More than 50 countries around the globe cultivate cotton on a large scale. It is a major cash crop of Pakistan and is considered “white gold” because it is highly important to the economy of Pakistan. In addition to its importance, cotton cultivation faces several problems, such as insect pests, weeds, and viruses. In the past, insects have been controlled by insecticides, but this method caused a severe loss to the economy. However, conventional breeding methods have provided considerable breakthroughs in the improvement of cotton, but it also has several limitations. In comparison with conventional methods, biotechnology has the potential to create genetically modified plants that are environmentally safe and economically viable. In this study, a local cotton variety VH 289 was transformed with two Bt genes (Cry1Ac and Cry2A) and a herbicide resistant gene (cp4 EPSPS) using the Agrobacterium mediated transformation method. The constitutive CaMV 35S promoter was attached to the genes taken from Bacillus thuringiensis (Bt) and to an herbicide resistant gene during cloning, and this promoter was used for the expression of the genes in cotton plants. This construct was used to develop the Glyphosate Tolerance Gene (GTGene) for herbicide tolerance and insecticidal gene (Cry1Ac and Cry2A) for insect tolerance in the cotton variety VH 289. The transgenic cotton variety performed 85% better compared with the non-transgenic variety. The study results suggest that farmers should use the transgenic cotton variety for general cultivation to improve the production of cotton.
BackgroundCotton yield has been badly affected by different insects and weed competition. In Past Application of multiple chemicals is required to manage insects and weed control was achieved by different conventional means, such as hand weeding, crop rotation and polyculture, because no synthetic chemicals were available. The control methods shifted towards high input and target-oriented methods after the discovery of synthetic herbicide in the 1930s. To utilise the transgenic approach, cotton plants expressing the codon-optimised CEMB GTGene were produced in the present study.ResultsLocal cotton variety CEMB-02 containing Cry1Ac and Cry2A in single cassette was transformed by synthetic codon-optimised 5-enolpyruvylshikimate-3-phosphate synthase gene cloned into pCAMBIA 1301 vector under 35S promoter with Agrobacterium tumifaciens. Putative transgenic plants were screened in MS medium containing 120 µmol/L glyphosate. Integration and expression of the gene were evaluated by PCR from genomic DNA and ELISA from protein. A 1.4-kb PCR product for Glyphosate and 167-bp product for Cry2A were obtained by amplification through gene specific primers. Expression level of Glyphosate and Bt proteins in two transgenic lines were recorded to be 0.362, 0.325 µg/g leaf and 0.390, 0.300 µg/g leaf respectively. FISH analysis of transgenic lines demonstrates the presence of one and two copy no. of Cp4 EPSPS transgene respectively. Efficacy of the transgene Cp4 EPSPS was further evaluated by Glyphosate spray (41 %) assay at 1900 ml/acre and insect bioassay which shows 100 %mortality of insect feeding on transgenic lines as compared to control.ConclusionThe present study shows that the transgenic lines produced in this study were resistant not only to insects but also equally good against 1900 ml/acre field spray concentration of glyphosate.Electronic supplementary materialThe online version of this article (doi:10.1186/s13104-015-1397-0) contains supplementary material, which is available to authorized users.
Cotton is a premium source of natural fiber and is considered Bwhite gold^in the textile industry. Fiber length, strength and fineness are the key considerations for the industry. Longer fibers are machine friendly because they are easily spinnable. Recent advancements in genetic engineering, including the development of DNA markers and quantitative trait loci (QTLs), together with genome sequencing and gene expression profiling, have provided new avenues for improving fiber production and quality. In plants, sucrose synthase (SUS) is the key enzyme that catalyzes the reversible cleavage of sucrose and uridine diphosphate (UDP) into fructose and UDP-glucose. Sucrose is the main mobile sugar in plants moving from source to sink. It regulates resource partitioning between active sinks, especially in cotton embryos and fibers, and therefore is directly involved in determining fiber yield and seed quality. SUS actively takes part in regulating the competition for nutrients among sink tissues through balancing osmotic potentials by providing hexoses and an efficient supply of UDP-glucose the substrate for cellulose synthase. Cotton transformation has been used to improve fiber characteristics by altering cell wall properties through the manipulation of expression of fiber genes. Overexpression of the SUS gene from natural or synthetic origins in cotton can be an excellent way to solve potential problems associated with poor fiber length and other fiber quality traits. Increased SUS activity can result in more hexoses, increasing the osmotic potential and thereby driving the water influx that creates high turgor pressure in fiber cells resulting in enhanced fiber elongation. Moreover, increased SUS gene transcript levels in vegetative tissues of the plant will elevate seedling biomass and seed number. Fiber length and seed number both contribute towards final yield and the SUS genes as key regulators of sink strength in cotton perform this dual function that is directly related to cotton productivity. Hence manipulation of the SUS gene family is considered a promising approach to improve cotton fiber yield and quality. This review focuses on the biochemical and physiological roles of the SUS genes and there value for cotton fiber improvement.
