Random Amplified Polymorphic DNA Molecular Markers Research Articles

Association between quantitative morphological traits and RAPD molecular markers in pomegranate (Punica granatum L.).

Pomegranate (Punica granatum L.) is very important in terms of horticulture and food around the world. The present research aimed to identify the random amplified polymorphic DNA (RAPD) markers associated with morphological traits in pomegranate genotypes. Significant differences were observed among the studied genotypes based on the recorded traits. The 18 RAPD primers produced a total of 154 polymorphic fragments among genotypes. Using multiple regression analysis between each of the morphological traits and 154 RAPD polymorphic bands, RAPD markers associated with each of the morphological traits were identified. In total, 11 markers showed significant correlations with fruit weight, 9 markers with 100-aril weight, 11 markers with anthocyanin, and 8 markers with total soluble solids. Some markers were associated with more than one morphological trait, showing that the association of a marker with more than one trait can be caused by the pleiotropic effects of quantitative trait loci related to each other in different traits. For instance, the BA6-1 marker showed positive correlations with fruit weight, fruit crown width, and leaf length. Also, OPG13-3 and BA6-10 markers showed positive correlations with total soluble solids and anthocyanin content. The informative markers identified related to morphological characteristics in pomegranate can be a suitable guide to identify the genotypes with valuable fruit traits. Also, these markers can be used in selecting suitable parents for population generation for mapping purposes.

Analysis of the Genetic Distance of Several Generations of Barley (Hordeum valulgare L) by RAPD-PCR Technique

Random-amplified-polymorphic-DNA(RAPD) was assayed to detect the genetic variation of 6 barley generations from Iraq. Four primers generated a total of 17 scoreable bands in RAPD analysis) and resolving power, the three polymorphic primers differed (Rp). The use of RAPD marker systems to detect the genetic distance among barley generation was discovered to be beneficial. The RAPD dendrograms indicate a diverse grouping of 6 barely specimens, although we did see that certain groups were identical in several cases. As a result, the RAPD molecular markers reveal two genetic groups in the few specimens examined. Keywords. Barley, Genetic variation, RAPD-PCR.

In vitro effect of phytosynthesised AgNPs to enhance plantlets and biomass production in Nyctanthes arbor-tristis L.

This work was aimed to establish an effective approach for direct regeneration of Nyctanthes arbor-tristis L. using nodal segment. Amongst the different combinations of plant growth regulators (PGR's), the maximum number of shoots/explant (32.94 ± 0.86) with 3.14 ± 0.08 cm shoot length was attained on Murashige and Skoog (MS) medium fortified with 3.5 mg /L 6-benzyladenine (BA) and 0.3 mg/L concentration of 1-phenyl-3-(1, 2, 3- thiadiazol-5-yl)- urea (TDZ). Besides, nodal segments of Nyctanthes grown on MS medium were used to analyze the effect of biogenic AgNPs (0,2,4,6,8 and 10 mg/L) on plant growth & multiplication in solid & suspension culture medium. After 35 days of in vitro shoot proliferation, the shoot number, shoot length, chlorophyll content, fresh & dry matter, guaiacol peroxidase (GPOX) and catalases enzyme (CAT) activities of plants were quantified. The administration of AgNPs (6 mg/L) along with the lower concentration of BA (1 mg/L) could enhance shoot multiplication from (1.29 ± 0.14) to (25.17 ± 0.45), chlorophyll content as well as the GPOX, and CAT enzyme activities in suspension culture medium. Optimized rooting response (12.15 ± 0.63) was noted on the half-strength MS medium supplemented with 0.5 mg/L concentration of Indole acetic acid (IAA), The maximum in vitro raised plantlets was hardened & acclimatized in a ratio of 6:2:2:1(garden soil: cocopeat: vermiculite: pearlite) potting mixture. The regenerated plantlets showed 80% survival rate under greenhouse conditions. Out of twenty PCR-based RAPD (Random Amplified Polymorphic DNA) molecular markers, 8 primers produced a total of 350 scorable bands which showed 90–99% similarity with mother plant.

