Characterization of Vitis germplasm using random amplified polymorphic DNA markers

Summary An analysis of the amplification fragments polymorphism of DNA coming from different accessions of germplasm belonging to species and cultivars of the genus Vitis, was carried out using 40 primer decamers of arbitrary sequence. The RAPD profiles showed a great intraspecific diversity. In many cases a single primer produced a unique pattern for each species. A phylogram tree based upon presence/absence data of the principal DNA bands divided the species according to their geographical origins. The intraspecific polymorphism of DNA fragments was not sufficient for an unambiguous identification of Vitis vinifera cultivars but the RAPD profiles turned out to be highly reproducible. The high capacity of this technique to generate DNA markers offers a new possibility for the study of the genetic relationships in the genus Vitis.Abbreviations PCR Polymerase chain reaction - RAPD Random amplified polymorphic DNA     

Genetic variation of Ethiopian and Eritrean sorghum (Sorghum bicolor (L.) Moench) germplasm assessed by random amplified polymorphic DNA (RAPD)

The extent and patterns of distribution of genetic variation among 80 sorghum (Sorghum bicolor (L.) Moench) germplasm accessions from Ethiopia and Eritrea were investigated using RAPD with 20 oligonucleotide primers. The primers generated a total of 147 polymorphic bands across the 80 accessions with a mean of 7.35 bands per primer. Estimation of the extent of variation by the Shannon-Weaver diversity index revealed an intermediate level of overall variation (H = 53), although the levels varied among regions of origin of the accessions. Partitioning of the total variation revealed considerable variation (77%) within the region of origin of the accessions and the remainder (23%) among regions of origin. Similarly, a large portion (94%) of the total variation was found within the adaptation zones compared to among the adaptation zones (6%). The results suggest a weak differentiation of the sorghum material both on regional and agro-ecological bases, which could be ascribed to the high rate of outcrossing in cultivated sorghum and its free natural hybridization with its wild and weedy relatives, as well as to seed movement by humans. The average genetic dissimilarity was found to be 36% among the 80 accessions and 13% among the 15 regions of origin. Cluster analysis failed to group accessions of the same region or the same adaptation zone, which further confirmed the weak differentiation of the material studied. The clustering pattern of the regions of origin was broadly concordant with previous clustering patterns obtained using morphological characters, in which regions with broad agro-climatic conditions were grouped together.     

Genetic structure among earthworms (Lumbricus terrestris L.) from different sampling sites in western Germany based on random amplified polymorphic DNA

Earthworms are being used as bio-indicators to assess terrestrial pollution. However, it is often not known whether their populations possess a uniform genetic structure, which would allow comparison of residues or biological properties of earthworms from different sampling locations. In order to investigate this point, random amplified polymorphic DNA (RAPD) variation was surveyed in earthworms (Lumbricus terrestris) from five different sampling sites in Germany. Forty oligonucleotide RAPD primers (10 base pairs in length) were screened, three of which produced high polymorphic band patterns. A total of 61 DNA fragments were detected in 90 individuals of L. terrestris from five sampling sites with 49 (80.3%) RAPD markers being polymorphic. The genetic similarities within (band sharing rates between 0.756 and 0.795) and among the L. terrestris populations (0.635) were similar even at widely separate locations. Inter-population variation in the RAPD pattern for all five earthworm populations accounted for 37.9% of the total variation, while intra-population variation for three adjacent Saarland populations accounted for only 18.0% of the total variation. Principal component analysis (PCA) and the genetic distances of the populations confirm these results. Twenty-four percent of the genetic distance is caused by geographical isolation as shown by a test for isolation by distance. These results show that L. terrestris fulfils the genetic qualifications for a bio-indicator particularly at closely located sampling sites. However, the results also suggest that earthworm studies of widely separated locations should include genetic characterisation of the earthworm samples.     

