Print-capture PCR for detection of tomato begomoviruses from plants and whiteflies

Print-capture (PC) Polymerase chain reaction (PCR) was evaluated as a novel detection method of plant viruses. Tomato (Lycopersicon esculentum) plants infected with begomovirus (fam. Geminiviridae, gen. Begomovirus) and viruliferous whiteflies were used to study the efficiency of the method. Print-capturing steps were carried out using non-charged nylon membrane or filter paper as the solid support for DNA printings. Amplified DNA fragments of expected size were consistently obtained by PCR from infected plants grown in a greenhouse, after direct application of printed materials to the PCR mix. However, virus detection from a single whitefly and from field-grown tomato samples required a high temperature treatment of printed material prior to PCR amplification. Comparison of nylon membrane and filter paper as the solid support revealed the higher efficiency of the nylon membrane. The application of print-capture PCR reduces the chances of false-positive amplification by reducing manipulation steps during preparation of the target DNA. This method maintains all the advantages of PCR diagnosis, such as the high sensitivity and no requirement of radioactive reagents.

Etiology of phomopsis root rot in soybean

In a survey of damages caused by soybean root rot to crops in the south of Brazil for several years, a root rot caused by Phomopsis sp has been found with increasing frequency. The primary symptoms are seen when the main root is cut longitudinally, including the death of the wood which shows white coloration and well-defined black lines that do not have a defined format. Thus, based on similarity, it has been called geographic root rot due to its aspect resembling irregular lines that separate regions on a map. In isolations, colonies and alpha spores of Phomopsis have prevailed. Pathogenicity test was done by means of inoculation in the crown of plants cultivated in a growth chamber. The geographic symptoms were reproduced in plants and the fungus Phomopsis sp. was reisolated. In soybean stems naturally infected with pod and stem blight, geographic symptoms caused by Phomopsis phaseoli are found. To the known symptoms on stems, pods and grains, that of root rot caused by P. phaseoli is now added.

Outplanting performace of eucalyptus clonal cuttings produced in different containers and substrates

The objective of this work was to evaluate the outplanting growth of Eucalyptus grandis e E. saligna clones, produced by cuttings in tubes (50cm³) and in pressed blocks (40x60x07cm) - 175 cm³/ seedlings, with different substrates (BT - sugarcane bagasse+sugarcane filter cake; AR - carbonized rice hull + eucalyptus bark; TF - peat). The experiment was arranged in a randomized block design, in a 2x7 factorial (2 clones and 7 treatments), with four replicates with 25 plants. Survival was evaluated two months later. Plant growth was monitored through height and ground level diameter at 20, 40, 60, 120 and 180 days after outplanting. To evaluate the effect of the containers on stem and root biomass in both clones, 180 days after outplanting, the cuttings grown in BT substrate with fertilizer were selected. One plant per plot of each clone, grown in tubes and in pressed blocks was selected. The E. grandis and E. saligna cuttings grown in pressed blocks with sugarcane bagasse+sugarcane filter cake presented greater height and diameter after out planting. Both clones presented larger root, bark, log and branch biomass production in plants produced in the block system. Cuttings of E. saligna grown in pressed blocks showed 80% most wood biomass 180 days after outplanting, compared to that grown in tubes. In E. grandis, the differences in diameter and height, in function of the cutting production system, decreased along time, while in E. saligna these differences increased along the evaluation period.

Development of experimental structure and influence of high CO2 concentration in maize cro

Maize is a C4 plant that shows few or no response to high [CO2]. Thus, this study aimed to analyze the photosynthetic rate and yield of maize under high [CO2] and develop open-top chambers (OTC) to create an atmosphere enriched with CO2. The experiment was conducted between October 2008 and March 2009. The OTCs were developed in modular scheme. Measurement of photosynthetic rates, transpiration, stomata conductance, grain yield and dry matter were performed. The experimental design was randomized blocks with four replications and three treatments: P1 - plants grown in OTC with 700 ppm [CO2], P2 - plants grown in OTC with environmental [CO2], and P3 - control, cultivated in open field. The results were analyzed by ANOVA and Tukey's test (Pr< 0.05). The chambers can reduce by 25% the photosynthetically active radiation and increase the air and leaf temperatures. Plants under high [CO2] (P1) showed the highest photosynthetic rates and the lowest stomata conductance and transpiration. The total weight of grains (g) and dry mass of shoots (g) showed no increases for P1, despite their higher photosynthetic rates.

