The genetic characterization of Turkish watermelon (<Emphasis Type="Italic">Citrullus lanatus</Emphasis>) accessions using RAPD markers

Genetic diversity of the Turkish watermelon genetic resources was evaluated using different Citrullus species, wild relatives, foreign landraces, open pollinated (OP) and commercial hybrid cultivars by RAPD markers. The germplasm was consisted of 303 accessions collected from various geographical regions. Twenty-two of 35 RAPD primers generated a total of 241 reproducible bands, 146 (60.6%) of which were polymorphic. Based on the RAPD data the genetic similarity coefficients were calculated and the dendrogram was constructed using UPGMA (Unweighted pair-group method with arithmetic average). Cluster analysis of the 303 accessions employing RAPD data resulted in a multi-branched dendrogram indicating that most of the Turkish accessions belonging to var. lanatus of Citrullus lanatus (Thunb.) Matsum et Nakai were grouped together. Accessions of different Citrullus species and Praecitrullus fistulosus (Stocks) Pangalo formed distant clusters from C. lanatus var. lanatus. Among 303 accessions, a subset of 56 accessions was selected representing different groups and a second dendrogram was constructed. The genetic similarity coefficients (GS) within the Turkish accessions were ranged from 0.76 to 1.00 with 0.94 average indicating that they are closely related. Taken together, our results indicated that low genetic variability exist among the watermelon genetic resources collected from Turkey contrary to their remarkable phenotypic diversity.     

Genetic diversity of <Emphasis Type="Italic">Danthonia spicata</Emphasis> (L.) Beauv. based on genomic simple sequence repeat markers

Danthonia spicata (L.) Beauv., commonly known as poverty oatgrass, is a perennial bunch-type grass native to North America. D. spicata is often found in low input turfgrass areas on the East Coast of the United States and has potential for development as a new native low input turfgrass species. Roche 454 sequenced randomly sheared genomic DNA reads of D. spicata were mined for SSR markers using the MIcroSAtellite identification tool. A total of 66,553 singlet sequences (approximately 37.5 Mbp) were examined, and 3454 SSR markers were identified. Trinucleotide motifs with greater than six repeats and possessing unique PCR priming sites within the genome, as determined by Primer-BLAST, were evaluated visually for heterozygosity and mutation consistent with stepwise evolution using CLC Genomics software. Sixty-three candidate markers were selected for testing from the trinucleotide SSR marker sites meeting these in silico criteria. Ten primer pairs that amplified polymorphic loci in preliminary experiments were used to screen 91 individual plants composed of at least 3–5 plants from each of 23 different locations. The primer pairs amplified 54 alleles ranging in size from 71 to 246 bp. Minimum and maximum numbers of alleles per locus were two and 12, respectively, with an average of 5.4. A dendrogram generated by unweighted pair group method with arithmetic mean cluster analysis using the Jaccard’s similarity coefficient was in agreement with the grouping obtained by Structure v2.3. The analyses were dominated by clonal groupings and lack evidence for gene flow with some alleles present in a single plant from a single location. Fourteen multilocus genotype groups were observed providing strong evidence for asexual reproduction in the studied D. spicata populations.     

Mitochondrial genome diversity in <Emphasis Type="Italic">Beta vulgaris</Emphasis> L. ssp. <Emphasis Type="Italic">vulgaris</Emphasis> (Leaf and Garden Beet Groups) and its implications concerning the dissemination of the crop

Four mitochondrial minisatellites were used to study cytoplasmic diversity in leaf and garden beet germplasm resources. Eleven multi-locus haplotypes were identified, of which one (named mitochondrial minisatellite haplotype 4, hereafter min04) was associated with male-sterile Owen cytoplasm and two others (min09 and min18), with a normal fertile cytoplasm. European leaf beet germplasm exhibited the greatest haplotype diversity, with min09 and min18 predominating. In North African leaf beet accessions, only these two haplotypes were observed, making it likely that North African accessions were descended from European genotypes. The prevalence of min18 was also noted in leaf beet from the Middle East and western Asia. Such a pattern contrasts with that found in east Asian leaf beet where the two haplotypes were extremely rare. The geographical structure of the mitochondrial haplotypes allowed us to infer possible dissemination pathways of leaf beet. Additionally, we showed that mitochondrial genome diversity was low in garden beet germplasm, with min18 being highly predominant. An explanation of this limited diversity may lie in the geographically restricted origin of as well as relatively short cultivation histories of garden beet.     

