Genetic Diversity in a Worldwide Collection of Cherimoya Cultivars

Genetic diversity of 206 cherimoya (Annona cherimola Mill.) cultivars representing the major regions of production was determined from an analysis of 23 isozyme loci. Gene diversity in cherimoya was high, although not equally distributed among countries. Peru and Ecuador, the countries of origin of this species, showed the greatest diversity, and Spain and Madeira (Portugal) showed the least, probably due to genetics bottlenecks resulting from limited introduction. Cultivars from California showed high values for several genetic diversity measures, likely due to their diverse origins. The expansion of cherimoya cultivation out the area of origin has produced a change in the apportionment of genetic diversity, with an increase in the inter-populational components, but likely without a general erosion overall.     

Pattern of genetic variability of <Emphasis Type="Italic">Solanum habrochaites</Emphasis> in its natural area of distribution

The tomato wild relative species Solanum habrochaites (previously known as Lycopersicon hirsutum) is a potential source of novel genes for tomato breeding. It shows resistance to many diseases and pests, cold tolerance and fruit quality traits. This species inhabits the western Andean slopes at high elevations from central Ecuador to central Peru. In this study the genetic variation of S. habrochaites was studied using 91 accessions from the whole range of distribution of this species. To this end, we employed two kinds of markers: 9 SSRs and 6 AFLP combinations. The Principal Coordinate Analysis based on AFLP data showed the existence of clinal genetic variation from north to south. The accessions of the different geographic groups were sequentially arranged in the first axis from north to south and a clear separation between them was found. The groups from the centre of the area of distribution showed the highest variation and heterozygosis. The ones from the margins showed lower variability and presented higher homozygosis. The morphotypes typicum and glabratum considered by Müller constituted the extreme forms of the continuous variation in the pubescence grade. These differences in pubescence were not associated with the homozygosis grade.     

Genetic and bioclimatic variation in <Emphasis Type="Italic">Solanum pimpinellifolium</Emphasis>

Solanum pimpinellifolium, due to its close relationship to S. lycopersicum, has been a genetic source for many commercially important tomato traits. It is a wild species found in the coastal areas of Peru and Ecuador. In this study, the genetic variation of S. pimpinellifolium was studied using the diversity found in 10 microsatellites in 248 plants spread throughout its entire distribution area, including Ecuador, which has been underrepresented in previous studies. Peruvian and Ecuadorian accessions are genetically quite differentiated. A possible cause of these differences could be the non-uniform nature of the coastal Ecuadorian and Peruvian climates, seeing as an important correlation between genetic differentiation and climate has been found. In addition, Ecuadorian and south Peruvian accessions have a lower genetic diversity and a higher homozygosity due to their higher autogamy, lower population size, and possible colonization bottlenecks. The Galápagos Islands population is an extreme case, with no diversity, likely caused by a recent colonization from the northern continental Ecuadorian region where genetically identical plants have been found. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Elena Zuriaga and José M. Blanca contributed equally to this work.     

Intra-specific DNA polymorphism in pineapple (<Emphasis Type="Italic">Ananas comosus</Emphasis> (L.) Merr.) assessed by AFLP markers

Pineapple (Ananas comosus (L.) Merr.) cultivars, often derived from somatic mutations, are propagated vegetatively. It has been suggested by isozyme data that there is little genetic variation among Smooth Cayenne cultivars. A thorough investigation of the genetic variation within the cultivated speciesAnanas comosus, particularly among commercial cultivars, will provide critical information needed for crop improvement and cultivar protection. One-hundred and forty-eight accessions ofA. comosus and 14 accessions of related species were evaluated with AFLP markers. The average genetic similarity ofA. comosus was 0.735 ranging from 0.549 to 0.972, suggesting a high degree of genetic variation within this species. With AFLP markers, discrete DNA fingerprints were detected for each commercial cultivar, breeding line, and intra-specific hybrid. Self-incompatibility, high levels of somatic mutation, and intraspecific hybridization may account for this high degree of variation. However, major cultivar groups of pineapple, such as Cayenne, Spanish, and Queen, could not be distinctively separated. These cultivar groups are based on morphological similarity, and the similar appearance can be caused by a few mutations that occurred on different genetic background. Our results suggest that there is abundant genetic variation within existing pineapple germplasm for selection, and discrete DNA fingerprinting patterns for commercial cultivars can be detected for cultivar protection. The genetic diversity and relationships of fourAnanas species are also discussed.     

