RAPD Molecular Markers Research Articles

Development of SCAR molecular markers for early and late differentiation of Eucalyptus globulus ssp globulus from E. globulus ssp maidenii

The genus Eucalyptus includes over 700 species, some of which are the most widely planted hardwoods worldwide. Each species and subspecies of Eucalyptus present different characteristics regarding its wood quality and yield. This fact makes it very important to work with known species/subspecies so as to optimize handling and conservation of forest resources. Some of them are morphologically very similar, making it difficult to differentiate by simple observation. This is the case with Euclyptus globulus ssp globulus and E. globulus ssp maidenii, which can only be distinguished in the adult tree. These issues can be avoided using well-characterized seeds. This can be quite expensive, especially for the small growers. An alternative approach is to develop simple methods for the differentiation of the two subspecies. In this work, we develop a quick method, based on SCAR molecular markers derived from RAPD molecular markers, for the differentiation of the subspecies of E. globulus, in particular E. globulus ssp globulus and E. globulus ssp maidenii.

Genetic diversity of the species Leporinus elongatus (Teleostei: Characiformes) in the Canoas Complex - Paranapanema River

Dams constructed along waterways interrupt the dispersion and migration of aquatic organisms, affecting mainly the abundance of migratory fish species. Translocation mechanisms have been constructed at dams aiming to minimize their impact on fish species migration behavior. There is little information available about the effect of the construction of dams on the genetic structure of the Neotropical migratory fish fauna. Therefore, RAPD molecular markers and microsatellites were utilized to evaluate the diversity and genetic structure of the migratory species Leporinus elongatus (piapara) in the Canoas Complex - Paranapanema River - Brazil. Ten groups were sampled in the fish ladders of the hydroelectric dam Canoas I and Canoas II during the reproductive period in three consecutive years. Both markers showed a high level of genetic diversity within these groups. The microsatellite markers demonstrated a loss of heterozygosity and a considerable level of inbreeding in the species. The genetic differentiation found among the groups with both markers utilized is within a range from low to moderate. The data obtained with the parameter of genetic diversity among the groups led to the conclusion that the groups of L. elongatus of the Canoas Complex are structured as a single population composed of sub-populations with low genetic diversity among them. The data on genetic diversity and population structure of L. elongatus are of great importance for the development of the species management and conservation programs in the Canoas Complex, which can also be utilized in aquaculture programs.

Evaluation of genetic diversity among some genotypes of Kentucky bluegrass by RAPD molecular markers

Genetic improvement of plants need a high genetic diversity and molecular markers offer a powerful tool for its assessment. Poa pratensis has extensive divergence in natural old turfs and pastures in temperate zones of Iran, being useful for breeding programs. In this research, RAPD markers were used to study the genetic diversity of 19 turfgrass genotypes (including 16 genotypes of Poa pratensis along with one genotype from each of Lolium perenne, Cynodon dactylon and Festuca arundinacea species). After screening 100 random 10-mer RAPD primers, 17 primers were selected based on the clarity of their amplified fragments in PCR and the high reproducibility for scoring. From the 284 produced bands, 281 were polymorphic. Cluster analysis of RAPD data was performed using the UPGMA method based on Jaccard’s similarity coefficients. Dendrogram at a similarity of 0.24 gave 5 main clusters. The correlation coefficient between the data matrix and the cophenetic matrix of cluster data was high (r = 0.99), indicating that the clustering dendrogram being highly fitted to similarity matrix. A relatively high genetic similarity was observed between some commercial cultivars and Iranian local genotypes. In addition, this research showed the high genetic diversity among studied genotypes as well as the high efficiency of RAPD markers for evaluation of genetic diversity among turfgrasses.