Genetic relationships of boxwood (<Emphasis Type="Italic">Buxus</Emphasis> L.) accessions based on genic simple sequence repeat markers

Boxwoods (Buxus L., Buxaceae) are popular woody landscape shrubs grown for their diverse forms and broad-leaved evergreen foliage. We used genic simple sequence repeat (genic-SSR) markers to assess genetic diversity and relatedness of 275 accessions from the National Boxwood Collection at the U.S. National Arboretum. Flow cytometry was conducted to determine the relative ploidy of each accession. Genic-SSR loci were highly variable among the accessions, detecting an average of 6.7 alleles per locus based on 17 primer pairs. Data were analyzed with a distance matrix based on Jaccard’s similarity index, followed by Unweighted Pair Group Method with Arithmetic Mean clustering. Two major clusters were identified, with four subclusters consisting of individual accessions from B. balearica Lam., B. bodinieri Lévl., B. harlandii Hance, B. microphylla Siebold et Zuccarini, B. sempervirens L., B. sinica (Rehd. et Wils.) M. Cheng, and their putative interspecific hybrids. The accessions generally clustered by cultivar, provenance, or species. Clustering within each group typically reflected breeding pedigrees, when known, and the clusters were supported by bootstrap results. This information will be used for breeding programs and collection management, and for identifying possible sources of disease tolerance for boxwood blight and other diseases and pests.     

Genetic diversity of the D-genome in <Emphasis Type="Italic">T. aestivum</Emphasis> and <Emphasis Type="Italic">Aegilops</Emphasis> species using SSR markers

Simple sequence repeats (SSRs), highly dispersed nucleotide sequences in genomes, were used for germplasm analysis and estimation of the genetic relationship of the D-genome among 52 accessions of T. aestivum (AABBDD), Ae. tauschii (D^tD^t), Ae. cylindrica (CCD^cD^c) and Ae. crassa (MMD^cr1D^cr1), collected from 13 different sites in Iran. A set of 21 microsatellite primers, from various locations on the seven D-genome chromosomes, revealed a high level of polymorphism. A total of 273 alleles were detected across all four species and the number of alleles per each microsatellite marker varied from 3 to 27. The highest genetic diversity occurred in Ae. tauschii followed by Ae. crassa, and the genetic distance was the smallest between Ae. tauschii and Ae. cylindrica. Data obtained in this study supports the view that genetic variability in the D-genome of hexaploid wheat is less than in Ae. tauschii. The highest number of unique alleles was observed within Ae. crassa accessions, indicating this species as a great potential source of novel genes for bread wheat improvement. Knowledge of genetic diversity in Aegilops species provides different levels of information which is important in the management of germplasm resources.     

<Emphasis Type="Italic">Rhizobium</Emphasis>,<Emphasis Type="Italic"> Bradyrhizobium</Emphasis> and<Emphasis Type="Italic"> Agrobacterium</Emphasis> strains isolated from cultivated legumes

