Spatial distribution of nutrients and morpho-physiological indicators of salinity tolerance among five olive cultivars - The use of relative nutrient concentration as an efficient tolerance index

Abstract

Olive is the most important fruit tree species grown in Mediterranean basin, where many times it grows under saline conditions, due to irrigation with low quality water. Olive response to salinity is cultivar dependent and the evaluation of native genotypes is an important tool in finding tolerant genetic material. For this reason, five Greek olive cultivars, i.e. “Koroneiki”, “Gaidourelia”, “Lefkolia Serron”, “N-K Gigas” and “Throumbolia”, were evaluated regarding their tolerance to sodium chloride salinity (0, 50, 100 and 200?mM). All cultivars exhibited a significant reduction of growth, while the most severe symptoms were found in “Gaidourelia” while “Lefkolia Serron” presented the least symptoms. The latter exhibited the lowest sodium concentration in the leaves and the highest in the roots. Both potassium to sodium and calcium to sodium ratios were high in “Gaidourelia” leaves and stems, suggesting that the use of nutrient concentration for comparison of olive tolerance among cultivars could lead to erroneous results. The best indicator proved to be the determination of nutrient relative concentration compared to corresponding cultivar’s control, as “Gaidourelia” exhibited the highest sodium relative concentration as well as the lowest nutrient ratios, revealing its inability to restrain sodium in the root level.     

Genetic diversity in local cultivars of garden pea (Pisum sativum L.) conserved ‘on farm’ and in historical collections

During a national Swedish collection mission of vegetable varieties conserved ‘on farm’ more than 70 pea accessions were obtained, many of which had been grown locally for more than 100 years. In spite of a likely origin in the multitude of obsolete commercial pea varieties available on the Swedish seed market in the nineteenth century, the rediscovered local cultivars have lost their original names and cultivar identity while being maintained ‘on farm’. To analyze genetic diversity in the repatriated material, 20 accessions were genotyped with twelve SSR markers and compared with 15 obsolete cultivars kept in genebanks and 13 cultivars preserved as non-viable seeds collected in 1877–1918. Most of the local cultivars were genetically distinct from each other, and in only a few cases could a possible origin in a tested obsolete cultivar be suggested. These results reflect the wide diversity of pea cultivars present in Sweden during the nineteenth century. Both between and within accession genetic diversity was larger among the historical samples of obsolete cultivars compared to local cultivars and cultivars preserved in genebanks, indicating genetic erosion over time both in genebanks and during conservation ‘on farm’. The constraints on identifying and verifying historical cultivars using genetic markers are discussed.     

Genetic diversity and relationships between wild and cultivated olives (<Emphasis Type="Italic">Olea europaea</Emphasis> L.) in Sardinia as assessed by SSR markers

The genetic relationships within and between wild and cultivated olives were examined and clarified in an isolated and restricted area, such as the Mediterranean island of Sardinia. Wild (21 individuals) and cultivated olive trees (22 local cultivars from a germplasm collection and 35 ancient trees) were genotyped by means of 13 SSR loci. Five cases of synonymy were observed and nine distinct genotypes were identified in the collection. Five novel genotypes were also detected among the ancient trees. Differences on the allelic composition and heterozygosity levels were found between wild and cultivated trees. Model-based clustering method classified the olive trees into two major gene pools: (a) wild genotypes and (b) local cultivars from the collection and from heritage olives. Regarding the cultivated plant material, we observed that: (a) most of the Sardinian cultivars shared the same allelic profiles with the ancient cultivated trees and (b) the majority of these cultivars and all the novel genotypes were not related to any other cultivars included in this study. These findings as well as the detection of unique alleles and a certain wild genetic background at some cultivars revealed by the Bayesian analysis may indicate their autochthonous origin. The synonymy cases found between local cultivars and Italian mainland cultivars indicate interchange of genetic material among these growing areas, suggesting thus a possible allochthonous origin. The information obtained can assist in the management of an olive collection and sheds some light on the survival of true oleasters and the origin of Sardinian cultivars.     

