Random Amplified Polymorphic DNA Primers Research Articles

Variability among selective guava (Psidium guajava L.) varieties revealed by morphology and RAPD marker

The research was conducted for the assessment of genetic diversity using both morphological and random amplified polymorphic DNA (RAPD) analysis of twelve guava (Psidium guajava L.) varieties growing in Bangladesh. Morphological characterization of guava varieties showed a wide range of variation. The highest variability was observed between Poly and Jelly varieties.Polymerase chain reaction with 5 arbitrary 10-mer and 3 arbitrary 12- mer RAPD primers produced a total of 50 bands of which 75.23 percent were polymorphic. The highest percentage of polymorphic loci (100%) was observed for primer A and the lowest (50%) for A03 primer. The UPGMA dendrogram revealed the segregation pattern and the difference of evolutionary changes. Guava varieties were separated into two main groups, one of them was made up of Chineese, Jelly, Kazi, Apple, L-49, Local-2 and Local-3. The other one was made up of Local-1, Poly, Kashi, Thai and Bombay. The highest genetic distance between Apple and Kazi peyara indicate that these varieties might be interesting in breeding programme for improving trait of interest. This scientific information could be used for further improvement of guava.
 Jahangirnagar University J. Biol. Sci. 7(2): 89-98, 2018 (December)

HUBUNGAN KEKERABATAN Shorea gysbertsiana DENGAN TIGA JENIS SHOREA PENGHASIL TENGKAWANG LAINNYA BERDASARKAN PENANDA RAPD

The Shorea tengkawang species; Shorea pinanga, S. stenoptera, S. macrophylla and S. gysbertsiana known for their high-value wood and non-wood products. Shorea tengkawang produces tengkawang oil which usually used as a raw material and supporting material for the manufacture of food, pharmaceuticals and cosmetics. This study determines the genetic relationship of S. gysbertsiana to another three Shorea species uses RAPD (Random Amplified Polymorphism DNA) specific loci. There were 13 specific alleles obtained from 6 RAPD primers. Among these specific alleles, there were 8 loci shared between S. gysbertsiana and S. macrophylla, and 2 loci shared with S. stenoptera. There were no specific loci shared between S. gysbertsiana and S. pinanga. This study found one specific locus for S. gysbertsiana and one specific locus between S. stenoptera and S. pinanga. These results revealed a very close genetik relationship of S. gysbertsiana to S. macrophylla and S. stenoptera.The specific loci found in this study can be used to support the morphological identification, also for supporting conservation program of these four Shorea species.

Photosynthetic response and DNA mutation of tropical, temperate and polar Chlorella under short-term UVR stress

Changes in photosynthetic efficiency of Photosystem II can be used as an early stress indicator in phototrophs. In this study, chlorophyll fluorescence, measured by a Pulse-Amplitude Modulated Fluorometer (PAM), was used to determine the photosynthetic performance of tropical, temperate, Antarctic and Arctic Chlorella in response to short-term acute ultraviolet radiation (UVR) stress, with measurements of maximum quantum yield (Fv/Fm), photosynthetic efficiency (α), maximum electron transport rate (rETRm) and photoadaptive index (Ek). Three light treatments were conducted over a continuous, five-hour duration: (i) control subjected only to photosynthetic active radiation (PAR), (ii) PAR + UVA (UVA) and (iii) PAR + UVA + UVB (UVR). Tropical Chlorella showed better adaptive ability to UVA stress compared to strains from temperate and polar regions. UVB stress caused significant photosynthetic dysfunction in all samples, with polar strains showing a lower inhibition (about 40%) compared to the tropical strain (about 98%). Photosynthetic responses in Chlorella towards UVR are possibly origin dependent. DNA mutation induced by both UVA and UVR treatments was revealed by Random Amplified Polymorphic DNA (RAPD) analysis. Out of sixty RAPD primers tested, two primers: S33 (polymorphism degree 44.83%) and S90 (polymorphism degree 38.71%) were chosen as potential primers to conduct genomic study of UV stress in microalgae.