PCR Marker-based Analysis of Wild and Cultivated Yams (<Emphasis Type="Italic">Dioscorea</Emphasis> spp.) in Nigeria: Genetic Relationships and Implications for <Emphasis Type="Italic">ex situ</Emphasis> Conservation

Reliable characterization of the variation among wild and cultivated yams in Nigeria is essential for improved management and efficient utilization of yam genetic resources. RAPD and double stringency PCR (DS-PCR) analyses were used to investigate genetic relationships and the extent of redundancy among 30 accessions of two cultivated, and 35 accessions of four wild yam species collected from Nigeria. Twenty-five selected random decamer and two microsatellite primers were used individually and in combination to generate DNA profiles for each accession of the six Dioscorea species. The number of amplified fragments varied from 7 to 18 fragments per primer/primer combination. Different levels of intraspecific genetic diversity were found, with Dioscorea rotundata Poir. being the most variable. Based on identical profiles for the RAPD and DS-PCR primers, 12 duplication groups consisting of a total number of 37 accessions were observed in the present study. An UPGMA analysis grouped the majority of plants according to the species. Cultivated yams belonging to the D. cayenensis–rotundata species complex, which were classified into seven morphotypes/varietal groups, could be clearly separated into two major groups corresponding to D. rotundata Poir. and D. cayenensis Lam. D. cayenensis cultivars exhibited a low level of intraspecific variation and were genetically close to the wild species Dioscorea burkilliana J. Miège. D. rotundata cultivars classified into six varietal groups showed a high degree of DNA polymorphism and were separated into two major groups that appeared most closely related to Dioscorea praehensilis Benth. and Dioscorea liebrechtsiana de Wild. We propose, based on these results, that cultivars classified into D. cayenensis should be considered as a taxon separate from D. rotundata. The implications of intraspecific variability for the ex situ conservation of wild and cultivated yam germplasm in Nigeria are discussed.     

Genetic diversity of <Emphasis Type="Italic">Nelumbo</Emphasis> accessions revealed by RAPD

Total 65 lotus accessions in genus Nelumbo mainly collected from China, were subjected to random amplified polymorphic DNA (RAPD) markers to estimate the genetic diversity and to test the genetic basis of the relationships between morphotypes and molecular markers. Seventeen primers generated a total of 195 highly reproducible and discernible loci, among which 173 were polymorphic. Percent polymorphism varied from 66.7 to 100 with an average of 88.72, and five primers out of them, OPC05, OPG10, OPN20, OPP09 and OPS17, showed 100% polymorphism. A relatively high genetic diversity was detected among all the samples with the similarity coefficient values ranging from 0.45 to 0.85, and Nei’s gene diversity (h) 0.30, and Shannon index (I) 0.46. The UPGMA dendrogram clustered 65 accessions in four clusters and the clustering pattern showed two groups, N. nucifera ssp. nucifera and those accessions related to the American lotus, and some special cultivars, landraces, hybrids and the American lotus. Principal Coordinate Analysis (PCA) further indicated that the genetic diversity of Nelumbo accessions was not evenly distributed, instead, was presented by a clustered distribution pattern. Similar to the results revealed by the dendrogram, two main groups representing the two subspecies of N. nucifera, as well as some special landraces, cultivars of Chinese lotus, the Japanese lotus and hybrids out of the two groups were obtained. Neither the UPGMA dendrogram nor the PCA analysis exhibited strict relationship with geographic distribution and morphotypes among the accessions.     

Variability in sesquiterpene lactones from the leaves of yacon (Smallanthus sonchifolius) accessions of different geographic origin

The sesquiterpene lactones (STLs) content of glandular trichomes from the leaves of twenty-five yacon (Smallanthus sonchifolius, Asteraceae) accessions, obtained along a latitudinal gradient from Ecuador to northwest Argentina, was characterized by gas chromatography/mass spectroscopy (GC/MS). While accessions from Ecuador, Bolivia and Argentina proved to be very chemoconsistent, significant variation was found in quantitative composition of STLs from accessions in central Peru, the probable region of origin for the species.     

Genetic evidence for speciation in <Emphasis Type="Italic">Cannabis</Emphasis> (Cannabaceae)

Sample populations of 157 Cannabis accessions of diverse geographic origin were surveyed for allozyme variation at 17 gene loci. The frequencies of 52 alleles were subjected to principal components analysis. A scatter plot revealed two major groups of accessions. The sativa gene pool includes fiber/seed landraces from Europe, Asia Minor, and Central Asia, and ruderal populations from Eastern Europe. The indica gene pool includes fiber/seed landraces from eastern Asia, narrow-leafleted drug strains from southern Asia, Africa, and Latin America, wide-leafleted drug strains from Afghanistan and Pakistan, and feral populations from India and Nepal. A third putative gene pool includes ruderal populations from Central Asia. None of the previous taxonomic concepts that were tested adequately circumscribe the sativa and indica gene pools. A polytypic concept of Cannabis is proposed, which recognizes three species, C. sativa, C. indica and C. ruderalis, and seven putative taxa.     

