Use of semi-seletive media for detection of Sclerotinia sclerotiorum on bean and soybean seeds

This work was aimed at evaluating the possibility of using bromophenol blue as an indicator for detecting the presence of Sclerotinia sclerotiorum in the seeds of dry-beans (Phaseolus vulgaris) and soybean (Glycine max), through incubation of the seeds on an agar medium and "blotter" substrates. The seeds were artificially inoculated with four S. sclerotiorum isolates, plated on the agar medium, named Neon, and on modified Neon agar media all incubated at 14 and 20 ºC for seven days in the dark. Half of the seeds inoculated were surface desinfested prior to plating on the medium. The seeds showing change of colour in the medium, from blue to light yellow, as well as formation of typical mycelium and sclerotia in some cases, were considered to be infected or contaminated by S. sclerotiorum. The two incubation temperatures compared did not show significant (P<0.05) differences in detection level for most of the isolates tested on the different media. According to results obtained in this study, the Neon agar medium with incubation at 14 or 20 ºC has proved to be a reliable and quick method for the detection of S. sclerotiorum mycelium in naturally infected seeds of bean and soybean.

Effect of Fusarium graminearum and infection index on germination and vigor of maize seeds

Pathogens in maize (Zea mays) seeds cause serious problems, such as the loss of their capacity to germinative. The objectives of this study were to identify the optimal period for infection of maize seeds on agar colonized by Fusarium graminearum, when incubated for 4, 8, 16 and 32 h, and to evaluate the effect of the fungus on the germination and vigor of seeds with different infection levels. After the respective incubation periods, the seeds were removed from the culture medium and submitted to the blotter test for 3 min with and without superficial disinfection with 1% solution of sodium hypochlorite. Once the optimal period for seed incubation was identified, seeds from the same sample were again placed on the colonized agar for infection. Germination and vigor tests (accelerated aging and cold test) were performed with a mixture of healthy seeds (placed on PDA medium) and inoculated seeds, resulting in seeds with 0, 20, 40, 60, 80 and 100% rates of infection. The results showed that a period of 32 h was long enough to obtain seeds infected by the pathogen. There were no significant effects of fungal infection on seed germination at any of the infection levels, probably due to the high vigor of the maize seed lot tested. Regarding vigor tests, infection levels differed significantly from the control (0% infection), but there were no significant differences among the infection levels.

Detection of Rhynchosporium secalis in barley seeds from Argentina through polymerase chain reaction technique

Leaf scald of barley caused by Rhynchosporium secalis is an important disease in Argentina. The fungus is a necrotrophic pathogen which survives in stubble, seeds and weeds. Isolation of R. secalis from seeds on artificial media usually has not been successful due to the slow growth rate of the pathogen and strong inhibition by contaminants. The objective in this work was to detect R. secalis in different genotypes of barley seeds in Argentina using the polymerase chain reaction (PCR)-based diagnostic assay. Four barley genotypes were tested in 2004: Quilmes Ayelén, Quilmes Alfa, Barke and Maltería Pampa 1004. The previously described RS8 and RS9 primers were used for the detection of R. secalis in barley seeds. A 264-bp single band was obtained for each cultivar showing the presence of R. secalis. The use of specific primers was efficient in the detection of R. secalis in barley seeds in Argentina and could be used for routine diagnosis, epidemiology and seed transmission studies. This is the first report on the detection of R. secalis in barley seeds in Argentina.

Thermoanalytical study and characterization of native starches of Paraná pine seeds (Araucaria angustiofolia, Bert O. Ktze) and European chestnut seeds (Castanea sativa, Mill)

Starch is the most important carbohydrate storage in plants. It is a raw material with diverse botanical origins, and is used by the food, paper, chemical, pharmaceutical, textile and other industries. In this work, native starches of Paraná pine seeds (pinhão) (Araucária angustiofolia, Bert O. Ktze) and european chestnut seeds (Castanea sativa, Mill) were studied by thermoanalytical techniques: thermo-gravimetry (TG), differential thermal analysis (DTA) and differential scanning calorimetry (DSC), as well as X-ray powder patterns diffractometry. Apparent and total amylose content was also determined.

