Arsenic toxicity in Acacia mangium willd. and mimosa Caesalpiniaefolia benth. seedlings

Acacia mangium and Mimosa caesalpiniaefolia are fast-growing woody fabaceous species that might be suitable for phytoremediation of arsenic (As)-contaminated sites. To date, few studies on their tolerance to As toxicity have been published. Therefore, this study assessed As toxicity symptoms in A. mangium and M. caesalpiniaefolia seedlings under As stress in a greenhouse. Seedlings of Acacia mangium and M. caesalpiniaefolia were grown for 120 d in an Oxisol-sand mixture with 0, 50, 100, 200, and 400 mg kg-1 As, in four replications in four randomized blocks. The plants were assessed for visible toxicity symptoms, dry matter production, shoot/root ratio, root anatomy and As uptake. Analyses of variance and regression showed that the growth of A. mangium and M. caesalpiniaefolia was severely hindered by As, with a reduction in dry matter production of more than 80 % at the highest As rate. The root/shoot ratio increased with increasing As rates. At a rate of 400 mg kg-1 As, whitish chlorosis appeared on Mimosa caesalpiniaefolia seedlings. The root anatomy of both species was altered, resulting in cell collapse, death of root buds and accumulation of phenolic compounds. Arsenic concentration was several times greater in roots than in shoots, with more than 150 and 350 mg kg-1 in M. caesalpiniaefolia and A. mangium roots, respectively. These species could be suitable for phytostabilization of As-contaminated sites, but growth-stimulating measures should be used.

Physical-hydraulic properties of a sandy loam typic paleudalf soil under organic cultivation of 'montenegrina' mandarin (Citrus deliciosa Tenore)¹

Citrus plants are the most important fruit species in the world, with emphasis to oranges, mandarins and lemons. In Rio Grande do Sul, Brazil, most fruit production is found on small properties under organic cultivation. Soil compaction is one of the factors limiting production and due to the fixed row placement of this crop, compaction can arise in various manners in the interrows of the orchard. The aim of this study was to evaluate soil physical properties and water infiltration capacity in response to interrow management in an orchard of mandarin (Citrus deliciosa Tenore 'Montenegrina') under organic cultivation. Interrow management was performed through harrowing, logs in em "V", mowing, and cutting/knocking down plants with a knife roller. Soil physical properties were evaluated in the wheel tracks of the tractor (WT), between the wheel tracks (BWT), and in the area under the line projection of the canopy (CLP), with undisturbed soil samples collected in the 0.00-0.15, 0.15-0.30, 0.30-0.45, and 0.45-0.60 m layers, with four replicates. The soil water infiltration test was performed using the concentric cylinder method, with a maximum time of 90 min for each test. In general, soil analysis showed a variation in the physical-hydraulic properties of the Argissolo Vermelho-Amarelo distrófico arênico (sandy loam Typic Paleudalf) in the three sampling sites in all layers, regardless of the management procedure in the interrows. Machinery traffic leads to heterogeneity in the soil physical-hydraulic properties in the interrows of the orchard. Soil porosity and bulk density are affected especially in the wheel tracks of the tractor (WT), which causes a reduction in the constant rate of infiltration and in the accumulated infiltration of water in this sampling site. The use of the disk harrow and mower leads to greater harmful effects on the soil, which can interfere with mandarin production.

Behavior of Eucalyptus urophylla and Eucalyptus citriodora Seedlings Grown in Soil Contaminated by Arsenate

ABSTRACT Persistent areas of tailings and deposits from coal and gold mining may present high levels of arsenic (As), mainly in the arsenate form, endangering the environment and human health. The establishment of vegetation cover is a key step to reclaiming these environments. Thus, this study aimed to evaluate the potential of Eucalyptus urophylla and E. citriodora seedlings for use in phytoremediation programs of arsenate-contaminated areas. Soil samples were incubated at increasing rates (0, 50, 100, 200 and 400 mg dm-3) of arsenic (arsenate form, using Na2HAsO4) for 15 days. The seedlings were produced in a substrate (vermiculite + sawdust) and were transplanted to the pots with soil three months after seed germination. The values of plant height and diameter were taken during transplanting and 30, 60 and 90 days after transplanting. In the last evaluation, the total leaf area and biomass of shoots and roots were also determined. The values of available As in soil which caused a 50 % dry matter reduction (TS50%), the As translocation index (TI) from the roots to the shoot of the plants, and its bioconcentration factor (BF) were also calculated. Higher levels of arsenate in the soil significantly reduced the dry matter production of roots and shoots and the height of both species, most notably in E. urophylla plants. The highest levels of As were found in the root, with higher values for E. citriodora (ranging from 253.86 to 400 mg dm-3). The TI and BF were also reduced with As doses, but the values found in E. citriodora were significantly higher than in E. urophylla. E. citriodora plants presented a higher capacity to tolerate As and translocate it to the shoot than E. urophylla. Although these species cannot be considered as hyperaccumulators of As, E. citriodora presented the potential to be used in phytoremediation programs in arsenate-contaminated areas due to the long-term growth period of this species.

