Polyphasic characterization of Brazilian Rhizobium tropici strains effective in fixing N2 with common bean (Phaseolus vulgaris L.)

Common bean (Phaseolus vulgaris) is native to the Americas, and Rhizobium etli is the dominant microsymbiont in both the Mesoamerican and the Andean centers of genetic diversification. Wild common beans are not found in Brazil, although the legume has been cropped in the country throughout time and all but one of the rhizobial species that nodulate it (Rhizobium gallicum) have been broadly detected in Brazilian soils. However, the majority of the effective rhizobial strains isolated so far from field-grown plants belong to R. tropici. This study describes the analysis of symbiotic and non-symbiotic genes of 15 effective R. tropici strains, isolated from four geographically distant regions in Brazil. With RFLP-PCR of the 16S and 23S rRNA genes and sequence analysis of 16S rRNA, two clusters were observed, one related to R. tropici type A and another to type B strains. Diversity in ribosomal genes was high, indicating that type A strains might represent a new species. High intraspecies diversity was also observed in the rep-PCR analysis with BOX, ERIC and REP primers. However, in the RFLP-PCR analysis of nifH and nodC genes, all R. tropici showed unique combinations of profiles, which might reflect an evolutionary strategy to maximize N₂ fixation.     

Campylobacter jejuni, Campylobacter coli, and cytolethal distending toxin (CDT) genes in common teals (Anas crecca)

To evaluate the presence of Campylobacter spp. and related cdt genes, cloacal swabs were collected from 70 common teals (Anas crecca) and analyzed by culture methods and polymerase chain reaction. In addition, C. jejuni were examined also for the presence of wlaN gene. This is believed to be the first report of Campylobacter spp. in common teal and our results confirm the very common occurrence of C. jejuni (n=40) and C. coli (n=13) in waterfowls. Furthermore, the cdt genes were frequently present in both C. jejuni and C. coli isolated. Moreover, seven C. jejuni isolates carried also the wlaN gene which is presumably involved in the expression of ganglioside mimics in Guillain-Barré syndrome.     

Modelling diapause termination and phenology of the Japanese beetle,Popillia japonica

We deve?loped a mechanistic, stage-structured model simulating the phenology of Popillia japonica. The model simulates the influence of soil temperature on the larval diapause termination and on the development rate function of post-overwintering larvae and pupae. Model parameters are estimated based on literature evidence for pupae development and on a parameterisation process that allows estimating parameters for larval diapause termination and for the development rate function (and the related uncertainty) of post-overwintering larvae. Data used for model parameterisation and validation refer to time-series adult trap catches collected during the P. japonica monitoring programme performed by the Phytosanitary Service of Lombardy Region within the infested area in Lombardy (Italy) from 2015 to 2019. A total of 12 randomly selected locations are used to estimate biologically realistic model parameters (parameterisation dataset). We applied a Jackknife nonparametric resampling procedure on the parameterisation dataset to quantify uncertainty associated with parameters’ estimates. Parameterised model is then validated on time-series adult trap catches data referring to a different set of 12 randomly selected locations (validation dataset) surveyed in Lombardy. The model successfully predicted the beginning of adult emergence and the overall curve of adult emergence in the validation dataset. The model presented can support the definition of the best timing for the implementation of monitoring and control activities for the local and the area-wide management of P. japonica.

     

Diversity and symbiotic effectiveness of rhizobia isolated from field-grown soybean nodules in Paraguay

Soybean was introduced in Paraguay in the 1920s and commercial crops have been grown since the 1970s. Root nodulation occurs at the majority of the producing sites, although inoculation has been practiced in only 15-20% of the cropping areas. The diversity and symbiotic effectiveness of soybean rhizobia was studied using 78 isolates obtained from root nodules of field-grown plants at 16 sites located in the two main producing states. The rhizobial isolates were characterized in relation to several parameters in vitro (colony morphology, tolerance to high temperature and salinity, intrinsic resistance to antibiotics, synthesis of indole acetic acid, profiles of proteins and lipopolysaccharides) and in vivo (nodulation, plant growth and total N accumulated in shoots). Fifty-eight isolates had slow growth rates and alkaline reaction in medium containing mannitol as the carbon source, whereas 20 had fast growth rates and an acid reaction. Most isolates did not tolerate acidity (pH 4.5) or high temperature (40°C). Very few isolates shared similar protein and lipopolysaccharide profiles; therefore a high level of diversity was detected, with most of the isolates representing unique strains. Some of the isolates with an outstanding symbiotic performance were identified, and will now be tested under field conditions in a search for efficient and competitive strains for use in commercial inoculants in Paraguay.     

Effects of different management practices on arbuscular mycorrhizal fungal diversity in maize fields by a molecular approach

As obligate mutualistic symbionts, arbuscular mycorrhizal fungi (AMF) colonize the roots of many agricultural crops, and it is often claimed that agricultural practices are detrimental to AMF. As a result, agroecosystems impoverished in AMF may not get the fully expected range of benefits from these fungi. Using molecular markers on DNA extracted directly from soil and roots, we studied the effects of different management practices (tillage and N fertilization) on the AMF communities colonizing an experimental maize field in Central Italy. Our molecular analysis based on three different nuclear rRNA regions (18S, 28S and ITS) allowed us to assess AMF biodiversity. Glomeraceae members were the main colonizer, and they co-occurred with Gigasporaceae and Paraglomus regardless of the management practices applied. Diversisporaceae and Entrophosporaceae members were instead detected in the N-fertilized soils and in the untreated soil, respectively. The results obtained indicated that the general AMF assemblages structure and composition in the maize field plots appear to be primarily influenced by N fertilization and, to a lesser extent, by tillage. This study also validates the usefulness of multiple molecular markers to consolidate and refine the assessment of the environmental AMF diversity.     

