Impact of nitrogen and phosphorus fertilizers on Cry1Ac protein contents in transgenic cotton / Impacto de fertilizantes de nitrogênio e fósforo nos conteúdos de proteína Cry1Ac em algodão transgênico

Application of different fertilizers to check the efficiency of expression of Bt (Bacillus thuringiensis) gene in one of the leading commercialized crops (cotton) against Lepidopteran species is of great concern. The expression of Cry protein level can be controlled by the improvement of nutrients levels. Therefore, the myth of response of Cry toxin to different combinations of NP fertilizers was explored in three Bt cotton cultivars. Combinations include three levels of nitrogen and three levels of phosphorus fertilizers. Immunostrips and Cry gene(s) specific primer based PCR (Polymerase Chain Reaction) analysis were used for the presence of Bt gene that unveiled the presence of Cry1Ac gene only. Further, the ELISA (enzyme-linked immunosorbent assay) kit was used to quantify the expression of Cry1Ac protein. Under various NP fertilizers rates, the level of toxin protein exhibited highly significant differences. The highest toxin level mean was found to be 2.3740 and 2.1732 µg/g under the treatment of N150P75 kg ha-¹ combination while the lowest toxin level mean was found to be 0.9158 and 0.7641 µg/g at the N50P25 kg ha-¹ level at 80 and 120 DAS (Days After Sowing), respectively. It was concluded from the research that the usage of NP fertilizers has a positive relation with the expression of Cry1Ac toxin in Bt cotton. We recommend using the N150P50 kg ha-1 level as the most economical and practicable fertilizer instead of the standard dose N100P50 kg ha-¹ to get the desired level of Cry1Ac level for long lasting plant resistance (<1.5). The revised dose of fertilizer may help farmers to avoid the cross-resistance development in contradiction of insect pests.

A aplicação de diferentes fertilizantes para verificar a eficiência da expressão do gene Bt (Bacillus thuringiensis) em uma das principais culturas comercializadas (algodão) contra espécies de lepidópteros é uma grande preocupação. A expressão do nível de proteína Cry pode ser controlada pela melhoria dos níveis de nutrientes. Portanto, o mito da resposta da toxina Cry a diferentes combinações de fertilizantes NP foi explorado em três cultivares de algodão Bt. As combinações incluem três níveis de nitrogênio e três níveis de fertilizantes de fósforo. A análise de PCR (reação em cadeia da polimerase) específica para o gene (s) Immunostrips e Cry (s) foi usada para a presença do gene Bt que revelou a presença do gene Cry1Ac apenas. Além disso, o kit ELISA (ensaio de imunoabsorção enzimática) foi usado para quantificar a expressão da proteína Cry1Ac. Sob várias taxas de fertilizantes NP, o nível de proteína de toxina exibiu diferenças altamente significativas. A média do nível mais alto de toxina foi de 2,3740 e 2,1732 µg / g sob o tratamento da combinação N150P75 kg ha-¹, enquanto a média do nível mais baixo de toxina foi de 0,9158 e 0,7641 µg / g no nível de N50P25 kg ha-¹ em 80 e 120 DAS (dias após a semeadura), respectivamente. Concluiu-se com a pesquisa que o uso de fertilizantes NP tem relação positiva com a expressão da toxina Cry1Ac no algodão Bt. Recomendamos o uso do nível de N150P50 kg ha-¹ como o fertilizante mais econômico e praticável em vez da dose padrão N100P50 kg ha-¹ para obter o nível desejado de nível de Cry1Ac para resistência de planta de longa duração (<1,5). A dose revisada de fertilizante pode ajudar os agricultores a evitar o desenvolvimento de resistência cruzada em contradição com as pragas de insetos.

Impact of nitrogen and phosphorus fertilizers on Cry1Ac protein contents in transgenic cotton / Impacto de fertilizantes de nitrogênio e fósforo nos conteúdos de proteína Cry1Ac em algodão transgênico

