International Conference on Transcriptomics July 27-29, 2015 Orlando, FL, USA

Ramarao Seriramalu is a Phd student under supervision of Associate Prof Dr.Subha Bhassu. Assoc Prof Dr. Subha Bhassu is an academician in Institute of Biological Sciences, Faculty of Science, University Malaya.

The Malaysian giant freshwater prawn, Macrobrachium rosenbergii, an economically important crustacean worldwide are being affected by Vibriosis, a disease caused by Vibrio strains such as Vibrio parahaemolyticus. M. rosenbergii possesses an innate immune system which provides defence against pathogenic agents. The information regarding the regulation of innate immune system in this species is lacking, thus its necessary in providing solutions to control and minimize the loss of production due to this bacterial disease. In this study, we performed a transcriptome profiling of M. rosenbergii hepatopancreas infected with V. parahaemolyticus using the Next Generation sequencing method (Illumina HiSeqTM2000). A total of 54,295,342 and 54,708,014 high-quality reads obtained from Vibrio-infected and control M. rosenbergii cDNA libraries. The overall de novo assembly and clustering of both reads generated 64,411 unigenes, with an average length of 698 bp. Based on BLASTX search (E-value <10-5) against NR, Swissprot, GO, COG and KEGG databases, 22,455 unigenes (34.86% of all unigenes) were annotated with gene descriptions, gene ontology terms, and metabolic pathways. The unigene differential expression analysis revealed 14,569 unigenes were differentially expressed in the infected shrimp compared to the controls. Several differentially expressed genes are involved in various animal immune functions. The large number of transcripts obtained in this study would provide valuable resources for further genomic research into freshwater prawns.

This work has been done to get more insights into prawns Immune response under viral infections

Epizootic diseases cause huge mortality and economical loses at post larvae stages in freshwater prawn aquaculture industry. These prawns seem less susceptible to viral diseases except for Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV). During viral infection in prawns, hemocytes are the primary organ that shows immunological response within the early stages of infection. We applied proteomic approaches to understand differential expression of the proteins in hemocytes during the viral disease outbreak. To aid the goal, we collected Macrobrachium rosenbergii broodstocks from the local grow out hatchery which reported the first incidence of IHHNV viral outbreak during larvae stage. Primarily, application of the OIE primer targeting 389 bp fragments of IHHNV virus were used in identification of the infected and non-infected samples of the prawn breeding line. Analysis of two-dimensional gel electrophoresis showed specific down- regulation of Arginine kinase and Sarcoplasmic calcium-binding protein and up/down- regulation of Pro-phenoloxidase1 and Hemocyanin isoforms. These proteins were validated using semi quantitative RT-PCR and gene transcripts at mRNA level. These identified proteins can be used as biomarkers, providing a powerful approach to better understanding of the immunity pathway of viral disease with applications in analytic and observational epidemiology diagnosis. Proteomic profiling allows deep insight into the pathogenesis of IHHNV molecular regulation and mechanism of hemocyte in Freshwater prawns.

Passiflora edulis is the major species of passionflowers grown worldwide, mainly for juice production and fresh fruit, in climates ranging from cool subtropical (purple variety) to warm tropical (yellow variety). The bacterial leaf spot, caused by Xanthomonas axonopodis pv. passiflorae (Xap) can be a serious disease affecting passion fruit production in commercial orchards, particularly under moist field conditions. In this study we describe a first analysis of host gene expression in this pathosystem. We used suppression subtractive hybridization to construct two cDNA libraries enriched for transcripts induced and repressed by Xap, respectively, 24h post inoculation with a highly virulent strain. High-quality sequences were obtained resulting in 998 unisequences that were used for annotation. In accordance with BLASTX results performed by Blast2GO tool, 86.7% of the unisequences showed similarity to other plant species proteins related to different functional categories. Sixty-three transcripts were similar to Arabidopsis thaliana defence-related proteins identified in the PLAZA platform. In silico predicted protein-protein interactions were detected on the basis of the STRING database for 35 of the 63 defence-related proteins. At this early stage of interaction, a set of genes was selected from Blast2GO categorization results and analyzed by quantitative PCR (qPCR). The expression profiles changed in response to the pathogen for 76% of these genes (48/63) and the differences in expression ratios ranged from 0.51-fold to 1.83-fold. In later stages of interactions (5 and 9 days post inoculation) when disease-associated symptoms were visible, qPCR analyses were performed for 14 genes selected from both libraries. The expression profiles of all genes were found to be changed by the pathogen. The gene that responded most strongly to the pathogen attack encodes a lipoxygenase 2. In inoculated plants, its expression was induced 500-fold and 300-fold, 5 and 9 dpi, respectively, compare!rn d to controls, suggesting an important role of this gene in passion fruit defence. Moreover, we showed that most of the genes involved in well-known pathogen recognition signaling pathways were repressed by Xap and this lends support to the idea that the jasmonic acid signaling pathway fails to be activated during the first hours of interaction.

Sri Janani J is currently pursuing her BTech Biotechnology in Karunya University, India. She has got Internship at Ben Gurion University, Israel from July 5th-31st 2015. She has participated in EVOGEN 2013-National Level Technical Symposium. She has also participated in Mindkraft 2K’13-National Level Techno Management Fest. She has been ranked 28th position in Coimbatore city at 10th National Science Olympiad.

