Random Amplified Polymorphic DNA Data Research Articles

Genetic diversity in Corchorus olitorius L. revealed by morphophysiological and molecular analyses.

Assessment of genetic diversity has an efficient role in plant breeding and improvement programs. There is a limit number of investigations dealing with the evaluation of genetic diversity in Jew's mallow (Corchorus olitorius L.), despite its valuable importance as a leafy vegetable and a delicious dish rich in vitamins and antioxidants. Therefore, in this study, 18 landraces of Jew's mallow-collected from different locations in Egypt-were used for genetic diversity assessment based on morphophysiological and molecular evaluations. A high degree of variability was found among the evaluated landraces at both levels, indicating the appropriateness of such collection to be involved in breeding approaches. Some morphophysiological traits offered a high level of diversity and effectively discriminated the landraces. Thus, they are recommended to be used in successive morphological evaluation studies. On the other hand, molecular evaluation using the random amplified polymorphic DNA (RAPD) and the sequence related amplified polymorphism (SRAP) efficiently supported the morphological results by exposing a clear genetic relationship among the landraces. In addition, the principal coordinate analysis based on combined data of RAPD and SRAP divided the landraces into two main groups, reflecting their relationship molecularly. The first group included nine landraces related to Upper Egypt and the second gathered three landraces from Delta, while the other six landraces were distinctly distributed around these two groups. The two groups may have two distinct ancestors in addition to the different ancestors of the scattered landraces. Findings of this study are valuable and could assist in Jew's mallow breeding programs.

GENETIC VARIATION OF WILD Musa acuminata COLLA FROM INDONESIA

Indonesia is the center of origin and diversity of Musa acuminata Colla, one of the progenitors of cultivated bananas today. However, the genetic variation of wild M. acuminata has not been studied extensively, thus this study aimed to assess the genetic variation of the Indonesian wild M. acuminata based on 10 RAPD (Random Amplified Polymorphic DNA) and 10 ISSR (Inter Simple Sequence Repeats) markers. The genetic properties of 17 populations of wild M. acuminata were analyzed by Popgene 1.32 based on Nei’s unbiased measures of genetic identity and genetic distance. Of the 443 DNA bands produced, 425 (95.94%) were polymorphic. Cluster analysis of the combined data of RAPD and ISSR produced a dendrogram which separated the population of M. acuminata (A genome) from M. balbisiana (B genome), but not from M. schizocarpa (S genome). Nei’s genetic distance of the 17 populations of wild M. acuminata ranged from 0.3676 to 0.1634. The highest genetic distance was observed between M. acuminata var rutilifes (from East Java) and M. acuminata var sumatrana (from West Sumatra). The percentages of polymorphic loci among the 17 populations of M. acuminata ranged from 9.93% to 39.73%. Nei’s gene diversity (h) ranged from 0.041 to 0.1418. M. acuminata var malaccensis population was the most diverse among the researched 17 M. acuminata. The high level of genetic diversity of the wild M. acuminata from Indonesia emphasizes the need for conservation and preservation of the natural population and its use in the banana breeding program.

Random amplified polymorphic DNA-based molecular heterogeneity analysis of Salmonella enterica isolates from foods of animal origin.

Aim:This study aims to study the significance of random amplified polymorphic DNA (RAPD) typing in heterogeneity analysis of Salmonella serovars, isolated from foods of animal origin.Materials and Methods:Salmonella serovars isolated and identified from different foods of animal origin such as meat, milk, and egg by standard bacteriological methods. DNA isolated from all 10 isolates which are confirmed by biochemical and serotyping methods and then RAPD was performed using the primers OPB 10, primer 1290, NSC I, NSC II, and primer 3. Then, RAPD data were analyzed using the BioNumerics software, Belgium, Germany.Results:RAPD polymerase chain reaction (PCR) using five primers, namely OPB 10, primer 1290, NSC I, NSC II, and primer 3, classified the 10 isolates into 9, 10, 10, 7, and 10 RAPD-PCR types with discriminating powers of 0.1987, 0.423, 0.50889, 0.1842, and 0.2582, respectively. The phylogram constructed with NSC I profile classified isolates based on geographical origin. Primer 1290, NSC II, and primer 3 produced some uniform bands in all isolates indicating their binding ability in conserved genomic region. This study revealed that RAPD profile can be best used for finding out the heterogeneity at molecular level of Salmonella isolates in combination with other molecular and phenotypic typing techniques. Thus, our results support earlier observation of its significance by different workers on different Salmonella serotypes.Conclusion:Repeatability of RAPD-PCR is insufficient to distinguish genetic differences among Salmonella serovars.

