ABSTRACT Harvest delay can alter the physiological and sanitary quality of sorghum seeds due to the longer exposure time of seeds to adverse environmental conditions in the field. In this study, we investigated the impact of delayed Sorghum sp. harvest on the physiological and sanitary quality of seeds in four hybrids grown in the tropical soil of the Brazilian Cerrado region. Plants from four Sorghum sp. hybrids (IPA SUDAN 4202, BRS 658, BRS 810, and BRS 373) were grown and submitted to four seed harvest periods (0, 7, 14, and 21 days after stage 9 - seed physiological maturity). The treatments were arranged in a completely randomized block design in a 4 × 4 factorial, with four replicates. At each harvest period, the thousand-seed weight, water content, first count of the germination test, germination, seedling emergence, emergence speed index, seedling dry matter, accelerated aging, electrical conductivity, and sanitary quality of seeds were determined. Seeds of higher physiological and sanitary quality of the IPA SUDAN 4202 and BRS 658 sorghum hybrids can be obtained when the harvest is carried out up to 7 days after the physiological maturity stage. However, the delay in seed harvesting at 14 and 21 days after physiological maturity results in a higher incidence of Colletotrichum sp., Curvularia sp., Rhizoctonia sp., and Alternaria sp. on the seeds of these hybrids. Delaying the harvest of sorghum seeds from 7 days after the physiological maturity stage compromises seed vigor and viability and increases the incidence of pathogens.

Resumo Nas regiões Centro-Oeste e Sul do Brasil, a cultura da erva-mate (Ilex paraguariensis) constitui-se uma importante fonte de extrativismo florestal, além de diversificar a renda, principalmente na agricultura familiar. A erva-mate, como outras espécies florestais, é infectada por diversos patógenos, como o gênero Fusarium, causador da podridão-de-raízes. Os patógenos possuem ampla variabilidade genética, mesmo dentro de um único gênero e dentro de uma única espécie, resultado de adaptações ao meio em que vivem. Este estudo teve como objetivo avaliar o sequenciamento das regiões do genoma ITS e β-tubulina na identificação em nível de espécie dos isolados de Fusarium spp. patogênicos à erva-mate, e identificar a formação de grupos de compatibilidade vegetativa (VCG) entre os isolados de Fusarium spp. patogênicos à erva-mate. O sequenciamento das regiões ITS e β-tubulina consolidou a separação entre os sete isolados patogênicos em F. solani e F. oxysporum. Para esses isolados, a técnica para obtenção de mutantes nit foi adequada para a classificação fenotípica, fundamentais nos testes de VCG para espécies de Fusarium, sendo obtidos cinco VCG.

Abstract In the central-western and southern regions of Brazil, the cultivation of yerba mate (Ilex paraguariensis) is considered an important source of forest extraction, besides providing an alternative income, especially for families. The yerba mate, like other forest species, is infected by several pathogens, such as fungi of the genus Fusarium, causing root rot. Pathogens have wide genetic variability, even within a single genus and species, resulting from adaptations to their environments. This study aimed at characterizing molecularly isolated Fusarium spp. through the sequencing of the ITS and β-tubulin genome regions, identifying the species level as well as the formation of vegetative compatibility groups (VCG) among these isolates of the root rot pathogen in yerba mate. The sequencing of the ITS and β-tubulin regions, defined the separation between the seven pathogenic isolates of F. solani and F. oxysporum. For the same isolates, the technique to obtain nit mutants was suitable for the phenotypic classification, obtaining five VCGs for species of Fusarium, root rot in yerba mate.

