Genetic diversity of wheat wild relatives in the Near East detected by AFLP

In order to reveal the molecular genetic diversity of wheat wild relatives, an AFLP analysis was conducted with 16 accessions of five Triticum andAegilops species originating from the Near East. Variation within population was studied with at least seven individuals per accession. Four primer combinations were used for selective amplification. Based on the scored bands, we estimated percentage of polymorphic bands, 1 – proportion of shared bands (1-psb) and nucleotide diversity (π). Of the five species used in this study, Ae. speltoides had the highest level of `within population' variation. This species had also the highest value of the variation among populations. As for Triticum species, the level of variation within population was low in diploid species (T. urartu and T. boeoticum),whereas two tetraploid species (T. dicoccoides and T. araraticum) had relatively high levels of variation within population. While the two diploid Triticum indicated a clear interspecific divergence, the two tetraploid wild wheats were not clearly divergent in this study. The variance portioning analysis indicated that the variation detected for diploid Triticum species was mainly composed of `between species' variation, on the other hand that for tetraploid Triticum was mostly composed of `within population' variation. In conclusion, AFLP analysis reveals molecular variation in all accessions used in this study, suggesting a potential genetic diversity of the wheat wild relatives in natural populations. These results have implications for the design of strategies to maintain genetic diversity within genebank collections. This revised version was published online in August 2006 with corrections to the Cover Date.     

Species relationships in the subgenus Ceratotropis (genus Vigna) as revealed by RAPD analysis

Summary The genetic variation among 23 accessions of 5 species in the subgenus Ceratotropis, genus Vigna, were investigated by random amplified polymorphic DNA (RAPD) analysis. A total of 404 fragments amplified with 24 primers were scored and analyzed by cluster analysis. The accessions used were separated into two main groups with an average of 70% differences. Within the main groups, five subgroups were recognized, which are in complete agreement with taxonomic species. Wild forms were always grouped with their most closely related cultivated forms and they showed variation in each species. The largest intraspecific variation was found in V. radiata (mungbean), in which wild forms (V. radiata var. sublobata) were highly different from each other and from cultivated forms. V. angularis (adzuki bean) showed the least variation and thus, was probably differentiated in relatively recent times.     

Genetic diversity of wild and cultivated soybeans growing in China revealed by RAPD analysis

To evaluate the genetic diversity and to clarify the genetic relationships of wild and cultivated soybeans growing in China, 21 wild soybean accessions and 27 cultivated soybean landraces were analysed by using the random amplified polymorphic DNA method. The data show that wild soybean has a higher genetic variation than cultivated soybean, indicating that genetic variation has been reduced by domestication of wild soybean. Based on Nei's genetic similarity coefficient, all the accessions were classified into two major clusters, corresponding to wild and cultivated varieties of soybean. Furthermore, within each species, the accessions tend to form sub‐clusters that are in agreement with their geographical origins, demonstrating that an extensive geographical genetic differentiation exists in both species. For cultivated soybean, the varieties from the same geographical region but with different seasonal types were found to have closer genetic relationships than varieties from different geographical regions but with the same seasonal type. This result indicates that geographical differentiation plays a key role in the genetic differentiation of both wild and cultivated soybeans. Cultivated soybean varieties with different seasonal types in a region might have been established mainly from the local genotypes.     

Genomic variation and genetic relationships in Ipomoea spp.

Random amplified polymorphic DNA (RAPD) markers were used to develop genetic fingerprints and analyse genetic relationships among 29 Ipomoea accessions from different geographical locations around the world, including unique wild species, and reproducible profiles were obtained for all accessions using random decamer primers. The primers generated 46 polymorphic markers, one primer alone having 10 products, enabling the discrimination of all 29 accessions. A high level of genetic variability in sweet potato collections was suggested by the degree of polymorphism. Half of the Japanese land races were closely related while accessions from Papua New Guinea and The Philippines were distinct and exhibited the greatest genetic diversity. The wild species Ipomoea gracilis and Ipomoea tiliacea formed a group distinct from the cultivated sweet potato. The wild tetraploid accession K233 and the species Ipomoea trifida were progressively more related genetically to the cultivated sweet potato and are the probable progenitors of Ipomoea batatas, and may be suitable as germplasm for genetic enhancement. RAPDs proved to be useful for sweet potato systematics and should be valuable for germplasm management, gene tagging and efficient choice of parents in breeding programmes.     