Detection, transmission, pathogenicity and chemical treatment of fungi in Ceiba speciosa seeds

The aim of this work was to evaluate fungus association, transmission and pathogenicity, besides chemical seed treatment in Ceiba speciosa seeds from different regions of southern Brazil. Seven seed samples were used to do the germination test, fungus detection by blotter test and potato-dextrose-agar (PDA), fungus transmission and pathogenicity tests; besides, chemical seed treatments were tested. Germination ranged from 0 to 59,5%. The following fungi were associated in the seeds: Fusarium sp., Alternaria sp., Colletotrichum sp., Curvularia sp. and Pestalotia sp.; in addition, Fusarium sp. was found in all the samples. Alternaria sp. and Fusarium sp. were transmitted by seeds. The isolates of Alternaria sp., Colletotrichum sp. and Fusarium sp., were pathogenic to seedlings and seeds. The seed treatment with methyl tiophanate and the combination captan + methyl tiophanate reduced Fusarium sp. incidence.

Fungicides, seed dresser adjuvants and storage time in the control of Drechslera teres in barley seeds

In experiments conducted in laboratory, the effect of fungicides, seed dresser adjuvants and storage time in the control of Drechslera teres in seeds of barley cultivar BRS Elis, with 58% incidence, was quantified. Fungicides indicated by barley research (carboxin + thiram, difenoconazole and triadimenol) compared with the mixture carbendazim + iprodione were tested. As seed dresser adjuvants, water (500m mL/100 Kg) and a polymer (150 mL/100 Kg) were used. Treated seeds were stored in paper bags and kept in the refrigerator at 5ºC. At 30-day intervals during six months, seeds were plated on semi-selective Reis's medium (1983). The best control was obtained by the mixtures carboxin + thiram and carbendazim + iprodione and the polymer as seed dresser. The control efficiency was improved by the storage time without negatively affecting seed germination. Due to the transmission efficiency, the fungus eradication in seeds should be pursued.

Semi-selective culture medium for Exserohilum turcicum isolation from corn seeds

Northern corn leaf blight, caused by Exserohilum turcicum (Et), is a disease of widespread occurrence in regions where corn, sweetcorn and popcorn are grown. This disease has great potential to cause damage and has been studied for years, but the association of its causal agent with seeds remains unconfirmed. Thus, the availability of a sensitive method to detect and quantify the inoculum in seeds, even at low incidence, is essential. The aim of this study was to develop a method to detect and quantify the presence of the fungus infecting and infesting corn and popcorn seeds. Artificially and naturally infected seeds were employed to develop the medium. The semi-selective medium was composed of carbendazim (active ingredient) (60 mg/L), captan (30 mg/L), streptomycin sulfate (500 mg/L) and neomycin sulfate (600 mg/L) aggregated to the medium lactose casein hydrolysate agar medium. By using this, Et was detected in naturally infected corn seeds, showing 0.124% incidence, in four out of ten analyzed samples. In addition, 1.04 conidia were detected per infested seed. By means of isolation, pathogenicity test, morphological characterization and comparison with descriptions of the species in the literature, the fungus isolated from the seeds was confirmed to be Et. Both infection and infestation were considered low; thus, for studies of Et detection in corn seeds, the use of semi-selective medium and more than 1,200 seeds/sample is suggested.

Primary metabolite mobilization during germination in rosewood (Aniba rosaeodora Ducke) seeds

This study aimed to characterize protein, oil, starch and soluble sugar mobilization as well as the activity of alpha-amylase during rosewood seed germination. Germination test was carried out at 25°C and the following parameters were analyzed: percentage of germination, initial, average, and final germination time. Seed reserve quantification was monitored in quiescent seeds and during different stages of radicle growth. Starch mobilization was studied in function of a-amylase activity. Germination reached 87.5% at the initial, average, and final time of 16, 21 and 30 days, respectively. Oil mobilization showed a negative linear behavior, decreasing 40% between the first and the last stage analyzed, whereas protein levels increased 34.7% during the initial period of germination. Starch content (46.4%) was the highest among those of the metabolites analyzed and starch mobilization occurred inversely to the observed for soluble sugars; alpha-amylase activity increased until the 15th day, a period before radicle emission and corresponding to the highest starch mobilization. The high percentage of rosewood seed germination may be related to the controlled condition used in the germination chamber as well as to high seed reserve mobilization, in special oil and starch.