RESUMO A erva-mate (Ilex paraguariensis) tem grande importância socioeconômica na agricultura familiar do Sul do Brasil. A principal doença da erva-mate é a podridão-de-raízes causada por Fusarium spp. Pouco se sabe a respeito da fisiologia deste patógeno, principalmente quanto à agressividade associada à produção de enzimas extracelulares. Neste trabalho, avaliou-se a agressividade de isolados de F. oxysporum e F. solani patogênicos à erva-mate e determinou-se as atividades das enzimas extracelulares catalase, lacase, celulase, caseinase, amilase, protease, lipase e pectinases produzidas por Fusarium spp. em meio de cultura. Foram utilizados seis isolados de F. solani e um isolado de F. oxysporum patogênicos à erva-mate. O isolado de F. oxysporum mostrou-se menos agressivo em relação aos demais isolados de F. solani. Todos os isolados de Fusarium spp. produziram, de maneira semiquantitativa, as enzimas extracelulares catalase, lacase, celulase, caseinase, amilase, protease, lipase e pectinases (poligalacturonase e pectato liase). Entretanto, a quantidade produzida de cada enzima foi significativamente diferente entre os isolados. Com exceção das enzimas lacase e poligalacturonase o isolado M7C1 de F. solani exibiu as maiores médias do índice enzimático e foi também o responsável pelo maior percentual de morte de mudas de erva-mate.

ABSTRACT The yerba-mate (Ilex paraguariensis) has great socioeconomic importance on family farming in Southerm Brazil. One of the main yerba-mate disease is root rotting, caused by Fusarium spp. Little is known about the pathogen physiology, especially regarding the aggressiveness associated with the production of extracellular enzymes. On this work, the aggressiveness of isolates of F. oxysporum and F. solani pathogenic to yerba-mate was evaluated and it was determined the activities of extracellular enzymes catalase, laccase, cellulase, caseinase, amylase, protease, lipase and pectinases produced by Fusarium spp. in culture medium. Six isolates of F. solani and one isolate of F. oxysporum pathogenic to yerba-mate were used. The F. oxysporum isolate proved to be less aggressive in relation to the other F. solani isolates. All isolates of Fusarium spp. produced, on a semiquantitative manner, the extracellular enzymes catalase, laccase, cellulase, caseinase, amylase, protease, lipase and pectinases (polygalacturonase and pectate lyase). However, the quantity produced for each enzyme was significantly different among the isolates. With the exception of the laccase and polygalacturonase enzymes, the M7C1 isolate showed the highest enzymatic index and was also responsible for the highest percentage of yerba-mate seedlings death.

ABSTRACT Pecan is susceptible to the occurrence of diseases capable of harming its development, and leaf spot caused by Cladosporium spp is among them. Therefore, the objective of this work was to analyze the potential antagonist and in vitro effect of volatile metabolites of Trichodermaspp. On Cladosporium spp. isolated from pecan leaf spots. Three Cladosporium spp. isolates and two commercial products based on Trichodermaspp. were used, as well as an isolate of the same genus provided by the 'ElocyMinussi' Laboratory of Phytopathology at UFSM. Cladosporium spp. mycelial growth was monitored in order to evaluate the morphological characteristics of the pathogen. The use of Trichoderma spp. as biocontrol was assessed through dual culture tests and production of volatile metabolites. Trichoderma spp. showed positive results in inhibiting the growth of Cladosporium spp., thereby presenting potential to be used as a biocontrol agent.

RESUMO O objetivo deste trabalho foi avaliar o comportamento de isolados de Ceratocystis fimbriata obtidos de kiwi (Actinidia deliciosa) em mudas de acácia-negra (Acacia mearnsii De Wild.), o potencial de biocontrole in vitro de Bacillus subtilis sobre C. fimbriata e o comportamento in vivo de B. subtilis contra C. fimbriata em mudas de acácia-negra. Isolados de C. fimbriata obtidos e identificados em plantios de kiwi foram utilizados em testes de patogenicidade em mudas de A. mearnsii. O potencial antagônico in vitro de B. subtilis sobre o patógeno foi analisado em testes de confrontação direta. Para avaliar o comportamento in vivo de B. subtilis, as mudas de acácia-negra foram inoculadas com Rizolyptus® sete dias antes e após a inoculação do patógeno. Nas mudas de acácia-negra os isolados de C. fimbriata exibiram percentuais de severidade que variam de 44,15% a 100%. Nos testes de confrontação direta as cepas mostraram-se eficientes no biocontrole in vitro sobre os isolados de C. fimbriata. In vivo, o produto Rizolyptus® não foi eficiente no controle de C. fimbriata.