Genetic variation in cucumber (Cucumis sativus L.) as assessed by random amplified polymorphic DNA1

Isozyme and restriction fragment length polymorphisms (RFLPs) have been applied to studies of genetic relationships and germplasm management in cucumber (Cucumis sativus L.). However, isozymes identify relatively few polymorphisms, and RFLPs are technically complex, expensive, and not compatible for the high through-put required for rigorous assessment of this narrow-based germplasm. Since random amplified polymorphic DNA (RAPD) markers do not manifest such shortcomings, a study was conducted in cucumber to examine genetic relationships in diverse germplasm, assess the usefulness of RAPD markers in distinguishing elite accessions, and compare the relative effectiveness of RAPD markers to that of isozyme and RFLP markers. One hundred and eighteen C. sativus accessions were analyzed using variation at 71 RAPD loci (44 mapped and 27 unmapped). Genetic distances among accessions were estimated using the simple matching coefficient complement, and analyzed using multi-dimensional scaling. Each accession had a unique marker profile, indicating that RAPD analysis was useful in genotypic differentiation. Germplasm grouping patterns were consistent with individual accession origins, theoretical dispersal routes and discriminating morphological characters (i.e., sex expression and fruit length to diameter ratio). Although elite accessions were discriminated by RAPD profiling, their genetic distances were relatively small (between 0.01 and 0.58), indicating limited genetic diversity in this germplasm array. Assessment of a subset of the germplasm array using RAPDs resulted in genetic distance measurements more similar to published genetic distance estimates by RFLP markers (Spearman rank correlation, r_s = 0.7–0.8) than estimates by isozyme markers (r_s = 0.4). Data indicate that RAPD markers have utility for analysis of genetic diversity and germplasm management in cucumber.     

Genetic diversity of <Emphasis Type="Italic">Guizotia abyssinica</Emphasis> (L. f.) Cass. (Asteraceae) from Ethiopia as revealed by random amplified polymorphic DNA (RAPD)

Genetic diversity of 70 populations of niger (Guizotia abyssinica) representing all its growing regions in Ethiopia was investigated using random amplified polymorphic DNA (RAPD) to reveal the extent of its populations genetic diversity. Ninety-seven percent of the loci studied was revealed to be polymorphic for the whole data set. The within population diversity estimated by Shannon diversity index and Nei gene diversity estimates was revealed to be 0.395 and 0.158, respectively. The extent of genetic variation of populations from major niger producing regions was significantly lower than that of populations from other regions; however, it is distributed regardless of altitude of growth. Genetic differentiation between populations was estimated with Shannon index as G^′ _ST (0.432), Nei’s G _ST (0.242) and AMOVA based F _ST (0.350) and appears to be equivalent to the average values calculated from various RAPD based studies on outcrossing species. Higher proportion of the variation detected by AMOVA resided within populations (64.58%) relative to the amount of variation among populations (35.42%). UPGMA cluster analysis showed that most of the populations were clustered according to their region of origin. However, some populations were genetically distant from the majority and seem to have unique genetic properties. It is concluded that the crop has a wide genetic basis that may be used for the improvement of the species through conventional breeding and/or marker assisted selection. Collection of germplasm from areas not yet covered and/or underrepresented is the opportunity to broaden the genetic basis of genebank collection.     