Genetic variability in melon based on microsatellite variation

A set of 18 simple‐sequence repeat (SSR or microsatellite) markers was used to study genetic diversity in a collection of 27 melon (Cucumis melo L.) accessions, representing a broad range of wild and cultivated melons. The materials studied were highly polymorphic for SSRs and a total of 114 alleles were detected (average of 6.3 alleles per locus). Cluster analysis suggests the division of these accessions into two major groups, largely corresponding to the division of C. melo in the two subspecies agrestis and melo. The assignment of the accession to the subspecies was generally in agreement with published reports, except for those corresponding to the ‘dudaim’ and ‘chito’ cultivar groups, which, according to the observed SSR variability, should be included in subspecies agrestis. Based on cluster analysis, five groups of accessions were defined. The two most divergent groups include mainly accessions from the Mediterranean which form one group, and accessions from China, Japan, Korea and India forming the other. Both groups shared a low level of intra‐accession variation compared with the other groups, which suggests an erosion of their genetic variability because of drift and/or inbreeding. The remaining accessions, mainly from Central Africa and India, were more variable and may be an important source of genetic variation for melon breeding.     

Variation in response to infection with Erysiphe graminis hordei and Puccinia hordei in some wild barley populations in a centre of diversity

Summary The Near East Fertile Crescent extending from Iran to Israel is the centre of origin of cultivated barley and a region of great genetic diversity in wild barley, Hordeum spontaneum C. Koch (syn. H. vulgare ssp. spontaneum (C. Koch Thell)). Wild barley accessions collected from different parts of this region were evaluated for their reactions to infection with three isolates of Erysiphe graminis hordei and two of Puccinia hordei. One culture of each pathogen was isolated in Israel and the others, either in Japan or the United States. Out of a total of 330 wild barley accessions collected from 14 sites in Iran, Turkey and Syria, only 18.8% were resistant to the Israeli culture, and 14.8% were resistant to a composite of the Japanese and American cultures of E. graminis hordei. Out of 105 accessions collected from six sites in Iran and Turkey, none was found to be resistant to the Israeli culture and 34.3% were resistant to the American culture of P. hordei. Considerable variation was observed both within and among sites for reactions to infection with different cultures of each of the two pathogens. The results of this study were compared with those of an earlier study involving wild barley accessions from Israel to illustrate the relative importance of different subregions in the Near East Fertile Crescent as sources of new genes for resistance to E. graminis hordei and P. hordei. Implications of these studies for in situ conservation of genetic diversity in wild barley are discussed.     

Random amplified polymorphic DNA (RAPD) variation in Fagopyrum tataricum Gaertn. accessions from China and the Himalayan region

The diversity among 52 landraces and cultivars of tartary buckwheat (Fagopyrum tataricum Gaertn.) and one accession of its wild ancestor, F. tataricum ssp. potanini Batalin, from diverse geographic origins was examined using random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) markers. Eighteen primers produced a total of 240 fragments, of which 153 (63.75%) were monomorphic and 87 (36.25%) polymorphic bands. UPGMA-based pairwise Jaccard’s coefficient of similarity was used to deduce the relationships among 53 genetically diverse accessions. The similarity between cultivated tartary buckwheat accessions ranged from 0.61 to 1.00. Four distinct clusters were formed which corresponded well with the geographic distribution of the tartary buckwheat. Nepalese accessions showed maximum diversity followed by Chinese accessions. Tartary buckwheat accessions from the Himalayan region of northwestern India revealed a narrow gene pool. The wild buckwheat accession did not group with any of the three cultivated tartary buckwheat groups, and formed its own single-entry group. Genetic similarity (0.59) of Chinese buckwheat accessions with the wild ancestor reaffirmed that cultivated tartary buckwheat originated in the Yunnan province of northwestern China. Consistent with some earlier reports, our study demonstrated the usefulness of the RAPD technique for the characterization of plant genetic resources and assessment of diversity between species. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetic diversity in cultivated Osteospermum as revealed by random amplified polymorphic DNA analysis

