A biostimulant yeast, Hanseniaspora opuntiae, modifies Arabidopsis thaliana root architecture and improves the plant defense response against Botrytis cinerea.

MAIN CONCLUSION: The biostimulant Hanseniaspora opuntiae regulates Arabidopsis thaliana root development and resistance to Botrytis cinerea. Beneficial microbes can increase plant nutrient accessibility and uptake, promote abiotic stress tolerance, and enhance disease resistance, while pathogenic microorganisms cause plant disease, affecting cellular homeostasis and leading to cell death in the most critical cases. Commonly, plants use specialized pattern recognition receptors to perceive beneficial or pathogen microorganisms. Although bacteria have been the most studied plant-associated beneficial microbes, the analysis of yeasts is receiving less attention. This study assessed the role of Hanseniaspora opuntiae, a fermentative yeast isolated from cacao musts, during Arabidopsis thaliana growth, development, and defense response to fungal pathogens. We evaluated the A. thaliana-H. opuntiae interaction using direct and indirect in vitro systems. Arabidopsis growth was significantly increased seven days post-inoculation with H. opuntiae during indirect interaction. Moreover, we observed that H. opuntiae cells had a strong auxin-like effect in A. thaliana root development during in vitro interaction. We show that 3-methyl-1-butanol and ethanol are the main volatile compounds produced by H. opuntiae. Subsequently, it was determined that A. thaliana plants inoculated with H. opuntiae have a long-lasting and systemic effect against Botrytis cinerea infection, but independently of auxin, ethylene, salicylic acid, or jasmonic acid pathways. Our results demonstrate that H. opuntiae is an important biostimulant that acts by regulating plant development and pathogen resistance through different hormone-related responses.

Gadolinium Protects Arabidopsis thaliana against Botrytis cinerea through the Activation of JA/ET-Induced Defense Responses.

Plant food production is severely affected by fungi; to cope with this problem, farmers use synthetic fungicides. However, the need to reduce fungicide application has led to a search for alternatives, such as biostimulants. Rare-earth elements (REEs) are widely used as biostimulants, but their mode of action and their potential as an alternative to synthetic fungicides have not been fully studied. Here, the biostimulant effect of gadolinium (Gd) is explored using the plant-pathosystem Arabidopsis thaliana-Botrytis cinerea. We determine that Gd induces local, systemic, and long-lasting plant defense responses to B. cinerea, without affecting fungal development. The physiological changes induced by Gd have been related to its structural resemblance to calcium. However, our results show that the calcium-induced defense response is not sufficient to protect plants against B. cinerea, compared to Gd. Furthermore, a genome-wide transcriptomic analysis shows that Gd induces plant defenses and modifies early and late defense responses. However, the resistance to B. cinerea is dependent on JA/ET-induced responses. These data support the conclusion that Gd can be used as a biocontrol agent for B. cinerea. These results are a valuable tool to uncover the molecular mechanisms induced by REEs.

Reproducibility of the Development and Validation Process of Standard Area Diagram by Two Laboratories: An Example Using the Botrytis cinerea/Gerbera jamesonii Pathosystem.

Standard area diagrams (SADs) are plant disease severity assessment aids demonstrated to improve the accuracy and reliability of visual estimates of severity. Knowledge of the sources of variation, including those specific to a lab such as raters, specific procedures followed including instruction, image analysis software, image viewing time, etc., that affect the outcome of development and validation of SADs can help improve standard operating practice of these assessment aids. As reproducibility has not previously been explored in development of SADs, we aimed to explore the overarching question of whether the lab in which the measurement and validation of a SAD was performed affected the outcome of the process. Two different labs (Lab 1 and Lab 2) measured severity on the individual diagrams in a SAD and validated them independently for severity of gray mold (caused by Botrytis cinerea) on Gerbera daisy. Severity measurements of the 30 test images were performed independently at the two labs as well. A different group of 18 raters at each lab assessed the test images first without, and secondly with SADs under independent instruction at both Lab 1 and 2. Results showed that actual severity on the SADs as measured at each lab varied by up to 5.18%. Furthermore, measurement of the test image actual values varied from 0 to up to 24.29%, depending on image. Whereas at Lab 1 an equivalence test indicated no significant improvement in any measure of agreement with use of the SADs, at Lab 2, scale shift, generalized bias, and agreement were significantly improved with use of the SADs (P ≤ 0.05). An analysis of variance indicated differences existed between labs, use of the SADs aid, and the interaction, depending on the agreement statistic. Based on an equivalence test, the interrater reliability was significantly (P ≤ 0.05) improved at both Lab 1 and Lab 2 as a result of using SADs as an aid to severity estimation. Gain in measures of agreement and reliability tended to be greatest for the least able raters at both Lab 1 and Lab 2. Absolute error was reduced at both labs when raters used SADs. The results confirm that SADs are a useful tool, but the results demonstrated that aspects of the development and validation process in different labs may affect the outcome.