ABSTRACT The aim of this study was to evaluate the effect of Ceratocystis fimbriata isolates, obtained from Actinidia deliciosa (kiwifruit), on black wattle (Acacia mearnsii De Wild.) seedlings, the potential biological control exerted in vitro by Bacillus subtilis on C. fimbriata, and the in vivo effect of B. subtilis against C. fimbriata on black wattle seedlings. Isolates of C. fimbriata obtained and identified from kiwi plantations were used in pathogenicity tests on A. mearnsii seedlings. In vitro, the antagonistic potential of B. subtilis on the pathogen was analyzed by direct confrontation tests. To evaluate the effect of B. subtilis in vivo, black wattle seedlings were inoculated with Rhizolyptus® seven days before and after pathogen inoculation. On black wattle seedlings, C. fimbriata isolates caused 44,15% to 100% severity. Direct confrontation tests revealed that the strains were effective in the biological control of C. fimbriata isolates in vitro. In vivo, Rizolyptus® was not efficient in controlling C. fimbriata.

ABSTRACT The cultivation of yerba-mate (Ilex paraguariensis) has been intensified in the State of Rio Grande do Sul, as result of the valuation of the price paid to the producer. However, studies related to Fusarium pathosystem vs. yerba-mate are still insufficient, so that many producers have faced significant losses in herbal production due to the occurrence of the roots rotting. Considering the above, the objectives of this study were: a) morphologically characterize the Fusarium spp. isolates collected in sick plants of yerba-mate; b) select efficient morphological characters on separating the Fusarium spp. isolates in similarity groups; c) identify the Fusarium spp. isolates to species level through the sequencing of the genomic region of elongation factor 1-α (TEF-1α); and d) testing the pathogenicity of the Fusarium spp. isolates selected based on the morphological characterization. For such, collections for the pathogen isolates were made in five cities and the collection points were georeferenced. Thirty-nine isolates were morphologically characterized by variables as mycelial growth, sporulation, macroconidia length and width, microconid format, colonies pigmentation and formation of chlamydospore, and tested for pathogenicity by substrate inoculation. The variables used in the morphological characterization were efficient in discriminating the isolates in seven groups, especially length of macronidia associated to sporulation. Seven pathogenic isolates of Fusarium spp. were identified, also through molecular sequencing of the TEF-1α gene region. The sequencing of the TEF-1α region consolidated the identification of pathogenic isolates as F. solani and F. oxysporum.

RESUMO Cordia americana (L.) Gottschling & J. S. Mill. (guajuvira) é uma espécie florestal amplamente distribuída no território brasileiro e que apresenta importância econômica e ambiental devido à qualidade de sua madeira e utilização em reflorestamentos. Com o objetivo de avaliar a patogenicidade de Fusarium sp. associado às sementes, foram caracterizados, morfológica e molecularmente, quatro isolados do fungo que foram inoculados nas sementes de guajuvira. A caracterização morfológica foi realizada com o auxílio de uma chave de identificação para o gênero Fusarium e para a identificação molecular foram sequenciadas as regiões genômicas Internal Transcribed Spacer (ITS) e Fator de elongação 1 - alpha (TEF1-α). Os resultados do sequenciamento de DNA concordaram com os da caracterização morfológica e permitiram identificar as espécies Fusarium acuminatum e Fusarium verticillioides. No teste de patogenicidade, verificou-se que ambas as espécies do fungo foram patogênicas a Cordia americana, causando tombamento em pré e pós-emergência, além de apodrecimento de raízes e necrose no hipocótilo.

ABSTRACT Cordia americana (L.) Gottschling & J. S. Mill. (guajuvira) is a forest species widely distributed in Brazil and represents environmental and economic importance due to the quality of its timber and use in reforestation. In order to evaluate the pathogenicity of Fusarium sp. associated with seeds, characterizing the isolates morphologically and molecularly in order to identify them on the species level, was obtained four fungal isolates that were inoculated in seeds to demonstrate their pathogenicity. The morphological characterization was performed using an identification key for the genus Fusarium and, for the molecular identification, genomic regions Internal Transcribed Spacer (ITS) were sequenced with its elongation factor 1 alpha (TEF1-α). The results of the molecular identification agreed with the morphological characterization and allowed to identify the species Fusarium acuminatum and Fusarium verticillioides, both being pathogenic to Cordia americana, causing damping-off in pre and post emergence, and rotting of roots and necrosis of hypocotyl.

