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المستخلص This experiment was carried out during two successive seasons 2020-2021 to evaluate four avocado cultivars namely (Hass, Bacon, Fuerte, Red), were selected for physical and chemical characterizing. The experimental trees with more than twelve years old, budded on balady rootstock, grown in a sandy soil under drip irrigation system and spaced at 6x7 m apart in Nobbaria region in EL-Salmayai farmhouse, Elbehera governorate, Egypt. The present investigation aimed to: 1. Evaluating leaf and fruit physical characteristics and the fruit chemical constituents of the studied cultivars in order to identify the superior ones.2. 2. Studying the relationships among these avocado cultivars (Hass, Bacon, Fuerte, Red) using the molecular markers. 3. Fingerprinting for the extracted DNA from the leaves of the examined cultivars and Construction the phylogentic tree between the examined cultivars based on the difference obtained by the previous molecular techniques. The results of the present study could be summarized in both season as follows: - The obtained data present the highest value of the petiole length was recorded by the Hass cultivar, while the lowest was found in the Bacon cultivar in 2020 season. In 2021 season the highest value of the petiole length was recorded by the Hass cultivar, while the lowest was found in the Bacon cultivar. - The obtained data present the highest value of the blade length was recorded by the Hass cultivar, while the lowest was found in the Fuerte cultivar in 2020 season. In 2021 season the highest value of the blade length was recorded by the Bacon cultivar, while the lowest was found in the Fuerte cultivar. 116 - The obtained data present the highest value of the blade width was recorded by the Fuerte cultivar, while the lowest was found in the Bacon cultivar in 2020 season. In 2021 season the highest value of the blade width was recorded by the Bacon cultivar, while the lowest was found in the Fuerte cultivar. - The obtained data present the highest value of the Blade length\blade width was recorded by the Red cultivar, while the lowest was found in the Fuerte cultivar in 2020 season. . In 2021 season the highest value of the Blade length\blade width was recorded by the Red cultivar, while the lowest was found in the Fuerte cultivar. - The obtained data present the highest value of the Fruit length was recorded by the Fuerte cultivar, while the lowest was found in the Hass cultivar in 2020 season. In 2021 season the highest value of the Fruit length was recorded by the Fuerte cultivar, while the lowest was found in the Hass cultivar. - The obtained data present the highest value of the fruit diameter was recorded by the Red cultivar, while the lowest was found in the Hass cultivar in 2020 season. In 2021 season the highest value of the fruit diameter was recorded by the Red cultivar, while the lowest was found in the Hass cultivar. - The obtained data present the highest value of the Fruit length/ diameter ratio was recorded by the Bacon cultivar, while the lowest was found in the Red cultivar in 2020 season. In 2021 season the highest value of the Fruit length/ diameter ratio was recorded by the Fuerte cultivar, while the lowest was found in the Red cultivar. - The obtained data present the highest value of the Fruit weight was recorded by the Red cultivar, while the lowest was found in the Hass cultivar in 2020 season. In 2021 season the highest value of the Fruit weight was recorded by the Red cultivar, while the lowest was found in the Hass cultivar. 117 - The obtained data present the highest value of the flesh weight was recorded by the Red cultivar, while the lowest was found in the Hass cultivar in 2020 season. In 2021 season the highest value of the flesh weight was recorded by the Red cultivar, while the lowest was found in the Hass cultivar. - The obtained data present the highest value of the seed weight was recorded by the Red cultivar, while the lowest was found in the Hass cultivar in 2020 season. In 2021 season the highest value of the seed weight was recorded by the Bacon cultivar, while the lowest was found in the Hass cultivar. - The obtained data present the highest value of the Seed weight\ flesh weight ratio was recorded by the Bacon cultivar, while the lowest was found in the Red cultivar in 2020 season. In 2021 season the highest value of the Seed weight\ flesh weight ratio was recorded by the Bacon cultivar, while the lowest was found in the Red cultivar. - The obtained data present the highest value of the Peel weight was recorded by the Red cultivar, while the lowest was found in the Hass cultivar in 2020 season. - The obtained data present the highest value of the Peel weight was recorded by the Red cultivar, while the lowest was found in the Hass cultivar in 2020 season. In 2021 season the highest value of the Peel weight was recorded by the Red cultivar, while the lowest was found in the Hass cultivar. - The obtained data present the highest value of the moisture was recorded by the Fuerte cultivar, while the lowest was found in the Red cultivar in 2020 season. In 2021 season the highest value of the moisture was recorded by the Red cultivar, while the lowest was found in the Hass cultivar. - The obtained data present the highest value of the lipids was recorded by the Hass cultivar, while the lowest was found in the Red cultivar in 2020 season. . In 2021 118 season the highest value of the lipids was recorded by the Hass cultivar, while the lowest was found in the Fuerte cultivar. - The obtained data showed the highest value of the protein was recorded by the Bacon cultivar, while the lowest was found in the Red cultivar in 2020. In 2021 the obtained data present the highest value of the protein was recorded by the Hass cultivar, while the lowest was found in the Red cultivar. - The obtained data showed the highest value of the ash was recorded by the Fuerte cultivar, while the lowest was found in the Red cultivar in 2020. In 2021 the obtained data present the highest value of the ash was recorded by the Fuerte cultivar, while the lowest was found in the Red cultivar. - The results showed that twenty-two of the primers assisted in identifying the samples, whereas the remaining eight primers produced limited results. The 22 effective primers chosen (3, 6, 9, 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 24, 25, 26, 27, 28, 29 and 30) gave 334 amplified detectable bands in total with an average of about 23.35 bands per primer, ranging from 1500-100 bp. Primer No. 20 had the highest total number of different PCR bands (27 bands), whereas primers No. three and six showed the lowest total number of bands (four bands). However, the results provide that primer No. 12 had the highest polymorphic number of different PCR bands (six bands), whereas primers No. 9, 18, 20, and 26 showed lowest polymorphic number of different PCR bands (one band), in all results . - The relationships within and between groups were estimated by a UPGMA cluster analysis of GS matrices. Samples Red and Hass were more closely related with each other more than the Fleurte and Bacon. - Results showed the success of PCR amplification for rbcL. The rates of DNA sequencing indicated that rbcL was universal for avocado species, and that 119 consolidating this marker would improve the precision of species differentiation. The purified DNA was sequenced using the automated sequencer of the Sanger method by Macrogene Company (Korea). DNA homology searches were carried out with the National Center for Biotechnology Information (NCBI) databases, using the basic local alignment search tool (BLAST) network service. (www.ncbi.nlm.nih.gov/BLAST).However, the nucleotide sequences were illustrated |