With the advent of genetic manipulation techniques, it has become possible to clone and insert gene into the genome of crop plants to confer resistance to insects and pests. Resistance to insects has been demonstrating in transgenic plants either by triggering defense system of plants or by expressing heterologous cry genes for delta-endotoxins from Bacillus thuringiensis. In the present study, synthetic cry1Ab gene was developed with optimized chloroplast preferred codons and is expressed in tobacco plastid genome called plastome, following chloroplast transformation strategy, which is environment friendly technique to minimize out-crossing of transgenes to related weeds and crops. In addition, due to high polyploidy of plastid genome transformation of chloroplast permits the introduction of thousands of copies of foreign genes per plant cell, leading to extraordinarily high levels of foreign protein expression. The chloroplast transformation technology aims to insert stably into the plastome through homologous recombination into pre-decided position. To characterize the synthetic cry1Ab gene, chloroplast transformation vectors were developed and bombarded to the leaf cells of tobacco plants maintained under aseptic conditions. After bombardment, the drug resistant shoots were selected and regenerated on drug containing regeneration medium. Homoplasmic shoots were recovered after successive rounds of selection and regeneration. Proliferated plants were subjected to genomic DNA analysis by using polymerase chain reaction (PCR) technique where cry1Ab gene-specific primers were used. PCR positive plants were subjected to protein analysis, and functionally expressed proteins were detected using Immuno-Strips specific for cry1Ab/Ac gene products. Transgenic plants carrying cry1Ab gene were found expressing Bt toxins confirming that engineered gene could be expressed in other plants as well.

Mosquitoes represent the major vectors involved in the spread of deadly diseases like Malaria, Lymphatic filariasis,Chikungunyaand arboviruses like dengue virus and Zika virus. These diseases have emerged with the rise of urbanization and the use of chemical insecticides around the world. The insecticides such as DDT (Dichlorodiphenyltrichloroethane), organophosphates, carbamates, and organochlorineshave caused resistance in mosquitoes against typical chemical control methodsthrough excessive use over the years. Different biological control methods aresustainable, eco-friendly and target major diseases spreading mosquito's species. This review outlines major biological control methods being used ortested in different parts of the world. Some of these are the use of plant extract, Wolbachiaspp, larvivorous fishes, insecticidal bacterial spp, predator mosquitoes and predator copepods.Limitations associated with each method are also discussed.All these methods will givea better understanding of developing an integrated approach for effective mosquitoes control that will reduce dependency on insecticide based approaches.

The authors declare that there is no conflict of interest regarding the publication of this manuscript.

Context: In the last few decades an ample array of molecular techniques has been introduced to obtain new disposition for the classification of Trichoderma species. Today the concern of scientists is either in the direction of gene targeting or ribotyping, the newest fingerprinting tool for genomic DNA that contain all or part of the genes coding for 18S rRNA in eukaryotes.Objectives: To take advantage of advanced molecular techniques for phylogenetic analysis of indigenous isolates of Trichoderma to comprehend our knowledge of this genus by supplementing the phenotypic identification. Materials and Methods:Genomic DNA of twenty four isolates of Trichoderma species (T. harzianum, T. hamatum, T. koningii and T. pseudokoningii) were extracted by CTAB method and indicated band of ~15Kb on 0.8% agarose gel. Quality of DNA was determined by obtaining absorbance ratio (260/280) in the range of 1.7-1.9. Restriction fragment length polymorphism (RFLP) analyses were performed by using two restriction endonulease enzymes i.e., BamHI and HindIII. The BamHI represented results in the range of 500bp-750bp. 18S rRNA gene targeting was further carried out through optimization in ribotyping analysis. Results:The DNA bands of 24 isolates of Trichoderma species were compared with marker DNA bands and indicated the presence of genomic DNA intact band of ~15Kb. The ratio of absorbance 260/280nm (1.8-1.9 for pure DNA preparations) provided an estimate of the purity of the DNA RFLP analysis, along with the negative control of twenty four different isolates of Trichoderma species subjected to restriction using BamHI enzyme. The rRNA gene amplified band was observed at 600bp in the case of T. hamatum isolate (S. cumini stem bark, FCBP accession number 769) while in remaining isolates bands were in slightly smeared form. Furthermore, rRNA gene amplification conditions were optimized by altering different Tm and MgCl2 concentrations. Conclusion:The genomic DNA can serve as long term storage of information. Therefore advance molecular techniques can be used to study the variability in the genome of organism. RFLP are the initial steps for screening the genome of any organism.

Oral cavity is the main part of the digestive system which is used for the uptake of nutrients/food while microbes can also enter through the oral cavity along the uptake of food. It is seen that the composition of host saliva is very important to build the environment of the oral cavity. When the pH ofsaliva became low then the chances of microbial attacks and their adherence to the surface of teeth to form lesion increases. A complete microbial ecosystem is present in oral cavity and biofilms are produced by the microbes of normal flora to protect host from pathogenic microbes. Most of the microbes that enter oralcavity have ability to cause disease, they incorporates themselves into the normal flora and alter the biofilm community and induce autoimmunity and inflammation into the host. The personnel and materials also play crucial role in activation and inhibition of caries and lesions. Phages are gaining interest in the field of microbiology as they are less toxic and host specific for their action. Dental caries are the indication of chronic infections inside the body. By examine the oral cavity of an individual we can analyze the health status of that individual. Diseases can be controlled through better living conditions and oral hygiene, as much of the developed countries has lower risk of getting caries where as in under-developed and developing countries have greater chance to get the risk.