Chemical treatment of papaya seeds aiming at long-term storage and control of damping off

Damping off is a nursery disease of great economic importance in papaya and seed treatment may be an effective measure to control. The aim of this work was to evaluate the quality of papaya seeds treated with fungicides and stored under two environmental and packaging conditions. Additionally, the efficiency of fungicide treatments in the control of damping-off caused by Rhizoctonia solani was evaluated. Papaya seeds were treated with the fungicides Captan, Tolylfluanid and the mixture Tolylfluanid + Captan (all commercial wettable powder formulations). Seeds of the control group were not treated. The seeds were stored for nine months in two conditions: packed in aluminum coated paper and kept at 7 ± 1ºC and in permeable kraft paper and kept in non-controlled environment. At the beginning of the storage and every three months the seed quality (germination and vigor tests), emergence rate index, height, dry mass and damping of plants in pre and post-emergence (in contaminated substrate and mycelia-free substrate) were analyzed. Both storage conditions as well as the fungicide treatments preserved the germination and seed vigor. In the infested substrate, seedling emergence was favored by fungicides, but in post-emergence, fungicides alone did not control the damping off caused by R. solani. Symptoms of damping off were not observed in the clean substrate. The results showed that the fungicide treatments may be used to pretreat papaya seed for long-term storage and commercialization.

Ensaio sobre o controle de "damping-off' do cafeeiro

Testes de 3 fungicidas-Vapam, PCNB e Cupravit - foram conduzidos em 2 tipos de substrato, visando controlar "damping-off" do cafeeiro. Os substratos se comportaram de maneira diferente quanto ao número de plantas sadias obtidas. Os fungicidas agiram de maneira diferente em diferentes substratos. No substrato A os fungicidas diferiram na sua ação protetora enquanto que no solo B, não. No resultado finai, alguns tratamentos se mostraram superiores que outros.

Pyoluteorin production by Pseudomonas fluorescens strain CHA0 is involved in the suppression of Pythium damping-off of cress but not of cucumber

Pseudomonas fluorescens strain CHA0 is an effective biocontrol agent of various soilborne pathogens. It controls damping-off or root rot caused byPythium ultimum on cucumber, wheat and cress. Strain CHA0 synthesizes several antibiotic metabolites such as hydrogen cyanide, 2,4-diacetylphloroglucinol, and pyoluteorin. The role of pyoluteorin in the suppression of damping-off was investigated. Two Tn5 mutants (CHA660 and CHA661) of strain CHA0 were isolated which had lost the capacity to produce pyoluteorin but still produced 2,4-diacteylphloroglucinol and HCN. These mutants still inhibitedP. ultimum on malt agar (which favours the production of 2,4-diacetylphloroglucinol) but had partially lost the ability to inhibit this pathogen on King's B agar (which favours the production of pyoluteorin). The two pyoluteorin-negative mutants showed a reduced capacity to suppress damping-off of cress caused byP. ultimum but were as effective in the protection of cucumber against this pathogen as the wild-type strain. These results indicate that, depending on the plant, pyoluteorin production plays a role in the suppression of damping-off by strain CHA0 without being a major mechanism in disease suppression. We suggest that the contribution of pyoluteorin to the biocontrol activity of strain CHA0 is determined by the quantity of this antibiotic produced in the rhizosphere, which might depend on the root exudates of the host plant.

Essential oil of mustard to control Rhizoctonia solani causing seedling damping off and seedling blight in nursery

The essential oil extracted from mustard (Brassica rapa) seeds was evaluated for its effect on suppression of Rhizoctonia solani growth in vitro, and in field soils, for reducing saprophytic substrate colonization and seedling damping off and blight using snap beans as indicator plant, the in vitro growth was completely inhibited at a concentration of 50 mul/l. The saprophytic substrate colonization in soils 24 h after treatment was drastically reduced to 45% at 150 mul/kg soil concentration, in contrast to 100% colonization at concentrations of 0, 50, or 75 mul/kg. This recovery rate gradually declined to 6% and 60%, respectively, in nine days. A control of pre and post-emergence seedling damping off and blight in common beans (Phaseolus vulgaris), without any apparent phytotoxic effect was achieved by irrigating R. solani infested soils with water containing the emulsified essential oil to provide 150 mul/l soil volume ten days prior to planting, gave over 95%. The effect of the mustard essential oil was not influenced by the physical soil texture, and it appears to be a good substitute for methyl bromide fumigation in nurseries for seedling production.

