Deanship of Graduate Studies

Document Details

Document Type : Thesis 
Document Title :
Molecular and Protein Analysis for the Identification of Salt Tolerance of the In-vitro Propagated Lemon Genotypes
التحليل الجزيئي والبروتيني لتحمل الملوحة في اصناف الليمون الناتجة من الاكثار المعملي الدقيق
Subject : Faculty of Meteorology, Environment and Arid Land Agriculture 
Document Language : Arabic 
Abstract : The presented study considered the in vitro propagation using nodal segments and DNA fingerprinting of four lemon culttivars. The experiments were conducted during 2012-2014 in the Lab of Plant Tissue Culture and the Lab of Plant Biotechnology, Department of Arid Land Agriculture, Faculty of Meteorology, Environment and Arid Land Agriculture, King Abdulaziz University. The first experiment considered the effects of growth regulators on in vitro nodal segments of the lemon cultivars. The second experiment investigated the impacts of adding different growth regulators combinations at low levels to the culture medium for enhancing shoots multiplications and elongations. The third experiment investigated root induction and roots formation of the lemon formed plantlets. The fourth experiment focused on in vitro secreening of lemon cultivars to salinity tolerance. The fifth experiment focused on the DNA fingerprinting and possibly genetic diversity of the four tested lemon cultivars using RAPD-based PCR technique. The experiments of in vitro culture of Lemon were laid out in Completely Randomized Design (CRD) using 4 replicates. The results of first experiment revealed obvious significant differences due to effects of lemon cultivars, plant growth regulators combinations and their interactions on in vitro nodal segments of lemon at all measured traits. The results illustrated that the low concentrations of Kin and BAP alone or in combination with 2,4-D caused early buds sprouting of all lemon cultivars. The lemon cultivars ‘Shehri’ and ‘Aln-Eureka’ revealed highest percentages of sprouted buds on culture medium supplemented with a combinations of BAP at high concentrations and 2,4-D at low concentrations. It was observed no dead shootlets of ‘Shaary’, ‘Aln-Eurka’ and ‘Banziher’on culture medium contained low concentrations of Kin, BAP alone or in combinations with 2,4-D. The tallest primary shoots were produced by ‘Shehri’ (25.4mm) on cultutre medium contained BAP at 0.5 mgl-1+2,4-D at 1mgl-1. The lemon cultivar ‘Aln-Eureka’ produced the maxiumum number of leaves/shoot on culture medium contained BAP 0.5mgl-1 + 2,4-D at 1mgl-1. For shoot multiplication, the results showed that all explants of ‘Aln-Eureka’ developed shoots on culture medium supplemented with Kin 1mgl-1 and BAP at 0.5mgl-1 and 1mgl-1. Adding BAP at 0.5mgl-1 and 1mgl-1 to the culture medium enhanced shoots formation of the lemon cultivar ‘Aln-Eureka’ and ‘Shaary’. The greatest lengths of formed shoots were produced by ‘Shaary’ explants on culture medium supplimented with BAP at 0.5mgl-1 (39.87mm), 2mgl-1 (38.83mm). Adding BAP at 0.5mgl-1 to the culture medium increased number of formed leaves/shoot for ‘Aln-Eureka’ and ‘Shaary’, and BAP at 1mgl-1 increased number of leaves/shoots of ‘Aln-Eureka’ and ‘Banziher’. Shoots of ‘Aln-Eureka’ formed roots higher by 27.21%, 43.84% and 75.72% than ‘Shaary’, ‘Shehriu’ and ‘Banzahir’, respectively. Adding a combination of NAA at 0.5mgl-1 and IBA at 1mgl-1 estimulated higher percentages of lemon shoots to form roots (35.42%) followed by IBA at 1mgl-1 (33.33%). The cultivar ‘Aln-Eureka’ formed highest number of roots/shoot (1.76) followed by ‘Shaary’ (0.909), ‘Shehri’ (0.818) and ‘Banziher’ (0.697). The presence of NAA at 0.5mgl-1 in the culture medium significantly increased number of roots/shoot (2.55). ‘Shaary’ with 12.69 mm and ‘Aln-Eureka’ with 11.30mm produced the highest root lengths. The combination of NAA at 0.5mgl-1 and IBA at 1mgl-1 produced the highest length of roots (21.43mm). Regarding salinity tolerance the results showed that all tested cultivars revealed highest performance on medium without salt treatments (control) and lowest performance on culture medium with 0.7% and 0.9% NaCl. The cultivar ‘Banzahir’ and ‘Aln Eureka” registerd highest no. of developed shoots, no. of leaves/shoot, average shoot length (mm) and maxiumum internode length on cultutre medium with various NaCl concentrations especially 0.1% and 0.3%. The cultivar ‘Baziher’ and ‘Aln-Eureka’ revealed highest performance under various treamtemnts of NaCl and considered to be salt tolerance. ‘Banzaiher and ‘Aln Eureka’ regestered highest no. of shoots without salt stress symptoms with control treatments and 0.1% - 0.9% treatments of NaCl. The cultivars ‘Shehri’ and ‘Shaary’ recoreded high values of number of soots with mild chronics, medium chlornic and intense damaged on all culture medium with 0.1%-0.9% NaCl concentrations. These cultivars considered as salt sensitive. About the identification of lemon cultivars using RAPD markers, the results revealed that 35 (58.33%) reproducible RAPD primers out of 60 produced a total of 128 bands (monomorphic and polymorphic bands) with an average 3.65 bands/primer. Out of the 35 RAPD primers 11 were informative and produced a total of 51 bands where 29 were monomorphic (56.86%) and 22 (44.00%) polymorphic. The lemon cultivar ‘Aln-Eureka’ was identified by 14 RAPD loci, ‘Banziher’ by 16 RAPD loci, ‘Shehri’ by 10 RAPD loci and ‘Shaary’ by 16 RAPD loci. The RAPD marker OPT-03-400 was tightly linked to the lemon cultivars ‘Banziher’ and ‘Shaary’, while the marker not identified ‘Alin-Eurka’ and ‘Shehri’. OPT-06-400 was marker specific for ‘Shehri’ and ‘Shaary’, while OPT-06-450 was tightly linked to ‘Banziher’ and considered as marker specific for the lemon cultivar ‘Banziher’. Less similarity was observed between ‘Alin-Eureka’ and ‘Banziher’ (0.066), ‘Shehri’ (0.083) and ‘Shaary (0.066). High similarities were found between ‘Shehri’ and ‘Shaary’ (0.231), Banziher’ and ‘Shaary’ (0.218) and between ‘Banziher’ and ‘Shehri’ (0.153) 
Supervisor : Dr. Magdi Ali Ahmed Mousa 
Thesis Type : Doctorate Thesis 
Publishing Year : 1438 AH
2017 AD
Co-Supervisor : Dr. Ahmed Abdullah Bakhashwain 
Added Date : Thursday, January 12, 2017 


Researcher Name (Arabic)Researcher Name (English)Researcher TypeDr GradeEmail
عبدالمحسن ردة السيدAl Sayed, AbdulMohsin RadahResearcherDoctorate 


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