Molecules and Methods for the Control of Biotic Stress Especially the Insect Pests — Present Scenario and Future Perspective

Continuous increase in population has unbalanced the demand and supply of agricultural produce. In this scenario, food security in a sustainable manner is being challenged due to several factors. Insect pests are considered as one of the major factors, which accounts for 35–100% crop damage, worldwide. Synthetic insecticides contributed significantly, but several safety concerns are associated with them. Transgenic crops with enhanced biotic or abiotic stress tolerance have shown promising contribution in achieving greater crop productivity. Transgenic cotton expressing Cry toxin of Bacillus thuringiensis has tremendously increased the production as well as the societal status of farmers in our country. However, a concomitant increase in the population of minor pests like aphids, whiteflies and others has demanded certain new approaches. Researchers have isolated several other toxic proteins like lectins, protease inhibitors, amylase inhibitors, chitinases, and tried various novel approaches like gene pyramiding, tissue specific expression and modulation in metabolites expression to combat emerging problems of insect pests. Conversely, the emergence of a new type of crop insect pests demands more specific effort for each insect. Besides this, there are several safety and ethical concerns that are associated with the use of genetically modified crops, which also need to be resolved as per demand. Development of a dedicated scientific forum for the proper demonstration of advantages and disadvantages of genetically modified crops to the citizens at ground level might be useful in resolving the societal and ethical concerns in our country

Role of Next-Generation RNA-Seq Data in Discovery and Characterization of Long Non-Coding RNA in Plants

The next-generation sequencing (NGS) technologies embrace advance sequencing technologies that can generate high-throughput RNA-seq data to delve into all the possible aspects of the transcriptome. It involves short-read sequencing approaches like 454, illumina, SOLiD and Ion Torrent, and more advance single-molecule long-read sequencing approaches including PacBio and nano-pore sequencing. Together with the help of computational approaches, these technologies are revealing the necessity of complex non-coding part of the genome, once dubbed as “junk DNA.” The ease in availability of high-throughput RNA-seq data has allowed the genome-wide identification of long non-coding RNA (lncRNA). The high-confidence lncRNAs can be filtered from the set of whole RNA-seq data using the computational pipeline. These can be categorized into intergenic, intronic, sense, antisense, and bidirectional lncRNAs with respect to their genomic localization. The transcription of lncRNAs in plants is carried out by plant-specific RNA polymerase IV and V in addition to RNA polymerase II and target the epigenetic regulation through RNA-directed DNA methylation (RdDM). lncRNAs regulate the gene expression through a variety of mechanism including target mimicry, histone modification, chromosome looping, etc. The differential expression pattern of lncRNA during developmental processes and different stress responses indicated their diverse role in plants

RNAi — Implications in Entomological Research and Pest Control

RNA interference (RNAi) has progressed swiftly in the past decade to become a convenient and dominant genetic tool that has immense utility in diverse fields. The entomological research, ranging from functional genomics to agriculture, has gained enormous momentum due to this technology. RNAi tool helped to discover the functions of new genes and study the complicated genetic networks, thus providing an evolutionary insight into various processes. RNAi is also becoming a method of choice for controlling insect pest populations. It is envisaged as tailor-made insecticide, which is highly species specific. However, the efficiency of this mechanism is limited by various factors such as the stability of the trigger molecule, the candidate gene selection, delivery system adopted and, most importantly, the choice of the target species. Apart from the successful implication in diverse areas, there are certain drawbacks of this technology such as ‘off-target’ effects, lack of sensitivity of various species, etc. Further research would relieve these limitations and support the manifestation of this genetic tool with much more reliability

Primary pulmonary non hodgkin’s T cell lymphoma

A 55-year-old female presented with history of fever, dry cough and right sided chest pain of two months duration. Radiological examination revealed a soft tissue attenuated lesion involving right middle lobe. Immuno histochemical analysis leads to the diagnosis of Non Hodgkin’s T-cell Lymphoma. The rarity of this disorder and its good clinical and radiological response to chemotherapy prompted us to report this case

Histopathological spectrum of neoplastic lesions of female reproductive system-a two-year experience in Eastern Nepal

Background: Neoplastic lesions of female reproductive system (FRS) consist majority of surgical cases. The aim of this study is to find out the frequency distribution of tumors occurring in FRS.Methods: This is a retrospective study carried out in Birat Medical College, situated in eastern Nepal. All formalin fixed surgical specimen of FRS from August 2015 to September 2017 were subjected for histopathology and examined under light microscopy. All neoplastic lesions were included, and non-neoplastic lesions were excluded from the study.Results: A total number of ninety (90) histopathological tissues representating various types of neoplastic lesion of FRS were studied. Out of which 6 (6.6%) tumors were from uterine cervix, (including 1 cervical leiomyoma and 5 squamous cell carcinoma cervix) 45(50%) tumors were from uterine body, (all leiomyoma) 20 (22.2%) tumors were from ovary (including 9 mature teratoma, 1 immature teratoma, 6 mucinous cystadenoma and 4 cases of serous cystadenoma) and 19 (21.2%) tumors were from breast (including 16 fibroadenoma, 1 lactational adenoma, 2 infiltrating ductal carcinoma and 1 comedo carcinoma). Majority 81(90%) were benign and 9 (10%) were malignant. Most common benign tumor was leiomyoma of uterine body and most common malignant tumor was squamous cell carcinoma of uterine cervix both commonly occurring in the age group of 41-50 years.Conclusions: Benign tumor, leiomyoma is the common tumor occurring in FRS. Carcinoma of the cervix is the common malignant tumor

