Cadmium stress alters the redox reaction and hormone balance in oilseed rape (Brassica napus L.) leaves

In order to understand the physiological response of oilseed rape (Brassica napus L.) leaves to cadmium (Cd) stress and exploit the physiological mechanisms involved in Cd tolerance, macro-mineral and chlorophyll concentrations, reactive oxygen species (ROS) accumulation, activities of enzymatic antioxidants, nonenzymatic compounds metabolism, endogenous hormonal changes, and balance in leaves of oilseed rape exposed to 0, 100, or 200 μM CdSO4 were investigated. The results showed that under Cd exposure, Cd concentrations in the leaves continually increased while macro-minerals and chlorophyll concentrations decreased significantly. Meanwhile, with increased Cd stress, superoxide anion (O 2 • − ) production rate and hydrogen peroxide (H2O2) concentrations in the leaves increased significantly, which caused malondialdehyde (MDA) accumulation and oxidative stress. For scavenging excess accumulated ROS and alleviating oxidative injury in the leaves, the activity of enzymatic antioxidants, such as superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), was increased significantly at certain stress levels. However, with increased Cd stress, the antioxidant enzyme activities all showed a trend towards reduction. The nonenzymatic antioxidative compounds, such as proline and total soluble sugars, accumulated continuously with increased Cd stress to play a long-term role in scavenging ROS. In addition, ABA levels also increased continuously with Cd stress while ZR decreased and the ABA/ZR ratio increased, which might also be providing a protective role against Cd toxicity.

Genome-wide association analysis and differential expression analysis of resistance to Sclerotinia stem rot in Brassica napus

Brassica napus is one of the most important oil crops in the world, and stem rot caused by the fungus Sclerotinia sclerotiorum results in major losses in yield and quality. To elucidate resistance genes and pathogenesis-related genes, genome-wide association analysis of 347 accessions was performed using the Illumina 60K Brassica SNP (single nucleotide polymorphism) array. In addition, the detached stem inoculation assay was used to select five highly resistant (R) and susceptible (S) B. napus lines, 48 h postinoculation with S. sclerotiorum for transcriptome sequencing. We identified 17 significant associations for stem resistance on chromosomes A8 and C6, five of which were on A8 and 12 on C6. The SNPs identified on A8 were located in a 409-kb haplotype block, and those on C6 were consistent with previous QTL mapping efforts. Transcriptome analysis suggested that S. sclerotiorum infection activates the immune system, sulphur metabolism, especially glutathione (GSH) and glucosinolates in both R and S genotypes. Genes found to be specific to the R genotype related to the jasmonic acid pathway, lignin biosynthesis, defence response, signal transduction and encoding transcription factors. Twenty-four genes were identified in both the SNP-trait association and transcriptome sequencing analyses, including a tau class glutathione S-transferase (GSTU) gene cluster. This study provides useful insight into the molecular mechanisms underlying the plant's response to S. sclerotiorum.

The role of spiders as predators of two lepidopteran Brassica pests

Spiders are thought to play a significant role in limiting pest outbreaks in agroecosystems such as vineyards, orchards and cotton. The diversity and impact of spiders in vegetable crops are less well understood, although there is evidence that predators may be important for suppression of lepidopteran pests in Brassica crops, particularly early in the season before parasitoids become established. Sampling was conducted in early season plantings of Brassicas in the Lockyer Valley (South East Queensland, Australia) in order to determine the most commonly occurring spider families. The most numerous were Theridiidae, which were more strongly associated with cauliflower and poorly associated with cabbage. The Lycosidae and Clubionidae/Miturgidae (formerly in the ‘catch-all’ family Clubionidae) also occurred commonly. Lycosidae (and to a lesser extent Salticidae) had above average abundance in Chinese cabbage and below average abundance in broccoli compared with average abundance for these spider families; Clubionidae/Miturgidae had above average abundance in cauliflower. Laboratory studies were then conducted to explore the predatory capacity of these three most commonly occurring spider families. All three were capable of feeding on larvae of the diamondback moth, Plutella xylostella (Linnaeus), and cabbage cluster caterpillar, Crocidolomia pavonana (Fabricius), under laboratory conditions. Theridiidae, which are thought to prey on small pests such as leafhoppers and aphids, were able to successfully attack larvae up to five times their body size. Predation rates varied from an average of 1.7 (SE = 0.47) (1.6 control corrected) larvae consumed over a 24 h period in the case of the Theridiidae, to 3.3 (SE = 0.60) larvae for the Clubionidae/Miturgidae.