Genetic Diversity of Vernonia as Revealed By Random Amplified Polymorphic DNA (RAPD) Markers

Vernonia Schreb. is a genusin the family Asteraceae. It has over 1000 species which may be trees, shrubs,woody climbers or herbs. Some of the species are economically important assources of food and herbal medicine as well as for industrial purposes. Thisstudy was carried out to assess the genetic diversity of Vernoniaspecies in Nigeria. Molecular characterization was carried out on a total of 50Vernonia samples consisting of 34 samples (13 species) from fieldcollections and 16 samples (10 species) from Ife Herbarium collections making atotal of 23 species using 4 RAPD primers. Analysis of the molecular data wasdone using NTsys version 2.02j computer programme. Results from molecularstudies showed amplification through polymerase chain reaction (PCR) with 4RAPD markers namely OPA-02, OPD-14, OPC-10 and OPE-01. Analysis of themolecular data grouped the samples into 4 Groups with only one speciesungrouped. Group I had 17 species, II had 3 species, III had 6 species while IVhad 2 species. V. amygdalina wasrepresented in all 4 Groups, V. glabra and V. guineense werepresent in Groups 1 and 3, V. tenoreana and V. purpurea werein Groups 1 and 2 respectively, while V.colorata remained ungrouped. All 50 Vernonia samples used in themolecular study clustered at 64% and analyzed at a similarity coefficient of72%. This analysis revealed that RADP markers are useful tools in assessing thelevel of genetic diversity in Vernonia.

DETERMINATION OF GENETIC VARIATION OF EGYPTIAN COWPEA (VIGNA UNGUICULATA (L.) WALP.) USING RAPD-PCR PATTERNS

Cowpea (Vigna unguiculata (L.) Walp.) is the most important grain legume crop grown in tropical and subtropical regions. Seeds of cowpea have a high nutritional value containing high amount of protein (23–29%). Seeds of 19 landraces of cowpea (Vigna unguiculata) were obtained from gene bank of Cairo University (11 landraces) and from Agricultural Research Station Kafr El-Sheikh (Sakha) (8 varieties). The seeds planted till maturity. Random amplified polymorphic DNA (RAPD) was assayed to determine the genetic variation of 19 varieties of cowpea cultivated in different regions of Egypt. A total of five random primers were used in the study. Amplification of genomic DNA of Egyptian cowpea landraces with these RAPD primers yielded 54 fragments that could be scored, of which 45 were polymorphic. Number of amplified fragments with random primers ranged from 4 (OP 856) to 14 (OP 880). Percentage polymorphism ranged from 75% (OP 856) to a maximum of 100% (OP 889, OP 868, Op 808 and OP 880), with an average of 95%. A dendrogram produced by the Unweighted Pair Group Method with Arithmetic Mean (UPGMA) based on Jaccard‟s similarity coefficient revealed that the cowpea varieties divided into two groups which showed variability that may be exploited for crop improvement. The overall range of genetic similarities ranged from 0.48 to 0.93 in 19 landraces of cowpea which indicates there was high variability among the varieties. Based on genetic distance in RAPD analysis the landraces no. 9 and 12 appeared as most divergent and could be used in breeding programme of cowpea. It could be concluded that RAPD markers are important tools for genetic analysis for cowpea(Vigna unguiculata (L.) Walp.).

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.

Characterization of Acidovorax avenae subsp. avenae causing bacterial leaf streak of maize in Punjab state of India

Twenty five isolates of Acidovorax avenae subsp. avenae causing bacterial leaf streak collected from infected maize crop grown in different geographical regions of Punjab, northern India were characterized for their molecular diversity using twenty five random amplified polymorphic DNA (RAPD) markers. All the primers showed amplification with a total of 1769 amplified fragments. Among RAPD primers, S112 was found to be highly polymorphic with PIC value of 0.95 while OPT-4 was least polymorphic. The size of amplified DNA fragments ranged from 59 bp to 3.9 kbp. Dendrogram based on molecular data generated by 25 RAPD collectively divided all 25 isolates into four groups. From 25 isolates, 20 were used for their pathological characterization and based on the multivar cluster analysis of pathogenicity data, these were divided into three groups, each producing a distinct range of disease score on particular maize inbred/hybrid used in this study. Isolate Aaa2 was found to be the most virulent with an average disease score of 8.6, whereas Aaa17 was found least virulent with average disease score of 5.1. The most virulent isolates of this pathogen were present in central Punjab. This was the first attempt to study the genetic diversity and pathogenic variation in Punjab populations of A. avenae subsp. avenae.

