Genotypic variation in field resistance of Cucurbita pepo cultivars to powdery mildew (Erysiphe cichoracearum)

Summary Fifty-eight cultivars of Cucurbita pepo (mostly zucchini type) were studied in three-year experiments for field resistance to powdery mildew caused by Erysiphe cichoracearum. No genotypes were found with complete resistance to powdery mildew. There were significant differences in the expression of field resistance characterized by some basic epidemiological parameters (maximum infection degree, area below curve). The highest level of field resistance occurred in 13 cultivars including Acceste F1, Albina, ambassador F1, CU-235, Elite F1, Goldfinger, Parmanta F1, Seneca Hybr. F1.Abbreviations ABC area below curve - ID infection degree - CO Cocozelle - VM Vegetable marrow - ZU Zucchini     

Seed Dimensions in the Subspecies and Cultivar-groups of Cucurbita pepo

Seed samples of 174 accessions of pumpkins, squash, and gourds of Cucurbita pepo were measured for length, width, and thickness. Mean seed length ranged from 8.8 to 23.3 mm, mean seed width from 5.0 to 12.5 mm and mean seed thickness from 1.2 to 3.8 mm. The seeds of the edible-fruited cultivar-groups of C. pepo subsp. pepo (Pumpkin, Vegetable Marrow, Cocozelle, Zucchini) were longer, wider, and thicker than those of their counterparts of C. pepo subsp. texana (Acorn, Crookneck, Scallop, Straightneck). The small-fruited, inedible gourds had the smallest seeds. Of the edible-fruited C. pepo subsp. pepo, the Pumpkin Group (round fruits) possessed the longest and flattest seeds whilst the Zucchini Group (long, uniformly cylindrical fruits) possessed the shortest and thickest seeds. The samples tested also differed greatly in mean values for relative dimensions. Length-to-width ratio ranged from 1.5 to 2.5, length-to-thickness ratio from 3.7 to 8.2, and width-to-thickness ratio from 2.0 to 4.9. The Pumpkin Group, followed by the Cocozelle Group (long, bulbous cylindrical fruits) and the Vegetable Marrow Group (short, tapered cylindrical fruits), had seeds of the highest length-to-width ratio. The seeds of the four edible-fruited groups of C. pepo subsp. pepo differed significantly from one another in their length-to-thickness and width-to-thickness ratios. The seeds of the Pumpkin Group and the wild Mexican gourd (C. pepo subsp. fraterna) differed greatly in their dimensions but were most similar to one another in their relative dimensions. In C. pepo, seed size is positively associated with fruit size and seed shape is inversely associated with fruit shape.     

AFLP analysis of genetic diversity in five accessions of Polish runner bean (<Emphasis Type="Italic">Phaseolus coccineus</Emphasis> L.)

Runner bean (Phaseolus coccineus L.) is traditionally cultivated in Poland for dry seeds. The national collection of runner bean maintained in the National Center for Plant Genetic Resources gathers 152 accessions, which are mainly landraces originated in Poland (68%), Ukraine (17%) and Slovakia (10%). The collection contains valuable genetic resources for bean breeders and research. The aim of this study was to describe the level and structure of genetic diversity of three landraces and two commercial cultivars of runner bean from the national collection in order to assess their genetic potential for breeding. Amplified Fragment Length Polymorphism analysis included five combinations of selective primers. Analysis of seven genetic diversity parameters reveled fair amount of genetic variation both in landraces and cultivars. High genetic diversity of commercial cultivars relative those of landraces suggests that the breeding process leading to their release was rather moderate and most likely included domestic gene pool of runner bean. Low gene diversity and low Nei’s genetic distance values as well as intergradations among accessions in the PCoA may indicate reduced variability P. coccineus grown in Poland as a result of its migration pathways.     

Population genetics of traditional landraces of <Emphasis Type="Italic">Cucurbita pepo</Emphasis> L., 1753 in the cloud forest in Baja Verapaz,Guatemala

Little is known regarding the effect of fragmentation and human agricultural management on the genetic variation and gene flow of Cucurbita pepo L., 1753 in moderate fragmented areas in central Guatemala. We hypothesize that the genetic variation of C. pepo is affected by forest fragmentation and by traditional agricultural management. Therefore, we aim to determine: (1) the genetic diversity and genetic structure of C. pepo in the Cloud Forest Corridor (CFC) (2) the extent of genetic admixture between commercial variety (CV) and traditional landraces (TL) of C. pepo, (3) the effect of habitat fragmentation in the population genetics of C. pepo with a landscape approach, and (4) the potential relationship between traditional management practices and genetic diversity of C. pepo in the CFC. We detected the existence of high level of genetic diversity (AR = 3.43; He = 0.50), inbreeding (Fis = 0.25) and moderate population structure of C. pepo in the CFC (Fst = 0.16). No correlation between landscape and genetics was found. Also, we found high genetic admixture between CV and TL. It seems that human practices, mainly related with seed exchange patterns, could affect genetic diversity of C. pepo in the CFC. C. pepo populations in the CFC are structured, with inbreeding, and show admixture with the CV, an aspect that could affect its genetic diversity. The agricultural management influenced the population genetics of C. pepo in the CFC, but the landscape did not. We suggest that special efforts should be made to preserve the diversity of this important indigenous food source for Guatemalan people as well as their management practices.     

