الفهرس 
Phylogenetic relationship among different cultivars of vinca (Catharanthus roseus) plant using molecular markersOnly 14 pages are availabe for public view
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Abstract
The present investigation was carried out to study genetic diversity and phylogenetic relationships between unnamed five different Catharanthus roseus cultivars (C1, C2, C3, C4 and C5) are different from each other in colour of the flowers and grown in some Egyptian gardens, which C1 and C2 are local cultivars while C3, C4 and C5 are imported cultivars from Holland by using Randoμly Amplified Polymorphic DNA (RAPD) and Inter - Simple Sequence Repeats (ISSR) techniques. For this purpose 11 random primers and five ISSR primers were used to amplify random and certain DNA sequence respectively throughout the genome of the five different Catharanthus roseus cultivars under this study . DNA was extracted from leaves of each one of the previous cultivars by using Doyle and Doyle (1987) method then stored at -20 °C until subsequent analysis. The obtained results can be summarized in the following points:
1. The all 11 random primers produced a total of 66 amplified bands which 53 of them were polymorphic giving 8.83 % polymorphism between the five different Catharanthus roseus cultivars. The average of amplified bands per primer was 6 bands which had different base pair in their length.
2. The random primers P6A-03, OPS-12 and OPS-13 produced the highest number (8 bands) of amplified bands while the random primers OPD-01, OPB-11 and OPB-07 produced the lowest number of polymorphic bands (4,4 and 3 bands respectively).
3. The highest percentage of polymorphism (100 %) was produced by each one of the random primers OPD-11 and OPB- 05, while the lowest percentage of polymorphism (50 % and 66.66 %) were generated by the random primers OPS-11 and OPB-07 respectively.
4. Each one of the 11 random primers produced either one unique amplified band or two unique amplified bands which had different base pair in length present in the genomic of one cultivar only and absent in other cultivars and can be used as molecular markers to identify certain cultivar from others.
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5. Genetic similarity (GS) between the five different C. roseus generated from RAPD markers varied between 34 % and 57.96 % and the minimum GS value was between the local C1 and imported C3 cultivars, while the maximum GS value was between imported C4 and imported C5 cultivars.
6. The phylogenic tree constructed using the all 11 random primers showed that all the five different C. roseus cultivars were classified into to two clear clusters. The first cluster was represented by local C1 and local C2 cultivars, while the second cluster was divided into two nodes; one of them comprised imported C3 cultivar and the other node represented by imported C4 and imported C5 cultivars.
7. The highest genetic similarity revealed by the RAPD’s analysis was 57.69 % between imported C4 and C5 cultivars flowed by 50 % between imported C3 and C5). on the other hand, the percentage of GS between local C1 and C2 cultivars was 53.65%, while the lowest GS between the local C1 and imported C3, C5 and C4 cultivars were 34 04%, 38.00% and 43.13% respectively.
8. All five different ISSR primers produced a total of 21 amplified bands which 19 of them were polymorphic bands present in some cultivars and absent in others giving 90.74% polymorphism. The average of amplified bands per primer was 4.2 bands which had different base pair in their length.
9. The highest number of amplified bands (6 bands) were produced by ISSR primers HB-8 and HB-010, while the lowest number of amplified bands (2 bands) were produced by HB-09 and HB-13.
10. The highest percentage of polymorphism (100 %) was produced by each one of the ISSR primers HB-09, HB-010 and HB-012, while the lowest percentage of polymorphism (66.66 %) was generated by the ISSR primer HB-08.
11. The ISSR primers HB- 010, HB-08 and HB-012 produced different unique amplified bands which had different base pair in their length ( 1, 2, and 4 bands respectively ) present in one cultivar and absent from others. These unique amplified bands were considered as molecular markers to identify certain cultivars from others Catharanthus roseus cultivars under this study.
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12. Genetic similarity (GS) values generated from ISSR markers varied between 33 % and 63.63 % and the minimum GS value was between the local C1 and imported imported C3 cultivars, while the maximum GS value was between imported C4 and C5 cultivars.
13. The phylogenetic tree constructed from ISSR data revealed that there are two clear clusters. The first cluster was represented by imported C5 cultivar only and the second cluster was divided into two sub cluster, one of them comprised imported C4 cultivar and the other sub cluster was divided into two nodes, one of them contains local C2 cultivar and the other node was represented by local C1 and imported C3 cultivars
14. Genetic similarity resulting from the combined DNA markers data (RAPD and ISSR data ) revealed that the highest percentage of similarity (50.68 %) was detected between imported C4 and C5 cultivars followed by (44.28 %) between local C1 and imported C4 and (41.28 %) between local C1 and imported C5 cultivars.
15. It may be concluded that the obtained results from RAPD and ISSR analysis as well as combined RAPD and ISSR analysis were relatively similar considering genetic similarity between the five C. roseus cultivars.
16. The phylogenetic tree built on the basis of combined data from RAPD and ISSR analysis demonstrated that the five Catharanthus roseus cultivars were divided into two clusters, the first cluster containing local C1 and C2 cultivars, while the second cluster contains two nodes. The first node comprised imported C3 cultivar and the second node contains imported C4 and C5 cultivars.
17. It may be concluded that the all the cultivars (Different genotypes) belonging to a particular geographic region were grouped together in the phylogenic tree. It can be explained from the observation that geographically isolated individuals tend to accumulate genetic variations during environmental adaptations.
18. In conclusion, the use of RAPD and ISSR could be a powerful tool to assess the genetic diversity in Catharanthus roseus.
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19. This study provided new information at the molecular level for the genetic diversity and phylogenetic relationship among the five Catharanthus roseus cultivars. The polymorphism data generated can be used for plant breeding, varietal improvement programs and also might be helpful in future strategies for evolution of desired genotypes and further development of new C. roseus cultivars.