Single stranded DNA viruses (Genus Begomovirus) are economically important plant pathogens that are responsible for significant yield losses in dicotyledonous plants including radish (Raphanus sativus L.). However, their distribution and diversity in certain regions of Pakistan are still not sufficiently described. During systematic field surveys conducted across six districts of Punjab province (Faisalabad, Okara, Toba Tek Singh, Multan, Lahore, and Layyah), more than 120 radish fields as well as vegetable markets were inspected, but the infected plants were detected only in six localities of Thal desert in district Layyah. The radish plants exhibiting vein thickening, leaf shortening, and enations were observed, suggestive of a geminivirus infection. Total DNA isolated from symptomatic leaves was analyzed by diagnostic PCR. Amplification using coat protein primers targeting the DNA-A component yielded a 771 bp fragment (GenBank Accession No. PV872843), while universal primers for betasatellites amplified a 1361 bp fragment (GenBank Accession No. PV872842). The sequence similarity analysis revealed that DNA-A component shared similarity with Pedilanthus leaf curl virus, whereas the associated betasatellite showed 85% nucleotides identity with radish-infecting betasatellites. According to the betasatellite species criteria established by ICTV which defines distinct species at less than 91% nucleotide sequence identity, this isolate represents a divergent species for which the name Radish leaf curl Layyah betasatellite is proposed. The phylogenetic tree for coat protein suggested its close clustering with Pedilanthus leaf curl virus and Cotton leaf curl Kokhran virus reported from the region. However, betasatellite clustered with papaya infecting betasatellite previously reported from radish crops in India. This study documents, the natural occurrence of Pedilanthus leaf curl virus and a novel Radish leaf curl Layyah betasatellite infecting radish in Pakistan. Our findings underscore the occurrence of begomovirus with new betasatellite complexes in the region and highlight the need for continuous surveillance strategies to safeguard radish cultivation.

Silybum marianum (SM) is a rich source of flavonolignans with promising antioxidant and antidiabetic properties; however, its therapeutic application is limited by poor stability and bioavailability. This study combined experimental and computational approaches to develop and evaluate SM-loaded chitosan nanoparticles (CS-SM nanoparticles). Microwave-assisted extraction followed by LC-MS/MS profiling identified eleven metabolites, including major flavonolignans characteristic of SM. Nanoparticles prepared by ionic gelation exhibited favorable physicochemical properties, including a particle size of 173-189 nm, a polydispersity index of 0.23, a zeta potential of +41.5 mV, an encapsulation efficiency of 98%, and a drug loading capacity of 50%, indicating the formation of a stable colloidal delivery system. CS-SM nanoparticles showed enhanced antioxidant, anti-inflammatory, and α-amylase inhibitory activities compared with crude extracts. The formulation exhibited an α-amylase IC50 value of approximately 0.40 mg/mL and maintained low hemolytic activity, suggesting favorable preliminary biocompatibility. Molecular docking demonstrated favorable interactions of neosilyhermin A, silibinin, and silyhermin with α-amylase and α-glucosidase active sites. Short-timescale molecular dynamics simulations revealed ligand-dependent behavior within the chitosan-TPP matrix, indicating different release tendencies among the investigated flavonolignans. Overall, the results support CS-SM nanoparticles as a promising platform for the delivery of bioactive phytochemicals with antioxidant and antidiabetic potential.