A collection of 28 Osteospermum clones and cultivated varieties of different origin were evaluated by random amplified polymorphic DNA (RAPD) analysis. All the clones were identified by 12 decamers selected from a set of 30. This is the first characterization by molecular markers of the genetic material of Osteospermum. The level of genetic diversity among genotypes was assayed and all the accessions tested were then classified into six groups by UPGMA cluster analysis; the clustering of genotypes using the RAPD data proved to be in accordance with their breeding group origin. RAPD analysis can therefore be a useful tool for evaluating genetic variability in other Osteospermum germplasm collections for breeding purposes and for protecting intellectual property rights of improved varieties.     

Ribosomal DNA spacer-length polymorphisms in three samples of wild and cultivated barleys and their relevance to the origin of cultivated barley

This study was conducted to provide additional data for evaluating two important issues surrounding the origin of cultivated barley: (i) the level of genetic diversity of the two‐rowed wild barley from Tibet, and (ii) the distribution of rDNA allele 104 in wild and cultivated barleys in the Occidental region. A total of 198 accessions consisting of three distinct samples were used: 82 entries of two‐rowed wild barley from Tibet, 57 accessions of two‐rowed wild barley from 8 countries with a broad range of representation of two‐rowed wild barley in the world, and 59 landrace accessions from four countries representing a part of the barley‐growing areas in the Middle East. These were assayed for rDNA spacer‐length variants (slvs). In all, 27 rDNA space length pheno types were detected, from which 10 slvs were identified as alleles at the two rDNA loci. The two‐rowed wild barley samples from Tibet had the lowest level of genetic variation as evaluated by rDNA polymorphism. Together with results of previous studies, the two wild forms (two‐rowed and six‐rowed) from Tibet could not account for the large genetic diversity observed in the cultivated barley of this region, suggesting that Tibet is unlikely a centre of origin for cultivated barley. In samples from the Occidental region, allele 104 of Rm2 was very rare in wild barley, but occurred at the highest frequency in cultivated barley, while the reverse is the case for allele 107, which is consistent with previous results. The implications of such a contrasting distribution of these rDNA alleles between wild and cultivated barleys in the origin and evolution of cultivated barley were discussed.     

Variation in developmental patterns of wild barley (Hordeum spontaneum L.) and cultivated barley (H. vulgare L.)

Summary Six wild barley (Hordeum spontaneum) accessions, from a diverse range of habitats, and two spring-cultivated barleys, were examined for variation in durations of development phases. The durations of the leaf initiation and spikelet initiation phases were longer and spikelet growth phases shorter, in wild than in cultivated barley. Across all wild and cultivated barleys the rate and duration of spikelet initiation were negatively correlated, but neither was related to the number of spikelets per spike. The number of spikelets was positively correlated with the number of leaves and the ratio of the number of spikelets to the number of leaves declined with increasing time to anthesis, indicating that each successive leaf was associated with a diminishing increase in the number of spikelets. The duration of culm elongation and final culm length were shorter in accessions of cultivated barley compared with wild barley. This paper also discusses the feasibility for increasing the number of spikelets per spike through breeding for genetic changes in lengths of pre-anthesis phases of development.Abbreviations ANOVA Analysis of variance - HV Hordeum vulgare - CE Culm elongation - DR Double ridge - HS Hordeum spontaneu - ° Cd Degree days     

