Atlantic rainforest natural regeneration in fragmented formations affected by increasing human disturbance.

Forests provides major ecosystem services worldwide. The Brazilian Atlantic Forest (BAF) has been dramatically devastated, with fragmentation processes jeopardizing its long-term sustainability. This study investigated the structure and successional dynamics in BAF natural regeneration along an anthroposequence characterized by increasing human disturbance histories as: secondary (SF) > disturbed (DF) > late forest (LF). We aimed to understand how and the degree to which BAF fragmentation and human disturbance affected plants, soils, and the whole soil-plant relationships and feedbacks. We investigated the natural regeneration conditions of plants (using plant classification and quali-quantitative analyses) and soil chemistry (including pH-CaCl2, H + Al, C, N, Pt, cation-exchange capacity (CEC), exchangeable cations, Al, B, Cu, Fe, K, Mn, and Zn) at twelve permanent, 2000 m2 plots, distributed across LF, DF, and SF forests. Significant differences were determined by ANOVA. Correlation matrix (CM) and factor analysis (FA) were used for understanding correlations and feedbacks/variability among investigated parameters, respectively. Most of investigated plant and soil parameters showed significant differences (p < 0.05) between more developed plant formations (LF) vs less developed ones (SF), with differences mainly due to soil's development stage. All investigated forest formations are featured by a great influence of the soil-plant relationships and feedbacks, with a decreasing magnitude as LF → DF → SF. Thus, there is a direct, statistically recognizable impact of both "recent" as well as "ancient" human disturbance on investigated soil-plant formations. The anthropogenic influence clearly affected not only plant and soil as "separate" systems but the whole complex of interactions and feedbacks among ecosystem components. A decreasing quality in soil and plant parameters was observed as human disturbance increased. We demonstrated that BAF plant and soil require decades for their recovery after human disturbances, with complex mechanisms and behaviors in the relationships among ecosystem components. The results can be useful for managing future recovery in an ecosystem of worldwide strategic importance.

Survival of thermotolerant coliforms in municipal biosolids after application in tropical soil cultivated with Eucalyptus.

The increasing production of biosolids in urban areas has been led to a search for alternative disposal avenues of this residue, which is rich in organic matter and nutrients. Agricultural land application of biosolids, motivated by its fertilizing power, is a widespread practice in many countries, but there are safety and regulatory concerns regarding the presence of pathogens in Class B biosolids. In addition, the scarcity of studies under tropical climate conditions raises questions that impede the agricultural use of this residue in some regions. The objective of this study was to evaluate the survival of thermotolerant coliforms over 12 months (52 weeks) after biosolids application on the surface of a Quartzipsamment neosol cultivated with Eucalyptus. Two different biosolids were studied: those generated by a biological treatment system with complete mixing aeration ponds followed by decantation ponds (Treatment A) and biosolids from a biological treatment system with conventional activated biosolids reactors (Treatment B), both delineated in randomized blocks with four replicates. After application on the forest soil, we estimated an average survival time of 54 weeks for thermotolerant coliforms present in Treatment A biosolids and 93 weeks in Treatment B biosolids. Thermotolerant coliforms persist much longer under tropical climate conditions in Brazil than in comparable studies under temperate climate conditions. This reaffirms the need to carry out studies covering the full range of moisture and temperature regimes in which biosolids are applied as fertilizer.

Evaluating carbon stocks in soils of fragmented Brazilian Atlantic Forests (BAF) based on soil features and different methodologies.

Brazil's Atlantic Forest (BAF) is a highly fragmented, strategic environmental and socio-economic region that represents the fourth biodiversity hotspot while also producing many commodities that are exported globally. Human disturbance plays a pivotal role as a driver of BAF's soil dynamics and behaviors. The soils under Late Primary and Secondary Semideciduous Seasonal Forests (LPSF and LSSF) were characterized by high to moderate resilience, with improved chemical properties as human disturbance decreased. The Transitional Forest to Cerrado (TFC) had the worst soil conditions. Disturbed Primary and Secondary Semideciduous Seasonal Forests (DPSF and DSSF) represent a transitional stage between LPSF/LSSF and TFC. Accordingly, SOCs stocks increased from TFC << DPSF, DSSF < LPSF, LSSF. In BAF soils, to avoid unreliable data, SOCs measurements should be (i) conducted to at least 1 m soil depth and (ii) quantified with a CHN analyzer. Human disturbance strongly affected the positive feedback between vegetation succession, SOCs, and soil nutrition. Soil development decreased as human disturbance increased, thus negatively affecting SOCs. Soils in the BAF require a long time to recover after the end of human disturbance, thus suggesting that preservation strategies should be prioritized in remnant BAF fragments.