The present study was conducted to isolate and characterize rhizobial strains from root nodules of cultivated legumes, i.e. chickpea, mungbean, pea and siratro. Preliminary characterization of these isolates was done on the basis of plant infectivity test, acetylene reduction assay, C-source utilization, phosphate solubilization, phytohormones and polysaccharide production. The plant infectivity test and acetylene reduction assay showed effective root nodule formation by all the isolates on their respective hosts, except for chickpea isolate Ca-18 that failed to infect its original host. All strains showed homology to a typical Rhizobium strain on the basis of growth pattern, C-source utilization and polysaccharide production. The strain Ca-18 was characterized by its phosphate solubilization and indole acetic acid (IAA) production. The genetic relationship of the six rhizobial strains was carried out by random amplified polymorphic DNA (RAPD) including a reference strain of Bradyrhizobium japonicum TAL-102. Analysis conducted with 60 primers discriminated between the strains of Rhizobium and Bradyrhizobium in two different clusters. One of the primers, OPB-5, yielded a unique RAPD pattern for the six strains and well discriminated the non-nodulating chickpea isolate Ca-18 from all the other nodulating rhizobial strains. Isolate Ca-18 showed the least homology of 15% and 18% with Rhizobium and Bradyrhizobium, respectively, and was probably not a (Brady)rhizobium strain. Partial 16S rRNA gene sequence analysis for MN-S, TAL-102 and Ca-18 strains showed 97% homology between MN-S and TAL-102 strains, supporting the view that they were strains of B. japonicum species. The non-infective isolate Ca-18 was 67% different from the other two strains and probably was an Agrobacterium strain.     

Putative Role of <Emphasis Type="Italic">Flavobacterium</Emphasis>, <Emphasis Type="Italic">Dokdonella</Emphasis> and <Emphasis Type="Italic">Methylophilus</Emphasis> Strains in Paracetamol Biodegradation

Paracetamol, the most widely and globally used analgesic and antipyretic, is easily accumulated in aquatic environments. In the present study, the biodegradation of paracetamol in different media (one for general growth, one specific for sulfate reducing bacteria, a mineral salts medium and municipal wastewater) inoculated with two types of sludge (from anaerobic lagoon and from oxidation ditch) under different oxygenic conditions (anoxic; moderate oxygenation in open flasks and high oxygenation by aeration) was investigated. In addition, bacteria with relative abundances increasing simultaneously with paracetamol degradation, when this drug was the only carbon source, thus with a putative role in its degradation, were identified using 16S rRNA gene sequences. The results show that aerobic microorganisms had a major role in the degradation of paracetamol, with 50 mg/L totally removed from municipal wastewater after 2 days incubation with aeration, and that the metabolites 4-aminophenol and hydroquinone plus one compound not identified in this work were produced in the process. The identification of bacteria with a role in the degradation of paracetamol revealed a strain from genus Pseudomonas with the highest final relative abundance of 21.2%, confirming previous works reporting strains of this genus as paracetamol decomposers. Besides, genera Flavobacterium, Dokdonella and Methylophilus were also in evidence, with initial relative abundances of 1.66%, 1.48 and 0.00% (not detected) in the inoculum and 6.91%, 3.80 and 3.83% after incubation, respectively. Therefore, a putative role of these genera in paracetamol biodegradation is suggested for the first time.

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Graphical Abstract ?

     

Comparison of the genetic diversity between <Emphasis Type="Italic">Triticum aestivum</Emphasis> ssp. <Emphasis Type="Italic">tibetanum</Emphasis> Shao and Tibetan wheat landraces (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) by using intron-splice junction primers

