A disease predictive model based on epidemiological factors for the management of bacterial leaf blight of rice / Um modelo preditivo de doenças baseado em fatores epidemiológicos para o manejo da queima bacteriana das folhas do arroz

Rice is a widely consumed staple food for a large part of the world's human population. Approximately 90% of the world's rice is grown in Asian continent and constitutes a staple food for 2.7 billion people worldwide. Bacterial leaf blight (BLB) caused by Xanthomonas oryzae pv. oryzae is one of the devastating diseases of rice. A field experiment was conducted during the year 2016 and 2017 to investigate the influence of different meteorological parameters on BLB development as well as the computation of a predictive model to forecast the disease well ahead of its appearance in the field. The seasonal dataset of disease incidence and environmental factors was used to assess five rice varieties/ cultivars (Basmati-2000, KSK-434, KSK-133, Super Basmati, and IRRI-9). The accumulated effect of two year environmental data; maximum and minimum temperature, relative humidity, wind speed, and rainfall, was studied and correlated with disease incidence. Average temperature (maximum & minimum) showed a negative significant correlation with BLB disease and all other variables; relative humidity, rainfall, and wind speed had a positive correlation with BLB disease development on individual varieties. Stepwise regression analysis was performed to indicate potentially useful predictor variables and to rule out incompetent parameters. Environmental data from the growing seasons of July to October 2016 and 2017 revealed that, with the exception of the lowest temperature, all environmental factors contributed to disease development throughout the cropping season. A disease prediction multiple regression model was developed based on two-year data (Y = 214.3-3.691 Max T-0.508 Min T + 0.767 RH + 2.521 RF + 5.740 WS), which explained 95% variability. This disease prediction model will not only help farmers in early detection and timely management of bacterial leaf blight disease of rice but may also help reduce input costs and improve product quality and quantity. The model will be both farmer and environmentally friendly.

O arroz é um alimento básico amplamente consumido por grande parte da população humana mundial. Aproximadamente 90% do arroz do mundo é cultivado no continente asiático e constitui um alimento básico para 2,7 bilhões de pessoas em todo o mundo. O crestamento bacteriano das folhas (BLB) causado por Xanthomonas oryzae pv. oryzae é uma das doenças devastadoras do arroz. Um experimento de campo foi realizado durante os anos de 2016 e 2017 para investigar a influência de diferentes parâmetros meteorológicos no desenvolvimento do BLB, bem como o cálculo de um modelo preditivo para prever a doença bem antes de seu aparecimento em campo. O conjunto de dados sazonais de incidência de doenças e fatores ambientais foi usado para avaliar cinco variedades/cultivares de arroz (Basmati-2000, KSK-434, KSK-133, Super Basmati e IRRI-9). O efeito acumulado de dados ambientais de dois anos; temperatura máxima e mínima, umidade relativa do ar, velocidade do vento e precipitação pluviométrica foram estudados e correlacionados com a incidência da doença. A temperatura média (máxima e mínima) apresentou correlação significativa negativa com a doença BLB e todas as outras variáveis; umidade relativa, precipitação e velocidade do vento tiveram uma correlação positiva com o desenvolvimento da doença BLB em variedades individuais. A análise de regressão stepwise foi realizada para indicar variáveis preditoras potencialmente úteis e para descartar parâmetros incompetentes. Os dados ambientais das safras de julho a outubro de 2016 e 2017 revelaram que, com exceção da temperatura mais baixa, todos os fatores ambientais contribuíram para o desenvolvimento da doença ao longo da safra. Um modelo de regressão múltipla de previsão de doença foi desenvolvido com base em dados de dois anos (Y = 214,3-3,691 Max T-0,508 Min T + 0,767 RH + 2,521 RF + 5,740 WS), que explicou 95% de variabilidade. Este modelo de previsão de doenças não só ajudará os agricultores na detecção precoce e gestão atempada da doença bacteriana das folhas do arroz, mas também pode ajudar a reduzir os custos de insumos e melhorar a qualidade e a quantidade do produto. O modelo será agricultor e ambientalmente amigável.