MORPHOLOGICAL AND MOLECULAR GENETICS CHARACTERIZATION OF HOLOPARASITIC PLANT, Cistanche phelypea L. IN SIWA OASIS, EGYPT

Cistanche phelypea (Fam. Orbanchaceae) is a parasite plant on the root of the host plant. The main natural hosts are the woody species of family chenopodiaceae such as Arthrocnemum macrostachyum and family Zygophyllacea such as Zygophyllum album. It is an obligate parasite which totally depends on its host for water, minerals and organic nutrients. Many species belonging to the same family have the same phenomenon such as orbanche crenata Forsk which attacks legume plants, specially, Vicia faba. Cistanche phelypea L., grows in Egypt in low density but scattered on a large area of the country. It can be recognized in the Nile region including Faiyum, the Oases of the western desert including Siwa, the Red Sea coastal strip, and the entire Sinai Peninsula including the coastal of the Mediterranean. It grows in sandy and alluvial soil, edges of cultivated fields as a parasite on plant roots. In this study the species was surveyed and photographed in Siwa Oasis and in different obliterated Oasis around Siwa Depression. For the first time in Egypt, morphological, chemical study, RAPD molecular markers and biochemical genetic analysis of Peroxidase isozymes were carried out in order to identify the genetical and morphological description of the species including the identification of genetic variation in the studied populations.

Characterization of the genetic variability among Caesalpinia pulcherrima (L.) Sw. (Fabaceae) plants using RAPD molecular markers

The species Caesalpinia pulcherrima, which belongs to the Fabaceae family, is frequently used as a hedge or street tree in urban landscapes, parks and gardens. It flowers profusely, and the flowers, depending on the variety, may be pink, orange or yellow. As characteristics are essential for determining the commercial value of ornamental species, the main objective of this work was to identify polymorphisms in C. pulcherrima plants that produce flowers of different colors using RAPD molecular markers. For this study, 30 adult plants were randomly chosen on streets located in Jaboticabal city, Sao Paulo State, Brazil, and leaf samples were collected for DNA extraction. Among these trees, 20 have orange flowers, eight have yellow flowers and only two produce pink flowers. From the 140 tested RAPD primers, 94 primers amplified defined fragments that resulted in 246 bands, which were 100% monomorphic. Thus, a polymorphism was not detected by any of the primers tested. It was concluded that the RAPD technique is not an efficient method for detecting polymorphisms and that more specific molecular markers should be tested. Additionally, the morphological characteristic flower color may be controlled by several genes or by the association of them.

Discrimination Capacity of RAPD, ISSR and SSR Markers and of their Effectiveness in Establishing Genetic Relationship and Diversity among Egyptian and Saudi Wheat Cultivars