Swyer- James -MacLeod syndrome presenting as hemoptysis in an adult

Swyer-James/MacLeod syndrome is an uncommon disease with characteristic radiological feature of unilateral hyperlucency due to loss of pulmonary vasculature and air trapping. Typically, this disorder is diagnosed in childhood during evaluations for recurrent respiratory infections. Here, we report a case in a 30-year-old adult female who presented with dyspnoea, cough with expectoration and recurrent hemoptysis due to associated bronchiectasis. This case highlights the importance of computed tomography in the diagnostic workup of recurrent hemoptysis in pulmonary tuberculosis epidemic countries like India

Genomic Dissection and Expression Profiling Revealed Functional Divergence in Triticum aestivum Leucine Rich Repeat Receptor Like Kinases (TaLRRKs)

The leucine rich repeat receptor like kinases (LRRK) constitute the largest subfamily of receptor like kinases (RLK), which play critical roles in plant development and stress responses. Herein, we identified 531 TaLRRK genes in Triticum aestivum (bread wheat), which were distributed throughout the A, B, and D sub-genomes and chromosomes. These were clustered into 233 homologous groups, which were mostly located on either homeologous chromosomes from various sub-genomes or in proximity on the same chromosome. A total of 255 paralogous genes were predicted which depicted the role of duplication events in expansion of this gene family. Majority of TaLRRKs consisted of trans-membrane region and localized on plasma-membrane. The TaLRRKs were further categorized into eight phylogenetic groups with numerous subgroups on the basis of sequence homology. The gene and protein structure in terms of exon/intron ratio, domains and motifs organization were found to be variably conserved across the different phylogenetic groups/subgroups, which indicated a potential divergence and neofunctionalization during evolution. High-throughput transcriptome data and quantitative real time PCR analyses in various developmental stages, and biotic and abiotic (heat, drought and salt) stresses provided insight into modus operandi of TaLRRKs during these conditions. Distinct expression of majority of stress responsive TaLRRKs homologous genes suggested their specified role in a particular condition. These results provided a comprehensive analysis of various characteristic features including functional divergence, which may provide the way for future functional characterization of this important gene family in bread wheat

Streamlined and Cost-Effective Genomic DNA Extraction Method for Lichens, Mushrooms, and Endolichenic Fungi: Enabling DNA Barcoding and Molecular Research

Extraction of nucleic acids in pure form from organisms is of paramount importance for DNA based identification and other molecular studies. Over the past few decades, DNA-barcoding has emerged as a powerful technique, facilitating species identification across various ‘difficult to identify’ life-forms. Fungi, being an immensely diverse group of microorganisms, contribute significantly to global biodiversity, with estimates ranging from 2.2 to 3.8 million species. However, a vast majority of this diversity remains unidentified, and many fungal species are considered cryptic. Therefore, numerous large- and small-scale DNA-barcoding projects are being conducted worldwide to unravel this rich biodiversity. However, the rigidity and high complex polysaccharides content of fungal cell-wall presents a significant obstacle, making the extraction of high-quality genomic DNA a challenging task across varied fungal organisms. In this study, we employed a modified CTAB based method to isolate and purify high-quality PCR-amplifiable genomic DNA primarily from lichens and tested it on other fungal life forms as well, including, mushrooms, endolichenic fungi, and parasitic fungi. Remarkably, the isolated DNA proved successful as a template in PCR reactions, serving the purposes of DNA barcoding, RAPD as well as for metagenomic analysis effectively. This versatile protocol demonstrated its utility across all the fungal life forms investigated in this study, offering a universal, cost-effective, and efficient approach for fungal DNA isolation

NMR and molecular modelling studies on the interaction of fluconazole with β-cyclodextrin

<p>Abstract</p> <p>Background</p> <p>Fluconazole (FLZ) is a synthetic, bistriazole antifungal agent, effective in treating superficial and systemic infections caused by <it>Candida </it>species. Major challenges in formulating this drug for clinical applications include solubility enhancement and improving stability in biological systems. Cyclodextrins (CDs) are chiral, truncated cone shaped macrocyles, and can easily encapsulate fluconazole inside their hydrophobic cavity. NMR spectroscopy has been recognized as an important tool for the interaction study of cyclodextrin and pharmaceutical compounds in solution state.</p> <p>Results</p> <p>Inclusion complex of fluconazole with β-cyclodextrins (β-CD) were investigated by applying NMR and molecular modelling methods. The 1:1 stoichiometry of FLZ:β-CD complex was determined by continuous variation (Job's plot) method and the overall association constant was determined by using Scott's method. The association constant was determined to be 68.7 M^-1which is consistent with efficient FLZ:β-CD complexation. The shielding of cavity protons of β-CD and deshielding of aromatic protons of FLZ in various¹H-NMR experiments show complexation between β-CD and FLZ. Based on spectral data obtained from 2D ROESY, a reasonable geometry for the complex could be proposed implicating the insertion of the <it>m</it>-difluorophenyl ring of FLZ into the wide end of the torus cavity of β-CD. Molecular modelling studies were conducted to further interpret the NMR data. Indeed the best docked complex in terms of binding free energy supports the model proposed from NMR experiments and the <it>m</it>-difluorophenyl ring of FLZ is observed to enter into the torus cavity of β-CD from the wider end.</p> <p>Conclusion</p> <p>Various NMR spectroscopic studies of FLZ in the presence of β-CD in D₂O at room temperature confirmed the formation of a 1:1 (FLZ:β-CD) inclusion complex in which <it>m</it>-difluorophenyl ring acts as guest. The induced shift changes as well as splitting of most of the signals of FLZ in the presence of β-CD suggest some chiral differentiation of guest by β-CD.</p