Genetic relationships among local potato cultivars from Spain using SSR markers

Microsatellite markers (SSR) were used to fingerprint a total of 105 local potato cultivars from Spain. A set of 41 cultivars from Tenerife Island, 19 from the island of La Palma, and 45 local varieties from peninsular Spain were analysed. Some of these varieties represent relicts of the early introductions originating from South America and have been characterised previously morphologically and ecophysiologically. We observed within our materials a total of 76 SSR alleles. Only seven of them were present in all varieties. Several accession and group specific alleles were detected. Similarity coefficients were computed from the molecular data and cluster analyses were performed. The obtained dendrogram was generally in good agreement with previous classifications of the accessions as Solanum tuberosum L. subsp. andigena (Juz. et Bukasov) Hawkes S. tuberosum L. subsp. tuberosum and S. chaucha Juz. et Bukasov genotypes. Also cultivar groups with identical or related common names showed the same SSR patterns or clustered closely together. In addition we performed Principal Coordinate Analysis with the set of genotypes. Results of both analysis methods were generally in good agreement, but also some smaller differences were detected in the associations of groups and genotypes. According to the molecular patterns for some accessions misleading or confounded names were evident, and in some cases the molecular patterns showed also discrepancies with previous species assignments, suggesting the need for a more detailed comparative study of these accessions.     

The coexistence of oleaster and traditional varieties affects genetic diversity and population structure in Algerian olive (<Emphasis Type="Italic">Olea europaea</Emphasis>) germplasm

The present work was aimed at assessing the genetic diversity of 42 local cultivars and oleaster genotypes from the area of Bejaia in Algeria. Fifteen highly polymorphic Simple Sequence Repeat markers were evaluated and proved to be very informative, producing a total number of 160 alleles with an average value of 10.7 per locus; the SSRs DCA09 and DCA16 were the most informative, distinguishing 17 and 19 genotypes, respectively. Phylogenetic and population structure analysis split the accessions in two main groups corresponding to most of oleasters and most of traditional varieties, respectively. Interestingly, ten traditional varieties resulted strictly related to the oleasters, indicating hybridization between the two botanical varieties. Genetic parameters and private alleles of groups confirmed this observation and indicated a wide genetic variability in Algerian olive germplasm. The results suggest the need to preserve and characterize this germplasm in order to limit the risk of losing potential important genetic traits present in the crop wild relatives.     

Pigment metabolism of 'Sikitita' olive ( Olea europaea L.): a new cultivar obtained by cross-breeding

The new olive cultivar 'Sikitita' was obtained from a cross between the 'Picual' and 'Arbequina' varieties. 'Sikitita' was selected for its features, making it particularly suited to high-density olive hedgerow orchards. From the standpoint of chloroplast pigment metabolism, the fruits of the 'Picual' and 'Arbequina' varieties have significant differences. It is therefore extremely interesting to analyze the descendants of both cultivars. With regard to chlorophyll catabolism, 'Sikitita' has proven to be a cultivar with low pigmentation and low levels of chlorophyllase activity. This is contrary to the findings obtained to date, where varieties with low pigmentation are a consequence of high chlorophyllase activity ('Arbequina') and highly pigmented fruits are due to low chlorophyllase activity ('Picual'). 'Arbequina' was, until recently, the only cultivar described that had developed a carotenogenic process, despite its anthocyanic ripening. However, from its father ('Arbequina'), the 'Sikitita' cultivar has inherited the pool of enzymes necessary to esterify xanthophylls at the chromoplast level. This makes 'Sikitita' a very interesting cultivar, with potential chemotaxonomic differences (such as esterified xanthophylls in the olive oils), and demonstrates the interest in genetic improvement programs for olive cultivars with different organoleptic characteristics.     

