Cytogenetic study of F1 hybrids betweenCrepis dinarica andCrepis froelichiana (Asteraceae)

Crepis dinarica andC. froelichiana are two closely related species of theC. praemorsa complex. Even though they exhibit the same chromosome number (2n = 8) and similar idiogram shape, they differ widely in quantity and distribution of heterochromatin bands. The hybrids between these two species comprise three morphological types. Parental genomes were distinguished in hybrids by Giemsa differential staining (C-banding). Although meiosis presents only a few abnormalities (about 2.4%), the percentage of aborted pollen grains is very high (90%).     

玉米8个栽培亚种（类型）的核型和C—带带型的比较研究

本文首次报道了玉米8个亚种、2个亚型和2个杂交品种的核型和Giemsa C-带带型。所有材料的根尖细胞染色体数目均为2n=20。主要由中部和亚中部着丝点染色体组成。第6染色体短臂均具随体,但大小不同。所有材料均显示有亚端带和端带,在第6染色体的短臂上显示有NOR或/和随体带。C-带的分布、总数目和总长度各不相同。其总带数变异于6至18之间,C-带总长度为5.65—11.40％之间。在核型中,具中部着丝点的染色体数目及C-带总数,罕见栽培或原始的类型通常多于广泛栽培的类型。此外,有关核型和C-帝的变异和进化也进行了简略的讨论。     

Molecular genetic diversity of the Turkish national hazelnut collection and selection of a core set

European hazelnut (Corylus avellana L.) is an economically and nutritionally important nut crop with wild and cultivated populations found throughout Europe and in parts of Asia. This study examined the molecular genetic diversity and population structure of 402 genotypes including 143 wild individuals, 239 landraces, and 20 cultivars from the Turkish national hazelnut collection using simple sequence repeat (SSR) markers. A total of 30 SSR markers yielded 407 polymorphic fragments. Diversity analysis of the Turkish hazelnut genotypes indicated that they fell into three subpopulations according to ad hoc statistics and neighbor-joining algorithm. Although all cultivars clustered together, they overlapped with the wild accessions and landraces. Thus, the dendrogram, principal coordinate, and population structure analyses suggest that they share the same gene pool. A total of 78 accessions were selected as a core set to encompass the molecular genetic and morphological diversity present in the national collection. This core set should have priority in preservation efforts and in trait characterization.     

Inter- and intraplant C-banding polymorphism in one population ofAegilops comosa subsp.comosa var.comosa (Poaceae)

The Giemsa C-banding pattern of the chromosomes of the native self-pollinatedAegilops comosa subsp.comosa var.comosa was studied. Six of the seven chromosomes of the haploid genome were found to be polymorphic for C-banding patterns. Chromosome A had four variants, chromosome E three variants and each of the chromosomes B, D, and F two variants. Chromosomes E and G were polymorphic for arm length and arm ratio.This paper is part of the doctoral dissertation ofA. Georgiou.     

Genetic Variability in Pepper (<Emphasis Type="Italic">Capsicum annuum</Emphasis> L.) Estimated by Morphological Data and Amplified Fragment Length Polymorphism Markers

Data on genetic similarity among crop cultivars is of vital importance for the plant breeder. The objectives of this study were to group pepper (Capsicum annuum L.) genotypes into clusters according to their distances as estimated by morphological traits and amplified fragment length polymorphism (AFLP) markers and to assess the relationships between the two. Thirty-nine pepper genotypes obtained from different countries were grown in the greenhouse at University of the Free State, South Africa, during 2001 and 2002 in a randomized complete block design with three replications. A total of 20 different morphological traits were measured and six AFLP primer pairs were used to estimate pairwise genetic distances. Both datasets showed high genetic distances among the different genotypes, indicating high genetic diversity among them. The mean genetic distance among Ethiopian pungent elongated-fruit genotypes, was lower than that between them and the introduced ones. Morphological and AFLP distance estimations generally clustered together genotypes with similar fruit sizes. Significant, positive correlation was observed between morphological and AFLP diversity estimations. The narrow genetic basis among the Ethiopian pungent elongated-fruit cultivars suggests that the pepper breeding program of Ethiopia should focus on enriching its germplasm through local collection and introductions from other parts of the world.     

