Factors Influencing Nodule Occupancy by Inoculant Rhizobia

Inoculation of legumes under field conditions with superior nitrogen-fixing rhizobia does not always result in the desired yield increase. Often it is observed that the inoculum strain fails to occupy a significant proportion of the nodules. The introduced inoculant strains have to compete with the indigenous, often ineffective, nitrogen-fixing rhizobial population at different levels. The success of inoculation depends to a large extent on the ratio of the inoculant cells to indigenous rhizobia. However, intrinsic characteristics of the inoculant and indigenous rhizobia, and their responses to abiotic and biotic environmental variables, also influence the outcome of inoculation. In this review, the genetic basis for “efficient host-bacteria interaction” is reviewed. In addition, environmental factors that influence competition and saprophytic competence of rhizobia are discussed.     

Water and Seed Survival

Between about –350 and –14 MPa the rate of lossof viability in orthodox seeds is a positive function of waterpotential. The relative effect of water potential has been analysedin an oily seed (lettuce) and a non-oily seed (barley) and foundto be more or less identical. The lower limit for the relationin various species coincides with a seed moisture content (wetbasis) between about 2 and 6%. Below this level there is littleor no improvement in longevity with reduction in moisture content.The upper limit coincides with moisture contents of between15 and 28%, depending on whether the seeds are oily or non-oily.A water potential of about –14 MPa is the threshold forrespiration which increases more-or-less linearly with increasein water potential above this level. Above this threshold, andproviding oxygen is available to sustain respiration, seed longevityincreases with increase in water potential except that, unlessthe seeds are dormant, germination may be initiated at a waterpotential of about –1·5 to –0·5 MPa.In the absence of oxygen there may be a slight further declinein longevity with increase in water potential above –14MPa before longevity reaches a minimum value Since they cannot be dried very much without immediate lossof viability, recalcitrant seeds survive longest in the presenceof oxygen at maximum water potential commensurate with preventinggermination. The threshold water potential for immediate lossof viability has not been determined for most species but itis probable that it is close to the water potential typicalof the permanent wilting point in these plants, say –2MPa Lactuca saliva L., lettuce, Hordeum oulgare L., barley, seed storage, moisture content, relative humidity, water potential, temperature, oxygen     

Invasion Pressure to a Ballast-flooded Estuary and an Assessment of Inoculant Survival

The relationships between invasion pressure, post-transport inoculant survival, and regional susceptibility to invasion are poorly understood. In marine ecosystems, the movement and release of ballast water from ocean-going ships provides a model system by which to examine the interplay among these factors. One of the largest estuaries in North America, the Chesapeake Bay, receives tremendous amounts of foreign ballast water annually and thus should be at high invasion risk. To date, however, few introductions in Chesapeake Bay have been attributed to ballast release. To understand better the dynamics of this invasion process, we (1) characterized and quantified the biota arriving to Chesapeake Bay in foreign ballast water, (2) compared temperatures and salinities of ballast water and harbor water in upper Chesapeake Bay, and (3) tested experimentally survival of organisms collected from ballast water in temperatures and salinities characteristic of the region. From 1993 to 1994, we sampled planktonic and benthic organisms from 60 foreign vessels arriving to Chesapeake Bay. Our data show that the estuary is being inoculated by a diverse assemblage of aquatic organisms from around the world. Furthermore, the short transit time (15d) for most vessels ensured that substantial numbers of larval and post-larval organisms were being deballasted alive. Most of the ballast water discharged into the upper Chesapeake Bay, however, was significantly higher in salinity (>20) than that of the receiving harbor. In laboratory tolerance experiments, ballast water organisms perished under such conditions. Thus, a mismatch in physical conditions between donor and receiver regions may explain the dearth of invasions in the upper Bay. It is likely that the lower Chesapeake Bay, which is more saline, remains at higher risk to ballast water invasion. Recognition of such intraregional differences should allow more focused predictions for monitoring and management.     

Residual Impact of the Biocontrol Inoculant Pseudomonas fluorescens F113 on the Resident Population of Rhizobia Nodulating a Red Clover Rotation Crop