Problem statement: Yield crop cultivars and landraces are valuable sources of genetic variations that the knowledge and implication of these variations are critical in the plant breeding programs. our major objective of this study is investigating the discriminating capacity of RAPD, ISSR and SSR markers and of their effectiveness in establishing genetic relationship and diversity among Egyptian and Saudi wheat cultivars. Approach: Eleven wheat cultivars and landraces collected from Egypt and Saudi Arabia, five Egyptian wheat (Sakha 93, Sods 1, Sods 4, Gmiza 9 and Sohag 3) and six Saudi wheat landrace cultivars (Hmees, Al-Kaseem, Hegazi, Abo-Sakr, Dubai 1 and Nagran) were characterized using RAPD, ISSR and SSR molecular markers as efficient tools. Ten and nine oligonucleotide primers of RAPD and ISSR respectively and four primer pairs of SSR were used in wheat samples analysis. Only clear and repeatable band profile of 6 RAPD, 8 ISSR and 2 SSR primers were obtained. In RAPD analyses, 74 out of 141 bands (52%) were polymorphic. Results: The number of alleles ranged from 8-21 per primer, with an average of 14.1 per primer. In ISSR analyses, a total of 78 alleles were detected, along with 36 alleles (46%) were polymorphic. The number of alleles per primer ranged from 5-10 with an average of 8.6 alleles per ISSR primer. SSR reactions recorded 6 alleles, of which 5 alleles (83%) were polymorphic. Cluster analysis was conducted using Unweighted Pair Group Method that depends on Arithmetic Average (UPGMA). The dendrogram cluster diagram classified the evaluated genotypes in three major clusters corresponding to the cultivation regions. The first group contains Sakha 93, Sods 1 and Sods 4 with more than 80% Genetic Similarity (GS). The GS between Sakha 93 and Sods 1, Sakha 93 and Sods 4 or Sods 1 and Sods 4 were 83.6%, 83.9 and 85.4 respectively. The second group contains Gmiza 9 and Sohag 3 with GS 83.1%. The third group contains most of the Saudi landrace cultivars, Hmees, Al-Kaseem, Dubai 1, Abo-Sakr and Nagran, which are genetically closed to each other with GS of 81%. The last Saudi wheat landrace cultivar, Hegazi, was falling outside the three major clusters, revealing around 78% similarity with the rest of the five Saudi landrace cultivars. Conclusion/Recommendations: These analyses fit together with geographical distribution of the 11 wheat cultivars and landraces. Moreover, some morphological characterizations as fresh and dry or flowering time between the selected cultivars were analyzed under different salt concentration. We recognized differences in the fresh and dry weight between the selected cultivars. Wheat cultivars Sods 4 and Sohag 3 were the most sensitive cultivars to the salt treatment, while Sods 1 and Sakha 93 cultivars were less sensitive to the salt treatments. Additionally, Sods 4 and Sakha 93 cultivars were the earliest among the five wheat cultivars (flowering time 68.0±5.04 and 71±6.97 days respectively), while Sohag 3 and Gmiza 9 have flowered later than the other five cultivars (111±12 and 105±11.1) respectively. In conclusion, the long term objective of this study was to use these fingerprints to identify molecular markers that co-segregate and could be used in isolating gene(s) which controlling some important traits.

A genetic linkage map of crested wheatgrass based on AFLP and RAPD markers

Using a population of 105 interspecific F(2) hybrids derived from a cross between Agropyron mongolicum Keng and Agropyron cristatum (L.) Gaertn. 'Fairway' as a mapping population, a genetic linkage map of crested wheatgrass was constructed based on AFLP and RAPD molecular markers. A total of 175 markers, including 152 AFLP and 23 RAPD markers, were ordered in seven linkage groups. The map distance was 416 cM, with a mean distance of 2.47 cM between markers. The number of markers ranged from 13 to 46 in each linkage group and the length of groups ranged from 18 to 104 cM. The research found that 30 out of 175 molecular markers showed segregation distortion, accounting for 17% of all markers. This is the first genetic linkage map of crested wheatgrass. This map will facilitate gene localization, cloning, and molecular marker-assisted selection in the future.

A comparison of RAPD and ISSR markers reveals genetic diversity among sweet potato landraces (Ipomoea batatas (L.) Lam.)

Small farmers have played an important role in maintaining genetic resources of traditional varieties. Collecting and characterizing germplasm, especially landraces and heirloom varieties, are strategic activities that would create a knowledge base describing the traditional varieties of a given region and reduce the risk of genetic erosion. The objectives of this work were as follows: collect sweet potato roots from rural properties and local markets in Rio de Janeiro State, Brazil; characterize the accessions using RAPD and ISSR molecular markers, comparing the efficiency in discrimination of these two types of molecular markers; and estimate the genetic variability of the population. Fifty-nine samples from rural properties and 19 from local markets were studied. A matrix of binary data was developed to analyze the molecular data using the Jaccard index to estimate genetic differences among accessions, and clusters were made using the UPGMA method. The molecular techniques detected genetic variability among the accessions, with cophenetic correlation coefficients (CCC) of 0.80 for RAPDs and 0.89 for ISSRs. The RAPD and ISSR marker results corresponded well, with a correlation of 0.55. All of the accessions were considered distinct, which demonstrates that traditional farmers maintain sweet potato genotypes that have good genetic diversity.