Characterization of ten date palm (Phoenix dactylifera L.) cultivars from Saudi Arabia using AFLP and ISSR markers

Date palm is the most economically important plant in the Middle East due to its nutritionally valuable fruit. The development of accurate DNA fingerprints to characterize cultivars and the detection of genetic diversity are of great value for breeding programs. The present study explores the usefulness of ISSR and AFLP molecular markers to detect relationships among 10 date palm (Phoenix dactylifera L.) cultivars from Saudi Arabia. Thirteen ISSR primers and six AFLP primer combinations were examined. The level of polymorphism among cultivars for ISSRs ranged from 20% to 100% with an average of 85%. Polymorphism levels for AFLPs ranged from 63% to 84% with an average of 76%. The total number of cultivar-specific markers was 241, 208 of which were generated from AFLP analysis. AJWA cultivar had the highest number of cultivar-specific ISSR markers, whereas DEK, PER, SUK-Q, SHA and MOS-H cultivars had the lowest. RAB and SHA cultivars had the most and least AFLP cultivar-specific markers, respectively. The highest pairwise similarity indices for ISSRs, AFLPs and combined markers were 84% between DEK (female) and PER (female), 81% between SUK-Q (male) and RAB (male), and 80% between SUK-Q (male) and RAB (male), respectively. The lowest similarity indices were 65% between TAB (female) and SUK-Q (male), 67% between SUK-A (female) and SUK-Q (male), and 67% between SUK-A (female) and SUK-Q (male). Cultivars of the same sex had higher pairwise similarities than those between cultivars of different sex. The Neighbor-Joining (NJ) tree generated from the ISSR dataset was not well resolved and bootstrap support for resolved nodes in the tree was low. AFLP and combined data generated completely resolved trees with high levels of bootstrap support. In conclusion, AFLP and ISSR approaches enabled discrimination among 10 date palm cultivars of from Saudi Arabia, which will provide valuable information for future improvement of this important crop.     

Geographical trends within a diverse spring barley collection as identified by agro/morphological and electrophoretic data

For ex-situ germplasm conservation purposes, the concept of genetic diversity being concentrated in certain geographic regions is useful for the conscious selection of diverse forms. Numerous studies of barley and other major corp species often confirm the concentration of simply-inherited, phenotypicallyobvious markers within the Vavilovian centres of diversity/origin. However, more recent studies of electrophoretic patterns and or more complexly-inherited traits do not always confirm the same geographic patterns. Unfortunately, few studies of world germplasm collections have screened a range of agro/morphological/electrophoretic patterns using the same germplasm collection as a consistent base for evaluation purposes, making precise genetic estimates of diverse geographic areas difficult. A diverse collection of 1 118 spring-sown barley cultivars was, therefore, evaluated for both agro/morphological and biochemical genetic markers in an effort to identify appropriate criteria for the construction of a comprehensive ex-situ germplasm collection. On the basis of both agro/morphological and biochemical data, countries whose cultivated barley germplasm was identified as diverse and genetically distinct were Algeria, Afghanistan, Argentina, Ethiopia, India, Peru and Turkey. However, within broad limits, separate cluster analyses of the agro-morphological and electrophoretic patterns identified dissimilar groups of countries, which demonstrated that a collection strategy based solely on country of origin is inappropriate.     

Comparisons between C-banding Patterns on Root Tip Chromosomes of Different Cultivars in Maize( Zea mays)

The technique of Giemsa banding, C-banding patterns on the root tip chromosomes and the chromocentres of interphase nuclei with three eultivars (Dai Zi Bai, Qun Dan 105 and 2×6) of maize were studied. The results are as follows: 1. After fixation and treatment in a saturated solution of barium hydroxide the preparations were incubated in 0.5×SSC, I×SSC, 3×SSC, 4×SSC or distilled water respectively forⅠ h at 60 ℃ and the other steps in C-banding procedure were not changed so as to find the optimum saline concentration for Giemsa banding in maize. The experimental results shown that 0.5×SSC was the best. But bands could not produced very well by treating samples in distilled water. 2. There were terminal, subterminal and centric bands in Dai Zi Bai and Qun Dan 105. The C-banding patterns on the root tip chromosomes of these two cultivars were different from each other. Qun Dan 105 had 10 prominent bands in total, while Daf Zi Bai had 7. The banding patterns of each chromosome were described in detail. 3. The average chromocentres per cell in Dai Zi, Qun Dan 105 and cultivar 2×6 were 7.1, 10.9 and 7.2 respectively. Their prominent band numbers on the chromosomes were 7, 10 and 8 correspondingly. It seems that the number of C-bands on the chromosomes is close to that of chromocentres.     