Genetic Diversity among Geographically Separated <i>Cyperus rotundus</i> Accessions Based on RAPD Markers and Morphological Characteristics

The diversity of globally distributed populations of purple nutsedge was assessed using molecular marker data and morphological traits. Cluster analysis of binary random amplified polymorphic DNA (RAPD) data and morphological traits indicated that the global population of purple nutsedge consisted of two clades. Cluster analysis of the RAPD data supported separation of the purple nutsedge accessions evaluated into two distinct clades of 11 and 33 accessions. Except for accessions identified as California* and Arizona, all USA accessions were clustered with accessions from Taiwan, Western Samoa, New Zealand, Malaysia, Japan, El Salvador, Columbia, Australia, Thailand and West Indies. A second cluster included accessions from Sudan, Greece, Iran, California*, Arizona, Brazil, Argentina, Mauritius, Philippines, Indonesia and Tanzania. The accessions from Sudan, Greece, Iran, Mauritius, and Tanzania were distributed along a similar longitudinal axis. Cluster analysis based on morphological traits though not identical to that based on RAPD data also supported separation into two clades and perhaps a third. The lack of genetic diversity among accessions supported the hypothesis that spread and propagation into new environments were largely by tubers which preserved genetic identity. The lack of diversity particularly among New World and USA accessions may also reflect a relatively recent introduction of the species into the Americas and a low level of outcrossing.

Assessment of Genetic Diversity and Evaluation of Relatedness Through Morphological and Molecular Markers Among Medicinally Important Trees: Terminalia arjuna, T. bellerica, T. catappa and T. chebula

Terminalia trees are being over exploited because of their medicinal and economical importance leading to loss of valuable genetic resources. The study included morphological variation and characterization of genetic diversity at interspecific level. Genus Terminalia is a complex group having many problems associated with taxonomic identification. Morphologically T. arjuna can be distinguished from other species having winged fruits whereas the fruits of T. catappa are elliptical and tomentose. Fruits of T. chebula are provided with ribs while fruits of T. bellerica are without ribs. Thus different species of Terminalia can be distinguished by characteristic of fruits and sometimes by characteristic of leaves. For the first time comprehensive study was done for estimation and analysis of genetic variation through random amplified polymorphic DNA (RAPD) and inter-simple sequence repeats (ISSR) markers. A total of 12 polymorphic primers (6 RAPD and 6 ISSR) produced 80 polymorphic bands with an average of 7.5 bands per primer. The average polymorphism was 87.97% and five primers were reported to produce 100% polymorphism. Dendrogram based on RAPD and ISSR data grouped four Terminalia species in two distinct clusters. Cluster-I comprised 2 species viz. T. arjuna, T. chebula, while cluster-II comprised two species T. bellerica and T. catappa. Clustering of species was altercating with previous morphological studies.

Isolation and molecular identification of Lactobacillus with probiotic potential from abomasums driven rennet

Lactobacillus species are beneficial for the functional food industry and preventive medicine. The complex microflora of traditional cheese depends on the cheese types (e.g., homemade rennets). Here, the abomasum driven rennet was assessed for the existence of lactobacilli. For differentiating lactobacilli, the bacterial suspension was screened for the acid and bile resistance. The isolated bacteria were evaluated for antibiotic susceptibility and antagonistic impacts on other pathogenic bacteria. The 16S rDNA gene was evaluated by the amplified ribosomal DNA restriction analysis (ARDRA) recruiting the restriction enzyme Taq I and compared to the virtually digested patterns of previous reports on lactobacilli. The isolates were examined by random amplified polymorphic DNA (RAPD) and distinctive lactobacilli were sequenced. ARDRA and RAPD data showed three distinct lactobacilli strains, including L. acidophilus, L. planetarum, and L. fermentum. The homemade rennet is proposed as the novel source of probiotic strains as an alternative to the traditional cheeses.