In order to evaluate and compare the germplasm resources of wheat in Tibet, we analyzed the genetic diversity of 136 Triticum aestivum ssp. tibetanum Shao and 119 Tibetan wheat landraces (Triticum aestivum L.) by using Intron-Splice Junction (ISJ) primers. The results showed that polymorphism of PCR products were obtained by 33 primer combinations, which accounted for 11% of the 300 primer combinations produced by 26 ISJ primers. A total of 333 stable bands can be amplified from the T. aestivum ssp. tibetanum Shao and 243 bands were polymorphic, which accounted for 72.9% of the total bands. Tibetan wheat Landraces produced 316 stable bands, of which 197 bands were polymorphic. The polymorphic bands accounted for 62.34% of the total bands produced from Tibetan wheat landraces. The genetic diversity of T. aestivum ssp. tibetanum Shao was higher than that of Tibetan wheat landraces in Tibet, suggesting that T. aestivum ssp. tibetanum Shao can be used as important genetic resource for the breeding and genetic improvement of wheat in Tibet. Matrix (1, 0) was generated according to the presence or absence of the bands produced from a particular wheat accession. Clustering and principle coordinates analysis showed that T. aestivum ssp. tibetanum Shao and Tibetan wheat landraces were divided into two groups. We conclude that high polymorphisms produced by ISJ primers can reflect the genetic diversity between T. aestivum ssp. tibetanum Shao and Tibetan wheat landraces.     

<Emphasis Type="Italic">Achromobacter insolitus</Emphasis> and <Emphasis Type="Italic">Zoogloea ramigera</Emphasis> associated with wheat plants (<Emphasis Type="Italic">Triticum aestivum</Emphasis>)

This study reports for the first time the presence of diazotrophic bacteria belonging to the genera Achromobacter and Zoogloea associated with wheat plants. These bacterial strains were identified by the analysis of 16S rDNA sequences. The bacterium IAC-AT-8 was identified as Azospirillum brasiliense, whereas isolates IAC-HT-11 and IAC-HT-12 were identified as Achromobacter insolitus and Zoogloea ramigera, respectively. A greenhouse experiment involving a non-sterilized soil was carried out with the aim to study the endophytic feature of these strains. After 40 days from inoculation, all the strains were in the inner of roots, but they were not detected in soil. In order to assess the location inside wheat plants, an experiment was conducted under axenic conditions. Fifteen days after inoculation, preparations of inoculated plants were observed by the scanning electron microscope, using the cryofracture technique, and by the transmission electron microscope. It was observed that all isolates were present on the external part of the roots and in the inner part at the elongation region, in cortex cells, but not in the endodermis or in the vascular bundle region. No colonizing bacterial cells were observed in wheat leaves.     

Salt tolerance in <Emphasis Type="Italic">Zea mays</Emphasis> (L). following inoculation with <Emphasis Type="Italic">Rhizobium</Emphasis> and <Emphasis Type="Italic">Pseudomonas</Emphasis>

This study aimed to investigate the effect of inoculation with plant growth-promoting Rhizobium and Pseudomonas species on NaCl-affected maize. Two cultivars of maize (cv. Agaiti 2002 and cv. Av 4001) selected on the basis of their yield potential were grown in pots outdoors under natural conditions during July. Microorganisms were applied at seedling stage and salt stress was induced 21 days after sowing and maintained up to 50% flowering after 120 days of stress. The salt treatment caused a detrimental effect on growth and development of plants. Co-inoculation resulted in some positive adaptative responses of maize plants under salinity. The salt tolerance from inoculation was generally mediated by decreases in electrolyte leakage and in osmotic potential, an increase in osmoregulant (proline) production, maintenance of relative water content of leaves, and selective uptake of K ions. Generally, the microbial strain acted synergistically. However, under unstressed conditions, Rhizobium was more effective than Pseudomonas but under salt stress the favorable effect was observed even if some exceptions were also observed. The maize cv. Agaiti 2002 appeared to be more responsive to inoculation and was relatively less tolerant to salt compared to that of cv. Av 4001.     

