Ehab H. El-Harty

Molecular Diversity Assessment Using Sequence Related Amplified Polymorphism (SRAP) Markers in Vicia faba L.

Sequence-related amplified polymorphism (SRAP) markers were used to assess the genetic diversity and relationship among 58 faba bean (Vicia faba L.) genotypes. Fourteen SRAP primer combinations amplified a total of 1036 differently sized well-resolved peaks (fragments), of which all were polymorphic with a 0.96 PIC value and discriminated all of the 58 faba bean genotypes. An average pairwise similarity of 21% was revealed among the genotypes ranging from 2% to 65%. At a similarity of 28%, UPGMA clustered the genotypes into three main groups comprising 78% of the genotypes. The local landraces and most of the Egyptian genotypes in addition to the Sudan genotypes were grouped in the first main cluster. The advanced breeding lines were scattered in the second and third main clusters with breeding lines from the ICARDA and genotypes introduced from Egypt. At a similarity of 47%, all the genotypes formed separated clusters with the exceptions of Hassawi 1 and Hassawi 2. Group analysis of the genotypes according to their geographic origin and type showed that the landraces were grouped according to their origin, while others were grouped according to their seed type. To our knowledge, this is the first application of SRAP markers for the assessment of genetic diversity in faba bean. Such information will be useful to determine optimal breeding strategies to allow continued progress in faba bean breeding.

Morphological and molecular genetic variability analyses of Saudi lucerne (Medicago sativa L.) landraces

Abstract. Collection and characterisation of genetic resources are required for the development of new cultivars. We analysed genetic diversity among 18 non-dormant lucerne (alfalfa, Medicago sativa L.) accessions including ten local ecotypes and eight introduced accessions at morphological and molecular levels using sequence-related amplified polymorphism (SRAP) primers. Standardised canonical discrimination functions for the investigated morphological traits showed that the first function (explaining 75% of total variability among accessions) was strongly influenced by leaflet shape, stipule shape, and the peduncle : petiole length ratio. The 16 SRAP primer pair combinations generated 677 differently sized SRAP fragments (peaks), of which 665 (98.3%) were polymorphic across all 18 accessions. We detected high levels of polymorphism (average polymorphic information content value = 0.96, average of 42.3 polymorphic fragments per primer pair). Based on morphological and SRAP data, local accessions tended to group together in the same cluster or formed individual clusters. Clusters of local accessions at high similarity sometimes correlated with their collection site (Qasemi-2 and Qasemi-3) on molecular analysis. Results of cluster analysis based on SRAP showed no significant correlation with morphological characters based on the Mantel test (r = 0.04).This inconsistent clustering of accessions could be due to the allelic variation (presumably) in a small number of genes (24 traits) contributing to morphological characterisation, while the 677 SRAP fragments (loci) are assumed to be relatively widely distributed across the genome. The wide geographical distribution of lucerne populations across different environments may provide good genetic resources for breeding purposes. SRAP analysis was effective to study genetic variability of non-dormant lucerne. This information will be helpful in assessing selections for lucerne breeding programs to develop new cultivars adapted to harsh environmental conditions.

Phenological, Nutritional and Molecular Diversity Assessment among 35 Introduced Lentil (Lens culinaris Medik.) Genotypes Grown in Saudi Arabia

Morphological, nutritional and molecular analyses were carried out to assess genetic diversity among 35 introduced lentil genotypes (Lens culinaris Medik.). The genotypes exhibited significant differences for their field parameters and some of them showed noticeable superiority. The nutritional and proximate analysis showed that some genotypes were excellent sources of proteins, essential amino acids, minerals, anti-oxidants, total phenolic contents (TPC) and total flavonoid contents (TFC) and hence, highlights lentil nutritional and medicinal potential. Sequence-related amplified polymorphism (SRAP) and amplified fragments length polymorphism (AFLP) markers were used to estimate the genetic variability at the molecular level. The existence of a considerable amount of genetic diversity among the tested lentil genotypes was also proven at the molecular level. A total of 2894 polymorphic SRAP and 1625 AFLP loci were successfully amplified using six SRAP and four AFLP primer pair combinations. Polymorphism information content (PIC) values for SRAP and AFLP markers were higher than 0.8, indicating the power and higher resolution of those marker systems in detecting molecular diversity. UPGMA (unweighted pair group method with arithmetic average) cluster analysis based on molecular data revealed large number of sub clusters among genotypes, indicating high diversity levels. The data presented here showed that FLIP2009-64L and FLIP2009-69L could be used as a significant source of yield, total protein, essential amino acids, and antioxidant properties. The results suggest potential lentil cultivation in the central region of Saudi Arabia for its nutritional and medicinal properties, as well as sustainable soil fertility crop.