A disease predictive model based on epidemiological factors for the management of bacterial leaf blight of rice

Abstract Rice is a widely consumed staple food for a large part of the worlds human population. Approximately 90% of the worlds rice is grown in Asian continent and constitutes a staple food for 2.7 billion people worldwide. Bacterial leaf blight (BLB) caused by Xanthomonas oryzae pv. oryzae is one of the devastating diseases of rice. A field experiment was conducted during the year 2016 and 2017 to investigate the influence of different meteorological parameters on BLB development as well as the computation of a predictive model to forecast the disease well ahead of its appearance in the field. The seasonal dataset of disease incidence and environmental factors was used to assess five rice varieties/ cultivars (Basmati-2000, KSK-434, KSK-133, Super Basmati, and IRRI-9). The accumulated effect of two year environmental data; maximum and minimum temperature, relative humidity, wind speed, and rainfall, was studied and correlated with disease incidence. Average temperature (maximum & minimum) showed a negative significant correlation with BLB disease and all other variables; relative humidity, rainfall, and wind speed had a positive correlation with BLB disease development on individual varieties. Stepwise regression analysis was performed to indicate potentially useful predictor variables and to rule out incompetent parameters. Environmental data from the growing seasons of July to October 2016 and 2017 revealed that, with the exception of the lowest temperature, all environmental factors contributed to disease development throughout the cropping season. A disease prediction multiple regression model was developed based on two-year data (Y = 214.3-3.691 Max T-0.508 Min T + 0.767 RH + 2.521 RF + 5.740 WS), which explained 95% variability. This disease prediction model will not only help farmers in early detection and timely management of bacterial leaf blight disease of rice but may also help reduce input costs and improve product quality and quantity. The model will be both farmer and environmentally friendly.

Resumo O arroz é um alimento básico amplamente consumido por grande parte da população humana mundial. Aproximadamente 90% do arroz do mundo é cultivado no continente asiático e constitui um alimento básico para 2,7 bilhões de pessoas em todo o mundo. O crestamento bacteriano das folhas (BLB) causado por Xanthomonas oryzae pv. oryzae é uma das doenças devastadoras do arroz. Um experimento de campo foi realizado durante os anos de 2016 e 2017 para investigar a influência de diferentes parâmetros meteorológicos no desenvolvimento do BLB, bem como o cálculo de um modelo preditivo para prever a doença bem antes de seu aparecimento em campo. O conjunto de dados sazonais de incidência de doenças e fatores ambientais foi usado para avaliar cinco variedades/cultivares de arroz (Basmati-2000, KSK-434, KSK-133, Super Basmati e IRRI-9). O efeito acumulado de dados ambientais de dois anos; temperatura máxima e mínima, umidade relativa do ar, velocidade do vento e precipitação pluviométrica foram estudados e correlacionados com a incidência da doença. A temperatura média (máxima e mínima) apresentou correlação significativa negativa com a doença BLB e todas as outras variáveis; umidade relativa, precipitação e velocidade do vento tiveram uma correlação positiva com o desenvolvimento da doença BLB em variedades individuais. A análise de regressão stepwise foi realizada para indicar variáveis preditoras potencialmente úteis e para descartar parâmetros incompetentes. Os dados ambientais das safras de julho a outubro de 2016 e 2017 revelaram que, com exceção da temperatura mais baixa, todos os fatores ambientais contribuíram para o desenvolvimento da doença ao longo da safra. Um modelo de regressão múltipla de previsão de doença foi desenvolvido com base em dados de dois anos (Y = 214,3-3,691 Max T-0,508 Min T + 0,767 RH + 2,521 RF + 5,740 WS), que explicou 95% de variabilidade. Este modelo de previsão de doenças não só ajudará os agricultores na detecção precoce e gestão atempada da doença bacteriana das folhas do arroz, mas também pode ajudar a reduzir os custos de insumos e melhorar a qualidade e a quantidade do produto. O modelo será agricultor e ambientalmente amigável.

