Analysis of morphological variation in wild beet (Beta vulgaris L.) from Sicily

Summary In this paper the variation pattern of the Sicilian Beta germplasm collection is analysed. Accessions from Sicily were grown under controlled conditions and were evaluated morphometrically. Geographical distribution patterns of morphological characters are identified and the putative classification of the collection into coastal and inland populations is reassessed. Between adjacent populations significant variation was found for Petiole length, Leaf length, Leaf width and Biomass, however no one variable unambiguously reflected a geographical pattern. Overall differences between coastal and inland populations were small. Petiole length and rate of generative development were found to be important for distinguishing between these groups. Interferences are drawn on how the collection from Sicily could be rationalized to avoid excessive duplication.     

Phenotypic diversity and morphological characterization of Amygdalus L. species in Lebanon

In this paper, we report on morphological diversity of Amygdalus L. species native to Lebanon, following a countrywide survey of almond germplasm whereby a total of 149 accessions were collected throughout the country and characterized by thirteen quantitative and four qualitative traits. The results indicated that the genetic diversity of Amygdalus communis L., Amygdalus korshinskyi Hand.-Mazz., and Amygdalus orientalis Duh. in Lebanon is high. Principal component analysis revealed that nut weight, nut volume, nut width, kernel volume and shell strength had highest loading in the first component that accounted for 38.7% and 46.7% of total variation in A. communis and A. orientalis, respectively. In contrast, leaf traits were present in the second component which accounted for 18% and 23.2% of total variation in each species, respectively. No significant correlations were detected between leaf parameters and fruit traits in both species. The results indicated that quantitative leaf characters for all three species were determined by rainfall and not altitude whereby adjacent accessions located in drier areas had smaller leaf sizes than those located in more humid regions. Quantitative fruit characters did not seem to vary accordingly. Qualitative leaf traits in all three species reflected a variability which was independent of rainfall. A. communis populations showed high variability, suggesting that they could be a valuable source in almond improvement programs.     

Distribution and habitat of Spanish populations of the subtribe Hordeineae; improved views following germplasm collecting activities

A complete study was made of 20 Spanish herbaria and of the literature concerning the regional flora of Spain in order to define the geographical limits of the different genera of the subtribe Hordeineae found in Spain. Accordingly, several expeditions were undertaken to the greater part of the Spanish mainland and its islands in order to collect samples of wild populations of the genera Hordeum L., Taeniatherum Nevski and Hordelymus Jessen. This paper, which contains information garnered from both collection activities and the study of herbaria, describes the distribution and ecological characteristics of the habitats where populations of H. bulbosum, H. secalinum, H. murinum subsp. murinum, H. murinum subsp. leporinum, H. marinum subsp. marinum, H. marinum subsp. gussoneanum, Taeniatherum caput-medusae and Hordelymus europeus grow.The bioclimatic and geographical characteristics of the populations of the different genera and species from which samples were taken during collect ing expeditions are also described.     

Identification of <Emphasis Type="Italic">Aegilops</Emphasis> L. species and <Emphasis Type="Italic">Triticum aestivum</Emphasis> L. based on chloroplast DNA

The genus Aegilops L. is a very important genetic resource for the breeding of bread wheat Triticum aestivum. Therefore, an accurate and easy identification of Aegilops species is required. Traditionally, identification of Aegilops species has relied heavily on morphological characters. These characters, however, are either not variable enough among Aegilops species or too plastic to be used for identification at the species level. Molecular markers that are more stable within species, therefore, could be the alternative strategy towards an accurate identification. Since the chloroplast DNA has a lower level of evolution compared to the nuclear genome, an attempt was made in this study to investigate polymorphism in the chloroplast DNA among 21 Aegilops species (including Ae. mutica that is now known as Amblyopyrum muticum) and between the latter and T. aestivum to generate markers for the diagnosis of all targeted species. Cleaved amplified polymorphic sequence (CAPS) applied on 22 coding and non-coding chloroplast regions using 80 endonucleases and sequencing of two of those regions revealed little polymorphism between T. aestivum and the various Aegilops species examined and to a less extent was the variation among Aegilops species. Polymorphism observed among species analysed allowed the discrimination of T. aestivum and 12 Aegilops species.     