Micorrización de Pinus greggii Engelm. como una medida de prevención del establecimiento de tres cepas de Fusarium oxysporum Schlecht. causantes de damping-off

Tesis (Maestría en Ciencias Forestales) U.A.N.L.

Involvement of phenazine-1-carboxylic acid, siderophores and hydrogen cyanide in suppression of Rhizoetonia solani and Pythium spp. damping-off by Pseudomonas oryzihabitans and Xenorhabdus nematophila

Entomopathogenic bacterial strains Pseudomonas (Flavimonas) oryzihabitans and Xenorhabdus nematophilus, both bacterial symbionts of the entomopathogenic nematodes Steinernema abbasi and S. carpocapsae have been recently used for suppression of soil-borne pathogens. Bacterial biocontrol agents (P. oryzihabitans and X nematophila) have been tested for production of secondary metabolites in vitro and their fungistatic effect,on mycelium and spore development of soil-borne pathogens. Isolates of Pythium spp. and Rhizoctonia solani, the causal agent of cotton damping-off, varied in sensitivity in vitro to the antibiotics phenazine-I-carboxylic acid (PCA), cyanide (HCN) and siderophores produced by bacterial strains shown previously to have potential for biological control of those pathogens. These findings affirm the role of the antibiotics PCA, HCN and siderophores in the biocontrol activity of these entomopathogenic strains and support earlier evidence that mechanisms of secondary metabolites are responsible for suppression of damping-off diseases. In the present studies colonies of R oryzihabitans showed production of PCA with presence of crystalline deposits after six days development and positive production where found as well in the siderophore's assay when X nematophila strain indicated HCN production in the in vitro assays. In vitro antifungal activity showed that bacteria densities of 101 to 10(6)cells/ml have antifungal activity in different media cultures. The results show further that isolates of Pythium spp. and R. solani insensitive to PCA, HCN and siderophores are present in the pathogen population and provide additional justification for the use of mixtures of entomopathogenic strains that employ different mechanisms of pathogen suppression to manage damping-off.

Biological control of Rhizoctonia solani Damping-off with a bacterium symbiotically associated with Steinernema abbasi

Rhizoctonia solani is a causal agent of damping-off of may cultivated plants. An isolate of the bacterium Pseudomonas oryzihabitans, symbiotically associated with the entomopathogenic nematode Steinernema abbasi, strongly inhibited the pathogen in vitro. The bacterium was firmly attached onto fungus mycelia and degraded the cell walls of the pathogen. In greenhouse experiments, bacterial suspension in sterile water applied in the soil, effectively controlled damping-off of radish.

Interação de fatores bióticos e abióticos na ocorrência de Damping-off em milho e feijoeiro

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Rhizoctonia solani as causative agent of damping off of Swiss chard in Europe

During September 2011, post-emergence damping off of Swiss chard (Beta vulgaris subsp. cicla L.) was observed in a greenhouse in Villa del Prado (Spain). About 20% of the seedlings showed damping off symptoms. Lesions were initially water soaked, dark brown necrosis of crown tissue, irregular in shape and sunken in appearance on large plants, causing the infected seedlings to collapse and eventually die. Rhizoctonia solani was isolated consistently from symptomatic plants. After morphological and molecular identification of the isolates, pathogenicity was tested by placing agar plugs of four isolates adjacent to the stem at the three or four true leaf stage. In inoculated plants, brown crown and stem necrosis occurred while control plants did not show disease symptoms. Pathogenicity using non-germinated seeds was also tested. All four isolates produced extensive damping off when inoculated on non-germinated seeds. To our knowledge, this is the first report of damping off of Swiss chard caused by R. solani in Europe.