Abstract Application of different fertilizers to check the efficiency of expression of Bt (Bacillus thuringiensis) gene in one of the leading commercialized crops (cotton) against Lepidopteran species is of great concern. The expression of Cry protein level can be controlled by the improvement of nutrients levels. Therefore, the myth of response of Cry toxin to different combinations of NP fertilizers was explored in three Bt cotton cultivars. Combinations include three levels of nitrogen and three levels of phosphorus fertilizers. Immunostrips and Cry gene(s) specific primer based PCR (Polymerase Chain Reaction) analysis were used for the presence of Bt gene that unveiled the presence of Cry1Ac gene only. Further, the ELISA (enzyme-linked immunosorbent assay) kit was used to quantify the expression of Cry1Ac protein. Under various NP fertilizers rates, the level of toxin protein exhibited highly significant differences. The highest toxin level mean was found to be 2.3740 and 2.1732 µg/g under the treatment of N150P75 kg ha-1 combination while the lowest toxin level mean was found to be 0.9158 and 0.7641 µg/g at the N50P25 kg ha-1 level at 80 and 120 DAS (Days After Sowing), respectively. It was concluded from the research that the usage of NP fertilizers has a positive relation with the expression of Cry1Ac toxin in Bt cotton. We recommend using the N150P50 kg ha-1 level as the most economical and practicable fertilizer instead of the standard dose N100P50 kg ha-1 to get the desired level of Cry1Ac level for long lasting plant resistance (<1.5). The revised dose of fertilizer may help farmers to avoid the cross-resistance development in contradiction of insect pests.

Resumo A aplicação de diferentes fertilizantes para verificar a eficiência da expressão do gene Bt (Bacillus thuringiensis) em uma das principais culturas comercializadas (algodão) contra espécies de lepidópteros é uma grande preocupação. A expressão do nível de proteína Cry pode ser controlada pela melhoria dos níveis de nutrientes. Portanto, o mito da resposta da toxina Cry a diferentes combinações de fertilizantes NP foi explorado em três cultivares de algodão Bt. As combinações incluem três níveis de nitrogênio e três níveis de fertilizantes de fósforo. A análise de PCR (reação em cadeia da polimerase) específica para o gene (s) Immunostrips e Cry (s) foi usada para a presença do gene Bt que revelou a presença do gene Cry1Ac apenas. Além disso, o kit ELISA (ensaio de imunoabsorção enzimática) foi usado para quantificar a expressão da proteína Cry1Ac. Sob várias taxas de fertilizantes NP, o nível de proteína de toxina exibiu diferenças altamente significativas. A média do nível mais alto de toxina foi de 2,3740 e 2,1732 µg / g sob o tratamento da combinação N150P75 kg ha-1, enquanto a média do nível mais baixo de toxina foi de 0,9158 e 0,7641 µg / g no nível de N50P25 kg ha-1 em 80 e 120 DAS (dias após a semeadura), respectivamente. Concluiu-se com a pesquisa que o uso de fertilizantes NP tem relação positiva com a expressão da toxina Cry1Ac no algodão Bt. Recomendamos o uso do nível de N150P50 kg ha-1 como o fertilizante mais econômico e praticável em vez da dose padrão N100P50 kg ha-1 para obter o nível desejado de nível de Cry1Ac para resistência de planta de longa duração (<1,5). A dose revisada de fertilizante pode ajudar os agricultores a evitar o desenvolvimento de resistência cruzada em contradição com as pragas de insetos.

Physicochemical features, functional characteristics, and health benefits of cottonseed oil: a review / Benefícios à saúde humana e características físico-químicas e funcionais do óleo de semente de algodão: uma revisão

Vegetable oils have their specific physicochemical properties due to which they are playing vital role in human nutritional diet for health benefits. Cottonseed oil is obtained from various species of cotton seeds that are famous to be grown mainly for their fiber quality. The most prominently used specie is Gossypium hirsutum. It is obvious that the seeds of different variety of cotton vary as grown in diverse agroclimatic conditions with respect to oil, fats and protein contents. Cottonseed oil is routinely used for cooking and food manufacturing products. Cottonseed oil obtained after proper extraction/processing steps from crude state to refined oil in a variety of ways. Cotton crop is considered for their dual-use purpose, for fiber quality and oil production to promote health benefits in the world. Keeping in view the above facts, this review clearly demonstrated an overview about physicochemical and functional properties of cottonseed oil to promote health benefits associated with the use of this oil. The overall characteristics and all concerned health benefits of CSO will further improve their usefulness is a compact way. We have summarized a brief multi-dimensional features of CSO in all aspects up to the best of our knowledge fort he end researchers who can further research in the respective aspect.

Os óleos vegetais têm propriedades físico-químicas específicas que desempenham um papel vital na dieta nutricional humana em benefício à saúde. O óleo de semente de algodão, utilizado rotineiramente no preparo e na fabricação de alimentos, é obtido através de várias espécies de sementes de algodão, famosas pela alta qualidade de sua fibra, cuja espécie mais utilizada é Gossypium hirsutum. As sementes variam, em relação ao teor do óleo, da gordura e das proteínas, de acordo com o cultivo e as diversas condições agroclimáticas. O óleo de algodão é obtido após etapas adequadas de extração das sementes e processamento do estado bruto ao refinado. Assim, a presente revisão demonstra, de maneira geral, as propriedades físico-químicas e funcionais do óleo de semente de algodão e seus benefícios à saúde humana, resumindo suas características multidimensionais. As características gerais e todos os benefícios do composto podem melhorar ainda mais se utilizadas de forma compacta, auxiliando futuros pesquisadores.