RAPD markers efficiently distinguish heterogenous populations of wild potato (Solanum)

Genetic characterization of germplasm is important for setting objective guidelines for conservation. One common problem found in genebanks is determining the value of populations with insufficient or unreliable data regarding their geographic origin. In this study, a genetic analysis based on RAPD markers was conducted to characterize a `mystery' population of Solanum sucrense, a polysomic tetraploid potato (2n=4x=48), for which adequate documentation was lacking. The comparative analysis of genetic similarities between this mystery population and each one of 30 other S. sucrense populations in the genebank revealed that all populations within this species, including the mystery population, are significantly different from being duplicates, and are therefore worthy of separate conservation. RAPD markers also distinguished the mystery population from closely related tetraploid species S. oplocense, S. gourlayi and S. tuberosum ssp. andigena, suggesting that it is also not a duplicate of a population of these species. If RAPDs can clearly differentiate populations within highly heterogeneous tetraploids like S. sucrense, they should be generally useful for determining germplasm organization within potato species.     

Genetic diversity in cucumber (Cucumis sativus L.): II. An evaluation of selected cultivars released between 1846 and 1978

Genetic variation among 155 U.S. modern and heirloom cultivars was assessed from assays of 21 polymorphic isozyme loci. Four loci (Fdp-1, Mdh-1, Mpi-1 and Pgd-1) were monomorphic. Multivariate analyses partitioned cultivars into two distinct groups: those released before 1968, and those released after 1968. Cluster analysis produced a dendrogram with 14 nodes and 28 groups. Modern U.S. and European cultivars released after 1968 differed in isozyme frequencies. Isozymic profiles clearly discriminated some cultivars with unique attributes and/or pedigrees [e.g., Windermoor Wonder (USA), Gergana (The Netherlands), Seiram (The Netherlands), Fancy Pak (USA), Dasher 2 (USA), and WI 2757 (USA)].     

Genetic diversity in wild Tunisian populations of Mentha pulegium L. (Lamiaceae)

The allozyme variation of 15 Tunisian wild populations of Mentha pulegium L. threatened by human activities (clearing, hard-grazing, ploughing, traditional uses) was surveyed by the analysis of 14 isozyme loci using horizontal gel starch electrophoresis. The species exhibited a high level of genetic variation within populations (the mean Ap = 2.20, P = 72%, Ho = 0.349 and He = 0.229), which indicates a predominately outcrossing mating system and the recruitment of new genotypes via dispersal seeds. The genetic structure analysis of the populations using F statistics indicates no inbreeding, and showed an excess of heterozygosity for few loci. The moderate differentiation of populations (F_ST = 0.110) and the low rate of gene flow between them (Nm = 2.02) might been caused by recent isolation of the populations through biotope disturbances. The value of Nei's genetic identity varied from 0.839 to 0.999 reflecting a relatively low genetic divergence between populations. Cluster analysis using UPGMA method and Nei's genetic identity values, showed that populations geographically close didn't always cluster together. However, populations within the same bioclimatic stage generally subclustered together indicating that differentiation between bioclimatic regions occurred.     

Analysis of genetic diversity in Turkish sesame (Sesamum indicum L.) populations using RAPD markers⋆

The genetic diversity of 38 cultivated populations of Sesamum indicum L. from four different regions of Turkey was estimated at the DNA level with the random amplified polymorphic DNA (RAPD) technique. Sixty-one bands were obtained for all populations 78% of which were polymorphic. Analysis of molecular variance (AMOVA) was used to investigate the genetic diversity of the populations which yielded highly significant differences among populations within regions (91.9% of the total genetic diversity). According to AMOVA and Shannon's index that were performed separately for each region, the highest value of genetic variation was observed among Northwest region populations (CV = 7.7; H₀ = 0.304) and lowest in the Southeast regions' populations (CV = 2.6; H₀ = 0.068). Nei and Li's similarity index was calculated and phylogenetic tree was established using the neighbor-joining algorithm. This phenetic analysis grouped 35 of 38 accessions in six groups leaving three highly diverse accessions outside. Wagner phylogenetic method was used to assess the phylogenetic relationships among the populations. In the majority-rule consensus tree, only 7 of the 32 forks showed above 60% occurrence. Using Principal Coordinate Analysis (PCO) of the RAPD data set, the groups were clearly separated along the first three axis. These results indicate that RAPD technique is useful for sesame systematics, and should be valuable for the maintenance of germplasm banks and the efficient choice of parents in breeding programs.     