Genetic diversity in melon (Cucumis melo L.): Anevaluation of African germplasm

The genetic diversity among 126 exotic (108) andreference array (RA) melon (Cucumismelo L.) accessions (18) was assessed byvariation at 49 random amplified polymorphic DNA marker bands(putative loci) using 29 10-mer primers. Africanaccessions of unknown melon market classes were compared to the RAaccessions from a broad range of C.melo subsp. melo groups(Cantalupensis, Conomon, Inodorus and Flexuosus). Althoughdifferences in groupings occurred after multidimensional scaling andcluster analysis, both analyses placed African accessions into twogroups, which were separate from RA groupings. One African group of33 accessions containing accessions from Zimbabwe (5),Zambia (24), Mali (1), one of two Senegalaccessions and two of three South African accessions examined. Thesecond group, which consisted of 67 accessions containing collectionsfrom Egypt (40), Tunisia (6), Libya(13), Morocco (1), Algeria (2),Ethiopia (1), Niger (1), Sierra Leone(1), S. Africa (1), Zambia (1) andZimbabwe (1). Depending on the multivariate analysistechnique employed, accessions from Kenya, Senegal and Ghana formedeither unique groupings or were grouped with accessions(Cantalupensis) from the RA. Both analyses indicate thatthe genetic differences inherent between the African gene pools isassociated with the geographic proximity of African countries(northern vs. central-southern Africa) in thegermplasm array examined. Moreover, these data indicate that thegenetic diversity of U.S. and European commercial RA germplasm(Cantalupensis and Inodorus) could be enhanced by theintroduction of genetic variation from African accessions, and thatit would be advantageous to acquire more accessions from thisgeographically and ecologically varied region to ensure the retentionof existing genetic diversity.     

Analysis of genetic diversity in Indian taro [Colocasia esculenta (L.) Schott] using random amplified polymorphic DNA (RAPD) markers

Taro [Colocasia esculenta (L.) Schott] germplasm accessions collected from different parts of India were subjected to RAPD (Random Amplified Polymorphic DNA) analysis to assess the genetic diversity prevalent and also to test the genetic basis of morphotypic classification. Thirteen random decamer primers out of the 22 tested were used to analyse 32 taro accessions belonging to 28 morphotypes. Three out of these thirteen primers analysed showed 100 per cent polymorphism. Per cent polymorphism varied from 60 to 100 among the polymorphic primers. High genetic diversity was revealed as the similarity coefficient values ranged from 0.50 to 0.98. No two accessions analysed in the present study showed a similarity coefficient value of one thereby indicating their distinctness and presence of high genetic diversity in Indian taro germplasm. Dendrogram obtained from UPGMA analysis grouped 32 accessions in four clusters and three accessions were placed as outliers. Clustering pattern did not show any strict relationship with geographical distribution, morphotype classification and genotypic diversity. Further, accessions classified, as belonging to the same morphotypic group did not always cluster together. Presence of a very close genepool of the wild, weedy and cultivated forms with extreme levels of phenotypic and genotypic variation is suggested as the reason for high genetic diversity reported. Usefulness of DNA markers such as RAPD in characterising and assessing the genetic diversity in Indian taro germplasm is hereby demonstrated.     

Genetic diversity of Pakistan wheat germplasm as revealed by RAPD markers

Wheat breeding in Pakistan started in 1930s before partition in the United India and so far has released more than 68 cultivars, but no systematic analyses of the genetic diversity of Pakistan wheat have been made. Twenty Pakistan wheat cultivars released from 1933 to 2002 were examined for genetic diversity and relationships using random amplified polymorphic DNA (RAPD) markers. Forty-two RAPD primers were applied and 184 polymorphic bands were generated for each cultivar. Most of the cultivars were genetically interrelated, although six of them displayed some genetic distinctness. The RAPD variation observed among these cultivars was low. Only 40.7% of the total scorable bands were polymorphic, and 26.1% of the polymorphic bands were observed most frequently (f = 0.95) among the 20 cultivars. The proportions of polymorphic bands for each cultivar ranged from 0.67 in ‘Yecora’ to 0.84 in ‘C-250’ with an average of 0.76. About 1.4% of the RAPD variation might have been fixed over the 69 years of wheat breeding, but such fixation was not statistically significant. These results are significant for future improvement and conservation of Pakistan wheat.     