Assessment of genetic diversity among faba bean genotypes using agro-morphological and molecular markers

Forty faba bean (Vicia faba L.) genotypes were evaluated for their agro-morphological performance and molecular diversity under Central Region of Saudi Arabia conditions during 2010–11 and 2011–12 seasons. Field performance results showed that faba genotypes exhibited a significant amount of variation for their agro-morphological studied parameters. Giza40 recorded the tallest genotype (139.5 cm), highest number of seeds per plants (100.8), and the highest seed yield per plant (70.8 g). The best performing genotypes were Giza40, FLIP03-014FB, Gazira1 and Goff1. Genetic variability among genotypes was determined using Sequence Related Amplified Polymorphism (SRAP) and Amplified Fragment Length Polymorphism (AFLP) markers. A total of 183 amplified fragments (alleles) and 1758 polymorphic fragments (bands) in SRAP and 202 alleles and 716 bands in AFLP were obtained using six SRAP and four AFLP primer combinations respectively. Polymorphism information content (PIC) values for AFLP and SRAP markers were higher than 0.8, indicating the existence of a considerable amount of genetic diversity among faba tested genotypes. The UPGMA based clustering of faba genotypes was largely based on origin and/or genetic background. Result of cluster analysis based on SRAP showed weak and not significant correlation while, it was highly significant based on AFLP analysis with agro-morphological characters (r = 0.01, p > 0.54 and r = 0.26, p < 0.004 respectively). Combined SRAP and AFLP markers proved to be significantly useful for genetic diversity assessment at molecular level. They exhibited high discrimination power, and were able to distinguish the faba bean genotypes with high efficiency and accuracy levels.

Comparative nutritional profiles of various faba bean and chickpea genotypes.

Grain legumes are important source of proteins and dietary minerals for humans. In this study, 40 faba bean and 28 chickpea genotypes were evaluated for their nutritional profile. We found that crude proteins ranged from 31.5-37.7% and 19.8-24.9% in faba bean and chickpea genotypes, respectively. Trypsin and chymotrypsin inhibitors varied from 2.24-2.77 and 0.35-0.70 trypsin inhibitory unit (TIU) mg -1 , respectively in faba bean genotypes; whereas, in chickpea genotypes, they ranged from 7.65-8.98 and 9.0-11.9 TIU mg -1 , respectively. Tannins in faba bean ranged from 12.2-16.2 mg 100 g -1 , while in chickpea they ranged from 4.11-4.94 mg g -1 . However, phytic acid ranged from 1.2-1.5 mg 100 g -1 in faba bean, while in chickpea it ranged from 6.10-6.98 mg g -1 . Total phenolic and total flavonoid contents ranged from 5.8-11.3 mg galic acid equivalents (GAE) g -1 and 0.08-0.16 mg quercetin equivalent (QE) g -1 , respectively in faba bean genotypes; whereas, in chickpea genotypes, these ranged from 1.5-2.5 mg GAE g -1 and 0.05-0.18 mg QE g -1 , respectively. In crux, some faba bean and chickpea genotypes could be potential donors for legume genetic improvement and incorporation of such legumes in human diets may improve the nutritional value of diet and can reduce malnutrition.