Genetic Diversity Among Brazilian Cultivars and Landraces of Tomato Lycopersicon Esculentum Mill. Revealed by RAPD Markers

Random amplified polymorphic DNA markers (RAPD) were used to estimate the variability of 35 tomato accessions (Lycopersicon esculentum Mill.). A total of 257 reproducibly scorable bands were obtained from 20 primers, 78.6% of which were polymorphic. The percentage distribution of RAPD markers shows a bimodal distribution, and the frequency of rare alleles is similar in commercial and landrace accessions. Genetic distances among accessions were calculated and a dendrogram showing the genetic relationships among them was constructed allowing for the separation of four groups. Twenty out of 23 Brazilian landraces fell within one group, whereas commercial cultivars were distributed in the four groups. AMOVA analysis of RAPD data showed that, despite the high within Brazilian landraces and commercial cultivars variation, these two groups are significantly different, indicating that landraces can be a source of variation for breeding programs.     

Isozyme Variation in <Emphasis Type="Italic">Passiflora</Emphasis> Subgenus <Emphasis Type="Italic">Tacsonia</Emphasis>: Geographic and Interspecific Differentiation Among the Three Most Common Species

In a general study of banana passion fruit genetic resources, diversity was analyzed in the two main cultigens, P. tripartita var. mollissima and P. tarminiana, and their closest wild relative, P. mixta, scoring isozyme bands (IDH,PGM,ACP,PGD,DIA,andPRX) on288 plants from 31 accessions. Polymorphismandallelic richness, Nei diversity indices, and neighbor joining clustering showed that variation was poor in the cultigens in northern and central Colombia, while P. mixta appeared much more polymorphic. The populations of P. tripartita var. mollissima and P. mixta from southern Colombia and Ecuador show higher diversity values and are clearly differentiated from those of central and northern Colombia. This geographic component of variation is even stronger than the interspecific one, which suggests a close relation and a regular gene flow between these two species. In contrast, all the accessions of P. tarminiana constitute a clearly differentiated group, even if some introgression with P. tripartita var. mollissima is also suspected. The high variation observed in the southern region indicates the proximity of a center of diversity for banana passion fruit and collecting in Ecuador, Peru and Bolivia is recommended. The high diversity of P. mixta and the evidence of gene flow with P. tripartita var. mollissima constitute a favorable context for the implementation of in situ conservation strategies.     

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 and structure of Ethiopian,Yemen and Brazilian C<Emphasis Type="Italic">offea arabica</Emphasis> L. accessions using microsatellites markers

Genetic diversity among 115 coffee accessions from the Coffea Germplasm Collection of IAC was assessed using SSR markers. The germplasm represents 73 accessions of Coffea arabica derived from spontaneous and subspontaneous plants in Ethiopia and Eritrea, species center of origin and diversity, 13 commercial cultivars of C. arabica developed by the Breeding Program of IAC, 1 accession of C. arabica cv. ‘Geisha’, 13 accessions of C. arabica from Yemen, 5 accessions of C. eugenioides, 4 accessions of C. racemosa and 6 accessions of C. canephora. Genetic analysis was performed using average number of alleles per locus (A), proportion of polymorphic loci (P), Shannon’s genetic index (H′ and G′_ST) and clustering analysis. All evaluated species were distinguished by a cluster analysis based on Jaccard’s coefficient. Differentiation between the cultivated plants of C. arabica and accessions derived from spontaneous and subspontaneous plants was observed. Spontaneous and subspontaneous accessions from Ethiopia were separated according to the geographical origin: east and west of the Great Rift Valley. Cultivated plants showed a low genetic diversity with a division in two groups: accessions from Yemen (H′=0,028) and Brazilian commercial cultivars (H′=0,030). The results agreed with previously reported narrow genetic basis of cultivated plants of C. arabica and supported the hypotheses about domestication of the species. This study also showed a significant genetic diversity among accessions from Ethiopia and Eritrea present in the Germplasm Collection of IAC. This diversity is specially observed in accessions from Sidamo (H′=0,143), Kaffa (H′=0,142) and Illubabor (H′=0,147) indicating their importance as source of genetic variability for coffee breeding programs.     

