The use of random amplified polymorphic DNA (RAPD) markers to identify a number of chickea (cicer arietinum L.)

number: 
390
English
Degree: 
Imprint: 
Biotechnology
Author: 
Yaseen Ismaeel Aumran
Supervisor: 
Dr. Jaladet M. S. Jubrael
Dr. Galib H. Al-Bakri
year: 
2002
Abstract:

The objective of this study was to detect DNA polymorphism among eleven chickpea (Ciccr airietinum L.) varieties/lines cultivated in Iraq using Random Amplified Polymorphic DNA (RAPD) Markers in order to identify different varieties / lines. To achieve this the following steps were carried out: (Genomic DNA of chickpea plants was extracted using CTAB method. The concentration of DNA recovered was ranging between (200-250) ug / 3 g of fresh plant tissues with a molecular weight of approximately 50 kbp and a purity ranging between (1.7-1.8). In this study, Twenty different random primers were evaluated for their usefulness in detecting DNA polymorphisms among chickpea genomes tested. This involved first optimization of RAPD reaction conditions including, DNA template, Taq DNA polymerase, and Primer concentrations The results obtained were as follows: Eight primers showed no amplified product across all chickpea genotypes. Twelve primers amplified genomic DNA across the chickpea varieties/lines with different efficiency. The generated DNA fragments using these primers were scored by their presence or absence and the differences in their molecular weight across the chickpea varieties/lines. The total number of amplified bands generally ranged between (21-154) and the molecular weight varied between (0.12-3.89) kbp according to the primer used. Another important result obtained from this study was the possible discrimination and / or identification of nine varieties/lines namely ( Shoki, Raffidien, Marakishi, Dijla, IPA-46,IPA-69, IPA-510, F91-52C,and F92-I20C) with seven primers that were ( OPA-07, OPA-20, OPB-09, OPE-02, OPF-02, OPG-08, OPP-12 ). In order to enhance the DNA polymorphisms among RAPD patterns produced by primer OPA-02, the amplified product was digested with the tetra nucleotide restriction enzyme Tru 91. Successful results were also obtained by detection of DNA polymorphism among patterns compared to the monomorphic RAPD patterns produced originally by this primer. The results of this experiment made also the possible identification of three different varieties/lines namely Shoki variety, I PA-510 line, and F91-52C line. These results clearly demonstrated the feasibility of using RAPD markers in breeding programs of chickpea in Iraq.