Suppression of Xanthomonas oryzae pv. oryzae biofilm formation by Acacia mangium methanol leaf extract / Supressão de Xanthomonas oryzae pv. oryzae: formação do biofilme oryzae pelo extrato de folhas de metanol de Acacia mangium

Abstract Xanthomonas oryzae pv. oryzae (Xoo), a pathogen responsible for rice bacterial leaf blight, produces biofilm to protect viable Xoo cells from antimicrobial agents. A study was conducted to determine the potency of Acacia mangium methanol (AMMH) leaf extract as a Xoo biofilm inhibitor. Four concentrations (3.13, 6.25, 9.38, and 12.5 mg/mL) of AMMH leaf extract were tested for their ability to inhibit Xoo biofilm formation on a 96-well microtiter plate. The results showed that the negative controls had the highest O.D. values from other treatments, indicating the intense formation of biofilm. This was followed by the positive control (Streptomycin sulfate, 0.2 mg/mL) and AMMH leaf extract at concentration 3.13 mg/mL, which showed no significant differences in their O.D. values (1.96 and 1.57, respectively). All other treatments at concentrations of 6.25, 9.38, and 12.5 mg/mL showed no significant differences in their O.D. values (0.91, 0.79, and 0.53, respectively). For inhibition percentages, treatment with concentration 12.5 mg/mL gave the highest result (81.25%) followed by treatment at concentrations 6.25 and 9.38 mg/mL that showed no significant differences in their inhibition percentage (67.75% and 72.23%, respectively). Concentration 3.13 mg/mL resulted in 44.49% of biofilm inhibition and the positive control resulted in 30.75% of biofilm inhibition. Confocal laser scanning microscopy (CLSM) analysis of Xoo biofilm inhibition and breakdown showed the presence of non-viable Xoo cells and changes in aggregation size due to increase in AMMH leaf extract concentration. Control slides showed the absence of Xoo dead cells.

Resumo Xanthomonas oryzae pv. oryzae (Xoo), um patogênico responsável pela influência bacteriana na folha do arroz, produz biofilme para proteger células Xoo viáveis de agentes antimicrobianos. Foi conduzido um estudo para determinar a potência do extrato de folha de Acacia mangium methanol (AMMH) como um inibidor de biofilme Xoo. Quatro concentrações (3,13, 6,25, 9,38 e 12,5 mg/mL) de extrato de folha de AMMH foram testadas quanto à sua capacidade de inibir a formação de biofilme Xoo em uma placa de microtitulação de 96 poços. Os resultados mostraram que os controles negativos tiveram o maior valor de OD do que os outros tratamentos, indicando a intensa formação de biofilme. Isso foi seguido do controle positivo (sulfato de estreptomicina, com concentração de 0,2 mg/mL, e extrato de folha de AMMH, com concentração de 3,13 mg/mL), que não apresentou diferenças significativas nos seus valores OD (1,96 e 1,57, respectivamente). Todos os outros tratamentos com concentrações de 6,25, 9,38, e 12,5 mg/mL não tiveram diferenças significativas nos seus valores OD (0,91, 0,79, e 0,53, respectivamente). Para percentagens de inibição, o tratamento com concentração 12,5 mg/mL apresentou o maior resultado (81,25%), seguido do tratamento em concentrações de 6,25 e 9,38 mg/mL, que não mostraram diferenças significativas na sua percentagem de inibição (67,75 e 72,23%, respectivamente). Concentração 3,13 mg/mL resultou em 44,49% de inibição do biofilme, e o controle positivo resultou em 30,75% de inibição do biofilme. Análise por microscopia confocal de leitura a laser de inibição e separação de biofilme Xoo revelou a presença de células Xoo não viáveis e alterações no tamanho da agregação por causa do aumento na concentração de extrato de folha de AMMH. Slides de controle mostraram a ausência de células Xoo mortas.

Suppression of Xanthomonas oryzae pv. oryzae biofilm formation by Acacia mangium methanol leaf extract

