Random Amplified Polymorphic DNA Banding Patterns Research Articles

Zinc oxide nanocatalyst mediates cadmium and lead toxicity tolerance mechanism by differential regulation of photosynthetic machinery and antioxidant enzymes level in cotton seedlings

Cadmium (Cd) and Lead (Pb) heavy metal pollution induced toxicity severely affects the plant growth and yield of various agriculutral crops worldwide. The present study discuss the prime role of phycomolecules coated zinc oxide nanoparticles (ZnONPs) application on development of heavy metal tolerance mechanism in cotton (Gossypium hirsutum L.) seedlings better than exposed to Cd and Pb treatments alone. Co-exposure of ZnONPs along with heavy metal treatments significantly promoted the shoot, and root growth as well as biomass compared to control, while it was down-regulated in Cd and Pb exposed seedlings. The intervention of ZnONPs had up-regulated the level of chlorophyll a, b and carotenoid contents in leaves grown under Cd and Pb treatments than the untreated control. Similarly, the level of total soluble protein and malondialdehyde (MDA-lipid peroxidation) contents was significantly increased in the co-presence of ZnONPs along with Cd and Pb treatments over their respective control. Accumulation of antioxidant defense enzymes viz., superoxide dismutase (SOD), catalase (CAT), peroxidase (POX) and ascorbate peroxidase (APX) was up-regulated significantly in seedlings upon co-exposure of ZnONPs with Cd and Pb treatments. Random amplified polymorphic DNA (RAPD) fingerprinting analysis exhibited no genomic changes/alterations in seedlings by co-existence of ZnONPs with heavy metals. Overall, the present results indicate that the addition of ZnONPs with Cd and Pb ion exposure protects cotton seedlings by alleviating heavy metal induced phytotoxicity and promoted physiochemical characteristics via differential regulation of photosynthetic machinery as well as antioxidative defense mechanisms in cotton seedlings. Results strongly suggest that phycomolecule coated ZnO nanoparticles could be effectively used as nanofertilizer to cultivate agronomically important crops in heavy metal polluted soil in the future.

Genetic Diversity of Artemisia herba-alba in Libyan Green Mountain

The genetic variability of 18 individuals of Artemisia herba-alba from 3 different elevations in Green Mountain—Libya was examined using sequence-related amplified polymorphism (SRAP) and random amplified polymorphic DNA (RAPD) techniques. Results showed distinctive SRAP and RAPD banding pattern in this species scoring a total of 17 SRAP and 61 RAPD bands including 94.12% and 100% polymorphic with amplicon size ranging from 50 to 1600 bp and 150 to 1800 bp, respectively. Phylogenetic relationship between the genotypes was able to divide the samples into two main groups with genetic similarity value ranged from 41 to 84%, indicating that the genetic relationship is slightly related to the geographical location of the studied plant area.

Evidence of persistence of Prevotella spp. in the cystic fibrosis lung.

Purpose. Prevotella spp. represent a diverse genus of bacteria, frequently identified by both culture and molecular methods in the lungs of patients with chronic respiratory infection. However, their role in the pathogenesis of chronic lung infection is unclear; therefore, a more complete understanding of their molecular epidemiology is required.Methodology. Pulsed Field Gel Electrophoresis (PFGE) and Random Amplified Polymorphic DNA (RAPD) assays were developed and used to determine the degree of similarity between sequential isolates (n=42) from cystic fibrosis (CF) patients during periods of clinical stability and exacerbation.Results. A wide diversity of PFGE and RAPD banding patterns were observed, demonstrating considerable within-genus heterogeneity. In 8/12 (66.7 %) cases, where the same species was identified at sequential time points, pre- and post-antibiotic treatment of an exacerbation, PFGE/RAPD profiles were highly similar or identical. Congruence was observed between PFGE and RAPD (adjusted Rand coefficient, 0.200; adjusted Wallace RAPD->PFGE 0.459, PFGE->RAPD 0.128). Furthermore, some isolates could not be adequately assigned a species name on the basis of 16S rRNA analysis: these isolates had identical PFGE/RAPD profiles to Prevotella histicola.Conclusion. The similarity in PFGE and RAPD banding patterns observed in sequential CF Prevotella isolates may be indicative of the persistence of this genus in the CF lung. Further work is required to determine the clinical significance of this finding, and to more accurately distinguish differences in pathogenicity between species.