Cancer continues to be a major global health burden, with receptor tyrosine kinases such as EGFR, ERBB2, and VEGFR-3 being critical therapeutic targets due to their central roles in tumor growth, survival, and angiogenesis. Current therapies, while effective in some contexts, face limitations including resistance, toxicity, and high cost, highlighting the need for novel multi-target approaches. In this study, we report the isolation and computational characterization of a novel defensin-like peptide (DEFL) from Datura stramonium (GenBank accession KT371458). The peptide sequence encoded 74 amino acids and displayed characteristic cysteine-stabilized motifs. Docking simulations revealed favorable binding scores toward EGFR (- 80.6 ± 10.6), ERBB2 (- 63.6 ± 7.4), and VEGFR-3 (- 50.5 ± 6.7), with interactions involving residues located within predicted receptor-binding regions. To further assess stability, 100 ns molecular dynamics simulations were performed. RMSD profiles confirmed stable complexes, with EGFR stabilizing around 0.6-0.8 nm, ERBB2 around 0.7-0.9 nm, and VEGFR-3 at a tighter 0.3-0.4 nm. Ligand RMSDs indicated moderate flexibility for ERBB2 (peaks up to 1.3 nm) but tighter stability for VEGFR-3 (0.3-0.5 nm). RMSF analyses revealed minimal fluctuations (< 0.3 nm) at binding sites, and radius of gyration values remained stable, indicating compact receptor-peptide complexes (EGFR: 3.45-3.75 nm; ERBB2: 2.95-3.20 nm; VEGFR-3: 1.92-1.98 nm). Hydrogen bond profiling and additional trajectory analyses (DCCM and PCA) supported overall system equilibration without major structural disruption during the simulations. The Datura stramonium defensin-like peptide indicating a stable and energetically favorable peptide-receptor interactions at the computational level. Overall, the simulations indicate persistent peptide-receptor association and stable structural behavior of the complexes at the computational level. However, molecular docking and molecular dynamics simulations do not demonstrate functional inhibition of EGFR, ERBB2, or VEGFR-3, nor do they confirm anticancer efficacy. Therefore, these results should be interpreted strictly as hypothesis-generating in silico predictions, and experimental validation, including peptide synthesis, receptor-binding assays, extracellular-domain competition assays, and cancer cell-based functional studies, will be required to confirm biological relevance.

Antimicrobial resistance (AMR), driven largely by ESKAPE(E) pathogens, represents a critical global health challenge. The increasing prevalence of multidrug-resistant (MDR) bacteria and the declining effectiveness of conventional antibiotics have created an urgent need for innovative therapeutic agents with novel mechanisms of action. In this study, we aimed to establish a stability-centric computational discovery pipeline to identify and characterize novel antimicrobial peptides (AMPs) with potent and selective bactericidal activity against ESKAPE pathogens, and to validate the lead candidates through experimental in vitro assays. A library of approximately 3,000 AMPs was retrieved from public databases and subjected to in silico screening for physicochemical properties and predicted toxicity. Following filtration, 200 peptides were selected as ligands for molecular docking against essential target proteins from ESKAPE pathogens. Based on docking scores, binding pocket occupancy, and ligand-protein interaction, the top 10 peptides against each bacterium were shortlisted. The four top consensus hits of Vespid Chemotactic Peptide VT1 (VCP-VT1), Taromycin A, CN-AMP1, and Alliumin were selected based on their superior inhibitory profiles. Subsequent peptide-protein docking analyses confirmed their binding modes and interaction patterns. These four candidates were advanced to peptide-protein interaction profiling, followed by pilot in vitro minimum bactericidal concentration (MBC) determination and cytotoxicity assessment on a mammalian cell line. Hierarchical consensus scoring identified VCP-VT1, Taromycin A, CN-AMP1, and Alliumin as the top-ranked binding AMPs. VCP-VT1 was the most potent peptide in preliminary MBC assays, with modest activity against Gram-positive Enterococcus faecalis, Enterococcus faecium and Staphylococcus aureus, and Gram-negative Acinetobacter baumannii and Pseudomonas aeruginosa (MBC50 of 50 µM, MBC > 90 of 100 µM). All peptides showed minimal cytotoxicity against a cultured human cell line and non-toxic ADMET parameters in silico. This reductionist validation of a stability-pre-filtered computational shortlist substantiated VCP-VT1 as a priority bactericidal lead AMP. Its favourable preliminary efficacy and safety margins would warrant further pharmacokinetic optimization and eventual in vivo efficacy studies against recalcitrant multidrug-resistant (MDR) pathogens. This integrated computational-experimental framework provided a generally applicable strategy for accelerating the discovery of peptide-based therapeutics to combat the growing threat of AMR.

