Genetic Effects on Host × Strain Interaction in the Symbiosis of Vicia faba and Rhizobium leguminosarum

50 Vicia faba lines were inoculated with 4 strains of Rhizobium leguminosarum in all possible combinations under greenhouse conditions in 5 replications. A root-sterile method was used for the cultures. Characters such as plant height, leaf colour, number and colour of nodules, the dry weights of shoot, root and nodules, and the nitrogen content were recorded, after 10 weeks. It was demonstrated by analysis of variance that, for example, for the dry matter of the shoot only 1.9 % of the total phenotypic variance was caused by interaction between Vicia and Rhizobium genotypes, but 24.7 % Was conditioned by the faba bean genotypes and 73.4 % by the Rhizobium strains. Differences of Vicia faba genotypes in their reaction to different Rhizobium strains were detected by joint regression analysis.     

Symbiotic variability in Vicia faba. 1. Genetic variation in the Rhizobium leguminosarum population

Summary Twenty isolates taken at random from indigenous populations of Rhizobium leguminosarum sampled near Aberystwyth were each inoculated into plants of a standard Vicia faba variety grown aseptically under conditions free of combined nitrogen. Plants in association with the individual rhizobium isolates exhibited large differences in dry matter yield, nitrogen content, efficiency of nitrogen utilization and date of first flower. The implications of these differences to the productivity and reliability of varieties of field beans are discussed.     

Influence of acid soil on nodule numbers in relation to polyamine and tannin concentrations in roots of Phaseolus vulgaris

The relationship between soil acidity and polyamine (putrescine, spermidine, spermine) concentrations in roots was evaluated and compared to (1) nodule numbers in common beans and (2) tannin concentrations in roots. Six Phaseolus vulgaris cultivars were grown in pots in a greenhouse in soil at pH 4.5 or 5.5 and inoculated with Rhizobium leguminosarum bv. phaseoli strains. At pH 4.5 nodule numbers were strongly reduced but polyamine concentrations were 50% higher than at pH 5.5. At both pH levels putrescine and spermidine were the dominant polyamines, while the concentration of spermine was less than half of the other two. There was also a cultivar effect on the polyamine concentration, but this was much less pronounced than the pH effect. By 22 days after planting the uninoculated control plants had about 13% lower levels of polyamines than the inoculated plants. The concentrations of putrescine, spermidine, and spermine were negatively correlated with nodule numbers, but positively correlated with the root tannin content. There were no significant correlations with either root or plant dry weight.     

Structure and diversity among rhizobial strains, populations and communities-a review

Published studies of rhizobial populations, communities and other strain collections were analysed to identify trends in strain richness, strain dominance and genetic diversity within and between locations. For individual populations and communities, strain richness indices were calculated by dividing the number of strain types identified by the number of isolates recovered. Where possible, strain dominance (the proportion of rhizobial isolates represented by each strain type) was also calculated. Analysis of the genetic diversity of populations, communities and rhizobial strain collections originating from diverse legume hosts and locations, was confined to studies using multilocus enzyme electrophoresis (MLEE) so that diversity could be compared on the basis of published H values. Strain richness indices were highly variable (0.02-0.94) and were influenced by both the discriminatory power of the strain typing method and the type and number of legume species used to recover rhizobia from soil. The strain richness of populations recovered either directly from soil, or from the nodules of trap hosts inoculated with the same soil, was similar. Because the arithmetic relationship between the number of strain types and number of isolates varies between different populations and communities, strain richness curves are proposed as the most appropriate method for reporting rhizobial structural diversity. Comparison of over 50 rhizobial populations and communities from published studies showed that strain dominance patterns in nodules were often similar. Typically, a single strain type occupies more than 30% of nodules with the majority of strain types being recovered at low frequency (∼75% of published populations and communities). Rhizobial populations and communities characterised by MLEE varied in their genetic diversity, with H values ranging from 0.06 to 0.78. In several studies, most of the genetic diversity within a site could be recovered from the nodules of a single plant. When hierarchical analyses were performed on populations within and between sites, the genetic diversity within sites was similar to the genetic diversity between sites. Similarly, the genetic diversity of strain collections originating from multiple hosts and locations was no more diverse than some individual populations and communities. Strain richness and genetic diversity measures were not always correlated for rhizobial populations. Populations with low strain richness were sometimes genetically diverse, and the relationship between the diversity measures varied for different legume species at the same location. We suggest that both strain richness and genetic diversity measures are required to fully describe rhizobial population and community diversity.     

Pesticide toxicity using protozoans as test organisms

Summary A toxicity test using protozoa as test organisms was devised for pesticides. Two different methods were used, the most probable number method and the absorbance method (based on measurement of protozoa growth through absorbance). The ciliates Colpoda cucullus and Blepharisma undulans and the flagellate Oikomonas termo were isolated from different biological starter cultures and tested with the herbicides Chlorex, MCPA, dichlorprop and Matrigon, the fungicide Benlate, and the insecticide Sumicidin. The protozoans showed quite different sensitivities to the pesticides, using 9-h lethal concentrations (LC₅₀ and LC₁₀) as criteria. The 9-h LC₅₀ (concentration at which 50% of the protozeon population has died after 9 h of incubation) ranged from 0.7 ppm for Benlate to 40000 ppm for Chlorex. The usual soil application rates of Chlorex, MCPA, and Benlate were toxic to some of the organisms.     