The effects of arbuscular mycorrhizal fungi inoculation on Euterpe oleracea mart. (açaí) seedlings

With the objective of verifying the response of Euterpe oleracea seedlings to seven arbuscular mycorrhizal fungi species, an experimental trial was carried out under greenhouse conditions. Seeds of E. oleracea were sown in carbonized rice husk. Germinating seeds were initially transferred to plastic cups, containing fumigated Reddish Yellow Quartz Sand and inoculated with arbuscular mycorrhizal fungi. Two months later, seedlings were transferred to 2 kg black plastic bags, containing the same soil without fumigation. Plant growth and mineral nutrients were evaluated nine months after mycorrhizal inoculation. Differential effects were observed among the species tested, with Scutellispora gilmorei being the most effective ones in promoting growth and nutrient content of E. oleracea seedlings. The increment resulted from inoculation with S. gilmorei were 92% in total plant height, 116% in stem diameter, 361% in dry matter production, 191% in N, 664% in P, 46% in K, 562% in Ca, 363% in Mg and 350% in Zn contents, comparing to uninoculated controls. Infected root length was positively correlated to nutrient content and plant growth. It was concluded that growth and nutrient uptake of E. oleracea seedlings could be significantly improved by inoculation of effective arbuscular mycorrhizal fungi.

Porta-enxertos para laranjeiras-doces (Citrus sinensis (L.) Osb.), em Rio Branco, Acre

O objetivo deste trabalho foi de avaliar o comportamento de sete cultivares de laranjeiras- doces: 'Baia 101', 'Baianinha IAC 79', 'Monte Parnaso', 'Pêra D6', 'Natal 112', 'Valência 27' e 'Aquiri', sobre diferentes porta-enxertos: limão 'Cravo', tangerinas 'Cleópatra' e 'Sunki' e citrange 'Carrizo', nas condições edafoclimáticas de Rio Branco, Acre. O delineamento experimental foi em parcelas subdivididas, com as cultivares nas parcelas, os porta-enxertos nas subparcelas e os quatro anos de avaliação como repetições. As laranjeiras 'Pêra D6', 'Natal 112' e 'Valência 27' apresentaram tendências de maior produção quando enxertadas sobre o limão 'Cravo', e a laranja 'Aquiri' quando enxertada sobre citrange 'Carrizo'. Em relação aos demais porta-enxertos, o limão 'Cravo' mostrou tendências de induzir maior produção/volume de copa e peso médio do fruto, e menor teor de sólidos solúveis totais e acidez total. As laranjas do grupo Baia ('Baia 101', 'Baianinha IAC 79' e 'Monte Parnaso') produziram frutos com baixa percentagem de suco; não são recomendadas para plantio em Rio Branco, AC. Com base nos resultados obtidos, recomendam-se os porta-enxertos citrange 'Carrizo', tangerina 'Cleópatra' e limão 'Cravo' para a laranja 'Aquiri', e o porta-enxerto limão 'Cravo' para as laranjas 'Pêra D6', 'Natal 112' e 'Valência 27'.

