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Afroz A.,Quaid-i-Azam University | Afroz A.,National Institute for Genomics and Advanced Biotechnology NIGAB | Chaudhry Z.,Hazara University | Rashid U.,National Institute for Genomics and Advanced Biotechnology NIGAB | And 4 more authors.
Plant Cell, Tissue and Organ Culture | Year: 2011

To enhance bacterial wilt resistance in tomato plants and simplify the protocol of Agrobacterium tumefaciens mediated gene transfer, parameters affecting transformation efficiency in tomato have been optimized. A. tumefaciens strain EHA101, harboring a recombinant binary expression vector pTCL5 containing the Xa21 gene under the control of the CaMV 35S promoter was used for transformation. Five cultivars of tomato (Rio Grande, Roma, Pusa Ruby Pant Bahr and Avinash) were tested for transformation. Transformation efficiency was highly dependent on preculture of the explants with acetosyringone, acetosyringone in co-cultivation media, shoot regeneration medium and pre-selection after co-cultivation without selective agent. One week of pre-selection following selection along with 400 μM acetosyringone resulted in 92.3% transient GUS expression efficiency in Rio Grande followed by 90.3% in Avinash. The presence and integration of the Xa21 gene in putative transgenic plants was confirmed by polymerase chain reaction (PCR) and Southern blot analyses with 4.5-42.12% PCR-positive shoots were obtained for Xa21 and hygromycin genes, respectively. Transgenic plants of the all lines showed resistance to bacterial wilt. T1 plants (resulting from self-pollination of transgenic plants) tested against Pseudomonas solanacearum inoculation in glasshouse, showed Mendelian segregation. © 2010 Springer Science+Business Media B.V.


Akhtar S.,National Institute for Genomics and Advanced Biotechnology NIGAB | Akhtar S.,Pmas Arid Agriculture University | Akhtar S.,International Islamic University, Islamabad | Shahzad A.,National Institute for Genomics and Advanced Biotechnology NIGAB | And 4 more authors.
Pakistan Journal of Agricultural Sciences | Year: 2014

Groundnut is prone to iron (Fe) deficiency, particularly in calcareous soils, which severely affect growth and productivity. The soils of major groundnut producing areas of Pakistan are calcareous in nature, resulting in limited yield. Hydroponics experiments were performed aiming at the selection of groundnut genotypes with better ability to grow in calcareous soils. For that purpose, 20 locally grown genotypes were screened for their tolerance in terms of morpho-physiological parameters against iron (Fe) deficiency through hydroponics experiments. Various morpho-physiological parameters revealed BARI- 2000 and 96CG009 as Fe deficiency tolerant genotypes, whereas BARD-699 and ICGS17 as Fe deficiency sensitive genotypes. The Fe-reduction capacity of roots of these genotypes was also estimated at 2, 4, 6 and 8 day interval. The results depicted the highest Fe-reduction capacity at day 4 among all the genotypes, which decreased at day 6 and 8. At day 2, Fereduction capacity of 96CG009 was found higher representing its early response to Fe deficiency. Fe-reduction capacity of BARI-2000 was the highest indicating its tolerance to Fe deficiency, whereas BARD-699 was sensitive to Fe deficiency. The genetic differences among groundnut genotypes were analyzed using thirty SSR markers. These markers amicably differentiated all the genotypes resulting in three main clusters. The phylogenetic analyses based on SSR markers data revealed that Fe deficiency tolerant genotypes tended to cluster together. This implicates that molecular markers can be used for selection of groundnut genotypes with better traits. © 2014 University of Agriculture. All rights reserved.


Qamar M.,University of AJK | Qamar M.,Pakistan National Agricultural Research Center | Dilnawaz Ahmad S.,University of AJK | Ashiq Rabbani M.,Plant Genetic Resources Institute PGRI | And 2 more authors.
Pakistan Journal of Botany | Year: 2014