Genetic Fingerprinting of Two Species of Averrhoa Using RAPD and SRAP Markers

Random amplified polymorphic DNA (RAPD) and sequence-related amplified polymorphism (SRAP) marker analyses were conducted in two underutilized fruit trees of the family, Oxalidaceae, namely Averrhoa bilimbi and A. carambola, commonly known as cucumber tree and starfruit tree respectively. The current study focused on the utility of 23 RAPD primers and 31 SRAP primers. The RAPD primer, OPD-07 produced the greater number of amplified fragments, exhibited higher level of polymorphism and could differentiate between almost all the collections on the basis of the efficiency of different primer parameters. The most competent SRAP primer combination for the characterization of Averrhoa collections was ME2/EM6. Cluster and PCA analyses on both RAPD and SRAP data differentiated the A. bilimbi and A. carambola collections. Both markers showed that A. bilimbi collections, Ab2, Ab3 and Ab4 were distinct from the rest. Both SRAP and integrated data were capable of distinguishing sour and sweet collections of A. carambola. The present study provides the first in-depth report on the molecular diversity between the two species of the genus Averrhoa with regard to RAPD and SRAP markers. Both RAPD and SRAP could discriminate the species, and collections of Averrhoa. SRAP are the preferred choice because it differentiates various collections and possess good amplification, stability and polymorphism.

Genetic and ecogeographical diversity of rye (Secale L.) species growing in Iran, based on morphological traits and RAPD markers

Assessing the genetic diversity is an essential step for breeding programs and the optimal use of genetic resources. In this study genetic and phenotypic diversity of 70 rye (Secale L.) populations collected from different regions of Iran were evaluated based on 14 qualitative morphological traits and Random Amplified Polymorphic DNA (RAPD) markers. Among the evaluated traits spike shattering, leaf pubescence and stem and auricle color showed a high taxonomic value. Based on the morphological diversity assessed using descriptive statistics and discriminant function analysis, 38 S. cereale L. subsp. cereale, 28 S. cereale L. subsp. ancestrale Zhuk., eight S. strictum C. Presl, subsp. strictum, and three populations of S. strictum C. Presl, var. ciliatoglume (Boiss.) Fred. et Peters. were identified. Genetic diversity of 20 populations of identified species by RAPD markers confirmed the results of morphological evaluation. The rate of polymorphism was 45.26% of total amplified DNA fragments. Cluster analysis of RAPD data effectively separated the two species of rye (S. cereale L. and S. strictum C. Presl).

Molecular characterization of diversity in Annona squamosa L.: A high value tropical medicinal plant

Introduction: Annona squamosa L. commonly known as custard apple, is an important under-utilized fruit species from tropical and subtropical parts of India with high potential as commercial horticultural crop. The aim of this study was to determine the genetic diversity among populations spread across districts of Udaipur, Rajsamand and Chittaurgarh falling within Aravallis in Rajasthan. In all, 21 populations were identified and subjected to molecular evaluation using random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) markers. The cumulative analysis carried out on the basis of RAPD and ISSR data sets revealed 73.91% polymorphism. Methods: A total of 251 amplicons were produced using 19 RAPD and 15 ISSR primers. The pair-wise distance matrix calculated by Dice’s co-efficient showed a distance range of 0.67 to 0.95 when computed using cumulative data set. Results: Characterization of genetic diversity within populations based on combined data set showed that percentage of polymorphism bands (PPB) ranged from 29.19 to 50.93. Nei’s gene diversity (HE) values varied from 0.1002 to 0.1964 within a total genetic diversity (HT) of 0.2509. The average observed number of alleles per locus (Na) was 1.3602 and Ne was 1.2400. The Shanon’s index was 0.2002 (Hpop) and 0.3800 (Hsp) at population and species level, respectively. Conclusion: The moderate gene flow value (Nm=0.59) and significant coefficient of genetic differentiation (GST=0.458) revealed low level of genetic diversity within population and highly significant genetic differentiation among populations of A. squamosa. The UPGMA dendrogram resolved all the 21 populations representing 3 districts into various clusters according to their genetic distances. The principal component analysis (PCA) supported the UPGMA results and justified the relationship among populations.