Genetic diversity among <Emphasis Type="Italic">Jatropha</Emphasis> species as revealed by RAPD markers

The genus Jatropha is native of tropical America with more than 200 species that are widely distributed in tropics with a promise for use as an oil crop for biodiesel. This investigation was carried out to assess the genetic diversity of 12 Jatropha species based on random amplified polymorphic DNA markers. From 26 random primers used, 18 primers gave reproducible amplification banding patterns of 112 polymorphic bands out of 134 bands scored accounting for 80.2% polymorphism across the genotypes. Three primers viz., OPA 4, OPF 11, and OPD 14 generated 100% polymorphic patterns. The polymorphic information content was highest for the primer OPD 14 (0.50) followed by the primers OPF 11 and OPAD 11 (0.48). Jaccard’s coefficient of similarity varied from 0.00 to 0.85, indicative of high level of genetic variation among the genotypes studied. UPGMA cluster analysis indicated three distinct clusters, one comprising all accessions of J. curcas L., while second included six species viz., J. ramanadensis Ramam., J. gossypiifolia L., J. podagrica Hook., J. tanjorensis J. L. Ellis et Saroja J. villosa Wight and J. integerrima Jacq. J. glandulifera Roxb. remained distinct and formed third cluster indicating its higher genetic distinctness from other species. The overall grouping pattern of clustering corresponds well with principal component analysis confirming patterns of genetic diversity observed among the species. The result provides valid guidelines for collection, conservation and characterization of Jatropha genetic resources.     

DNA markers and pollen morphology reveal that <Emphasis Type="Italic">Praecitrullus fistulosus</Emphasis> is more closely related to <Emphasis Type="Italic">Benincasa hispida</Emphasis> than to <Emphasis Type="Italic">Citrullus</Emphasis> spp.

The round melon Praecitrullus fistulosus (Stocks) Pangalo has been cultivated in Asia since ancient times and has been considered an underexploited crop in the western world. In the USA, there is an increased interest in using P. fistulosus as a commercial vegetable, and possibly as a rootstock for grafting watermelon, melon, or cucumber. However, the taxonomic classification of P. fistulosus is incomplete and for many years it has been considered a close relative of watermelon [Citrullus lanatus subsp. vulgaris (Schrad. ex Eckl. et Zeyh.) Fursa] and was previously classified as Citrullus lanatus subsp. fistulosus (Stocks) Duthie et J.B. Fuller. Here, we used two sets of DNA markers to assess the genetic similarity of P. fistulosus in relation to Citrullus spp. {including Citrullus lanatus subsp. vulgaris, C. lanatus subsp. lanatus, Citroides group [also known as C. lanatus (Thunb.) Matsum. et Nakai subsp. lanatus var. citroides (Bailey) Mansf. ex Greb.], and C. colocynthis (L.) Schrad.}, Cucumis spp. (including C. melo, C. sativus, C. anguria, C. meeusei, C. zeyheri), Benincasa hispida (Thunb.) Cogn., Lagenaria siceraria (Mol.) Standl. and Cucurbita spp. (including C. moschata Duchesne and the winter squash C. maxima Duchesne). The first marker set comprised 501 markers that were produced by 38 primer pairs derived from watermelon expressed sequenced tags (ESTs) containing simple sequence repeat (SSR) motifs (designated as EST-SSR primers; produced 311 markers), and by 18 primer pairs derived from ESTs that do not contain SSR motives (designated here as EST-PCR primers; produced 190 markers). The second marker set comprised 628 markers that were produced by 18 sequence related amplified polymorphism (SRAP) primer pairs. The phylogenetic data indicated that among these cucurbit species, the wax gourd B. hispida is the closest to the P. fistulosus. Pollen observations, using light microscopy, indicated that each of the cucurbit genera examined here has unique pollen morphology. The Cucurbita spp. have globular pollen grains with a stigmatic surface. The L. siceraria has polygonal pollen grains with symmetrical boundaries, while the Citrullus spp. and Cucumis spp. have ovular (conical) and triangular shaped pollen grains (respectively). The B. hispida and P. fistulosus have spherical or semispherical pollen grains. These pollen features appear to be in agreement with the phylogenetic relationships of these two species based on DNA markers. Analysis with 12 SRAP primer pairs revealed low genetic diversity among 18 United States Plant Introductions (PIs) of P. fistulosus, indicating the need to expand the germplasm collection of this cucurbit crop.     

