EFFECTS OF SILICON ON ALLEVIATING ARSENIC TOXICITY IN MAIZE PLANTS

Arsenic is a metalloid highly toxic to plants and animals, causing reduced plant growth and various health problems for humans and animals. Silicon, however, has excelled in alleviating stress caused by toxic elements in plants. The aim of this study was to investigate the effects of Si in alleviating As stress in maize plants grown in a nutrient solution and evaluate the potential of the spectral emission parameters and the red fluorescence (Fr) and far-red fluorescence (FFr) ratio obtained in analysis of chlorophyll fluorescence in determination of this interaction. An experiment was carried out in a nutrient solution containing a toxic rate of As (68 μmol L-1) and six increasing rates of Si (0, 0.25, 0.5, 1.0, 1.5, and 2.0 mmol L-1). Dry matter production and concentrations of As, Si, and photosynthetic pigments were then evaluated. Chlorophyll fluorescence was also measured throughout plant growth. Si has positive effects in alleviating As stress in maize plants, evidenced by the increase in photosynthetic pigments. Silicon application resulted in higher As levels in plant tissue; therefore, using Si for soil phytoremediation may be a promising choice. Chlorophyll fluorescence analysis proved to be a sensitive tool, and it can be successfully used in the study of the ameliorating effects of Si in plant protection, with the Fr/FFr ratio as the variable recommended for identification of temporal changes in plants.

Soil Aggregation, Organic Carbon Concentration, and Soil Bulk Density As Affected by Cover Crop Species in a No-Tillage System

Soil aggregation and the distribution of total organic carbon (TOC) may be affected by soil tillage and cover crops. The objective of this study was to determine the effects of crop rotation with cover crops on soil aggregation, TOC concentration in the soil aggregate fractions, and soil bulk density under a no-tillage system (NTS) and conventional tillage system (CTS, one plowing and two disking). This was a three-year study with cover crop/rice/cover crop/rice rotations in the Brazilian Cerrado. A randomized block experimental design with six treatments and three replications was used. The cover crops (treatments) were: fallow, Panicum maximum, Brachiaria ruziziensis, Brachiaria brizantha, and millet (Pennisetum glaucum). An additional treatment, fallow plus CTS, was included as a control. Soil samples were collected at the depths of 0.00-0.05 m, 0.05-0.10 m, and 0.10-0.20 m after the second rice harvest. The treatments under the NTS led to greater stability in the soil aggregates (ranging from 86.33 to 95.37 %) than fallow plus CTS (ranging from 74.62 to 85.94 %). Fallow plus CTS showed the highest number of aggregates smaller than 2 mm. The cover crops affected soil bulk density differently, and the millet treatment in the NTS had the lowest values. The cover crops without incorporation provided the greatest accumulation of TOC in the soil surface layers. The TOC concentration was positively correlated with the aggregate stability index in all layers and negatively correlated with bulk density in the 0.00-0.10 m layer.

LEAF TOTAL NITROGEN CONCENTRATION AS AN INDICATOR OF NITROGEN STATUS FOR PLANTLETS AND YOUNG PLANTS OF EUCALYPTUS CLONES

The use of leaf total nitrogen concentration as an indicator for nutritional diagnosis has some limitations. The objective of this study was to determine the reliability of total N concentration as an indicator of N status for eucalyptus clones, and to compare it with alternative indicators. A greenhouse experiment was carried out in a randomized complete block design in a 2 × 6 factorial arrangement with plantlets of two eucalyptus clones (140 days old) and six levels of N in the nutrient solution. In addition, a field experiment was carried out in a completely randomized design in a 2 × 2 × 2 × 3 factorial arrangement, consisting of two seasons, two regions, two young clones (approximately two years old), and three positions of crown leaf sampling. The field areas (regions) had contrasting soil physical and chemical properties, and their soil contents for total N, NH+4-N, and NO−3-N were determined in five soil layers, up to a depth of 1.0 m. We evaluated the following indicators of plant N status in roots and leaves: contents of total N, NH+4-N, NO−3-N, and chlorophyll; N/P ratio; and chlorophyll meter readings on the leaves. Ammonium (root) and NO−3-N (root and leaf) efficiently predicted N requirements for eucalyptus plantlets in the greenhouse. Similarly, leaf N/P, chlorophyll values, and chlorophyll meter readings provided good results in the greenhouse. However, leaf N/P did not reflect the soil N status, and the use of the chlorophyll meter could not be generalized for different genotypes. Leaf total N concentration is not an ideal indicator, but it and the chlorophyll levels best represent the soil N status for young eucalyptus clones under field conditions.