Genetic variability in Turkmen populations of Pistacia vera L.

Seedlings of Pistacia vera L. developed from seeds of two separate populations in Turkmenistan, Kepele and Agachli, were evaluated for their growth potential and genetic polymorphism. Plant growth rate as well as random amplified polymorphic DNA (RAPD) analysis showed distinct differences between the two populations. In plant height growth rate, 17 Agachli accessions were 1.3 times higher on average than that of 10 accessions of Kepele (significant at p = 0.046) and 1.2 times higher for trunk diameter growth rate (p = 0.062). Cluster analysis divided most accessions into two main genetic groups according to their geographic origin. The Agachli group was further divided into two subgroups. One Kepele accession (K4), was genetically different from the rest and clustered on a separate outgroup. Two Agachli accessions (A12 and A17) were outside the two main populations clusters. Accessions K9 and K10 from Kepele were exceptional and were clustered in each of the two Agachli's subgroups, indicating a close genetic relationship between the two populations. In addition, high similarity values (0.58–1.00) and small genetic distances reflect plausible gene flow between Kepele and Agachli, which are 100 Km apart. Mantel test revealed significant relationship between the RAPD and the morphological traits matrices, pointing to the genetic basis for the measured differences in the growth rate. Growing the accessions on the same plot, under similar conditions enabled the evaluation of genotypic differences. The combination of morphological traits and molecular markers will further assist in preservation of genetic variability and cultivation of useful genotypes of P. vera 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.     

Analysis of genetic diversity in Piper nigrum L. using RAPD markers

RAPD analysis was conducted in 22 cultivars of P. nigrum(black pepper) from South India and one accession each of P. longum and P. colubrinum. Twenty-four primers generated 372 RAPD markers of which 367 were polymorphic. Jaccard's similarity between pairs of accessions ranged between 0.11 and 0.66 with a mean of 0.38. Among P. nigrum cultivars, the similarity ranged between 0.20 and 0.66 and the mean was 0.42. The existence of wide genetic diversity as revealed in the present study is supported by earlier reports of extensive inter- and intrapopulation morphological variability in pepper cultivars from South India. UPGMA dendrogram and PCO plot revealed P. colubrinum to be most distant of the three species. Genetic proximity among P. nigrum cultivars could be related to their phenotypic similarities or geographical distribution. Greater divergence was observed among landraces than among advanced cultivars. Landraces grown in southern parts of coastal India and those grown in more northern parts were grouped in separate clusters of the dendrogram.     

Genetic diversity in cucumber (Cucumis sativus L.): I. A reevaluation of the U.S. germplasm collection

Genetic variation within the U.S. cucumber collection (Cucumis sativus var. sativus L. and var. hardwickii (Royle) Alef.) was assessed by examing the variation at 21 polymorphic isozyme loci and comparing the results of this investigation with a similar previous analysis of 14 loci. About 29% (15 of 51) of the enzyme systems examined in an initial survey were polymorphic. Seven loci (Ak-2, Ak-3, Fdp-1, Fdp-2, Mpi-1, Pep-gl and Skdh) which were not previously used to estimate genetic diversity, were assessed. On average, 1.4 loci were polymorphic per enzyme system and 2.2 alleles were present per polymorphic locus. The frequency of polymorphisms was relatively low for Fdp-1(2) (0.01), Mpi-1(1) (0.03), and Skdh(1) (0.02). Principal component and cluster analyses of allelic variation at polymorphic loci separated a diverse array of 757 cucumber accessions from the U.S. National Plant Germplasm Systesm's (NPGS) collection into distinct groups by country (45 nations examined). All accessions of C. s. var. sativus were isozymically distinct from C. s. var. hardwickii, which were themselves dissimilar from each other. Data suggest that C. s. var. hardwickii is not a feral derivative of extant C. s. var. sativus populations. The allelic profile of C. s. var. sativus accessions originating from Burma, Thailand, Indonesia, Hong Kong, Zimbabwe and Ethiopia were distinct from the other accessions examined. Allelic fixation has occurred at Pgd-2 in accessions from Burma, and at Ak-2 in accessions from Zimbabwe and Ethiopia. Some of the countries examined that were in close geographic proximity (e.g., Thailand, Indonesia and Hong Kong) contained accessions with similar isozyme profiles. Accessions are fixed for certain alleles [e.g., Gr(1) (100%), Fdp-1(1) (100%) and Mpi-2(2) (50%) for accessions from Thailand, Indonesia and Hong Kong]. Grouping countries by continent or sub-continent (i.e., North and South American, China, Eastern Europe, Western Europe) and by numbers of accessions examined (i.e., India/Burma, Iran, Japan, Turkey, and remaining accessions) was used to identify accessions with unique allozymic profiles [PIs 209064 (USA), 257486 (China), 188749 (Egypt), 285607 (Poland), 369717 (Yugoslavia), 357844 (Poland), 255936 (Netherlands), 183127 (India), 200818 (Burma), 200815 (Burma), 137836 (Iran), 227013 (Iran), 227235 (Iran), 451976 (Japan), 181752 (Syria), 181874 (Syria), 169383 (Turkey), 171613 (Turkey)].     