Diversity Analysis of Tomato Cultivars Based on Coefficient of Parentage and RAPD Molecular Markers

Genetic diversity analysis provides information on the genetic base of the gene pool of released genotypes. The objectives of this study were to assess the diversity of tomato cultivars and breeding lines released from North Carolina State University based on their coefficient of parentage (COP) and RAPD markers, and to estimate the contribution of ancestors to these tomato cultivars and breeding lines. The COP analysis revealed that a single ancestor Walter constituted 25.17% of the North Carolina tomato gene pool followed by Blazer (11.57%) and Fla. 2153 (6.29%). Walter was used in developing 30 lines, whereas Blazer and Fla. 2153 were used in developing 29 lines each. The highest COP value (0.828) was between Summit and Monte Verde, indicating that they were the most similar lines. Similarity coefficients based on the RAPD molecular markers ranged from 0.461 to 0.935, with an average of 0.77. The similarity coefficient of NC3Grape with NCEBR8 was the lowest (0.461), whereas it was the highest (0.935) between NC1CS and NC946. On the basis of molecular data, first and second principal components accounted for 78% and 3.3% of the total variation, respectively. A significant positive correlation (r = 0.19, p = 0.04) existed between two similarity matrices based on the RAPD and the pedigree. This information may be useful for selecting the parents in breeding programs, particularly to widen the genetic base for designing future breeding strategies.

Genetic Distance Estimated by RAPD Markers and Performance of Topcross Hybrids in Popcorn

The purpose of this study was to investigate the diversity of 14 popcorn populations and a broad genetic base tester using molecular RAPD markers and to estimate the correlation between the genetic distances and the performance of top-cross hybrids. For the evaluation of populations and hybrids resulting from topcrosses, the reduced model of Gardner was used. A genetic distance matrix was generated based on RAPD markers by Jaccard coefficient, and a dendrogram was constructed. In general, topcrosses performed better than the populations per se and evidenced heterosis occurrence in topcrosses. The trait grain weight is influenced by additive as much as by dominance effects. Genetic associations separated the populations in three groups, and RAPD showed to be a useful tool to determine the extension of genetic diversity in popcorn populations and to place genotypes in distinct heterotic groups. Correlations between genetic divergences, detected by RAPD, and the means observed in the topcross crosses were positive and non-significant for expansion volume, plant height, and female flowering, and were negative for grain weight.

A Comprehensive Analysis of Genetic Diversity and Relationships of 13 Sweet Sorghum Varieties as Energy Sources

To investigate the genetic diversity and relationships among the sweet sorghum varieties as energy sources currently bred in China, 13 sweet sorghum varieties were selected for comprehensive analysis through observations of 31 biological traits and examinations of RAPD and SSR molecular markers. The numerical analysis showed that the differences in biological traits existed among 13 varieties, and the genetic distance (DIST) ranged from 0.787 to 2.221, and the two varieties from Inner Mongolia and Xinjiang were distinctly separated from all other varieties. A total of 22 polymorphism primers were obtained from the screening using RAPD marker analysis. The polymorphism rate was 58.33%, and the genetic similarity (GS) coefficients among the studied cultivars ranged from 0.694 to 0.896. Cluster analysis results indicated that the three varieties from Inner Mongolia, Xinjiang and Heilongjiang exhibited significant genetic differences from the other varieties. SSR marker analysis using 31 selected pairs of polymorphic primers showed that the polymorphism rate of amplified fragments was 78.64%, and GS coefficients among the tested cultivars were 0.534 to 0.971. Cluster analysis showed that variety No. 12 from Xinjiang and variety No. 7 from Inner Mongolia clustered into one group, and variety No. 6 from Heilongjiang was in a single group. The other ten varieties were grouped into another separate cluster. The results based on combined data displayed a similar trend with results from the three individual data analyses, but could more comprehensively and objectively reflect the fundamental genetic differences among these varieties. In summary, certain genetic differences exist among the varieties tested from different regions or different breeding institutions. However, varieties from the same region, especially those from the same breeding institution, exhibited small genetic variations and high genetic similarities. At present, more attention should be paid to discovery and innovation in the breeding of sweet sorghum varieties.