Mode of Action of a Purified Antitumor Protein from the Proteinaceous Crystal of Bacillus thuringiensis subsp. thuringiensis on Yoshida Ascites Sarcoma Cells

A purified antitumor protein from the proteinaceous crystal of Bacillus thuringiensis subsp. thuringiensis inhibits the growth of Yoshida ascites sarcoma both in vivo and in vitro. Exogenous respiration of the tumor cells was unaffected by the protein at a concentration as high as 500 µg/ml. The antitumor protein inhibits the uptake and incorporation of labeled precursors into macromolecules. However, the ratio of incorporation over uptake is not affected by the protein. Further, the protein brings about the leakage of 260-nm-absorbing material, proteins, and 32P-labeled cellular constituents from the Yoshida ascites sarcoma cells. The results show that the action of the antitumor protein appears to alter the cellular permeability of the tumor cells.

Quorum sensing in the plant pathogen Erwinia carotovora subsp. carotovora

Erwinia carotovora subsp. carotovora (Ecc) is a Gram-negative enterobacterium that causes soft-rot in potato and other crops. The main virulence determinants, the extracellular plant cell wall -degrading enzymes (PCWDEs), lead to plant tissue maceration. In order to establish a successful infection the production of PCWDEs are controlled by a complex regulatory network, including both specific and global activators and repressors. One of the most important virulence regulation systems in Ecc is mediated by quorum sensing (QS), which is a population density -dependent cell-to-cell communication mechanism used by many Gram-negative bacteria. In these bacteria N-acylhomoserine lactones (AHSL), act as diffusible signaling molecules enabling communication between bacterial cells. The AHSLs are structurally diverse and differ in their acyl chain length. This gives the bacteria signaling specificity and enables the recognition and communication within its own species. In order to detect and respond to the AHSLs the bacteria use QS regulators, LuxR-type proteins. The aim of this study was to get a deeper understanding of the Ecc QS system. In the first part of the study we showed that even different strains of Ecc use different dialects and of physiological concentrations, only the cognate AHSL with the correct acyl chain is recognized as a signal that can switch on virulence genes. The molecular basis of the substrate specificity of the AHSL synthase ExpI was investigated in order to recognize the acyl chain length specificity determinants of distinct AHSL synthases. Several critical residues that define the size of the substrate-binding pocket were identified. We demonstrated that in the ExpISCC1 mutations M127T and F69L are sufficient to change the N-3-oxohexanoyl-L-homoserine lactone producing ExpISCC1 to an N-3-oxooctanoyl-L-homoserine lactone (3-oxo-C8-HSL) producing enzyme. In the second study the means of sensing specificity and response to the AHSL signaling molecule were investigated. We demonstrated that the AHSL receptor ExpR1 of Ecc strain SCC3193 has strict specificity for the cognate AHSL 3-oxo-C8-HSL. In addition we identified a second AHSL receptor ExpR2 with a novel property to sense AHSLs with different acyl chain lengths. In the absence of AHSLs ExpR1 and ExpR2 were found to act synergistically to repress the virulence gene expression. This repression was shown to be released by addition of AHSLs and appears to be largely mediated by the global negative regulator RsmA. In the third study random transposon mutagenesis was used to widen the knowledge of the Ecc QS regulon. Two new QS-controlled target genes, encoding a DNA-binding regulator Hor and a plant ferredoxin-like protein FerE, were identified. The QS control of the identified genes was executed by the QS regulators ExpR1 and ExpR2 and as expression of PCWDE genes mediated by the RsmA repressor. Hor was shown to contribute to bacterial virulence at least partly through its control of PCWDE production, while FerE was shown to contribute to oxidative stress tolerance and in planta fitness of the bacteria. In addition our results suggest that QS is central to the control of oxidative stress tolerance in Ecc. In conclusion, these results indicate that Ecc strain SCC3193 is able to react and respond both to the cognate AHSL signal and the signals produced by other bacterial species, in order to control a wide variety of functions in the plant pathogen Ecc.