Menara gardens: a Moroccan olive germplasm collection identified by a SSR locus-based genetic study

The Menara gardens in Marrakech (Morocco), established in the 12th century, comprise more than 2000 olive trees. To assess the genetic diversity within these gardens, we performed SSR analysis of 128 randomly sampled olive trees. Using 15 SSR loci which revealed 70 alleles, we identified 16 distinct genotypes. The analysis of chloroplast DNA polymorphism allowed to identify 2 chlorotypes: COM1 for 2 genotypes and CE1 for all others. Among the 128 trees analysed, Picholine marocaine was the prevalent cultivar represented by 100 trees (78%), the 28 remaining trees being classified into 15 distinct genotypes. However, four genotypes were closely related to Picholine marocaine from which they probably derived through somatic mutations. The genotypic and genetic diversity observed in Menara gardens compared with the genotype diversity from different areas (North and Atlas); these facts support the hypothesis that these gardens have been planted with trees probably originating from different areas of Morocco. Our results point out an important concept for ex situ conservation: the assumption that local Moroccan olive germplasm was empirically, but not purposively, conserved already in Menara gardens.     

Exploring genetic variation,oil and α-tocopherol content in avocado (<Emphasis Type="Italic">Persea americana</Emphasis>) from northwestern Mexico

The Hass cultivar of avocado is the most widely grown commercial cultivar in Mexico. Unfortunately, this cultivar is poorly adapted to the Mexican low-lands with hot dry climates characteristic of northwestern Mexico. Other well-adapted avocado accessions are available for these regions, but their nutritional traits and genetic diversity have yet to be explored. In this study, we analyze oil content, (α-tocopherol) and genetic variation among five local varieties from northwest Mexico that grow in high temperatures regimes in unfertilized soils and without any agronomic management. We report significant phenotypic variability in oil and α-tocopherol components in different accessions of avocado as determined by HPLC. Interestingly, we find higher α-tocopherol content (45.02–50.66 μg/g of fresh pulp) in some local avocados compared to Hass (32.28 μg/g of fresh pulp). The analyzed accessions represent a moderately polymorphic set of genotypes as measured by microsatellite (10 alleles by locus) and SNP (1 SNP every 164.4 bp) analysis of the VTE3 and VTE4 genes, implied in the biosynthesis of tocopherols. SNP data allowed also identifying differences between the local varieties and controls (Hass and the Mexican race accession). The variation observed at the genetic, morphologic and nutritional levels provide significant new information that may be valuable in selecting and developing avocado genotypes adapted to high-temperature environments.     

Effect of the maturation process of the olive fruit on the phenolic fraction of drupes and oils from Arbequina, Farga, and Morrut cultivars

The purpose of the work was to investigate the effect of the maturation process of the olive fruit on the phenolic fraction of drupes and oils from Arbequina, Farga, and Morrut cultivars. The level in the phenolic content of olive drupes declines rapidly during the black maturation phase. A general decreasing trend was observed too in the phenolic content of olive oils during the ripening process in the three varieties studied. Important differences in the high-performance liquid chromatography profile between varieties were observed. These included the presence of very low amounts of lignans in olive oils proceeding from the Morrut cultivar, and the presence of three peaks after elution of 3,4-DHPEA-EDA in the Farga and Morrut cultivars, which could be used as differentiating parameters. Sensory profile differences were observed between olive cultivars and due to the ripening process.     

Identification and genetic structure of wild Italian <Emphasis Type="Italic">Humulus lupulus</Emphasis> L. and comparison with European and American hop cultivars using nuclear microsatellite markers

Nine genic SSR loci were used to evaluate the genetic diversity and identify accessions in wild Italian Humulus lupulus L., in comparison with widely cultivated European and U.S. commercial cultivars. A collection of 80 wild hop samples from Italy and 43 hop cultivars from Europe and U.S., were characterized. Allelic frequency analysis revealed 65 distinct Italian genotypes and differentiated all the commercial cultivars; moreover, specific alleles were observed for wild and cultivated hops. The number of alleles identified in the wild population were 104 and 123 within all the accessions. The maximum polymorphic information content was evidenced for locus HlGA23 in the Italian wild population and in the whole set of accessions (0.905 and 0.902 respectively). The dendrogram constructed from Euclidean distance with the UPGMA method showed two main clusters, one including commercial American and European accessions and one mostly composed by wild Italian accessions. Model-based clustering (Bayesian method) placed the accessions into five germplasm groups, one of which was characterized by Italian genotypes only. The study showed for the first time the great biodiversity present in Italy, and the remarkable differences with European and American hops. It was also found that within the population of north-central Italy a large genetic variability is present, suited to be studied and exploited; this genetic wealth could be used in future breeding programs in order to develop new hop varieties carrying characteristics useful for brewers.     