Biochemical and cytological analyses in natural populations ofVicia benghalensis L.

Summary 27 accessions ofVicia benghalensis from different geographical origins constitute the pool on which the present study was performed. Genetic variation among the samples was biochemically and cytologically evaluated: seed storage protein profiles and C-banded karyotypes were analysed from single individuals of each accession. SDS-PAGE has shown the possibility to divide the samples into two groups, each characterized by specific protein profiles. The two patterns were indicated as A and B. From individual seed electrophoresis it was ascertained that samples possessing the pattern A showed a low level of individual variation, while those possessing the pattern B were highly polymorphic, thus suggesting differences in the allogamic rate. The cytological analysis demonstrated the presence of two groups of accessions, one being much richer in heterochromatin as evidenced by C-banding (H⁺) than the other (H^-). The analysis of biochemical and karyological data showed a constant association between pattern A and karyotype H⁻ and between pattern B and karyotype H⁺. On the basis of these results it is proposed to considerV. benghalensis as a highly heteromorphic species, in which two groups may be identified.     

Resistance to powdery mildew in populations of barley landraces from Morocco

Forty-eight populations of barley landraces collected from Morocco were screened for resistance to powdery mildew and a number of different resistance genes were detected. Landraces originated from the collection of the International Center for Agricultural Research in the Dry Areas – ICARDA, Aleppo, Syria. Twenty populations of tested landraces (about 42%) showed resistance reactions and 46 single plant lines were selected. Fourteen of these lines were tested in the seedling stage with 17 and another 32 lines with 23 differential isolates of powdery mildew, respectively. The isolates were chosen according to their virulence spectra observed on the Pallas isolines differential set. Five lines originating from five populations of landraces showed resistance to all prevalent in Europe powdery mildew virulence genes. Thirty-five lines (76%) showed resistance reaction type 2. The distribution of reaction type scores indicated that about 81% of all reaction types observed were classified as powdery mildew resistance (scores 0, 1 and 2). In forty-one lines (89%) the presence of unknown genes alone or in combination with a specific one was detected. Four different resistance alleles (Mlat, Mla6, Mla14 and Mla22) were postulated to be present in the tested lines alone or in combination. Among specific resistance alleles the most common was allele Mlat (resistance Atlas). This allele was postulated to be present in twenty-three (50%) tested lines. The use of new identified sources of resistance to powdery mildew in barley breeding is discussed.     

Fruit characterization of Cocos nucifera L. (ARECACEAE) cultivars from the Pacific coast of Costa Rica and the Philippines

Tall coconut cultivars from the Pacific coast of Costa Rica and the Philippines (San Ramón, Tagnanán, and Laguna), were evaluated for fruit characteristics. Most of the introduced cultivars showed extremely large heterogeneity. A cluster analysis, based on the Ward method, classified the palms into four groups with high internal homogeneity. Some of the evaluated coconut palms from the Costa Rican Pacific area had nut characteristics similar to the San Ramon and Tagnanan palm groups but not to the Laguna group. At the association level used (semipartial R ² = 0.10), another group which included the remaining palms sampled from the Costa Rican Pacific coast was constituted.     

