Effect of organic amendments on sclerotial germination, mycelial growth, and Sclerotium rolfsii-induced diseases

The addition of organic residues to soil is an option to control some soil-borne diseases. Benzaldehyde and powders of kudzu (Pueraria lobata), velvetbean (Mucuna deeringiana), and pine-bark (Pinus elliottii and P. taeda) added to soil could reduce certain soil-borne diseases. This study evaluated the effects of benzaldehyde and the dried powders of kudzu, velvetbean, and pine-bark as soil amendments on germination and formation of sclerotia, on mycelial growth of Sclerotium rolfsii, on plant survival, and disease incidence. The data showed that high amounts of benzaldehyde (0.4 ml kg-1 of soil) and velvetbean (100 g kg-1) inhibited S. rolfsii mycelial growth and sclerotium germination. However, low amounts of benzaldehyde (0.1 ml kg-1), kudzu (25 g kg-1), and pine-bark (25 g kg-1) stimulated mycelial growth and sclerotium germination. Kudzu (25-100 g kg-1) and velvetbean (25-100 g kg-1) inhibited the formation of sclerotia. Nevertheless, benzaldehyde at 0.2 and 0.4 ml kg-1 stimulated the formation of sclerotia. Kudzu (50 and 100 g kg-1) and pine-bark (50 g kg-1) favored the colonization of sclerotia by Trichoderma sp. The numbers of soybean (Glycine max) plants were higher and diseased plants were lower than the non-amend soil in the following treatments: kudzu (50 and 100 g kg-1), velvetbean (50 and 100 g kg-1), and pine-bark (50 g kg-1). Disease severity on tomato (Lycopersicon esculentum) plants was low in soil treated with kudzu or velvetbean (30 and 35 g kg-1) and pine-bark (35 g kg-1). Dried powders of kudzu, velvetbean, or pine-bark added to soil can reduce disease by reducing pathogen inoculum.

Screening of sweet sorghum accessions for inhibition of secondary sporulation and saccharide measurements in honeydew of Claviceps africana

Certain sweet sorghums (Sorghum bicolor) inhibit the secondary sporulation of Claviceps africana, which occurs on exuded ergot honeydew when the parasite is supplied with excess sucrose, which is then transformed to unique free oligosaccharides fructosyl - mannitol and difructosyl - mannitol with spore germination inhibiting properties. Five accessions (BRA-035726-SUGAR DRIP, BRA-035696-THEIS, BRA-036013-MN-4578, BRA-035947-MN-4418 and CMSXS-633) of sweet sorghum were selected among 50 evaluated. These five accessions failed to support secondary sporulation on the "honeydew" exuded from infected florets. There was a higher concentration (%w/v) of the free oligosaccharides on the honeydew of these accessions when compared to a hybrid male-sterile grain sorghum. Therefore, a possible strategy would be seek to incorporate a sweet character into "A" lines for hybrid seed production in order to restrict secondary disease spread.

Detection and transmission of Drechslera avenae from oat seed

Detection, symptoms and symptomless transmission of Drechslera avenae (teleomorph Pyrenophora avenae) from seed were investigated. The present study reported that the D. avenae is frequently found in oat (Avena sativa) seed in Argentina. The prevalence of the seed lot infected was 54%. The incidence of seeds infected by D. avenae in the various seed lots from different regions ranged from 0 to 52% (overall mean of 7%). Besides conidia on conidiophore and immature pseudothecia, D. avenae produced small, spherical to pear-shape picnidia containing tiny conidia, on the seed surface. The pathogen was efficiently transmitted at a rate of 55% from seed to coleoptile tips in symptomless seedlings and at 12.5% to plumules. The importance of the infected seed and its epidemiological role are discussed.

A Semi-Selective agar medium to detect the presence of Xanthomonas axonopodis pv. malvacearum in naturally infected cotton seed

A semi-selective agar medium was developed for detection of Xanthomonas axonopodis pv. malvacearum (Xam) in cotton (Gossypium hirsutum) seed. The basic medium was peptone-sucrose-agar (PSA). Criteria for the semi-selective medium were the typical colony characters of Xam and its pathogenicity on cotton. Several systemic fungicides and antibiotics in different concentrations were tested alone or in combination with others. The final composition of the semi-selective agar medium was established after several attempts in order to inhibit most of the fungal and bacterial saprophytes and favour the development of Xam. It contained PSA + cyclohexamide, cephalexin, pencycuron, triadimenol and tolylfluanid. The bacteria were recovered from naturally infected seeds by the direct plating of 2,000 surface disinfected seeds on the semi-selective medium. The recovery of the pathogen from naturally infected leaf tissues and in dilution plating, on semi-selective medium and on nutrient agar, were comparable. Among the three detection methods tested, the semi-selective medium was found to be the most reliable and quantifiable. Degree of severity of angular leaf spot in the field was not always correlated with the level of infection in the seed. This is the first report of a semi-selective agar medium to detect the presence of Xam in naturally infected cotton seed.

