Biochemical Study and Molecular Approaching on Eight Wild Herbals Used in Folk Medicine in Al-Baha Region, KSA

studies   describing   the   traditional   therapeutic   uses   of   natural   medicinal   and   aromatic   plants   have been established   in   Al-Baha   region, KSA.   In   this work, the antioxidant   activity   of   eight   herbals   plant   samples   from   Al-Baha   region   was   studied   using   different   solvents (ethanol, hexane   and water) to   evaluate   their   strong   and   total   lipid-soluble   antioxidant   and   water-soluble   antioxidant   activity (TLAC37,95 and TWAC37,95). The eight herbal plants were also molecularly   characterized   using   the   PCR-based   ISSR   markers. The studied samples demonstrated   valuable   and   variable   antioxidant   properties.   The   strong   antioxidant   capacity   of   Rumex   (Rumex   nervosus Vahl.)   samples   for   ethanolic   and   aqueous   extract   samples   was   the   highest   followed by Fringed Rue (Ruta chalepensis L.) samples (47.68 and 42.72 & 81.74 and 74.18   µmoles/g   respectively),   while   hexanic   extracts   of   Euryops   (Euryops   arabicus Steud. ex Jaub. & Spach) samples   revealed   the   highest   strong   antioxidant   capacity   followed   by   Fringed   Rue   samples   (23.67   and   13.83   µmoles/g   respectively).   Besides   that, ethanolic   extracts   of   Fringed   Rue   samples   demonstrated   the   highest   total   antioxidant   capacity   followed   by   olive   leaves (Olea europaea subsp. cuspidata (Wall. & G.Don) Cif.) samples (686.95 and 462.62 µmoles/g respectively), while aqueous   extracts   of   Rumex   samples   revealed   superior   total   antioxidant   capacity   followed   by   olive   leaves   samples   and   Fringed Rue samples (869.10, 837.21   and   743.75   µmoles/g   respectively.   But   the   hexanic   extracts   of   Fringed   Rue samples   revealed   valuable   total   antioxidant   capacity   followed by Rumex samples (127.83 and 38.00 µmoles/g respectively.   The   herbal   plants   of   Rumex, Fringed   Rue   and   olive revealed   huge   both   total   and   strong   antioxidant   activity.   In   the   present   study we began   an   initial   approach   to   authenticate   and   identify   different   plant species   in   our   study   using   inter-simple   sequence   repeats (ISSRs) molecular markers

Molecular generation and characterization of an efficient recombinant vaccine for avian influenza A/H5N8 in Saudi Arabia

Purpose: To characterize a highly pathogenic avian influenza (HPAI) H5N8 for engineering recombinant 6-+ 2 vaccine strain based on reverse genetic technology. Methods: A total of 135 swab samples from various birds were collected from different parts of Saudi Arabia as part of an influenza surveillance activity. The samples were checked for influenza virus infection using reverse transcriptase-polymerase chain reaction (RT-PCR). Furthermore, Avian influenza H5N8 (A/chicken/KSA/1-NRC/2018), was used for the generation of H5N8 vaccine strain. The vaccine was tested on specific pathogen-free (SPF) chicken purchased from a local market. Results: The results indicate that the candidate vaccine (rgH5N8/KSA) induced specific neutralizing antibodies in chicken, and thereby protected the chickens from subsequent infections of H5N8. Conclusion: The study reinforces the development of a vaccine against avian influenza H5N8 virus isolated in Saudi Arabia, suggesting its possible application against the influenza virus associated with bird fl

Transcriptional analysis of Rhazya stricta in response to jasmonic acid

Background: Jasmonic acid (JA) is a signal transducer molecule that plays an important role in plant development and stress response; it can also efficiently stimulate secondary metabolism in plant cells. Results: RNA-Seq technology was applied to identify differentially expressed genes and study the time course of gene expression in Rhazya stricta in response to JA. Of more than 288 million total reads, approximately 27% were mapped to genes in the reference genome. Genes involved during the secondary metabolite pathways were up- or downregulated when treated with JA in R. stricta. Functional annotation and pathway analysis of all up- and downregulated genes identified many biological processes and molecular functions. Jasmonic acid biosynthetic, cell wall organization, and chlorophyll metabolic processes were upregulated at days 2, 6, and 12, respectively. Similarly, the molecular functions of calcium-transporting ATPase activity, ADP binding, and protein kinase activity were also upregulated at days 2, 6, and 12, respectively. Time-dependent transcriptional gene expression analysis showed that JA can induce signaling in the phenylpropanoid and aromatic acid pathways. These pathways are responsible for the production of secondary metabolites, which are essential for the development and environmental defense mechanism of R. stricta during stress conditions. Conclusions: Our results suggested that genes involved in flavonoid biosynthesis and aromatic acid synthesis pathways were upregulated during JA stress. However, monoterpenoid indole alkaloid (MIA) was unaffected by JA treatment. Hence, we can postulate that JA plays an important role in R. stricta during plant development and environmental stress conditions. How to cite: Hajrah, NH, Rabah SO, Alghamdi MK, et al. Transcriptional analysis of Rhazya stricta in response to jasmonic acid. Electron J Biotechnol 2021;50. https://doi.org/10.1016/j.ejbt.2021.01.00