The dramatic surge of neurodegenerative disorders among elderly population underscore the pressing demand for development of optimal and evidence based noninvasive natural treatment strategies. Paraquat exposure in animal models used in scientific studies can cause a variety of clinical signs of Parkinson disease (PD). The health benefits of thyme include antioxidant, anti-inflammatory, pulmonary, and neurological benefits. Thyme and other herbal treatments are frequently used to treat a variety of conditions, including neurological issues. The primary factor in the etiology of neurodegeneration is oxidative stress. Conventional treatments are indicated to potentially have negative side effects. The primary phytochemicals of Thymus vulgaris (TV), which are responsible for its unique therapeutic property of neuro-protection, include hydrocarbon and phenolic compounds like thymol and carvacrol. The goal of the current investigation was to examine T. vulgaris' potential for neuroprotection while also ensuring its safety. Analyses of the plant's physicochemical and phytochemical composition were performed by liquid chromatographic analysis. Neuro-behavioral and biochemical parameters were evaluated to determine the impact of T. vulgaris in paraquat induced parkinsonian rodents model. The neurobehavioral tests include open field tests for movement and exploration, Y maze test and elevated plus maze test for natural behavior, memory, and anxiety, hole board tests for exploratory behavior, ladder climbing, foot printing, and wire hanging tests for estimating neuromuscular coordination. T. vulgaris treatment significantly improved neurobehavioral parameters dose-dependently, Biochemical analysis revealed that extract treatment mitigated the declined level of antioxidant enzymes. RT-PCR analysis showed that in paraquat treated group mRNA expression of IL-1α, IL-1β, Alpha-Synuclein, TNF-α, and IL-6 was upregulated markedly. However, T. vulgaris treatment dose dependently down-regulated the mRNA expression of these genes. The groundbreaking results of current study revealed that T. vulgaris restored the degenerative alterations, neuro-inflammation, and nerve loss in the brain structure, as evident by histopathological investigation. Particularly remarkable restoration in neuropsychological and biochemical markers emphasize the medicinal potential of T. vulgaris as a revolutionary treatment for neurodegenerative disorders, offering new hope for millions worldwide afflicted by these devastating conditions.

Cotton leaf curl Multan virus (CLCuMuV), is a begomovirus encoding a protein, Replication associated protein (Rep) infect cotton crop. If the function of this protein is disrupted virus fails to replicate and subsequently cannot complete its life cycle. In this study plant-based chemicals were tested as inhibitors of Rep protein of CLCuMuV. In this study, sequence and docking analysis, calculations of physicochemical parameters and three-dimensional structural modeling of Rep protein of CLCuMuV against selected phytochemicals were performed. Based on docking analysis the best phytochemicals were reported. Analysis of the sequence and physicochemical properties revealed the Rep protein as negatively charged, slightly acidic, and prone to instability. The three-dimensional model of the protein showed that it consists of two helices, nine beta-pleated sheets, and eleven coiled regions. More than 900 phytochemicals were used in the docking experiment. Ten phytochemicals were obtained as potential inhibitors of Rep protein based on docking score. Through the analysis of the interaction, the atoms that participate in each interaction between the Rep protein and phytochemicals were identified. The results indicated that phytochemicals are potential inhibitors of Rep protein of CLCuMuV. Our study will be useful for devising resistance strategies against CLCuMuV in cotton.

As the population of the world rises and the amount of farmable land decreases, the need for food grows ever more urgent. This calls for research into developing high-yielding cultivars that are both nutrient-dense and resistant to environmental challenges so that farmers can keep up with the demand. Mutagenesis, intergeneric crosses, and translocation breeding are only a few examples of the methods explored to enhance crop quality. As genetic engineering progressed, transgene insertion became a common method for improving the resilience of genetically engineered crops to environmental stresses. Genetically modified (GM) crops are regulated based on an assessment of the risks they pose to the environment and the public's health, using either a process- or a product-focused approach. Recent advances in gene-editing technology have opened the door to a new era of plant breeding by making it possible to change genes precisely without transferring genes from other plants. Zinc finger nucleases, site-directed nucleases (SDNs), and the clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (Cas 9) are all examples of these kinds of techniques (Cas9). But the question, of whether or not to regulate such methods and whether or not the public will accept them persists. In contrast to the United States, which exempts gene-edited plants from the restrictions that apply to genetically modified organisms (GMOs), Europe and New Zealand classify gene-edited plants as GMOs. Depending on the regulatory strategy used in each nation or region, the commercialization of these crops and products may or may not require clearance from the relevant regulatory organizations. Because of the lengthy and complex clearance process necessary to employ them, their immediate commercial worth would be reduced if they were classified as GMOs. Strong regulatory measures for emerging technologies are needed to speed up crop research and encourage people to accept these new kinds of agricultural production with confidence.