Genetic diversity in African cucumber (Cucumis sativus L.) provides potential for germplasm enhancement

Genetic diversity among 26 cucumber (Cucumis sativus L. var. sativus) accessions from five African countries [Algeria (1), Egypt (21), Ethiopia (2), Kenya (1), and Libya (1)] present in the U.S. National Plant Germplasm System (NPGS) were examined by assessing variation at 71 polymorphic random amplified polymorphic DNA (RAPD) loci. Genetic distances (GD; simple matching coefficient) were estimated among these African accessions and a reference array (RA) of 21 accessions representative of the genetic variation in cucumber. GD among African accessions ranged between 0.41 and 0.97. GD among accessions in the reference array ranged between 0.36 and 0.88. Multivariate analysis identified three distinct groupings (1–3) of African accessions; Group 1 contained 21 accessions (Egypt, Ethiopia and Libya), Group 2 consisted of two accessions (Kenya, Algeria), and Group 3 possessed three accessions (Egypt). These groupings were distinct from each other (P > 0.001). Accessions in Group 1 differed genetically from all other accessions examined (P > 0.01), and accessions in Groups 2 and 3 were uniquely associated with several RA accessions. While GD among accessions in Group 1 ranged between 0.52 and 0.90, distances among Group 2 accessions varied between 0.93 and 0.97. The GD between the two accessions in Group 3 was 0.65. An accession from Syria (PI 181874) and from one Turkey (PI 199383) were genetically more similar to accessions in Group 1 than to other accessions in the RA. Likewise, accessions in Group 2 were genetically similar to two RA accessions from China and a European glasshouse cucumber line, and Group 3 accessions showed genetic affinities with the U.S. market class cultivar Dasher II. Data suggest that some Egyptian accessions (Group 1) possess unique genetic variation, that this germplasm has potential for broadening the genetic base of commerical cucumber, and that further collection of African germplasm is likely to enhance genetic diversity of cucumber in NPGS.     

Genetic diversity in cowpea [Vigna unguiculata (L.) Walp.] as revealed by RAPD markers

The present study, using RAPD analysis, was undertaken to characterize genetic variation in domesticated cowpea and its wild progenitor, as well as their relationships. The materials used consisted of 26 domesticated accessions, including accessions from each of the five cultivar-group, and 30 wild/weedy accessions, including accessions from West, East and southern Africa. A total of 28 primers generated 202 RAPD bands. One hundred and eight bands were polymorphic among the domesticated compared to 181 among wild/weedy cowpea accessions. Wild accessions were more diverse in East Africa, which is the likely area of origin of V. unguiculata var. spontanea. Var. spontanea is supposed to have spread westward and southward, with a loss of variability, loss counterbalanceed in southern Africa by introgressions with local perennial subspecies. Although the variabilty of domesticated cowpea was the highest ever recorded, cultivar-groups were poorly resolved, and several results obtained with isozyme data were not confirmed here. However primitive cultivars were more diverse than evolved cultivars, which still suggests two consecutive bottlenecks within domesticated cowpea evolution. As isozymes and AFLP markers, although with a larger number of markers, RAPD data confirmed the single domestication hypothesis, the gap between wild and domesticated cowpea, and the widespread introgression phenomena between wild and domesticated cowpea.     

腰果树体养分分布特征及其再吸收效率

以5个8龄腰果品系为对象,研究了腰果树体养分分布特征、品系间差异以及元素间相关性;并以无性系FL30为对象,研究了腰果树养分再吸收规律。试验结果表明,N、K、Ca、Mg、Mn主要存在于腰果叶片中,P、Cu主要分布于枝条和根系中,Fe、B则主要分布于根系中,Zn在叶、枝、根中分布较均匀。不同品系腰果树体养分分布特征存在一定的差异。腰果树体中养分元素之间存在一定的作用和影响,叶片中的Ca、Mn、B彼此之间以及N-P、Mg-Mn之间呈极显著的正相关关系,N-K、P-K、Cu-K、Zn-Cu之间呈显著的正相关关系;枝条中的N、P、K彼此之间以及Ca-B、Mg-B、Ca-Mn、Fe-Zn之间呈显著的正相关关系;根系中N-K、Ca-K、B-K、Fe-Zn之间呈极显著的正相关关系,N-B、Ca-B以及K-Zn之间呈显著的正相关关系。腰果树N、P、K再吸收效率较高,Cu、Zn再吸收效率较低;Ca、Mg、Fe、Mn、B在衰老叶片中富集,其中Mn的富集程度最高,其次是Ca>B>Mg,Fe的富集程度最低。     