Experimental swainsonine poisoning in goats ingesting Ipomoea sericophylla and Ipomoea riedelii (Convolvulaceae)

Ipomoea sericophylla and Ipomoea riedelii cause a glycoprotein storage disease in goats. This paper reports the experimental poisoning in goats by dried I. sericophylla and I. riedelii containing 0.05% and 0.01% swainsonine, respectively. Three groups with four animals each were used. Group 1 received daily doses of 2g/kg body weight (bw) of dried I. sericophylla (150mg of swainsonine/kg). Goats from this group had clinical signs 36-38 days after the start of ingestion. Group 2 received dried I. riedelii daily doses of 2g/kg of I. riedelii (30mg of swainsonine/kg) for 70 days. No clinical signs were observed, therefore the swainsonine dose was increased to 60mg/kg for another 70 days. Goats from Group 2 had clinical signs 26-65 days after increase in swainsonine dose to 60mg/kg. Group 3 was used as control. In these experiments the minimum toxic dose was 60mg/kg which represents 0.0004% of the dry matter in goats ingesting 1.5% bw of the dry matter. For goats ingesting 2%-2.5% bw of dry matter this dose would be 0.00024%-0.0003% of the dry matter. After the end of the experiment two goats were euthanized and another six were observed for recovery of clinical signs. Four goats that continued to consume swainsonine containing plant for 39-89 days after the first clinical signs had non reversible signs, while two goats that ingested the plant for only 15 and 20 days after the first clinical signs recovered completely. These and previous results indicate that irreversible lesions due to neuronal loss occur in goats that continue to ingest the plants for about 30 days after the first clinical signs. Clinical signs and histological lesions were similar to those reported previously for goats poisoned by swainsonine containing plants. No significant alterations were found in packed cell volume, red and white blood cell counts, hemoglobin and mean corpuscular hemoglobin concentrations, mean corpuscular volume, and serum levels of glucose, total protein, and albumin, and the serum activities of gamma glutamyl transferase and aspartate aminotransferase. Swainsonine concentration of 0.05% in I. sericophylla and 0.01% in I. riedelii are different from samples of these plants used in previous experiments, which contained 0.14% and 0.5% swainsonine, respectively, demonstrating a wide variation in the toxicity of different samples.

Persistence of the biocide activity of atrazine in soils of the southeast of Buenos Aires Province

Atrazine persistence in soils of the southeast of Buenos Aires Province, was studied by an oat bioassay. Atrazine doses of 0.58, 1.16, and 2.32 mg.g-1 dry soil weight (DSW) were applied to pots containing soils from Balcaree, A. Gonzáles Chaves and San Cayetano sites, whose organic matter (OM) content of soils were 5.70, 5.15, and 3,84%, respectively. Avena sativa cv. Millauquén plants were grownth in the pots under greenhouse conditions at different times after atrazine application. Shoots were evenly cut above the soil and dry weight determined as a measure of plant growth. Plants grown in non-sprayed soil were used as controls. Relative dry weight (RDW) of shoots was calculated as percentage of control. Atrazine phytotoxicity was expressed in terms of 50 % plant growth reduction (GR50) in the soils under study. Herbicide persistence was expressed in terms of days after treatment (DAT) needed for the plant to achieve 80% of RDW. Atrazine GR50 values of 0.30, 0.64, and 0.90 mg.g-1 DSW in soils from San Cayetano, Balcare and A.G. Chaves, were respectively obtained at 42 DAT. Herbicide persistences at the recommended dose (1.16 mg.g-1) were 100, 143, and 221 DAT for A.G. Chaves, Balcarce and San Cayetano soils, respectively. San Cayetano soil had both the lowest OM content and cation exchange capacity (CEC), as well as the highest pH, of all the soil studied here. These results were consistent with both the lowest GR50 and the highest persistence abtained for atrazine in this soil.