Genetic characterization of fig tree mutants with molecular markers

The fig (Ficus carica L.) is a fruit tree of great world importance and, therefore, the genetic improvement becomes an important field of research for better crops, being necessary to gather information on this species, mainly regarding its genetic variability so that appropriate propagation projects and management are made. The improvement programs of fig trees using conventional procedures in order to obtain new cultivars are rare in many countries, such as Brazil, especially due to the little genetic variability and to the difficulties in obtaining plants from gamete fusion once the wasp Blastophaga psenes, responsible for the natural pollinating, is not found in Brazil. In this way, the mutagenic genetic improvement becomes a solution of it. For this reason, in an experiment conducted earlier, fig plants formed by cuttings treated with gamma ray were selected based on their agronomic characteristics of interest. We determined the genetic variability in these fig tree selections, using RAPD and AFLP molecular markers, comparing them to each other and to the Roxo-de-Valinhos, used as the standard. For the reactions of DNA amplification, 140 RAPD primers and 12 primer combinations for AFLP analysis were used. The selections did not differ genetically between themselves and between them and the Roxo-de-Valinhos cultivar. Techniques that can detect polymorphism between treatments, such as DNA sequencing, must be tested. The phenotypic variation of plants may be due to epigenetic variation, necessitating the use of techniques with methylation-sensitive restriction enzymes.

Genetic similarity among strawberry cultivars assessed by RAPD and ISSR markers

Most strawberry (Fragaria × ananassa Duchesne) cultivars used in Brazil are developed in other countries, it became clear the need to start the strawberry breeding program in the country. To start a breeding program is necessary the genetic characterization of the germplasm available. Molecular markers are important tools that can be used for this purpose. The objectives of the present study were to assess the genetic similarity among 11 strawberry cultivars using RAPD and ISSR molecular markers and to indicate the possible promising crosses. The DNA of the eleven strawberry cultivars was extracted and amplified by PCR with RAPD and ISSR primers. The DNA fragments were separated in agarose gel for the RAPD markers and in polyacrylamide gel for the ISSR markers. The genetic similarity matrix was estimated by the Jaccard coefficient. Based on this matrix, the cultivars were grouped using the UPGMA method. The dendogram generated by the RAPD markers distributed the cultivars in three groups while the ISSR markers generated two groups. There was no direct relationship between the marker groups when the two types of markers were compared. The grouping proposed by the ISSR markers was more coherent with the origin and the genealogy of the cultivars than that proposed by the RAPD markers, and it can be considered the most efficient method for the study of genetic divergence in strawberry. The most promising crosses, based on the genetic divergence estimated from the RAPD and ISSR molecular data were between the Tudla and Ventana and the Oso Grande and Ventana cultivars, respectively.

Genetic diversity assessment of Gymnema sylvestre (Retz.) R. Br. ex Sm. populations from Western Ghats of Maharashtra, India

Gymnema sylvestre (Retz.) R. Br. ex Sm., a rich source with gymnemic acid, has been used to treat diabetes since past 2000 years. In the current investigation, nomenclature, geographical distribution as well as morphological and genetic diversity of 22 G. sylvestre populations spread across of Western Ghats of Maharashtra has been reported. The genetic variability has been assessed using RAPD and ISSR molecular markers. The populations recorded high level of gene differentiation (GST = 0.41 based on ISSR) and gene flow (Nm = 0.92 based on RAPD). The high level of gene differentiation among the populations attributed to entomophilous pollination, wind seed dispersal, out crossing and anthropogenic activities.

