Clustered regularly interspaced short palindromic repeats (CRISPR) associated protein (CRISPR/Cas9) is an adaptive immune system of bacteria to counter the impending viral pathogen attack. With persistent improvements, CRISPR has become a versatile tool for developing molecular immunity against viruses in plants. In the current report, we utilized the Cas9 endonuclease and dual 20 bp-gRNAs targeting two different locations in single-stranded DNA-A of AC1 (rep protein) and AV1 (coat protein) of mungbean yellow mosaic virus for achieving resistance in greengram. The cotyledonary nodal explants were infected with Agrobacterium strain EHA105 harboring pMDC100-Cas9 with AC1 and AV1 gRNA cassettes and generated transgenic plants. The integration of Cas9 and gRNA cassettes in the transformed plants of greengram were confirmed by PCR and dot blot assays. Agroinfiltrated T₂ transgenic lines exhibited minimal mosaic symptoms. A drastic reduction in the accumulation of AC1 and AV1 was observed in T₂ transformed lines. The T7EI assay indicated that AC1 fragments were edited at a frequency of 46%, 32%, 20%, and AV1 at 38.15%, 40%, and 21.36% in MYMV infected greengram lines T₂-6-2-3, T₂-6-4-4, and T₂-6-4-7, respectively. The manipulation of resistance to MYMV through the editing of the pathogen genome using the CRISPR/Cas9 tool can be a powerful approach to combat viruses and develop resistance in greengram.

Withanias omnifera also known as Indian ginseng is commonly found in India and other Southeast Asian countries. Various parts of this plant have been used as herbal medicine to treat a variety of diseases. However, there is a lacuna in the profiling of phytochemical constituents present in the different parts of the plant at reproductive stage. To identify phytochemicals present in the methanolic extracts of leaf, root, and stem parts of W. somnifera at reproductive stage using GC-MS analysis. Methods: The airdried parts of plant (leaf, stem and root) were extracted with methanol and concentrated under reduced pressure at 40°C using a rotary evaporator. The GCMSQP2010, Shimadzu, Kyoto, Japan with headspace sampler (AOC-20s) and autoinjector (AOC-20i), was used for sample analysis. The phytochemicals were identified with the database provided by National Institute Standard and Technology (NIST11LIB). The GC-MS analysis of leaf, root, and stem methanolic extracts of W. somnifera, revealed a total of eighty-two unique phytochemical peaks in the reproductive stage of the plant. Phytochemicals with antimicrobial and anticancer properties were identified in all the parts. In leaf, 2-pentanone, 5-chloro- was found to be most abundant and 2,5-dimethoxy-4-propoxy-. beta.-methyl-.beta.-nitrostyrene least abundant with antimicrobial nature, whereas, benzene, 1,1'-(1,2-ethenediyl)bis[2-methyl- was found to be most abundant and dibenzo[a,e]cyclooctene, 5,6,11,12-tetrahydro- least with anticancer property. In roots, the most abundant was benzoic acid, 3-methyl-2-trimethylsilyloxy-, trimethylsilyl ester and tris(trimethylsilyl)hydroxylamine the least abundant were identified to be antimicrobial, whereas high abundance uleine and low abundance 2-{4-[2-(4-methoxymethylphenyl)vinyl]phenyl}propan-2-olwere identified to be anticancer. In stem, acetohydroxamic acid was found to be most abundant and trans2,3,6-trimethoxy-b-methyl-b-nitrostyrene least abundant for antimicrobial nature, whereas 3-acetoxy-2,3'-bibenzo[b]thiophene was found to be anticancer phytochemical. In this study, phytochemicals with antimicrobial and anticancer properties were identified in leaf, root and stem parts of W. somnifera at reproductive stage.