Genetic variation in captive population of chinese alligator, Alligator sinensis, revealed by random amplified polymorphic DNA (RAPD)

Chinese alligator (Alligator sinensis) is an endemic species in China. The likely extinction of it in the wild has been recognised. To prevent this species becoming extinct, the Anhui Research Centre of Chinese Alligator Reproduction (ARCCAR) was established in Xuanzhou, Anhui Province in 1979, where has been established the largest captive population of Chinese alligator (XZSP) in the world. Another farm (CXSP) was established by villagers in Changxin, Zhejiang Province. The results of an investigation of the two captive subpopulation structures by genetic analysis are presented in this paper. We examined the genetic variation in the two captive subpopulations using RAPDs. Thirty-one random primers were selected among 199 random primers screened. A total of 193 reproducible RAPD fragments were scored among 43 individuals, of which 21 (10.88%) were polymorphic. The genetic distances between 43 individuals ranged from 0 to 0.0376 with average of 0.0104±0.0055 S.E. The genetic similarity in CXSP (0.9948±0.0029 S.E.) was higher than that in XZSP (0.9894±0.0055 S.E.). The founder effect is a possible explanation for very low genetic variation in CXSP. Analysis of the RAPD data showed that the mean phenotypic band frequencies of each polymorphic loci was 0.6656±0.3730 S.E. The lowest phenotypic band frequency (0.0233) was found in four of those polymorphic loci. There was no genetic difference between the two subpopulations (D_ij=0.0009). According to the dendrogram and the distribution of polymorphic fragments in two subpopulations, CXSP originated genetically from XZSP. This paper summarises a preliminary research on genetic structure in populations of Chinese alligator. Although there is higher genetic similary (0.9896±0.0055 S.E.) in captive population of A. sinensis, we did not determine whether or not loss of genetic variation had occurred in relation to a wild control population. The data of malformed offspring will be collected carefully, and wild samples be added to set up a control population in future study.     

Genetic relationships and diversity among Brazilian cultivars and landraces of common beans (Phaseolus vulgaris L.) revealed by AFLP markers

The genetic variation and relationships among 31 accessions of Phaseolus vulgaris L., and two representatives of Vigna unguiculata L., were evaluated by AFLP analysis. A total of 263 DNA fragments across all materials were scored using nine primer combinations, averaging 32 per primer. More than 95% of the amplification products showed polymorphism, indicating high variation at the DNA level among these accessions. Pair-wise genetic similarity (Jaccard's coefficient) ranged from 0.553 to 0.840, with a mean of 0.765. Twenty-three accessions (70%) clustered into three groups. A majority of the commercial cultivars (91%) clustered within a single group, whereas the landraces were distributed along all the variation. An apparent correlation with phaseolin types was detected. Results of this study suggest that Brazilian landraces truly represent the overall genetic variability of Phaseolus vulgaris, confirming the multiple origins of these materials, and their potential as a source of variation for breeding programs.     