Characterization of fertile amphidiploid between <Emphasis Type="Italic">Raphanus sativus</Emphasis> and <Emphasis Type="Italic">Brassica alboglabra</Emphasis> and the crossability with <Emphasis Type="Italic">Brassica</Emphasis> species

A fertile amphidiploid × Brassicoraphanus (RRCC, 2n = 36) between Raphanus sativus cv. HQ-04 (2n = 18, RR) and Brassica alboglabra Bailey (2n = 18, CC) was synthesized and successive selections for seed fertility were made from F₄ to F₁₀. F₁₀ plants exhibited good fertility with 14.9 seeds per siliqua and 32.3 g seeds per plant. Cytological observation revealed that frequent secondary pairing occurred among 3 chromosome pairs in pollen mother cells of plants (F₄) with lower fertility, but not of plants with high fertility (F₁₀). GISH analysis indicated that these F₁₀ plants included the expected 18 chromosomes from R. sativus and B. alboglabra, respectively, but they lost approximately 27.6% R. sativus and 35.6% B. alboglabra AFLP (amplified fragment length polymorphism) bands. The crossability of the Raphanobrassica with R. sativus and 5 Brassica species (13 cultivars) were investigated. Seeds or F₁ seedlings were easy to be produced from crosses × Brassicoraphanus × R. sativus, and B. napus, B. juncea and B. carinata × Brassicoraphanus. Fewer seeds or seedlings were obtained from crosses × Brassicoraphanus × B. napus, B. juncea and B. carinata. However, few seeds were harvested in the reciprocals of × Brassicoraphanus with B. rapa and B. oleracea. The possible cause of fertility improvements and the potential of the present × Brassicoraphanus for breeding were discussed.     

Cultivation of lotus (<Emphasis Type="Italic">Nelumbo nucifera</Emphasis> Gaertn. ssp. <Emphasis Type="Italic">nucifera</Emphasis>) and its utilization in China

Lotus (Nelumbo nucifera Gaertn. ssp. nucifera), one of 12 aquatic species used as vegetable, has been cultivated for more than 2,000 years, and now has been widely cultivated in almost all provinces in China. The largest area under cultivation of lotus is located in the regions surrounding mid-down Yangtse River, including Hubei, Jiangsu, Zhejiang, Anhui, Jiangxi and Hunan provinces. According to different purposes or morphological differences, the Chinese lotus (N. nucifera ssp. nucifera) is usually classified into three types: rhizome lotus, seed lotus and flower lotus. Rhizome lotus is mainly cultivated in Hubei, Jiangsu, Anhui and Zhejiang provinces; Seed lotus in Jiangxi, Fujian and Hunan, and flower lotus in Wuhan, Hubei province, and Beijing. Up to the year 2002, a total of 572 lotus accessions (including landraces, cultivars and breeding lines) were conserved in National Garden of Aquatic Vegetable, Wuhan, Hubei province, including those collections from 153 counties in 18 provinces, and lines bred by breeders. Out of these accessions, 310 were rhizome lotus which contains 201 landraces and 109 breeding lines; 229 were flower lotus including 172 cultivars and 57 breeding lines; and the rest 33 were seed lotus with 18 cultivars.     

Genetic diversity of a <Emphasis Type="Italic">Coffea</Emphasis> Germplasm Collection assessed by RAPD markers