Comparative phytochemical profiling of different soybean ( Glycine max (L.) Merr) genotypes using GC–MS

This study aimed to estimate the proximate, phenolic and flavonoids contents and phytochemicals present in seeds of twenty four soybeans (Glycine max (L.) Merr) genotypes to explore their nutritional and medicinal values. Crude protein composition ranged between 35.63 and 43.13% in Argentinian and USA (Clark) genotypes, respectively. Total phenolic content varied from 1.15 to 1.77 mg GAE/g, whereas flavonoids varied from 0.68 to 2.13 mg QE/g. The GC–MS analysis resulted identification of 88 compounds categorized into aldehydes (5), ketones (13), alcohols (5), carboxylic acids (7), esters (13), alkanes (2), heterocyclic compounds (19), phenolic compound (9), sugar moiety (7) ether (4) and amide (3), one Alkene and one fatty acid ester. Indonesian genotypes (Ijen and Indo-1) had the highest phenolic compounds than others genotype having antioxidant activities, while the Australian genotype contains the maximum in esters compounds. The major phytocompounds identified in majority of genotypes were Phenol, 2,6-dimethoxy-, 2-Methoxy-4-vinylphenol, 3,5-Dimethoxyacetophenone, 1,2-cyclopentanedione and Hexadecanoic acid, methyl ester. The presence of phytochemicals with strong pharmacological actions like antimicrobial and antioxidants activities could be considered as sources of quality raw materials for food and pharmaceutical industries. This study further set a platform for isolating and understanding the characteristics of each compound for it pharmacological properties.

Biochemical and molecular characterization of cowpea landraces using seed storage proteins and SRAP marker patterns

Seven landraces of cowpea [Vigna unguiculata (L.) Walp.] were assessed for genetic variability in total proteins, protein fractions viz. albumins, globulins, prolamins, and glutelins by SDS-polyacrylamide gel electrophoresis and DNA polymorphism using sequence-related amplified polymorphisms (SRAP) markers. The solubility-based protein fractionation data indicated that the salt soluble fraction (globulin) and water-soluble fraction (albumin) proteins were the predominant fractions in cowpea seeds comprising 45–50.3% and 31.2–35.5% of total soluble proteins, respectively. The electrophoretic pattern revealed the molecular heterogeneity among total proteins as well as different protein fractions. The molecular weights of protein bands obtained by SDS-PAGE varied between 10 to 250, 15 to 110, 15 to 150, and 15 to 130 kDa for total proteins, albumins, globulins, and glutelins, respectively. A large number of bands were found common to the various landraces, indicative of their close relationship with one another. However, a few bands distinctive to some specific landraces were also detected, indicating varietal differences. A 34 SRAP primer pair combination generated a total of 1003 amplicons (loci) showed 100% polymorphism with an average of 0.93 polymorphism information content (PIC) value. Landraces displayed an average 0.50 similarity coefficient which clustered the landraces corresponding to their growth habit in main clusters and to their geographical origin in subcultures. Molecular and biochemical analysis were correlated with a medium level (Mantel test, r = 0.56, P < 0.02). These findings revealed that seed proteins and DNA polymorphism provide valuable information regarding the variability among landraces and this information could be utilized for breeding purposes in the enhancement of protein quality and quantity in grain legumes.

Morphological and Genetic Diversity of Cereal Genotypes in Kingdom of Saudi Arabia

Cereals are rich in carbohydrates and offer a major source of daily calorie intake. Among cereals, wheat (Triticum aestivum L.), barley (Hordeum vulgare L.), maize (Zea mays L.) and sorghum (Sorghum bicolor L.) are the leading crops grown across the globe. The utilization of exotic and diverse germplasm of a crop is a useful tool to increase the genetic diversity among the genotypes. In this study, we collected 11 wheat, 4 barley, 15 sorghum and 6 maize genotypes from farmer’s fields of Saudi Arabia and evaluated, for their morphological and genetic diversity potential. Significant differences were observed in the morphological characters of tested wheat, barley, sorghum and maize genotypes under field conditions. Sequence-related amplified polymorphism showed substantial genetic diversity in the tested genotypes of all the cereal crops. All the genotypes of wheat and barley significantly differed for the plant height, productive tillers, 1000 grain weight, and days to 50% flowering and maturity. Similarly, sorghum and maize genotypes differed significantly for the leaf area and plant height. All genotypes of wheat, barley, sorghum and maize differed for the number of alleles; maximum alleles were 156 in wheat, 172 in barley, 127 in sorghum and 73 in maize, per primer combination. Polymorphism of all of tested genotypes of wheat, barley, sorghum and maize was 100% in all the tested genotypes. Existence of genetic diversity of these tested wheat, barley, sorghum and maize genotypes offers opportunities to exploit favourable alleles for use in the breeding program aimed at yield improvement.