Identification and Differentiation of Egyptian Isolates of Erwinia amylovora using 16S rRNA Gene and RAPD-PCR

Erwinia amylovora the causative of fire blight disease on pear in Egypt was identified with some classical tests and molecular methods. Random Amplified Polymorphic DNA (RAPD) technique was used to identify the genetic variability among 6 Egyptian isolates of E. amylovora isolated from different governorates (Behera, Gharbiya, Nubariya, Menufiya, Kaluobiya and Ismailiya) and compared with one reference isolate. Profiles were obtained by amplification of DNA from these isolates with six random primers.The RAPD band patterns resulting from these primers were analyzed using UPGMA method to construct a similarity matrix and generate a dendogram indicating the relationships between the examinedisolates. The results obtained indicated that primers P4d, P5e and P6f were chosen because they reliably and reproducibly detected polymorphism among the selected isolates. Also, no variation in bacterial genome was observed among the examined isolates. In addition, 16SrRNA gene sequence was used to identify E. amylovora isolates using universal primers (8F and U1492R). The results showed that sequence of the isolates collected from different locations was matched with the sequence found in the Genbank database. The sequence of the fragment (16S rRNA gene) showed percentage homology with strain DSM30165 of Genbank strains ranged from 89–97%. The selected isolates were found to have theclosest homology to E. amylovora.

Lack of evidence for sexual transmission of genital Candida species among women who have sex with women: a mixed methods study

ObjectiveThe contribution of sexual transmission to genital Candida infection remains unclear. This study sought to investigate whether sexual behaviours were associated with the presence of genital Candida species among a...

Relationships between Malaysians Cultivars of Tongkat Ali (Eurycoma longifolia Jack) Obtained through RAPD Analysis

Random Amplified Polymorphic DNA (RAPD) analysis was done on Eurycoma Longifolia Jack (ELJ) cultivars originating from 7 different locations in Malaysia to determine whether the originality of ELJ cultivars can be preserved in an uncontrolled cultivation area. Out of 60 arbitrary 10-mer primers, a total of 320 DNA fragments were amplified with an average of 53.3 RAPD markers per primer. 71 (22%) of the 320 DNA-fragments was found to be monomorphic. The remaining 249 (78%) were polymorphic. The approximate size of the largest fragment produced was 3000 base pair (bp) and the easily recognizable fragment produced was 300 bp. Similarity index of RAPD banding patterns was used to investigate the relationships between ELJ cultivars. Mantel test showed that the correlation between Jaccard and Dice similarity matrices was high and significant (0.9907). The dendogram showed that ELJ cultivars can be clustered into three main clusters. Cluster 1 consisted of Sabah cultivars (East Malaysia), Cluster 2 consisted of Pahang and Terengganu cultivars (middle part of West Malaysia) while cluster 3 consisted of Kedah and Kelantan cultivars (east part of West Malaysia). The results of Principle Coordinates Analysis (PCA) were comparable to the Dendogram by UPGMA. The first two principle coordinates explained 53.62% of total variations and separated the 37 ELJ cultivars into three main clusters with a slight overlap between each other.

Quality Parameters and RAPD‐PCR Differentiation of Commercial Baker's Yeast and Hybrid Strains

Baker's yeast, Saccharomyces cerevisiae, is a key component in bread baking. Total of 12 commercial baker's yeast and 2 hybrid strains were compared using traditional quality parameters. Total of 5 strains with high leavening power and the 2 hybrid strains were selected and evaluated for their alpha-amylase, maltase, glucoamylase enzymes, and compared using random amplified polymorphic DNA (RAPD). The results revealed that all selected yeast strains have a low level of alpha-amylase and a high level of maltase and glucoamylase enzymes. Meanwhile, the Egyptian yeast strain (EY) had the highest content of alpha-amylase and maltase enzymes followed by the hybrid YH strain. The EY and YH strains have the highest content of glucoamylase enzyme almost with the same level. The RAPD banding patterns showed a wide variation among commercial yeast and hybrid strains. The closely related Egyptian yeast strains (EY and AL) demonstrated close similarity of their genotypes. The 2 hybrid strains were clustered to Turkish and European strains in 1 group. The authors conclude that the identification of strains and hybrids using RAPD technique was useful in determining their genetic relationship. These results can be useful not only for the basic research, but also for the quality control in baking factories.