Withania somnifera has been used for a long time in traditional medicine. Its crude extract, dried powder, and purified metabolites from mature plants have shown promising therapeutic potential. To further investigate its potential, the detection of phytochemicals with anticancer and antimicrobial properties in the vegetative stage is essential. Hence, this study was done to identify phytochemical constituents using GC-MS analysis for anticancer and antimicrobial activities in the vegetative stage from methanolic extracts of stem, leaf, and root in W. somnifera. The air-dried plant parts were extracted with methanol at low pressure to concentrate using a rotary evaporator at 40°C. To identify phytochemicals, Shimadzu GCMSQP2010, Japan, was used with the NIST107.LIB database. The GC-MS identified 35 unique phytochemical peaks at the vegetative stage in W. somnifera. In leaves, the antibacterial phytochemicals included cyclotrisiloxane, hexamethyl, with a high abundance, and cyclohexasiloxane, dodecamethyl, with the least abundance. In roots, the phytochemicals 2,2-dimethoxybutane, with high abundance, and cathinone, with least abundance, were found to have antibacterial properties, whereas trans-2,3-epoxyoctane, with high abundance, and 2,2-dimethoxybutane, with least abundance, were found to have anticancer properties. In stem, the antibacterial phytoconstituents octasiloxane, 1,1,3,3,5,5,7,7,9,9,11,11,13,13,15,15-hexadecamethyl, and benzenemethanol, alpha.-(1-aminoethyl), were found to be the most abundant and least abundant, respectively, while arabinitol and pentaacetate had both anticancer and antibacterial activities. At the vegetative stage, GC-MS studies of stem, leaf, and root parts revealed the occurrence of potential phytochemicals for antibacterial and anticancer activities in W. somnifera.

Accreditation is the process which ensures that certification practices are implemented in laboratories to enhance their quality and efficiency. It in turn helps laboratories to improve technical processes, achieve competitive advantage and increase market share. To achieve accreditation, successful implementation of the laboratory quality management system (LQMS) is a requisite. In this study, evaluation of quality system implementation in small, medium and large sized laboratories, covering management and technical requirements were carried out. The study analysis was carried out by scoring the implementation of quality system in various operational activities of laboratory system. Data was gathered by auditing the laboratories using check list for the purpose of International Organization for Standardization (ISO) quality management system implementation. This study, emphasize that training should be an essential element, and can play a major role in creating awareness and understanding to implement quality system in medical testing laboratory. There should be real time training on various aspects of laboratory activities. The training needs should be evidence based and assess the competency of laboratory staff, and evaluate staff performance in order to maintain world class service of the laboratory. The quality indicators can be used for benchmarking and improving services. The study conclude that LQMS in medical testing laboratories explicate the need for understanding current standard requirements of quality system implementation and maintenance to improve the quality of service of the laboratories and facilitate accreditation. A break down in implementation of quality systems can cause a decline in quality services and hence accreditation. Key words: Laboratory quality management systems, accreditation, quality indicators, implementation, training.

The prevailing abiotic stresses, especially heat stress is of great significance on the growth of plants, yield and distribution. In the conditions of heat stress, plants modulate protein processes leading to development of heat tolerance. Of such proteins, the molecular chaperone functions of HSP70/HSC70 proteins are important where their enhanced expression positively correlates with the acquisition of heat tolerance. The key players in the regulation of such tailored protein responses of plants to heat stress are the phytohormones. In the present study, phytohormone mediated interaction of Pennisetum glaucum HSC70 (PgHSC70) protein was performed through docking studies involving sequence analysis, 3D modeling and model evaluation. In silico analysis has shown better interaction and good binding energy of PgHSC70 with the phytohormone brassinolide. Furthermore, the predicted structural information can be helpful for future studies on role of interaction between HSC70 and brassinolide in heat tolerance.

Background. The ayurvedic plant Withania somnifera, a member of the Solanaceae family, has been used as a remedy for diverse health problems, including cancer. Objectives. The objective of this investigation was to conduct a comparative analysis of the in vitro cytotoxic properties of methanolic extracts derived from the leaf, stem, and root of W. somnifera on HepG2 and L929 cell lines. Methods. Methanolic extracts were obtained using the Soxhlet extraction method. To assess the in vitro anticancer action on the HepG2 and L929 cell lines, an MTT assay was performed. Changes in cell morphology were observed using an inverted microscope. Results. The MTT assay results indicated that the leaf, stem, and root methanolic extracts of W. somnifera showed significantly higher in vitro cytotoxicity in HepG2 cells, with IC₅₀ values of 43.06 ± 0.615, 45.60 ± 0.3, and 314.4 ± 0.795 μg/mL than in L929 cell lines with 78.77 ± 0.795, 90.55 ± 0.800, and 361.70 ± 0.795 μg/mL, respectively. The leaf methanolic extract was the most effective, followed by the stem methanolic extract in the HepG2 cell line. Conclusion. The results of our study have confirmed that the methanolic extracts of both the leaf and stem of W. somnifera exhibit significant in vitro cytotoxicity in HepG2 cell lines, while displaying no significant cytotoxicity in the L929 cell line. Furthermore, the data obtained from the MTT assay indicate that the leaf methanolic extract possesses a more potent cytotoxic activity than the stem methanolic extract with respect to the HepG2 cell line. Further studies on the identification and isolation of bioactive metabolites are required to explore the mechanisms underlying their in vitro cytotoxicity.