Successive sewage sludge fertilization: Recycling for sustainable agriculture.

Sewage sludge (SS) is widely used in agriculture in several countries around the world. However, the impact of successive applications of SS on soil and the risks of nutrient leaching are often neglected. In this study, corn was grown on a constructed wetland for four crop cycles (two years), in which the wetland was subjected to successive SS applications. The objective of this study was to evaluate how the successive applications of SS affect the availability and leaching of nutrients in the soil profile, after two years of cultivation. Experiments were performed using a randomized block design with repeated measurements in time, that is, soil was sampled in each harvest. Six treatments were tested: four fertilizations based on sewage sludge, resulting from biological and anaerobic treatment, calculated to provide 25 (SS25), 50 (SS50), 75 (SS75), and 100% (SS100), of the N required for corn production (140 kg ha-1); a mineral fertilization (NPK) (140 kg ha-1 N, 70 kg ha-1 of P2O5 and 40 kg ha-1 of K2O) and a control (without fertilization). The results showed that four consecutive applications of SS100 for two years caused significant accumulation of nutrients and organic matter in the superficial layers of the soil. The electrical conductivity and the concentration of NO3- in the soil solution were higher than those permitted by Brazilian legislation. The adoption of domestic SS in Brazilian agriculture can be a viable alternative in the search for an environment-friendly and economically feasible method for SS disposal.

Current status, uncertainty and future needs in soil organic carbon monitoring.

Increasing human demands on soil-derived ecosystem services requires reliable data on global soil resources for sustainable development. The soil organic carbon (SOC) pool is a key indicator of soil quality as it affects essential biological, chemical and physical soil functions such as nutrient cycling, pesticide and water retention, and soil structure maintenance. However, information on the SOC pool, and its temporal and spatial dynamics is unbalanced. Even in well-studied regions with a pronounced interest in environmental issues information on soil carbon (C) is inconsistent. Several activities for the compilation of global soil C data are under way. However, different approaches for soil sampling and chemical analyses make even regional comparisons highly uncertain. Often, the procedures used so far have not allowed the reliable estimation of the total SOC pool, partly because the available knowledge is focused on not clearly defined upper soil horizons and the contribution of subsoil to SOC stocks has been less considered. Even more difficult is quantifying SOC pool changes over time. SOC consists of variable amounts of labile and recalcitrant molecules of plant, and microbial and animal origin that are often operationally defined. A comprehensively active soil expert community needs to agree on protocols of soil surveying and lab procedures towards reliable SOC pool estimates. Already established long-term ecological research sites, where SOC changes are quantified and the underlying mechanisms are investigated, are potentially the backbones for regional, national, and international SOC monitoring programs.

Eucalyptus ESTs related to genes for oxidative stress.

Oxidative stress generating active oxygen species has been proved to be one of the underlying agents causing tissue injury after the exposure of Eucalyptus (Eucalyptus spp.) plants to a wide variety of stress conditions. The objective of this study was to perform data mining to identify favorable genes and alleles associated with the enzyme systems superoxide dismutase, catalase, peroxidases, and glutathione S-transferase that are related to tolerance for environmental stresses and damage caused by pests, diseases, herbicides, and by weeds themselves. This was undertaken by using the eucalyptus expressed-sequence database (https//forests.esalq.usp.br). The alignment results between amino acid and nucleotide sequences indicated that the studied enzymes were adequately represented in the ESTs database of the FORESTs project.