A Set of SCAR Markers Efficiently Differentiating Hybrid Rice

Molecular markers have been widely used in crop genetic improvement, seed test and genetic mapping. Of which, sequence characterized amplified region (SCAR) markers are particularly popular for its diversity, stable reproducibility, and suitability for analyzing large number of samples. In this study, 500 random amplified polymorphic DNA (RAPD) primers were tested, and a set of SCAR markers comprising 37 pairs of loci-specific primers were developed from the DNA fragments ranging from 300 to 1000 bp which correspond to the stable, distinctive RAPD banding patterns. Using these SCAR markers, 59 hybrid rice combinations were assessed and distinguished into 58 subgroups at the similarity coefficient of 0.97 in a genetic clustering tree based on the allele diversities of the SCAR markers. Furthermore, 13 hybrid rice combinations were reassayed with 40 randomly selected simple sequence repeat (SSR) markers to evaluate the effectiveness of these SCAR markers. SSR markers produced similar results to SCAR markers as the 13 hybrid rice combinations were completely separated at the similarity coefficient of 0.91 in the clustering tree established from SSR patterns. Taken together, SCAR markers prove to be effective tools for identifying and differentiating hybrid rice combinations.

Genetic diversity and relationships detected by inter simple sequence repeat (ISSR) and randomly amplified polymorphic DNA (RAPD) analysis among Polygonum species growing in North of Iran

The phylogenic relationship within four species of Polygonum (including Polygonum hyrcanicum Rech. f. (three samples), Polygonum persicaria Boiss & Bushe Boiss, Polygonum avicular L., and Polygonum hydropiper L. subsp. Hydropiper ) was investigated by randomly amplified polymorphic DNA analysis (RAPD) and inter simple sequence repeat (ISSR) markers. All the species were clearly identified using 20 RAPD primers (10-mer ZO1- ZO20) and 15 ISSR primers (S1-S15). The total number of amplification products produced with 20 primers of both markers was 315. Genetic distance was calculated in order to construct phylogenic dendrogram or phylogram of closely related samples. Results indicate that the genetic distance between two samples of P. hyrcanicum originated from Sari and Goharbaran was considered to be short (0.622) and their RAPD and ISSR banding patterns were quite similar to each other also there is a close relationship between these two samples of Polygonum with P. avicular (0.848). The species P. hydropiper is not only far from two samples of P. hyrcanicum but also, has different patterns of RAPD and especially ISSR profiles compared to other samples. It is also interesting that P. hyrcanicum , gathered from Veresk, has no close relationship with other pairs of P. hyrcanicum . More chemotaxonomic investigation is needed to support the phylogenetic classification trees. Key words : Polygonum, Polygonaceae, molecular markers, P. hyrcanicum, P. persicaria, P. avicular, P.hydropiper

Prevalence, genetic diversity, and antibiotic susceptibility of Cronobacter spp. (Enterobacter sakazakii) isolated from Sunshik, its ingredients and soils

The prevalence, genetic diversity, and antibiotic susceptibility of Cronobacter species (Enterobacter sakazakii) isolated from sunshik products, its ingredients, and root vegetable farm’s soils were investigated to analyze main reservoirs and contaminated sources of Cronobacter spp. Cronobacter spp. was isolated from 9 of 15 sunshik products, 26 of 72 its ingredients, and 2 of 39 soils. The root vegetables such as sweet potato and carrot showed higher contamination rate (70%) than the other sunshik ingredients. All isolates showed 929 bp band amplified from 16S rRNA and resistant to ampicillin, amoxicillin/clavulanic acid, and cefazolin. All isolates showed diverse random-amplified polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) band patterns. However, 3 cases of RAPD banding patterns between clinical strains and isolates from sunshik products and root vegetables (yam, carrot) were related with similarities level of 80%. These studies indicated that root vegetables can be an important contamination source of Cronobacter spp. in sunshik products. Thus, the preparation of root vegetables for manufacturing sunshik products used as a weaning diet was handled with more care than the other sunshik ingredients.