Genetic diversity and relationships within and among nine species of Coffea, one species of Psilanthus and the Piatã hybrid from the Coffee Germplasm Collection of Instituto Agronômico de Campinas (IAC), Brazil were assessed using RAPD markers. Genetic diversity and relationships were evaluated by proportion of polymorphic loci (P), Shannon’s genetic index (H′ and G′_ST) and clustering analysis. The overall RAPD variation among all accessions was mostly partitioned between rather than within species. However, C. canephora and C. liberica showed a high genetic diversity within the species (\({\underline{\hbox{H}'}} \) _sp = 0.414 and \({\underline{\hbox{H}'}} \) _sp = 0.380, respectively) and this was highly structured (high \({\underline{\hbox{G}'}} \) _ST). Genetic diversity from C. congensis and C. arabica was also structured, but with lower levels of genetic diversity (\({\underline{\hbox{H}'}} \) _sp = 0.218 and \({\underline{\hbox{H}'}} \) _sp = 0.126, respectively). The results were consistent with agronomic and molecular studies and demonstrated that the IAC Coffea Collection is representative of the phylogenetic structure observed in the genera. This study devises sampling strategies for coffee germplasm collections and provides genetic diversity parameters for future comparisons among them.     

Genetic structure and history of Swiss maize (<Emphasis Type="Italic">Zea mays</Emphasis> L. ssp. <Emphasis Type="Italic">mays</Emphasis>) landraces

Between 1930 and 2003 with emphasis on the 1940s maize landraces (Zea mays L. ssp. mays) from all over Switzerland were collected for maintenance and further use in a new Swiss breeding program. The genetic relationship and diversity among these accessions stored in the Swiss gene bank is largely unknown. Our hypothesis was that due to the unique geographic, climatic, and cultural diversity in Switzerland a diverse population of maize landraces had developed over the past three centuries. The aims were to characterize the genetic diversity of the Swiss landraces and their genetic relationship with accessions from neighbouring regions as well as reviewing their history, collection, and maintenance. The characterization and grouping was based on analyses with ten microsatellite markers. Geographic, cultural, and climatic conditions explained a division in two distinct groups of accessions. One group consisted of landraces collected in the southern parts of Switzerland. This group was related to the Italian Orange Flints. The other group contained accessions from northern Switzerland which were related to Northern European Flints in particular German Flints. Historic evidence was found for a frequent exchange of landraces within the country resulting in a lack of region-specific or landrace-specific genetic groups. The relatively large separation between the accessions, indicated by high F _ST (0.42), might be explained partly by a bottleneck during the collection and maintenance phase as well as by geographical and cultural separation of north and south of the country. Due to the high genetic diversity, the accessions here are a potential resource for broadening the European flint pool.     

Genetic diversity of Swiss maize (<Emphasis Type="Italic">Zea mays</Emphasis> L. ssp. <Emphasis Type="Italic">mays</Emphasis>) assessed with individuals and bulks on agarose gels

About 65 years ago, more than 150 Swiss maize landraces (Zea mays L. ssp. mays) of the flint type were collected and conserved ex situ. Due to the climatically and culturally diverse environment of the Alps, a considerable genetic diversity of this material was assumed. To prove this, an efficient method was required to carry out genetic profiling of all the accessions in the Swiss Gene Bank. Simple sequence repeat marker (SSR) profiling in combination with the visualization of the polymerase chain reaction (PCR) products on agarose gels was chosen. Here a set of 19 different landrace accessions was analyzed to: (i) investigate their genetic diversity, (ii) investigate and display the population structure and (iii) determine whether DNA bulks rather than single plants can be used for such analyses. Four repeated samples of one accession were found to be much closer to one another than to the rest of accessions. Furthermore, specific alleles were identified for several accessions. The PCR products of the bulked DNA samples represented only a small part of the variation revealed by the analysis of individuals. Loci with four base repeat motifs performed better in the analysis of bulks than loci with other repeat motifs. The correlation between genetic distance matrices, based on the analysis of individuals and bulks, respectively, was significant. Thus, the single plant approach allowed for sufficient differentiation of accessions, and DNA bulks visualized on agarose gels led to correlated genetic distances although a limited number of alleles were detected. Although the limited resolution of agarose gels likely causes some bias, profiling of larger sets with the individual plant approach appears feasible and more informative compared to the bulk analysis we conducted.