The phytic acid, tannins, and trypsin inhibitors reduces protein digestibility in pulses. The objective of this study was to standardize the thermal application to improved protein quality. The flours treated with roasting at 120oC for 30 min showed percent reduction in phytic acid was as 29.75-35.47%, 27.44-36.34% and 28.23-45.46% in selected pulses respectively. Autoclaving showed percent phytic acid reduction in the range of 18.16-21.51%, 17.42-23.58% and 15.74-31.61%. % reduction of trypsin inhibition activity was in the range of 30.18-88.15% for chickpea, 35.25-92.05% for mung bean and 32.18-95% for lentil. Similar trend was observed in autoclaving with less reduction i.e., 27.25-75.22% in chickpea 30.23-89.04% in mung bean and 52.58-92% in lentil. An increase in protein (21.35, 19.96 and 23.40%) was confirmed. The extended lysine (0.374mg/100g) was observed in composite flour as compared to wheat flour (0.246mg/100g). It is evident that pretreated pulses flour improved the amino acid profile of end product.

Ginger oleoresins being rich in coloring, flavoring and other phytochemical compounds have strong potential for industrial applications. In this study, 5 different oleoresins’ extraction methods are compared in terms of yield and phytochemical characterization of oleoresins. Petroleum ether exhibited the highest yield (5.51 ± 0.08

Proteases hydrolyze proteins and reduce them to smaller peptides or amino acids. Besides many biological processes, proteases play a crucial in different industrial applications. A 792 bp protease gene (nprB) from the thermophilic bacterium Streptomyces thermovulgaris was cloned and expressed in E. coli BL21 using pET 50b (+). Optimal recombinant protease expression was observed at 1 mM IPTG, 37 °C for 4 h. The resulting protease was observed in soluble form. The molecular mass estimated by SDS-PAGE and Western blot analysis of the protease (NprB) fused with His and Nus tag is ~70 KDa. The protease protein was purified by Ammonium sulfate precipitation and immobilized metal ion affinity chromatography. The optimum pH and temperature for protease activity using casein as substrate were 7.2 and 70 °C, respectively. The mature protease was active and retained 80% of its activity in a broad spectrum of pH 6–8 after 4 h of incubation. Also, the half-life of the protease at 70 °C was 4 h. EDTA (5 mM) completely inhibited the enzyme, proving the isolated protease was a metalloprotease. NprB activity was enhanced in the presence of Zn2+, Mn2+, Fe2+ and Ca2+, while Hg2+ and Ni2+ decreased its activity. Exposure to organic solvents did not affect the protease activity. The recombinant protease was stable in the presence of 10% organic solvents and surfactants. Further characterization showed that zinc-metalloprotease is promising for the detergent, laundry, leather, and pharmaceutical industries.

The rise in antibiotic resistance in pathogens has led to an exponential rise in infectious diseases. It has alarmed the need for new antibiotics with precise actions against pathogens and ability to target multi directionally to encounter disease causing agents. Cationic peptides are also known as natural antibiotics and play a fundamental role in recruiting and promoting agents of innate as well as adaptive immune systems. These are produced constitutively as well as under the influence of biotic and abiotic stresses. In the current study, Solanum lycopersicum (tomato) was induced to express cationic proteins/ peptides under salt stress. Proteins up to molecular mass of 10 kDa were isolated and fractionated by cation exchange chromatography followed by antibacterial assays. The results showed that tomato plants could tolerate salt stress of 200 mM, where 100 mM of stress provoked more antibacterial peptides/ proteins. Strong cation exchange fractionation separated enormous positively charged peptides which showed high percentage of antibacterial activity. The prolific antimicrobial peptides authenticate the tomato plant as a promising candidate for pharmaceuticals, nutraceuticals and drug discovery.