SSR Diversity of Vegetable Soybean [<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merr.]

Edamame [Glycine max (L.) Merr.] is a type of soybean selected for fresh or frozen vegetable use at an immature stage. Since edamame has a similar protein content, milder flavor, nuttier texture, and is easier to cook when compared to grain soybean, it is being promoted as a new vegetable for global consumption. Global production will require breeding programs for local adaptation; however, limited research has been published on genetic diversity of edamame varieties for the assessment of genetic resources. Simple sequence repeats (SSRs) were used to study the genetic diversity among 130 accessions, including edamame cultivars and landraces from Japan, China and the US, and also the new breeding lines in the US. Although it is assumed that elite edamame cultivars would have narrow genetic diversity, seventeen SSRs detected polymorphism to distinguish 99 of the 130 accessions. The cluster analysis generated nine clusters and 18 outliers. Genetic diversity within Japanese edamame was lower than that within Chinese vegetable soybean accessions (maodou), even though only 10 Chinese maodou were analyzed compared to 107 Japanese edamame. Cluster analysis revealed that the patterns of SSR diversity in edamame can generally distinguish maturity classes and testa color. We concluded that Japanese edamame have a narrow genetic base different from others and that SSRs can describe the patterns of genetic diversity among the elite vegetable soybean.     

ISSR Variation in Olive-tree Cultivars from Morocco and other Western Countries of the Mediterranean Basin

In olive tree (Olea europaea L.), 12 varieties (or cultivars) representing the main domesticated material used in Morocco and 19 olive cultivars used extensively in five countries of the western Mediterranean Basin, were analysed using inter-simple sequences repeat (ISSR) markers which had never been used previously for extensive discrimination of cultivars. Four selected primers produced a total of 26 polymorphic reproducible amplification fragments. Combinations of these ISSR markers allowed to identify 25 of the 31 cultivars. Two additional combinations were distinguished, each corresponding to three Moroccan cultivars or local varieties. Evidence of a multiclonal composition in the widely cultivated variety ‘Picholine marocaine’ was obtained by the identification of three genotypes within the four morphologically distinct clones analysed in the variety. In the UPGMA phenogram based on the proportion of shared ISSR fragments, five groups of cultivars were distinguished at the 40% critical value of similarity. Four of the groups contained varieties from various geographic origins, as the consequence of successive human migrations which favoured olive dispersion throughout the Mediterranean Basin. However, the fifth group gathered together 9 of the 12 Moroccan cultivars and very few cultivated clones from Greece and Spain. The results suggest that most of the Moroccan cultivars are closely related and likely originated from local domestication.     

Analysis of diversity in the fruit tree genetic resources from a Mediterranean island

A large number of genotypes of different fruit tree species have been described in Sardinia, where the plant species have evolved by adaptation to the Mediterranean environment. In this work results are summarized of a long-term survey carried out in the island by the authors since the early 1980s. Ten fruit tree species have been surveyed and collections of the traditional varieties were established. Some are major staple crops such as olive and grape, others such as almond, cherry, plum, fig, apple and pear are nowadays less cultivated, and still others have been more recently introduced like apricot and cactus pear. Data on the level of inter and intra-species diversity are given, as illustrated by representative morphological traits. The results of isozymes and RAPD-PCR analysis on the species Ficus carica genotypes are included, to assess genetic relationship among accessions. Observations on composition, as well as on state of conservation of selected traditional varieties are reported, and the main causes of genetic erosion are listed.     

RAPD and SSR markers for characterization and identification of ancient cultivars of <Emphasis Type="Italic">Olea europaea</Emphasis> L. in the Emilia region,Northern Italy

The Emilia region (Northern Italy) is characterised by the occurrence of microclimates that permit olive growing. The presence of the species, albeit sporadic, in these territories for several centuries as a fruit crop is well documented, by both archaeological and written testimony, and by a large number of plants well over a century old, located in particular sites, favourable for growth and development of the tree. Olive genetic diversity was studied using RAPD and SSR techniques, on plants growing in the Emilia territory (Reggio Emilia and Parma provinces). For genotype identification comparisons were made with 8 cultivars, some of which from Central Italy. Screening was obtained analysing patterns produced by 20 RAPD primers and 3 SSR primers, developed by other authors; the primers and we were able to discriminate olive cultivars with a sufficient degree of reliability. The dendrograms obtained from the analysis show the genetic relationship among accessions present in the Parma-Reggio Emilia district. Our results demonstrated the reliability of RAPDs and SSRs to identify all studied olive cultivars and to reveal the degree of their relatedness to each other. The analysis also reveals the presence of an interesting amount of genetic diversity among the studied individuals.     

