ISSR, RAPD and agronomic study in some F1 and F2 cotton genotypes

Cotton is an important economic crop plant with diploid and tetraploid cultivars. Hybridization is one of the main breeding strategies in cotton breeding producing new genotypes and also increasing the genetic diversity in cotton germplasm available. The present study considers agronomic and molecular study of genetic variations in thirteen F1 and F2 cotton genotypes (Gossypium hirsutum) obtained by crossing the cotton cultivars Bellizovar, No. 200, Siokra, Sindose and Tabladilla. Out of 30 RAPD primers used 19 primers produced 191 reproducible bands/loci out of which 63 bands were polymorph. Eight ISSR primers used produced 86 reproducible bands, out of which 27 bands were polymorph and 59 bands were monomorph. Some bands were present in the F1 progenies but absent in the F2 progenies of the same genotype. The mean values of gene diversity (H) and Shanon’s Information Indices (I) for ISSR markers in the F1 progenies were 0.27 and 0.40 respectively, while the same values in F2 progenies were 0.18 and 0.26 respectively. Similarly the mean values of H and I of the F1 progenies for RAPD markers were 0.07 and 0.11 respectively, while the same values in F2 progenies were 0.09 and 0.13 respectively. UPGMA and NJ dendrograms grouped the F1 and F2 progenies of Siokra X Belilzovar together, standing from the other genotypes due to their genetic differences. The use of present finding in planning future hybridization is discussed

Inter-population genetic diversity in Olea cuspidata subsp. cuspidata revealed by SSR and ISSR markers

The olive tree (O. europaea subsp. europaea var. europaea) is an ancient economic plant species with both cultivated and wild forms. In the present study, simple sequence repeat; SSR and inter simple sequence repeat; ISSR markers were used to study the genetic variation present in a few wild olive (O. europaea subsp. cuspidata) populations growing in the southern and western parts of Iran. Five SSR loci were analyzed which showed the presence of 12.6 alleles in average. The observed heterozygosity obtained for SSR markers ranged from 0.444 to 0.795 (mean value = 0.625), while Shannon index and polymorphic information content ranged from 1.46 (UDO99-043) to 2.47 (DCA3) and 0.620 (UDO99-43) to 0.871 (DCA3) respectively. Similarly ISSR analysis produced a total of 41 reproducible bands ranging from 240 bp (UBC-807) to 1750 bp (UBC823). The highest Nei’s genetic diversity as well as Shannon index was obtained for UBC823 locus (0.37 and 0.54 respectively). Kerman population had the highest mean number of alleles while Jareh population showed the lowest number of alleles. Analysis of molecular variation (AMOVA) showed significant difference (p < 0.05) both among and between olive populations for SSR supported by Fst pair-wise test. The highest value of within population ISSR genetic diversity occurred in Pahtak population, while Jareh population showed the highest value of among populations genetic diversity. UPGMA and NJ dendrograms obtained based on SSR and ISSR markers grouped the individuals of each population together in a distinct cluster, separated from the other populations due to their genetic distinctness

Study of intracultivar variation among main Iranian olive cultivars using SSR markers

Three Iranian olive cultivars of Geloleh, Shengeh and Rowghani with commercial interest are distributed in 3 provinces in North of Iran. Fifty one accessions belonging to these 3 olive cultivars were screened by 13 microsatellite markers revealing high genetic variability both within and between cultivars. In total, 54 alleles were detected with a mean number of 4.2 alleles per locus. Six unique allelic patterns were observed. Heterozygosity ranged from 0.00 to 1.00 while the mean number of polymorphic information content (PIC) was 0.51. The existence of homonyms, synonyms or mislabeling as well as intracultivar polymorphism was showed by allele differences between olive accessions studied. The phenogram obtained by UPGMA clustering showed variability among as well as between cultivars

Species relationship in the genus Silene L. Section Auriculatae (Caryophyllaceae) based on morphology and RAPD analyses

Morphological and RAPD studies were performed on Silene species of the sect. Auriculatae growing in Iran for the first time using phenetic, parsimony and Bayesian analyses. Trees obtained differed in the species groupings although agreed in some parts. Parsimony and Bayesian analyses of morphological characters produced some clades which were not well supported by bootstrap and clade credibility values but UPGMA tree showed a high cophenetic correlation. Grouping based on morphological characters partly support the species affinity given in Flora Iranica. Out of 40 RAPD primers used 15 primer produced reproducible polymorphic bands. In total 347 bands were produced out of which 340 bands were polymorph and 7 bands were monomorph. Among the species studied S. goniocaula showed the highest number of RAPD bands (184), while S. commelinifolia var. isophylla showed the lowest number (123). Some of the species studied showed the presence of specific bands which may be use for species discrimination. NJ and Bayesian trees of RAPD data partly agree with morphological trees obtained