Stripe and leaf rusts are the major constraints to bread wheat production in Pakistan. Molecular markers were used to investigate the presence of leaf rust and stripe rust resistance gene cluster Lr34/Yr18 and stem rust resistance gene Sr2 in 52 Pakistani bread wheat cultivars/lines. PCR amplification of DNA fragments using DNA marker csLV-34 showed that 13 of the studied cultivars/lines, namely '03FJ26', 'NR 337', 'NR 339' 'NR 347', 'NR 350', 'Manthar', 'Margalla 99', 'Iqbal 2000', 'Saleem 2000', 'Wafaq 2001', 'Marwat 2001', 'Pirsabak 2004' and 'Fareed 2006' carry leaf rust and stripe rust resistance genes Lr34/Yr18. Stem rust resistance gene Sr2 was observed in 36 Pakistani spring wheat cultivars/lines using stm560.3tgag marker. The slow rusting gene Sr2 needs to be combined with additional stem rust resistance genes to establish durable resistance against Ug99 in modern wheat cultivars. Low frequency of Lr34/Yr18 was found in Pakistani wheats. This gene cluster needs to be incorporated into Pakistani wheats for durable rust resistance.


Ashraf S.,PARC Institute of Advance Studies in Agriculture | Shahzad A.,National Institute for Genomics and Advanced Biotechnology NIGAB | Shahzad A.,PARC Institute of Advance Studies in Agriculture | Karamat F.,PARC Institute of Advance Studies in Agriculture | And 4 more authors.
Journal of Animal and Plant Sciences | Year: 2015

Drought is a major limiting factor affecting productivity. Wheat is a major crop and staple food in Pakistan. Genetic linkage map construction based on linked DNA markers spanning whole wheat genome and subsequently QTL mapping for drought tolerance is a way forward to enhance breeder’s ability for effective selection. An F 8 population derived from the cross of OPATA x SH-349 has been used. An experiment was conducted at germination stage under controlled conditions. The drought was induced by15% PEG nutrient solution in acid washed sand medium. Microsatellite DNA markers data were used for linkage maps construction. Morphological data under drought and non-stressed conditions along with linkage maps data were used for QTL mapping. The results of QTL analysis using single marker analysis showed 14 SSR markers linked to QTLs for five traits in both drought and control condition. Using simple interval mapping and composite interval mapping 12 QTLs for different traits of interest were found. Implications of found QTLs and linked markers are discussed. © 2015, Pakistan Agricultural Scientists Forum. All rights reserved.


Jan S.A.,Quaid-i-Azam University | Shinwari Z.K.,Quaid-i-Azam University | Shah S.H.,Allama Iqbal Open University | Shahzad A.,National Institute for Genomics and Advanced Biotechnology NIGAB | And 2 more authors.
Romanian Biotechnological Letters | Year: 2016

In-planta transformation is an efficient, quick and tissue culture independent system for crop plants improvement. In-planta transformation is a useful system for those plants that lack tissue culture and regeneration system. Therefore, the direct in-planta methods are commonly used for the transformation of many important genes into these plant species. The two most common Agrobacterium mediated in-planta methods such as floral dip and vacuum infiltration have been successfully used by many researchers in both dicot and monocot plants. The main advantages of in-planta transformation are to produce large number of transgenic plants and accumulation of high concentration of total soluble protein in short time. The production of chimera plants and lack of stable transgene expression at T1 or after T1 stage are some of the problems to these methods that need further improvements. The present review mainly focuses on in-planta transformation protocols that were successfully used for many agronomical important crop plants and to describe the various advantages and disadvantages of in-planta transformation methods in different crop species. © 2016 University of Bucharest.


Rana A.,National Institute for Genomics and Advanced Biotechnology NIGAB | Rana A.,National University of Sciences and Technology | Ali G.M.,National Institute for Genomics and Advanced Biotechnology NIGAB | Ali S.,National Institute for Genomics and Advanced Biotechnology NIGAB | And 4 more authors.
Journal of Cancer Research and Therapeutics | Year: 2013

Leukemia is a many-sided molecular disorder that arises because of over expression of oncogenes, suppression of tumor suppressor genes, and chromosomal translocations. These chromosomal rearrangements are nonetheless among the many determinants that underlie transformation of cells from normal to a cancerous phenotype and predispose cells to refractoriness against interventions by reduced drug influx and substantial drug efflux. This review unfolds current understanding of BCR-ABL1 (break point cluster region-c-abl oncogene 1, non-receptor tyrosine kinase) signaling with a focus on apoptotic suppressive mechanisms and alternative approaches to chronic myeloid leukemia therapy.