Efficient micropropagation of Thunbergia coccinea Wall. and genetic homogeneity assessment through RAPD and ISSR markers

Thunbergia coccinea Wall. ex D. Don being a rare, ornamental and medicinal plant of India, is needed to propagate for conserving the germplasm and analyzing its phytochemical compounds in the future. A reliable protocol for direct in vitro propagation using nodal shoot meristem of T. coccinea as explant was standardized. The highest number of shoots per explant (22.17 ± 0.54) with maximum shoot length (2.36 ± 0.28) in cm was obtained in Murashige and Skoog (MS) medium supplemented with 9.70 µM of 6-furfurylaminopurine (Kinetin) and 0.053 µM of α-naphthaleneacetic acid (NAA) combination, among all the different plant growth regulators (PGR’s) and concentrations tested. The aforesaid PGR’s combination was optimum for axillary shoot bud induction and multiplication in T. coccinea. The best rooting was observed on the half-strength MS medium fortified with 2.68 µM NAA with the highest number of roots per shoot (3.75 ± 0.12) and maximum length (5.22 ± 0.32) in cm. All the in vitro raised plantlets were acclimatized in sterile sand and soil mixture (1:1) with a survival rate of 70% on earthen pots under greenhouse conditions. PCR-based RAPD (Random Amplified Polymorphic DNA) and ISSR (Inter-Simple Sequence Repeat) molecular markers were employed to determine the genetic homogeneity amongst the plantlets. Twelve (12) RAPD and nine (9) ISSR primers developed a total of 104 and 91 scorable bands, respectively. The band profiles of micropropagated plantlets were monomorphic to the mother, donor in vivo plant, and similarity values varied from 0.9542–1.000. The dendrogram generated through UPGMA (unweighted pair group method with arithmetic mean) showed 99% similarities amongst all tested plants confirming the genetic uniformity of in vitro raised plants.

RAPD primer screening for markers development of Jinten Hitam (Nigella sativa L.) authentication

Jinten hitam (Nigella sativa) is one of the medicinal plants in the family Ranunculaceae, native to the Mediterranean region and several regions of Asia. The seed of Nigella sativa is widely cultivated, distributed, and used in pharmaceuticals, cosmetics, and food industries. Due to similar seed morphology to its potential adulterant such as Corchorus spp., N. sativa seeds are susceptible to adulteration and substitution in markets. Molecular markers have become one of the most reliable methods for the identification and authentication of medicinal plants. The objective of this study was to select the random amplified polymorphic DNA (RAPD) primer for generating authentication methods of Jinten hitam (N. sativa). Genomic DNA was extracted from samples of N. sativa seed, Corchorus sp. seed, and a mixture of both samples. Forty-two random RAPD primers were used in this study. A total of 227 DNA fragments were produced from 37 amplified RAPD primers, out of which 65% were polymorphic. Primer OPK-4 and OPC-12 generated specific fragments in N. sativa, meanwhile, Primer OPB-1, OPL-5, OPM-3, OPD-5, and OPC-12 generated specific fragments for Corchorus. RAPD molecular marker was able to authentication Jinten hitam (N. sativa) and Corchorus sp. using a selected primer. This research was the first report on RAPD primer screening for Nigella sativa authentication from its potential adulterant (Corchorus spp.).

RAPD-based evaluation of genetic diversity among populations of the Iranian endemic species Rhabdosciadium aucheri Boiss. (Apiaceae).