Initial results from long-term field studies at three sites on the effects of heavy metal-amended liquid sludges on soil microbial activity

In a long‐term study of the effects on soil fertility and microbial activity of heavy metals contained in sewage sludges, metal‐amended liquid sludges each with elevated Zn, Cu or Cd concentrations were applied over a 3‐year period (1995–1997) to three sites in England. The experiments were sited adjacent to experimental plots receiving metal‐rich sludge cakes enabling comparisons to be made between the effects of heavy metal additions in metal‐amended liquid sludges and sludge cakes. The liquid sludge additions were regarded as ‘worst case’ treatments in terms of likely metal availability, akin to a long‐term situation following sewage sludge additions where organic matter levels had declined and stabilised. The aim was to establish individual Zn (50–425 mg kg^?1), Cu (15–195 mg kg^?1) and Cd (0.3–4.0 mg kg^?1) metal dose–response treatments at each site, but with significantly smaller levels of organic matter addition than the corresponding sludge cake experiments. There were no differences (P > 0.05) in soil respiration rates, biomass carbon concentrations or most probable numbers of clover Rhizobium between the treatments at any of the sites at the end of the liquid sludge application programme. Soil heavy metal extractability differed between the metal‐amended liquid sludge and metal‐rich sludge cake treatments; Zn and Cd extractabilities were higher from the liquid sludge additions, whereas Cu extractability was higher from the sludge cake application. These differences in metal extractability in the treated soil samples reflected the contrasting NH₄NO₃ extractable metal contents of the metal‐amended liquid sludges and sludge cakes that were originally applied.     

Infection and colonization of aseptically micropropagated sugarcane seedlings by nitrogen-fixing endophytic bacterium, Herbaspirillum sp. B501gfp1

In this study, we used Herbaspirillum sp. B501gfp1 (B501gfp1), an isolate from wild rice, to investigate the interaction between a non-host nitrogen-fixing endophytic bacterium and micropropagated sugarcane plants under aseptic condition. Two Japanese sugarcane plants (Saccharum sp.) cultivars (cvs) NiF8 and Ni15 were inoculated using B501gfp1 in two inoculum doses of 10⁸ and 10² bacterial-cells-per-milliliter suspension. The results showed that bacterial cells colonized both the root and stem tissues, and colonization was apparent in the intercellular spaces. Higher bacterial numbers were detected in plant tissues inoculated with the higher inoculum concentration treatment. Bacterial numbers also varied between the two cultivars, with the higher values determined in cv Ni15. This study provides evidence that Herbaspirillum sp. B501gfp1, a rice isolate, could colonize sugarcane tissues, suggesting non-specificity of host plant among endophytes.     

Evaluation of the intrinsic competitiveness and saprophytic competence of Rhizobium tropici IIB strains

Inoculation of beans (Phaseolus vulgaris L.) with strains of R. tropici IIB and R. etli resulted in the disappearance of the R. tropici IIB stains from the nodule population and their replacement by other (non R. tropici IIB) bean symbionts (Vlassak et al. 1996). Coinoculation studies in monoxenic conditions and in soil core microcosms with plants harvested at two different growth stages indicated that the inoculated R. tropici IIB strains CIAT899 and F98.5 possess a good intrinsic competitiveness which declines, however, at a later plant growth stage and in soil conditions. The poor saprophytic competence of R. tropici IIB strain CIAT899 was further demonstrated by its poor survival in soil core microcosms after bean harvest. Strains were isolated from the field plots with a 3-year bean-planting history, characterized and evaluated for their competitiveness against R. tropici IIB strain CIAT899. Isolates from field plots, which had been repeatedly inoculated with R. tropici IIB strain CIAT899, showed a higher nodule occupancy compared to R. tropici IIB strain CIAT899, and this higher competitiveness exhibited by the field isolates might be an additional reason for the poor performance of R. tropici IIB strain CIAT899 in the field study. Plots with and without a history of bean production revealed after 3-year bean cultivation an almost totally different population that also significantly differed in competitiveness. Received: 12 February 1996     

国槐、剌槐幼苗根系固氮酶活性的研究

研究了刺槐(Robinia pseudoacacia)、国槐(Sophora japonica)一年生幼苗根系固氮酶活性的季节性变化,并且通过植物离体根尖接种根瘤菌试验和分析植物根系分泌物反映了国槐、刺槐根系固氮的某些特性。     

硒对小麦生理功能的影响之研究进展

为获得能够使紫花苜蓿在酸性土壤中高效结瘤的耐酸根瘤菌,该研究从酸性土(pH5.2)上生长的紫花苜蓿根瘤中分离到若干菌株,利用根瘤菌对酸的适应性耐受反应筛选到六株能够在pH4.8的固体培养基上稳定生长的菌株,并检测了这六株菌的结瘤特征、不同酸度下的生长曲线和平均代时。结果表明,六株根瘤菌均能使紫花苜蓿植株结瘤,使其具有较多的结瘤数(≥10个/株)和较高的结瘤率(66.67%～100%);不同酸度下,六株根瘤菌呈现不同的生长特性,酸显著降低根瘤菌的生长速率,使其生长延滞期增长,对数生长期出现滞后,稳定期到达延迟,酸性条件下各根瘤菌的平均代时均较中性条件下的长,环境pH越低,菌株平均代时越长。然而,在低pH条件下,耐酸根瘤菌在其耐受酸度范围内经过延长的延滞期后仍然能够较快地生长。