One of major limitations in the CRISPR/Cas12a-based Nucleic Acid Detection is need for T7 RNA transcription polymerase for crRNA synthesis. Here, we prepared the LbCas12a-crRNA complex, LbCo-RNP without RNA synthesis in vitro by co-expressing LbCas12a with crRNA in Eshcerichia coli. LbCo-RNP has a considerable fluorescent intensity against full matched target DNA and higher specificity against single or double mismatches in target region. We combined LbCo-RNP with PCR by consensus primer mix MY09/11 to develop the PCR-LbCo-RNP assay for human papillomavirus (HPV) genotyping. The detection limit of PCR-LbCo-RNP assay for HPV 16 or HPV 18 was 100 copies or 10 copies per reaction tube, respectively. In addition, we validated the performance of PCR-LbCo-RNP assay in 30 HPV-positive clinical samples, demonstrating that it can be widely used in clinical application.
This study investigated the development of probiotic field pea yoghurt using five lactic acid bacterial cultures—Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus acidophilus, Lactobacillus casei, and Streptococcus thermophilus, across varying inoculum concentrations (0.10–2.50
Plant endophytes confer diverse advantages to their hosts by promoting growth and development. Beneficial plant-endophyte interactions are in use for sustainable agricultural practices and for enhanced phytoremediation of environmental pollutants. Production of “high-value” secondary metabolites, with diverse therapeutic potential, by the endophytes has also gained the attention of researchers paving the way for the application of such metabolites for pharmaceutical purposes. Bioactive secondary metabolites, such as alkaloids, phenolics, steroids, quinones, tannins, saponins, terpenoids, etc., produced by the endophytes, such as Streptomyces spp., Burkholderia spp., Bacillus spp., Serratia spp., Pseudomonas spp., Paenibacillus spp., etc., have been found to possess antibacterial, antiviral, antifungal, and anticancer properties, thus, having immense potential of usage as drugs against life-threatening diseases. However, the actual potential of the endophytes in this regard is underexplored due to limited knowledge of their diversity, molecular aspects of plant-microbe interactions, and their actual functional potentials and in-planta activities. Contemporary advanced omics-based, and genetic manipulation methods have allowed researchers to better explore and utilize endophytes in drug discovery. In this review, the potential applications of bacterial endophytes as important sources of drugs against various diseases have been discussed. Recent technological advancements and prospects of research on endophytes are also reviewed.
Morphological features, cellular autofluorescence, energy and redox status of the yeast population under hyperosmolar conditions have been studied. A decrease in growth properties, the appearance of dimorphic cell forms, and changes in the surface and globularity of the cells exposed to hyperosmotic stress were shown. Assay of the cell energy status with respiration rate, cyanide resistance assessment, and staining with potentiometric probes demonstrated a decrease in the mitochondrial potential of the cells treated with NaCl. The metabolic activity of the yeast decreased by 50
Phosphate-solubilizing bacteria (PSB) are a group of microorganisms capable of mobilizing and solubilizing insoluble phosphates fixed to the soil matrix, making the nutrients readily soluble and absorbable by plants. Along with growth and yield-promoting properties, this supplementary phosphorus nutrition enhances defence systems against multiple stresses. Plant stress includes biotic and abiotic determinants. Beyond nutrient acquisition, the introduction of PSB plays a critical role in agricultural soils by effectively mitigating biotic and abiotic stresses. The strategy leads to less reliance on the use of harmful chemical fertilizers. This review provides a critical analysis of potential of PSB as a sustainable approach to reducing plant stress and enhancing agricultural resiliency. Functional diversity helps to support the efficacy of some PSB genera. The presence of PSB fits well into the organic and sustainable farming paradigm since it can be used to increase soil fertility, improve positive soil microbiomes and reduce the use of chemicals. In addition, this review covers the existing limitations to scaling up PSB as a method of stress mitigation and refocuses the opportunities of climate-resilient agriculture in future. Moreover, this review incorporates the molecular, physiological and ecological insights to relate lab-related events with field-scale applications. Although the mechanistic property of phosphate-solubilizing activity between the bacteria and the plants is not completely understood to date, certain studies on strain-specific and crop-specific use can positively impact agricultural productivity and sustainability.