Molecular Marker based Genetic Diversity Analysis in Rice (Oryza sativa L.) using RAPD and SSR markers

The availability of an array of molecular marker systems allowed comparing the efficiency of two of these marker systems to estimate the relationships among various taxa. The objective of this study was to assess the genetic diversity among 40 cultivated varieties and five wild relatives of rice, Oryza sativa L. involving simple sequence repeat (SSR) randomly amplified polymorphic DNA (RAPD) markers. The accessions were evaluated for polymorphisms after amplification with 36 decamer primers and 38 SSR primer pairs. A total of 499 RAPD markers were produced among the 40 cultivated varieties and five wild relatives with a polymorphism percentage of 90.0. Out of 38 SSR primer pairs used, only one locus viz., RM115 was monomorphic. The average Polymorphism Information Content (PIC) value was 0.578 and it ranged from a low of zero (RM 115) to a high of 0.890 (RM 202). The Mantel matrix correspondence test was used to compare the similarity matrices and the correlation coefficient was 0. 582. The test indicated that clusters produced based on RAPD and SSR markers were not conserved since matrix correlation value was 0.582 as against the minimum required value of 0.800. The two marker systems contrasted most notably in pair-by-pair comparisons of relationships. SSR analysis resulted in a more definitive separation of clusters of genotypes indicating a higher level of efficiency of SSR markers for the accurate determination of relationships between accessions that are too close to be accurately differentiated by RAPD markers. This revised version was published online in July 2006 with corrections to the Cover Date.     

Microsatellite (SSR) variation in a collection of Malus (apple) species and hybrids

A collection of 142 accessions of 23 Malus species, derived hybrids and cultivar accessions from the USDA-ARS Plant Genetic Resources Unit's core collection, which represents an extensive range of Malus species, was screened with a set of previously described SSR (simple sequence repeat) markers. The markers were used to determine genetic identities, estimate genetic diversity, identify genetic relationships among the accessions, and determine the utility of SSR primers developed from Malus ×domestica for making genetic assessments across the whole Malus genus. All eight primer pairs amplified multiple fragments when used in polymerase chain reactions with DNA from these accessions. High levels of variation were detected with a mean of 26.4 alleles per locus and a mean direct count heterozygosity across all eight loci equal to 0.623. The eight primer pairs used in this study unambiguously differentiated all but five pairs of accessions in this collection of 142 accessions of 23 Malus species, derived hybrids and cultivars. These SSR data were not useful in identifying genetic relationships among this diverse collection of accessions, with the majority of the accessions not clustering in ways concordant with taxonomic information and/or geographic origin. The resulting phenogram resolved only two meaningful clusters, for the taxonomically isolated Section Chloromeles and for M. fusca accessions, reflecting genetic relationships arising from geographic origin. The detection of identical accessions in the collection, which were previously considered to be unique, highlights the critical need to further bolster collections of certain Malus species. This revised version was published online in July 2006 with corrections to the Cover Date.     

Assessment of genetic relatedness of the genera Ananas and Pseudananas confirmed by RAPD markers

Genetic relationships among 18 accessions, including 16 of Ananas and two of Pseudananas, were investigated using RAPD molecular markers. The procedure for DNA extraction was adapted from the method of Dellaporta et al. (1983) where an incubation in proteinase K and a purification step were included. From the total of 148 markers scored,132 (89.2%) were polymorphic. The similarity matrix was used for cluster analysis. The phenogram developed from the RAPD bands showed that for most of the cases, the accessions within a species grouped together. Nevertheless, a moderate infraspecific genetic variation was observed. For example, DNA data grouped all A. comosus accessions with a mean similarity coefficient of 0.85. Comparable results were obtained with all other species investigated. The highest genetic divergence was found withinA. lucidus where the mean similarity coefficient among accessions was0.75. A similar level of genetic polymorphism was observed among species,therefore, a definition about which species were involved in the constitution of A. comosus genotypes was not possible. These results agree with the breeders standpoint suggesting that all Ananas species belong to the primary gene pool of pineapple. This revised version was published online in July 2006 with corrections to the Cover Date.     