Economic Viability of Tomato Cultivation in Organic Farming System

ABSTRACT The aim of the present study is to evaluate the economic viability of tomato cultivation in a system of organic farming, and to compare it with the conventional farming system. The fixed and variable costs of both cultivation systems were calculated, as well as the costs of disease control with the application of alternative products and pesticides. The revenues were computed using commercial production and the direct sales price. The total cost per area was higher in the conventional system, while the cost per plant was greater in the organic system, since it used a lower plant population density. In the conventional system, 2.33 times more was spent on plant management, due to the fact that the number of sprayings was greater than that carried out in organic farming. We further find that companion planting with other species such as coriander, in addition to helping with the management of the main crop, is important for economic sustainability as it is an extra source of income. Considering the specificities of the two systems, the profitability of organic cultivation, even accounting for its lower productivity , could exceed that of conventional cultivation.

Efeito de extratos de cogumelos na indução de fitoalexinas e no controle de oídio da soja em casa de vegetação / Mushrooms extracts effect on the induction of phytoalexins and on the control of soybean powdery mildew in greenhouse

O oídio (Erysiphe diffusa) tornou-se uma das principais doenças que afeta a cultura da soja (Glycine max), podendo causar redução significativa no rendimento da cultura. Esse trabalho teve por objetivo avaliar o efeito dos extratos aquosos de Agaricus blazei, Lentinula edodes e Pycnoporus sanguineus no controle do oídio em casa-devegetação e na indução de fitoalexinas em cotilédones de soja. O delineamento experimental utilizado foi inteiramente casualizado com cinco repetições. Como controle utilizou-se fungicida (enxofre ­ 3g/L) e testemunha - sem aplicação. Os tratamentos utilizados foram: extratos aquosos de Agaricus blazei, Lentinula edodes e Pycnoporus sanguineus na concentração de 20 %v/v sendo estes aplicados aos 61 e 68 dias após a semeadura. Avaliou-se a incidência e severidade aos 59, 65, 69, 73 e 77 dias após a semeadura em todos os folíolos de cada planta. Com os dados da severidade, calculouse a área abaixo da curva de progresso da doença (AACPD). Para a incidência, detectou-se diferença significativa entre o fungicida e os demais tratamentos. Para severidade e AACPD, observou-se que os extratos diferiram da testemunha, sendo que L. edodes mostrou ser mais eficiente que os demais tratamentos. Para o bioensaio de fitoalexinas foi aplicado sobre cada cotilédone de soja uma alíquota de 75 µL de cada extrato aquoso nas concentrações de 1%, 2%, 4% e 10%. Após 20 horas, os mesmos foram transferidos para erlenmeyers contendo 15 mL de água destilada esterilizada e deixados em agitação por 1 h para extração da fitoalexina formada. A absorbância foi determinada a 285 nm em espectrofotômetro. Observou-se um maior acúmulo de fitoalexina nas maiores concentrações dos extratos aquosos.

The powdery mildew (Erysiphe diffusa) became one of the main diseases that effects soybean (Glycine max) crops. The aim was to evaluate the effect of Agaricus blazei, Lentinula edodes and Pycnoporus sanguineus aquous extracts on the control of powdery mildew in greenhouse and on the induction of phytoalexins in soybean cotyledons. The control treatments consisted of fungicide (Sulphur ­ 3 g/L) and control without application. The treatments were applied at 61 and 68 days after sowing (DAS). The incidence and severity was evaluated at 59, 65, 69, 73 and 77 DAS in all the folioles of each plant. With the severity values, the area under the disease progress curve (AUDPC) was estimated. To the incidence, a significative difference was detected between the fungicide and the other treatments. For severity and the AUDPC, the extracts and the fungicide differed from the control and L. edodes was the most efficient treatment. For the phytoalexins production was applied 75 mL to each soybean cotyledon of each aqueous extract at 1%, 2%, 4% and 10% concentrations. After 20 hours, they were transferred to erlenmayer flask containing 15 mL of sterile distilled water and remained stirring for 1 h for extraction of phytoalexin. The absorbance was measured at 285 nm in a spectrophotometer. The highest accumulation of the phytoalexins was observed on highests concentrations of the aqueous extracts.