Abstract Xanthomonas oryzae pv. oryzae (Xoo), a pathogen responsible for rice bacterial leaf blight, produces biofilm to protect viable Xoo cells from antimicrobial agents. A study was conducted to determine the potency of Acacia mangium methanol (AMMH) leaf extract as a Xoo biofilm inhibitor. Four concentrations (3.13, 6.25, 9.38, and 12.5 mg/mL) of AMMH leaf extract were tested for their ability to inhibit Xoo biofilm formation on a 96-well microtiter plate. The results showed that the negative controls had the highest O.D. values from other treatments, indicating the intense formation of biofilm. This was followed by the positive control (Streptomycin sulfate, 0.2 mg/mL) and AMMH leaf extract at concentration 3.13 mg/mL, which showed no significant differences in their O.D. values (1.96 and 1.57, respectively). All other treatments at concentrations of 6.25, 9.38, and 12.5 mg/mL showed no significant differences in their O.D. values (0.91, 0.79, and 0.53, respectively). For inhibition percentages, treatment with concentration 12.5 mg/mL gave the highest result (81.25%) followed by treatment at concentrations 6.25 and 9.38 mg/mL that showed no significant differences in their inhibition percentage (67.75% and 72.23%, respectively). Concentration 3.13 mg/mL resulted in 44.49% of biofilm inhibition and the positive control resulted in 30.75% of biofilm inhibition. Confocal laser scanning microscopy (CLSM) analysis of Xoo biofilm inhibition and breakdown showed the presence of non-viable Xoo cells and changes in aggregation size due to increase in AMMH leaf extract concentration. Control slides showed the absence of Xoo dead cells.

Resumo Xanthomonas oryzae pv. oryzae (Xoo), um patogênico responsável pela influência bacteriana na folha do arroz, produz biofilme para proteger células Xoo viáveis de agentes antimicrobianos. Foi conduzido um estudo para determinar a potência do extrato de folha de Acacia mangium methanol (AMMH) como um inibidor de biofilme Xoo. Quatro concentrações (3,13, 6,25, 9,38 e 12,5 mg/mL) de extrato de folha de AMMH foram testadas quanto à sua capacidade de inibir a formação de biofilme Xoo em uma placa de microtitulação de 96 poços. Os resultados mostraram que os controles negativos tiveram o maior valor de OD do que os outros tratamentos, indicando a intensa formação de biofilme. Isso foi seguido do controle positivo (sulfato de estreptomicina, com concentração de 0,2 mg/mL, e extrato de folha de AMMH, com concentração de 3,13 mg/mL), que não apresentou diferenças significativas nos seus valores OD (1,96 e 1,57, respectivamente). Todos os outros tratamentos com concentrações de 6,25, 9,38, e 12,5 mg/mL não tiveram diferenças significativas nos seus valores OD (0,91, 0,79, e 0,53, respectivamente). Para percentagens de inibição, o tratamento com concentração 12,5 mg/mL apresentou o maior resultado (81,25%), seguido do tratamento em concentrações de 6,25 e 9,38 mg/mL, que não mostraram diferenças significativas na sua percentagem de inibição (67,75 e 72,23%, respectivamente). Concentração 3,13 mg/mL resultou em 44,49% de inibição do biofilme, e o controle positivo resultou em 30,75% de inibição do biofilme. Análise por microscopia confocal de leitura a laser de inibição e separação de biofilme Xoo revelou a presença de células Xoo não viáveis e alterações no tamanho da agregação por causa do aumento na concentração de extrato de folha de AMMH. Slides de controle mostraram a ausência de células Xoo mortas.

Identification of phenolic compounds and evaluation of antibacterial properties of Piper sarmentosum Roxb. against rice pathogenic bacteria

Aims: Piper sarmentosum Roxb. has a long history of medicinal usage and potential in treating many diseases and ailments. It is known for its medicinal properties and contains a variety of active chemical compounds. This study was conducted to identify and quantify the phenolic compounds of the leaf and fruit extract of P. sarmentosum, as well as their antibacterial activities. Methodology and results: High Performance Liquid Chromatography (HPLC) was carried out to identify the phenolic compounds in the samples. Antibacterial performance of the samples was measured using agar well and disc diffusion assay, where its MIC values were then determined. After going through HPLC-UV, the major phenolic compounds identified in both extracts were tannic acid, gallic acid, quercetin and naringin. The leaf and fruit of P. sarmentosum exhibited moderate to strong antibacterial activity when tested against pythopathogenic bacteria with an inhibition range of 10.67-17.33 mm at 100 mg/mL. Conclusion, significance and impact of study: The leaf and fruit extracts of P. sarmentosum were proven to have effective inhibitory effects on Pseudomonas fuscovaginae and Xanthomonas oryzae pv. oryzae, which are also the causal agents of sheath brown rot and bacterial leaf blight in rice. This is believed to be due to the presence of the phenolic compounds in these samples. In a subsequent study, the researchers are planning to apply a formulation, developed from the crude extract, in the glasshouse and field trial.