Genetic Diversity Analysis of Maize Inbred Lines from SCDA Turda - Romania Revealed by RAPD Molecular Markers

Knowledge of genetic diversity and relationships among maize inbred lines is indispensable to a breeding program. Our objective was to investigate the level of genetic diversity among maize inbred lines. Eighty-three maize inbred lines obtained from SCDA Turda were genotyped using 20 decamer primers. These primers generated, among the studied genotypes, a number of polymorphic bands comprised between 17 bands (OPA 03) and 7 bands (OPAB 11). The highest numbers of polymorphic bands were obtained with primer OPA 03, respectively 17 bands, followed by OPA 01, OPB 08 (16 polymorphic bands) and OPX 03 and OPAL 20 (13 polymorphic bands). Genetic distances were established using Nei Li/Dice coefficent and an UPGMA dendrogram was constructed with FreeTree software. The built dendrogram shows phylogenetic relationships between the analysed biological material.

Genetic variability of brycon orbignyanus (valenciennes, 1850) (characiformes: characidae) in cultivated and natural populations of the upper paraná river, and implications for the conservation of the species

This study aimed to evaluate the genetic variability of B. orbignyanus in cultivated and natural populations of the Upper Paraná River, using molecular RAPD markers and mtDNA control region. Specimens were collected in the Paraná River and in the Piracema fish farm in Maringá, State of Paraná, Brazil. RAPD primers produced 82 loci with consistent expression. The population from the Paraná River showed 28 polymorphic loci, whereas the population from the fish farm presented only 12. Data revealed genetic differentiation between the two populations, although not very pronounced. These results were corroborated by the principal coordinate analysis and by neighbor-joining clustering. The alignment of the D-loop sequences of B. orbignyanus indicated the existence of polymorphism only in the natural population. These data could be helpful for the formulation of management strategies and conservation of the genetic diversity of the species.

COMPARATIVE ANALYSIS OF RAPD MARKER SEQUENCES IN RELATION TO GAMMA IRRADIATION IN SESAME

Sesame (Sesamum indicum L.) is one of the most ancient crops (Bedigian et al., 1986). It is grown in tropical and subtropical areas (Ashri, 1998) on 7.7 million hectares worldwide, producing four million tons of seed (FAO, 2009). The seeds contain 50-60% oil, which is highly resistant to oxidative deterioration even though oleic and linoleic acids are the predominant fatty acids of sesame oil, about 80% of its total (Arslan et al., 2007; Uzun et al., 2007). Sesame crop is well suited to different crop rotations and is mostly grown under moisture stress with low management inputs by small holders (Ashri and van Zanten, 1994). In spite of being the first oilseed crop known to man and its long history, sesame is a typically neglected crop. It is not studied by any of the international agricultural research centers (Ashri, 1998). Genetic variation of different sesame collections has been previously reported using agro-morphological characters (Bedigian et al., 1986; Furat and Uzun, 2010; Pham et al., 2011). In addition, molecular markers, including isozymes (Isshiki and Umezaki, 1997), RAPD (Bhat et al., 1999; Ercan et al., 2004; Pham et al., 2009; Pham et al., 2011), ISSR (Kim et al., 2002), AFLP (Uzun et al., 2003; Laurentin and Karlovsky, 2006; Ali et al., 2007) and SSR (Dixit et al., 2005). Some of these techniques, such as RAPD do not require prior knowledge of DNA sequence. It has been possible to increase the genetic variability in sesame by inducing mutations with ionized radiation, allowing isolating mutants with desirable characters of economic importance such as increased seed yield, earliness (Wongyai et al., 2001), modified plant architecture, closed capsules, disease resistance (Cagirgan, 1994&2001; Ashri, 1998; Diouf et al., 2010), seed retention, larger seed size, desirable seed color and high oil content (Hoballah, 2001). RAPD analysis has been used to identify the DNA polymorphism induced by gamma rays in groundnut (Bhagwat et al., 1997), cypress (Ishii et al., 2003), soybean (Atak et al., 2004), sunflower (Erdem and Oldacay, 2004), Chrysanthemum (Lema-Ruminska et al., 2004), sugarcane (Khan et al., 2007), amla (Selvi et al., 2007), Baby's-breath (Barakat and El-Sammak, 2011) and physic nut (Dhakshanamoorthy et al., 2011) and detection of mutation in sunflower (Gunhan and Oldacy, 2004), grapes (Khawale et al., 2007) and banana (Ganapathi et al., 2008). RAPD analysis was also be used for the detection of DNA damage and mutations in young Vigna radiate calli (Roy et al., 2006). However, there are a limited number of reports on the use of RAPD markers for genetic variation studies after gamma irradiation in sesame (Mohamed et al., 1999). RAPD and ISSR molecular markers were used for tagging the dt gene regulating determinate growth habit (Uzun and Cagirgan, 2009). The only observation of DNA sequences of RAPD bands of plant species was reported by Begum et al. (2008) in Vigna radiata. The aim of the present study is to use RAPD technique for the detection of genetic polymorphism among sesame genotypes following gamma irradiation. The produced RAPD bands from control and irradiated samples were cloned and sequenced to determine possible occurrence of base pair alterations.