Estimating the Genetic Relationship in Various Warm Season Turfgrass Genotypes Using RAPD and ISSR Markers

The genetic diversity of 13 warm season turfgrass genotypes were studied in which 7 genotypes is of Cynodon dactylon and one each of C. dactylon × C. transvaalensis, Zoysia matrella, Zoysia tenuifolia, Stenotaphrum secundatum, Axonopus compressus and Eremochola ophiuroides were assessed using PCR based Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeat (ISSR) markers. Out of 150 primers of RAPD and 18 primers of ISSR screened, six each of RAPD and ISSR primers, were used to generate polymorphism for all genotypes. The dendrogram obtained from UPGMA cluster analysis of Jaccard’s similarity values ranged from 0.029 to 0.395 for RAPD and from 0.085 to 0.750 for ISSR primers. Based on above markers the genotypes were grouped into six distinct clusters.

DNA polymorphism analysis of Indian germplasms of Trapa natans using RAPD molecular Marker

Abstract Water Chestnut (Trapa natans) is a starch producing annual hydrophytic plant with potential as a secure food source. Breeding programmes mostly rely on the DNA polymorphism present in the primary gene pool. Knowledge of the genetic structure for the population is limited, and there are very few reports available on the DNA polymorphism of T. natans. In this study, we have screened RAPD based marker to estimate the DNA polymorphism among 14 germplasms of water chestnut. 180 random amplified polymorphic DNA (RAPD) marker were tested to investigate DNA polymorphism among 14 germplasms of T.natans. Out of 180 RAPD primers, 16 primers gave reproducible bands and are selected for further investigation. A total number of 220 bands were detected, among which 151 bands were polymorphic. The percentage polymorphism varied from 37.5 to 93.75. The average polymorphism across all the fourteen germplasms was found to be 68.63%, while average number of polymorphic band was 9.4. Jaccard's similarity coefficient ranged from 0.45 to 0.93. The analysis of DNA polymorphism is useful for crop improvement and may be helpful for future plant breeders.

Evaluation of Genetic Diversity among Persian Fig Cultivars by Morphological Traits and RAPD Markers

Ficus carica L. is one of the most ancient fruit trees cultivated in Persia (Iran). The conservation and characterization of fig genetic resources is essential for sustainable fig production and food security. Given these considerations, this study characterizes the genetic variability of 21 edible F. carica cultivars in the Fars Province using random amplified polymorphic DNA (RAPD) markers. The collected cultivars were also characterized for their morphological features. A total of 16 RAPD primers produced 229 reproducible bands, of which, 170 loci (74.43%) were polymorphic with an average polymorphic information content (PIC) value of 0.899. Genetic analysis using an unweighted pair-group method with arithmetic averaging (UPGMA) revealed genetic structure and relationships among the local germplasms. The dendrogram resulting from UPGMA hierarchical cluster analysis separated the fig cultivars into five groups. These results demonstrate that analysis of molecular variance allows for the partitioning of genetic variation between fig groups and illustrates greater variation within fig groups and subgroups. RAPD-based classification often corresponded with the morphological similarities and differences of the collected fig cultivars. This study suggests that RAPD markers are suitable for analysis of diversity and cultivars’ fingerprinting. Accordingly, understanding of the genetic diversity and population structure of F. carica in Iran may provide insight into the conservation and management of this species.