Estimating Genetic Conformism of Korean Mulberry Cultivars Using Random Amplified Polymorphic DNA and Inter-Simple Sequence Repeat Profiling

Apart from being fed to silkworms in sericulture, the ecologically important Mulberry plant has been used for traditional medicine in Asian countries as well as in manufacturing wine, food, and beverages. Germplasm analysis among Mulberry cultivars originating from South Korea is crucial in the plant breeding program for cultivar development. Hence, the genetic deviations and relations among 8 Morus alba plants, and one Morus lhou plant, of different cultivars collected from South Korea were investigated using 10 random amplified polymorphic DNA (RAPD) and 10 inter-simple sequence repeat (ISSR) markers in the present study. The ISSR markers exhibited a higher polymorphism (63.42%) among mulberry genotypes in comparison to RAPD markers. Furthermore, the similarity coefficient was estimated for both markers and found to be varying between 0.183 and 0.814 for combined pooled data of ISSR and RAPD. The phenogram drawn using the UPGMA cluster method based on combined pooled data of RAPD and ISSR markers divided the nine mulberry genotypes into two divergent major groups and the two individual independent accessions. The distant relationship between Dae-Saug (SM1) and SangchonJo Sang Saeng (SM5) offers a possibility of utilizing them in mulberry cultivar improvement of Morus species of South Korea.

Assessment of RAPD Markers to Analyse the Genetic Diversity among Sunflower (Helianthus annuus L.) Genotypes

Genetic diversity estimation among different species is an important tool for genetic improvement to maximize the yield, desirable quality, wider adaptation, pest and insect resistance that ultimately boosting traditional plant breeding methods. The most efficient way of diversity estimation is application of molecular markers. In this study, twenty random amplified polymorphic DNA (RAPD) primers were utilized to estimate the genetic diversity between ten sunflower genotypes. Overall 227 bands were amplified by 20 primers with an average of 11.35 bands per primer. RAPD data showed 86.34% polymorophic bands and 13.65% of monomorophic bands. Genetic similarity was ranged from 50.22% to 87.22%. The lowest similarity (50.22%) was observed between FH-352 and FH-359 and the maximum similarity 87.22% was observed between A-23 and G-46. Polymorphic information content (PIC) values were varying from 0.05 to 0.12 with a mean of 0.09. Cluster analysis based on RAPD results displayed two major distinct groups 1 and 2. Group-2 contains FH-352 which was the most diverse genotype, while group-1 consists of few sub groups with all other genotypes. Ample diversity was found in all the genotypes. Present study reveals novel information about sunflower genome which can be used in future studies for sunflower improvement.

Variabilities on DNA Level of Two Selected Potato Genotypes under Seawater Salinity

Random Amplified Polymorphic DNA (RAPD) banding patterns of selected potato (Cara) and (Lady Rosetta), under seawater salinity stress, on agarose gel showed (9 and 10) distinct RAPD profiles generated by five of different 10-mers oligonucleotides primers. The recorded RAPD data of both potato genotypes showed highly polymorphism and low levels of similarity between and inside the treated samples and the untreated control sample. The RAPD analysis of the treated plants of Lady Rosetta showed 363 amplified bands of the treated and control plants whereas the treated plants distributed as ~50.41% unique polymorphic bands, ~29.75% of repeated polymorphic bands and ~6.89% of monomorphic bands as compared with the control plants which showed ~8% unique polymorphic bands and ~4.95% monomorphic bands. However, with Cara, the RAPD analysis showed 227 amplified bands of the treated and control plants whereas the treated plants distributed as ~48.02% of unique polymorphic bands and ~31.28% of repeated polymorphic bands and ~4.4% of monomorphic bands as compared with the control plants which showed ~13.22% unique polymorphic bands and ~3.08% monomorphic bands as compared with the treated plants. There is no doubt that, the gain or the loss of parts of the DNA may consider as speculation the reason for salinity tolerance in the presented work. On the other hand, the exposure to the salinity stress leads to induce changes on the DNA level.