Relações de causa e efeito entre condições ambientais, urediniósporos e severidade de ferrugem-asiática da soja / Relations of cause and effect among environmental conditions, airborne urediniospores and asian soybean rust severity

O objetivo deste trabalho foi estabelecer relações entre condições ambientais, o número de urediniósporos de Phakopsora pachyrhizi e a severidade da ferrugem-asiática-da-soja. Para quantificar urediniósporos de P. pachyrhizi, coletores foram instalados em campos de soja, em Itambé e Mandaguari, PR. Avaliações de severidade foram realizadas e dados climáticos foram coletados nos dois locais. As variáveis elaboradas foram: número de dias com umidade favorável (>60%), número de dias com temperatura favorável (18 a 32º C), preciptação acumulada e urediniósporos coletados. Uma variável baseada no controle químico também foi elaborada. Correlações entre as variáveis foram estimadas. A análise de trilha foi empregada para desdobrar as correlações. Cerca de 76% da variação das relações de causa e efeito entre as variáveis foi explicada pela análise de trilha. O número de dias para efetuar o controle químico teve efeito direto sob a severidade. O número de dias com temperatura favorável e as precipitações tiveram efeito indireto na severidade, atuando via manutenção de condições favoráveis de umidade.

The objective of this work was to study the relationships among environmental conditions, airborne urediniospores of P. pachyrhizi and the Asian soybean rust (ASR) severity. To quantify airbone urediniospores, collectors were installed in soybean fields in Itambé and Mandaguari, PR. Severity assessments were conducted and climatic data were collected in the two locations. The variables elaborated were: number of days with favorable humidity (<60%), number of days with favorable temperature (18-32ºC), rainfall accumulated and urediniospores collected. A variable based on the chemical control was also created. Correlations between variables were estimated. Path analysis was performed for correlations. Path analysis explained about 76% of the relations of cause and effect among the variables. Number of days to the chemical control had direct effect on severity. Number of days with favorable temperature and rainfall had indirect effect on severity. This two variables provided favorable humidity conditions for ASR development.

Atividades antibacteriana, antifúngica e indutora de fitoalexinas de hidrolatos de plantas medicinais / Antibacterial, antifungal and phytoalexins induction activities of hydrolates of medicinal plants

(O objetivo do trabalho foi avaliar a atividade como antifúngico, antibacteriano e indutor da produção de fitoalexinas dos hidrolatos de Helietta apiculata (canela-de-veado) (HA), Conyza canadensis (buva) (CC) e Cymbopogon nardus (citronela) (CN) nas concentrações de 1, 5, 10, 15, 20 e 25%, buscando seu uso no controle alternativo de doenças em plantas. Para o ensaio de fitoalexinas foram utilizados mesocótilos estiolados de sorgo. Para o efeito antibacteriano foi avaliado o crescimento da bactéria Xanthomonas campestris pv. campestris em meio caldo nutriente suplementado com os tratamentos, tendo como testemunha mistura de antibióticos (22,5 mg/L de oxitetraciclina + 225 mg/L de estreptomicina). O efeito antifúngico foi avaliado através da mensuração do crescimento vegetativo, esporulação, germinação de esporos e desenvolvimento do tubo germinativo de Alternaria brassicae, sendo o fungicida azoxystrobin (0,08 g i.a./L) usado como controle. Foi observado acréscimo na síntese de fitoalexinas com o aumento na concentração dos hidrolatos, sendo o melhor resultado obtido com CN, o qual promoveu aumento de cerca de 4,3 vezes mais, seguido de HA (2,5 vezes) e CC (2,1 vezes), em relação a testemunha água. Resultados semelhantes foram obtidos para crescimento bacteriano com os seguintes resultados de inibição (em %) no desenvolvimento da bactéria: CN: 29,8, HA: 14,9, CC: 14,6 e antibióticos: 97,7. Com relação à ação antifúngica foi observada inibição do desenvolvimento dos tubos germinativos, sendo que os hidrolatos de CC e HA inibiram 69,2 e 56,2%, respectivamente, resultado semelhante ao fungicida. O hidrolato de CN não apresentou efeito antifúngico. Estes resultados indicam a presença de compostos indutores de fitoalexinas, antibacterianos e antifúngicos nesses hidrolatos, porém em baixas concentrações.