Begomoviruses are contagious and severely affect commercially important fiber and food crops. Cotton leaf curl Multan virus (CLCuMuV) is one of the most dominant specie of Begomovirus and a major constraint on cotton yield in Pakistan. Currently, the field of plant genome editing is being revolutionized by the CRISPR/Cas system applications such as base editing, prime editing and CRISPR based gene drives. CRISPR/Cas9 system has successfully been used against biotic and abiotic plant stresses with proof-of-concept studies in both model and crop plants. CRISPR/Cas12 and CRISPR/Cas13 have recently been applied in plant sciences for basic and applied research. In this study, we used a novel approach, multiplexed crRNA-based Cas12a toolbox to target the different ORFs of the CLCuMuV genome at multiple sites simultaneously. This method successfully eliminated the symptoms of CLCuMuV in Nicotiana benthamiana and Nicotiana tabacum. Three individual crRNAs were designed from the CLCuMuV genome, targeting the specific sites of four different ORFs (C1, V1 and overlapping region of C2 and C3). The Cas12a-based construct Cas12a-MV was designed through Golden Gate three-way cloning for precise editing of CLCuMuV genome. Cas12a-MV construct was confirmed through whole genome sequencing using the primers Ubi-intron-F1 and M13-R1. Transient assays were performed in 4 weeks old Nicotiana benthamiana plants, through the agroinfiltration method. Sanger sequencing indicated that the Cas12a-MV constructs made a considerable mutations at the target sites of the viral genome. In addition, TIDE analysis of Sanger sequencing results showed the editing efficiency of crRNA1 (21.7%), crRNA2 (24.9%) and crRNA3 (55.6%). Furthermore, the Cas12a-MV construct was stably transformed into Nicotiana tabacum through the leaf disc method to evaluate the potential of transgenic plants against CLCuMuV. For transgene analysis, the DNA of transgenic plants of Nicotiana tabacum was subjected to PCR to amplify Cas12a genes with specific primers. Infectious clones were agro-inoculated in transgenic and non-transgenic plants (control) for the infectivity assay. The transgenic plants containing Cas12a-MV showed rare symptoms and remained healthy compared to control plants with severe symptoms. The transgenic plants containing Cas12a-MV showed a significant reduction in virus accumulation (0.05) as compared to control plants (1.0). The results demonstrated the potential use of the multiplex LbCas12a system to develop virus resistance in model and crop plants against begomoviruses.

Oleoresins are major coloring and flavoring components of herbs and spices. Among spices, garlic is a major spice used widely in cuisines and other food formulations all around the world. Garlic oleoresins have a promising future as functional ingredient in food industries. Apart from conventional extraction methods, by using modern extraction techniques, a higher yield of oleoresins can be achieved without affecting their phytochemicals, antioxidants and other characteristics. In present research, 5 different extraction methods were used for extraction and characterization comparison. After cleaning, garlic bulbs were converted to powder and moisture was optimized to 12

Methicillin-resistant Staphylococcus aureus (MRSA) has evolved resistance even against the last resort β-lactam antibiotics. This is because of the acquisition of an additional penicillin-binding protein 2a (PBP2a) which is a resistance determinant in MRSA. Currently, available PBP2a inhibitors are ineffective against life-threatening and fatal infections caused by microorganisms. Therefore, there is an urgent need to screen natural compounds that could overpass the resistance issue alone or in combination with antibacterial drugs. We studied the interactions of different phytochemicals with PBP2a so that crosslinking of peptidoglycans could be inhibited. In structure-based drug designing, in silico approach plays a key role in determining phytochemical interactions with PBP2a. In this study, a total of 284 antimicrobial phytochemicals were screened using the molecular docking approach. The binding affinity of methicillin, -11.241 kcal/mol, was used as the threshold value. The phytochemicals having binding affinities with PBP2a stronger than methicillin were identified, and the drug-likeness properties and toxicities of the screened phytochemicals were calculated. Out of the multiple phytochemicals screened, nine were found as good inhibitors to be PBP2a, among which cyanidin, tetrandrine, cyclomorusin, lipomycin, and morusin showed strong binding potential with the receptor protein. These best-selected phytochemicals were also docked to the allosteric site of PBP2a, and most of the compounds revealed strong interactions with the allosteric site. These compounds were safe to be used as drugs because they did not show any toxicity and had good bioactivity scores. Cyanidin had the highest binding affinity (S-score of -16.061 kcal/mol) with PBP2a and with high gastrointestinal (GI) absorption. Our findings suggest that cyanidin can be used as a drug against MRSA infection either in purified form or that its structure can lead to the development of more potent anti-MRSA medicines. However, experimental studies are required to evaluate the inhibitory potential of these phytochemicals against MRSA.

