Background: SARS-CoV-2 Non-Structural Protein 6 (NSP6) is pivotal for viral replication, but a comprehensive understanding of its evolutionary stability, functional sites and dynamic behavior has been limited. Leveraging a newly established high-confidence 3D model, this study provides an integrative analysis of NSP6&rsquo;s biology and biophysics. 
 
Methods: The validated NSP6 structure was analyzed for conserved regions and evolutionary history using BioEdit and MEGA11 for phylogenetic tree construction. Functional motifs and post-translational modification (PTM) sites were predicted using PROSITE, SMART, MotifFinder and MotifScan. Structural classification was performed using CATH, SCOP and SUPERFAMILY. A 100 ns molecular dynamics (MD) simulation was conducted using GROMACS with the CHARMM27 force field to evaluate structural stability through RMSD, RMSF, Rg and SASA. 
 
Result: Phylogenetic analysis revealed NSP6&rsquo;s close relationship to bat coronaviruses and identified a single, fully conserved domain across its entire 290-amino-acid length. Motif analysis identified the definitive Coronavirus replicase NSP6 domain and predicted critical PTM sites, including Casein Kinase II phosphorylation and N-myristoylation sites. Structural classification revealed an unexpected homology to cobalamin adenosyltransferase-like folds. The 100 ns MD simulation demonstrated outstanding model stability, with low RMSD (0.2-0.35 nm after 20 ns), a consistent radius of gyration (2.04&plusmn;0.01 nm) and stable solvent-accessible surface area.

Evolutionary Conservation, Functional Motifs and Molecular Dynamics of SARS-CoV-2 NSP6

This review analyses the use of molecular techniques in rumen microbial identification. The use of regular methods like roll tube and most probable number resulted in under estimation of rumen microbial growth. So molecular biology acts as an advanced tool for rumen microbial culture and identification of various new species. These techniques give complete descriptions of individual ruminal populations. Use of molecular techniques like PCR, DDGE and FISH, which also pave pathway for genetic manipulation of rumen microbes in the field of rumen manipulation. So the combination of traditional and molecular assays gives accurate and satisfactory results.

Molecular techniques in rumen biotechnology: A review

Molecular markers are effective tools used to &lsquo;flag&rsquo; the location of a specific gene or the inheritance of a definite trait. Markers are unique DNA fragments that can be identified inside the entire genome. The development of molecular markers combined with high throughput technologies have paved the way for achieving the desirable traits as well as induced biotic and abiotic stress tolerance in plant, which enhanced the crop breeding. Highly polymorphic molecular markers are developed for gene mapping, estimation of genetic diversity, finding out the evolution and phylogeny of crop, analysis of heterosis, assessment of diploid/haploid crops and genotyping of cultivars along with Marker Assisted Breeding (MAB)/Marker Assisted Selection (MAS). These are the most significant objectives for crop breeding. This review reveals about the role of various recently developed molecular markers in the improvement of crop. Molecular markers act as a &ldquo;milestone&rdquo; for the researchers who aim to enhance crop breeding.

Role of Molecular Markers in Crop Breeding: A Review

<div style="text-align:justify">An investigation was carried out to study the antioxidant, anticancer and docking activities of medicinal plant, Tulsi &ndash; Ocimum sanctum. The results of antioxidant activity of Tulsi leaves extract showed that tulsi leaves has good free radical scavenging ability and IC50 value was found to be 54.23% at 500 &micro;g/ml. The results of anticancer activity of Ocimum santum on normal VERO cell and MCF-7 cell line has showed good anticancer activity having the IC50 concentration at 51.1 &igrave;|g/ml against MCF-7 cell lines &ndash; Human Breast cancer cell line thereby indicating that the percentage of cell viability increases with the increased concentration of Ocimum santum (Tulsi) whereas 24.4% cell viability was recorded in the normal VERO cell line. DNA laddering assay was performed on agarose gel electrophoresis. A clear fragmented DNA ladders were observed in tulsi treated MCF-7&ndash; Human Breast cancer cell lines. &nbsp;But the untreated normal VERO cell lines did not show any DNA fragmentation. Thereby confirms that Ocimum sanctum induced cell death on breast cancer cell line MCF-7 cancerous cell through apoptosis. Molecular docking was carried out (AutoDock tools), Docking simulations was performed using Lamarckian genetic algorithm of Solis and Wests local search method. Interaction between HDAC6 and Cirsilileon possessing binding energy showed +15.91kcal for interaction studies. It was found that most of the hydrogen bonding is with the residues TYR 76, TRP 35, SER 67, TYR 81, GLU 33, TYR 48, ILE 69 and MET 53 present in the binding pocket. The ligand was docked with the target protein, and the best docking poses were identified and the binding poses of the cirsilieon was shown. This act as the best docking poses shows how the ligand molecule fits into the binding region of the target protein.&nbsp;</div>