Molecular and phytochemical analysis of wild type and olive cultivars grown under Saudi Arabian environment.

This study aimed to assess genetic variability at molecular and phytochemical levels among the four most commonly grown olive cultivars and the wild-type olive of Saudi Arabia. Sixty-six and 80 amplicons were generated from 9 random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) primers, each, producing an average of 95.9 and 86.44% polymorphism for the two markers, respectively. The PIC values were 82.2% for the RAPD and 85.4% for the ISSR markers and the discrimination power for both the markers was 11.1%. The UPGMA cluster analysis based on the RAPD and ISSR data resulted in the aggregation of cultivars and wild accession with a good bootstrapping value according to their origin. Furthermore, a total of 199 compounds were identified in the cultivars based on peak area, retention time, and molecular formula using GC-MS analyses of methanolic and ethanolic extracts. These compounds were classified according to their chemical class; most of them were fatty acids, alcoholic compounds, carboxylic acids, aldehydes, heterocyclic compounds, ketones, alkanes, and phenols. Genetic and phytochemical distances were significantly correlated, based on the Mantel test. The Saudi wild accession also had high numbers of fatty acids and their esters, and can be used in breeding programs for generating new genotypes with interesting characters.

Genetic Diversity Assessment of a Critically Endangered Medicinal Plant Populations from Shorea tumbuggaia Roxb.

A successful assessment of genetic diversity is consider vital for formulating conservation of the rare, endangered and threatened (RET) species. Our aim was to assess the genetic diversity in Shorea tumbuggaia Roxb. population (Dipterocarpaceae family)—a critically endangered medicinal plant of conservative interest using Random Amplified Polymorphic DNA (RAPD) technique. According to International Union for Conservation of Nature (IUCN) and Conservation Assessment and Management Planning (CAMP) S. tumbuggaia is considered as an endemic and globally endangered tree species. RAPD markers were used to estimate the genetic variability and similarity among the S. tumbuggaia. Eighteen (18) primers of OPU (1-20) series generated a total of 137 polymorphic bands out of 735 total bands (19.86% polymorphism), with an average of 40.83 amplified bands per primers and 598 bands showed monomorphic banding pattern with an average of 33.22 per primers. Genetic similarity coefficients calculated from RAPD data ranged from 0.82 to 0.95, with the highest value of 0.95 in ST10 and ST11 and the lowest value of 0.82 between accessions ST2 and ST9, ST3 and ST4, ST4 and ST10 and ST5 and ST8.

Genetic Diversity and Identification of Variety-specific RAPD and ISSR Markers in Foeniculum vulgare

Inter simple sequence repeats (ISSRs) and random amplification of polymorphic DNA (RAPD) were used to determine genetic relationships and variety specific markers in fennel varieties grown in Rajasthan. In this study, the percentage polymorphism revealed by RAPD and ISSR was 50 and 39.1%, respectively. Dendrogram constructed from unweighted pair group method with arithmetic mean cluster analysis based on RAPD and ISSR data showed clear genetic relationships among these germplasms. Similar clustering was obtained when another software DARwin 5 was used to construct tree plot from factorial analysis based on RAPD and ISSR data. Besides, from the bands amplified, there were three independent ways to identify the fennel varieties, such as unique RAPD and ISSR markers, unique band patterns and a combination of the band patterns provided by different primers. Finally, the molecular fingerprints have been generated for five fennel varieties, namely FNL-41, 46, RF-125, 143 and 178. These could be distinguished by using specific 10 RAPD and four ISSR markers, which showed either presence or absence of bands generated by seven RAPD and four ISSR primers.