Physicochemical features, functional characteristics, and health benefits of cottonseed oil: a review / Benefícios à saúde humana e características físico-químicas e funcionais do óleo de semente de algodão: uma revisão

Vegetable oils have their specific physicochemical properties due to which they are playing vital role in human nutritional diet for health benefits. Cottonseed oil is obtained from various species of cotton seeds that are famous to be grown mainly for their fiber quality. The most prominently used specie is Gossypium hirsutum. It is obvious that the seeds of different variety of cotton vary as grown in diverse agroclimatic conditions with respect to oil, fats and protein contents. Cottonseed oil is routinely used for cooking and food manufacturing products. Cottonseed oil obtained after proper extraction/processing steps from crude state to refined oil in a variety of ways. Cotton crop is considered for their dual-use purpose, for fiber quality and oil production to promote health benefits in the world. Keeping in view the above facts, this review clearly demonstrated an overview about physicochemical and functional properties of cottonseed oil to promote health benefits associated with the use of this oil. The overall characteristics and all concerned health benefits of CSO will further improve their usefulness is a compact way. We have summarized a brief multi-dimensional features of CSO in all aspects up to the best of our knowledge for the end researchers who can further research in the respective aspect.

Os óleos vegetais têm propriedades físico-químicas específicas que desempenham um papel vital na dieta nutricional humana em benefício à saúde. O óleo de semente de algodão, utilizado rotineiramente no preparo e na fabricação de alimentos, é obtido através de várias espécies de sementes de algodão, famosas pela alta qualidade de sua fibra., cuja espécie mais utilizada é Gossypium hirsutum. As sementes variam, em relação ao teor do óleo, da gordura e das proteínas, de acordo com o cultivo e as diversas condições agroclimáticas. O óleo de algodão é obtido após etapas adequadas de extração das sementes e processamento do estado bruto ao refinado. Assim, a presente revisão demonstra, de maneira geral, as propriedades físico-químicas e funcionais do óleo de semente de algodão e seus benefícios à saúde humana, resumindo suas características multidimensionais. As características gerais e todos os benefícios do composto podem melhorar ainda mais se utilizadas de forma compacta, auxiliando futuros pesquisadores.

Impact of nitrogen and phosphorus fertilizers on Cry1Ac protein contents in transgenic cotton.

Application of different fertilizers to check the efficiency of expression of Bt (Bacillus thuringiensis) gene in one of the leading commercialized crops (cotton) against Lepidopteran species is of great concern. The expression of Cry protein level can be controlled by the improvement of nutrients levels. Therefore, the myth of response of Cry toxin to different combinations of NP fertilizers was explored in three Bt cotton cultivars. Combinations include three levels of nitrogen and three levels of phosphorus fertilizers. Immunostrips and Cry gene(s) specific primer based PCR (Polymerase Chain Reaction) analysis were used for the presence of Bt gene that unveiled the presence of Cry1Ac gene only. Further, the ELISA (enzyme-linked immunosorbent assay) kit was used to quantify the expression of Cry1Ac protein. Under various NP fertilizers rates, the level of toxin protein exhibited highly significant differences. The highest toxin level mean was found to be 2.3740 and 2.1732 µg/g under the treatment of N150P75 kg ha-1 combination while the lowest toxin level mean was found to be 0.9158 and 0.7641 µg/g at the N50P25 kg ha-1 level at 80 and 120 DAS (Days After Sowing), respectively. It was concluded from the research that the usage of NP fertilizers has a positive relation with the expression of Cry1Ac toxin in Bt cotton. We recommend using the N150P50 kg ha-1 level as the most economical and practicable fertilizer instead of the standard dose N100P50 kg ha-1 to get the desired level of Cry1Ac level for long lasting plant resistance (<1.5). The revised dose of fertilizer may help farmers to avoid the cross-resistance development in contradiction of insect pests.

Physicochemical features, functional characteristics, and health benefits of cottonseed oil: a review.

