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Background: Empirical inculcation is vital for producing quality research professionals and upon realizing the increasing demand of emerging scientific research professionals, National Academy of Young Scientist (NAYS) launched its science for youth (S4Y) program in Pakistan with its emphasis on empiricism. Methods: S4Y team was galvanized to undertake this task and different scientific activities were organized by them in different schools and colleges of Pakistan during 2011 _ 2012. Scientific activities that were arranged included seminars on diverse scientific subjects and career counseling sessions. Results: Response of students showed that the implementation of this program can contribute in achieving the target of producing qualified skilled research professionals and upturning the Pakistan literacy rate. Experimental activities were also planned but its implementation was restricted due to financial strains. 39 activities were conducted in 2012 in the schools and colleges of all the provinces of Pakistan on the importance of science in the world, scientific discoveries, and career counseling. Conclusion: These activities at school and college level were appreciated by the students and staff members and proved very fruitful for defining the scientific fields and goals of scientific studies for Pakistani youth. Implementation of experimental activities in future will help students in getting awareness about the ways through which science explores this world. [ABSTRACT FROM AUTHOR]

Genome editing technology has become part and parcel of biotechnological approach to understand the genetic basis of different biological processes in an organism and for its application in different fields. The basic need of world food production demands a highly variable population of plants with maximum diversity of agronomic traits of interest. Genome editing is unique in the sense that it provides a direct approach to generate targeted variability. The worldwide availability of genome‐edited crop varieties in the global market is approaching, whereas the decision to deal with genome‐edited (GE) crops differently from genetically transformed crop varieties is under discussion all around the world. GE may add allochthonous or autochthonous sequences into the genome much like traditional transgenesis or cisgenesis. Considering this fact, the United States declared genome‐edited crops as conventional. Many countries such as Japan, UK, China, and Brazil have already legislated their GE products and their products are near to get entry to the market. In Pakistan, academic and research‐based scientific institutions have been working on genome‐edited crops by using CRISPR (clustered regularly interspaced short palindromic repeats) Cas technology. As being a signatory of the Cartagena protocol, every case of GM (Genetically modified) or GE (Genetically Engineered) should be passed through the Technical Advisory Committee which will be decided on case‐to‐case bases depending upon its nature of random or targeted mutation. Pakistan Environmental Protection Act, Intellectual Property Organization of Pakistan Act, Seed amendment act, and PBR (The Plant Breeders' Rights Act) are the major constitutional acts employed and working for the regulations of GM alongside GE crops. Now, discussions of scientists, academicians and officials from regulatory body are in process for the design of GE crop commercialization policy in Pakistan. The CRISPR‐Cas9 system is now the most in‐demand piece of technology and has become an indispensable component of research and development in the field of life sciences. [ABSTRACT FROM AUTHOR]

Newcastle disease virus (NDV) is a devastating virus for the poultry industry, resulting in huge economic losses due to high morbidity and mortality. Commercial vaccines are available, but they are limited because of their high cost, difficult storage and administration system, and poor protection against local rising strains. Microalgal chloroplasts can be used to develop recombinant subunit vaccines having the advantages of scalable growth, quick transformation, and uniform expression levels. These vaccines can be produced on a large scale at an economical cost and are expected to be used to induce antigen-specific responses for a range of human and animal diseases. This study was aimed to develop an algae-based vaccine against NDV and to determine its effect on the immune response in chickens. The NDV HN gene was integrated into the chloroplast of Chlamydomonas reinhardtii to obtain stable and high expression of antigenic HN gene, which was found to be ~ 1000 fold higher than C. reinhardtii TN72 cell lines. The grown algal cell lines containing HN gene were fed to the chickens and their immune responses were evaluated in their blood through real time PCR (RT-PCR) analysis. The expression analysis showed a significant upregulation in the expression of Interleukin-6, Interleukins-8, and Interferon-γ at 16th and 26th day of vaccination with transgenic alga. The success of the study proved that transgenic algae-based vaccines can be used to protect poultry from NDV infection. [ABSTRACT FROM AUTHOR]

Premature leaf senescence negatively influences the physiology and yield of cotton plants. The conserved IDLNL sequence in the C-terminal region of AGL42 MADS-box determines its repressor potential for the down regulation of senescence-related genes. To determine the delay in premature leaf senescence, Arabidopsis AGL42 gene was overexpressed in cotton plants. The absolute quantification of transgenic cotton plants revealed higher mRNA expression of AGL42 compared to that of the non-transgenic control. The spatial expression of GUS fused with AGL42 and the mRNA level was highest in the petals, abscission zone (flower and bud), 8 days post anthesis (DPA) fiber, fresh mature leaves, and senescenced leaves. The mRNA levels of different NAC senescence-promoting genes were significantly downregulated in AGL42 transgenic cotton lines than those in the non-transgenic control. The photosynthetic rate and chlorophyll content were higher in AGL42 transgenic cotton lines than those in the non-transgenic control. Fluorescence in situ hybridization of the AG3 transgenic cotton line revealed a fluorescent signal on chromosome 1 in the hemizygous form. Moreover, the average number of bolls in the transgenic cotton lines was significantly higher than that in the non-transgenic control because of the higher retention of floral buds and squares, which has the potential to improve cotton fiber yield. [ABSTRACT FROM AUTHOR]