Disease burden due to Streptococcus dysgalactiae subsp equisimilis (group G and C streptococcus) is higher than that due to Streptococcus pyogenes among Mumbai school children

Streptococcus pyogenes [group A streptococcus (GAS)], a human pathogen, and Streptococcus dysgalactiae subsp. equisimilis [human group G and C streptococcus (GGS/GCS)] are evolutionarily related, share the same tissue niche in humans, exchange genetic material, share up to half of their virulence-associated genes and cause a similar spectrum of diseases. Yet, GGS/GCS is often considered as a commensal bacterium and its role in streptococcal disease burden is under-recognized. While reports of the recovery of GGS/GCS from normally sterile sites are increasing, studies describing GGS/GCS throat colonization rates relative to GAS in the same population are very few. This study was carried out in India where the burden of streptococcal diseases, including rheumatic fever and rheumatic heart disease, is high. As part of a surveillance study, throat swabs were taken from 1504 children attending 7 municipal schools in Mumbai, India, during 2006-2008. GAS and GGS/GCS were identified on the basis of beta-haemolytic activity, carbohydrate group and PYR test, and were subsequently typed. The GGS/GCS carriage rate (1166/1504, 11%) was eightfold higher than the GAS carriage (22/1504, 1.5%) rate in this population. The 166 GGS/GCS isolates collected represented 21 different emm types (molecular types), and the 22 GAS isolates represented 15 different emm types. Although the rate of pharyngitis associated with GGS/GCS is marginally lower than with GAS, high rates of throat colonization by GGS/GCS underscore its importance in the pathogenesis of pharyngitis.

Genetic transformation of Brassica nigra by agrobacterium based vector and direct plasmid uptake

Genetic transformation systems have been established for Brassica nigra (cv. IC 257) by using an Agrobacterium binary vector as well as by direct DNA uptake of a plasmid vector. Both the type of vectors carried nptII gene and gus gene. For Agrobacterium mediated transformation, hypocotyl tissue explants were used, and up to 33% of the explants produced calli on selection medium. All of these expressed B-glucuronidase gene on histochemical staining. Protoplasts isolated from hypocotyl tissues of seedlings could be transformed with a plasmid vector by FEG mediated uptake of vector DNA. A number of fertile kanamycin resistant plants were obtained using both the methods, and their transformed nature was confirmed by Southern blot analysis and histochemical staining for GUS. Backcrossed and selfed progenies of these transformed plants showed the presence of npt and gus genes.

TolA mediates the differential detergent resistance pattern between the Salmonella enterica subsp enterica serovars Typhi and Typhimurium

The tol-pal genes are essential for maintaining the outer membrane integrity and detergent resistance in various Gram-negative bacteria, including Salmonella. The role of TolA has been well established for the bile resistance of Salmonella enterica subsp. enterica serovar Typhimurium. We compared the bile resistance pattern between the S. enterica serovars Typhi and Typhimurium and observed that Typhi is more resistant to bile-mediated damage. A closer look revealed a significant difference in the TolA sequence between the two serovars which contributes to the differential detergent resistance. The tolA knockout of both the serovars behaves completely differently in terms of membrane organization and morphology. The role of the Pal proteins and difference in LPS organization between the two serovars were verified and were found to have no direct connection with the altered bile resistance. In normal Luria broth (LB), S. Typhi Delta tolA is filamentous while S. Typhimurium Delta tolA grows as single cells, similar to the wildtype. In low osmolarity LB, however, S. Typhimurium Delta tolA started chaining and S. Typhi Delta tolA showed no growth. Further investigation revealed that the chaining phenomenon observed was the result of failure of the outer membrane to separate in the dividing cells. Taken together, the results substantiate the evolution of a shorter TolA in S. Typhi to counteract high bile concentrations, at the cost of lower osmotic tolerance.