Genetic diversity of fast-and slow-growing soybean rhizobia determined by random amplified polymorphic DNA analysis

The genetic relationships among six strains of rhizobia, including three strains of Rhizobium fredii and three strains of Bradyrhizobium japonicum, was determined using random amplified polymorphic DNA (RAPD) technique. In this study, 46 arbitrary 10mer primers were employed for RAPD, generating a total of 251 informative fragments. A dendrogram of phylogenetic relationships among the six strains was constructed. The results indicated that geographical distribution may affect phylogeny, as there were closer relationships among the four Taiwanese strains, SB138, SB562, SB368 and SB651, than between these strains and USDA192, which originated from mainland China. The strain USDA110, obtained from the United States, was used in the parsimony analysis. The greatest similarity (55.6%), existed between two strains of B. japonicum, SB562 and SB138, which both, and the lowest R. fredii (44.4%) between two strains of R. fredii, SB368 and USDA192. We also found a RAPD marker specific to the four Taiwanese SB strains used in the study. The RAPD technique is a potential tool for the identification of the genetics and systematics of different populations. Received: 23 January 1997     

Characterization of genetic variation and relationships among Choix germplasm accessions using RAPD markers

Choix, a plant in the tribe Maydeae of the grass family, has been cultivated in Asia for several thousand years. It is a potential gene resource for improvement of other cereal crops because of its nutritional value and tolerance to stress. Genetic variation and relationships among 21 Choix lachryma-jobi L. accessions were characterized by random amplified polymorphic DNA (RAPD) markers. A total of 205 DNA fragments across all materials were amplified with 31 random primers, averaging 6.61 per primer. Among amplified fragments, 115 showed polymorphism averaging 3.71 per primer. Of amplified markers, 56.1% were polymorphic, indicating considerable variation at the DNA level among these accessions. Some fragments were accession-specific. Pair-wise genetic similarity (GS) among 21 accessions ranged from 0.809 to 0.301. The 21 accessions clustered into two major groups. Three exotic Choix accessions clustered together. Three other Choix accessions, collected from Guangxi, China, clustered into a cohesive subgroup. Four wild types of Choix clustered into the same subgroup. These results indicated that the classification by RAPD data reflected the differences in geographic origins and evolution in Choix.     

Analysis of genetic diversity in a sweet potato (Ipomoea batatas L.) germplasm collection from Tanzania as revealed by AFLP

Sweet potato (Ipomoea batatas L.) is the fifth most important crop in the developing countries after rice, wheat, maize and cassava. The amplified fragment length polymorphism (AFLP) method was used to study the genetic diversity and relationships of sweet potato accessions in the germplasm collection of Sokoine University of Agriculture, Morogoro and Sugarcane Research Institute, Kibaha, Tanzania. AFLP analysis of 97 sweet potato accessions using ten primer combinations gave a total of 202 clear polymorphic bands. Each one of the 97 sweet potato accessions could be distinguished based on these primer combinations. Estimates of genetic similarities were obtained by the Dice coefficient, and a final dendrogram was constructed with the un-weight pair-group method using arithmetic average. AFLP-based genetic similarity varied from 0.388 to 0.941, with a mean of 0.709. Cluster analysis using genetic similarity divided the accessions into two main groups suggesting that there are genetic relationships among the accessions. Principal Coordinate analysis confirmed the pattern of the cluster analysis. Analysis of molecular variance revealed greater variation within regions (96.19%) than among regions (3.81%). The results from the AFLP analysis revealed a relatively low genetic diversity among the germplasm accessions and the genetic distances between regions were low. A maximally diverse subset of 13 accessions capturing 97% of the molecular markers diversity was identified. We were able to detect duplicates accessions in the germplasm collection using the highly polymorphic markers obtained by AFLP, which were found to be an efficient tool to characterize the genetic diversity and relationships of sweet potato accessions in the germplasm collection in Tanzania.     

Genetic differentiation of the parthenogenetic soil collembolan Isotoma notabilis along a copper gradient based on random amplified polymorphic DNA

Five species of collembolans were collected at 24 sampling areas in a copper polluted field. The most abundant species was Isotoma notabilis found at half the sampling sites, in total 490 individuals. Two hundred and fifty-two specimens of these, representing seven sampling areas were analysed by means of random amplified polymorphic DNA using three decamer primers. Two primers each revealed eight phenotypes and the third showed six phenotypes. Distribution of the phenotypes was not homogenous within the sampling area. This result could not be explained as an outcome of either the copper content in the soil or the grass biomass, but was more likely an effect of colonisation from the areas surrounding the field.     