Antioxidant, anticancer and molecular docking activity of tulsi plant

<div style="text-align:justify">Rapid industrialization and successful green revolution have introduced a wide array of chemicals into our environment; some of these chemicals entered in ecosystem; gets accumulated and exert serious health and ecological problems. These toxic substances can enter the food chain and emphasize pathological changes which damage either cell, organ or system (circulatory, immune, respiratory, digestive, nervous, reproductive and musculo-skeletal etc.) by altering structure and/or function of biological components; DNA, RNA, proteins, lipids and carbohydrates. This review article provides certain molecular diagnostic techniques used for their robust and accurate detection at molecular level. Investigations conducted during 2020-2021 where various review and research articles were surveyed and then extracted to enlist congestive datum for rapid detection of toxicological changes in animals. In our investigations we concluded that toxic substances present in our environment affects health of animals by altering structure and functioning of biomolecules and their concerned system. These cytological and systemic changes can be detected with the help of molecular diagnostic techniques including dideoxysequencing, pyrosequencing, allele specific RT-PCR, CRISPER/Cas, genotyping and microarrays etc. present collection of data will provide congestive information for rapid toxicological detection at molecular level.</div>

Molecular Diagnostic Strategies for the Detection of Toxicological Changes in Animals: A Review

Background: Groundnut (Arachis hypogaea L.) construed as one of the most dynamic oilseed crop, delivering high protein besides significantly meet country&rsquo;s major oil requirement. In India, 10.1 million tonnes of groundnut are produced over an area of around 5.7 million hectares at a productivity of 1777 kg/ha. Groundnut is always been a fascinating crop to small and marginal farmers but its yield was greatly challenged by foliar fungal diseases such as early leaf spot, late leaf spot and rust. Among these, late leaf spot (LLS) is a major foliar fungal disease which cause severe defoliation of diseased leaflets and reduce pod and fodder yield by 50 per cent besides the kernel quality. 
 
Methods: The present investigation was conducted at Agricultural College and Research Institute, Killikulam during the year 2022-2024 involving 220 groundnut genotypes along with a resistant check (ICG 6066) and susceptible checks (TMV 2, TMV 7 and TMV 13) for screening against late leaf spot resistance under both field and artificial controlled conditions thereby assessment of the disease through molecular intervention.&nbsp; 
 
Result: Molecular interpretation revealed five markers exhibiting polymorphism to LLS disease out of 30 markers validated. The PIC values ranged from 0.501 (TC7H11) to 0.570 (Ah3TC23H10). Molecular marker screening for disease resistance categorized ICG 978, ICG 1649, ICG 1703 and ICG 2280 genotypes as resistant to late leaf spot disease&nbsp; while ICG 8760, ICG 4598, ICG 5051, ICG 5891 and ICG 6057 exhibited moderately resistant for late leaf spot besides exhibited higher mean yield performance among the test genotypes. Hence, the genotypes ICG 8760, ICG 4598, ICG 5051, ICG 5891 and ICG 6057 may be utilized for developing ideal recombinants for LLS resistance breeding in Groundnut.

Molecular Assessment of Late Leaf Spot Resistance in Germplasm Collections of Groundnut (Arachis hypogaea L.)

Ca-B interactions in crop plants are essential in crop productivity, quality and stress-tolerance in a variety of agro-climatic conditions. This review discusses the complex processes behind Ca-B nutrition in plants, both at the molecular interaction level and in the field. Calcium ionically bridges with the pectin compounds at the cellular level and boron forms borate crosslinking with rhamnogalacturonan II (RG-II). The resulting B-RG-II complex that involves two molecules of boric acid and two Ca2+&nbsp;ions is important to retain the cell wall integrity. The effect of these nutrients during the plant development is synergistic and when there is deficiency of boron, it leads to increased cytosolic Ca2+&nbsp;and also calcium-regulated gene expression. Physicochemical properties of soil play a great role in Ca-B dynamics. Boron adsorption on clay minerals is enhanced as pH rises reaching a peak of about pH 9.0 in alkaline environment. The arable land of the world is estimated at around 50% of the global land, with 50% of this area being acid in nature making the availability of the two nutrients to be limited and thus requires strategic interventions. Optimised Ca-B nutrition leads to significant increases in yield of major crops. This can only be achieved through combined methods of soil testing, precise fertilisation and environmental concerns to facilitate good management. Nutrient management strategies are being transformed with the new technologies such as genomics, metabolomics, precision agriculture, mycorrhizal associations and machine learning algorithms. New areas of research should be directed towards new decision support systems, molecular level interventions, sustainable boron recovery of waste streams and mycorrhizal-mediated nutrient interactions. 
&nbsp;

Calcium-boron Interactions in Plant Nutrition: From Molecular Mechanisms to Agricultural Applications: A Review

<div>Background: Pathomicrobial and molecular investigations of respiratory system diseases was undertaken on twelve adult buffalo carcasses received for necropsy examination at post mortem facility of Department of Veterinary Pathology, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana during the period from August, 2018 to February, 2019.&nbsp; 
 
Methods: After collection of the samples, laboratory work was undertaken in the laboratories of Department of Veterinary Pathology, Department of Animal Biotechnology and Central Laboratory of the College, LUVAS, Hisar, Haryana (India) in the year 2018-2019 regarding the examination of the clinical history, pathological, microbiological and molecular investigations. 
 