Preliminarily report on molecular diversity of Sargassum species in Oman Sea by using ISSR and RAPD markers

Sargassum (Sargassaceae, Fucales) is a common macroalgal genus occurring throughout the world, except in the polar regions. Sargassum species are one of economically important brown algae in south of Iran. In this study, molecular variations were assessed in three Sargassum species; Sargassum tenerrimum J. Agardh, Sargassumglaucescens J. Agardh and Sargassum ilicifolium C.Agardh, widely distributed species in the southwest of Iran (Oman Sea). RAPD and ISSR markers were used to assess genetic variation within populations of each 3 species. Four of 30 RAPD primers as well as six combination of RAPD primers which have been used which all produced reproducible bands with high polymorphism (>96%). All populations in 3 species showed unique alleles which made unique profiles for each population. Twelve ISSR markers including single and combined primers showed high polymorphism (>94%). Nei’s genetic diversity, Shannon index showed high values between populations while no variations were observed within populations (Hpop =0, 1-Hpop/Hsp =1) in both molecular markers studied. AMOVA test also confirmed lack of variation within them. Different clustering like UPGMA and Neighbor Joining separated populations of each species studied based on RAPD and ISSR data. This is the first study on evaluation of inter-population variation in some of Sargassum species in Iran

Inter-populations genetic and morphological diversity in three Silene (Caryophyllaceae) species

The inter-populations morphological and genetic variations were studied in three species of Silene (Silene indeprensa, Silene gynodioica and Silene crispans) of the section Auriculatae, which grow and form several populations in different regions of Iran. Morphological analysis of variance (ANOVA) and analysis of molecular (AMOVA) analyses showed the species distinctness. Unweighted paired group with arithmetic average (UPGMA) clustering of morphological characters and neighbor-joining (NJ) tree of molecular features almost separated the species from each other. In S. gynodioica, Soltaneiyes and Gheydar populations differed from the other populations in both morphological and molecular features, and in case of S. indeprensa, Ghorkhood and Hezarmasjed populations differed significantly from the other populations in both morphological and molecular features. Therefore, these populations are considered as a new subspecies in these two species. Some of the populations showed the presence of specific inter-simple sequence repeat (ISSR) band/locus, indicting genetic divergence of the populations possibly either due to local adaptations or genetic drift. Mantel test performed did not show significant correlation between morphological/molecular distance and geographical distance of the populations studied.Keywords: Diversity, ISSR, morphometry, Silen

Molecular and genome size analyses of somaclonal variation in apple rootstocks Malling 7 and Malling 9

The cultivated apple (Malus domestica) is important fruit crops cultivated in world. For production and breeding of high quality apple, inducing and enhancing new genetic diversity and suitable traits are necessary. In Iran, different local and imported apple genotypes are cultivated and common apple root stocks (Malling 7) M7 and (Malling 9) M9 are mostly used root stocks in the country. Therefore, we studied genetic diversity of M7 an M9 tissue culture regenerated plants produced by different treatments by using twenty ISSR markers. In total 51 randomly selected plants were studied for the occurrence of somaclonal variation in apple mother plants and tissue culture regenerated plants. Four different treatments were used for tissue culture. Genetic diversity parameters, genetic distance and polymorphism percentage were studied in regenerated plants. Variations in loci frequency and combination were checked by STRUCTURE and the presence of similar loci in the plants was studied by reticulation NJ tree. Genetic relationship versus distinctness was determined by principal coordinate analysis. The results showed the occurrence of genetic variation among mother plants and tissue culture regenerated plants of each subculture due to somaclonal variation. Significant difference in the genome size among some of the regenerated plants indicates that change in genetic structure of plants during tissue culture is also accompanied with quantitative change in DNA. However, degree of genetic variation differed among apple rootstocks and also among different treatments used

Species delimitation and genetic diversity analysis in Salvia with the use of ISSR molecular markers