Occurrence and Structure of Arbuscular Mycorrhizal Fungal Communities in Cassava after Cultivation of Cover Crops as Observed by the “PCR-DGGE” Technique

ABSTRACT Cassava (Manihot esculenta Crantz) is a highly mycotrophic crop, and prior soil cover may affect the density of arbuscular mycorrhizal fungi (AMFs), as well as the composition of the AMFs community in the soil. The aim of this study was to evaluate the occurrence and the structure of AMFs communities in cassava grown after different cover crops, and the effect of the cover crop on mineral nutrition and cassava yield under an organic farming system. The occurrence and structure of the AMFs community was evaluated through polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE). A randomized block experimental design was used with four replications. Six different cover crop management systems before cassava were evaluated: black oats, vetch, oilseed radish, intercropped oats + vetch, intercropped oats + vetch + oilseed radish, plus a control (fallow) treatment mowed every 15 days. Oats as a single crop or oats intercropped with vetch or with oilseed radish increased AMFs inoculum potential in soil with a low number of propagules, thus benefiting mycorrhizal colonization of cassava root. The treatments did not affect the structure of AMFs communities in the soil since the AMFs communities were similar in cassava roots in succession to different cover crops. AMFs colonization was high despite high P availability in the soil. The cassava crop yield was above the regional average, and P levels in the leaves were adequate, regardless of which cover crop treatments were used. One cover crop cycle prior to the cassava crop was not enough to observe a significant response in variables, P in plant tissue, crop yield, and occurrence and structure of AMFs communities in the soil. In the cassava roots in succession, the plant developmental stage affected the groupings of the structure of the AMF community.

Changes in Soil Organic Carbon Fractions in Response to Cover Crops in an Orange Orchard

ABSTRACT The cultivation of cover crops intercropped with fruit trees is an alternative to maintain mulch cover between plant rows and increase soil organic carbon (C) stocks. The objective of this study was to evaluate changes in soil total organic C content and labile organic matter fractions in response to cover crop cultivation in an orange orchard. The experiment was performed in the state of Bahia, in a citrus orchard with cultivar ‘Pera’ orange (Citrus sinensis) at a spacing of 6 × 4 m. A randomized complete block design with three replications was used. The following species were used as cover crops: Brachiaria (Brachiaria decumbes) – BRAQ, pearl millet (Pennisetum glaucum) – MIL, jack bean (Canavalia ensiformis) – JB, blend (50 % each) of jack bean + millet (JB/MIL), and spontaneous vegetation (SPV). The cover crops were broadcast-seeded between the rows of orange trees and mechanically mowed after flowering. Soil sampling at depths of 0.00-0.10, 0.10-0.20, and 0.20-0.40 m was performed in small soil trenches. The total soil organic C (SOC) content, light fraction (LF), and the particulate organic C (POC), and oxidizable organic C fractions were estimated. Total soil organic C content was not significantly changed by the cover crops, indicating low sensitivity in reacting to recent changes in soil organic matter due to management practices. Grasses enabled a greater accumulation of SOC stocks in 0.00-0.40 m compared to all other treatments. Jack bean cultivation increased LF and the most labile oxidizable organic C fraction (F1) in the soil surface and the deepest layer tested. Cover crop cultivation increased labile C in the 0.00-0.10 m layer, which can enhance soil microbial activity and nutrient absorption by the citrus trees. The fractions LF and F1 may be suitable indicators for monitoring changes in soil organic matter content due to changes in soil management practices.

Cover Crops and Nitrogen Fertilization Effects on Nitrogen Soil Fractions under Corn Cultivation in a No-Tillage System

