Maria Rita Scotti

Nodulation and nitrogen fixation by Mimosa spp. in the Cerrado and Caatinga biomes of Brazil

*An extensive survey of nodulation in the legume genus Mimosa was undertaken in two major biomes in Brazil, the Cerrado and the Caatinga, in both of which there are high degrees of endemicity of the genus. *Nodules were collected from 67 of the 70 Mimosa spp. found. Thirteen of the species were newly reported as nodulating. Nodules were examined by light and electron microscopy, and all except for M. gatesiae had a structure typical of effective Mimosa nodules. The endosymbiotic bacteria in nodules from all of the Mimosa spp. were identified as Burkholderia via immunolabelling with an antibody against Burkholderia phymatum STM815. *Twenty of the 23 Mimosa nodules tested were shown to contain nitrogenase by immunolabelling with an antibody to the nitrogenase Fe- (nifH) protein, and using the delta(15)N ((15)N natural abundance) technique, contributions by biological N(2) fixation of up to 60% of total plant N were calculated for Caatinga Mimosa spp. *It is concluded that nodulation in Mimosa is a generic character, and that the preferred symbionts of Brazilian species are Burkholderia. This is the first study to demonstrate N(2) fixation by beta-rhizobial symbioses in the field.

Dual inoculation of a woody legume (Centrolobium tomentosum) with rhizobia and mycorrhizal fungi in south-eastern Brazil

The integration of N2 fixing trees into stable agroforestry systems in the tropics is being tested due to their ability to produce high biomass N and P yields, when symbiotically associated with rhizobia and mycorrhizal fungi. The growth of Centrolobium tomentosum Guill. ex Benth, a native leguminous tree from the Brazilian Atlantic Forest, was assessed with dual inoculation of Rhizobium spp and mycorrhizal fungi under field conditions. Complete fertilization was compared to treatments of inoculation with selected rhizobia strains BHICB-Ab1 or BHICB-Ab3, associated or not to arbuscular mycorrhizal (AM) fungi. The dual inoculation increased the height and growth in relation to the plants treated with rhizobia alone. Plants inoculated with strain BHICB-Ab1 and arbuscular mycorrhizal fungi (AMF) exhibited an increase of 56% dry matter over uninoculated control and nitrogen accumulation was greater than with BHICB-Ab3 inoculated plants. Strain BHICB-Ab1 presented a synergetic relationship with mycorrhizal fungi since the combined inoculation with BHICB-Ab1 enhanced plant height and dry weight more than single inoculation while the growth of BHICB-Ab3 plants was not modified by AMF inoculation. Arbuscular mycorrhizal fungi enhanced plants survival and seemed to favour the nodule occupation by rhizobia strains as compared to the non-mycorrhizal plants. Inoculation with selected rhizobia and AMF improved the growth of C. tomentosum under field conditions.

Nodulation in Dimorphandra wilsonii Rizz. (Caesalpinioideae), a Threatened Species Native to the Brazilian Cerrado

The threatened caesalpinioid legume Dimorphandra wilsonii, which is native to the Cerrado biome in Brazil, was examined for its nodulation and N2-fixing ability, and was compared with another, less-threatened species, D. jorgei. Nodulation and potential N2 fixation was shown on seedlings that had been inoculated singly with five bradyrhizobial isolates from mature D. wilsonii nodules. The infection of D. wilsonii by two of these strains (Dw10.1, Dw12.5) was followed in detail using light and transmission electron microscopy, and was compared with that of D. jorgei by Bradyrhizobium strain SEMIA6099. The roots of D. wilsonii were infected via small transient root hairs at 42 d after inoculation (dai), and nodules were sufficiently mature at 63 dai to express nitrogenase protein. Similar infection and nodule developmental processes were observed in D. jorgei. The bacteroids in mature Dimorphandra nodules were enclosed in plant cell wall material containing a homogalacturonan (pectic) epitope that was recognized by the monoclonal antibody JIM5. Analysis of sequences of their rrs (16S rRNA) genes and their ITS regions showed that the five D. wilsonii strains, although related to SEMIA6099, may constitute five undescribed species of genus Bradyrhizobium, whilst their nodD and nifH gene sequences showed that they formed clearly separated branches from other rhizobial strains. This is the first study to describe in full the N2-fixing symbiotic interaction between defined rhizobial strains and legumes in the sub-family Caesalpinioideae. This information will hopefully assist in the conservation of the threatened species D. wilsonii.