The aim of this work was to evaluate the antifungical, antibacterial and phytoalexins elicitors activities ofhydrolates, for using in the alternative control of plant diseases. The hydrolates of Helietta apiculata(HA), Conyza canadensis (CC) and Cymbopogon nardus (CN) were used in the concentrations of 1, 5,10, 15, 20 and 25%. In the phytoalexins assay, sorghum etiolated mesocotyls were used. The antibacterialeffect was evaluated on the growth of the bacterium Xanthomonas campestris pv. campestris in liquidnutrient medium and antibiotic (oxytetraciclin 22.5 mg/L + streptomycin 225 mg/L) was used as controltreatment. The antifungical effect was evaluated on the micelial growth, esporulation, conidia germinationand development of germinative tubes of Alternaria brassicae. The fungicide azoxystrobin (0.08 g a.i./L) was used as control treatment. There was increment in the phytoalexins synthesis with the increase inhydrolates concentration. The better result was obtained with CN that promoted increase 4.3 times largerin relation to the value of the control treatment with water, followed by HA (2.5 times) and CC (2.1 times).Similar results were obtained to the antibacterial activity with the following inhibition results (%) in the development of the bacterium: CN: 29.8, HA: 14.9, CC: 14.6 and antibiotic: 97.7. The main antifungicaleffect was observed on the development of the germinative tubes. Hydrolates of CC and HA showedinhibition of up to 69.2 and 56.2%, respectively, being similar to the fungicide azoxystrobin. The hydrolateof CN did not show antifungical effect. These results indicate the presence of phytoalexins elicitors, and antibacterial and antifungical compounds in those hydrolates, however in low concentrations

Efeito do extrato aquoso de cúrcuma (curcuma longa) em Xanthomonas axonopodis pv. manihotis / Effect of aqueous extract from turmeric (Curcuma longa) on Xanthomonas axonopodis pv. manihotis

Avaliou-se o controle in vitro de Xanthomonas axonopodis pv. manihotis mediante o uso de extrato aquoso de quatro genótipos de cúrcuma provenientes de cultivos de Jaboticabal-SP, Mara Rosa-GO, Maringá-PR e Mercedes-PR, bem como o efeito curativo, através do tratamento de manivas de mandioca infectadas com o patógeno e plantio em condições de campo. No experimento in vitro, o extrato de cúrcuma causou inibição total do crescimento da bactéria, na concentração de 10, para o material proveniente de Mercedes, enquanto que, para a cúrcuma de Jaboticabal, houve controle total a 15 e o de Mara Rosa a 20. A cúrcuma proveniente de Maringá não inibiu totalmente o crescimento, em nenhuma das concentrações utilizadas. No experimento in vivo, o brotamento das plantas foi pouco, devido ao grau de infecção das manivas. O extrato bruto de cúrcuma a 10 proveniente de Mercedes foi prejudicial para a mandioca em condição de campo, reduzindo o estande em relação aos tratamentos controle. Possivelmente, houve ação tóxica direta sobre a fisiologia da planta ou indução de suscetibilidade. Na concentração de 1 da cúrcuma proveniente de Maringá, não houve diferença estatística em relação ás testemunhas, para o parâmetro estande de plantas. O controle químico utilizado não foi eficiente, pois se comportou igual à testemunha água. Com relação à severidade e à produtividade não se conservaram diferenças significativas entre os tratamentos. Os resultados indicam que, embora haja atividade antibacteriana a X. axonopodis pv. manihotis, os extratos de cúrcuma, nas concentrações utilizadas, não apresentam efeito curativo em manivas de mandioca infectadas pelo patógeno

medicinalAbstractThe control of Xanthomonas axonopodis pv. manihotis was evaluated in vitro by using aqueous extractof four turmeric genotypes from Jaboticabal-SP, Mara Rosa-GO, Maringá-PR and Mercedes-PR, as wellas in vivo, by treatment of infected cassava stems and their cultivation at field conditions. The resultsshowed that in vitro experiment, turmeric extract inhibited completely the bacteria growth in theconcentration of 10% for the genotype from Mercedes, while for the Jaboticabal's turmeric there was a total control at 15% and for Mara Rosa at 20%. Turmeric genotype from Maringá did not show fullinhibition of the bacterial growth in none of the extract concentrations used. At field conditions, sproutingswere extremely low, due to the stems infection degree. Turmeric extract at 10%, from Mercedes, washarmful for the cassava, reducing stand regarding the control treatments. Possibly there was a directtoxic action on the plant physiology or susceptibility induction. But in the concentration of 1% ofturmeric from Maringá, there was no statistical difference in relation to the control treatment for the plantstand. The chemical control was not completely efficient. There was no statistical difference amongtreatments for both severity and productivity. The results indicate that, although presenting antibacterialactivity to X. axonopodis pv. manihotis, the turmeric extracts, in the used concentrations, did notpresent curative effect in cassava stems infected with the pathogen.Key words: Bacterial blight of cassava, alternative control, medicinal plant