基于AFLP分子标记的桂花品种核心种质的构建

用100个桂花品种荧光AFLP分子标记信息构建核心种质。利用获得的指纹信息,运用UPGMA聚类取样法,采用Kimura 2-parameter遗传距离,多次聚类随机抽样。结果表明:(1)8对引物共获得514条带,平均每对引物获得64条带。(2)从100个桂花品种中筛选了30个样本的核心种质。(3)比较核心种质和全部种质的Shan-non-Wiener指数(H′)和Simpson指数(D),t检验值均说明核心种质的遗传多样性指数与全部种质遗传多样性没有明显差异,表明所构建的核心样品能够很好地保留原始100个桂花品种的遗传多样性。     

Morphological traits and RAPD markers for characterization and identification of minor Spanish olive cultivars from the Extremadura region

A total of 91 wild olive accessions and 31 olive cultivars growing in the Extremadura region of central-western Spain were analyzed using morphological traits and RAPD markers. We focused on three main and 16 minor Spanish olive cultivars that are recognized as native or local to the Extremadura region. The five arbitrary 10-mer primers tested on the olive cultivars gave 67 polymorphic bands, representing 91% of the total amplification products. The number of bands per primer ranged from 9 to 18, whereas the number of polymorphic bands ranged from 8 to 17. All the cultivars could be identified by a combination of three primers (OPF-6, OPA-8, and OPK-16); four cultivar-specific markers were detected. The minor local "Jariego" and "Tempranillo" cultivars showed the most distal similarities. The resulting dendrogram, using the unweighted pair-group method with arithmetic mean clustering algorithm, depicted the pattern of relationships between the local Extremadura cultivars and the cultivars from geographically connected regions. This analysis showed a correlation between most of the minor local cultivars and the geographical origin; there was no apparent clustering according to morphological traits or fruit use of olive cultivars when these parameters were used as analysis criteria.     

Congruence between morphological and molecular markers inferred from the analysis of the intra-morphotype genetic diversity and the spatial structure of Oxalis tuberosa Mol.

Oxalis tuberosa is an important crop cultivated in the highest Andean zones. A germplasm collection is maintained ex situ by CIP, which has developed a morphological markers system to classify the accessions into morphotypes, i.e. groups of morphologically identical accessions. However, their genetic uniformity is currently unknown. The ISSR technique was used in two experiments to determine the relationships between both morphological and molecular markers systems. The intra-morphotype genetic diversity, the spatial structures of the diversity and the congruence between both markers systems were determined. In the first experience, 44 accessions representing five morphotypes, clearly distinct from each other, were analyzed. At the molecular level, the accessions exactly clustered according to their morphotypes. However, a genetic variability was observed inside each morphotype. In the second experiment, 34 accessions gradually differing from each other on morphological base were analyzed. The morphological clustering showed no geographical structure. On the opposite, the molecular analysis showed that the genetic structure was slightly related to the collection site. The correlation between both markers systems was weak but significant. The lack of perfect congruence between morphological and molecular data suggests that the morphological system may be useful for the morphotypes management but is not appropriate to study the genetic structure of the oca. The spatial structure of the genetic diversity can be related to the evolution of the species and the discordance between the morphological and molecular structures may result from similar selection pressures at different places leading to similar forms with a different genetic background.     

Heterochromatin and silver banding of rye (Secale cereale,Gramineae) chromosomes

Air-dried chromosomes of rye when stained with aqueous silver nitrate show differential banding patterns. In addition to staining the NOR sites, the silver nitrate stains all regions of constitutive heterochromatin, as identified by Giemsa C-banding, as well as a number of small interstitial regions. However, the heterochromatin on the B chromosome is not stained by the silver method. This is proposed as a rapid and reliable banding method.     

Comparison of rapeseed cultivars and resynthesized lines based on allozyme and RFLP markers

It has frequently been suggested to use the resynthesis of rapeseed (Brassica napus) from B. campestris and B. oleracea to broaden its genetic base. The objective of the present study is twofold: (1) to compare the genetic variation within resynthesized rapeseed with a world-wide collection of oilseed rape cultivars, and (2) to compare genetic distances estimated from RFLP markers with distances estimated from a relatively small number of allozyme markers. We investigated 17 resynthesized lines and 24 rapeseed cultivars. Genetic distances were estimated either based on the electrophoresis of seven allozymes, with a total of 38 different bands, or based on RFLP data of 51 probe/enzyme combinations, with a total of 355 different bands. The results of allozyme and RFLP analyses agreed reasonably well. Genetic distances, estimated from two independent sets of RFLP data with 25 and 26 probe/enzyme combinations respectively, were highly correlated; hence about 50 RFLP markers are sufficient to characterize rapeseed material with a large genetic diversity. The cultivars were clustered into three groups: (1) spring rapeseed of European and Northern American origin, (2) winter rapeseed of European and Northern American origin, and (3) rapeseed of Asian origin. Several of the resynthesized rapeseed lines were similar to European winter rapeseed cultivars, whereas others had quite unique patterns. It is concluded, that resynthesized rapeseed is a valuable source for broadening the genetic variation in present breeding material of Brassica napus. However, different lines differ widely in their suitability for this purpose.     