Preliminary study of genetic diversity in Swedish flax (Linum usitatissimum)

To investigate the genetic diversity of Linum usitatissimumL. in Sweden, 18 accessions, including 13 cultivars and five landraces, were analysed. This study was based on genetic variation in three enzyme systems (i.e., PGD, GPI and MDH) by using horizontal starch gel electrophoresis. The total genetic diversity of the studied flax material was very high (H _T= 0.62). Even though the highest genetic diversity lies within the accessions (G _ST= 0.07), a clear differentiation between fibre and oil flax was found with respect to three polymorphic loci (Pgd-1, Gpi-2 and Mdh-1). A phenogram, based on Nei's genetic distances between the accessions studied, showed five clearly defined groups but with low variation within the groups. The unexpected high genetic diversity found within accessions in the studied flax material may indicate that flax is more outbreeding than earlier believed.     

Diversity of <Emphasis Type="Italic">Ullucus tuberosus</Emphasis> (Basellaceae) in the Colombian Andes and notes on ulluco domestication based on morphological and molecular data

Ullucus tuberosus (common name ‘ulluco’) is a popular tuber crop of Andean highlands. Until now attention has been focused on ulluco from Bolivia, Peru and Ecuador, but little is known about its diversity in Colombia. Thirty-six accessions of cultivated ulluco preserved in the Gene Bank of the Universidad Nacional de Colombia were studied to assess the genetic diversity and spatial genetic structure of Colombian ulluco. We used morphological characters, molecular markers (total proteins, isozymes and RAPDs) and ploidy level. High morphological variability, especially for tuber shape and colour characters, was found. Eight accessions from north-east Colombian Andes showed some typical character-states of wild ulluco, suggesting that they could be partially domesticated forms. Genetic analysis using RAPD markers indicated that these accessions were very similar to other cultivated ullucos belonging to the same region. The regional structure observed in isozyme and RAPD dendrograms was confirmed by AMOVA results (52.6% among-region variation) and the spatial correlogram, showing the presence of two gene pools of ulluco in Colombia. Our results and the accumulated data suggest that ulluco was introduced to Colombia at least two times. Initially, semi-domesticated forms would have come from the central Andes to north-east Colombian Andes, where the native inhabitants would have completed the domestication process. Again, fully domesticated ullucos might have been introduced to south-west Colombian Andes.     

Processes affecting genetic structure and conservation: a case study of wild and cultivated <Emphasis Type="Italic">Brassica rapa</Emphasis>

When planning optimal conservation strategies for wild and cultivated types of a plant species, a number of influencing biological and environmental factors should be considered from the outset. In the present study Brassica rapa was used to illustrate this: to develop Scandinavian conservation strategies for wild and cultivated B. rapa, DNA-marker analysis was performed on 15 cultivated and 17 wild accessions of B. rapa plus 8 accessions of the cross compatible B. napus. The B. rapa cultivars were bred in Sweden and Finland in 1944–1997 and the wild B. rapa material was collected from Denmark, Sweden and United Kingdom. The B. napus accessions were bred within the last 20 years in the Scandinavian countries. Results were based on scoring of 131 polymorphic ISSR markers in the total plant material. A Bayesian Markov chain Monte Carlo (MCMC) approach implemented in NewHybrids demonstrated a clear distinction of B. rapa and B. napus individuals except for three individuals that seemed to be backcrosses. The backcrossed hybrids descended from two Swedish populations, one wild and one escaped. The overall pattern of genetic variation and structure in B. rapa showed that cultivated and wild B. rapa accessions formed two almost separated clusters. Geographical origin and breeding history of cultivars were reflected in these genetic relationships. In addition, wild populations from Denmark and Sweden seemed to be closely related, except for a Swedish population, which seemingly was an escaped cultivar. The study point to that many processes, e.g. spontaneous introgression, naturalisation, breeding and agricultural practise affected the genetic structure of wild and cultivated B. rapa populations.     

Genetic diversity and phylogenetic relationship of <Emphasis Type="Italic">Tadehagi</Emphasis> in southwest China evaluated by inter-simple sequence repeat (ISSR)