Growth, biomass allocation and photosynthesis of Rolandra fruticosa (asteraceae) in response to shade

The effects of shade on growth, biomass allocation patterns and photosynthetic response was examined for Rolandra fruticosa (L.) Kuntze, a common perennial weed shrub in cultivated pastures and agricultural areas of Brazilian Amazonia, for plants grown in full sunlight and those shaded to 30 % of full sunlight over a 34-d period. Specific leaf area and leaf area ratio were higher for shade plants during all the experimental period. Shade plants allocated significantly less biomass to root tissue than sun plants and relative growth rate was higher in sun plants. Sun leaves had significantly higher dark respiration and light saturated rates of photosynthesis than shade leaves. The apparent quantum efficiency was higher for shade leaves, while light compensation point was higher for sun leaves. These results are discussed in relation to their ecological and weed management implications.

Accumulation of dry mass and macronutrients by sourgrass plants

The experiment was carried out aiming to analyze the dry mass production and distribution and the content and accumulation of macronutrients in sourgrass (Digitaria insularis) plants cultivated under mineral nutrition standard conditions. Plants grew in 7-liter pots filled with sand substrate and daily irrigated with nutrient solution, being maintained under greenhouse conditions. Treatments consisted of times of evaluation (21, 35, 49, 63, 77, 91, 105, 119, and 133 days after emergence - DAE) and were arranged in a completely randomized design with four replicates. Sourgrass showed small accumulation of dry mass (0.3 g per plant) and macronutrients (3.7 mg of N per plant, 0.4 mg of P per plant, 5.6 mg of K per plant, 0.9 mg of Ca per plant, 0.7 mg of Mg per plant, and 0.3 mg of S per plant) at vegetative growth stage (< 49 DAE). Those accumulations increased mainly after 77 DAE, reaching the maximum theoretical value at 143, 135, 141, 129, 125, 120, and 128 DAE, for dry mass (12.4 g per plant), N (163.2 mg per plant), P (27.1 mg per plant), K (260.5 mg per plant), Ca (47.6 mg per plant), Mg (30.9 mg per plant), and S (13.7 mg per plant), respectively. K and N were found with higher rates and, as a consequence, they were required and accumulated in greater amounts in plant tissues of sourgrass.

Eucalyptus growth in silvopastoral system under different crown dia meters

The aim of this study was to evaluate the effect of different crown diameters on the early growth of eucalyptus intercropped with Brachiaria decumbens in a silvopastoral system. The experiment was conducted in a B. decumbens established pasture, where hybrid eucalyptus urograndis (clone GG100) was planted, spaced 8 x 3 m. A randomized block design was used, with six replicates. Treatments consisted of five crown diameters (0.0, 1.0, 1.5, 2.0, and 3.0 m) surrounding the eucalyptus plants. Five weeding hoes were performed throughout the experiment, according to the different crown diameters, aiming to maintain the eucalyptus plants free from B. decumbens interference. At 90, 180, 270, and 360 DAP, the height and the diameter of the eucalyptus plants were evaluated, and at 360 DAP, surface biomass and leaf area were evaluated. At 90 DAP, it was verified that the non-weeded plants had lower growth, compared to those submitted to crowns. Crown diameters of 2.51 and 2.64 m allowed greater growth in height and diameter at ground level of eucalyptus plants, respectively, in all periods evaluated. Biomass production and leaf area per plant at 360 DAP were also influenced by the different crown diameters. It was concluded that crown diameter around 2 meters provided favorable conditions for early growth of eucalyptus and less involvement in the area occupied by forage.