Nanotechnology is the manipulation of any matter on atomic, molecular and supramolecular scale. Treating the anti-microbial activities against oral pathogens using nanotechnology is a well known techniquebut treating them with Zirconyl nitrate along with Curcuma longa has not been reported earlier. It is a unique combination where most scrutinized literature was collected after meticulous research. A comparative study of Zirconyl nitrate and zirconium oxychloride has been carried out by synthesizing them chemically by a certain process to know a better result. Zirconyl nitrate showed a better and also a clearer result in different parameters. So it has been chosen for further synthesis. A green synthesis has been carried out using Curcuma longa by extracting the longa. The extract was then treated with Zirconyl nitrate solution and later heated. After removing the impurities, the obtained product has been calcinated where a powder is procured. Subsequently the obtained powder is analyzed by Scanning Electron Microscope (SEM), X-Ray Diffraction(XRD), UltraViolet (UV) and Particle size analyzer methods where it confirmed the crystalline nature and morphology of the synthesized products. Streptococcus mutans and Escherichia coli has been selected to test the anti-microbial activities in the oral pathogens. It is selected to treat Oral thrush caused by fungus Candida albicans which accumulates on the lining of the human mouth. Zirconyl nitrate was studied on bacterial strains of E.coli and Streptococcus mutans. On the basis of the study, it could be speculated that the Zr (NO3)4 nanoparticles with the different active facets will show different antimicrobial activity.

HarishT has completed BTech Biotechnology from Karunya University 2013 and currently pursuing final year MTech Biotechnology from Karunya University, India. He had presented a review paper of “An Overview of Antibiotic Resistance Microorganisms” in the “International Symposium on Innovations in Free Radical Research and Experimental Therapeutics and 5th Annual Convention of Association of Biotech and Pharmacy” (2011) at Karunya University. He also had presented a poster under the title of “Nanotechnology in Clinical Diagnosis” in the “National Conference on “Molecular Diagnostics for Sustainable Health in Biosciences and Technology “held at Karunya University (2011). He has also participated in “Stimulating Bio-Entrepreneurs Talk” conducted by ABLE Biotech. He also attended national level workshop on “Recent Techniques in Cell Culture Cytotoxity Assay” at Karunya University and 3 National levels Technical symposium and an “International Symposium on Translational Neuroscience and 32nd Annual Conference of Indian Academy of Neurosciences” at NIMHANS, Bangalore, 2014.

A study was undertaken to isolate B. thuringiensis from sugarcane ecosystem in Tamil Nadu, India and to identify isolates containing crystalline protein of Bt that are toxic to Coleopteran insects. The isolation of B. thuringiensis was carried out on Traver’s media following heat treatment. Isolates were identified based on the ability of the isolate to produce crystal toxin which can be detected under phase contrast microscope. The cry gene content of 58 B. thuringiensis isolated from this study was identified by Polymerase Chain Reaction Method. The universal primers of cry 8 gene which is specific against the members of scarabaeidae family of the order Coleoptera was used to detect environmental isolates that would be positive for the gene. Isolate Bt 378 which was found cry8 positive revealed that the amplified nucleotide sequence when analyzed with ClustalW2 showed a maximum identity of 88.92% with other cry 8 genes. Further confirmation on its toxicity is being confirmed on the coleopteran beetle Holotrichia serrata (Fabricius).

Monika Bansal completed her PhD on the topic “Genetic transformation of Tomato for introduction of salinity tolerance. She worked as Senior Research Fellow at Haryana Agriculture University (June 2003-Dec 2003). Later she completed her Post-Doctoral fellow in the UGC funded project “Isolation and characterization of stress Inducible Promoter from Tomato\" at Delhi University South Campus with Dr. Arun Sharma (Aug 2008 –Aug2011). She joined as Assistant Professor at Asian Institution Affiliated to Punjab University, Patiala during August 2011-July 2013). From August 2014 till date she is working as Assistant Professor at School of Agriculture, Lovely Professional University.

Environmental stress such as drought, salinity and extreme temperatures are the major factors limiting plant growth and productivity. Cold stress leads to diverse change in transcription and translation of genes involved in developmental and physiological metabolism of plants. Constitutive over expression of transgenes imparting stress tolerance may hamper plant growth and productivity. It is desirable to produce transgenic plants that accumulate transgenic products only under stress conditions. To identify stress related transcripts we had utilized cold subtracted cDNA library to monitor gene expression changes after 24 hours of cold stress in tomato seedlings. Relative mRNA levels of cold stress associated gene CAT01 (NAC type transcription factor) from cold treated sample was compared with wild types. Sequence upstream of start codon from this transcript was selected to design the primers, promoter sequence was fused with GUS reporter gene and the recombinant transgenes was introduced into tomato cv. Pusa Ruby with Agrobacterium tumefaciens mediated genetic transformation. Histo-chemical and florometric analyses revealed promoter is inducible under all abioticstress conditions. This promoter is effective and desirable for deriving a low constitutive transgene expression under normal condition and high induction in response to cold, salt and drought stress. This method shall be advantageous as it will not have a major impact on plant biomass and yield.