Histochemical aspects of reserves mobilization of Caesalpinia peltophoroides (Leguminosae) seeds during germination and seedlings early growth

The objective of this study was to investigate reserve mobilization in Caesalpinia peltophoroides seeds during germination and initial seedling growth. The variation in these compounds was analyzed from the pre-germination period (0 to 5 days after sowing - DAS) to the total cotyledon senescence and abscission at 35 DAS. For this histochemical tests were made on cotyledons fixed in FAA50 or included in glycol-metacrylate. To follow the mobilization of the main reserve compounds, sudan III was used to detect total lipids, xylidine Ponceau to detect total proteins, lugol to detect starch and polarized light to visualize the crystals. The lipids, present in a great quantity in the cotyledon, gradually decreased in the period studied. A greater quantity of starch was observed on the 10th DAS than in the previous periods and it was totally consumed by 30 DAS. The distribution pattern and the morphology of the protein material were very modified by 10 DAS, a period during which it was intensely consumed, remaining only parietally fragments distributed, that practically disappeared at 25 DAS. The calcium oxalate druses were not consumed during the period studied, there was only crystal agglutination.

Evaluation of the physiological quality of anadenenthera peregrina (L.) Speg. seeds during storage

The aim of this experiment was to evaluate the effect of storage temperature on the viability and vigour of A. peregrina seeds. Seeds of A. peregrina harvested in Viçosa-MG, in September 2005 were used. The seeds were stored in the Laboratory of Forest Seeds Analysis (LASF) of the Universidade Federal de Viçosa (UFV), in a cold chamber, under approximately 5ºC over150 days. The seeds were stored under 20ºC for 150 days more. Seed moisture was determined after oven-drying at 105 ± 3ºC for 24 hours. The seeds were analyzed monthly using germination, tetrazolium and electrical conductivity tests and Osmopriming during 300 days, with four repetitions of 25 seeds. A. peregrina seeds kept a constant percentage of germination and viable seeds under the tetrazolium test for five months of storage, under either 5ºC and 20ºC. The electrical conductivity test showed significant differences in relation to the storage period, proving to be more sensitive than the standard germination test. Seed priming in PEG -0,4 MPa solution was not efficient in recovering the viability and vigor of the stored seeds at 5ºC and 20ºC.

Histochemical and ultrastructural study of Caesalpinia peltophoroides Benth. (Leguminosae-Caesalpinoideae) seeds

The objective of this work was to correlate data on light microscopy observations through histochemical analysis and polarized light techniques and investigations in transmission electron microscopy (TEM) to characterize the reserve materials in C. peltophoroides Benth. (Leguminosae-Caesalpinoideae) cotyledons, popularly known as "sibipiruna", a tropical tree species with wide distribution in Brazil. The cotyledon mesophyll, especially in the abaxial face, is rich in unsaturated neutral lipids contained in numerous lipid bodies dispersed in the cytoplasm. Proteins, more concentrated in the adaxial face of the cotyledons, occur in all the mesophyll and are stored in protein bodies containing globoids, with variable number and size, responsible for accumulation of mineral reserves. Calcium oxalate druses have distribution restricted to the cotyledons adaxial face and are associated with protein bodies. Starch, also distributed all over the cotyledon mesophyll, occurs in small amounts in plastids with developed lamellar system. Secretory cavities rich in phenolic compounds occur among procambial strands.