The study aims to improve fiber traits of local cotton cultivar through genetic transformation of sucrose synthase ( SuS ) gene in cotton. Sucrose synthase (SuS) is an important factor that is involved in the conversion of sucrose to fructose and UDP-glucose, which are essential for the synthesis of cell wall cellulose. In the current study, we expressed a synthetic SuS gene in cotton plants under the control of a CaMV35S promoter. Amplification of an 813-bp fragment using gene-specific primers confirmed the successful introduction of SuS gene into the genome of cotton variety CEMB-00. High SuS mRNA expression was observed in two transgenic cotton plants, MA0023 and MA0034, when compared to the expression in two other transgenic cotton plants, MA0035 and MA0038. Experiments showed that SuS mRNA expression was positively correlated with SuS activity at the vegetative (54%) and reproductive stages (40%). Furthermore, location of transgene was found to be at chromosome no. 9 in the form of single insertion, while no signal was evident in non-transgenic control cotton plant when evaluated through fluorescent in situ hybridization and karyotyping analysis. Fiber analyses of the transgenic cotton plants showed increases of 11.7% fiber length, 18.65% fiber strength, and up to 5% cellulose contents. An improvement in the micronaire value of 4.21 was also observed in the MA0038 transgenic cotton line. Scanning electron microscopy (SEM) revealed that the fibers of the SuS transgenic cotton plants were highly spiral with a greater number of twists per unit length than the fibers of the non-transgenic control plants. These results determined that SuS gene expression influenced cotton fiber structure and quality, suggesting that SuS gene has great potential for cotton fiber quality improvement.
Newcastle disease (ND) is a viral disease that causes labored breathing, periorbital oedema, and ataxia in the majority of avian species. The available vaccines against Newcastle disease virus (NDV) are limited, owing to their low reactivity and multiple dosage requirements. Plant-based machinery provides an attractive and safe system for vaccine production. In the current study, we attempted to express fusion (F) and hemagglutinin-neuraminidase (HN) proteins (the protective antigens against NDV) under constitutive 35S and seed-specific Zein promoters, respectively. Almost 2-7.1-fold higher expression of F gene mRNA in transgenic corn leaves and 8-28-fold higher expression of HN gene mRNA in transgenic corn seeds were observed, when the expression was analyzed by real-time PCR on a relative basis as compared to non-transgenic control plant material (Leaves and seeds). Similarly, 1.66 µg/ml of F protein in corn leaves, i.e., 0.5% of total soluble protein, and 2.4 µg/ml of HN protein in corn seed, i.e., 0.8% of total seed protein, were found when calculated through ELISA. Similar levels of immunological response were generated in chicks immunized through injection of E. coli-produced pET F and pET HN protein as in chickens orally fed leaves and seeds of maize with expressed immunogenic protein. Moreover, the detection of anti-NDV antibodies in the sera of chickens that were fed maize with immunogenic protein, and the absence of these antibodies in chickens fed a normal diet, confirmed the specificity of the antibodies generated through feeding, and demonstrated the potential of utilizing plants for producing more vaccine doses, vaccine generation at higher levels and against other infectious diseases.
Gout is a type of inflammatory arthritis with drastically increasing incidence since last twenty years. Increased serum uric acid levels and deposition of urate crystals in joints and soft tissues are known to stir-up the symptoms of gout. Patient's quality of life (Qol) suffers markedly due to inflammation, pain, restricted mobility, health economic burden and suboptimal therapeutics associated with gout. Related articles published between 2007 and 2018 were reviewed with the help of different databases including PubMed, Springer link, Medline, Google scholar and Science direct. To ensure the accuracy and credibility of data, articles published only in indexed journals were considered. Gout is a common metabolic disorder that is provoked by increased level of serum uric acid. Presence of other comorbidities i.e. hypertension, diabetes and cardiac diseases precipitate the rate of mortality. Number of gout patients is shown to increase in developed countries whereas developing countries remain at high risk category. The enormous increase in gout is due to chronicity, lack of awareness among patients, poor diagnosis, suboptimal treatment and economic burden. Risk factors associated with gout are well identified and pathophysiology is well-established hence, measures can be taken to improve diagnosis and to ensure cost effective options for the treatment. To curtail the global health burden and to improve patient quality of life there is need: to reduce exposure to risk factors, to promote awareness, to institute robust prophylactic measures, to modify lifestyle, to establish signs for early diagnosis and to develop optimized treatment strategies and policies.
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