ABSTRACT The use of cover crops has recently increased and represents an essential practice for the sustainability of no-tillage systems in the Cerrado region. However, there is little information on the effects of nitrogen fertilization and cover crop use on nitrogen soil fractions. This study assessed changes in the N forms in soil cropped to cover crops prior to corn growing. The experiment consisted of a randomized complete block design arranged in split-plots with three replications. Cover crops were tested in the plots, and the N topdressing fertilization was assessed in the subplots. The following cover species were planted in succession to corn for eight years: Urochloa ruziziensis, Canavalia brasiliensis M. ex Benth, Cajanus cajan (L.) Millsp, and Sorghum bicolor (L.) Moench. After corn harvesting, the soil was sampled at depths of 0.00-0.10 and 0.10-0.20 m. The cover crops showed different effects at different soil depths. The soil cultivated with U. ruziziensis showed higher contents of total-N and particulate-N than the soil cultivated with C. cajan. Particulate-N was the most sensitive to changes in the soil management among the fractions of N assessed. The soil under N topdressing showed a lower content of available-N in the 0.10-0.20 m layer, which may be caused by the season in which the sampling was conducted or the greater uptake of the available-N by corn.

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.

Effects of soil water depletion on the water relations in tropical kudzu

Tropical kudzu (Pueraria phaseoloides (Roxb.) Benth., Leguminosae: Faboideae) is native to the humid Southeastern Asia. Tropical kudzu has potential as a cover crop in regions subjected to dryness. The objective of this paper was to evaluate the effect of soil water depletion on leaflet relative water content (RWC), stomatal conductance (g) and temperature (T L) in tropical kudzu. RWC of waterstressed plants dropped from 96 to 78%, following a reduction in SWC from 0.25 to 0.17 g (H2O).g (dry soil)-1.Stomatal conductance of stressed plants decreased from 221 to 98 mmol.m-2.s-1, following the reduction in soil water content (SWC). The day after re-irrigation, g of water stressed plants was 15% lower than g of unstressed plants. Differences in T L between waterstressed and unstressed plants (deltaT L) rose linearly from 0.1 to 2.2ºC following progressive water deficit. RWC and T L of waterstressed plants paralled RWC and T L of unstressed plants the day after reirrigation. The strong decrease in SWC found in this study only induced moderate water stress in tropical kudzu. In addition, tropical kudzu recover rapidly from the induced water stress after the re-irrigation.

Aspects of leaf anatomy of kudzu (Pueraria lobata, Leguminosae-Faboideae) related to water and energy balance

Kudzu is a cover crop that has escaped cultivation in some subtropical and warm temperate regions. Kudzu has previously demonstrated broad intraspecific physiological plasticity while colonizing new environments. The objective of this paper was to investigate characteristics of kudzu leaflet anatomy that might contribute to its successful growth in climatically distinct environments, and to escape cultivation as well. Fresh and fixed leaflet strips of field-grown plants were analyzed. The lower epidermis of kudzu showed a higher frequency of stomata (147 ± 19 stomata mm-2) than the upper epidermis (26 ± 17 stomata mm-2). The average number of trichomes per square milimeter was 8 for both the upper and the lower epidermis. The average trichome length was 410 ± 200 mum for the upper epidermis and 460 ± 190 mum for the lower epidermis. Cuticle thickness was not considerably different between lower and upper epidermis. The leaflet blade consisted basically of two layers (upper and lower) of unicellular epidermis, two layers of palisade parenchyma and one layer of spongy parenchyma. One layer of paraveinal mesophyll was found between palisade and spongy parenchyma. In conclusion, leaflets of kudzu present anatomical characteristics that might contribute to the broad physiological plasticity shown by kudzu.

Albedo and estimates of net radiation for green beans under polyethylene cover and field conditions

This paper describes the albedo (r) and estimates of net radiation and global solar irradiance for green beans crop (Phaseolus vulgaris L.), cultivated in greenhouse with cover of polyethylene and field conditions, in Botucatu, SP, Brazil (22º 54' S; 48º 27' W; 850 m). The solar global irradiance (Rg) and solar reflected radiation (Rr) were used to estimate the albedo through the ratio between Rr and Rg. The diurnal curves of albedo were obtained for days with clear sky and partially cloudy conditions, for different phenological stages of the crop. The albedo ranged with the solar elevation, the environment and the phenological stages. The cloudiness range have almost no influence on the albedo diurnal amount. The estimation of radiation were made by linear regression, using the global solar irradiance (Rg) and net short-waves radiation (Rc) as independent variables. All estimates of radiation showed better adjustment for specific phenological periods compared to the entire crop growing cycle. The net radiation in the greenhouse has been estimated by the global solar irradiance measured at field conditions.