Effect of post-planting inoculation with Bradyrhizobium sp and mycorrhizal fungi on the growth of Brazilian rosewood, Dalbergia nigra Allem. ex Benth., in two tropical soils

Six months after planting, seedlings of the leguminous treeDalbergia nigra, known as Brazilian rosewood, wereinoculated with a selected Bradyrhizobium strainBHICB-DN 15,either alone or in combination with mycorrhizal fungi to determine their effecton the growth of D. nigra in Atlantic forest andeucalyptussoils. Height growth of D. nigra was similar in both soilsand six months after inoculation Bradyrhizobium did notaffect the D. nigra height growth, but it did improve drymass and especially in the nitrogen content of plants grown in eucalyptus soil.The success of the delayed inoculation with BHICB-DN 15 confirms its competitiveability “vis-à-vis” indigenous soil rhizobia. Co-inoculationof BHICB-DN 15 and mycorrhizal fungi did not increase plant dry weight,nitrogenand phosphorus content or mycorrhizal colonization. These results suggest thatthe BHICB-DN 15 strain had no synergistic relationship with the mycorrhizalfungi or that there was incompatibility between symbionts, in both soils.

Effect of the inoculation and distribution of mycorrhizae in Plathymenia reticulata Benth under monoculture and mixed plantation in Brazil

This paper investigates the distribution of arbuscular mycorrhizal fungi (AMF) spores and AMF colonization in a field study in southeastern Brazil. Response to AMF and rhizobial inoculation was studied in monocultures of Plathymenia reticulata and mixed plantations with both Tabebuia heptaphylla and Eucalyptus camaldulensis in a sandy soil during two consecutive years. P. reticulata height and diameter and mycorrhizal colonization and AMF diversity were measured in dry and rainy periods. The inoculated treatment of E. camaldulensis, T. heptaphylla and P. reticulata mixed plants showed higher height and diameter growth of P. reticulata used as well as increased root colonization and AMF spore numbers. Spore populations were found to belong to five genera: Acaulospora, Entrophospora, Glomus, Gigaspora and Scutellospora, with Glomus dominating. Agroforestry practices including use of leguminous tree P. reticulata effectively maintained AMF spore numbers in soils and high AMF colonization levels compared with monocultures, proving an efficient system for productivity and sustainability.

Laboratory decomposition of Dalbergia nigra all. ex. benth and Eucalyptus grandis w.hill ex. maiden leaves in forest and eucalypt plantation soils

The objective was to evaluate the decomposition of D. nigra and E. grandis leaves under laboratory conditions when incubated in soils from Atlantic Forest and from a 40-year-old eucalypt plantation for a year. The obtained values of pH in forest and eucalypt plantation soils were 5.61 and 4.47 and for aluminum 0.32 and 1.89mEqx100, respectively. A great number of microorganisms (total fungi, phosphate solubilising and cellulolytic organisms) were found in forest soil. Litterbag study revealed a higher mass loss of the leaves in forest soil in the first four months (c. 30%). In the eucalypt plantation soils loss was observed only after eight months (c. 40%). The initial contents of N and P were greater in D. nigra than in E. grandis leaves, although lignin and cellulose contents were similar. The C/N ratio was higher in E. grandis indicating that its degradation could be slower. Nonetheless, there were no significant differences in the decay rates (k) among the treatments. Phosphorus, lignina and cellulose were lost at the end of the experiment in all treatments. The results show the differences between decomposition of both species and the influence of forest and eucalypt plantation soils in the decomposition process.

DECOMPOSITION, DYNAMIC OF INORGANIC ELEMENTS AND MICROBIAL COLONIZATION OF ARARIBÁ (Centrolobium tomentosum Guill. ex Benth) LEAVES ON LATOSOLS OF BRAZILIAN ATLANTIC FOREST

Se estan cultivando arboles de leguminosas para establecer unidades de restauracion en el ecosistema del bosque tropical atlantico de Brasil. El exito del establecimiento depende del crecimiento, productividad de las plantas, y de la tasa de descomposicion de las hojas. El objetivo de este estudio fue investigar la descomposicion de las hojas de arariba (Centrolobium tomentosum Guill. ex. Benth), en dos sitios. El Sitio 1 era una plantacion experimental de la especie nativa C. tomentosum, inoculado con rhizobios y hongos de la micorriza, y el Sitio 2 era una area con una plantacion de eucalipto (Eucalyptus grandis W. Hill ex Maiden). Las hojas caidas de C. tomentosum fueron incubadas durante 120, 240 y 360 dias en ambos sitios. La velocidad de descomposicion y la dinamica de mineralization de la materia organica y de los nutrientes K, Mg y Ca fue modificada por el efecto del sitio. La masa restante y la concentracion de nutrientes fueron mas bajas en el Sitio 2 que en el Sitio 1. La inmovilizacion del N y P ocurrio ocho meses despues de la incubacion en el Sitio 2 y cuatro meses en el Sitio 1. La colonizacion microbiana fue hecha por hongos en el Sitio 2, mientras que en el Sitio 1 fue por baterias. La alta concentracion de Cu, Mn y Zn en el Sitio 2 no tuvo en correlacion con los niveles de metales en los residuos. La concentracion de Cu y Mn en los residuos aumentaron en los dos sitios. El cambio de plantas de eucalipto a arboles de C. tomentosum modifico la biota del suelo y favorecio a las bacterias en comparacion a la poblacion de hongos.

