التوصيف الجزيئي لنبات إكليل الجبل Rosmarinus spp. واختبار الفعالية الحيوية لبعض مستخلصاته
Molecular Characterization of the Rosemary Plant Rosmarinus spp. and Testing the Biological Efficacy of some of its Extracts
- الباحث
- سكينة مدحت علي الجنابي
- المشرف
- أ.د. نضال عبد الحسين البديري
- الجامعة
- جامعة الكوفة
- الكلية / القسم
- التربية للبنات / علوم الحياة
- الشهادة
- ماجستير
- السنة
- 2022
- لغة الرسالة
- عربي
- الكلمات المفتاحية
- إكليل الجبل;التوصيف الجزيئي;المستخلصات النباتية;الفعالية الحيوية
الملخص
أجري البحث في كلية التربية للبنات / جامعة الكوفة للفترة من 2021/1/10 لغاية 2022/1/10 إذ هدفت الدراسة إلى التشخيص الجزيئي لنبات إكليل الجبل 7 والكشف عن بعض الجينات المهمة فيه بالإضافة إلى تقييم فعاليته الحيوية في تصنيع جسيمات الفضة . النانوية وتقييم فعاليتهما ضد بعض الفطريات الممرضة أوضحت الدراسة الجزيئية و من خلال تضاعف منطقة Internal transcribed spacer (ITS) وبإستعمال البوادئن 1151 و 1154 إذ Gin نتائج التضاعف بوساطة تقنية تفاعل البلمرة المتسلسل Polymerase chain reaction (PCR) وبإستعمال البوادئ المحددة والترحيل الكهربائي لنواتج التضاعف ظهور حزم عند bp 700 تقريباً , كما كشفت نتائج مقارنة تسلسلات القواعد النتروجينية للنبات قيد الدراسة مع تسلسلات القواعد النتروجينية المسجلة عالميا في المركز الوطني لمعلومات التقانات الحيوية National Center for Biotechnology )NCBI Information ) إذ أثبتت نتائج التحليل في NCBI انها تتطابق مع التسلسلات المسجلة لنبات 165 :171711 المسجل في المانيا calls يحمل الرقم التعريفي ID:_KJ584197.1 وبنسبة تطابق 799 كما بينت نتائج المحاذاة وجود ثلاث تغايرات (طفرات ) في التسلسلات قيد الدراسة وسّجلت هذه التسلسلات في Gs5 NCBI رقم إنضمام تعريفي 01/920532.1 :110 كما Gin الدراسة ومن خلال الكشف الجزيئي بتقنية PCR وجود الجينات ،2 cineole pinene synthase s synthase في أوراق نبات إكليل الجبل .
Abstract
The research was carried out at the College of Education for Girls/University of Kufa for the period from 10/1/2021 to 10/1/2022, as the study aimed at the molecular diagnosis of the rosemary plant Rosmarinus officinalis and the detection of some important genes in it, in addition to evaluating its biological effectiveness in the manufacture of silver nanoparticles and the evaluation of its efficacy against some pathogenic fungi. The molecular study showed, through the doubling of the internal transcribed spacer (ITS) region and using the primers ITS1 and ITS4, that the results of the doubling mediated by the polymerase chain reaction (PCR) technique and using the specific primers and the electrophoresis of the replication products showed the appearance of bundles at approximately 700 bp, and also revealed that The results of comparing the sequences of nitrogenous bases of the plant under study with the sequences of nitrogenous bases registered globally in the National Center for Biotechnology Information (NCBI) , The results of the NCBI analysis demonstrated that they matched the recorded sequences of the plant Rosmarinus officinalis registered in Germany, which has the ID number KJ584197.1, with a 99% certainty.The alignment results also showed the presence of three variations (mutations) in the sequences under study. OM920532.1 is the membership ID. The study also showed, through molecular detection by PCR technique, the presence of the genes cineole synthase and pinene synthase in the rosemary plant. The results of the examination of the cold ethanolic alcohol extract (99.99%) of rosemary leaves by the GC-Mass (Gas Chromatography) technique revealed the presence of 15 compounds, and the main compounds with the highest percentages were: 4,6-Bis (1,1'-dimethylethyl)-2',5'-dimethoxy-1,1'-biphenyl-2-ol by 28.38%, Caryophyllene by 13.82% and Eucalyptol by 12.46% . The study also demonstrated the possibility of manufacturing silver nanoparticles from the extract of rosemary leaves, as they appeared to have a semi-pyramidal shape with sizes ranging between 30 and 94 nm when examined by a Scanning Electron Microscope and at an average size of 63.65 nm. The appearance of an absorption peak at a wavelength of 345 nm The results of the X-ray diffraction spectrum examination also showed the appearance of four clear diffraction peaks at 2 values: 38.12, 44.32, 64.44, and 77.44, which corresponded to 111, 200, 220, and 311, respectively, from the standard scheme. (Standard) for XRD spectroscopy of silver nanoparticles (JCPDS silver: 04-0783). As for the examination by the Atomic Force Microscope, the results showed that the average size of the surface topography was 29.65 nanometers. The results of evaluating the cold alcoholic extract of rosemary leaves in the inhibition of some pathogenic fungi showed that there were significant differences between the percentages of inhibition, as the percentages of inhibition increased clearly with the increase in concentrations. The study proved when using concentrations (500 mg, 1 g, and 2 g)/L that the concentration 2 g/L showed good inhibition rates that reached 81.1% with Aspergillus niger and 86.6% with Fusarium verticillioides, while the highest inhibition was with Fusarium solani, with an inhibition rate of 87.7%. The results of inhibiting the types of pathogenic fungi studied by silver nanoparticles made from the hot aqueous extract of rosemary leaves showed significant differences and high inhibition efficiency with increasing concentrations. The concentration of ppm/liter showed high inhibition efficiency with all studied fungal species. The percentages of inhibition reached 100% with A. niger and F. solani, while the inhibition percentage for the same concentration with F. verticillioides was 73%.
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