There are many valuable Tadehagi accessions in southwest China, but it is unknown that the genetic diversity and phylogenetic relationship of these Tadehagi resources. This report is the first study in which 41 primers of inter-simple sequence repeat (ISSR) were used to assess the genetic diversity of 36 Tadehagi accessions from 3 provinces in the southwest of China. Totally, 30 usable ISSR primers detected 163 polymorphic bands among the 36 accessions, which suggested high utility of ISSR primers in the genetic analysis of Tadehagi accessions. Genetic similarity coefficients among all of the accessions ranged from 0.54 to 0.92 with the average of 0.79 based on the ISSR data, indicating high level of genetic variation in Tadehagi resources from the southwest of China. As for the 3 population, Hainan population had the maximum average genetic similarity coefficients of 0.81, while similarity coefficient of Guangxi and Yunnan population was 0.75 and 0.74, respectively. All the 36 Tadehagi accessions were divided into 4 groups in the UPGMA dendrogram constructed from genetic similarity coefficients. The Tadehagi accessions from Yunnan and Guangxi provinces showed more genetic variation and occupied the bottom of the dendrogram. On the contrary, those from Hainan Province had less genetic variation and clustered in the middle and top of the dendrogram. The information on the genetic diversity and phylogenetic relationship from this study is propitious to construct a core germplasm collection and develop novel Tadehagi cultivars with desired economic traits.     

Assessments of genetic diversity within a world collection of cultivated hexaploid oat (<Emphasis Type="Italic">Avena sativa</Emphasis> L.) based on qualitative morphological characters

A characterization of 10,105 accessions of hexaploid cultivated oat (Avena sativa L. sensu lato) from 85 countries of the Plant Gene Resources of Canada (PGRC) collection was conducted at Saskatoon, Saskatchewan, Canada. Eight environmentally stable morphological characters (panicle type, panicle erectness, panicle density, lemma colour, dorsal awn of lemma, ligula, major infraspecific group), differentiated into 18 character states, were used to define genetically distinct morphological groups. Comparisons of oat diversity from different countries, among the major infraspecific groups and among Canadian oat cultivars registered between 1886 and 2002 were possible. The 10,105 accessions represented 118 different morphological groups. The number of accessions in each morphological group was unevenly distributed with the 13 most frequent morphological groups including 90% of the accessions. The most frequent morphological groups in the PGRC collection were identical with the most frequent types of Canadian oat cultivars. The greatest richness of diversity was found in oat from countries with temperate climates and intensive oat breeding programmes. The oat accessions comprised 8,754 accessions of common hulled oat, 183 accessions of hull-less oat and 1,168 accessions of red oat. For red oat (A. byzantina C. Koch), West Asia was richest in diversity. The USA could be considered a secondary centre of diversity for red oat and Canada a secondary centre of diversity for hull-less oat. Morphological diversity of oat cultivars released in Canada increased during the twentieth century. The morphological groups were related to formal taxonomical infraspecific classifications of A. sativa. Applications of the concept of defining morphological groups for phenotyping a large germplasm collection are demonstrated discussed.     

Imbibition behavior and flooding tolerance of rapeseed seed (<Emphasis Type="Italic">Brassica napus</Emphasis> L.) with different testa color

In this study, the correlations among these characters were investigated in 18 rapeseed (Brassica napus L.) accessions with different seed coat color. The results indicated that seed water uptake and flooding tolerance were significantly correlated with seed color and melanin pigment content of testa. The red or black-seeded accessions had higher melanin content in testa, showed slower water uptake and lower leakage and higher flooding tolerance. The majority of yellow-seeded rapeseed accessions which had low melanin pigments content in testa showed a rapid water uptake and higher leakage than the red or black-seeded, which led to imbibition damage and lower flooding tolerance. The results suggest that the yellow-seeded cultivars experienced poor field emergence and more serious pre-harvest sprouting in raining weather than the dark-seeded cultivars. Some yellow-seeded accessions showed a slow imbibition behavior and relative high flooding tolerance, indicated these accessions can be used as a genetic resource to improve the flooding tolerance and reduce imbibition damage for the yellow-seeded B. napus L.     

Application of random amplified polymorphic DNA to study genetic diversity in Paspalum scrobiculatum L. (Kodo millet, Poaceae)

Summary Genetic diversity and patterns of geographic variation among collections of Paspalum scrobiculatum (kodo millet) and P. polystachyum were studied using molecular markers generated through the random amplified polymorphic DNA (RAPD) method. A high level of polymorphism in RAPD markers was observed among the individual accessions, demonstrating the high genetic diversity of the crop. The markers obtained from the RAPD method were analyzed with the cluster analysis, principal coordinates and minimum spanning tree methods. Three major groups were resolved, one representing the African accessions, and two for the Indian accessions. The accessions of the north African kodo millet and P. polystachyum (considered conspecific with P. scrobiculatum) were quite distinct. The Australian kodo millet showed higher affinity to the African types. The study demonstrated that the RAPD technique can be applied to resolving degrees and patterns of genetic variation at the population and species levels, identifying cultivars, and defining gene pools of this crop.