Effect of intercropping wheat with forage legumes on wheat production and ground cover

The use of winter legumes in southern Brazil is hindered by the slow growth of these species during establishment exposing soil surface to erosion. Introduction of these species along with spring wheat (Triticum aestivum L.) was studied as a means of increasing ground cover during their initial establishment period, without reducing wheat grain yield. Two experiments were conducted in nearby areas, one in each year. Birdsfoot trefoil (Lotus corniculatus L.), red clover (Trifolium pratense L.) cultivar Quiñequelli, white clover (T. repens L.), and arrowleaf clover (T. vesiculosum Savi) did not reduce cereal yield in either year. Wheat yield was reduced by intercropped red clover cultivar Kenland and by subclover (T. subterraneum L.) in the first year. No grain yield differences due to intercropping with any legume were detected in the second year, when rainfall was below normal. Intercropping with wheat showed to be a practical alternative to enhance ground cover at establishing forage legumes.

Critical values of nitrogen indices in tomato plants grown in soil and nutrient solution determined by different statistical procedures

The objective of this study was to establish critical values of the N indices, namely soil-plant analysis development (SPAD), petiole sap N-NO3 and organic N in the tomato leaf adjacent to the first cluster (LAC), under soil and nutrient solution conditions, determined by different statistical approaches. Two experiments were conducted in randomized complete block design with four repli-cations. Tomato plants were grown in soil, in 3 L pot, with five N rates (0, 100, 200, 400 and 800 mg kg-1) and in solution at N rates of 0, 4, 8, 12 and 16 mmol L-1. Experiments in nutrient solution and soil were finished at thirty seven and forty two days after transplanting, respectively. At those times, SPAD index and petiole sap N-NO3 were evaluated in the LAC. Then, plants were harvested, separated in leaves and stem, dried at 70ºC, ground and weighted. The organic N was determined in LAC dry matter. Three statistical procedures were used to calculate critical N values. There were accentuated discrepancies for critical values of N indices obtained with plants grown in soil and nutrient solution as well as for different statistical procedures. Critical values of nitrogen indices at all situations are presented.

A simple and reliable method for the screening of transgenic tobacco plants

Even though much improvement has been made in plant transformation methods, the screening of transgenic plants is often a laborious work. Most approaches for detecting the transgene in transformed plants are still timeconsuming, and can be quite expensive. The objective of this study was to search for a simpler method to screen for transgenic plants. The infiltration of kanamycin (100 mg/mL) into tobacco leaves resulted in conspicuous chlorotic spots on the non-transgenic plant leaves, while no spots were seen on the leaves of transformed plants. This reaction occurred regardless of age of the tested plants, and the method has proven to be simple, fast, non-destructive, relatively cheap, and reliable. These results were comparable to those obtained by the polymerase chain reaction (PCR) amplification of the transgene using specific primers.

Genetic variability of Brazilian phytoplasma and spiroplasma isolated from maize plants

The objective of this work was to characterize the genetic variability of phytoplasma and Spiroplasma kunkelii isolated from maize plants showing symptoms of stunt collected from different Brazilian geographic regions. A DNA fragment of 500 base pairs (bp) was amplified from the spiralin gene in S. kunkelii and one fragment of 1,200 bp was generated from 16S rDNA gene in phytoplasma. The partial sequences of the spiralin gene showed similarity of 98% among the isolates of S. kunkelii analyzed. These sequences were compared with the sequence of the spiralin gene from other Spiroplasma species deposited in the GenBank, resulting in a similarity varying from 76.9% to 88.1%. The 16S rDNA sequence from the phytoplasma were completely similar within the Brazilian isolates and showed up to 98% of the similarity with sequences already found from other phytoplasmas. A very narrow genetic variability was detected by these gene fragments within phytoplasma and Spiroplasma analyzed. However, other genomic regions with higher polymorphic levels shall be identified in order to better evaluate the genetic diversity within these microorganisms population.

Rescue of a non-viable accession and rapd analysis of recovered plants of Arachis retusa

In vitro regeneration of Arachis retusa was examined for the purpose of germplasm renewal and conservation. Random amplified polymorphic DNA (RAPD) fingerprinting was used to evaluate the genetic stability of plants derived from embryo axes and apical segments. Ten arbitrary decamer primers were screened and five of them were selected. Ninety genomic regions were evaluated, with an average of 18 loci per clone. All amplified segments were monomorphic. The results indicate that recovered plants are genetically stable at the assessed genomic regions and that both regeneration processes are suitable for in vitro germplasm preservation of Arachis species.