The structures of the honeydew oligosaccharides synthesized by Claviceps africana

The structures of the principal oligosaccharides in the honeydew exudate of the sorghum ergot pathogen Claviceps africana, which has become epidemic in the Americas, have been characterized through linkage analysis using FAB-MS and GC-MS techniques, as 1,6-di-b-D-fructofuranosyl-D-mannitol and 1,5-di-b-D-fructofuranosyl-D-arabitol trisaccharides, 1-b-D-fructofuranosyl-D-mannitol and 5-b-D-fructofuranosyl-D-arabitol disaccharides and other minor disaccharides and trisaccharides. Their structural diversity is explained according to perceived biosynthetic interrelationships in pathways that appear to be unique amongst ergot fungi, particularly concerning intra-molecular reduction of fructose. The oligosaccharide, 1,6-di-b-D-fructofuranosyl-D-mannitol, which inhibits C. africana macrospore germination at a concentration in water of 1 g/mL or more, forms together with other slightly less bioactive oligosaccharides, the basis of a novel strategy to limit ergot disease losses in hybrid sorghum seed production.

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.

Semi-selective medium for Fusarium graminearum detection in seed samples

Fungi of the genus Fusarium cause a variety of difficult to control diseases in different crops, including winter cereals and maize. Among the species of this genus Fusarium graminearum deserves attention. The aim of this work was to develop a semi-selective medium to study this fungus. In several experiments, substrates for fungal growth were tested, including fungicides and antibiotics such as iprodiona, nystatin and triadimenol, and the antibacterial agents streptomycin and neomycin sulfate. Five seed samples of wheat, barley, oat, black beans and soybeans for F. graminearum detection by using the media Nash and Snyder agar (NSA), Segalin & Reis agar (SRA) and one-quarter dextrose agar (1/4PDA; potato 50g; dextrose 5g and agar 20g), either unsupplemented or supplemented with various concentrations of the antimicrobial agents cited above. The selected components and concentrations (g.L-1) of the proposed medium, Segalin & Reis agar (SRA-FG), were: iprodiona 0.05; nystatin 0,025; triadimenol 0.015; neomycin sulfate 0.05; and streptomycin sulfate, 0.3 added of ¼ potato sucrose agar. In the isolation from seeds of cited plant species, the sensitivity of this medium was similar to that of NSA but with de advantage of maintaining the colony morphological aspects similar to those observed in potato-dextrose-agar medium.

Sensitivity reduction in Blumeria graminis f. sp. hordei to triadimenol fungicide applied as barley seed treatment

Experiments were carried out in a growth chamber with controlled temperature and photoperiod to test two populations of Blumeria graminis f. sp. hordei from Guarapuava, Paraná State, and Passo Fundo, Rio Grande do Sul State, Brazil. Treatments consisted in application of the fungicide triadimenol (Baytan 150 SC®) at three rates of its commercial formulation: 150, 250, 350 mL/100 Kg barley seeds. The experiments were conducted separately in a growth chamber for each population, adopting the same temperature and photoperiod. For inoculation, pots containing barley seedlings colonized by the fungus were placed among the plots. After emergence of the first symptoms, the disease severity was assessed at two-day intervals. The experiments were repeated twice for each fungus population. Data were expressed as area under the disease progress curve and as powdery mildew control by comparing the severity after the fungicide treatments to that of control. Data were subjected to analysis of variance and regression analysis; the area under the disease progress curve was also calculated. Comparing the data obtained in the present study with those reported in the literature and the control, the maximum value of 26.1% is considered insufficient to prevent the damages caused by the disease. The control response to the fungicide rate was significant. We can conclude that there was a reduction in the sensitivity of both B. graminis f.sp. hordei populations to the fungicide triadimenol, which explains the control failure observed in barley farms.