Efecto de policultivos repollo-arroz, repollo-zanahoria, repollo-frijol, repollo-cebolla sobre la entomofauna en el cultivo del repollo (Brassica oleraceae L.) híbrido Izalco

El experimento de campo se llevó a cabo en San Ignacio, municipio de la Concepción, departamento de Masaya con el objetivo de evaluar el efecto de policultivos (repollo-arroz, repollo-zanahoria, repollo-frijol y repollo-cebolla) sobre la palomilla de la col (Plutella xylostella L.) en el cultivo de repollo (Brassica oleracea L.) híbrido Izalco, además se evaluó un tratamiento en monocultivo de repollo y otro con aplicación de insecticida biológico dipel (Bacillus thuringiensis) para ser usados como testigo absoluto y relativo respectivamente. Las plagas predominantes durante el ensayo fueron Plutella Xylostella, Spodoptera sp., Ascia monuste y Afidos. En la etapa de crecimiento vegetativo (0-32 días después del trasplante, DDT),las población de Plutella xylostella fue baja, pero en la etapa de preformación de cabeza (32-64 DDT) la población se incrementó, los tratamientos que presentaron menores poblaciones fueron repollo-arroz, repollo + dipel seguido de repollo-cebolla. En la etapa de formación de cabeza (64-88 DDT) la incidencia de la plaga fue siempre alta en el monocultivo de repollo y repollo-frijol a excepción de los demás tratamientos, siendo esta la etapa de mayor importancia del cultivo. El uso de cultivos en asocio no ejerció ningún efecto sobre Ascia monuste, Spodoptera sp. y Afidos. En la incidencia de insectos benéficos sólo se encontró & ralas y sus poblaciones se mantuvieron a niveles bajos.

Evaluación del rendimiento agronómico de doce cultivares de repollo (Brassica oleracea L) y la incidencia de Plutella xylostella L, en la Estación experimental Raul González, valle de Sébaco, Matagalpa

El estudio se realizó en la Estación experimental Raúl González del valle de Sébaco de Junio a Septiembre de 1994. Con el objetivo de determinar las características agronómicas de cada cultivar de repollo (Brasica olearacea L), para resolver algunos problemas de pequeños y medianos productores que demandan cultivares con buen rendimiento y resistente a plagas y enfermedades. Se evaluaron las variedades: Gluckstadter Mittelfrüher, Yeshen, Migthy YR, Copenhagen Market, Conquest, Izalco, Fortuna, Grenadier, Discovery, Giant, Superette YR y Glory of Enkhuizen. El diseño utilizado fue de Bloques Completos al Azar (B.C.A) con cuatro repeticiones, evaluándose las variables de crecimiento y desarrollo del cultivo, así como lo relacionado al rendimiento agronómico y la incidencia de Plutella xylostella L. Los datos obtenidos se sometieron al análisis de varianza y a la prueba de Tukey. Las variedades de mejor crecimiento y desarrollo fueron: Yeshen, Migthy YR, Superette YR, Fortuna e Izalco. En las variables de calidad no se obtuvo diferencia significativa., sin embargo, los cultivares Superette YR, Izalco y Fortuna resultaron con el mayor índice de compactación. Respecto al rendimiento los cultivares Grenadier, Izalco y Fortuna obtuvieron el mayor porcentaje de formación de cabezas así como, el mayor peso de cabeza por hectárea. Los insecticidas utilizados para el manejo de P xylostella no lograron reducir sus poblaciones durante las etapas de preformación y llenado de cabezas. Al finalizar el ciclo del cultivo se presentó Xanthomonas campestris p. y campestris resultando tolerantes los cultivares Grenadier, Discovery, Izalco, Migthy YR y Fortuna; perdiendo casi la totalidad de su población las variedades: Glory of Enkhuizen, Copenhagen Market y Conquest.