Rhabosciadium aucheri is an Iranian endemic herbaceous species that grows in the west, center, and south regions of Iran. In the present study, genetic variation of 70 individuals belonging to seven natural populations of four provinces was investigated using Random Amplified Polymorphic DNA (RAPD) markers. Ten out of twenty-two RAPD primers employed in this study, generated 110 highly amplified and reproducible loci and a mean of 11.1 bands per primer and 48.13% of polymorphism was obtained. According to our results, the primer OPA10 presented the highest effective number of alleles, Shannon's index, and genetic diversity. The highest value of genetic identity (0.916) was determined between Hamadan, Nahavand and Hamadan, Alvand Mts. populations and the highest genetic distance (0.277) was observed between Hamadan, Asadabad and Kurdistan, Qorveh populations. Therefore, there is an obvious correlation between genetic diversity and geographical distribution. PCA was obtained based on RAPD molecular data and Neighbor Joining (NJ) dendrogram was provided successively. Similar results were attained employing UPGMA and Neighbor Joining dendrograms, supported by PCA ordination plot. Overall, almost moderate level of polymorphism was obtained by RAPD molecular markers at the population level that shows a reasonable amount of intraspecific variability. The reason might be due to the low level of gene flow between populations that could give rise to high genetic differentiation.

Identification of molecular marker associated with mungbean yellow mosaic virus resistance in mungbean [Vigna radiata (L.) Wilczek

Mungbean [Vigna radiata (L.) Wilczek] is an important legume which can be grown in varying environmental conditions, during all three crop seasons viz., kharif, rabi and spring/summer in India, as sole or inter crop for grain and green manure. It is an excellent source of easily digestible proteins with low flatulence, which complements the staple rice diet in Asia. Mungbean Yellow Mosaic Disease (MYMD) caused by whitefly (Bemisia tabaci) transmitted mungbean Yellow Mosaic Virus (MYMV) is an important constraint of mungbean. To fulfill future demands, there is a need to use molecular marker technology and other biotechnological interventions. Hence, Bulked Segregant Analysis (BSA) with a Random Amplified Polymorphic DNA (RAPD) marker technique was used to analyze the F2:3 individuals of Pusa Ratna (susceptible) × Meha (resistant) to identify the molecular marker linked to MYMV resistance in mungbean. Field screening of thirty-five mungbean genotypes against MYMV was carried out, prior to crossing program to identify the resistant and susceptible genotypes. The RAPD primer, OPP 07 showed the specific band of 895 bp in resistant parent and their bulks, but not in the susceptible parent and their bulks. Co-segregation analysis was performed in resistant and susceptible F2:3 individuals; it confirmed that OPP 07895 marker was associated with MYMV resistance in mungbean. The linked RAPD molecular marker OPP 07895, can be used for the identification of (Quantitative Trait Locus) QTL for MYMV resistance and marker assisted selection program in mungbean.

Genetic variations of strawberry cultivars of Fragaria x ananassa and Fragaria vesca based on RAPD

Abstract. Aristya GR, Kasiamdari RS, Setyoningrum R, Larasati B. 2019. Genetic variations of strawberry cultivars of Fragaria x ananassa and Fragaria vesca based on RAPD. Biodiversitas 20: 770-775. In Indonesia, the increasing market demand for strawberries (Fragaria spp.) is not comparable to increased strawberry productivity. One of the efforts made to increase strawberry productivity with superior quality is plant breeding. The purpose of this research was to determine the genetic variation, lineage, and similarity index in some strawberry cultivars using molecular markers of Random Amplified Polymorphic DNA (RAPD). Eleven strawberry cultivar samples were taken from Indonesian Citrus and Subtropical Fruits Research Institute (Balitjestro), Batu City, East Java, Indonesia and Strawberry Agritourism in Banyuroto Village, Magelang District, Central Java, Indonesia. DNA isolation using modified CTAB buffer method. DNA amplification using PCR-RAPD method with 5 primers, namely UBC-516, UBC-594, OPA 10, OPA 16, and OPG 11. Strawberry lineage dendrogram construction was analyzed with clustering of Unweight Pair-Group Using Arithmetic Average (UPGMA) software Multi-Variate Statistical Average (MVSP). The research results showed that the 5 RAPD primers used in 11 strawberry cultivars produced 30 polymorphic DNA bands and 20 monomorphic DNA bands so it can be concluded that the genetic variation among 11 strawberry cultivars can be detected using RAPD molecular markers. The lineage of 11 strawberry cultivars that have the highest similarity index is found in Earlibrite and Rosalinda II cultivars of 98.85%.