Genetic diversity among Northwestern Argentinian cultivars of common bean (Phaseolus vulgaris L.) as revealed by RAPD markers

The genetic diversity of 10 commercial cultivars of common beans, developed in Northern Argentina was analyzed based on RAPD markers. Sixteen primers were assayed and among them only 4 showed polymorphisms. A similarity matrix was generated by applying three different association coefficients, Simple Matching, Jaccard and Dice. By the UPGMA method dendrograms were generated and also the principal coordinate analysis was performed. The similarity values found were higher than 40% suggesting that genetic diversity is low. Both cluster analysis and principal coordinates analysis associated commercial cultivars either to the Andean or the Mesoamerican gene pool.     

Genetic and Phenotypic Diversity in Downy-mildew-resistant Sorghum (Sorghum bicolor (L.) Moench) Germplasm

Genetic and phenotypic diversity among randomly selected 36 downy-mildew-resistant sorghum accessions were assessed, the former using 10 simple sequence repeat (SSR) marker loci and the latter using 20 phenotypic traits. The number of alleles (a _j ) at individual loci varied from five to 14 with an average of 8.8 alleles per locus. Nei's gene diversity (H _j ) varied from 0.59 to 0.92 with an average of 0.81 per locus. High gene diversity and allelic richness were observed in races durra caudatum (H _j = 0.76, a _j = 4.3) and guinea caudatum (H _j = 0.76, a _j = 3.8) and in east Africa (H _j = 0.78, a _j = 7.2). The regions were genetically more differentiated than the races as indicated by Wright's F _st. The pattern of SSR-based clustering of accessions was more in accordance with their geographic proximity than with their racial likeness. This clustering pattern matched little with that obtained from phenotypic traits. The inter-accession genetic distance varied from 0.30 to 1.00 with an average of 0.78. Inter-accession phenotypic distance varied from 0.01 to 0.55 with an average of 0.33. Eleven accession-pairs had phenotypic distance of more than 0.50 and genetic distance of more than 0.70. These could be used as potential parents in a sorghum downy mildew resistance-breeding program.     

Ploidic and Molecular Analysis of ‘Morado de Huetor’ Asparagus (Asparagus officinale L.) Population; A Spanish Tetraploid Landrace

A study has been conducted into the genetic variation analysis of the tetraploid asparagus landrace ‘Morado de Huetor’ and its relationship with current commercial cultivars using Random Amplified Polymorphic DNA (RAPD) molecular markers. The presence of different ploidic levels in the ‘Morado de Huetor’ landrace has been also studied using cytogenetic analysis. Ten decamer oligonucleotides were used to obtain RAPD markers and to characterise 52 individuals of ‘Morado de Huetor’, 7 of the tetraploid cultivar ‘Purple Passion’, and 55 of 5 diploid hybrid commercial cultivars. Jaccard similarity index was calculated and a cluster analysis using UPGMA (Unweighted Pair-Group Method using Arithmetic Average) was performed. Tetraploid asparagus, ‘Morado de Huetor’ and ‘Purple Passion’, were well differentiated from the diploid ones showing a higher genetic variability. This result suggests a different origin for tetraploid and diploid asparagus varieties. Within the diploid cultivars a variety distribution was obtained. A specific monomorphic band (OPB20₈₈₃) was found in tetraploid varieties. Within the diploid cultivars, two bands (OPB20₈₃₀, OPC15₇₀₅) can be used to distinguish between the hybrid cultivars used in this study. Tri-, penta-, hexa-, and octoploid plants were found in ‘Morado de Huetor’. The origin of these ploidic levels is discussed and their use in the development of new varieties is proposed. In conclusion, ‘Morado de Huetor’ is a genetic resource that could be used to increase the narrow genetic background reported in diploid asparagus cultivars.