Typing of Brachyspira spp. from rodents, pigs and chickens on Swedish farms

The aim of the current study was to look for evidence of possible cross-species transmission of Brachyspira species between rodents and farm animals. To do this, previously collected and characterised Brachyspira isolates from rodents, pigs and chickens on the same farms were analysed by random amplified polymorphic DNA (RAPD). Isolates with similar RAPD banding patterns were further typed by pulsed-field gel electrophoresis (PFGE). Identical isolates of Brachyspira pilosicoli, Brachyspira intermedia, Brachyspira murdochii and Brachyspira innocens from pigs and rodents and of B. murdochii from laying hens and rodents were found, indicating cross-species transmission at farm level. PFGE data from rodent isolates of Brachyspira hyodysenteriae were compared with PFGE data from previously typed field isolates of B. hyodysenteriae from pigs with swine dysentery and isolates from mallards ( Anas platyrhynchos). Three of four isolates of B. hyodysenteriae from rodents were similar to porcine field isolates by PFGE. PCR analyses of the plasmid-encoded and potential virulence determinants rfb genes B, A, D and C showed that they were present in isolates of B. hyodysenteriae of porcine, mallard and rodent origin.

English

  Eight isolates of Botrytis cinerea, causal organism of chickpea from eight different locations of western and eastern Indo-Gangetic Plains (IGP) of India were analyzed for morphological, cultural, virulence and genetic variations.  Characterization of virulence and genetic variations of the isolates was based on their pathogenicity against 40 selected chickpea genotypes and randomly amplified polymorphic DNA (RAPD) analysis, respectively. The isolates differed in their optimum growth, temperature, conidial formation and size of conidia on potato dextrose agar. Based on cluster analysis, the eight test isolates were separated into three pathotypes with two isolates from western and four from eastern IGP grouped together with >80% similarity. Based on cluster analysis of the RAPD banding patterns, genetic similarity of the isolates varied from 14-44%, and the isolates were separated into three groups. However, pathotypes variation detected among B. cinerea isolates could not be differentiated based on the RAPD markers examined.   Key words: Botryotinia fuckeliana, Botrytis gray mold, variability.

Genetic Diversity of Hevea IRRDB’81 Collection Assessed by RAPD Markers

The majority of Hevea (Hevea brasiliensis Muell. Arg.) genetic resource in Vietnam derived from the IRRDB'81 germplasm collected in the Amazonian habitats of the genus. Random amplified polymorphic DNA (RAPD) analysis was used to examine the genetic diversity and structure of the IRRDB'81 germplasm. A total of 59 accessions from 13 different districts of the Brazilian states namely Acre, Rondonia, and Mato Grosso were brought into the study using six arbitrarily preselected primers. Sixty-five RAPD band patterns ranging in size from 0.2 to 3.0 kbp were scored for analysis. Differences in the level of DNA polymorphism among the districts and states were revealed. The percentage of the polymorphic DNA fragments calculated for 13 individual districts varied from 15.38 to 70.77%. The mean values of heterozygosity within the district varied from 0.064 to 0.264. Pairwise district Nei's genetic distance values ranged from 0.046 for Catriquacu and Itanba of Mato Grosso to 0.304 for Tarauaca of Acre and Aracatuba of Mato Grosso. The estimated values of Shannon's diversity index ranged from 0.093 for the Assis-Brasil district of Acre to 0.389 for the Jiparana district of Rondonia. The analysis of molecular variance (AMOVA) indicated that most of the genetic variations were found among accessions within the districts, while interdistrict variance component accounted for 14.1% only. The low interdistrict differentiation probably implied an extensive gene flow among them. Both the principal coordinate analysis and UPGMA cluster analysis based on genetic distance values revealed a varying degree of separation among the districts and that conformed to geographical origins of Hevea IRRDB'81 collection.

RAPD differentiation of Malassezia spp. from cattle, dogs and humans

The molecular characterization of Malassezia spp. isolates from animals and humans has not been thoroughly studied. We have analysed the DNA profile by random amplified polymorphic DNA (RAPD)-PCR to compare the genetic diversity between isolates from the external ears of cattle, dogs and humans. The analysis of electrophoretic profiles on 8% polyacrylamide gel and their phenograms showed genetic heterogeneity between RAPD profiles of Malassezia furfur and Malassezia slooffiae isolates from humans and cattle and between Malassezia pachydermatis isolates from dogs and cattle. Intra-species variations in DNA pattern of Malassezia isolates and the presence of specific genetic types in cattle, dogs or humans were observed. A review of genetic heterogeneity of these yeast in veterinary and human medicine studies is given considering a possible transmission animal to human or human to animal. Additional studies must clarify the differences between the RAPD band patterns observed in this and other studies, which would facilitate monitoring of Malassezia spp. carriage in domestic animals and in humans.