RESUMO Sementes sadias e rápido desenvolvimento de plântulas de espécies florestais nativas são fundamentais para o seu estabelecimento. Este trabalho avaliou o efeito de Trichoderma spp. sobre sementes e o desenvolvimento de plântulas de Parapiptadenia rigida (angico), Cedrela fissilis (cedro) e Pelthophorum dubium (canafistula). O fungo foi aplicado em diferentes momentos da produção de mudas. Sua ação foi diferenciada entre as espécies, bem como, entre as variáveis analisadas. O cedro foi menos responsivo à aplicação de Trichoderma spp. em qualquer momento da produção de mudas. A associação de Trichoderma spp. com angico, via tratamento de sementes, favoreceu a incidência de alguns patógenos, entretanto a aplicação mais tardia do fungo favoreceu o desenvolvimento da parte aérea. Sobretudo, em canafístula, o tratamento de sementes proporcionou controle de patógenos associados à semente e favoreceu a emergência e a produção de folhas nas mudas, apresentando potencial para ser usado tratamento de sementes desta espécie.

ABSTRACT Seeds healthy and rapid development of seedlings of native species may be critical to the success of your establishment. This study evaluated the effect of Trichoderma spp. on seed and seedling development of Parapiptadenia rigida (angico), Cedrela fissilis (cedro) and Pelthophorum dubium (canafistula). The fungi was applied at different times of the production of seedlings. The fungi action was different between species and between the variables. Cedro was less responsive to application of Trichoderma spp. at any time of the seedlings production and the association of Trichoderma spp. With angico as the seed treatment favored the incidence of some pathogens, however the application later favored only the leaves development. In canafístula, the seed treatment provided control of pathogens associated and stimulated the emergence and production of leaves in seedlings, have potential to be used for seed treatment of this species.

Os objetivos deste trabalho foram: avaliar a qualidade fisiológica de sementes de cedro (Cedrela fissilis), procedentes de localidades do sul do Brasil, através de testes de vigor e avaliar diferentes tratamentos nas sementes para controle de patógenos e para promoção da germinação da espécie. Para tanto, foram utilizadas seis amostras com procedências dos Estados do Rio Grande do Sul, Santa Catarina e Paraná, as quais foram submetidas à determinação de teor de água, teste de germinação e primeira contagem, envelhecimento acelerado (testando períodos de exposição às condições de envelhecimento), emergência em viveiro e tratamento de sementes. Os tratamentos utilizados para controle de patógenos foram: testemunha (T0); físico com calor seco a 70°C±3°C por 48 horas (T1); extrato aquoso de alho (Allium sativum) (T2); biológico à base de Trichoderma spp. - Agrotich Plus® (T3); e químico com fungicida protetor Captan (T4). A germinação variou de 56 a 87%; o período de 48 h sob temperatura de 41 °C foi o mais eficiente para estratificar as amostras em níveis de vigor; a emergência variou de 51 a 88% e as variáveis de desempenho de plântulas analisadas conseguiram estratificar as amostras em níveis de vigor. Quanto ao tratamento de sementes, o calor seco e o tratamento à base de extrato de alho se mostram eficientes no controle de microrganismos em semente de cedro, sem prejuízos ao vigor destas.

The objectives of this study were to evaluate the physiological quality of Cedrela fissilis seeds, from different origins, using different vigor tests and evaluating different seeds treatments for control of pathogens and to promote the germination of the species. Therefore, six samples from different places in states of Rio Grande do Sul, Santa Catarina and Paraná were used, which were subjected to determination of water content, germination and first count of germination, accelerated aging (testing different periods), emergence, with assessments seedlings and seed treatment. The treatments were the following: control (T0); physical treatment with dry heat at 70°C±3°C for 48 hours (T1); aqueous extract of Allium sativum (T2); biological with product based on Trichoderma spp. - Agrotich Plus® (T3); and chemic with fungicide Captan (T4). Germination ranged 56-87%, the period of 48 hours at a temperature of 41 ° C has been more efficient to stratify the samples at levels of vigor; emergence ranged 51-88% and the performance of seedlings variables were able to stratify the samples analyzed at levels of vigor. Regarding seed treatments, dry heat and treatment based on extracts of A. sativum are effective in controlling micro-organisms in C. fissilis seeds, without damage to the vigor of them.