A wide range of genetic diversity in pear (<Emphasis Type="Italic">Pyrus ussuriensis</Emphasis> var. <Emphasis Type="Italic">aromatica</Emphasis>) genetic resources from Iwate,Japan revealed by SSR and chloroplast DNA markers

Iwateyamanashi (Pyrus ussuriensis var. aromatica) is one of the Pyrus species which grows wild in Japan. The number of Iwateyamanashi trees has been decreasing, so conservation and evaluation is urgently needed. Over 500 accessions of Pyrus species collected from Iwate in northern Tohoku region are maintained at Kobe University as an Iwateyamanashi germplasm collection. In order to investigate the genetic diversity, five SSR (simple sequence repeat) markers, developed from Japanese and European pear were examined for 86 Pyrus individuals including 58 accessions from Iwate. These SSR loci could discriminate between all the Iwate accessions except for 10 that bear seedless fruit, as well as determine the genetic diversity in Iwateyamanashi germplasms. High levels of variation were detected in 41 alleles and the mean observed heterozygosity across 5 loci was 0.50 for the Iwate accessions. Seedless accessions sharing identical SSR genotype with the local pear variety “Iwatetanenashi” were supposed to have been propagated vegetatively via grafting. In an UPGMA phenogram, Japanese pear varieties (P. pyrifolia) were clustered into two groups with some Iwate accessions including seedless ones. Another 38 Iwate accessions were not clustered clearly, and there was no clear relationship between these accessions and geographical distribution or morphological characters. Allele frequency revealed that the Iwate accessions were genetically more divergent than the Japanese pear varieties. Most Japanese pears possessed a 219 bp deletion at a spacer region between the accD and psaI genes in the chloroplast DNA (cpDNA), but other Pyrus species and two Iwateyamanashi trees did not. In the Iwate accessions, 79.3% had a deletion type cpDNA and others had a standard type cpDNA without deletion. These results are indicative of the wide range of genetic diversity in the Iwate accessions which include Japanese pear varieties. A combination of SSR and cpDNA analyses revealed high heterogeneity in Iwateyamanashi and coexistence of Iwateyamanashi and hybrid progeny with P. pyrifolia. These could be reasons for the wide range of continuous morphological variation described previously.     

Identification of Chinese white pear cultivars using SSR markers

109 Pyrus accessions including 92 local Chinese accessions of P. bretschneideri were identified genetically using nine microsatellite loci developed from apple and pear. The nine SSR loci revealed 129 alleles in 109 pear accessions and 114 alleles in 92 Chinese white pears. Among the 92 local Chinese accessions of P. bretschneideri, 70 could be differentiated successfully except for 10 sets of synonymous or mutants. For the 92 accessions, the number of putative alleles per locus ranged from seven to 18, with an average of 12.67; the average values of observed heterozygosity and Shannon’s Information index were 0.60 and 1.85, respectively. A phenogram based on the SSR (simple sequence repeat) genotypes was obtained. The 109 accessions clustered into 11 groups based on geographical origin. The European pears and the Asian pears did not form independent two groups, but three P. communis cultivars grouped together independently. The Japanese P. pyrifolia cultivars mingled together with the Chinese P. bretschneideri cultivars, but four P. ussuriensis cultivars except for one (Jianbali) grouped together independently. The results indicated that the relationship of P. bretschneideri cultivars and P. pyrifolia cultivars was much closer than others.     