Identification of Bactrian camel cell lines using genetic markers

Iranian Bactrian camel population is less than 100 animals. Iranian biological resource center produced more than 50 Bactrian camel fibroblast cell lines as a somatic cell bank for conservation animal genetic resources. We compared two type markers performance, including 14 random amplified polymorphic DNA (RAPDs) (dominant) and eight microsatellite (co-dominant) for cell line identification, individual identification and investigation genetic structure of these samples. Based on clarity, polymorphism, and repeatability, four RAPD primers were selected for future analysis. Four RAPD primers and eight microsatellite markers have generated a total of 21 fragments and 45 alleles, respectively. RAPD primers revealed fragment size between 150 to 2000bp and gene diversity since 0.27 (IBRD) to 0.46 (GC10), with an average of 0.37. Microsatellite markers generated number of alleles per locus ranged from 3 to 11, with an average of 5.62 alleles. The observed heterozygosity ranged from 0.359 (IBRC02) to 0.978 (YWLL08), and expected heterozygosity ranged from 0.449 (IBRC02) to 0.879 (YWLL08). Bottleneck analysis and curve showed that Bactrian camel population did not experience a low diversity. RAPD profiles were especially suitable for investigation population genetics. All primers generated novel and polymorphic fragments. Briefly, our results show that a multiplex PCR based on these markers can still be valuable and suitable for authentication of cell lines, investigating gene diversity and conservation genetic resources in Bactrian camel, while new technologies are continuously developed.

First report of Varroa genotype in western Asia based on genotype identification of Iranian Varroa destructor populations (Mesostigmata: Varroidae) using RAPD marker

Random amplified polymorphic DNA (RAPD) technique was used to the analysis of the haplotypes of Varroa destrcuctor in southern half of Iran. Eight Varroa populations were collected and five RAPD primers were used to determine their genotype. All samples showed the K pattern genotype. Furthermore 30 loci were amplified; from which 29 were polymorphic (96.67%) and 1 was monomorphic (3.33%). This is the first molecular genetic study of V. destructor in Iran. Detailed investigations at DNA sequence level are needed to characterize genetic structure of V. destructor populations in Iran.

ANALISA GENETIK PISANG HIBRID DIPLOID BERDASARKAN MARKA RAPD (Random Amplified Polymorphic DNA)

Banana breeding by crosses is one way to get new cultivars with a better quality. This study aims to verify molecular characteristic of diploid hibrid bananas i.e MDRK, SNMT and MDMT using Random Amplified Polymorphic DNA (RAPD) markers. Ten RAPD primers have been selected to detect the differences in molecular pattern from all three diploid hybrid bananas. The results showed that the hybrid combination of MDRK, SNMT and MDMT produced bands that shared some similarities with each parents by 43.83%, 51.25% and 47.21% respectively as the number of bands from type I, III and IV markers. Type III marker is the important marker for identification of male parents. Type VII marker (non-parent bands) was detected from hybrid MDRK, SNMT and MDMT by 18 (20.22%), 6 (7.5%) and 11 (12.64%) respectively. Marker of type VII can be used to identified hybrid because they are present only in offspring. The highest polymorphic bands was obtained by SNMT (83.9%) and the lowest obtained was performed by MDMT (71.25%). From this result, RAPD markers can be used as a tool to detect the differences of molecular pattern of diploid hybrid bananas.

Genetic Diversity in Cultured and Wild Population of Clarias gariepinus (Burchell, 1822) in Nigeria Using Random Amplified Polymorphic DNA (RAPD) and Microsatellite DNA

The population structure and genetic diversities of Clarias gariepinus from the cultured population at Chi Farm (Ajanla) and wild population at Asejire Reservoir (Asejire) were analysed using Random Amplified Polymorphic DNA (RAPD) and Microsatellite DNA markers. Using a CTAB protocol, genomic DNA was extracted from the caudal fins of 20 samples of live specimen collected from each population. Seven RAPD primers and seven pairs of microsatellite DNA primers were used to amplify different loci on the extracted genomic DNA by Polymerase Chain Reaction and the resultant DNA fragments were analysed on agarose gel. The RAPD primers amplified a total of 474 loci with 697 bands in all samples for the seven primers studied. The cultured population from Chi farm showed a total of 366 bands, while the wild population from Asejire Reservoir displayed 331 bands. The cultured population showed a negative inbreeding coefficient (F) of -0.173 ± 0.209, which statistically suggests excess heterozygosity, while a positive but low inbreeding coefficient of 0.042 ± 0.243 was estimated for the wild population. The Analysis of Molecular Variance (AMOVA) for both genetic markers indicated significant difference (p=0.01) between the two populations. The result of the study suggests a loss or on-going loss of genetic variability, which needs conservation intervention in the two populations studied.