A field trial was previously conducted in which sugarbeet seeds were either untreated, inoculated with the biocontrol strain Pseudomonas fluorescens F113Rif, or treated with chemical fungicides. Following harvest of sugarbeet, the field site was sown with uninoculated red clover. The aim of this study was to assess the residual impact of the microbial inoculant (and the fungicide treatment) on the diversity of resident rhizobia nodulating the red clover rotation crop. The percentage of nodules yielding rhizobial isolates after surface disinfection was 67% in the control and 70% in the P. fluorescens F113Rif treatment, but only 23% in the chemical treatment. Isolates were characterized by RAPD analysis. The main RAPD cluster (arbitrarily defined at 70% similarity) was prevalent in all three treatments. In addition, the distribution of RAPD clusters followed a log series model, regardless of the treatment applied, indicating that neither the microbial inoculant nor the fungicide treatment had caused a strong perturbation of the rhizobial population. When the P. fluorescens F113Rif and control treatments were compared using diversity indices, however, it appeared that the genetic diversity of rhizobia was significantly less in the inoculated treatment. The percentage of rhizobia sensitive to 2,4-diacetylphloroglucinol (Phl; the antimicrobial metabolite produced by P. fluorescens F113Rif) fluctuated according to field site heterogeneity, and treatments had no effect on this percentage. Yet, the proportion of Phl-sensitive isolates in the main RAPD cluster was lower in the P. fluorescens F113Rif treatment compared with the control, raising the possibility that the residual impact of the inoculant could have been partly mediated by production of Phl. This impact on the rhizobial population took place without affecting the functioning of the Rhizobium–clover symbiosis.     

Survival of Cowpea Rhizobia in Soil as Affected by Soil Temperature and Moisture

Successful inoculation of peanuts and cowpeas depends on the survival of rhizobia in soils which fluctuate between wide temperature and moisture extremes. Survival of two cowpea rhizobial strains (TAL309 and 3281) and two peanut rhizobial strains (T-1 and 201) was measured in two soils under three moisture conditions (air-dry, moist (−0.33 bar), and saturated soil) and at two temperatures (25 and 35°C) when soil was not sterilized and at 40°C when soil was sterilized. Populations of rhizobia were measured periodically for 45 days. The results in nonsterilized soil indicated that strain 201 survived relatively well under all environmental conditions. The 35°C temperature in conjunction with the air-dry or saturated soil was the most detrimental to survival. At this temperature, the numbers of strains T-1, TAL309, and 3281 decreased about 2 logs in dry soil and 2.5 logs in saturated soil during 45 days of incubation. In sterilized soil, the populations of all strains in moist soil increased during the first 2 weeks, but decreased rapidly when incubated under dry conditions. The populations did not decline under saturated soil conditions. From these results it appears that rhizobial strains to be used for inoculant production should be screened under simulated field conditions for enhanced survival before their selection for commercial inoculant production.     

Inoculation and Survival of Azotobacter chroococcum on Stored Wheat Seed

S ummary . The survival of Azotobacter chroococcum cultured on agar or in a peat-liquid medium and inoculated on to wheat seed was examined during seed storage of up to 40 weeks at room temperature. The 'half life' of peat-grown cells was 10–15 weeks, and that of an agar-grown inoculum 8–10 weeks, on seed stored in plastic bags. This was reduced to 4 weeks when agar-grown cells were stored on seed open to the air. The age of agar-grown culture between 7 and 40 days did not affect survival for up to 20 weeks. The higher numbers of Azotobacter /seed obtained with agar-grown cells outweighed the advantage gained by the slightly better survival rate of peat-grown cells.     

Seed Rain and Seed Bank Reveal that Seed Limitation Strongly Influences Plant Community Assembly in Grasslands

Dispersal is an important factor in plant community assembly, but assembly studies seldom include information on actual dispersal into communities, i.e. the local propagule pool. The aim of this study was to determine which factors influence plant community assembly by focusing on two phases of the assembly process: the dispersal phase and the establishment phase. At 12 study sites in grazed ex-arable fields in Sweden the local plant community was determined and in a 100-m radius around the centre of each site, the regional species pool was measured. The local seed bank and the seed rain was explored to estimate the local propagule pool. Trait-based models were then applied to investigate if species traits (height, seed mass, clonal abilities, specific leaf area and dispersal method) and regional abundance influenced which species from the regional species pool, dispersed to the local community (dispersal phase) and which established (establishment phase). Filtering of species during the dispersal phase indicates the effect of seed limitation while filtering during the establishment phase indicates microsite limitation. On average 36% of the regional species pool dispersed to the local sites and of those 78% did establish. Species with enhanced dispersal abilities, e.g. higher regional abundance, smaller seeds and dispersed by cattle, were more likely to disperse to the sites than other species. At half the sites, dispersal was influenced by species height. Species establishment was however mainly unlinked to the traits included in this study. This study underlines the importance of seed limitation in local plant community assembly. It also suggests that without information on species dispersal into a site, it is difficult to distinguish between the influence of dispersal and establishment abilities, and thus seed and microsite limitation, as both can be linked to the same trait.     