Germination of Caesalpinia leiostachya (benth.) ducke seeds classified by size and shape

Caesalpinia leiostachya (Benth.) Ducke (Caesalpiniaceae) is a Brazilian legumenous tree whose seeds present different sizes and shapes, with dormancy imposed by a waterproof coat. In order to improve that quality of lot, seeds were classified by width in sieves of circular meshes (13, 14, 15, 16 and 17) and manually separated by shape in elongate, round and angular. Masses of 1000 seeds were determined and the thickness of each width and shape seedclass was measured, and coat permeability of the seeds classified by the width was investigated. Seeds were scarified in concentrated sulphuric acid and submitted to the germination test. Both final percentage and speed of germination index were evaluated. The classification of the seeds improved the quality of the lot, and the coat permeability was not affected by seed width. Elongated-flat seeds are of low physiological quality and should be discarded. Masses of 1000 seeds, percentage final and speed of germination increased with the width of the seeds. Both elongated and angular seeds had similar germinative behavior, and round seeds are of greater thickness and of superior physiological quality.

Germination and vigor of Dimorphandra mollis benth. seeds under different temperatures and substrates

The Dimorphandra mollis Benth. - Caesalpiniaceae is a native forest species coming from the Cerrado and Caatinga due to its economical and ecological use, which justifies the studies on seed germination. In this work, germinative performance of D. mollis seeds were studied in different conditions of temperature regime and substrate. The experimental delineation used was completely randomized in factorial 4 x 4 (4 substrates -sand, coconut fiber, vermiculite and paper towel; and 4 temperatures: 25, 30, 35 and 20-30ºC), with four replications of 25 seeds each. The following parameters were evaluated: seed moisture content, final germination, first germination count, germination speed index, length and dry matter weight. The best germination and vigor is obtained at 30 and 35ºC. The substrates paper towel and vermiculite allow satisfactory germinative performance of seeds, being suitable to evaluate the physiological quality of D. mollis seeds.

Physiological quality of Gliricidia sepium (Jacq.) Steud. (Leguminosae - Papilionoideae) seeds subjected to different storage conditions

Studies of seed physiological processes represent the starting point for the sustainable utilization of native or cultivated plant species from the caatinga biome for which germination studies are still scarce. In order to determine adequate methods for the conservation of Gliricidia sepium seeds, the physiological quality of propagules exposed to different conditions and times of storage were examined. Recently collected seeds of G. sepium were analyzed immediately, while the other sample was stored for three, six, nine, or twelve months in plastic or paper bags under refrigeration or at room temperature for further analysis. The germination rates of seeds from each storage regime were determined and these rates compared to their water content. It was observed that germination capacity was directly related to changes in seed humidity. Storing seeds in plastic bags yielded the best conservation, mainly when they were stored at low temperatures. However, even at temperatures near 25°C it was possible to prolong seed viability when their water content was controlled through the use of impermeable packing.

REPEATABILITY OF FRUITS AND SEEDS PRODUCTION AND SELECTION OF BRAZIL NUT GENOTYPES IN NATIVE POPULATIONS IN RORAIMA1

ABSTRACT This study estimates the repeatability coefficients of two production traits in two native populations of Brazil nut trees. It determines the number of years of suitable evaluations for an efficient selection process, determines the permanent phenotypic correlation between production traits and also the selection of promising trees in these populations. Populations, located in the Itã region (ITA) and in the in the Cujubim region (CUJ), are both belonging to the municipality of Caracaraí, state of Roraima - Brazil, and consist of 85 and 51 adult trees, respectively. Each tree was evaluated regarding the number of fruits per plant (NFP) and fresh seed weight per plant (SWP), for eight (ITA) and five consecutive years (CUJ). Statistical analyses were performed according to the mixed model methodology, using Software Selegen-REML/BLUP (RESENDE, 2007). The repeatability coefficients were low for NFP (0.3145 and 0.3269 for ITA and CUJ, respectively) and also for SWP (0.2957 and 0.3436 for ITA and CUJ, respectively). It on average takes nine evaluation years to reach coefficients of determination higher than 80%. Permanent phenotypic correlation values higher than 0.95 were obtained for NFP and SWP in both populations. Although trees with a high number of fruits and seed weight were identified, more evaluation years are needed to perform the selection process more efficiently.