Characterization of lipopeptides from Paenibacillus sp (IIRAC30) suppressing Rhizoctonia solani

The main aim was to identify the active compound against Rhizoctonia solani produced by the cassava endophyte Paenibacillus sp. IIRAC-30. The compounds produced were extracted with ethyl acetate and purified by Sephadex column prior to analysis by Q-TOF mass spectrometry. A C(15)-lipopeptide with an estimated molecular weight of 1036 Da and homologues were identified. The lipopeptide had a cyclic structure, which was deduced by interpreting the ESI-MS/MS spectra of main protonated homologues containing 15:0 FA, and the amino acid composition was Glu-Leu-Leu-Val-Asp-Leu-Leu. Therefore, the lipopeptides produced by isolate IIRAC-30 was characterized as a surfactin series. Thus, the main mechanism used by Paenibacillus sp. IIRAC-30 to suppress R. solani was elucidated. Furthermore, because lipopeptides active against phytopathogens generally show low toxicity to humans and the environment, the positive findings presented here suggest that the isolate IIRAC-30 could be a possible candidate for biocontrol of R. solani.

Bacterial subfamily of LuxR regulators that respond to plant compounds.

Pseudomonas fluorescens are rhizobacteria known for their biocontrol properties. Several antimicrobial functions are crucial for this process, and the experiments described here investigate the modulation of their expression during the plant-bacterium interaction. The role of a LuxR family regulator in interkingdom signaling has been investigated using genome-scale transcriptome analysis, gene promoter studies in vivo and in vitro, biocontrol assays, and response to plant compounds. PsoR, a LuxR solo or orphan regulator of P. fluorescens, was identified. PsoR is solubilized and activates a lux-box-containing promoter only in the presence of macerated plants, suggesting the presence of a plant molecule(s) that most likely binds to PsoR. Gene expression profiles revealed that genes involved in the inhibition of plant pathogens were affected by PsoR, including a chitinase gene, iron metabolism genes, and biosynthetic genes of antifungal compounds. 2,4-Diacetylphloroglucinol production is PsoR dependent both in vitro and in vivo. psoR mutants were significantly reduced for their ability to protect wheat plants from root rot, and damping-off caused by Pythium ultimum infection. PsoR most likely senses a molecule(s) in the plant and modulates expression of genes that have a role in biocontrol. PsoR and related proteins form a subfamily of LuxR family regulators in plant-associated bacteria.

The global regulator GacA of Pseudomonas fluorescens CHA0 is required for the suppression of root diseases in dicotyledons but not in Gramineae

Pseudomonas fluorescens strain CHA0 suppresses various plant diseases caused by soil-borne fungi. The pseudomonad produces the antimicrobial metabolites 2,4-diacetylphloroglucinol (Phl), pyoluteorin (Plt) and hydrogen cyanide, which are important for disease suppression, as well as the siderophores pyoverdine (Pvd), salicylic acid (Sal) and pyochelin (Pch). In the current work, a derivative of CHA0 with a mutation in the global regulator gene gacA (GacA−), which is unable to produce Phl, Plt and HCN, failed to protect the dicotyledonous plants cress and cucumber against damping-off caused by Pythium ultimum. In contrast, the GacA− mutant could still protect the Gramineae wheat and maize against damping-off mediated by the same strain of P. ultimum, and wheat against take-all caused by Gaeumannomyces graminis. However, the GacA− mutant overproduced Pch and Pvd. To gain more insight into disease protection afforded by the GacA− mutant, a GacA− Pvd− double mutant (strain CHA496) was constructed by gene replacement. Strain CHA496 overproduced Pch and Sal compared with CHA0 and protected wheat against P. ultimum and G. graminis, whereas cress and cucumber were not protected. Addition of FeCl3 repressed Pch and Sal production by strain CHA496 in vitro and impaired the protection of wheat in soil microcosms. In conclusion, a functional gacA gene was necessary for the protection of dicotyledons against root diseases, but not for that of Gramineae. Results indicated also that Pch and/or Sal were involved in the ability of the GacA− Pvd− mutant of CHA0 to suppress root diseases in Gramineae.