Antimicrobial activity and identification of potential antimicrobial compounds from aquatic pteridophyte, Azolla microphylla Kaulf.

Azolla microphylla Kaulf. is an aquatic nitrogen fixing pteridophyte commonly found in aquatic habitats including paddy fields. Methanolic extract of the fronds of A. microphylla was subjected to partial purification by solvent partitioning with diethyl ether and ethyl acetate followed by hydrolysis, and further partitioning with ethyl acetate. The two fractions, thus obtained were tested for antibacterial activity. It was observed that the ethyl acetate fraction inhibited the growth of the pathogenic bacterium Xanthomonas oryzae. The GC-MS analysis of the ethyl acetate fraction showed several prominent peaks with retention time ranging from 8.83 to 45.54 min. A comparison of these peaks with the GC-MS libraries revealed that it could be eicosenes and heptadecanes with potential of antimicrobial activity.

Hpa1 harpin needs nitroxyl terminus to promote vegetative growth and leaf photosynthesis in Arabidopsis.

Hpa1 is a harpin protein produced by Xanthomonas oryzae, an important bacterial pathogen of rice, and has the growth-promoting activity in plants. To understand the molecular basis for the function of Hpa1, we generated an inactive variant protein, Hpa1ΔNT, by deleting the nitroxyl-terminal region of the Hpa1 sequence and compared Hpa1ΔNT with the full-length protein in terms of the effects on vegetative growth and related physiological responses in Arabidopsis. When Hpa1 was applied to plants, it acted to enhance the vegetative growth but did not affect the floral development. Enhanced plant growth was accompanied by induced expression of growth-promoting genes in plant leaves. The growth-promoting activity of Hpa1 was further correlated with a physiological consequence shown as promoted leaf photosynthesis as a result of facilitated CO2 conduction through leaf stomata and mesophyll cells. On the contrary, plant growth, growth-promoting gene expression, and the physiological consequence changed little in response to the Hpa1ΔNT treatment. These analyses suggest that Hpa1 requires the nitroxyl-terminus to facilitate CO2 transport inside leaf cells and promote leaf photosynthesis and vegetative growth of the plant.

Down-regulation of OsSAG12-1 results in enhanced senescence and pathogen-induced cell death in transgenic rice plants.

Senescence is a highly regulated process accompanied by changes in gene expression. While the mRNA levels of most genes decline, the mRNA levels of specific genes (senescence associated genes, SAGs) increase during senescence. Arabidopsis SAG12 (AtSAG12) gene codes for papain-like cysteine protease. The promoter of AtSAG12 is SA-responsive and reported to be useful to delay senescence by expressing cytokinin biosynthesis gene isopentenyltransferase specifically during senescence in several plants including Arabidopsis, lettuce and rice. The physiological role of AtSAG12 is not known; the homozygous atsag12 mutant neither fails to develop senescenceassociated vacuoles nor shows any morphological phenotype. Through BLAST search using AtSAG12 amino acid sequences as query, we identified a few putative homologues from rice genome (OsSAGs; Oryza sativa SAGs). OsSAG12-1 is the closest homologue of AtSAG12 with 64% similar amino acid composition. Expression of OsSAG12-1 is induced during senescence and pathogen-induced cell death. To evaluate the possible role of OsSAG12-1 we generated RNAi transgenic lines in Japonica rice cultivar TP309. The transgenic lines developed early senescence at varying levels and showed enhanced cell death when inoculated with bacterial pathogen Xanthomonas oryzae pv.oryzae. Our results suggest that OsSAG12-1 is a negative regulator of cell death in rice.

Genetic Diversity of Xanthomonas oryzae pv.oryzae Strains from China,Japan and Philippines / 微生物学通报

The IS-PCR and Rep-PCR were used to analyze 19 strains of Xanthomonas oryzae pv.oryzae from China,Japan and Philippines.Four specific primers,especially,IS1113 and ERIC could distinguish the strains from different countries.Using UPGMA analysis,most strains were in group 2 and 3.No relationship were observed between UPGMA groups and pathotypes.