Relationships Among <Emphasis Type="Italic">Brassica napus</Emphasis> (L.) Germplasm from Spain and Great Britain as Determined by RAPD Markers

The genetic diversity and the relationships among a collection of Brassica napus L. European populations were evaluated using random amplified polymorphic DNA markers. The study included 33 accessions of B. napus collected from Galicia (northwestern Spain) and 18 British cultivars, 16 accessions of B. napus and two accessions of Brassica oleracea L. used as controls. DNA from 25 individuals per population was analyzed using 18 decamer primers. One hundred thirty-eight amplification products were scored of which 105 were polymorphic. These bands ranged in size from 350 to 2500 base pairs. Similarity coefficients and cluster analysis were computed and six groups were obtained. Cluster I was the largest and included all the landraces from northwestern Spain, except two accessions that grouped separately into Clusters III and IV, respectively. A low level of genetic variability was detected among the B. napus Spanish genotypes, while considerable diversity was present among the British ones, which grouped into three groups, two main clusters and one group formed by one accession. Cluster II included all commercial varieties grown in Great Britain whereas Cluster V grouped local varieties maintained by the growers for many years. Cluster VI was a singularity formed by one entry. British accessions of B. oleracea had the greatest dissimilarity with all the other populations and grouped separately in Clusters VII and VIII. As conclusion, B. napus landraces used in northwestern Spain as leafy-green vegetable probably have an independent origin from B. napus crops grown in other European regions. Besides, separate domestication in northwestern Spain and Great Britain for a different end use might have led to two distinct gene pools.     

Leaf Mineral Composition of Olive Varieties and Their Relation to Yield and Adaptation Ability

The amount of yield and adaptation of a cultivar to a new environment is strongly related to nutrient uptake ability. The aim of this study was to compare nutrient uptake ability of 21 local and/or standard olive varieties grown under Mediterranean climatic conditions. Elemental compositions of olive leaves were determined for two consecutive years. The highest yields were obtained from ‘Gemlik’, ‘Manzanilla’, ‘Memecik’, and ‘Hojiblanca’ varieties. Nitrogen (N), phosphorus (P), potassium (K), magnesium (Mg), and manganese (Mn) content of leaves were comparatively higher in the high-yielding year (2004) whereas iron (Fe) and copper (Cu) contents were lower. Nutrient uptake ability and usage efficiency of olive varieties were different. Mineral composition of leaves was variety dependent and different groups of varieties showed higher leaf concentration for each element. Therefore, it can be concluded that the variation in the nutrient uptake ability may be used as a criterion for adaptation of a variety to a new ecological environment.     

Genetic structure and differentiation among grapevines (<Emphasis Type="Italic">Vitis vinifera</Emphasis>) accessions from Maghreb region

Three gene pools representative of Vitis vinifera L. subsp. vinifera (=subsp. sativa Beger) growing in the Maghreb regions (North Africa) from Tunisia (44), Algeria (31) and Morocco (18) and 16 wild grape accessions (Vitis vinifera L. subsp. sylvestris (Gmelin) Beger) from Tunisia were analysed for genetic diversity and differentiation at twenty nuclear microsatellites markers distributed throughout the 19 grape chromosomes. 203 alleles with a mean number of 10.15 alleles per locus were observed in a total of 109 accessions. Genetic diversities were high in all populations with values ranging from 0.6775 (Moroccan cultivars) to 0.7254 (Tunisian cultivars). F _st pairwise values between cultivated grapevine populations were low but found to be significantly different from zero. High F _st pairwise values were shown between wild and cultivated compartments. Two parent offspring relationships, two synonyms and two clones of the same cultivar were detected. The rate of gene flow caused by vegetative dissemination of cultivated grapevine plants was not sufficient to genetically homogenise the pools of cultivars grown in different regions. The Neighbour Joining cluster analysis showed a clear separation according to geographical origins for the cultivated grapevines gene pools and revealed a high dissimilarity between cultivated and wild grapevine. However, three cultivars (Plant d’Ouchtata 1, Plant de Tabarka 3 and Plant d’Ouchtata 3) are very close to wild accessions and may result from a hybridisation between cultivated and wild accessions. The high level of differentiation between cultivated and wild accessions indicates that the cultivated accessions do not derive directly from local wild populations but could mostly correspond to imported materials introduced from others regions during historical times or derived from crossing between them.