Invasion of the Brazilian campo rupestre by the exotic grass Melinis minutiflora is driven by the high soil N availability and changes in the N cycle.

The Serra do Rola Moça State Park (PESRM) in Minas Gerais State, Brazil is a preserved site representative of the campo rupestre biome over an ironstone outcrop that has a high level of plant diversity. Almost 60% of this grassy field has been invaded by the exotic molasses grass (Melinis minutiflora), which constitutes a severe threat to the biodiversity and survival of this biome, particularly due to the impacts of annual fires and inappropriate restoration interventions. Many invasive species exhibit a high demand for nitrogen (N). Hence, this work aimed to study the N cycle alterations promoted by M. minutiflora in a site of the campo rupestre, where the leguminous species Mimosa pogocephala was prevalent. The biomes soils exhibited a high natural N fertility and low C:N ratio. The main N source in this biome resulted from the biological N fixation performed by M. pogocephala associated with Burkholderia nodosa, as evidenced by the total leaf N content, leaf δ15N signature, nodule occupation and bacterial molecular identification analyses. The displacement of native species by molasses grass was associated with changes in the soil N forms, namely the nitrate increased as the ammonium decreased. The latter was the dominant N form in the native species plots, as observed in the soil analysis of total N, ammonium and nitrate contents. The dominant ammonium form was changed to the nitric form by the stimulation of ammonia-oxidising bacteria populations due to the invasive species. Therefore, the key mechanism behind the invasiveness of the exotic grass and the concomitant displacement of the native species may be associated with changes in the soil N chemical species. Based on this finding and on the high N-based soil fertility found in the campo rupestre N fertilisation procedures for restoration of invaded areas should be strictly avoided in this biome.

Aboveground nutrient components of Eucalyptus camaldulensis and E. grandis in semiarid Brazil under the nature and the mycorrhizal inoculation conditions

A study was conducted to evaluate the aboveground biomass, nutrient content and the percentages of mycorrhizal colonization in Eucalyptus camaldulensis and Eucalyptus grandis plantations in the semiarid region (15° 09′ S 43° 49′ W) in the north of the State of Minas Gerais in Brazil. Results show that the total above-ground biomass (dry matter) was 33.6 Mg·ha−1 for E. camaldulensis and 53.1 Mg·ha−1 for E. grandis. The biomass of the stem wood, leaves, branches, and stem bark for E. camaldulensis accounted for 64.4%, 19.6%, 15.4%, and 0.6% of the total biomass, respectively (Table 2); meanwhile a similar partition of the total above-ground biomass was also found for E. grandis. The dry matter of leaves and branches of E. camaldulensis accounted for 35% of total biomass, and the contents of N, P, K, Ca, Mg, and S in leaves and branches accounted for 15.5%, 0.7%, 12.3%, 22.6%, 1.9%, and 1.4% of those in total above-ground biomass, respectively. In the trunk (bark and wood), nutrient accumulation in general was lower. Nutrient content of E. grandis presented little variation compared with that of E. camaldulensis. Wood localized in superior parts of trunk presented a higher concentration of P and bark contained significant amounts of nutrients, especially in E. grandis. This indicated that leaving vegetal waste is of importance on the site in reducing the loss of tree productivity in this semi-arid region. The two species showed mycotrophy.

Arbuscular Mycorrhizal Fungi and Arsenate Uptake by Brachiaria Grass (Brachiaria decumbens)

ABSTRACT Using a compartmentalized treatment technique, the role of arbuscular mycorrhizal fungi (AMF; Acaulospora scrobiculata) on arsenic (As) uptake and translocation in Brachiaria decumbens. Treatments consisted of a factorial arrangement of three As doses (0, 50, and 100 mg kg−1) and the presence/absence of AMF inoculates. In the absence of AMF, B. decumbens did not show As accumulation, indicating the probable presence of tolerance mechanism via As exclusion by the roots. B. decumbens plants showed high AMF colonization levels, especially in the arsenic treatments, with AMF improving shoot and root growth independent of As concentrations. Arsenic accumulation occurred only with AMF inoculation. Phosphorous uptake was reduced in B. decumbens roots in the presence of arsenic with and without inoculation of AMF. Results suggest that B. decumbens can be used in phytoremediation procedures when inoculated with A. scrobiculata, although pasture formation should be strictly avoided in contaminated sites.