Este trabalho teve como objetivo testar os efeitos in vitro e in vivo de bioprotetores à base de Trichoderma spp. no controle do fungo Cylindrocladium candelabrum Viegas. Os testes in vitro (confronto direto e inoculação em folhas destacadas) foram compostos pelos seguintes tratamentos: T1 - somente C. candelabrum; T2 - isolado 06006S x C. candelabrum; T3 - isolado 53RR x C. candelabrum; T4 - isolado 5D x C. candelabrum; T5 - Agrotrich® x C. candelabrum; e T6 - Trichodel® x C. candelabrum. Todos os tratamentos foram eficientes inibindo o crescimento do fungo C. candelabrum em confrontação direta, e os isolados de Trichoderma spp. 53RR e 06006S, bem como o produto comercial Trichodel®, controlaram a mancha-foliar em folhas destacadas. Para complementar os testes in vitro, os produtos comerciais Agrotrich® e Trichodel® foram testados em mudas de E. saligna cultivadas em casa de vegetação, com os seguintes tratamentos: T1 - Testemunha: sem inoculação; T2 - inoculação de C. candelabrum; T3 - inoculação de C. candelabrum x Agrotrich®; T4 - inoculação de C. candelabrum x Trichodel®; T5 - somente Agrotrich®; e T6 - somente Trichodel®. Este produto apresentou os melhores resultados na redução dos danos causados pelo patógeno em mudas de E. saligna.

This study tested the effects of commercial biocontrol products on in vitro and in vivo trials against the fungal pathogen Cylindrocladium candelabrum Viegas, leaf blight agent on Eucalyptus saligna. The in vitro tests (direct confrontation and inoculation on detached leaves of E. saligna) were composed by the following treatments: T1 - only C. candelabrum, T2 - 06006S isolate x C. candelabrum, T3 - 53RR isolate x C. candelabrum, T4 - 5D isolate x C. candelabrum; T5 -Agrotrich® x C. candelabrum and T6 - Trichodel® x C. candelabrum. All isolates were efficient in controlling the growth of the pathogen during the direct confrontation test; similarly, Trichoderma spp. 53RR and 06006S, as well as the commercial product Trichodel ®, controlled the symptoms on the detached leaves. In order to complement the in vitro tests, the commercial products Agrotrich® and Trichodel® were tested on E. saligna seedlings cultivated in greenhouse. The following treatments were applied: T1 - control - plants without inoculation; T2 - plants inoculated with C. candelabrum; T3 - plants inoculated with C. candelabrum x Agrotrich®; T4 - plants inoculated with C. candelabrum x Trichodel®; T5 - plants inoculated with Agrotrich® and T6 - plants inoculated with Trichodel®. Trichodel® reduced C. candelabrum damage on E. saligna seedlings.

RESUMO A erva-mate (Ilex paraguariensis A. St.-Hil.) é uma espécie amplamente cultivada no sul do Brasil e países vizinhos. Suas folhas e galhos finos, preparados segundo método apropriado, fornecem o mate ou chimarrão, o mais popular chá consumido no Brasil e exportado para todo o mundo. Com o aumento da área plantada, sobretudo na última década, os problemas fitossanitários causados por pragas e doenças aumentaram e, dentre estes, a podridão-de-raízes vem se tornando um dos principais. A doença é causada por Fusarium spp. e provoca sérios danos decorrentes da destruição do sistema radicular, ocasionando, nos casos mais graves, morte da planta. A erva-mate é uma espécie que habita o sub-bosque da mata e o cultivo de forma homogênea a pleno sol pode ser um fator estressante ocasionando menor crescimento e desenvolvimento e também torná-la mais susceptível à podridão-de-raízes. Assim, foram instalados experimentos na área do Viveiro Florestal pertencente ao Departamento de Ciências Florestais da Universidade Federal de Santa Maria com o objetivo de confirmar tais hipóteses. O experimento foi desenvolvido em um esquema fatorial (2 x 5), composto pela inoculação de Fusarium spp. e por níveis de sombreamento. Foi observado que plantas submetidas aos menores níveis de sombreamento ou a pleno sol tiveram seu desenvolvimento prejudicado e ficaram mais suscetíveis à doença. As plantas inoculadas com Fusarium spp. tiveram menor desenvolvimento em comparação com as não inoculadas, especialmente nos menores níveis de sombreamento.