Abbas Z.,Pakistan National Agricultural Research Center | Abbas Z.,National Institute for Genomics and Advanced Biotechnology NIGAB | Abbas Z.,National Institute for Biotechnology and Genetic Engineering NIBGE | Zafar Y.,Pakistan Atomic Energy Commission PAEC | And 2 more authors.
International Journal of Agriculture and Biology | Year: 2013

Insects have natural potential to develop resistance against chemical insecticides. Several resistance strategies have been suggested including biopesticides and use of two dissimilar toxins. Advances in molecular biology techniques have now allowed construction of chimeric proteins to delay the development of resistance in insect population, but still there are chances of developing resistance in insect population against them as these fusions are based on Bacillus thuringiensis (Bt) genes only, which have some homology in their amino acid sequences, having same mode of action and derived from same bacterial origin. In the present study ω-ACTX-Hv1a toxin gene (Hvt) as an insect calcium channel antagonist is fused with Bt cry1Ac to combine both strategies (biopesticides and two dissimilar toxins) and delay the resistance in insect population. The recombinant protein has been successfully expressed in prokaryotic system and was detected by SDS PAGE. Topical application of the 1.0 pmol purified recombinant protein to the thoracic region paralyzed and immobilized the Helicoverpa armigera and Spodoptera littoralis larvae within 2 h. 100% mortality was observed in insects after 24 h. The LD50 was found to be 4 and 2 pmol per gram of body weight for H. armigera and S. littoralis larvae, respectively. The present study clearly indicates that this recombinant protein is highly effective against agronomical important lepidopteron insects and is an excellent candidate for use as a biopesticides or expressed heterogeneously in agricultural crops to provide long lasting resistance to insect attacks. © 2013 Friends Science Publishers.


Rana A.,National Institute for Genomics and Advanced Biotechnology NIGAB | Rana A.,National University of Sciences and Technology | Ali G.M.,National Institute for Genomics and Advanced Biotechnology NIGAB | Ali S.,National Institute for Genomics and Advanced Biotechnology NIGAB | And 5 more authors.
Journal of Cancer Research and Therapeutics | Year: 2013

Increasing sophisticated information suggests that cancer cells express constitutively active oncogenic kinases such as breakpoint cluster region- c-abl oncogene 1, non-receptor tyrosine kinase (BCR-ABL1) that promote carcinogenesis independent of extrinsic growth factors. It is a well-established fact that through the aberrant activation of BCR-ABL1 signal transduction cascade, the perception of cellular growth signals becomes disconnected from the processes promoting cell growth, and this underlies the pathophysiology of leukemia. In this particular review we discuss the oncogenes and tumor suppressors comprising the regulatory network upstream and downstream of BCR-ABL1 and dismantle how derailed BCR-ABL1 signaling provides cell a selective growth advantage. Besides, we discuss why activation of BCR-ABL1, as an outcome of distinct oncogenic events, results in miscellaneous clinical outcomes, and how the intricacy of the BCR-ABL1 signaling network might dictate therapeutic approaches. In this review, our current comprehension of BCR-ABL1 signaling will be summarized.


PubMed | National Institute for Genomics and Advanced Biotechnology NIGAB
Type: Journal Article | Journal: Asian Pacific journal of cancer prevention : APJCP | Year: 2014

In-depth analysis of how TRAIL signals through death receptors to induce apoptosis in cancer cells using high throughput technologies has added new layers of knowledge. However, the wealth of information has also highlighted the fact that TRAIL induced apoptosis may be impaired as evidenced by experimental findings obtained from TRAIL resistant cancer cell lines. Overwhelmingly, increasing understanding of TRAIL mediated apoptosis has helped in identifying synthetic and natural compounds which can restore TRAIL induced apoptosis via functionalization of either extrinsic or intrinsic pathways. Increasingly it is being realized that biologically active phytochemicals modulate TRAIL induced apoptosis, as evidenced by cell-based studies. In this review we have attempted to provide an overview of how different phytonutrients have shown efficacy in restoring apoptosis in TRAIL resistant cancer cells. We partition this review into how the TRAIL mediated signaling landscape has broadened over the years and how TRAIL induced signaling machinery crosstalks with autophagic protein networks. Subsequently, we provide a generalized view of considerable biological activity of coumarins against a wide range of cancer cell lines and how coumarins (psoralidin and esculetin) isolated from natural sources have improved TRAIL induced apoptosis in resistant cancer cells. We summarize recent updates on piperlongumine, phenethyl isothiocyanate and luteolin induced activation of TRAIL mediated apoptosis. The data obtained from pre-clinical studies will be helpful in translation of information from benchtop to the bedside.

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