Isolation, characterization, and fingerprinting of some multidrug resistance clinical isolates from patients in Al-Qaseem Hospitals, Saudi Arabia

The aim of this study was to investigate the prevalence of antibiotic resistance genes in bacteria isolated from Saudi Arabian patients in the Al-Qassim region. We also examine the application of Random Amplified Polymorphic DNA- Polymerase chain reaction (RAPD-PCR) in genetic diversity of the strains in use. In this study, we have used 29 different clinical samples from Al-Qassim hospitals which consists of wound swabs, blood, sputum and urine. Clinical samples were streaked onto sterile Petri dishes containing nutrient agar media using sterile dry swabs. Additionally antibiotic sensitivity tests, micro scan walkway susceptibility tests and extraction of genomic DNA, amplification of RAPD and Inter simple sequence repeat assay were performed in this study. This study results confirmed from the clinical samples were streaked onto sterile Petri dishes containing nutrient agar media using sterile dry swabs. The PCR analysis in ISSR was confirmed that 30–70% of E. coli in Asia and Africa carried ESBLs in the GLASS 2020 study, which is on par with the 59% in VINARES 2016–2017. Comparing VINARES 2016–2017 to other nations, the prevalence of ESBL carriage among K. pneumoniae was 35%. However, RAPD results showed that all employed strains and primers could successfully fingerprint DNA. Between isolated bacterial strains, similarity dendrograms were produced. The technique generated bands with the same intensity as the typical PCR carried out using pure DNA, and it worked for all 29 bacterial strains examined. In conclusion, the results show that all of the isolates have at least one antibiotic-resistance gene, and that the PCR approach is a quick, easy, and reliable way to identify genes

Whole mitochondrial and plastid genome SNP analysis of nine date palm cultivars reveals plastid heteroplasmy and close phylogenetic relationships among cultivars.

Date palm is a very important crop in western Asia and northern Africa, and it is the oldest domesticated fruit tree with archaeological records dating back 5000 years. The huge economic value of this crop has generated considerable interest in breeding programs to enhance production of dates. One of the major limitations of these efforts is the uncertainty regarding the number of date palm cultivars, which are currently based on fruit shape, size, color, and taste. Whole mitochondrial and plastid genome sequences were utilized to examine single nucleotide polymorphisms (SNPs) of date palms to evaluate the efficacy of this approach for molecular characterization of cultivars. Mitochondrial and plastid genomes of nine Saudi Arabian cultivars were sequenced. For each species about 60 million 100 bp paired-end reads were generated from total genomic DNA using the Illumina HiSeq 2000 platform. For each cultivar, sequences were aligned separately to the published date palm plastid and mitochondrial reference genomes, and SNPs were identified. The results identified cultivar-specific SNPs for eight of the nine cultivars. Two previous SNP analyses of mitochondrial and plastid genomes identified substantial intra-cultivar ( = intra-varietal) polymorphisms in organellar genomes but these studies did not properly take into account the fact that nearly half of the plastid genome has been integrated into the mitochondrial genome. Filtering all sequencing reads that mapped to both organellar genomes nearly eliminated mitochondrial heteroplasmy but all plastid SNPs remained heteroplasmic. This investigation provides valuable insights into how to deal with interorganellar DNA transfer in performing SNP analyses from total genomic DNA. The results confirm recent suggestions that plastid heteroplasmy is much more common than previously thought. Finally, low levels of sequence variation in plastid and mitochondrial genomes argue for using nuclear SNPs for molecular characterization of date palm cultivars

Plastid SNPs sorted by position in the genome.

<p>SNPs that are unique to an individual cultivar are in bold. IGS  =  intergenic spacer; I  =  intron; NS  =  nonsynonymous; S  =  synonymous.</p

Maximum likelihood trees of mitochondrial SNPs for 10 date palm cultivars.

<p>(A), plastid (B), and combined (C). Numbers below each node represent bootstrap values for 1000 replicates. Cultivar abbreviations are provided in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0094158#pone-0094158-t001" target="_blank">Table 1</a>. Cultivar acronyms in red and black are female and male plants, respectively. Fruit shape is indicated and acronym names are color coded by fruit color (yellow, red, and brown).</p