Boron transport in Eucalyptus. 2. Identification in silico of a putative boron transporter for xylem loading in eucalypt

Boron (B) is a low mobility plant micronutrient whose molecular mechanisms of absorption and translocation are still controversial. Many factors are involved in tolerance to Boron excess or deficiency. Recently, the first protein linked to boron transport in biological systems, BOR1, was characterized in Arabidopsis thaliana. This protein is involved in boron xylem loading and is similar to bicarbonate transporters found in animals. There are indications that BOR1 is a member of a conserved protein family in plants. In this work, FORESTS database was used to identify sequences similar to this protein family, looking for a probable BOR1 homolog in eucalypt. We found five consensus sequences similar to BOR1; three of them were then used in multiple alignment analysis. Based on amino acid similarity and in silico expression patterns, a consensus sequence was identified as a candidate BOR1 homolog, helping deeper experimental assays that could identify the function of this protein family in Eucalyptus

Eucalyptus ESTs associated with resistance to herbicide inhibitors of aromatic and branched-chain amino acid synthesis

Herbicides inhibit enzymatic systems of plants. Acetolactate synthase (ALS, EC = 4.1.3.18) and 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS, EC 2.5.1.19) are key enzymes for herbicide action. Hundreds of compounds inhibit ALS. This enzyme is highly variable, enabling the selective control of weeds in a number of crops. Glyphosate, the only commercial herbicide inhibiting EPSPS is widely used for non-selective control of weeds in many crops. Recently, transgenic crops resistant to glyphosate were developed and have been used by farmers. The aim of this study was the data mining of eucalypt expressed sequence tags (ESTs) in the FORESTs Genome Project database (https://forests.esalq.usp.br) related to these enzymes. Representative amino acid sequences from the NCBI database associated with ALS and EPSPS were blasted with ESTs from the FORESTs database using the tBLASTx option of the blast tool. The best blasting reads and clusters from FORESTs, represented as nucleotide sequences, were blasted back with the NCBI database to evaluate the level of similarity with available sequences from different species. One and seven clusters were identified as showing high similarity with EPSPS and ALS sequences from the literature, respectively. The alignment of EPSPS sequences allowed the identification of conserved regions that can be used to design specific primers for additional sequencings

Eucalyptus ESTs corresponding to the enzyme glutamine synthetase and the protein D1, sites of action of herbicides that cause oxidative stress

This work was aimed at locating Eucalyptus ESTs corresponding to the GS enzyme (Glutamine Synthetase, EC = 6.3.1.2) and to the D1 protein, which are directly related to resistance to herbicides that promote oxidative stress. Glutamine Synthetase corresponds to the site of action of the herbicide glufosinate. Herbicides that belong to groups such as ureas, uracils, triazines and triazinones act on the D1-Qb complex (receptor of electrons from the Photosystem II) by inactivating it. The clusters EGEQRT3302E01.g, EGEQRT3001F12.b; EGEZLV1203B04.g; EGBGFB1211H06.g and EGEZLV1205F09.g enclosed complete sequences (with 356 amino acids) of the Glutamine Synthetase enzyme. The cluster EGEQSL1054G06.g is a consensus of four reads and enclosed a complete sequence of D1 Protein (with 353 amino acids). The comparison of the sequences of Protein D1 from different species showed that the substitutions of serine (S) by glycine (G) or serine (S) by threonine (T) at the position 264 could produce plants resistant to herbicides that act on electron flow on Photosystem II. The sequence of amino acids corresponding to the cluster EGEQSL1054G06.g had a serine in position 264 indicating sensitivity of the Eucalyptus plants to herbicides that act on this site

Eucalyptus ESTs corresponding to the protoporphyrinogen IX oxidase enzyme related to the synthesis of heme, chlorophyll, and to the action of herbicides

This work was aimed at locating Eucalyptus ESTs corresponding to the PROTOX or PPO enzyme (Protoporphyrinogen IX oxidase, E.C. 1.3.3.4) directly related to resistance to herbicides that promote oxidative stress, changing the functionality of this enzyme. PROTOX, which is the site of action of diphenyl-ether (oxyfluorfen, lactofen, fomesafen), oxadiazole (oxadiazon and oxadiargyl), and aryl triazolinone (sulfentrazone and carfentrazone) herbicides, acts on the synthesis route of porphyrins which is associated with the production of chlorophyll a, catalases, and peroxidases. One cluster and one single read were located, with e-values better than e-70, associated to PROTOX. The alignment results between amino acid sequences indicated that this enzyme is adequately represented in the ESTs database of the FORESTs project