Aquatic plant community in porto primavera reservoir

Aiming to identify the populations of aquatic plants present in the Porto Primavera reservoir and evaluate the behavior of Hydrilla verticillata colonization of this water body a survey was carried out in 2007. The data was based on presence or absence, only were assessed the presence or absence of the species and the data were subjected to cluster analysis to establish differences in distribution and occurrence of populations. The community of aquatic plants showed 24 species distributed in 16 botanical families. Cyperaceae and Pontederiaceae were the most representative in terms of species richness. The submerged macrophyte Hydrilla verticillata showed the highest frequency of occurrence in the water body, showing a different behavior from the other populations of the water body. Species like Eichhornia crassipes, Eichhornia azurea, Typha dominguensis and Oxycaryum cubense also showed different behavior in relation to other populations within the community, forming large populations in lagoons and backwater areas.

MANAGEMENT OF MAIZE STOVER WITH UNDERDOSES RATES OF GLYPHOSATE APPLICATIONS IN NO-TILLAGE

ABSTRACTGlyphosate has significant effects on the growth and development of plants when in underdoses. This work was developed to verify the effect of the application of glyphosate in underdoses in lignin synthesis and consequently decomposition of maize stover. Two experiments were conducted; the first one in a greenhouse for underdoses adjustments and the second one in the production area. The experimental design of the first trial was completely randomized with four replications. The treatments consisted in the application of the underdoses: 0, 25, 50 and 100 g ha-1 of glyphosate. In the production area, the experimental design was a randomized block with four replications, in underdoses: 0, 12.5, 25 and 50 g ha-1. The results were submitted to analysis of variance and regression. The underdoses of 25 g a.e. ha-1in a greenhouse promoted 36% increase in productivity of stover, in addition to increasing the lignin content in 16%, with no change in the unwanted growth of maize plants. In the production area, the concentration of 12.5 g a.e. ha-1 of glyphosate reduced the lignin content and the other underdoses have not changed this feature in maize plants. None of the underdoses affected the height and biomass produced by the maize plants. The highest underdose tested promoted acceleration in the decomposition of maize stover.

Pollination and other biotic interactions in figs of Ficus eximia Schott (Moraceae)

During the period from 1992 to 1997, interactions of several organisms and Ficus eximia figs, a monoecious species, were studied in plants located in Campinas/SP and Londrina/PR (Brazil). Ficus eximia is pollinated by a single fig wasp species, Pegoscapus sp. (Hymenoptera: Agaonidae, Agaoninae), but also visited by other 14 non-pollinating wasps (Agaonidae, Eurytomidae, Torymidae). Mites (Tarsonemidae), nematodes (Diplogasteridae) and fungi which use the body of the pollinating fig wasp to disperse themselves were also observed.

Leaf optical properties of two liana species Canavalia parviflora Benth. and Gouania virgata Reissk in different light conditions

Lianas are plants that depend on support to reach some appreciable height, and they represent an important structural component of tropical forests. Although they predominate in clearings and gaps, some species survive in the understory. Changes in irradiance between these environments can affect leaf morphology and absorption of photosynthetic active radiation (PAR). We had examined the effects of different light regimes on leaf optical properties, chlorophyll content, specific leaf area, and leaf surface morphology in young seedlings of Canavalia parviflora Benth. (Fabaceae) and Gouania virgata Reissk (Rhamnaceae). The seedlings were distributed on workbenches covered by different layers of neutral shade netting, thus creating three levels of light intensity corresponding to about 40%, 10% and 1.5% of solar irradiance. Plants growing in full sun were used as a control. Both species exhibited an increase in reflectance in full sun and alterations in leaf morphology. Reduction in irradiance induced an increase in absorptance (decrease in reflectance and transmittance) in C. parviflora leaves in the green due to higher chlorophyll content. In G. virgata the spectral leaf changes were less observable. However, the efficiency of absorption was more pronounced in G. virgata than in C. parviflora leaves under 40%, 10% and 1.5% photon flux density (PFD). The greater efficiency of absorption in G. virgata was due to a larger specific leaf area (SLA) under these conditions. The adjustments in leaf optical properties can aid these species in overall carbon gain under limited light conditions.