Geographical differentiation and diallel analysis of seed dormancy in barley

Seed dormancy is one of the most important parameters affecting the malting process and pre-harvest sprouting in barley (Hordeum vulgare L.). Variation of seed dormancy in 4365 cultivated and 177 wild barley (ssp. spontaneum) accessions derived from different regions of the world was investigated in Okayama University, Kurashiki, Japan. Seed dormancy of each accession was estimated from their germination percentages at 0, 5, 10 and 15 weeks post-harvest after-ripening periods. All of the wild barley accessions showed less than 10% germination at 0 week after-ripening period. Level of seed dormancy in 4365 cultivated barley accessions showed a clear geographical differentiation. Seventy seven percent of Ethiopian accessions showed high germination percentages, while 86% of Japanese, Turkish and North African accessions showed low germination percentages at 0 week after-ripening period. A half diallel cross using eleven barley accessions with different level of dormancy revealed that seed dormancy was predominately controlled by additive gene effects. These results suggest that large genetic diversity for seed dormancy in barley is explained as different levels of additive accumulation of genetic factors. Barley varieties showing appropriate dormancy could be developed by crossing among barley germplasm accessions used in the present study.     

Genotypic variation in polyphenol content of barley grain

The polyphenol content in pearl barley, which is highly correlated to a browning reaction after heat treatment, was investigated using 1,347 cultivated barley varieties (H. vulgare) and two wild accessions (H. vulgare subsp. spontaneum) collected from different areas of the world. The polyphenol content in the cultivated barley shows a wide variation ranging from 0.19 to 0.75 mg/g with a nearly normal frequency distribution. The polyphenol content in the hulless varieties from Japan and Korea was low. On the other hand, the polyphenol content in wild barley was about two times higher than the average value recorded in cultivated barley. Based on HPLC analysis, five lowest-polyphenol content local varieties do not represent proanthocyanidin-free mutants. This revised version was published online in August 2006 with corrections to the Cover Date.     

Intra- and inter-population variations in grain protein percentage in wild tetraploid wheat,Triticum turgidum var. dicoccoides

Summary Grain protein percentage (GPP) was studied in 910 accessions of the wild tetraploid wheat, Triticum turgidum var. dicoccoides, collected from 22 populations representing different ecogeographical conditions in Israel. High values of GPP were found, ranging from 19.7% to 28.0% for population means, and from 14.1% to 35.1% for single accessions. Marginal populations had usually lower GPP and smaller intra-population variation than central ones. Repeated sampling of some central populations for four consecutive years revealed relatively large intra-population fluctuations. A high and significant genetic component of variation was found within and between populations by a nested analysis of variance in two nurseries. However, the regression coefficients of parents vs. offsprings were relatively low, indicating a smaller genetic component of variation which may be accounted for by a significant genotype × environment interaction. No correlation was found between GPP and ecological factors, except for soil type: accessions growing on terra-rossa had higher GPP than those growing on basaltic soil. Accessions with black glumes, or with glabrous auricles, or with large grains exhibited high GPP values. A strategy for collecting accessions with high GPP is presented, and the potential use of high GPP genotypes in breeding programs is discussed.     