Avaliação da eficácia da tintura etanólica de guaco (Mikania glomerata) no controle da podridão negra (Xanthomonas campestris pv. campestris) em couve-flor / Effect of alcoholic extract of guaco (Mikania glomerata) on the control of dark rot (Xanthomonas campestris pv. campestris) in cauliflower

Com a prática da irrigação e novos híbridos de couve-flor, é possível produzir durante todo o ano e com alta produtividade. Mas, a cultura tem sido afetada por doenças a exemplo da podridão negra causada por Xanthomonas campestris pv. campestris, fomentando novas pesquisas para seu controle. Com o objetivo de verificar o potencial de Mikania glomerata no controle dessa doença, a tintura etanólica 50 ºGL dessa planta medicinal foi avaliada quanto: atividade antimicrobiana in vitro através do crescimento bacteriano em tubos de ensaio contendo 100, 250, 500 e 1000 mg L-1 da tintura; indução de resistência local ou sistêmica em planta de couve-flor aos 25 dias de idade, em casa de vegetação, através da pulverização de tintura concomitantemente e três dias antes da inoculação com o patógeno, sendo água e calda bordaleza tratamentos controle; atividade de peroxidases em folhas tratadas e não tratadas de couve-flor, colhidas concomitantemente e as 24, 48, 72 h da pulverização da tintura e, após pulverização-inoculção. A tintura etanólica, in vitro, promoveu inibição no crescimento bacteriano, a partir da concentração de 250 mg L-1. Nas concentrações de 500 mg L-1 e 1000 mg L-1 foram observadas, respectivamente, 24 e 38 de inibição do crescimento bacteriano. Nas plantas de couve-flor foi observada redução da doença apenas em folhas tratadas com 100 e 500 mg L-1 de tintura, aplicada concomitantemente à inoculação, comportamento este semelhante ao da calda bordaleza, indicando que o controle através da tintura de guaco é através de atividade antimicrobiana direta. Ficou indicado que a indução de peroxidases ocorreu devido ao processo infeccioso e não em função dos tratamentos com tintura etanólica de guaco. Estes resultados indicam o potencial da tintura de guaco para o controle preventivo da podridão negra da couve-flor.

With the use of irrigation and new hybrids of cauliflower, it is possible to get production during all theyear with hight yield. However, the crop has been affected by diseases, as the dark rot caused by X.campestris pv. campestris. The objective of this research work was to study the potential of Mikaniaglomerata for the control of this disease. Alcoholic extract 50 ºGL of M. glomerata was evaluatedregarding to: in vitro antimicrobial activity through bacterial growth in 100, 250, 500 and 1000 mg L-1 ofthe alcoholic extract; induction of local or systemic resistance in 25 days old cauliflower, with spray ofalcoholic extract concomitantly and three days before the inoculation with the pathogen (water andbordeau mixture were used as control); peroxidases activity in leaves of cauliflower treated and nottreated, and harvested concomitantly, 24, 48 and 72 hours after spraying the alcoholic extract and alsoafter inoculation. The alcoholic extract of M. glomerata showed inhibition of the bacterial growth invitro at the concentrations of 250, 500 and 1000 mg L-1. The concentrations of 500 mg L-1 and 1000 mg L-1inhibited 24% and 38% of the bacterial growth. This inhibition could be due to antibacterial compoundsin the alcoholic extract. An inhibition of the disease in vivo occurred only in the leaves treated with 100and 500 mg L-1 of alcoholic extract when applied concomitantly with the bacteria. This result was similarto bordeau mixture, indicating a control by direct antimicrobial activity. There was no systemic resistenceinduction for all treatments. The peroxidases induction was due to infectious pathogen process and notto the treatments with alcoholic extract. The results indicate the potential of M. glomerata alcoholicextract for the preventive control of cauliflower dark rot disease