Pathogenicity, molecular characterization, and cercosporin content of Brazilian isolates of Cercospora kikuchii

Cercospora kikuchii, involved with the defoliation of soybean (Glycine max) plants, is normally associated with Septoria glycines in late season. Seventy-two isolates from different regions in Brazil, obtained mainly from purple stained seeds, showed phenotypic variation. Cercosporin content and rate of colony growth was higly variable among isolates. A strong correlation between cercosporin content and virulence was identified. Genetic variation among and within populations was evaluated based on 86 RAPD loci. The RAPD analysis clustered all isolates into seven groups. No relationship was observed between RAPD groups and geographic origin or cercosporin content. The sequence of the internal spacer regions (ITS1-5.8S-ITS2) from 13 isolates chosen according to the previous RAPD and clustering analysis showed high similarity (97%-100%) to the GenBank sequences of C. kikuchii (AY266160, AY266161, AY152577 and AF291708). It is clear from this work that Brazilian isolates of C. kikuchii from different geographic regions, are variable in relation to virulence, RAPD profiles and cercosporin content. Cercosporin content could be a good parameter for choosing an adequate isolate for screening resistant or tolerant cultivars. Considering that this pathogen is easily seed-borne, findings are expected to show the same haplotypes in different regions. Migration could be favoured by infected seeds as demonstrated by RAPD analysis.

Allelopathic potential and systematic evaluation of secondary compounds in extracts from roots of Canavalia ensiformis by capillary electrophoresis

Organic extracts were obtained from roots of Canavalia ensiformis and evaluated for allelopathic potential on the germination of the weed seeds: Mimosa pudica, Cassia tora and Cassia occidentalis showing a strong allelopathic potential. After that, a systematic study of these crude extracts was made using specific protocols developed in capillary electrophoresis (CE) in order to determine some classes of secondary metabolites. Capillary electrophoresis protocols were highly specific, which makes it possible to identify 5 classes of compounds using the same crude extract samples and analyze them fartly. Some of the compounds identified show activity in the inhibition of seeds germination.

Cotton resistance to ramulose and variability of Colletotrichum gossypii f. sp. cephalosporioides

Four cultivars and 21 lines of cotton were evaluated for resistance to ramulose (Colletotrichum gossypii f. sp. cephalosporioides) in a field where the disease is endemic. The seeds of each genotype were planted in 5 x 5 m plots with three replications. The lines CNPA 94-101 and 'CNPA Precoce 2'were used as standard susceptible and resistant references, respectively. The disease incidence (DI) was calculated from the proportion of diseased plants in the plot. The disease index (DIn) was calculated from the disease severity using a 1 to 9 scale, and was evaluated at weekly intervals starting 107 days after emergence. The data collected was used to calculate the area under disease progress curve (AUDPC). In general, the DIn increased linearly with time and varied from 20.0 to 57.1 and AUDPC from 567 to 1627 among the genotypes which could be clustered in to two distinct groups. The susceptible group contained two cultivars and nine lines and the resistant group contained one cultivar and 12 lines. The relationship between disease index and evaluation times was linear for the 25 genotypes tested. The line CNPA 94-101, used as susceptible standard, was the most susceptible with an average DI = 83.4, DIn = 57.1 and AUDPC = 1627.7. The line CNPA 96-08 with DI = 37.8, DIn = 20.0 and AUDPC = 567.7 was the most resistant one. Among the commercial cultivars 'IAC 22' was the most susceptible and 'CNPA Precoce 2', used as resistant standard was the most resistant. The variability in virulence of the pathogen was studied by spray inoculating nine genotypes with conidial suspensions (10(5)/mL) of either of the 10 isolates. The disease severity was evaluated 30 days later using a scale of 1 to 5. The virulence of the isolate was expressed by DIn. All the isolates were highly virulent but their virulence avaried for several genotypes and could be clustered in two distinct groups of less and more virulent isolates. The isolate MTRM 14 from Mato Grosso was the least virulent while Minas Gerais was the most virulent, with DIn of 6.36 and 46.47, respectively. In this experiment the line HR 102 and the cultivar 'Antares' were the most resistant ones with DIns of 18.32 and 19.14, respectively.