Genetic structure and differentiation in hop (Humulus lupulus L.) as inferred from microsatellites

A set of 67 wild and cultivated hop accessions, representative of hop diversity, was genotyped with 29 SSR markers in order to investigate the population structure and genetic diversity among hop genotypes. A total of 314 alleles was detected, with an average of 10.8 alleles per locus and an average PIC content of 0.607. Model-based clustering placed the accessions into five germplasm groups. A distance-based tree showed good agreement with five germplasm groups, and additionally assigned accessions omitted from model-based analysis into two additional germplasm groups. The 67 hop accessions were thus subdivided in seven germplasm groups, with three corresponding to major breeding groups and four to wild hops. This finding is in accordance with two biogeographically separated hop germplasms (European and North American origin) and with the known history of the accessions. North American hop germplasm was partitioned into native and cultivated germplasm groups. European germplasm was divided into two groups of hop cultivars representing distinguishable European germplasms and three new groups of native hops, which were differentiated for the first time by this analysis. Admixture analysis showed shares of various ancestries in hop cultivars, mostly congruent with pedigree data, and the introgression of various ancestries in some native hops. The above results have so far given the most detailed insight to date into the population structure of hop diversity, which is important for its effective use in hop breeding.     

Determination of genetic variation and taxonomy in lentil (Lens Miller) species by chloroplast DNA polymorphism

Summary Chloroplast DNA restriction fragment lenght polymorphisms (RFLP) were used to examine the taxonomic relationships of cultivated and wild lentil (Lens Miller) species and identify the extent of genetic variation in this genus. Twelve accessions representing all Lens subspecies were digested with four hexanucleotide-recognizing restriction endonucleases. These digests randomly surveyed 540 base pairs, or 0.4% of the approximately 125 kilobase lentil chloroplast genome. A high degree of gragment length conservation was seen among members of crossability group I, i.e., L. c. ssp. culinaris, L. c. ssp. orientalis and L. c. ssp. odemensis. Accessions of the two subspecies comprising crossability group II, i.e., L. n. ssp. nigricans and L. n. ssp. ervoides, showed the greatest amount of variation when compared to the cultivated lentil, L. c. ssp. culinaris. Limited variation was observed within subspecies except for L. n. ssp. nigricans, where accessions of the normal cytotype were highly polymorphic to those of the differentiated cytotype. Chloroplast DNA RFLPs reaffirm hypotheses that propose L. c. ssp. orientalis as the progenitor to the cultivated lentil. The implications of this study on taxonomy and genetic resources is also discussed.     

Genetic differences in growth within and between Lycopersicon species

Summary Growth analyses were carried out on 88 accessions of five Lycopersicon species. Experiments were conducted in a climate room at 19/14° C day/night temperature which was irradiated at 20 W/m² for eight hours per day. Large differences in plant weights between wild species and the cultivated tomato were observed from 44 to 84 days after sowing. The increase in plant dry weight could be described by a second order polynomial function. When compared at a standardized plant weight of one gram, the relative growth rates (RGR) of the wild and cultivated accessions ranged from 5.3 to 11.8% and 8.5 to 12.2% per day respectively, limiting the use of wild species as sources for strong growth. When expressed at plant weights of one and three g large differences in decrease of the RGR were observed within L. esculentum. The modern hybrid tomato cultivars were among the fastest growing genotypes, with a relatively slow decrease in RGR.     

Evidence of a partial reproductive barrier between wild and cultivated pearl millets (Pennisetum glaucum)

Summary The occurrence of seed malformation in association with reduced thousand grain weight and germination ability has been observed in crosses between cultivated female plants and wild male plants. A survey of 16 cultivated accessions (P. glaucum subsp. glaucum) and 11 wild accessions (P. glaucum subsp. monodii) ranging over the whole species diversity showed this postzygotic incompatibility was general, but its intensity varied greatly with the cultivated female accession used and very little with the wild male parent origin. About 15% of the 123 cultivated x wild crosses observed gave normal seeds. Seed malformation has never been observed in crosses between cultivated accessions and appeared independent of genetic distances between the parents. The reciprocal crosses between wild female plants and cultivated male plants gave normal-looking seeds with good germination but consistently reduced thousand grain weight. Both seed malformation and seed small size are an expression of a genetic imbalance. These slight reproductive barriers seem to have been built during the domestication process.Abbreviation ICRISAT International Crop Research Institute for the Semi-Arid Tropics