Efeitos de ácido giberélico e carvão ativado no cultivo in vitro de Citrus limonia Osbeck chiPoncirus trifoliata (L.) Raf

Pesquisou-se o desenvolvimento in vitro de embriões de um híbrido do cruzamento Citrus limonia Osb. chi Poncirus trifoliata (L.) Raf. em meio MS, acrescido de GA3 (0,0, 0,01, 0,1 e 1,0 mg/L) e carvão ativado (0,0, 0,5, 1,0 e 2,0 g/L), em todas as combinações possíveis. Após a excisão das sementes os embriões obtidos, independentemente dos estádios, foram inoculados nos respectivos meios de cultura. Cada tratamento permaneceu por 48 horas no escuro e 40 dias em sala de crescimento à temperatura de 27 ± 1ºC, com 16 horas de luminosidade. Após esse período, as plântulas foram avaliadas, considerando-se o porcentual de sobrevivência e comprimento da haste caulinar e do sistema radicular. Verificou-se que a concentação de 0,1 mg/L de GA3 interagiu antagonicamente em meio de cultivo contendo até 2,0 g/L de carvão ativado. O GA3 a 0,01 mg/L associado a concentrações de carvão ativado a partir de 0,5 g/L até 2,0 g/L, maximizou o porcentual de sobrevivência dos em- briões. Os embriões apresentaram o maior comprimento de raiz (4,5 cm) e da haste caulinar (2,1 cm) com carvão ativado na concentração de 0,5 g/L e 2,0 g/L, respectivamente.

Poliembrionia e freqüência de híbridos em Citrus spp.

Com o objetivo de estudar a relação entre o grau de poliembrionia e a freqüência de híbridos em citros, foram realizados cruzamentos controlados utilizando como parentais femininos as tangerinas 'Clementina' e 'Sunki' e os limões 'Cravo' e 'Volkameriano', que apresentam graus de poliembrionia nulo, baixo, baixo a moderado e moderado, respectivamente. Como parentais masculinos empregaram-se Poncirus trifoliata e híbridos desta, tendo em vista a característica morfológica dominante folhas trilobadas. Observaram-se nas tangerinas 'Clementina' e 'Sunki' e nos limões 'Cravo' e 'Volkameriano' as seguintes freqüências de híbridos: 57,4%, 53,6%, 18,6% e 14,2%, respectivamente. Foi verificada uma associação negativa entre o grau de poliembrionia e a freqüência de híbridos.

Freqüência de híbridos em cruzamento entre tangerina 'cravo' e laranja 'pêra': análise de marcadores morfológicos e RAPD

Os objetivos deste trabalho foram avaliar a freqüência de híbridos de cruzamento entre tangerina 'Cravo' (Citrus reticulata Blanco) e laranja 'Pêra' (Citrus sinensis (L.) Osbeck), o uso de marcadores morfológicos e moleculares (RAPD) na identificação precoce de plantas zigóticas, e a variabilidade dos híbridos. A porcentagem de híbridos foi maior na população germinada em placas de Petri (19,4%). Verificou-se que quanto maior a competição entre os "seedlings" por espaço e nutrientes, menor a freqüência de plantas híbridas. A identificação dos híbridos não foi possível apenas com o uso de marcadores morfológicos. A análise morfológica dos híbridos revelou elevada variabilidade.

Comparison of citrus rooting evaluation methods using root images in soil profiles and root weight

A digitized image method was compared with a standard washing technique for measuring citrus roots in the field. Video pictures of roots were taken in a soil profile. The profile area analyzed was defined by iron rings, which were also used to remove the roots to determine their dry weight. The roots presented in the pictures were quantified using SIARCS software developed by Embrapa. The root length and area determined by digital images provided a good estimate of root quantity present in the profile.

Agrobacterium-mediated genetic transformation of 'Hamlin' sweet orange

The development and optimization of efficient transformation protocols is essential in new citrus breeding programs, not only for rootstock, but also for scion improvement. Transgenic 'Hamlin' sweet orange (Citrus sinensis (L.) Osbeck) plants were obtained by Agrobacterium tumefaciens-mediated transformation of epicotyl segments collected from seedlings germinated in vitro. Factors influencing genetic transformation efficiency were evaluated including seedling incubation conditions, time of inoculation with Agrobacterium and co-culture conditions. Epicotyl segments were adequate explants for transformation, regenerating plants by direct organogenesis. Higher percentage of transformation was obtained with explants collected from seedlings germinated in darkness, transferred to 16 hours photoperiod for 2-3 weeks, and inoculated with Agrobacterium for 15-45 min. The best co-culture condition was the incubation of the explants in darkness, for three days in culture medium supplemented with 100 muM of acetosyringone. Genetic transformation was confirmed by performing beta-glucoronidase (GUS) assays and, subsequently, by PCR amplification for the nptII and GUS genes.

