soil containing elevated levels of metals, including Al, Cu, Fe, Mn, Ni, and U, using Streptomyces-derived cell-free supernatant containing siderophores and auxins. Cowpea plants in the soil were treated with the culture filtrate. Growth was measured and biochemical analyses such as chlorophyll contents, RNA and protein quantification, lipid membrane peroxidation, and anti-oxidative responses were conducted to evaluate oxidative stress in the plants. Liquid chromatography-mass spectrometry was used to simulate competition for siderophore binding, and metal content of plants was determined spectroscopically. Whereas the metals inhibited plant growth, addition of siderophores improved growth. There was evidence of lipid peroxidation, an enhanced superoxide dismutase activity, and lowered chlorophyll, RNA, protein, carotenoid and residual indole acetic acid contents, especially in control plants. Siderophore competition assays between Al and Fe, and Fe and Cu suggested that trivalent metals are more competitive for siderophore binding than divalent ones. Compared to control plants, higher amounts of metals were obtained in siderophore-treated plants. Siderophores were able to supply plants with Fe in the presence of levels of metals, mainly Al, Cu, Mn, Ni and U that otherwise inhibit Fe acquisition. This led to enhanced chlorophyll content, circumventing lipid peroxidation effects on leaves. Siderophores lowered the formation of free radicals, thereby protecting microbial auxins from degradation and enabling them to enhance plant growth which in turn resulted in augmented metal uptake.     

Density,metabolic activity,and identity of cultivable rhizosphere bacteria on Salix viminalis in disturbed arable and landfill soils

The objective of this study was to provide fundamental data for a subsequent selection of willow growth and soil remediation promoting bacterial strains. The rhizosphere of willows (Salix viminalis) was screened for cultivable bacteria with high enzymatic activity (proteolytic, pectolytic, cellulolytic, amylolytic) and production of siderophores at four test sites with broad spectrum of anthropogenic soil disturbance: sewage‐sludge application, impoverishment by unfavorable arable use, ash dumping, and household‐waste depositing. The density of bacteria in the rhizosphere ranged from 7.92 to 8.56 log₁₀ of colony‐forming units per gram dry weight of soil and varied in a site‐ and willow‐clone‐specific manner. Within the 240 bacterial strains, a high diversity of metabolic activities was observed but was rarely combined in one strain (1.2% having six and 5.8% having five out of seven metabolic activities, respectively). The majority of strains (79.2%) revealed just one or two metabolic activities. Most common was a combination of lipolytic, proteolytic activities, and siderophore production as found in 13.8% of the bacterial strains. The 50 strains with the highest metabolic activity belonged predominantly to the Gammaproteobacteria (66%), the others to Flavobacteria (18%), Betaproteobacteria (8%), Actinobacteria (4%), and Bacilli (4%). The highest portion of cultivable strains of rhizosphere bacteria with high metabolic activities belonged to the genera Pseudomonas, Serratia, and Flavobacterium. We hypothesize that these genera include strains that support willow growth and soil remediation. Therefore, the described strain collection from the rhizosphere of S. viminalis provides a valuable basis for a subsequent selection of these candidates for applications in improvement of site adaptation of plants or remediation of soils.     

荧光假单胞杆菌的嗜铁素是控制桉树灰霉病的主要因子

 本文对3个假单胞杆菌菌株(Pseudomonas spp.)及其嗜铁素(pseudobactin siderophore)缺失突变体防治桉树灰霉病进行了研究.平板拮抗活性测定表明,荧光假单胞杆菌(P.fluorescens) WCS374r菌株和恶臭假单胞杆菌(P.putida) WCS358r菌株通过对铁离子的竞争抑制灰霉菌的生长.在接种灰霉病菌之前10 h将WCS358r、WCS374r和WCS417r施用于受伤的桉树叶片后,可分别降低发病率48.9%、58.3%和40.3%;当将3种生防菌分别与灰霉病菌混合后接种桉树叶片,WCS358r和WCS374r仍然能够显著地降低发病率;在接种灰霉病菌12 h后再施用生防菌,WCS358r和WCS374r对病菌仍具有一定的抑制作用,而在24 h后施用生防菌,3个菌株均未表现显著的防治效果.WCS358r和WCS417r的嗜铁素缺失突变体无防病作用,而WCS374r的嗜铁素缺失突变体虽然还能有效地防治灰霉病,但与WCS374r相比,防病效果减弱.本试验结果说明假单胞杆菌的嗜铁素是控制桉树灰霉病的重要因子.     

Delftia sp.B9对镉胁迫下水稻种子萌发及幼苗镉积累的影响

为探讨耐镉细菌Delftia sp. B9对镉(Cd)胁迫下水稻种子萌发及幼苗吸收积累Cd的影响,以两种水稻(华润2号、深两优5814)为材料,研究水稻在3种Cd胁迫浓度(0、0.01、0.1 mg·L~(-1))下添加Delftia sp. B9菌液对水稻幼苗生长和积累Cd的影响。结果表明:Cd胁迫浓度为0.1 mg·L~(-1)时,Delftia sp. B9产吲哚乙酸(IAA)能力与对照相比显著减少2.87 mg·L~(-1),产铁载体相对含量下降17.34%。Cd胁迫浓度为0.1 mg·L~(-1)时,添加Delftia sp. B9菌液对水稻种子萌发和耐性系数有显著的促进作用。Cd胁迫下添加Delftia sp. B9菌液的处理(T3)与对照(T1)相比能显著增加两种水稻幼苗的根长、株高、叶绿素a和叶绿素b含量。添加Delftia sp.B9显著降低两种水稻幼苗根、茎、叶中Cd含量,使华润2号根、茎、叶中Cd含量分别降低63.81%、67.59%、70.84%,使深两优5814根、茎、叶中Cd含量分别降低75.95%、74.84%、80.81%。研究表明,耐镉细菌Delftia sp. B9可促进Cd胁迫下水稻种子萌发,增加水稻幼苗叶绿素含量和株高,并降低根、茎、叶中Cd含量。