Medicinal plants are a rich source of bioactive compounds and have been used for the treatment of infectious diseases. Allium cepa exhibited several pharmacological activities including antimicrobial, antioxidant, hemolytic, cytotoxic, and thrombolytic effects. However, a comprehensive understanding of its therapeutic properties is still lacking. Therefore, this study was conducted to investigate the medicinal significance and biological activities of Allium cepa. The extraction was carried out through fractions of methanol, ethanol, and distilled water. The antimicrobial activity was assessed through disc diffusion method, while antioxidant activity of extract was determined through DPPH free radical scavenging assay. Hemolytic activity was evaluated to assess the toxicity of onion extract, and thrombolytic activity was performed to examine its clot lysis potential. Our results revealed that methanolic extract of A. cepa showed high zone of inhibition (23.3 mm) than n-hexane (9.2 mm) against E. coli. Methanolic extract revealed high potential of DPPH free radical scavenging (40.6%) than n-hexane (23.3%). While on the other hand, methanolic extract showed high (10.1%) thrombolytic activity than distilled water (8.3%). N-hexane fraction showed higher hemolytic activity(45.4%) than distilled water (8.2%). The results demonstrated that the extract of Allium cepa exhibited antimicrobial, antioxidant, and thrombolytic potentials due to the presence of several antioxidants and flavonoids. This study could be used for discovering the novel bioactive compounds and also helpful for exploring the therapeutic applications of medicinal plants that could be used for biological target of emerging diseases.

CRISPR-Cas9, the “genetic scissors”, is being presaged as a revolutionary technology, having tremendous potential to create designer crops by introducing precise and targeted modifications in the genome to achieve global food security in the face of climate change and increasing population. Traditional genetic engineering relies on random and unpredictable insertion of isolated genes or foreign DNA elements into the plant genome. However, CRISPR-Cas based gene editing does not necessarily involve inserting a foreign DNA element into the plant genome from different species but introducing new traits by precisely altering the existing genes. CRISPR edited crops are touching markets, however, the world community is divided over whether these crops should be considered genetically modified (GM) or non-GM. Classification of CRISPR edited crops, especially transgene free crops as traditional GM crops, will significantly affect their future and public acceptance in some regions. Therefore, the future of the CRISPR edited crops is depending upon their regulation as GM or non-GMs, and their public perception. Here we briefly discuss how CRISPR edited crops are different from traditional genetically modified crops. In addition, we discuss different CRISPR reagents and their delivery tools to produce transgene-free CRISPR edited crops. Moreover, we also summarize the regulatory classification of CRISPR modifications and how different countries are regulating CRISPR edited crops. We summarize that the controversy of CRISPR-edited plants as GM or non-GM will continue until a universal, transparent, and scalable regulatory framework for CRISPR-edited plants will be introduced worldwide, with increased public awareness by involving all stakeholders.

The most devastating pathogens of crop plants in the world are single-standard DNA viruses (Family, Geminiviridae). A vast majority of these plants are infected by the viruses belonging to the genus Begomoviruses. Chili is widely cultivated all over the world and plays an important role in the commercial sector. However, chili production in the Indian subcontinent is affected by chili leaf curl disease (ChiLCD) where it can cause a 100% loss to the crop. The current research was conducted to comprehend the begomovirus disease complex causing leaf curling disease of chili crop in Pakistan. Chili fields were surveyed for collecting infected plants with symptoms of leaf curling and stunted growth. Genomic DNA was extracted and full-length virus molecule and related satellites were cloned from infected plants using a combination of cloning techniques PCR and rolling circle amplification. A diagnostic PCR for the begomoviruses and satellites identified the presence of a well-known tomato leaf-curl New Delhi virus in the majority of the samples. However, two symptomatic plant samples showed the infection with a different begomovirus. The sequence and phylogenetic analysis of the virus molecules isolated from these two samples revealed the presence of tomato leaf curl Palampur virus (ToLCuPaV) DNA-A and an associated chili leaf curl betasatellite (ChiLCuB) in Pakistan. The nucleotide identity analysis further revealed that the Palampur isolate of DNA-A of ToLCuPaV isolated from chili in Pakistan is 97% identical to the Indian isolate (GenBank Accession-GU253914). The unique character of this isolate was the absence of DNA-B and the presence of satellite molecule. The attempts made to amplify DNA-B component using PCR or RCA were not successful. Infectious construct of ToLCuPaV was made and tested for infectivity in Nicotiana benthamiana plants using agro-inoculation with or without the ChiLCuB. The infectivity experiments suggested that DNA-A of ToLCuPaV alone was able to induce curling of leaves in N. benthamiana. Severe leaf curling symptoms were observed when N. benthamiana plants were co-inoculated with ToLCuPaV and ChiLCuB. Thus, it is concluded that betasatellite plays a critical role in the development of severe disease symptoms in tobacco plants and ToLCuPaV reported on chili plants is associated with ChiLCB instead of DNA-B component.

