Molecular and Biological Characterization of Some Saudi Arabian Date Palm Fruits (Phoenix dactylifera L.)

تاريخ النشر (نص حر)
2015
مدى
1 item
الموضوع
نوع الرسالة الجامعية
thesis(M.A.)-King Khalid University, Faculty of Science, Department of Biology, 1437.
الملخص

Objectives: The dried fruits of date palm seven cultivars (Ajwa, Nabtat Ali, Lbanah, Anbar, Khalas, Safawi and Sukkary) were used to elucidate to the followings: 1) Physical properties 2) Possible DNA extraction form various parts 3) The DNA fingerprint patterns and genetic diversity using RAPD and ISSR primers 4) Characterizations of chemicals compounds 5) In vitro antimicrobial activities of solvent extracts of fleshy and seeds parts. Methods: 1) Colour morphology, fresh weight, dry weight, dry matter content, moisture content, perimeter, surface area and volume of dissected embryo, seed and fleshy fruit parts of seven cultivars of date palm were calculated based on weights determined before and after oven drying seed samples at 80oC for 48 hours and using mathematical formulas 2) Dried fruits were dissected into perianth, seeds without embryo, dissected embryos, endocarp, mescocarp and epicarp and the DNA was extracted from each part using commercial kit, DNeasy plant mini kit 3) Eleven RAPD primers and nine ISSR primers were used to study the variation in DNA fingerprints pattern and genetic diversity among the seven date palm cultivars 4) Antimicrobial activities of cold water, hot water, acetone, chloroform, ethanol, and petroleum ether extracts of seed and fruit of Phoenix dactylifera various cultivars were examined using 0.1 ml of standardized inoculums suspension, and using the agar diffusion method test against Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Staphylococcus aureus, Shigella sp., Klebsiella oxytoca and Candida albicans. The diameter of the zones of inhibition was measured to the nearest mm after incubation aerobically at 29°C 5) Detecting the phytochemicals and vitamins in dissected embryo, seed and fleshy fruit parts were accomplished using Gas Chromatography-Mass Spectrometry analysis and High performance liquid chromatography. Results: Obtained data showed the followings: 1) Colour morphology, fresh weight, dry weight, dry matter content, moisture content, perimeter, surface areaand volume of dissected embryo, seed and fleshy fruit significantly differed among various date palm cultivars 2) The DNA could be extracted from the dissected embryos only among other examined parts of the dried date palm fruits 3.1) One hundred eighty six bands were resulted from RAPD primers and an equal amount of polymorphic and monomorphic bands were found representing (40.47%), while numbers of unique bands (19.05%). Genetic similarity coefficients based on RAPD biomarkers showed that the highest similarity value between Lbanah cv. and Nabtat Ali cv. followed by Khalas cv. and Sukkary cv. while the least similarity value between Ajwa cv. and Safawi cultivar. Generated dendrogram separated the genotypes of seven date palm cultivars into three main clusters. The first cluster include (Ajwa cv. Anbr cv. and Khalas), the second cluster includes 4 cultivars: Lbanah cv., Nabtat Ali cv. Safawi cv. and Sukkary cultivars. 3.2) From ISSR markers, one hundred forty two of which 52 polymorphic bands, 78.0 monomorphic bands and 12.0 unique bands. Generated dendrogram separated the genotypes into three main clusters. The first cluster include (Ajwa cv. Anbr cv. and Khalas) which bifurcated into three main cluster, the first one separated Ajwa cv. from Anbr cv. and Khalas cv. The second cluster includes 3 cultivars: Lbanah cv., Nabtat Ali cv. and Sukkary cultivar. The third cluster includes Safawi cv. only. Genetic similarity showed that the highest similarity value between Anbar cv. and Khalas cv. followed by Ajwa cv. and Anbar cv. and Lbanah cv. and Nabtat Ali cv. while the least similarity value between Khalas cv. and Safawi cultivar 3.3) Based on combined analysis of RAPD and ISSR obtained data showed that the highest similarity value between Lbanah cv. and Nabtat Ali cv. followed by Anbar cv. and Khalas cv. while the least similarity value between Ajwa cv. and Safawi cultivar 4.1) Various phytochemicals and vitamins were found in dissected embryo, seed and fleshy fruit parts of each examined date palm cultivar indicating the existence of chemical compounds polymorphism 4.2) Resulted dendrogram based on chemical compounds found in embryo, seed and fleshy fruit parts could differentiae the seven cultivars of date palm in a way differed from each other 4.3) Analysis four randomised trials of vitamin in embryo, seed and fleshy fruit parts of each date palm cultivars found that each part in each examined cultivars contains a considerable amount of B1, B2, B12 and folic acid 5) Examining antimicrobial properties of solvents extracts of seed and fruit of P. dactylifera seven cultivars against Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Staphylococcus aureus, Shigella sp., Klebsiella oxytoca and Candida albicans showed that each extract from each cultivar have variable inhibiting activities on such pathogens. Conclusion: 1) The investigation the physical properties of embryo, seed and fleshy fruit parts of fruits might add precise evidence to taxonomic identity to thedate palm various cultivar 2) Simply, DNA can be extracted from the embryo of dried date palm fruits instead of difficulties to get the fresh leaves and preservation in the liquid nitrogen 3) Both ISSR and RADP markers could be effectively used in evaluation of genetic relationships among date palm various cultivars 4.1) The investigation the chemical constituents of embryo, seed and fleshy fruit parts of fruits might add precise evidence to taxonomic identity to the date palm various cultivar 4.2) Identified chemical compounds and vitamins in each part of each cultivar are biologically important that could be used in several aspect of human life especially what in the seed instead of throwing it in the junk basket 5) Solvent extracts of the fruit and seeds of date palm possess significant antimicrobial activity recommending that they could be used as a potential candidate plant for medical microbiology to prevent many human diseases