Activity and functional interaction of alternative oxidase and uncoupling protein in mitochondria from tomato fruit

Cyanide-resistant alternative oxidase (AOX) is not limited to plant mitochondria and is widespread among several types of protists. The uncoupling protein (UCP) is much more widespread than previously believed, not only in tissues of higher animals but also in plants and in an amoeboid protozoan. The redox energy-dissipating pathway (AOX) and the proton electrochemical gradient energy-dissipating pathway (UCP) lead to the same final effect, i.e., a decrease in ATP synthesis and an increase in heat production. Studies with green tomato fruit mitochondria show that both proteins are present simultaneously in the membrane. This raises the question of a specific physiological role for each energy-dissipating system and of a possible functional connection between them (shared regulation). Linoleic acid, an abundant free fatty acid in plants which activates UCP, strongly inhibits cyanide-resistant respiration mediated by AOX. Moreover, studies of the evolution of AOX and UCP protein expression and of their activities during post-harvest ripening of tomato fruit show that AOX and plant UCP work sequentially: AOX activity decreases in early post-growing stages and UCP activity is decreased in late ripening stages. Electron partitioning between the alternative oxidase and the cytochrome pathway as well as H+ gradient partitioning between ATP synthase and UCP can be evaluated by the ADP/O method. This method facilitates description of the kinetics of energy-dissipating pathways and of ATP synthase when state 3 respiration is decreased by limitation of oxidizable substrate.

Plant-Herbivore Interaction in a Fragmented Landscape: Local Adaptation and Inbreeding

Reciprocal selection between interacting species is a major driver of biodiversity at both the genetic and the species level. This reciprocal selection, or coevolution, has led to the diversification of two highly diverse and abundant groups of organisms, flowering plants and their insect herbivores. In heterogeneous environments, the outcome of coevolved species interactions is influenced by the surrounding community and/or the abiotic environment. The process of adaptation allows species to adapt to their local conditions and to local populations of interacting species. However, adaptation can be disrupted or slowed down by an absence of genetic variation or by increased inbreeding, together with the following inbreeding depression, both of which are common in small and isolated populations that occur in fragmented environments. I studied the interaction between a long-lived plant Vincetoxicum hirundinaria and its specialist herbivore Abrostola asclepiadis in the southwestern archipelago of Finland. I focused on mutual local adaptation of plants and herbivores, which is a demonstration of reciprocal selection between species, a prerequisite for coevolution. I then proceeded to investigate the processes that could potentially hamper local adaptation, or species interaction in general, when the population size is small. I did this by examining how inbreeding of both plants and herbivores affects traits that are important for interaction, as well as among-population variation in the effects of inbreeding. In addition to bi-parental inbreeding, in plants inbreeding can arise from self-fertilization which has important implications for mating system evolution. I found that local adaptation of the plant to its herbivores varied among populations. Local adaptation of the herbivore varied among populations and years, being weaker in populations that were most connected. Inbreeding caused inbreeding depression in both plants and herbivores. In some populations inbreeding depression in herbivore biomass was stronger in herbivores feeding on inbred plants than in those feeding on outbred ones. For plants it was the other way around: inbreeding depression in anti-herbivore resistance decreased when the herbivores were inbred. Underlying some of the among-population variation in the effects of inbreeding is variation in plant phenolic compounds. However, variation in the modification of phenolic compounds in the digestive tract of the herbivore did not explain the inbreeding depression in herbivore biomass. Finally, adult herbivores had a preference for outbred host plants for egg deposition, and herbivore inbreeding had a positive effect on egg survival when the eggs were exposed to predators and parasitoids. These results suggest that plants and herbivores indeed exert reciprocal selection, as demonstrated by the significant local adaptation of V. hirundinaria and A. asclepiadis to one another. The most significant cause of disruption of the local adaptation of herbivore populations was population connectivity, and thus probably gene flow. In plants local adaptation tended to increase with increasing genetic variation. Whether or not inbreeding depression occurred varied according to the life-history stage of the herbivore and/or the plant trait in question. In addition, the effects of inbreeding strongly depended on the population. Taken together, inbreeding modified plant-herbivore interactions at several different levels, and can thus affect the strength of reciprocal selection between species. Thus inbreeding has the potential to affect the outcome of coevolution.