Evaluación del rendimiento agronómico de seis cultivares de repollo (Brassica oleracea L.) en la Estación Experimental "San José de las Latas", Jinotega

En el presente trabajo se evaluó el rendimiento agronómico de seis cultivares Brassica oleracea L. var. caPitata; Summer Autumn, Mighty YR, Perfect Ball, Mighty, Yessen #-631-4, Yessen # 33-18. El ensayo se estableció en la estacion las " Latas " Ubicada en Jinotega a 1, 400 m.s.n.m. con una precipitación media anual de 2.291 mm, y una temperatura promedio 21.86 ºC. La siembra se realizó en época de primera del año 1990. Empleándose un diseño Bloques al Azar (BCA) Con De los resultados obtenidos, se encontró cultivares que presentaron mayor crecimiento y desarrollo estuvo en primer lugar el cultivar Summer Auttumn, en segundo la linea Yessen # 631-4, y en tercer lugar la linea Yessen # 33-18. Respecto a la calidad del producto comercial, sus variables se mantuvieron estadísticamente sin diferencias significativas a pesar de diferir en cultivares que no presentaron buen desarrollo. El cultivar Summer, menor consistencia, seguida de la línea Yessen # 33-18. Al evaluar el rendimiento, se encontró que el cultivar Summer Autumn y la linea Yessen # 631-4 obtuvieron los mejores promedios. Seguidos por el cultivar Mighty YR y la línea de Yessen # 33-18, aunque Mighty YR presento baja tolerancia a la bacteriosis (Xanthomonas campestris P.).

Evaluación de criterios de aplicación para el manejo de Plutella xylostella (L) en repollo (Brassica oleracea L.)

Con el objetivo de evaluar diferentes criterios de aplicación para el manejo de Plutella xylostella (L.) en el cultivo de repollo se realizó un experimento de campo en la época de primera en el Centro experimental de campos Azules (Masatepe, Departamento de Masaya), se mantuvieron diferentes niveles de esta plaga por medio de aplicaciones semanales del insecticida Chlorfluazurón (Júpiter) a una dosis de 182 cc/ha. La incidencia de la plaga no fue uniforme durante el ciclo del cultivo, oscilando por encima de los niveles establecidos a partir de los 28 DDT. La mayor incidencia fue de 1.2 larvas/planta en la etapa de formación de cabeza y de 3 3 larvas/planta para la etapa de llenado de cabeza afectando el área foliar, precio por cabeza, y los ingresos económicos. A los resultados agronómicos se les aplicó un análisis económico, resultando del aná11s1s de retorno marginal, tasas de retornos marginales

Evaluación de productos botánicos y biológicos para el control de plagas defoliadoras en el cultivo del repollo (Brassica oleracea) híbrido Izalco

Durante el ciclo del cultivo se presentaron plagas como: P. xylostella, Spodoptera sp. y Helulla phydeliales. Presentando mayor incidencia P. Jlylastello con un comportamiento diferente en cada etapa del cultivo en contrando que en las etapas de formación y llenado de cabezas (30-50 y 50-80 DDT) se presentaron las mayores poblaciones, considerando estas dos etapas corno períodos críticos, ya que la plaga en esta etapa afecta la calidad del producto lo que influye económicamente. Durante el ciclo del cultivo fue notorio el control eficaz que ejercio el tratamiento Extracto Acuoso de Semilla de Neem sobre P. xylostella; seguido por los tratamientos javelin (B. thuringiensis) y júpiter (Chlorfluasuron) que lograron mantener las poblaciones bajas, no así los tratamientos Dipel comercial, BT. Nacional y Extracto Acuoso de Semilla de Mamey los que mantuvieron poblaciones similares a las presentadas por el testigo (sin aplicación). El análisis económico de los tratamientos, indicó que el tratamiento de mayor rentabilidad fue el Extracto Acuoso de Semilla de Neem seguido por el tratamiento júpiter, los cuales poseen una tasa de retorno marginal aceptable en cambio los tratamientos, Dipel Comercial, Extracto Acuoso de Semilla de Mamey, BT Nacional no son recomendables para el control de P. xylostella.