Characterization of mandarin citrus germplasm from Southern Brazil by morphological and molecular analyses

The objective of this work was to characterize mandarin (Citrus spp.) germplasm from Southern Brazil by morphological and molecular analyses. Thirty seven cultivars from 34 distinct mandarin varieties were evaluated by morphological and agronomic traits of leaves, flowers and fruits, and by microsatellite markers. The morphological and agronomic characteristics suggested that almost all varieties can be produced for commercial use, and some, as the Satsuma variety, are recommended for breeding programs. Pooled DNA samples from 1-5 plants belonging to each cultivar were tested. Eight of the nine primers detected polymorphisms. Specific markers were found for some accessions. The dendrogram constructed with the morphological results divided the 37 cultivars into four groups, while that obtained with the microsatellites clustered 35 of the 37 cultivars into three groups only. Generally, intervarietal differences are not high, and this lack of agreement in the two multifactorial analyses indicates that diverse evolutionary factors are acting at these two levels of investigation.

Selection processes in a citrus hybrid population using RAPD markers

The objective of this work was to evaluate the processes of selection in a citrus hybrid population using segregation analysis of RAPD markers. The segregation of 123 RAPD markers between 'Cravo' mandarin (Citrus reticulata Blanco) and 'Pêra' sweet orange (C. sinensis (L.) Osbeck) was analysed in a F1 progeny of 94 hybrids. Genetic composition, diversity, heterozygosity, differences in chromosomal structure and the presence of deleterious recessive genes are discussed based on the segregation ratios obtained. A high percentage of markers had a skeweness of the 1:1 expected segregation ratio in the F1 population. Many markers showed a 3:1 segregation ratio in both varieties and 1:3 in 'Pêra' sweet orange, probably due to directional selection processes. The distribution analysis of the frequencies of the segregant markers in a hybrid population is a simple method which allows a better understanding of the genetics of citrus group.

Stomatal analysis of citrus somatic hybrids obtained by protoplast fusion

The objective of this work was to evaluate leaf epidermis morphological characteristics of three citrus somatic hybrids, compared to their parents. Parental and somatic hybrid young leaves were collected and processed for scanning electron microscope observations. Citrus polyploid hybrids have fewer stomata per area and these are larger compared to their diploid parental parents. No differences in internal arrangement of the stomatal cells were detected between parental plants and somatic hybrids. Additional studies may determine if these differences will influence physiological behavior of the plants in the field.

Somatic hybridization between Citrus sinensis (L.) Osbeck and C. grandis (L.) Osbeck

This work had as objective to produce citrus somatic hybrids between sweet oranges and pummelos. After chemical fusion of sweet orange embryogenic protoplasts with pummelo mesophyll-derived protoplasts, plants were regenerated by somatic embryogenesis and acclimatized in a greenhouse. The hybrids of 'Hamlin' sweet orange + 'Indian Red' pummelo and 'Hamlin' sweet orange + 'Singapura' pummelo were confirmed by leaf morphology, chromosome counting and molecular analysis. These hybrids have potential to be used directly as rootstocks aiming blight, citrus tristeza virus, and Phytophthora-induced disease tolerance, as well as for rootstocks improvement programs.

Olfactory mediated interactions between Citrus aurantium Toxoptera citricida and Lysiphlebus testaceipes

The objective of this study was to establish whether there are olfactory interactions in the Lysiphlebus testaceipes Toxoptera citricida and Citrus aurantium tritrophic system. The response of male and female L. testaceipes to different odour sources of the host plant C. aurantium, the aphid host T. citricida and aphid-plant complex were investigated using a Y-tube olfactometer. Laboratory experiments were conducted by exposing individually aged male and female L. testaceipes to eight different odour treatments. Response of the parasitoids was taken after 15 min exposure to the volatiles from the different odour sources and based on their orientation to the particular chamber. Seventy percent of both male and female L. testaceipes showed high attractivity to aphid infested leaves. There was no significant difference based on age and sex of the parasitoid on their choice of odour. The organic compounds released by these combinations acted as semiochemicals in the tritrophic interactions and it is suggested that insect feeding induced attraction of the parasitoid L. testaceipes.