Application of RAPD and SI-typing techniques to confirm genetic integrity of radish ( Raphanus sativus L.) varieties

The random amplified polymorphic DNA (RAPD) and self-incompatibility (SI) typing techniques were applied to settle a lawsuit between a Seed Company and a farmer concerning seed impurity in radish ( Raphanus sativus L.). After sowing “Halra Woldong” (HW) a winter radish variety, two morphologically different radish plants grew. Genomic DNA from each of the two types were prepared and analyzed using ninety-six RAPD markers and four variety-specific markers were found. The polymorphic RAPD band patterns were compared with those of the twenty Korean radish varieties. The genetic integrity of the two varieties in question was identified and further confirmed by SI-typing of S-locus glycoprotein ( SLG) and S-locus receptor kinase ( SRK) genes.

DNA fingerprinting of Fusarium solani isolates related to a cutaneous infection in a sea turtle

Random amplified polymorphic DNA (RAPD) fingerprinting was applied to Fusarium solani from a cutaneous hyalohyphomycosis in a loggerhead sea turtle. A total of seven F. solani isolates were examined, three from culture collections and four from the turtle infection (one from the turtle's lesions and three from the sand of the tank in which the turtle was kept). The banding patterns of the isolates from culture collections were markedly different from the patterns of the isolates from both the turtle and the sand. The RAPD banding patterns were the same with all the primers used, suggesting that this opportunistic infection may be related to the presence of F. solani in the tank. RAPD techniques can be particularly useful for large-scale epidemiological studies and for identifying sources of infection.

First Report of Race 3 of Bipolaris zeicola on Corn in China.

In Meixian County of Shaanxi Province, China, during the summer of 2002, mature corn plants in a field plot showed severe leaf spot symptoms. The lesions were narrow (3.5 to 18 mm long and 0.4 to 1.5 mm wide), grayish tan, and surrounded by a light- to dark-pigmented border. Leaves wilted when lesions coalesced. From 2002 to 2005, the disease was observed in other Shaanxi Province counties, including Yangling, Wugong, Qianxian, Longxian, and Qianyang, although in most cases, symptom development was less severe than it was in Meixian. Seven isolates from four counties were obtained by isolation from host tissue on potato dextrose agar (PDA), followed by single-spore culturing and incubation on PDA at 25°C in the dark for 7 days. Conidial suspensions were prepared from a single-spored culture on PDA plates. Pathogenicity tests were performed by spraying five corn seedlings (cv. Yuyu 22) at the three- to four-leaf stage in separate 10-cm-diameter pots with 10 ml of a conidial suspension (106 spores per ml) per plant. Each of three isolates was used in separate inoculations that were performed in different weeks. Controls were sprayed with sterile distilled water only. Plants were covered with plastic bags for 48 h and incubated at 23 to 25°C in a chamber. One week after inoculation, leaves on all inoculated plants developed characteristic lesions, whereas untreated controls had no symptoms. The pathogen was reisolated from diseased leaves on PDA after surface sterilization with 2% NaOCl. On PDA, proliferation of conidia usually occurred on all sides of the conidiophore. Conidiophores were cylindrical, simple, smooth, septate, and straight to flexuous. Conidia were 49 to 89 μm long and 11 to 17 μm wide, with 3 to 10 distosepta, straight or moderately curved, dark or olivaceous brown, and the cells on the ends sometimes appeared paler than those in the middle. These characteristics match those of Bipolaris zeicola (Stout) Shoemaker. On the basis of the arbitrary primers selected by Jones et al. (1), random amplified polymorphic DNA (RAPD) analysis was used for species and physiological race determination. A single DNA fragment approximately 1.2 kb, which is characteristic of B. zeicola, was amplified from all seven isolates with arbitrary primer A20 (5'CTTGGATTC3'). Analysis of PCR products obtained with arbitrary primer A03 (5'AGTCAGCCAC3') showed that all seven isolates lacked 2,700- and 2,300-base bands, and therefore, sorted into B. zeicola race 3. On the basis of pathogenicity, morphology, and RAPD band patterns of primer A20, the fungus was confirmed as B. zeicola. The shape of leaf lesions and RAPD band patterns using primer A03 showed further that the pathogen was race 3 of B. zeicola. Bai et al. (2) reported race 1 and race 2 of B. zeicola in China, but to our knowledge, this is the first report of race 3 in China.