根瘤菌多相分类的研究进展

根瘤菌可以入侵豆科植物形成根瘤或茎瘤,并可固定空气中的氮为其宿主植物提供必需的氮素营养。根瘤菌的分类系统可分为早期根瘤菌的分类系统和现代根瘤菌的分类系统两个阶段。根瘤菌的多相分类技术包括表型的分群方法和遗传型的分群方法两大类。     

Effects of Carrier and Temperature on Survival of Rhizobium spp. in Legume Inocula: Development of an Improved Type of Inoculant

The effects of inoculant carrier, temperature, and storage period on the survival of Rhizobium strains were determined by plate count and most-probable-number analyses. Preliminary experiments showed that survival of rhizobia was affected by each of these factors and their interactions. Results of further studies indicated that six strains of rhizobia survived better at high temperatures when lyophilized and suspended in an oil carrier as compared to finely ground peat. The oil base inocula contained ca. 10⁵ viable rhizobia per g after 56 days of incubation at 60°C, whereas peat base inocula contained ≤10 rhizobia per g. These results suggest that an oil carrier will protect rhizobia from rapid death at usually lethal high temperatures.     

Carotenoids of Rhizobia

With increasing concentrations in the growth medium of the cyclization inhibitors nicotine or 2-(4-chlorophenylthio)-triethylamine hydrochloride (CPTA) the previously identified bicyclic carotenoids of Rhizobium lupini (2,3,2,3-tetrahydroxy-,-caroten-4-one and 2,3,2,3-tetrahydroxy-,-carotene) were successively replaced by hitherto unknown monocyclic carotenoids. By application of mass and nuclear magnetic resonance spectroscopy 3 carotenoids were identified as 2,3-trans-dihydroxy-,-caroten-4-one, 2,3-trans-dihydroxy-,-carotene, and 3-hydroxy-,-caroten-4-one. A further compound was tentatively established as (2- or 3-)monohydroxy-,-carotene. It was found that other inhibitors such as diphenylamine or 4-chloro-5-(dimethylamino)-2-,,(trifluoro-m-tolyl)-3-(2H)-pyridazinone (San 6706) did not affect the pigment pattern. The results are discussed in relation to carotenoid biosynthesis in Rhizobium lupini.Abbreviations CPTA 2-(4-chlorophenylthio)-triethylamine hydrochloride - San 6706 4-chloro-5-(dimethylamino)-2-,,-(trifluoro-m-tolyl)-3-(2H)-pyridazinone     

Seed Maturity and the Effects of Different Drying Conditions on Desiccation Tolerance and Seed Longevity in Foxglove (Digitalis purpurea L.)

The effects of different drying methods on desiccation toleranceand longevity in seeds of foxglove (Digitalis purpurea L.) wereassessed from just prior to mass maturity (when seeds have attainedmaximum dry weight), and at intervals during the post-abscissionphase of development. Tolerance of drying under seed conservationconditions (15% relative humidity, RH, and 15 °C), was acquiredclose to mass maturity at 36 d after flowering (DAF). Increasesin desiccation tolerance were induced when drying was delayedfor 4 d by placing seeds in a near-saturated atmosphere (approx.100% RH), or if seeds were pre-dried for 7 d at either approx.32% or approx. 73% RH. Irrespective of the drying treatment, seed longevity increasedthroughout the sampling period, i.e. beyond the point of massmaturity and throughout the post-abscission phase, up to thepoint of incipient natural dispersal. At each developmentalstage, delayed drying or pre-drying led to an increase in seedlongevity under controlled ageing conditions compared with seedsdried directly under seed conservation conditions. Increasesin longevity were apparent as increases in the estimates forthe intercept of transformed seed survival curves (K_i) and forthe standard deviation of the normal distribution of seed lifespans,and also in the mean time to death of individuals in storage,consistent with a continuation of ripening events. The results are discussed in relation to the assessment of seedlongevity and to current post-harvest drying practices for seedsintended for long-term ex-situ conservation.Copyright 1995,1999 Academic Press Digitalis purpurea L., foxglove, seed development, seed drying, seed longevity     

Weed Seed Survival during Anaerobic Digestion in Biogas Plants

Anaerobic digestion using animal manure and crop biomass is increasingly being used to produce biogas as a durable alternative to fossil fuel. The sludge, the leftover after processing, is returned to the field as a crop fertilizer. If weed seeds survive anaerobic digestion, the use of contaminated sludge poses a phytosanitary risk. The conditions that seeds are likely to encounter in biogas plants, and the effect of these, in particular temperature, on seed viability were reviewed. Knowledge on seed defence mechanisms and how these might protect seeds from inactivation in biogas reactors was summarized. Mechanisms of seed inactivation can be classified as thermal, biological and chemical. Weed species with hard seeds (physical dormant), high thermoresistance, a thick seed coat or adapted to endozoochory were identified as high-risk species. Specific seed traits could be used in future tests to circumvent extensive testing of seeds in biogas reactors.     