In vitro effect of substrate, temperature and photoperiod on Phakopsora pachyrhizi urediniospore germination and germ tube growth

In vitro experiments were conducted to assess the effects of substrate, temperature and time of exposure to temperature and photoperiod on P. pachyrhizi uredospore germination and germ tube growth. The following substrates were tested: water-agar and soybean leaf extract-agar at different leaf concentrations (0.5, 1.0, 2.0 and 4.0 g of leaves and 15g agar/L water), temperatures (10, 15, 20, 25, 30, and 35oC) and times of exposure (1, 2, 3, 4, 5, 6, 7, and 8 hours) to temperature and 12 different photoperiods. The highest germination and germ tube length was found for the soybean leaf extract agar. Maximum P. pachyrhizi uredospore germination was obtained at 21.8 and 22.3°C, and maximum germ tube growth at 21.4 and 22.1°C. The maximum uredospore germination was found at 6.4 hours exposure, while the maximum germ tube length was obtained at 7.7 h exposure. Regarding photoperiod, the maximum spore germination and the maximum uredospore germ tube length were found in the dark. Neither spore germination nor uredospore germ tube growth was completely inhibited by the exposure to continuous light.

Secondary seed dispersal of Ricinus communis Linnaeus (Euphorbiaceae) by ants in secondary growth vegetation in Minas Gerais

In this study, I tested the efficacy of ants as secondary seed dispersers of Ricinus communis in southeastern Brazil. In a natural population of 143 individuals, I determined the ballistic dispersal distance for 62 seeds and 100 additional seeds were experimentally offered to ants in groups of ten seeds along a transect of 50 m. Fifty-three seeds were removed by ants, mainly by the leafcutter Atta sexdens (90.4%). The dispersal distance by ants was high, compared to the global average (4.38 m ± 0.74 m vs. 0.96 m), but was lower than the ballistic distance (7.27 m ± 0.13 m). Ants increased the total dispersal distance (8.66 m ± 0.60 m), but the main benefit for the plant was the directed dispersal, with seed deposition on the enriched soil of ant nests.

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.

Micropropagation of Maclura tinctoria L.: an endangered woody species

Some native species produce seeds with low germination percentage and in most cases with dormancy, which makes the appearance of new individuals by sexual propagation difficult. The Maclura tinctoria has been considered an endangered species due to the indiscriminate use of its wood and low rate of seed germination. In this context, the objective of the present study was to establish an in vitropropagation methodology for this species. Combinations of NAA + BAP, different concentrations of GA3 and combinations IBA + activated charcoal were evaluated for shoot induction, shoot growth and root formation, respectively. The results indicated that the maximum shoot formation was obtained when 5.37 µM NAA + 4.45 µM BAP was used. The use of 5.48 µM GA3 promoted shoot growth. Root formation was observed on explants inoculated in WPM with a pH adjusted to 7.0 and supplemented with 23.62 µM IBA + 4.7 g L-1 activated charcoal. The use of a 70% light screen for 7 days followed by the use of 50 and 30% light screens also for 7 days each provided 97% plantlet survival.

Comparisons between two economically valuable forest species (Eucalyptus grandis and Pinus taeda) in relation to seed behaviour under controlled deterioration

The objectives of this work were to analyze seed behaviour under controlled deterioration and estimate viability equations for forest species Eucalyptus grandis and Pinus taeda. Desired moisture content levels were achieved from initial values after either rehydration over water or drying over silica gel, both at 25 ºC. Seed sub samples with 8 moisture contents each for E. grandis (1.2 to 18.1%, initial value of 11.3%) and P. taeda (1.5 to 19.5%, initial value of 12.9%) were sealed in laminate aluminium-foil packets and stored in incubators maintained at 40, 50 and 65 ºC. The seeds from these species exhibited true orthodox and sub-orthodox storage behaviour, respectively, however E. grandis showed higher seed storability, probably due to a different seed chemical composition. Lowest moisture content limits estimated for application of the viability equations at 65 ºC were 4.9 and 4.1 mc for E. grandis and P. taeda, on equilibrium with ±20% RH. The viability equation estimated quantified the response of seed longevity to storage environment well with K E = 9.661 and 8.838; C W = 6.467 and 5.981; C H = 0.03498 and 0.10340; C Q = 0.0002330 and 0.0005476, for E. grandis and P. taeda, respectively.

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.