Taxonomic variation among <i>Schinus molle</i> L. plants associated with a slight change in elevation

This study examined the degree of variations in DNA fingerprints associated with slight altitudinal change of Schinus molle grown in Abha region, Saudi Arabia. Seven populations from Schinus molle plants located at 2193.0, 2246.0, 2197.7, 2441.0, 2372.0, 2250.6 and 2175.0 meters had been investigated. The degree of genetic variability was evaluated using random amplified polymorphic DNA (RAPD), mixed RAPD and intersimple sequence repeat markers (ISSR). The genetic similarity coefficients from RAPD analysis revealed the maximum similarity value (89.9%) was between population at 2250.6 m and population at 2175.0 m. The genetic similarity coefficients from mixed RAPD primers displayed the highest similarity value (87.6%) between population at 2246.0 m and population at 2197.7 m. Similarity coefficients from ISSR analysis revealed the highest similarity value (86.2%) among populations at 2193.0 m, 2246.0 m, 2441.0 m and at 2250.6 m. Super tree analysis (RAPD + mixed RAPD + ISSR) showed the highest similarity value (85.5%) between population at 2441.0 m and population at 2250.6 m. In conclusion, marker systems including RAPD, mixed RAPD and ISSR, alone or combined can be effectively used in determining the genetic relationship among Schinus molle plants even at very close populations.Bangladesh J. Plant Taxon. 24(2): 205–214.

Genetic diversity within and among two-spotted spider mite resistant and susceptible common bean genotypes

Two-spotted spider mite (Tetranychus urticae C. L. Koch, 1836), is one of the most destructive herbivores of common bean. Very little is known about the diversity among resistant sources in this crop. The present study was conducted to characterize 22 resistant and susceptible common bean genotypes by 8 Simple Sequence Repeats (SSRs) and 8 Random Amplified Polymorphic DNA (RAPD) markers. These SSR and RAPD primers produced 100 % and 81.8 % polymorphic bands. Based on RAPD fingerprints and SSR profiles, pairwise genetic similarity ranged from 0.0 to 0.857 and from 0.125 to 1, respectively. The resistant and susceptible common bean accessions were grouped together in the dendrograms generated from RAPD and SSR clustering analyses. The results indicate that RAPD and SSR analysis could be successfully used for the estimation of genetic diversity among genotypes. SSR markers could group genotypes according to their resistibility and susceptibility to the spotted spider mite but RAPD could not. Therefore, the SSR markers can facilitate the development of resistant common bean cultivars through breeding programs against T. urticae.

Phylogenetic Relationship and Genetic Variation among Thaumatococcus daniellii and Megaphrynium macrostachyum Ecotypes in Southwest Nigeria

Background and Objective: Thaumatococcus daniellii (T. daniellii) and Megaphrynium macrostachyum (M. macrostachyum) are perennial, understorey herbs with similar morphological and vegetative properties. Genetic relatedness and variation among and within ecotypes of the plant species in Nigeria were studied using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers. Materials and Methods: Plant samples were collected at random from 24 sites across 5 states in Southwest Nigeria. The DNA was extracted from the leaves by cetyl trimethyl ammonium bromide (CTAB) method and amplified with respective PCR-based primers. Genetic variation within and between the populations was determined by analysis of molecular variance (AMOVA), cluster analysis was performed using unweighted pair group method using arithmetic averages (UPGMA). Results: The RAPD and ISSR markers generated clear reproducible bands, polymorphic bands (PPD) produced by RAPD and ISSR markers were 86.7 and 91.3, respectively. Genetic variation among the two plant populations was 93.68 and 77.87, respectively with RAPD and ISSR markers, variation within each plant population was 6.32 and 22.13, respectively for the RAPD and ISSR primers. The cluster analysis indicated that genetic differentiation occurred more among the population than within population of T. daniellii and M. macrostachyum. Conclusion: This study showed that T. daniellii and M. macrostachyum species are genetically distinct from each other. However, the genetic variation within each of the two species is low, thus, management and conservative approaches are required to preserve and develop the plants for future commercial exploitation. © 2018 Shalom Nwodo Chinedu et al.