Chromosome banding and genome size differentiation inProspero (Hyacinthaceae): Diploids

Prospero is a Mediterranean autumn-flowering genus ofHyacinthaceae commonly classified inScilla asS. autumnalis andS. obtusifolia. Extensive dysploid and polyploid variation has been reported. In the present study 77 diploid accessions from the western to the eastern part of the area of distribution, the major part being from continental Greece and Crete, have been analysed for karyotype structure and, in part, for genome size. Methods employed were acetocarmine staining, Giemsa C-banding, fluorochrome staining mainly with chromomycin A₃/DAPI, silver impregnation, and Feulgen densitometry. Banded idiograms were established with a computer assisted karyotype analysis procedure. Chromosome numbers were 2n = 8 inP. obtusifolium, and 2n = 12 and 14 inP. autumnale s. l. Dispensable euchromatic chromosome segments and different types of B chromosomes occurred. Among the cytotypes with 2n = 14 two karyotypes from Turkey differed from each other and from the rest in form, position of the nucleolar constriction, and in genome size. The remaining accessions were similar in karyotype shape but three levels of genome size could be discerned, the highest (1C = 7.50 pg) being found on the Iberian Peninsula, an intermediate one on Corsica and Malta, and the lowest (4.27 pg) in the Aegean. The karyotype with 2n = 12 had an intermediate genome size, and that ofP. obtusifolium a relatively low one. Heterochromatin amount was generally low, but some karyotypes showed characteristic banding patterns. The relationship between the chromosome complements with 2n = 14, 12 and 8 is discussed on the basis of idiograms and DNA amounts.The authors respectfully dedicate this papers to emer. o. Prof. Dr.Elisabeth Tschermak-Woess on the occasion of her 80th birthday.     

Detection of interspecific and intraspecific variation in Panicum millets through random amplified polymorphic DNA

The potential use of random amplified polymorphic DNA (RAPD) was evaluated as a source of genetic markers for studying variation among four species of Panicum and within the crop species P. miliaceum and P. sumatrense. Polymorphism in RAPD markers was observed across and within species. The four species were distinct in RAPD patterns and were separated at low correlation values even with small samples involving single genotypes per species. Accessions of P. miliaceum were grouped according to geographical regions of origin. The study demonstrated that unlike isozyme and protein electrophoresis patterns, RAPD markers can be applied to studying genetic diversity, defining gene pools, and identifying cultivars for this group of millets.     

Morphological and allozyme variation in a collection of Cucumeropsis mannii Naudin (Cucurbitaceae) from Côte d'Ivoire

To set up a rational collecting strategy for germplasm of the edible-seeded cucurbit Cucumeropsis mannii, a study was conducted using 24 morphological and seven putative enzyme markers to determine the intra-specific variability from 16 and 22 accessions (representing three cultivars), respectively. The analysis of variance, showed a significant difference between the three cultivars. Principal component analysis pointed out a variation among individuals, mainly on the basis of flower, fruit, and seed size. Dendrogram with UPGMA method allowed clustering of the cultivars. Genetic diversity indices estimated equalled: 9.96% for the proportion of polymorphic loci (P), 1.10 for the number of alleles (A) and 0.023 for observed heterozygosity (H_o). The level of the within accessions genetic diversity (H_S = 0.078) was higher than among accessions (D_ST = 0.042). Nei's genetic distances between the three cultivars were also low (0.079–0.147), indicating a high degree of similarity of the analysed cultivars.     

Comparative analysis of C-band karyotypes inCrepis praemorsa subsp.praemorsa and subsp.dinarica

Crepis incarnata subsp.dinarica (G. Beck)Hayek is nowadays considered as a subspecies ofC. praemorsa (L.)Tausch. Comparative analyses of Feulgen karyotypes demonstrate great similarities, but remarkable differences in the presence and the distribution of the constitutive heterochromatin in the two taxa are detected by using the Giemsa differential staining technique (C-banding). This favours their specific distinctness.     

Cytogenetical characterization of Odontesthes bonariensis (Pisces,Atherinidae), an Argentine species introduced in Italy

Giemsa and Ag-staining techniques and C-banding were performed on cytological preparations of 22 specimens of silversides (Odontesthes bonariensis) in order to extend the karyological knowledge on the family Atherinidae, to contribute cytotaxonomically to the systematics of the species and to identify possible cytogenetical markers of the introduced population.The karyotype is conservative, 2n being 48, and 44 chromosomes being uniarmed. The remaining four biarmed chromosomes show intraindividual morphological variability. This variability is sex-independent. Ag-stanining techniques reveal that NORs are located on the short arms of biarmed chromosomes and that the morphological variability is partially due to NOR activity. C-banding revealed the associations between NORs and constitutive heterochromatin which could be responsible for some apparent structural differences. The differences found could be cytogenetical markers of the introduced population.Research work supported by CNR, Italy. Special grant I.P.R.A. — Sub-project 1. Paper No. 1192.