Thirty-nine plant specimens of six Salvia species were collected from different localities of the Alborz mountain region in Iran and studied for morphological and genetic variability and species relationship. Inter simple sequence repeats (ISSR) molecular markers showed a high degree of within-species and interspecific genetic variability in Salvia. Analysis of molecular variance and Hickory tests showed significant molecular difference among the studied populations. A principal coordinate analysis plot of morphological characters grouped the species into two distinct groups, supporting their taxonomic treatment. This was partly supported by ISSR networking. The Mantel test did not show a correlation between genetic distance and the geographical distance of the studied populations. STRUCTURE and reticulation analyses revealed some degree of gene flow among the species. The present study showed that ISSR molecular markers could be used in Salvia species delimitation along with morphological study

Assessment of Relationships Among and Within Helichrysum Mill. (Asteraceae) Species by Using ISSR Markers and Morphological Traits

AbstractThis study conducted to determine relationship among and within Iranian Helichrysum species (Asteraceae), using molecular analyses which are based on the inter simple sequences repeat (ISSR) technique and morphological observations of taxa which are collected in the areas of different habitats of Iran, where 20 species have been recorded within this genus. Helichrysum is a large and taxonomically difficult genus and relationships and infrageneric classification have remained largely unresolved. In this study Based on ISSR markers, ten primers generated 1145 polymorphic ISSR-PCR bands. Cluster analysis by the unweighted pair-group method (UPGMA) separated the 64 populations into two main clusters that were in agreement with the principal coordinate (PCO) analysis. Both UPGMA and the PCoA demonstrated that the highest percentage of ISSR loci polymorphism (54.7 %) occurred in populations of H. armenium. The highest gene diversity over loci (1.224), Shannon's Information Index (0.224 %) and Expected Heterozygosity (0.142 %) occurred in H. armenium (0.18) and the lowest of these parameters (0%) were observed in H. araxinum, H. graveolens, H. persicum and H. psychrophayllum. The highest genetic similarity occurred between H. armenium and H. rubicundum (0.989), while the lowest was between H.polyphyllum and H. graveolance (0.213). The analysis of molecular variance (AMOVA), showed significant genetic variation among (24%) and within (76%) species. A morphological analysis of this genus is carried out using 36 quantitative and qualitative morphological traits in 87 populations (249 individuals) of 19 species and traits such as indumentum, resting bud, achene length, achenial papillae, dimension of receptacle and form and apex of phyllaries were main diagnostic features. A high degree of similarity was revealed between H.artemisioides and H. davisianum.Results obtained from the morphological cluster were greatly consistent with the molecular data, to elucidating taxonomic relationships, as well as both attributed the higher diversity in H. armenium and H.rubicundum in comparison with other species and also indicated that H. persicum is a member of H. oocephalum species. Totally we confirmed the presence of 18 species in Iran

Genome size, morphological and palynological variations, and heterostyly in some species of the genus Linum L. (Linaceae) in Iran

Heterostyly is the occurrence of flowers with different sexual organ arrangements in different plants of the same species. This floral polymorphism occurs in four sections of genus Linum. The present study compares the morphological, palynologycal and genome size (C-value content) characteristics in the long-styled and short-styled plants in three Linum species, that is, Linum austriacum L., Linum album Ky.ex Boiss. and L. glaucum Boiss. &amp; Nöe . 15 qualitative and quantitative morphological characters from both vegetative and reproductive organs of these plants were studied. A higher mean value of the plant height, size of the basal leaves width, flower leaves width, calyx length, sepal length and petal length occurred in the long-styled plants, while the mean value of branch number, basal leaves length, flower leaves length, calyx width, pedicel length and sepal length was higher in the short-styled plant populations. T-test analysis of morphological characters showed significant difference (p&lt;0.05) for some of the characters studied. Principal coordinate analysis (PCoA) plot of long-styled and short-styled plant populations based on all morphological characters also separated these two kinds of plants in the three species studied. The pollen obtained from the mature buds was prepared for light microscopy (LM) and scanning electron microscopy (SEM). The polar and equatorial views of the pollen grains were similar in the long-styled (Ls) and short-styled (Ss) plants but the aperture shape differed in these populations. C-values obtained by flow cytometry, differed in the long-styled and short- styled plants of the species studied. The analysis of variance (ANOVA) test performed among the three Linum species showed a significant difference in 2C-value content. Positive significant correlation was observed between 2C-value and northern distribution of the Linum species studied, while a negative significant correlation occurred with eastern distribution.Key words: Heterostyly, Linum, morphology, palynology, genome size