Evidences on the migratory movements of the nectar-feeding bat Leptonycteris curasoae in Mexico using random amplified polymorphic DNA (RAPD)

We examined the genetic relatedness of six populations of Leptonycteris curasoae in Mexico using random amplified polymorphic DNA (RAPD). L. curasoae is a migratory bat species that pollinates columnar cacti in north-western Mexico, southern Arizona, and south-western New Mexico but may have non-migratory populations in Mexico. We collected 137 samples from six sites: two in north, one in the west, and three in south-central Mexico. The RAPD banding pattern of the bats from each site were used to calculate the proportion of polymorphic loci. The average of polymorphic bands for the south-central population was 65% and for the north-west population was 53%. AMOVA was used to obtain the variance between ( VA = 60.84 % ) and within sites ( VB = 39.16 % ) , meaning that the greater variation is contained among sites and lesser variation inside them. The correlation between geographic and genetic distances was analyzed with a Mantel non-parametric test ( r = 0.72 ) , suggesting a structured population for this species. Our results indicate the presence of two well differentiated populations of L. curasoae, one in south-central Mexico and the other along the Pacific coast ranging from northern Mexico, including Baja California, Sonora, and Jalisco.

Cryptic Species in the <I>Anopheles</I> (<I>Nyssorhynchus</I>) <I>albitarsis</I> (Diptera: Culicidae) Complex: Incongruence Between Random Amplified Polymorphic DNA-Polymerase Chain Reaction Identification and Analysis of Mitochondrial DNA <I>COI</I> Gene Sequences

Random amplified polymorphic DNA (RAPD) diagnostic bands are one tool used to differentiate cryptic mosquito species in the Anopheles albitarsis Complex. Monophyly of four species (A. albitarsis Lynch-Arribálzaga, A. albitarsis B, A. deaneorum Rosa-Freitas, and A. marajoara Galvão & Damasceno) currently identified with the RAPD technique was assessed using sequences of the cytochrome oxidase I (COI) mitochondrial DNA (mtDNA) gene. Maximum parsimony, maximum likelihood, and Bayesian analyses support monophyly for A. albitarsis s.s., A. albitarsis B, and A. deaneorum. Anopheles marajoara, as identified by RAPD banding patterns, was either polyphyletic or paraphyletic in all phylogenetic analyses. The phylogenetic pattern and within-species genetic distances observed in A. marajoara suggest the existence of a previously unidentified species (species E) in northern Brazil and Venezuela. Diagnostic RAPD bands were unable to distinguish between A. marajoara and species E, probably because of the low number of correlated bands used to identify species and weaknesses of the RAPD technique, in particular, violations of the untested assumption of homology of comigrating bands. A. marajoara (even without species E) is paraphyletic with respect to A. deaneorum; if A. deaneorum is a separate species from A. marajoara, then A. marajoara may consist of two or more species in Amazonian Brazil. Based on mtDNA COI sequences, there are at least four phylogenetic species within the Albitarsis Complex: A. albitarsis s.s., A. albitarsis B, A. marajoara, and species E; the species status of A. deaneorum is ambiguous.

Growth strategy of the stoloniferous rattan Calamus javensis in Mt. Halimun, Java

Calamus javensis is a widespread understorey rattan in Southeast Asia and is one of the dominant rattan species in Mt. Halimun National Park, West Java. This species can establish clumps either sexually with seeds or clonality with stolons. To analyze the role of sexual reproduction and clonality for clump establishment in C. javensis, we studied the vegetative and genetic structures of the population. Totals of 2,777 ramets comprising 1,321 clumps were found in three 0.16 ha plots. The number of ramets per clump showed reverse J-shaped distribution with a maximum of 15. Clustering and stoloniferous ramets accounted for 62 and 20% of all ramets, respectively. This indicates that C. javensis is actively engaged in clustering clonal growth. The longest stem was 14 m although 75% of all stems were shorter than 1 m long. The stolon ranged from 6 to 538 cm in length, which decreased the local density of ramets. Using randomly amplified polymorphic DNA (RAPD) analysis, we obtained 56 polymorphic bands for 118 clumps sampled from one of the plots. All samples showed different RAPD banding patterns, except one pair at a distance of 225 cm from each other. The genetic similarity among samples closer than 10 m was significantly higher than the expected value from a purely random distribution. These results suggest that most separate clumps are established by recruitment from seeds, and most seeds dispersed near mother plants. Hence, in C. javensis, clonality is more a growth strategy to increase the size of the genet than a dispersion-propagation strategy to expand the habitat.