Efeito do tratamento de sementes de algodão com fungicidas no controle do tombamento de plântulas causado por Rhizoctonia solani

Este trabalho foi desenvolvido na Embrapa Agropecuária Oeste, em Dourados, MS, e teve por objetivo avaliar a eficiência de alguns fungicidas, aplicados no tratamento de sementes de algodão (Gossypium hirsutum), no controle do tombamento de plântulas causado por Rhizoctonia solani. Foi realizado teste em casa de vegetação, utilizando a cultivar DeltaOpal. Sementes tratadas e não tratadas com fungicidas foram semeadas em areia contida em bandejas plásticas, dispostas em orifícios individuais, eqüidistantes e a 3 cm de profundidade. A inoculação com R. solani foi feita pela distribuição homogênea do inóculo do fungo na superfície do substrato. O fungo foi cultivado por 35 dias em sementes de aveia autoclavadas e trituradas em moinho (1 mm). Foram utilizados 9 g de inóculo por bandeja de areia. Foi observado efeito do tratamento fungicida na emergência inicial e final de plântulas, com destaque para triadimenol + pencycuron + tolylfluanid e triadimenol + tolylfluanid, seguidos de carboxin + thiram, triadimenol e carboxin + thiram + carbendazim. Os tratamentos mais eficientes no controle do tombamento de pós-emergência do algodoeiro foi obtido com a mistura triadimenol + pencycuron + tolylfluanid, seguida de triadimenol, triadimenol + tolylfluanid e carboxin+thiram. Nenhum dos fungicidas testados foi fitotóxico ao algodão.

Seleção de plantas resistentes e de fungicidas para o controle da podridão do colo do maracujazeiro causada por Nectria haematococca

A podridão do colo do maracujazeiro (Passiflora edulis f. flavicarpa), causada por Nectria haematococca e Phytophthora spp., é um dos principais problemas da cultura no Brasil, sendo responsável pelo decréscimo da produtividade e constantes migrações da cultura. O controle da doença é basicamente preventivo, visando a não introdução dos patógenos na área. O presente trabalho visou 1) avaliar métodos de inoculação de N. haematococca e a suscetibilidade do maracujazeiro amarelo ao patógeno em diferentes idades; 2) a ocorrência de "damping-off"; 3) o comportamento de diferentes espécies do gênero Passiflora e de genótipos de P. edulis f. flavicarpa ao patógeno; e 4) realizar testes de controle químico. Inoculações no colo de maracujazeiro amarelo proporcionaram maiores níveis de doença comparadas às inoculações no sistema radicular. Os resultados sugerem que N. haematococca seja um patógeno que penetra por ferimentos. A mortalidade foi maior quando a inoculação foi realizada em plantas mais jovens e quando N. haematococca e Phytophthora nicotianae estavam associados. Dentre as 17 espécies do gênero Passiflora avaliadas para resistência a N. haematococca, P. nitida, P. laurifolia e P. alata foram as menos lesionadas. Os genótipos de P. edulis f. flavicarpa mais resistentes ao patógeno foram os procedentes de Morretes (PR), Sapucaí (SP) e a cultivar Maguari. Os fungicidas prochloraz, thiabendazole, thiram+thiabendazole, carbendazim, triflumizole e captan exerceram controle erradicante em solo infestado com N. haematococca. Os fungicidas prochloraz e carbendazim evitaram a morte de plantas em tratamento curativo, com os melhores resultados quando aplicados dois dias após a inoculação, comparado a sete dias.