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.

Susceptibility of leguminous green manure species to Rhizoctonia solani and Sclerotium rolfsii

We studied the susceptibility of species used as green manure in common bean fields to root rot (Rhizoctonia solani) and southern blight (Sclerotium rolfsii). Seeds of Crotalaria breviflora, Canavalia ensiformis, Cajanus cajan, Dolichos lablab, Stizolobium cinereum, S. aterrimum, and the bean cvs. "Pérola", "Valente" and "Carnaval" were sown in soil infested by either R. solani AG-4 or S. rolfsii in greenhouse. The emergence of D. lablab seedlings in soil infested by R. solani dropped to 62%. C. breviflora, C. ensiformis and cv. "Valente" presented the lowest root rot severity. The pathogen S. rolfsii drastically reduced seedling emergence in all species; no C. cajan and S. cinereum seedling emerged. All plant species presented high southern blight severity. We conclude that leguminous crops are not suitable as green manure for areas of bean cultivation with high R. solani and S. rolfsii populations.

Use of botanical extracts, cassava wastewater and nematicide for the control of root-knot nematode on carrot

Aqueous extracts of several plant species have shown promising in controlling root-knot nematode, Meloidogyne incognita (Kofoid & White), one of the most limiting agents for carrot cultivation. The current study evaluated the effect of aqueous extracts from seven botanical species applied to 40, 50, 60, 70 and 80 days after sowing 'Nantes' carrots in soil infested with root-knot nematode. Three other treatments included cassava wastewater, distilled water (control), which were applied in the same periods of the extracts application, in addition to carbofuran 50G (80Kg/ha), which was applied once at 60 days after carrot sowing. Evaluations were performed at 90 days after inoculation to determine shoot and root fresh weight, as well as the diameter and the length of principal roots and the number of galls on primary and secondary roots. Plants treated with cassava wastewater, extracts of Ricinus communis L. seeds, Crotalaria juncea L. seeds, R. communis leaves + branches + fruits, Chenopodium ambrosioides L. leaves + branches + inflorescences and Azadirachta indica A. Juss. seeds showed the highest rates of total weight (root + shoot) and shoot weight. The extract of R. communis leaves + branches + fruits provides the highest total root weight and principal root diameter. Cassava wastewater and extracts of R. communis seeds provided the highest principal root weight. The extract of R. communis seeds and cassava wastewater can be considered promising for the alternative control of M. incognita.

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.

Seed germination and removal of Michelia champaca L. (Magnoliaceae) in eucalypt stands: the influence of the aril

We tested the influence of the aril on seed germination in controlled conditions and on the removal of M. champaca seeds in natural environment. Germination assays were kept at 25 ± 2 °C under continuous white light. Removal experiments were carried out in three "old" (39 to 62-years old) and three "new" (15-years old) eucalypt stands in Horto Florestal Navarro de Andrade, Rio Claro, SP. The results show that the aril inhibits the germination and the seeds exhibit a positively photoblastic reaction. We found higher seed removal in old eucalypt stands than the new ones, probably due to the higher density of rodents in the old stands. In the new stands, we found higher seed removal of arillated seeds by ants. Ants are important to remove the aril of seeds dropped by birds, not only enhancing seed germination but also preventing seed predation by rodents.

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.