English

To assay the efficiency of graphite nanoparticles (GtNPs) in sterilizing tissues and their role in enhancing genetic diversity, sweet potato is considered an important crop; hence its explants were used. In this experiment, GtNPs of 200, 400 and 800 ppm concentrations were used for sterilization of MS callus induction and regeneration media in Abees cultivar. The results showed that GtNPs had a good potential for removing bacterial contaminants without having side effects on the explant viability during the sterilization of sweet potato tissue in all their concentrations. Also, the percentage of callus induction increased from 98.67% in control to 100% in all GtNPs concentrations. The number of shoots per callus was enhanced at 400 ppm concentration. RAPD molecular markers and SDS-PAGE analysis were used to assess the genetic diversity of the sweet potato selected plants obtained from somaclonal variations in combination with GtNPs. Five decamer random amplified polymorphic DNA (RAPD) primers generated a total of 96 DNA fragments from the selected variants and their parent. Out of them, 82 polymorphic bands appeared with 85.42% polymorphism. The levels of DNA and protein patterns polymorphism within each treatment varied. RAPD and protein markers revealed that the concentration of 800 ppm showed the lowest similarity average among the ten selected variants and their parent. The obtained results indicated that somaclonal variation with GtNPs can be combined to increase the induced mutations frequency. Key words: Graphite nanoparticles, somaclonal variation, random amplified polymorphic DNA (RAPD), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).

Species delimitation in the genus Tamarix: Morphological and molecular data<br />

Tamarix (Tamaricaceae) is an ancient genus in Asia that contains about 54 to 90 species with unresolved taxonomy. Tamarix has major center of speciation in the Pakistan-Afghanistan-Iran-Turkmenistan-S-Kazakhstan-W-China area and in the eastern Mediterranean area. Tamarix species are highly plastic and tolerant to adversity being able to occupy different habitats, thus showing a wide range of ecological variations. Tamarix species have commercial value in the landscape and horticultural trade and are planted for erosion control and as wind breaks. In the genus Tamarix, many species are morphologically very similar probably due, in part, inter-specific hybridization. Tamarix species live in sympatry in Sistan and Baloochestan province of Iran and we have no report on the number of species and their genetic structure. Therefore, we performed morphometric study to delimit the species growing in these two provinces and also used combined ISSR (Inter simple sequence repeat) and RAPD (Random amplified polymorphic DNA) molecular markers to study their genetic structure. Morphological analyses separated plants of 6 studied species in different groups, indicating that morphological characters used can delimit Tamarix species. We also encountered some plants having intermediate morphological characters. AMOVA and Hickory test showed significant genetic differences of the studied species and Mantel test showed significant correlation between genetic distance and geographical distance. STRUCTURE analysis and population assignment test revealed some degree of genetic admixture among the studied species. Assignment tests showed that gene flow mainly occurred among the species that were growing in neighborhood regions. This genetic admixture may be the reason for intermediate morphological characters observed in some the plants.

Phylogenetic analysis among some pome fruit trees of Rosaceae family using RAPD markers