Antioxidant activity and phenolic composition of wild, edible, and medicinal fennel from different Mediterranean countries

Fennel (Foeniculum vulgare Mill.) is a typical aromatic plant of the Mediterranean area, long used as a medicinal and spice herb. Fennel is also well-known for its essential oil, which has been extensively studied for many years owing to its commercial importance. In this work, the antioxidant activity and the total phenolic and flavonoid contents, as well as the quantitative determination of individual flavonoids and phenolic acids of wild, edible, and medicinal fennel from different Mediterranean countries, have been determined. The antioxidant activity was measured as the free radical (DPPH), hydroxyl radical, and superoxide anion scavenging activities. Wild fennel was found to exhibit a radical scavenging activity, as well as a total phenolic and total flavonoid content, higher than those of both medicinal and edible fennels.     

Response of field-grown bean (Phaseolus vulgaris L.) to Rhizobium inoculation and nitrogen fertilization in two Cerrados soils

 Most soils sown with field beans (Phaseolus vulgaris L.) contain indigenous rhizobia which might interfere with the establishment of inoculated strains. As a consequence, the benefits of bean inoculation are usually questioned, and the use of N fertilizer is gradually becoming a common practice. The present study had the objective of evaluating the effectiveness of inoculation and N fertilization in field soil with (site 1) and without (site 2) a previous bean-cropping history. At site 1, which had a rhizobial population of 7×10² cells g^–1 soil, inoculation had no effect on nodulation or yield, whereas at site 2 (<10 cells g^–1 soil) inoculation increased nodulation, nodule occupancy by the inoculated strain and grain yield. N fertilizer decreased nodulation at both sites, but increased grain yield at site 1 but not at site 2, indicating that the response to inoculation and N fertilization depends on the cropping history. When bean was cultivated for the first time, indigenous populations of rhizobia were low and high yields were accomplished solely with seed inoculation, with no further response to N fertilizer. In contrast, previous cultivation of bean increases soil rhizobia, preventing nodule formation by inoculated strains, and N fertilizer may be necessary for maximum yields. A significant interaction effect between N fertilizer and inoculation was detected for serogroup distribution only at site 2, with N fertilizer decreasing nodule occupancy by the inoculated strain and increasing the occurrence of indigenous strains. Consequently, although no benefits were obtained by the combination of inoculation and N fertilizer, this practice may be feasible with the selection of appropriate N-tolerant strains from the indigenous rhizobial population. Received: 26 May 1999     

Are the rates of photosynthesis stimulated by the carbon sink strength of rhizobial and arbuscular mycorrhizal symbioses?

Rhizobial and arbuscular mycorrhizal (AM) symbioses each may consume 4-16% of recently photosynthetically-fixed carbon to maintain their growth, activity and reserves. Rhizobia and AM fungi improve plant photosynthesis through N and P acquisition, but increased nutrient uptake by these symbionts does not fully explain observed increases in the rate of photosynthesis of symbiotic plants. In this paper, we test the hypothesis that carbon sink strength of rhizobial and AM symbioses stimulates the rates of photosynthesis. Nutrient-independent effects of rhizobial and AM symbioses result in direct compensation of C costs at the source. We calculated the response ratios of photosynthesis and nutrient mass fraction in the leaves of legumes inoculated with rhizobial and/or AM fungi relative to non-inoculated plants in a number of published studies. On average, photosynthetic rates were significantly increased by 28 and 14% due to rhizobial and AM symbioses, respectively, and 51% due to dual symbiosis. The leaf P mass fraction was increased significantly by 13% due to rhizobial symbioses. Although the increases were not significant, AM symbioses increased leaf P mass fraction by 6% and dual symbioses by 41%. The leaf N mass fraction was not significantly affected by any of the rhizobial, AM and dual symbioses. The rate of photosynthesis increased substantially more than the C costs of the rhizobial and AM symbioses. The inoculation of legumes with rhizobia and/or AM fungi, which resulted in sink stimulation of photosynthesis, improved the photosynthetic nutrient use efficiency and the proportion of seed yield in relation to the total plant biomass (harvest index). Sink stimulation represent an adaptation mechanism that allows legumes to take advantage of nutrient supply from their microsymbionts without compromising the total amount of photosynthates available for plant growth.     

Characterization of Neofusicoccum parvum causing canker and dieback on Brachychiton species

Brachychiton species are planted in Italy as ornamental trees. A survey in a nursery in Sicily (Italy) revealed the presence of young B. acerifolius and B. populneus showing severe trunk cankers, massive gummosis from the bark cracks, wood discolouration, and twig dieback. Morphological characterization was based on conidia measurement, and molecular characterization was carried out performing phylogenetic analysis (Maximum Parsimony) based on multi-locus approach of partial ITS, EF-1α, and tub2. Identification of the causal agent resulted in Neofusicoccum parvum (Botryosphaeriaceae). Pathogenicity tests were conducted in order to fulfill the Koch’s postulates. Five and three-years-old potted Brachychiton plants were inoculated (indoor and outdoor) using mycelial plugs. All the inoculated plants showed severe symptoms similar of those observed in the nursery during the survey. Some of the inoculated plants were completely dead. Re-isolations fully confirmed the causal agent. For our knowledge, this is the first report worldwide of N. parvum attacking Brachychiton spp.