Diallel analysis and genetic diversity of some yellow maize inbred lines (Zea mays L.) using RAPD and SSR markers

Information about combining ability and genetic diversity among maize inbred lines is fundamental in designing breeding strategies for improving grain yield. Twenty one F1 hybrids were generated by crossing seven yellow maize inbred lines in a half diallel mating scheme in 2013 season. The 21 F1 hybrids plus the two check hybrids (SC166 and SC173) were evaluated in a randomized complete block design with three replications during 2014 season, to estimate combining ability effects and identify type of gene action governing the inheritance of grain yield and other important agronomic traits. Results showed that both general (GCA) and specific (SCA) combining ability mean squares were highly significant for all the studied characters. The GCA/SCA ratio was more than unity for all the studied traits, except days to 50% silking, plant height and grain yield, indicating the preponderance of the additive gene effects in the inheritance of these traits. The inbred line P1 appeared to be the best general combiner for earliness, grain yield and its components. The crosses P1×P2, P3×P7, P4×P7, and P5×P6 had the best SCA effects for grain yield and most of its component traits. The cross P1×P2 significantly out-yielded the two check hybrids (SC166 and SC173). The genetic diversity among the seven yellow parental inbred lines was assessed using two types of molecular markers; Random Amplified Polymorphic DNA (RAPD) and Simple Sequence Repeats (SSR). Seven random primers were used in the RAPD reactions, resulting in the amplification of 82 bands with 95.12% polymorphism. While, eight pairs of SSR primers were used and resulted in 49 fragments with 100% polymorphism. Genetic similarity among all possible pairs of inbred lines varied from 0.12 to 0.69, with an average of 0.34, for RAPD markers, and from 0.01 to 0.56, with an average of 0.24, for SSR markers. The similarity matrices for RAPD and SSR data were not significantly correlated (r=0.34, p >0.05). Genetic distances based on RAPD and SSR markers were insignificantly correlated with F1 hybrids grain yield, the correlation values being (r =0.02 and 0.429 P>0.05), respectively. Therefore, the parental genetic distance could not be used to predict the grain yield of the F1 hybrids in this study.

Genetic diversity, mating types and phylogenetic analysis of Indian races of Fusarium oxysporum f. sp. ciceris from chickpea

The present study describes the comparative analysis of five genetic markers viz., random amplified polymorphic DNA (RAPD), enterobacterial repetitive intergenic consensus (ERIC), BOX-elements, mating type (MAT) locus and microsatellites for genetic analysis of virulent isolates of Fusarium oxysporum f. sp. ciceri (FOC) representing seven races from chickpea. Phylogenetic analysis of translation elongation factor 1-α and internal transcribed spacer region separated all the FOC isolates into two major clades. Majority of the isolates (FOC 63, FOC 33, FOC 40, FOC 100, FOC 6, FOC 22, FOC 31, FOC 79 and NDFOC 98) representing race 1, 2, 5 and 6 grouped in clade I, while isolates (FOC 90, FOC 108 and FOC 88) belonging to race 3, 4 and 7 were clustered in clade II. Isolates (FOC 33, FOC 40, FOC 17 and FOC 100) representing race 2 had MAT-2 loci, while race 1 isolates (FOC 63, FOC 72 and FOC 76) contained MAT-1 loci only. The principal component analysis (PCA) of RAPD, ERIC, BOX and microsatellite marker data explained 39.94, 39.98, 42.04 and 62.59% of the total variation among test isolates, respectively. Furthermore, there was no correlation existed between genetic diversity, virulence, race compositions or geographic origin of the isolates. Overall, these findings will assist in better understanding of the genetic variability and ideally, will improve disease management practices.

Correlation between the Chemical and Genetic Relationships among Thymus saturejoides Genotypes Cultured under in vitro and in vivo Environments.