Vegetable oils have their specific physicochemical properties due to which they are playing vital role in human nutritional diet for health benefits. Cottonseed oil is obtained from various species of cotton seeds that are famous to be grown mainly for their fiber quality. The most prominently used specie is Gossypium hirsutum. It is obvious that the seeds of different variety of cotton vary as grown in diverse agroclimatic conditions with respect to oil, fats and protein contents. Cottonseed oil is routinely used for cooking and food manufacturing products. Cottonseed oil obtained after proper extraction/processing steps from crude state to refined oil in a variety of ways. Cotton crop is considered for their dual-use purpose, for fiber quality and oil production to promote health benefits in the world. Keeping in view the above facts, this review clearly demonstrated an overview about physicochemical and functional properties of cottonseed oil to promote health benefits associated with the use of this oil. The overall characteristics and all concerned health benefits of CSO will further improve their usefulness is a compact way. We have summarized a brief multi-dimensional features of CSO in all aspects up to the best of our knowledge for the end researchers who can further research in the respective aspect.

Molecular characterization of Ephedra species found in Pakistan.

Ephedra, also known as "ma huang", is a dioecious, drought- and frost-resistant, perennial, evergreen shrub with compelling medicinal value. The genus is represented by 42 species around the world, 9 of which were provisionally reported from Pakistan. Species of the genus have a controversial taxonomy due to their overlapping morphological features. Conventional tools alone are not sufficient for characterizing the species. The objective of present study was to assess the genetic variability present in different biotypes of Ephedra growing in Pakistan using molecular markers. A total of six genotypes collected from diverse geographic zones of Pakistan were used. The DNA of all genotypes was amplified using nine randomly amplified polymorphic DNA (RAPD) primers to study genetic variability at the molecular level. The dissimilarity coefficient matrix based on the data of 9 RAPD primers was used to construct a dendrogram which was then used to group the genotypes in clusters. Based on the dendrogram and dissimilarity coefficient matrix, the RAPD markers used here revealed a moderate to high level of genetic polymorphism (6 to 49%) among the genotypes. It was found that the collection of genotype accessions from Swat Valley in northwestern Pakistan was most distantly related to the other five collections. More molecular markers including functional genes and ribosomal spacer regions are suggested to find a better estimate of the genetic diversity present in Ephedra growing in Pakistan. The information provided here is useful for identifying valuable Ephedra variants which will be used for medicinal purposes and earning foreign currency.

Molecular characterization of Ephedra species found in Pakistan

Ephedra, also known as “ma huang”, is a dioecious, drought- and frost-resistant, perennial, evergreen shrub with compelling medicinal value. The genus is represented by 42 species around the world, 9 of which were provisionally reported from Pakistan. Species of the genus have a controversial taxonomy due to their overlapping morphological features. Conventional tools alone are not sufficient for characterizing the species. The objective of present study was to assess the genetic variability present in different biotypes of Ephedra growing in Pakistan using molecular markers. A total of six genotypes collected from diverse geographic zones of Pakistan were used. The DNA of all genotypes was amplified using nine randomly amplified polymorphic DNA (RAPD) primers to study genetic variability at the molecular level. The dissimilarity coefficient matrix based on the data of 9 RAPD primers was used to construct a dendrogram which was then used to group the genotypes in clusters. Based on the dendrogram and dissimilarity coefficient matrix, the RAPD markers used here revealed a moderate to high level of genetic polymorphism (6 to 49%) among the genotypes. It was found that the collection of genotype accessions from Swat Valley in northwestern Pakistan was most distantly related to the other five collections. More molecular markers including functional genes and ribosomal spacer regions are suggested to find a better estimate of the genetic diversity present in Ephedra growing in Pakistan. The information provided here is useful for identifying valuable Ephedra variants which will be used for medicinal purposes and earning foreign currency.

Comparison of aztreonam against other antibiotics used in urinary tract infections.

BACKGROUND: Urinary tract infection is a very common problem in adults as well as in children. There is always need for the right antibiotic to be chosen for treatment. METHODS: This study was conducted at Microbiology section in Pathology department of Ayub Medical College, Abbottabad, Pakistan. In this study the aztronam which is only effective against gram negative bacilli has been compared with other conventionally used antibiotics, ampicillin, cotrimoxazole, minocyclin, pipemedic acid, nalidixic acid, norfloxacin, ciprofloxacin, gentamicin and ceftriaxon. RESULTS: Total gram negative isolates were 342. Out of this 76.6% (262) were E. coli, klebsiella pneumoniae 14.3% (49), proteus species 5.2%(18) and pseudomonas aeruginosa were 3.8% (13). The aztreonam showed 78% sensitivity against gram negative bacilli which is better than norfloxacin which showed 62.2% sensitivity.