Reproductive ecology of the exotic tree Muntingia calabura L. (Muntingiaceae) in southeastern Brazil

The exotic tree Muntingia calabura L. (Muntingiaceae), a species native to Central America, is used as fish feed and fiber and cellulose production in Brazil. This study was carried out in urban areas and verified the reproductive biology of this plant species. Flower and fruit morphology, compatibility system, reproductive phenology, pollination and frugivore animals, and germination of disseminated seeds were recorded by standard field and laboratory procedures. This tree is self-compatible and autonomously self-pollinated, with its flowers being mainly visited by bees and its fruits consumed by birds and bats. Germination of its dispersed seeds is fast and occurs at a high rate. The results of this work suggest that M. calabura is very adaptable to cultivated areas,thus being an excellent choice for urban reforestation. However, its reproductive characteristics place this plant as an invasive species with significant potential in southeastern Brazil.

Caesalpinioideae (leguminosae) lenhosas na Estação Ambiental de Volta Grande, Minas Gerais, Brasil

Este trabalho apresenta o levantamento florístico das Caesalpinioideae lenhosas nas formações de Cerrado e de Floresta Semidecidual, da Estação Ambiental de Volta Grande. A área de estudo, localizada no Triângulo Mineiro, faz parte do complexo da Usina Hidrelétrica Estadual de Volta Grande, reúne 391 ha e retrata 30 anos de regeneração natural. Foram registrados 14 táxons da subfamília, reunidos em 11 gêneros e quatro tribos. Caesalpinieae foi a tribo mais representada (Dimorphandra Schott, Diptychandra Tul, Peltophorum (Vogel) Benth., Pterogyne Tul. e Tachigali Aubl.), seguida por Cassieae (Apuleia Mart., Chamaecrista Moench e Senna Mill.), Detarieae (Copaifera L. e Hymenaea L.) e Cercideae (Bauhinia L.). O gênero mais representativo foi Senna (4 spp.), enquanto os demais foram representados por uma espécie cada. Apresentam-se chave para identificação, descrições e ilustrações, além de comentários sobre a distribuição geográfica dos táxons encontrados.

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.

Seed germination of a rare neotropical canopy tree dormancy and the effects of abiotic factors

The purpose of this study was to examine if germination is a critical phase on Enterolobium glaziovii regeneration. Hence, the germinative response of E. glaziovii seeds was investigated in relation to some of the main environmental factors (temperature, light and water stress) to which its seeds are subjected in the forest, as well as its dormancy and the longevity of its burial seeds. According to our results, its seeds may be regarded as photoblastic neutral. They do not need alternating temperatures to germinate and can germinate under a broad range of water stress. However, only about 10% of E. glaziovii seeds remain viable after one year. In other words, the annual fruiting, instead seed longevity, seems to maintain the long-term seed availability of this species. Consequently, the seed longevity could be a critical phase of E. glaziovii germination.

Physiological and biochemical characterization of the assai palm (Euterpe oleracea Mart.) during seed germination and seedling growth under aerobic and anaerobic conditions

Physiological and biochemical aspects of assai palm during seed germination and early seedling growth were investigated. Seeds collected from plants growing in flooded and upland forests were used to determine the influence of normoxic (aerobic) and anoxic (anaerobic) conditions in germination and the initial and average time of development in the roots and shoots. After 75 days, seedlings germinated under normoxia were transferred to trays and submitted to flooding. Seed reserves (lipids, proteins, soluble sugars and starch) were monitored for quiescent and germinated seeds maintained under normoxic and anoxic conditions, as well as after 5, 10 and 20 days of seedling growth. Alcohol dehydrogenase (ADH) activity was quantified in roots and leaves of seedlings without or with flooding (partial and total). Seeds were not able to germinate under anoxia. Different strategies of storage mobilization of lipids, proteins, soluble sugars and starch were observed in seeds of each environment. ADH activity was induced by anoxia, with the highest level observed in the leaves. This study showed that, under normoxic conditions, the best developmental performance of assai palm seeds, from flooded or upland forest areas, during germination was associated with primary metabolites mobilization and seedling flooding tolerance with increased ADH activity. We conclude that the assai palm is well adapted to the anoxic conditions provoked by flooding.

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.