التشخيص الجزيئي لنبات Physalis angulata والتركيب الكيميائي وتقييم الفعالية التثبيطية لمستخلص أوراقه ضد بكتريا Staphylococcus aureus المعزولة من مصادر سريرية

Molecular Identification of Physalis angulata, Chemical composition and evaluation the inhibitory activity of its leaves extract against Staphylococcus aureus isolated from clinical sources

ABSTRACT

It included the molecular diagnosis of two species of Physalis spp collected in

Iraq on conservative regions, such as the sequence of chloroplast DNA (cpDNA) and the sequence of ribosomal nuclear (nrDNA) region of nuclear ribosomal DNA, obtained from one of the regions belonging to the private sector / Holy Karbala Governorate And the second residence spread naturally in some orchards, they were collected and classified according to the phenotypic form, and they belonged to the Solanaceae family, and vice versa, and from the genus Physalisen, by the operation of their self-propagation.
The results showed the isolation of the two species from each other, if the nucleotide sequences indicated the confirming identity of the amplified aggregates via the NCBI blast gene bank site. 1 with a percentage of 98.59%, which belongs to a Chinese strain of the same genus. The results also showed the presence of three mutations when comparing the sequence of nitrogenous bases. The species was registered by the researcher as Physalis peruviana isolate AH-ZE1, with serial number OQ625502.1g.
But when using nrDNA sequences, it was noted that there were high similarities of the first sample with the type of Physalis peruviana 217 deposited in the Genome Bank with the serial number DQ314161.1, with a rate of 96.91%, which belongs to a strain located in the United States of America. The results also showed that there were 8 mutations when comparing the sequence of bases. nitrogenous, and the species was recorded in the Genie Bank as Physalis peruviana isolate AH-ZE1, with serial number OQ616506.1. As for the second sample, it was noted that there were high similarities with the type Physalis angulata isolate LHR28I deposited in the Genome Bank with serial number MK412130.1, with a rate of 98.02%, which belongs to a strain found in Spain. The results also showed the presence of 10 mutations when comparing the sequence of nitrogenous bases. The type of The researcher accepted it as Physalis angulata isolate AH-ZE1 inducer, sequence number OQ616509.1.
The results of the GC-mass analysis showed that the sample Physalis angulata isolate AH-ZE1 contained twenty-four effective compounds that have a lot of medicinal efficacy. Oxidation 94% at a concentration of 140 of the extract and the lowest percentage of antioxidants 42% at a concentration of 20 is due to the presence of a high percentage of phenolic compounds in the plant extract, which are directly proportional to the percentage of antioxidants.
The results showed that the alcoholic extract of Physalis angulata AH-ZE1 was effective against bacteria the areola diameter for inhibiting Staphylococcus colonies showed that a concentration of 25% of the extract was more efficient in inhibiting Staphylococcus non-producing clotting enzyme. Inhibition of enzyme-positive Staphylococcus colonies (18.34 ± 1.24) at a significant level (P < 0.05), on the other hand, the best lowest inhibitory concentration when using the antibiotic doxycycline was 1.88 ± 42.47 mm and 1.76 ± 42.6 for both enzyme-positive and negative staphylococci colonies. Coagulation with no significant differences between antibiotics.
The results of the coa gene expression study showed that there were clear significant differences in the gene expression levels of the isolate between the treatments of the alcoholic extract of Physalis angulata and the control group, as the level of gene expression decreased in the treatment T2 with a concentration of 50%, the third T3 with a concentration of 75% and the fourth T4 with a concentration of 100%. % when compared with the first treatment with a concentration of 25%. And their comparison to the control group, that is, the concentration of 50%, 75%, and 100% negatively affects the level of gene expression, and thus affects the formation of the biofilm of Staphylococcus aureus, which leads to a reduction in bacterial growth, While the concentration of 25% of the extract showed a clear effect on the high level of gene expression of the Coa gene (an increase in the concentration of RNA), it indicates the activity and effectiveness of this gene in activating and maintaining the biofilm, which leads to an increase in the growth of pathogenic bacteria, that is, the higher the concentration of the extract, the lower it is. Gene expression of the Coa gene of Staphylococcus aureus, which indicates the efficiency of the alcoholic extract of Physalis angulata with a concentration of (50, 75 and 100)% as anti-biofilm formation.