SEX IDENTIFICATION OF DATE PALM BY USING DNA MOLECULAR MARKERS

This study was conducted to investigate the early detection of date palm gender, the current study was conducted using some DNA markers technology using Bulk Segregant Analysis (BSA) and three molecular markers including Randomly Amplified Polymorphic DNA (RAPDs), and Cleaved Amplified Polymorphic Sequence (CAPS) and Nested PCR. Results showed that RAPD primers used cleared different banding pattern. Fifty primers showed monomorphic bands while ten showed polymorphic bands for the two balks Males (MB) and females (FM). These primers showed 626 total bands in which 41 of them was polymorphic (6.54% polymorphism). OPD.10 primer gave the highest number of bands reached 118 bands and was the highest efficiency (18.85%). While the primer OPE04 was least efficient (3.51%), giving only 22 bands. A band with molecular weight of 143 bp appeared in six females of the total seven female varieties (85.71%) and did not appear in all males suggesting a positive marker which could be consider a candidate region for gender in date palm. Results of CAPS markers using three restriction enzymes: Bcl I, Hpa II and RsaI to digest the PCR products of OPD.10 primer. Results indicated that not all enzymes were useful for cutting DNA of samples. Bcl I enzyme cut the band of 143 bp which appeared in six female samples, while Hpa II enzyme cut the band of 180 bp in five males and six females. The former enzyme had also cut and the band of 433 bp in all female varieties. No response was noted when DNA was restricted with enzyme RsaI. The above results consider to be promising for gender detection in date palm.

Genetic diversity in kiwifruit germplasm evaluated using RAPD and SRAP markers

In this study, random amplified polymorphic DNA (RAPD) and sequence-related amplified polymorphism (SRAP) analyses were used for evaluation of genetic diversity of 61 kiwifruit (Actinidia spp.) germplasms including domestic and overseas collection cultivars. Forty RAPD primers were detected in a total of 230 polymorphic bands with an average of 5.75. Thirty-two SRAP primer combinations were detected in a total of 204 polymorphic bands with an average 6.38. By unweighted pair-group method arithmetic average cluster analysis using 434 polymorphic bands, kiwifruit germplasms were classified in three groups with similarity value of 0.680. Cluster I consisted of 46 kiwifruit germplasms belonging to A. deliciosa, A. chinensis, A. deliciosa × A. arguta, A. chinensis × A. arguta, and A. chinensis × A. deliciosa. Cluster II consisted of seven germplasms belonging to A. arguta and ‘Skinny Green’, a cultivar derived from a cross between A. arguta and A. deliciosa. Cluster III consisted of seven germplasms belonging to A. rufa, A. hemsleyana, A. macrosperma, A. polygama, and A. eriantha. Genetic similarity values among tested kiwifruit germplasms ranged from 0.479-0.991, and average similarity value was 0.717. Similarity value was highest (0.991) between NHK0038 (A. deliciosa) and NHK0040 (A. deliciosa), and lowest (0.479) between ‘Hayward’ (A. deliciosa) and K5-1-22 (A. arguta).

Evaluation of genetic fidelity among micropropagated plants of Salvadora persica L. using DNA-based markers

Salvadora persica L. (family Salvadoraceae) is a highly threatened medicinally and economically important arid zone plant species. With ever increasing demand, there is a pressing need for conservation and mass multiplication of this plant through biotechnological approaches. A large number of complete plantlets were obtained by direct regeneration from the nodal explants on Murashige and Skoog (MS) medium supplemented with 2.0mg/l 6-benzylaminopurine (BAP)+1.0mg/l indole-3-acetic acid (IAA) containing 30g/l sucrose and 8g/l agar at 16h photoperiod. A high frequency of in vitro rooting was obtained on half strength MS medium supplemented with 3.0mg/l indole-3-butyric acid (IBA). The micropropagated plantlets were hardened using sterilized soil and sand mixture (3:1) and successfully transferred under field conditions. The genetic stability of micropropagated clones was evaluated using random amplified polymorphic DNA (RAPD) analysis. During the study a total of 43 RAPD primers were screened, out of which 8 primers produced a total of 37 clear, distinct and reproducible amplicons. The amplified products of all the regenerated plants were monomorphic and were similar to the mother plant, thus confirming the genetic homogeneity of tissue culture raised plants. To the best of our knowledge, it is the first report on the assessment of the genetic fidelity in micropropagated plants of S. persica, which could be successfully applied as one of the safest modes for the production of true-to-type plants for the germplasm conservation.