ABSTRACTTo evaluate the phylogenetic relationships among subtribe Pyrinae of Rosaceae, 50 different genotypes and cultivars of pome fruit trees were collected from various locations in Iran, and analysed using random amplified polymorphic DNA (RAPD) markers. Altogether, 85 polymorphic fragments were produced by 11 random 10-mer primers. The number of produced bands varied from 4 to 12 for each primer, 7.72 on average. The lowest Jaccard's genetic similarity coefficient was scored between apple cv. ‘Akan’ and a wild Pyrus syriaca pear (0.04), and the highest similarity was observed between two genotypes of P. syriaca (0.97). Cluster analysis using the unweighted pair group method with arithmetic mean (UPGMA) properly separated the accessions and divided them into three major groups, including Malus, Pyrus, Mespilus and Crataegus. Members of Mespilus and Crataegus were further separated and formed two subgroups. Analysis of the genetic structure, using STRUCTURE and phylogenetic relationship using TreeView software confirmed the results of the UPGMA dendrogram and properly distinguished different genera. Among different genera studied in our experiment, Malus was the most distinct one and showed high genetic distance from other genera. Generally, the results from the present study provide valuable information about the genetic relationship among important pome fruit trees in Iran and indicate that, although Mespilus is genetically close to the Crataegus genus, it has relatively high similarity with Pyrus and Malus. Moreover, outcomes of this research confirmed that RAPD molecular markers are an appropriate means for evaluation of the genetic relationship among various genera of pome fruit trees.

Genetic and cytological diversity in cherry tree accessions ( Eugenia involucrata DC) in Rio Grande do Sul

This study aimed to evaluate the genetic and cytological diversity and stability of 35 cherry tree accessions collected in Rio Grande do Sul. We used 15 RAPD (Random Amplified Polymorphic DNA) molecular markers and performed cytological analysis and number count of anthers. Analyses of genetic diversity allowed the separation of accessions into four groups, resulting in an average of 8.93 bands per primer amplified, 7.89 polymorphic bands, 88.08% of polymorphism and 86% of genetic similarity. Cytological analyses of gametic cells allowed for the characterization of accessions as diploids with n=11. In these, the average of meiotic cells considered normal was 82.12%; average pollen viability was 92.44% and in vitro germination was 40.26%; the average number of anthers was 161.85 anthers/flowers. Therefore, the accessions evaluated showed high genetic similarity and cytological stability and can be used in commercial plantations or hybridizations.

Genetic Diversity of Peanut (Arachis hypogea L.) Cultivars as Revealed by RAPD Markers

The objectives of this study were to evaluate the genetic diversity of the peanut accessions using Random Amplified Polymorphic DNA (RAPD) molecular marker and to evaluate RAPD markers to be used in peanut as genetic markers and improve such techniques as suitable strategies for peanut germplasm characterization. Twenty peanut accessions were included in this study and were subjected to RAPD molecular markers analysis. Twenty-seven RAPD primers produced 210 amplification products of which 80 (36.4%) were polymorphic. In conclusion, this study reported a successful fingerprinting of peanut accessions using RAPD markers and demonstrated the usefulness of these markers in estimating the extent of genetic variation in peanut germplasm.

Density-weighted Algorithms for Similarity Computation and Cluster Tree Construction in the RAPD Analysis of Natural Cordyceps sinensis

Objective: The goal of this study was to develop and validate density-weighted arithmetic methods for the analysis of the dynamic maturational changes in random amplified polymorphic DNA (RAPD) polymorphisms in Cordyceps sinensis containing multiple fungi. Methods: Ten random primers were used for PCR amplification to monitor changes in the RAPD molecular marker polymorphisms in caterpillar body, stroma and ascocarp portion samples of C. sinensis collected at different stages of maturation. We compared (1) the density-unweighted Nei-Li similarity equation and new similarity equations considering the densities of DNA bands and (2) the density-unweighted and weighted arithmetic methods for cluster analysis. Results: The algorithm using the Nei-Li similarity equation did not account for the differences in the density of the matched randomly amplified amplicons, whereas the new similarity equations were capable of integrating different amplicon densities into the similarity computation. With improvements in the similarity computation and cluster construction, the new methods revealed dynamic maturational changes in the RAPD polymorphisms of molecular markers in C. sinensis caterpillar bodies and stromata. The polymorphism analysis revealed similarities of 74%-88% between the RAPD polymorphisms of the ascocarp portion versus premature and mature C. sinensis stromata, but low similarities of<70% versus the caterpillar bodies and maturing stromata. Apparent dissimilarities (similarity <66%) were also found between Hirsutella sinensis and C. sinensis samples. Conclusions: These new algorithmic methods represent an advance in similarity computations when comparing polymorphisms of RAPD molecular markers in C. sinensis and provide accurate analytic tools to capture the molecular information related to the dynamic