Identification and overcoming barriers between Brassica rapa L. em. Metzg. and B. nigra (L.) Koch crosses for the resynthesis of B. juncea (L.) Czern.

Brassica juncea (2n = 36, AABB) is an amphidiploid derived from its diploid progenitor species B. rapa (2n = 20, AA) and B. nigra (2n = 16, BB). Resynthesis of B. juncea by exploiting the wider gene pool of the present day diploids offers a powerful tool for obtaining novel genetic variation. Hybridization between the two species is difficult because of the presence of crossability barriers. Barriers to hybridization were identified using Aniline Blue Fluorescence (ABF) method. Crosses were classified as having pre- or post-fertilization barriers based on whether less than or more than one per cent ovules showed pollen tube entry. Of the 23 crosses studied, seven crosses all with B. rapa as the female parent showed pre-fertilization barriers. Only 0 to 0.3 per cent ovules were fertilized in these crosses. The remaining 16 crosses, four with B. rapa as the female parent and 12 with B. nigra as the female parent, showed post-fertilization barriers. In crosses with pre-fertilization barriers use of bud pollination and stump pollination ensured or increased fertilization. These two simple techniques were used in combination with ovary-ovule culture to ensure recovery of hybrids. In crosses with post-fertilization barriers, ovary-ovule culture alone helped to obtain hybrids. Hybrids were obtained at higher frequency with the use of direct ovule culture compared to ovary-ovule culture. The F₁ plants obtained in vitro were multiplied on MS medium + 2 mg/l Kn + 0.2 mg/l IAA. These F₁ plants were confirmed as true hybrids through morphology, leaf isozymes and cytology. In all 17 hybrids were obtained of which three were amphidiploids.     

Microsatellite DNA Analysis of Wild Hops, Humulus lupulus L.

To study the relationships and genetic diversity among wild hops, Humulus lupulus, we analyzed 133 samples of wild hops collected from Europe, Asia and North America using polymorphism on 11 microsatellite loci. Although only three primers showed bands in Japanese hops, all other samples showed polymorphic bands at most loci. There were no duplicate genotypes among samples of European, Chinese and North American hops, and each individual hop could be distinguished completely. The phylogenetic tree constructed from DA distance with the UPGMA method showed a large cluster comprised of European hops, although Russian hops from the Caucasus and Altai regions were separate from the European cluster. Chinese and North American samples gave distinct clusters suggesting genetic differentiation. This study has indicated that hop microsatellite DNA is differentiated, and is dependent upon the origin in regions of Europe, Asia and North America.     

Isozyme polymorphism and genetic differentiation in natural populations of an endemic tetraploid species <Emphasis Type="Italic">Avena maroccana</Emphasis> in Morocco

A wild tetraploid oat Avena maroccana Gdgr. was collected from the 11 populations in the periphery of Rommani and Casablanca geographic groups of Morocco. Genetic diversity of the species was investigated using six allozyme systems. Allelic frequencies were scored representing eight polymorphic and five monomorphic loci. Coefficient of gene differentiation (Gst) was 0.3019, which indicated great genetic differentiation. The number of alleles per locus was 2.6154, the percentage of polymorphic loci was 61.54, and the expected heterozygosity was 0.2462 in all populations. Genetic diversity in A. maroccana was high in comparison to self-pollinated species. In total, nine heterozygotes resulting from outcrossing were found in the progeny from M1, M3, M4, M22 and M26. The population of M7 had peculiar alleles Pgd–2SS and Pgd-1SS in high frequency. M9 had the lowest level of diversity out of the 11 populations. Geographic and genetic distances between all the populations were not significantly correlated with each other (r = 0.0996). Cluster analysis showed that two groups, (M1, M22, M2 and M4) and (M3, M23, M8, M5 and M26) were apparently differentiated. Two populations of the Casablanca group, M7 and M9 were independent from each other, and were separated distinctly from the other populations. Genetic diversity of the Rommani and Casablanca groups was almost the same in all the parameters. This was due to the similar man-made habitat such as roadside or rich fertile soil and brown clay soils. The population size of A. maroccana was small and restricted to the narrow central Morocco with great genetic differentiation so that genetic diversity may be reflected from the results of genetic drift and outcrossing heterozygote segregation.