Oleoresins are key flavoring and coloring ingredients in spices and herbs. Among spices turmeric is most commonly used all around the world. Turmeric oleoresins have a promising future in pharmaceutical and food processing industries as functional ingredient. Apart from conventional extraction methods by using modern extraction methods a higher yield of oleoresins can be obtained without losing characteristic integrity. In the present research turmeric oleoresins were extracted by 5 different extraction methods and were compared for extraction efficiency and characterization. Turmeric samples were dried and their moisture content was further adjusted to 12% prior to extraction. Extraction was carried out by (i) conventional solvent extraction (hexane acetone petroleum ether diethyl ether and petroleum ether) (ii) steam distillation (iii) 3-phase partitioning (iv) Supercritical fluid extraction/SCFE and (v) solvent extraction followed by cryogenic grinding. The results revealed that interestingly the highest extraction efficiency was observed in solvent extraction after cryogenic grinding (4.85%). Total phenolics content (TPC) total flavonoids content (TFC) 2 2-diphenyl-1-picrylhydrazyl (DPPH) Ferric reducing antioxidant power (FRAP) and Lipoxygenase (LOX) inhibition assay were determined through UV-vis spectrophotometer functional characterization was carried out by Fourier transformed infrared (FTIR) spectroscopy while fatty acids profiling was carried out by gas chromatography attached with flame ionization detector. Total volatiles and non-volatiles were also determined by oven drying method. Characteristically oleoresins extracted through SCFE were found to be a rich source of TPC (20.57 mg GAE g-1) TFC (109.00 mg QE 100g-1) and total volatiles (2.23%) while highest non volatiles were observed in extracts obtained after cryogenic grinding. Similarly the highest antioxidant activities assessed by DPPH and FRAP were indicated by oleoresins extracted through SCFE as 59.55% and 220.60 ?M Fe2+ g-1 while highest LOX activity (58.33%) was observed in extract obtained by 3-phase partitioning. FTIR spectroscopic analysis of the selected sample indicated the presence of different functional groups with bond type at different wave numbers. GC-FID analysis of selected sample indicated the presence of 13 different fatty acids ranging from 1.9-18.6%. Pentadecane was found in lowest percentage while cis-10 pentadecanoic acid was found in highest concentration. Overall it was concluded that although extraction % of SCF extract was slight lower (4.07%) but it was found best in term of purity and characterization with a strong potential to be used in food and other related industries.

Aims: Damage of crop plants due to pathogenic attacks, postharvest crops spoilage and lethal effects of chemical pesticides has enforced scientists to find some potential natural alternative. Plants produce different types of antimicrobial peptides including defensins in response to biotic and abiotic stresses. Defensins are small cysteine rich, cationic peptide with 40-45 amino acid residues with a variety of biological activities. Hence defensins have pharmaceutical and agricultural significance especially wide range of antifungal activities. Methodology: Nine plant species viz Brassica napus, Brassica nigra, Conyza bonariensis, Alhagi marorum, Sonchus arvensis, Brassica compestris, Eruca sativa, Cirsium arvensis and Brassica juncea were selected for the study. Four set of primers were applied on extracted genomic DNAs and four amplified genes were isolated from different plants species by PCR. The amplified genes were cloned via pTZ57R/T in E.coli. The cloned fragments were sequenced and characterized by different bioinformatics tool such as coding length and peptide sequence, site of cell aggregation, peptide physico-chemical properties, antimicrobial properties and gene expression pattern. Result: The amplified products from B. napus, B. nigra, B. compestris, E. sativa and B. juncea were about 330 bp in length and showed upto 85% nucleotide homology to the reported defensins. Deduced amino acid sequence from Bn-Def, Es-Def Bj-Def and Bc- Def showed the conserved defensin domain. These sequences were also characterized for different characteristic like charged amino acids, pI value, shelflife and stability. Sonchus arvensis didn’t give any specific product by prescribed set of primers. However the products from Alhagi marorum was about 770 bp and B. napus was of 270 bp but both give homology below 30%. Conclusion: This study showed that in silico characterization of deduced antimicrobial peptides from different plant species has depicted these as an effective alternative to communal therapeutics. However, further work is needed to verify defensins specific activity at protein level.