Chemical Polymorphism of Essential Oils from Five Populations of Thymus daenensis Celak. subsp. daenensis Endemic to Iran

Chemical composition of essential oils from five populations of Thymus daenensis Celak. subsp. daenensis, an important medicinal plant endemic to Iran, was investigated by means of GC and GC/MS. All oils were characterized by a high content of monoterpenoid compounds, especially oxygenated ones (75.4–83.0%), and a small amount of sesquiterpenes (1.7–4.0%). Thymol was identified as the major component of all essential oils. Cluster analysis based on the percentage composition of essential oils and data obtained from the RAPD molecular markers experiment led to clearly discrimination of Th. deanensis populations and they were placed into two main clusters; members of the first cluster (Arak and Malayer) contained thymol as the main oil component (77.7–78.1%), whereas the populations included in the second cluster (Aleshtar, Hamedan and Dena) were rich in carvacrol (19.1–29.4%) as well

Use of RAPD molecular markers on differentiation of brazilian and chinese Ganoderma lucidum strains

The aim of this work was to analyze the Brazilian and Chinese strains of Ganoderma lucidum with molecular RAPD markers. A similarity matrix was elaborated and the RAPD profiles of G. lucidum strains were also compared to two other Ganoderma spp: G. applanatum and G. lipsiense in order to produce genetic similarity among the species. Based on the primers used, it was possible to determine that the Brazilian strains and Chinese strain CC-22 are alike. The method and the primers selection showed to be appropriate for the genetic identification of G. lucidum strains, enabling them to be improved and used in research, as well as in the world market.

Studies on the genetic variation of the green unicellular alga Haematococcus pluvialis (Chlorophyceae) obtained from different geographical locations using ISSR and RAPD molecular marker.

Haematococcus pluvialis (Flotow) is a unicellular green alga, which is considered to be the best astaxanthin-producing organism. Molecular markers are suitable tools for the purpose of finding out genetic variations in organisms; however there have been no studies conducted on ISSR or RAPD molecular markers for this organism. The DNA of 10 different strains of H. pluvialis (four strains from Iran, two strains from Finland, one strain from Switzerland and three strains from the USA) was extracted. A genetic similarity study was carried out using 14 ISSR and 12 RAPD primers. Moreover, the molecular weights of the bands produced ranged from 0.14 to 3.4 Kb. The PCA and dendrogram clustered the H. pluvialis strains into various groups according to their geographical origin. The lowest genetic similarity was between the Iran2 and USA2 strains (0.08) and the highest genetic similarity was between Finland1 and Finland2 (0.64). The maximum numbers of bands produced by the ISSR and RAPD primers were 35 and 6 bands, respectively. The results showed that ISSR and RAPD markers are useful for genetic diversity studies of Haematococcus as they showed geographical discrimination.