Heterogeneity in the Second Internal Transcribed Spacer (ITS2) of rDNA in the Genetically Distinct Metopolophium dirhodum Isofemale Lineages (Hemiptera: Aphididae)

The aim of this study was to investigate the level of evolutionary divergence among five isofemale lineages of Metopolophium dirhodum Walker (Hemiptera: Aphididae) using sequences of the Internal Transcribed Spacer (ITS) of ribosomal DNA (rDNA). These isofemalelineages were derived from parthenogenetic females, previously genetically characterized by having strong association of RAPD (Random Amplified Polymorphic DNA) molecular markers and host plant preference. Out of 25 sequences, only 17 containing ITS2 (the second spacer) were used for analysis due to their quality. Within 250 nucleotide sites originated by multiple alignments, 10.4% were variable, with the predominance of indels over substitutions. The average of nucleotide diversity (π) within isofemale lineages was similar to the diversity found among all isofemale lineages. The topology inferred by the Neighbor-Joining method showed that no evolutionary divergence inferred by ITS2 sequences among isofemale lineages typified by RAPD could be associated with host preference. The ITS2 sequence differences found within lineages and their causes are discussed.

Genetic relationship among Camponotus rufipes Fabricius (Hymenoptera: Formicidae) nests by RAPD molecular markers

Random amplified polymorphic DNA (RAPD) markers were used to investigate the genetic relationship among nests of the carpenter ant, Camponotus rufipes, located in the same area. Five random oligodecamers were used to amplify DNA from 108 ant workers collected from six nests. A total of 47 RAPD markers were identified, which revealed low levels of genetic differentiation among nests (Φst = 0.00218) and a low average Shannon index (0.3727) among workers within nests. These results together suggest that the C. rufipes nest may be formed by a single, once-mated queen and that nests produced by queens that are genetically related tend to keep their nests in close proximity to one other.

The Use of ISSR and RAPD Markers for Genetic Diversity among South Tunisian Barley

Random amplified polymorphic DNA (RAPD) and intersimple sequence repeat (ISSR) were assayed to determine the genetic diversity of 80 barley specimens from South Tunisia. The ISSR primers showed variation in the percentage of polymorphism, band informativeness (Ib), and resolving power (Rp). The percentage of polymorphism is 66.67%, the average Ib ranged from 0.24 to 0.39, while Rp ranged from 0.74 to 1.16. In RAPD analysis, three primers yielded a total of 17 scorable bands, which are all polymorphic. The three polymorphic primers exhibited variation with regard to average band informativeness (AvIb) and resolving power (Rp). RAPD and ISSR marker systems were found to be useful for the genetic diversity among the barley specimens. The two dendrograms obtained through these markers show different clustering of 80 barely specimens, but we noted that some clusters were similar in some cases. A poor correlation () was found between both sets of genetic similarity data, suggesting that both sets of markers revealed unrelated estimates of genetic relationships. Therefore, the ISSR and RAPD molecular markers show two genetic grouping of studied barely specimens.

Variabilidade genética de acessos de pitaya com diferentes níveis de produção por meio de marcadores RAPD