Result: The gross pathological changes observed in lungs were variable degree of vascular changes, mild to severe consolidation, fibrin deposition along with adhesions to the thoracic wall. Histopathological findings revealed abnormalities of inflation such as pulmonary emphysema, atelectasis, pulmonary congestion and haemorrhages which was associated with different types of pneumonia viz. fibrinous bronchopneumonia, suppurative giant cell pneumonia, interstitial pneumonia and serous pneumonia. Lung, heart blood and tracheal swab samples collected from the buffalo carcasses revealed eighteen bacterial isolates which were identified by Vitek 2 system. These include E. coli (11 isolates), Salmonella enteric enterica (2 isolates), Acinetobacter ursingii (1 isolate), Staphylococcus haemolyticus (1 isolate), Staphylococcus sciuri (1 isolate), Staphylococcus warneri (1 isolate) and Staphylococcus hominis (1 isolate) mostly belonging to opportunistic pathogen category. E. coli serotypes confirmed from these cases were O83, O149 and O8. The results of in vitro drug sensitivity testing revealed that most of bacterial strains were found sensitive to cefoperazone/sulbactum and co-trimoxazole. Molecular studies confirmed Bovine Herpes Virus-1 (BHV-1) infection in one case with Fibrinous bronchopneumonia through real-time quantitative PCR indicating the prevalence of the infection in the state.</div>

Pathomicrobial and Molecular Investigations of Respiratory System Diseases Affecting Buffaloes

<div>Background: Riemerella anatipestifer (R. anatipestifer) is a gram negative, microaerophilic, non-motile, bipolar bacteria. High genetic diversity and molecular differentiation were reported among field isolates. Although the bacterium causes one of the most economically important duck diseases in the north-eastern region of India, little work has been done on isolation, identification and molecular characterization of the bacteria. Hence, the present investigation was undertaken with a view to characterize the R. anatipestifer isolates from ducks of Assam.</div>

<div> 
Methods: Phenotypic and molecular identification of R. anatipestifer isolates from domesticated ducks of Assam, India were carried out during the period from February, 2019 to January 2020. A total of 624 samples (Ocular swab, throat swab, liver, spleen, kidney, brain, heart, lung) from ducks comprising of apparently healthy, ailing and dead ducks were collected from five districts of Assam, India were processed to isolate and identify the bacteria. The tentative identification of the bacteria was done based on phenotypic characteristics viz., colony morphology, growth characteristics and biochemical reactions. All the phenotypically positive isolates were further subjected to molecular identification based on PCR assay targeting 16S rRNA gene and ERIC sequence. 
 
&nbsp;</div>

<div>Result: The bacteria could be isolated from different field samples. The highest percentage of the samples that yielded the bacteria are from brain (76%) followed by spleen (74%) of dead ducks and less number of ocular swab (33%) from apparently healthy ducks were found positive. Sequencing of the amplified product of the selected R. anatipestifer isolates targeting 16S rRNA gene revealed homology percentage of 96.5-100%. Further, sequences representing five geographical locations were submitted to NCBI gene bank. Phylogenetic studies of the isolates indicated that there is prevalence of at least two genetically different strains of R. anatipestifer in the study area. The study suggested that the R. anatipestifer infection is endemic in Assam causing varying rate of morbidity (39%) and mortality (53%) and molecular based confirmation is necessary besides phenotypic identification.</div>

<div>Isolation and Molecular Characterization of Riemerella anatipestifer from Domesticated Ducks of Assam, India</div>

<div>Background: The estimation of genetic diversity in pigeonpea is crucial for desiging breeding programmes and germplasm conservation. Morphological studies alone do not provide sufficient information to understand genetic diversity. Molecular analysis using SSRs can provide additional information on genetic diversity that can be used for selection of diverse parents. 
 
Methods: The experimental material for present study consisted of 50 diverse genotypes of pigeonpea. The fifty genotypes were planted in Randomized Complete Block Design consisted of three replications during the kharif 2020-21. The estimation of morphological genetic diversity was done by Mahalanobis D2 statistics. The molecular genetic diversity analysis was done by using 30 molecular markers in same genotypes. 
 
Result: The assessment of morphological diversity revealed that the fifty pigeonpea genotypes were grouped into four different clusters with cluster I as the largest cluster (33 genotypes). The molecular markers differentiated the 50 genotypes in five major clusters with cluster II as the largest cluster (24 genotypes). The results of present study suggested that morphological and molecular diversity in pigeonpea is different. On the basis of genetic distance, the genotypes RVSA 2014-1 and PA 406 were found most genetically distant and may be used in hybridization programme to create diverse progenies.</div>