ABSTRACT The erva-mate (Ilex paraguariensis A. St.-Hil.) is a species broadly cultivated in southern Brazil and neighboring countries, whose leaves and fine branches, prepared according to the appropriate method, make the mate or “chimarrão”, the most popular tea consumed in Brazil and exported around the world. With an increasing planted area, mainly in the last decade, phytosanitary problems caused by pests and disease have increased in this crop and root-rot is becoming one of the main problems. The disease is caused by Fusarium spp. and it causes serious damage to the roots, in the most serious cases, leading to the death of the plant. The erva-mate is a species that inhabits the forest underwood, and cultivation in a homogeneous way to full sun can be a stressful factor for the species causing less growth and development and, also, more susceptibility to root-rot. Therefore, experiments were conducted in the Forest Nursery area belonging to Forest Sciences Department of the Federal University Santa Maria with the objective of confirming these assumptions. The experiment was conducted in a factorial design (2 x 5), composed by Fusarium spp. inoculation and by shading levels. It was verified that plants submitted to lower shading levels or full sun had less development and were more susceptible to the disease. The plants inoculated with Fusarium spp. had less development in comparison with non-inoculated plants, mainly at the lower shading levels.

RESUMO A erva-mate (Ilex paraguariensis A. St.-Hil.) é uma espécie amplamente cultivada na Região Sul do Brasil. Com o aumento da área plantada, sobretudo na última década, os problemas fitossanitários causados por pragas e doenças aumentaram e, dentre estes, a podridão-de-raízes vem se tornando um dos principais. Dentre os patógenos causadores dessa doença, Fusarium oxysporum Schlecht. e Fusarium solani (Mart.) Sacc são os principais. Esses fungos provocam sérios danos decorrentes da destruição do sistema radicular, ocasionando, nos casos mais graves, a morte da planta. Suspeita-se que a incidência e a severidade da doença são influenciadas pelo cultivo de ervais em diferentes níveis de sombreamento ou a pleno sol. Assim, foram instalados experimentos na área do Viveiro Florestal pertencente ao Departamento de Ciências Florestais/CCR/UFSM com o objetivo de confirmar tal hipótese. O experimento foi conduzido em um esquema fatorial (2 x 5), composto pela inoculação de Fusarium oxysporum e Fusarium solani e por níveis de sombreamento. Verificou-se que as plantas submetidas aos menores níveis de sombreamento ou a pleno sol ficaram predispostas à doença.

ABSTRACT Ilex paraguariensis A. St.-Hil. is a broadly cultivated species in the South of Brazil. With the increasing planted area, mainly in the last decade, fitosanitary problems caused by pest and diseases have increased in this crop, and the root-rot is becoming one of the main problems. Among the pathogens, Fusarium oxysporum Schlecht. and Fusarium solani (Mart.) Sacc are the main ones. These fungi provoke severe damages resulting in the destruction of the root system and, in the most serious cases, they cause the death of the plant. It is suspected that the incidence and severity of disease are influenced by cultivation of Ilex paraguariensis in different levels of shading or full sun. Therefore, experiments were conducted in the Forest Nursery area belonging to Forest Sciences Department/CCR/UFSM with the objective of confirming this assumption. The experiment was conducted in a factorial design (2 x 5), composed by Fusarium oxysporum and Fusarium solani inoculation and by shading levels. It was verified that the plants submitted to lower shading levels or full sun had their predisposed to the disease.