Historical Roots of Radicalization in Pashtun’s Society

This research article aims to trace the history of radical movements in the North-West frontier of sub-continent. Historically, radical movements have long roots in Pakhtun Society.  People recruited in different epochs from Pakhtun society branch into various freedom movements before the partition of sub-continent. Freedom movements against the Sikh, Hindu and the British lifted radical impact on Pakhtun Society before the partition of sub-continent.  Radical movements after the partition of sub-continent also established their roots in the North-West region of Pakistan. These radical movements engineered the pluralistic cultural values of Pakhtun Society. These movements have lifted radical trends in the North-West frontier of sub-continent. Pakhtuns and their cultural values were not only exposed to violence but the evolution of their culture had been disturbed.

Characterization of Defective Dna-Β Associated With Cotton Leaf Curl Disease and Their Use in Developing Enhanced Resistance.

Cotton leaf curl disease (CLCuD) has been one of the major threats to the production of cotton crop; caused by the members of genus Begomovirus of family Geminiviridae of plant viruses. Bipartite begomoviruses have two genomic components DNA A and DNA B. While, monopartite begomoviruses have a single DNA component, often associated with some satellites namely betasatellite and alphasatellite. Betasatellites depend on the helper viruses for their proliferation and in turn are required for helper virus accumulation and symptom expression. In some viruses, curtailed genomic components are produced during their replication in host cells. These components are termed as defective molecules or subgenomic particles which are known to interfere with the replication of virus by competing for host resources. Their production results in the amelioration of symptoms, delayed expression of symptoms and recovery coupled with increased resistance. Using universal betasatellite primers, both full length and defective betasatellites were detected by PCR. To determine their potential to develop resistance against homologous and heterologous viruses, defective betasatellites were cloned from the cotton samples taken from different areas of Punjab and Sindh, and characterized. Their sequence analysis revealed them as recombinant betasatellites prevailing in both provinces. A single species of Cotton leaf curl Multan betasatellite (CLCuMB) with its different recombinants was found associated with disease. Despite of high sequence homology between the betasatellite sequences from two provinces, Punjab and Sindh betasatellites were having some prominent differences particularly in the region between A-rich and SCR. This is the region thought to be involved in transreplication of betasatellite by its helper begomovirus. Sindh betasatellites encompassed the unusual sequence in this region, very rarely found in the betasatellites reported earlier. In addition to this, amino acid alignment of βC1 of these clones clearly differentiates the betasatellites of Punjab and Sindh. An analysis of subgenomic betasatellites suggested that the majority of them fall in the same size (650-700) and arose by deletion of βC1. Rolling circle amplification (RCA) techniques were used for the amplification of viral genome. RCA using Phi29 DNA polymerase amplified some non-viral sequences from infected cotton plant samples as well such as transposons, chloroplast sequences and some other sequences of unknown origin. Looped out small circular molecules of chloroplast were also detected by Southern hybridization analysis. Sequence analysis revealed that the looped out molecules contain inverted repeat sequences of chloroplast. Increased amplification of such molecules was related to virus infection and lacking in the DNA from healthy cotton samples, suggested some unrevealed mechanisms to be studied. Nicotiana species provide a good model system to understand the role of ploidy level on geminivirus-host interactions. Infectivity studies on various diploid and tetraploid Nicotiana species also established the interference of subgenomics on virus replication. The host response to begomovirus infection showed that all Nicotiana species supported begomovirus replication but only few species supported systemic infection and therefore the limitation is imposed by inhibition of virus movement. CLCuMV infection caused symptom development only in Nicotiana benthamiana while, ToLCNDV produced symptoms in three species N. benthamiana, N. tabacum and N. sylvestris. It indicates wide host range and rapid adaptability of bipartite begomoviruses as compared to monopartite ones. It was also inferred that ploidy level doesn’t play role in disease resistance or susceptibility. Despite high level of diversity in begomoviruses, only a single species of betasatellite is associated with cotton leaf curl disease. Thus, betasatellite is an attractive target for RNAi based resistance where resistance is dependent on levels of siRNA produced. βC1 is the only protein produced by betasatellite, responsible for successful infection in cotton. So in betasatellite, targeting βC1 can silence the active role of betasatellite. To enhance the resistance mechanism induced by siRNA directed against betasatellite, an amplicon-based hairpin RNAi construct was made based on defective molecule, serving dual purposes (producing defective molecules and targeting βC1). Transient infectivity analysis showed promising results which led to the stable transformation of transgene. Several lines of transgenic Nicotiana benthamiana plants were produced having RNAi construct as transgene. Infectivity of CLCuKV and CLCuMB resulted in efficient resistance response in transgenic plants. Out of 40 plants, 13 plants belonging to different transgenic lines remained symptomless, 15 plants showed mild symptoms and 12 showed delayed expression of full symptoms. Image produced through Southern hybridization showed low titre of begomovirus and no betasatellite in most plants of transgenic lines. Effect on the titre of begomovirus upon targeting βC1 further confirms the significance of the target.