Evaluación del protectante nufilm-17 en la efectividad de los insecticidas botánicos y biológicos sobre la entomofauna en el cultivo del repollo (Brassica oleraceae L.) hibrido izalco en época de apante

Con el objetivo de evaluar el efecto del protectante Nufilm-17 sobre la efectividad de los productos Botánicos y Biológicos contra Plutella xylostella L. en el cultivo de repollo se llevó a cabo el presente trabajo en el Centro Experimental del valle de Sébaco "Raúl Gonzalez”, de Dic.91 - Marzo 92. El diseño utilizado fué de Bloques Completos al Azar (BCA), con 4 repeticiones y 7 tratamientos que fueron distribuidos al azar en el campo: las variables a medir fueron: número de larvas de P. xylostella números de arañas, números de polybias porcentaje de parasitismo de Diadegma insularis sobre larvas de P. xylostella números de cabezas/ha. precio/cabeza, daño foliar. Además se hizo análisis económico para conocer la rentabilidad del uso de las alternativas en el control de las principales plaga en el cultivo. Los recuento se hicieron cada 8 días hasta el final de la cosecha. El comportamiento de P. xylostella fue diferente en la distintas etapas del cultivo siendo la etapa de crecimiento vegetativo y formación de cabeza (0-30 y 30-60 DDT) donde se presentaron las mayores poblaciones. Siendo notorio el control que efectuo el tratamiento Nim + Nufilm-17 sobre P. xylostella que logró mantener las poblaciones bajas. El porcentaje de parasitismo observado en larvas de P. xylostella registró valores entre 5 y 30 %. El análisis económico, demostró que los tratamientos Nim + Nufilm-17 y B. thuringiesis presentaron mayor rentabilidad teniendo una tasa de retorno marginal superior a la tasa comparativa que es de 125 %.

Evaluación de productos botánicos y biológicos para el control de Plutella xylostella en el cultivo del repollo (Brassica oleraceae L.) hibrido Izalco en época de apante

Con el fin de encontrar el mejor producto para el control de Plutella xylostella en el cultivo del repollo, se llevó a cabo un ensayo a nivel de campo en época seca (Die 91-Mar 92) en el valle de Sébaco-Matagalpa donde se evaluaron los tratamientos Bacillus thuringiensis nacional, Dipel comercial, Javelin, Nim, Mamey y Júpiter, con un criterio de aplicación de 0.5 larvas por plantas. Las poblaciones de Plutella xylostella fueron relativamente bajas en la etapa de crecimiento vegetativo disminuyendo en las últimas dos etapas formación y llenado de cabeza. Durante las tres fases del cultivo las poblaciones de Plutella xylostella fueron menores en los tratamientos Javelin Mamey y Júpiter. Los insecticidas Dipel. Javelin y Mamey fueron diferentes estadísticamente del testigo (sin aplicación), según el análisis económico los primeros dos insecticidas son rentables económicamente sucediendo lo contrario con el insecticida Mamey por lo tanto su uso no es recomendado para el manejo de la plaga. El insecticida Júpiter puede ser utilizado contra la plaga ya que muestra una tasa de retorno marginal superior a la tasa comparativa. La presencia de enemigos naturales (Araña, Polybia sppy el parasitoide Diadegma insularis) fue notoria en el cultivo, cabe señalar su importancia ya que intervienen en el equilibrio del ecosistema dentro del cultivo (plaga-repollo). Estos predadores no se vieron afectados por las aplicaciones de los insecticidas botánicos y biológicos evaluados en el cultivo. El uso de estos productos parecen ser una alternativa de manejo de las plagas del cultivo a largo plazo ya que el establecimiento de la fauna benéfica en el sistema y el no uso de productos sintéticos evitan los problemas de resistencia en las plagas y la contaminación ambiental.