Whitefly infestation of cotton crop imparts enormous damage to cotton yield by severely affecting plant health, vigour and transmitting Cotton Leaf Curl Virus (CLCuV). Genetic modification of cotton helps to overcome both the direct whitefly infestation as well as CLCuV based cotton yield losses. We have constitutively overexpressed asparaginase (ZmASN) gene in Gossypium hirsutum to overcome the cotton yield losses imparted by whitefly infestation. We achieved 2.54% transformation efficiency in CIM-482 by Agrobacterium-mediated shoot apex transformation method. The relative qRT-PCR revealed 40-fold higher transcripts of asparaginase in transgenic cotton line vs. non-transgenic cotton lines. Metabolic analysis showed higher contents of aspartic acid and glutamic acid in seeds and phloem sap of the transgenic cotton lines. Phenotypically, the transgenic cotton lines showed vigorous growth and height, greater number of bolls, and yield. Among six representative transgenic cotton lines, line 14 had higher photosynthetic rate, stomatal conductance, smooth fiber surface, increased fiber convolutions (SEM analysis) and 95% whitefly mortality as compared to non-transgenic cotton line. The gene integration analysis by fluorescence in situ hybridization showed single copy gene integration at chromosome number 1. Collectively, asparaginase gene demonstrated potential to control whitefly infestation, post-infestation damages and improve cotton plant health and yield: a pre-requisite for farmer's community. [ABSTRACT FROM AUTHOR]

Cotton leaf curl virus disease (CLCuD) is caused by a suite of whitefly-transmitted begomovirus species and strains, resulting in extensive losses annually in India and Pakistan. RNA-interference (RNAi) is a proven technology used for knockdown of gene expression in higher organisms and viruses. In this study, a small interfering RNA (siRNA) construct was designed to target the AC1 gene of Cotton leaf curl Kokhran virus-Burewala (CLCuKoV-Bu) and the bC1 gene and satellite conserved region of the Cotton leaf curl Multan betasatellite (CLCuMB). The AC1 gene and CLCuMB coding and non-coding regions function in replication initiation and suppression of the plant host defense pathway, respectively. The construct, Vb, was transformed into cotton plants using the Agrobacterium-mediated embryo shoot apex cut method. Results from fluorescence in situ hybridization and karyotyping assays indicated that six of the 11 T1 plants harbored a single copy of the Vβ transgene. Transgenic cotton plants and non-transgenic (susceptible) test plants included as the positive control were challenge-inoculated using the viruliferous whitefly vector to transmit the CLCuKoV-Bu/CLCuMB complex. Among the test plants, plant Vβ-6 was asymptomatic, had the lowest amount of detectable virus, and harbored a single copy of the transgene on chromosome six. Absence of characteristic leaf curl symptom development in transgenic Vβ-6 cotton plants, and significantly reduced begomoviral-betasatellite accumulation based on real-time polymerase chain reaction, indicated the successful knockdown of CLCuKoV-Bu and CLCuMB expression, resulting in leaf curl resistant plants. [ABSTRACT FROM AUTHOR]

Background: Pesticides and insecticides are very toxic by design and they are used world over to protect the crops from insects. Although, they are good for better crops production but they have serious effects on environment and human health. Exposure to pesticides and insecticides is one of the most important occupational health risks in developing countries, like Pakistan. Methods: A survey was designed to know the awareness level about insecticides and pesticides toxicity and spray procedure in Punjab, Pakistan. Results: Majority of respondents was from village, in the age group of 25-34, with spray experience of 0-5 years and did not have any formal education. Mostly learned about spray procedure themselves orfrom family and friends and none of them had training from government institutes. Most of the respondents did not use any protection equipment's and also had different health problems during spray process. They also reported deaths due to acute poisoning of spray personals. Conclusion: The results clearly indicate that there is great need to aware people about toxicity of pesticides and insecticides as well as educate them about personal, family and environmental protection. Government and nongovernment organizations as well as print and electronic media can play an important role in this regards. There is need of urgent steps to stop acute and chronic poisoning of poor pesticide handlers and make this profession safer. [ABSTRACT FROM AUTHOR]