In this study, the in vitro and in vivo essential oil (EO) composition and genetic variability in six micropropagated genotypes of Thymus saturejoides Coss., a Mediterranean medicinal and aromatic plant, were analyzed by GC/MS and randomly amplified polymorphic DNA (RAPD). Yield and composition of the EO varied between genotypes. Cluster analysis based on RAPD data and EO grouped the six genotypes in three groups in both culture conditions, thus showing considerable intraspecific genetic and chemical variations. Applying the Mantel test, the result showed a significant correlation between the two proximity matrices RAPD and EO obtained from in vitro genotypes, whereas this correlation was not observed when using the EO obtained from the in vivo genotypes.

Genetic Diversity in Haploid<i> Nicotiana alata</i> Induced by Gamma Irradiation, Salt Tolerance and Detection of These Differences by RAPD

Haploid plants of Nicotiana alata were cultured in vitro on MS medium with IAA + KIN. The resulting plantlets were irradiated using gamma radiation doses of 10, 15, 20 and 25 Gy. Single node pieces were cut and transferred onto fresh MS medium. Gamma radiation doses caused the death of 9% and up to 28% of explants. NaCl concentrations caused the death of 8% up to 36% of explants, while the combined effect between gamma radiation doses and salinity had an impact suffused on the percentage of survival. The combined effect of gamma radiation doses 20 Gy and 25 Gy on NaCl concentrations of 100, 150 and 200 mM were deadly. Even more, the combined effect of gamma radiation doses and salinity had a severe negative impact on both the proline content and total soluble protein. Random amplified polymorphic DNA (RAPD) analysis was used to determine the degree of genetic variation in treated haploid Nicotiana alata plants. Total genomic DNAs from different haploid plantlets treated were amplified using five arbitrary primers. Two hundred and seventy bands were detected from plantlets irradiated with doses of 15, 20 and 25 Gy, with polymorphic band number 226 (83.7%). The total number of bands resulted from plant grew on 150 mM and 200 mM NaCl were 260 bands with polymorphic bands 185 (85.6%). However, the total number of bands produced from combined effects between gamma rays and salinity (20 Gy X 50 mM NaCl, 20 Gy X 100 mM NaCl and 25 Gy X 50 mM NaCl) were 270, with polymorphic band number 231 (85.5%). High similarity between treatments was revealed. Treatments relationships were estimated through cluster analysis (UPGMA) based on RAPD data.

Genetic Diversity in Haploid<i> Nicotiana alata</i> Induced by Gamma Irradiation, Salt Tolerance and Detection of These Differences by RAPD

Haploid plants of Nicotiana alata were cultured in vitro on MS medium with IAA + KIN. The resulting plantlets were irradiated using gamma radiation doses of 10, 15, 20 and 25 Gy. Single node pieces were cut and transferred onto fresh MS medium. Gamma radiation doses caused the death of 9% and up to 28% of explants. NaCl concentrations caused the death of 8% up to 36% of explants, while the combined effect between gamma radiation doses and salinity had an impact suffused on the percentage of survival. The combined effect of gamma radiation doses 20 Gy and 25 Gy on NaCl concentrations of 100, 150 and 200 mM were deadly. Even more, the combined effect of gamma radiation doses and salinity had a severe negative impact on both the proline content and total soluble protein. Random amplified polymorphic DNA (RAPD) analysis was used to determine the degree of genetic variation in treated haploid Nicotiana alata plants. Total genomic DNAs from different haploid plantlets treated were amplified using five arbitrary primers. Two hundred and seventy bands were detected from plantlets irradiated with doses of 15, 20 and 25 Gy, with polymorphic band number 226 (83.7%). The total number of bands resulted from plant grew on 150 mM and 200 mM NaCl were 260 bands with polymorphic bands 185 (85.6%). However, the total number of bands produced from combined effects between gamma rays and salinity (20 Gy X 50 mM NaCl, 20 Gy X 100 mM NaCl and 25 Gy X 50 mM NaCl) were 270, with polymorphic band number 231 (85.5%). High similarity between treatments was revealed. Treatments relationships were estimated through cluster analysis (UPGMA) based on RAPD data.