A espécie de pitaya mais cultivada atualmente é Hylocereus undatus, a pitaya-vermelha-de-polpa-branca. Colômbia e México são os principais produtores mundiais e, devido à sua rusticidade, a pitaya é considerada uma alternativa potencialmente viável também para o aproveitamento de solos pedregosos, arenosos e maciços rochosos. Apesar da crescente demanda, ainda não há uma cultivar lançada no mercado que atenda às necessidades climáticas de produção e às exigências do consumidor brasileiro. O presente trabalho é parte do programa de seleção e melhoramento da pitaya CPAC PY-01 da Embrapa Cerrados. Objetivou-se realizar o estudo da variabilidade genética de 16 acessos de pitayas mantidos na coleção de germoplasma da Embrapa Cerrados, apresentando diferentes características fenotípicas relacionadas especialmente à produção, por meio de marcadores moleculares RAPD (Random Amplified Polymorphic DNA). O DNA genômico de cada acesso foi extraído, e onze iniciadores decâmeros foram utilizados para a obtenção de marcadores moleculares RAPD, que foram convertidos em matriz de dados binários, a partir da qual foram estimadas as distâncias genéticas entre os acessos e realizadas análises de agrupamento e de dispersão gráfica. Foram obtidos 111 marcadores RAPD, perfazendo uma média de 10,1 marcadores por primer, dos quais 45 (40,54%) foram polimórficos. As distâncias genéticas entre os 16 acessos variaram entre 0,006 e 0,148. As maiores distâncias genéticas foram obtidas entre os acessos "52" e "61", sendo que, em 2007, o primeiro produziu mais de 25 frutos, e o segundo, nenhum. Assim, deduz-se que, nesse caso, a próvável causa da variação seja genotípica. As menores distâncias genéticas foram constatadas entre os acessos "63"e "55" e entre "19"e "59". Os dois grupos apresentaram valores de produção próximos. Os marcadores moleculares RAPD mostraram que, mesmo dentro da mesma espécie, há variabilidade genética entre plantas com produções diferentes, ressaltando a importância das técnicas moleculares para subsidiar e auxiliar nos trabalhos de seleção, em programas de melhoramento genético.

KERAGAMAN GENETIK POPULASI Araucaria cunninghamii MENGGUNAKAN PENANDA RAPD (RANDOM AMPLIFIED POLYMORPHIC DNA)

Genetic diversity was needed for supporting genetic conservation and tree improvement program. Araucaria cunnighamii is one of Indonesian conifer species which has various potential uses. Information about genetic diversity and its distribution, and genetic relationship can be revealed using RAPD (Random Amplified Polymorphic DNA) molecular marker. The objective of the study was to analyze genetic diversity within and among 8 populations of A. Cunninghamii, and genetic relationship between populations. Leaf samples of 64 trees were collected from 7 population in Papua and 1 population in Queensland (Australia). Genetic diversity was analyzed using 23 RAPD primers with 68 polymorphic loci. Mean genetic diversity within population was 0.270, and mean genetic distance among populations was 0.092. Some of the samples from the same population were grouped in the same cluster. But generally, the clustering of the samples. Genetic relationship among A. cunninghamii populations revealed that 8 populations were divided into two groups. First group was consisted of 7 population in Papua (Indonesia) and then split into 5 subcluster. Second group was population in Queensland (Australia).

English

&nbsp; With orthogonal analysis by L27(36), the random amplified polymorphic DNA (RAPD)-PCR optimization reaction system for camellia were obtained. Results showed that the optimization system was 10&times;PCR Buffer (2.5 &mu;L), 25 mM MgCl2(2.5 &mu;L), 2.5 mM dNTPs (2.0 &mu;L), 20 &mu;M primer (1.0 &mu;L), Tag (1.5 U), temple DNA (40 ng or so) and added ddH2O to the total volume 25 uL; suitable annealing temperature was 36&deg;C.&nbsp;With the optimized system and fifteen 10 nt random primers, we analyzed 15 camellia cultivars and observed 102 clear amplified loci, in which polymorphic loci were 79 while the percentage of polymorphic loci were 77.54%. Cluster analysis showed that the four groups were divided at the point 0.75 of similarity coefficient,&nbsp;indicating relatively high genetic diversity. We also found that the gene controlling petal color may play an important role in RAPD analysis. Moreover, genetic diversities based on RAPD analysis could be clearly reflected by morphological traits among 15 camellia cultivars. This study showed the RAPD optimization system was suitable and RAPD molecular marker was effective and useful tool for detection of genetic relationships among camellia cultivars. &nbsp; Key words:&nbsp;Optimization, RAPD, Camellia cultivars, genetic relationships.