Growth of Fast- and Slow-Growing Rhizobia on Ethanol

Free-living soybean rhizobia and Bradyrhizobium spp. (lupine) have the ability to catabolize ethanol. Of the 30 strains of rhizobia examined, only the fast- and slow-growing soybean rhizobia and the slow-growing Bradyrhizobium sp. (lupine) were capable of using ethanol as a sole source of carbon and energy for growth. Two strains from each of the other Rhizobium species examined (R. meliloti, R. loti, and R. leguminosarum biovars phaseoli, trifolii, and viceae) failed to grow on ethanol. One Rhizobium fredii (fast-growing) strain, USDA 191, and one (slow-growing) Bradyrhizobium japonicum strain, USDA 110, grew in ethanol up to concentrations of 3.0 and 1.0%, respectively. While three of the R. fredii strains examined (USDA 192, USDA 194, and USDA 205) utilized 0.2% acetate, only USDA 192 utilized 0.1% n-propanol. None of the three strains utilized 0.1% methanol, formate, or n-butanol as the sole carbon source.     

Variability in Migration Routes Influences Early Marine Survival of Juvenile Salmon Smolts

Variability in animal migratory behavior is expected to influence fitness, but few empirical examples demonstrating this relationship exist. The initial marine phase in the migration of juvenile salmon smolts has been identified as a potentially critical life history stage to overall population productivity, yet how fine-scale migration routes may influence survival are unknown. Large-scale acoustic telemetry studies have estimated survival rates of outmigrant Pacific salmon smolts through the Strait of Georgia (SOG) along the British Columbian coastline to the Pacific Ocean, but these data have not been used to identify and characterize fine-scale movements. Data collected on over 850 sockeye salmon (Oncorhynchus nerka) and steelhead (Oncorhynchus mykiss) smolts detected at an array in the Strait of Georgia in 2004–2008 and 2010–2013 were analyzed to characterize migration routes and link movements to subsequent survival at an array 250 km further along the marine migration pathway. Both species exhibited disproportionate use of the most eastern route in the Strait of Georgia (Malaspina Strait). While many smolts moved across the northern Strait of Georgia acoustic array with no indication of long-term milling or large-scale east-to-west movements, large proportions (20–40% of sockeye and 30–50% of steelhead) exhibited a different behavior, apparently moving in a westward or counterclockwise pattern. Variability in migratory behavior for both species was linked to subsequent survival through the Strait of Georgia. Survival for both species was influenced by initial east-to-west location, and sockeye were further influenced by migration timing and duration of time spent near the northern Strait of Georgia array. Westward movements result in a net transport of smolts from Malaspina Strait to the Strait of Georgia, particularly for steelhead. Counterclockwise movements may be due to the currents in this area during the time of outmigration, and the higher proportion of steelhead smolts exhibiting this counterclockwise behavior may reflect a greater exposure to wind-altered currents for the more surface-oriented steelhead. Our results provide an empirical example of how movements can affect migration survival, for which examples remain rare in movement ecology, confirming that variability in movements themselves are an important part of the migratory process.     

Sensitivity of Rhizobia to vanillin

The sensitivity of Rhizobia isolated from pea, fenugreek, gram, cowpea, urid andRhizobium meliloti, Rhizobium trifolii andRhizobium japonicum was studied in asparagine-mannitol, salt-mannitol and salt media. It was observed that Rhizobia from gram were most resistant.Rhizobium japonicum was found to be resistant up to 50 mg vanillin/100 ml. Vanillin-resistant strains could utilize vanillin as the sole carbon source. In the case of pea, gram and fenugreek vanillin was found to be utilized in the presence of mannitol.     

影响根瘤菌竞争结瘤的生态学因素分析

根瘤菌的发现并确证其共生固氮作用已逾100年,根瘤菌剂的制备和应用也已超过半个世纪,实践效果有目共睹。如美国对豌豆根瘤菌、三叶草根瘤菌和大豆根瘤菌的应用以及澳大利亚对三叶草根瘤菌的应用都取得显著成绩。我国在豆科作物和豆科绿肥上应用根瘤菌接种措施已有30余年历史,采用筛选的优良菌     

大豆根瘤菌对福美双杀菌剂抗性的研究

通过不同大豆根瘤菌对福美双抗性的研究证明:不同大豆根瘤菌自身固有的对福美双的抗性不同,一般可耐50～100μg/ml浓度的福美双。种子拌0.3％福美双同时拌根瘤菌会使种子上的根瘤菌数降低。本文探讨了利用自发突变选育双抗菌种的可能性。试验还证明:对福美双抗性低的菌种产生自发突变抗福美双的机率要多。