Variation in melon (<Emphasis Type="Italic">Cucumis melo</Emphasis>) landraces adapted to the humid tropics of southern India

We present here the first comprehensive genetic characterization of melon landraces from the humid tropics of southern India. The genetic diversity among 50 melon landraces collected from 3 agro-ecological regions of southern India (6 agro-ecological sub-regions) was assessed by variation at 17 SSR loci, morphological traits of plant habit and fruit, 2 yield-associated traits, pest and disease resistance, biochemical composition (ascorbic acid, carotenoids, titrable acidity) and mineral content (P, K, Fe, Zn). Differences among accessions were observed in plant and fruit traits. Melon germplasm with high titrable acidity, higher than average amounts of mineral content and resistance to Cucumber mosaic virus, Zucchini yellow mosaic virus, powdery mildew (races 1, 2, 3, 5), Fusarium wilt (races 1, 2), Aphis gossypii and leafminer was recorded in the collection. A high level of genetic variability in melon germplasm was suggested by the SSR analysis. Comparative analysis using SSRs of the genetic variability between Indian melons from north, south, and east regions and reference accessions of melon from Spain, France, Japan, Korea, Iraq, Zambia showed regional differentiation between Indian melon accessions and that Indian germplasm was weakly related to the melon accessions from other parts of the world, suggesting that an important portion of the genetic variability found within this melon collection has not been used yet for the development of new cultivars. Additional collections of acidulus melon germplasm should be made in southern India and adequate management of this important genetic resource is clearly a necessity.     

Spanish melons (Cucumis melo L.) of the Madrid provenance: a unique germplasm reservoir

Melon (Cucumis melo L.) landraces of the Madrid provenance, Spain, have received national distinction for their high fruit quality and sensorial attributes. More specifically, a unique array of Group Inodorus landraces have been continuously cultivated and conserved by farmers in the municipality of Villaconejos since the 19th century. Their genetic relationships to other Group Inodorus and Flexuous melon market classes is not known, and, thus, a study was designed to determine their genetic relationships using 52 simple sequence repeat (SSR) markers, and then make genetic comparisons between these accessions and a previously published “Standard Reference Germplasm Array” (RA) containing Group Inodorus (14 Spanish and one USA), Flexuosus (1 Spanish), and Cantalupensis ( 2 USA) melon accessions. This subset consisted of 15 Spanish Group Inodorus landraces that circumscribed the genetic variation of major Spanish melon market classes (Groups Inodorus and Flexuosus), and USA commercial varieties (Groups Cantalupensis and Inodorus). Based on genetic distances, Villaconejos (Madrid) genotypes differed substantially from RA subset accessions, thus defining their genetic uniqueness. Principal component analysis (PCA) partitioned the accessions examined into four distinct groups revealing that Villaconejos black epidermis melons (landraces ‘Largo’, ‘Largo Negro Escrito’ and ‘Puchero’) were distinctly different from all other accessions examined, as cluster analysis separated Rochet market type Villaconejos’ accessions (landraces ‘Mochuelo’, ‘Mochuelo Tradicional’ and ‘Melón de Villaconejos’) from RA of the same market type. Genetic assessment of principal Spanish market classes revealed comparatively low intra-market heterogeneity in Piel de Sapo type accessions and high heterogeneity in Black and Yellow market type accessions. While a relatively high level of genetic introgression was detected between Yellow and Green market types, black epidermis market types were genetically unique. Given the uniqueness and high genetic diversity resident in Villaconejos landraces, this germplasm pool should be considered as a genetic source for broadening the comparatively narrow genetic base of Group Cantalupensis and Inodorus melon market types, especially standard commercial Spanish Group Inodorus market types (e.g., Piel de Sapo, Rochet, and Canari).     

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.     

Genetic variation in common and runner bean of the Northern Meseta in Spain

The genetic variation within and between Spanish landraces or varieties of Phaseolus vulgaris L. (common bean) and P. coccineus L. (runner bean) has been estimated by means of isozymes and random amplified polymorphic DNA (RAPD) analyses. Likewise, storage protein and amino acid content in dry seeds have been estimated. Fifteen landraces (60 accessions) of P. vulgaris and six of P. coccineus (six accessions) have been studied. Of the seven isozymatic systems analyzed only three systems and three loci showed variability in each species. Isozyme analyses revealed that genetic variability within and between landraces exist in both species. Even variability within accession was detected in some P. vulgaris landraces. Comparison of isozyme data indicated that Spanish landraces have a lower level of genetic variability than wild American materials and probably also lower than American landraces. RAPD analysis allowed for the uniquely distinguishing of all landraces. Genetic similarity among landraces, estimated by both isozymes and RAPDs, were not related with the seed morphological characters (color, size and shape) which define each variety or landrace. Variation in protein and amino acid content among landraces was also detected. The average protein content in common bean (20.48%) was similar to values previously reported in this species and higher than the average in the runner bean landraces (16.33%). In relation to the amino acid content methionine and cysteine showed the lowest values in all samples, although the content of these two amino acids varied widely among landraces.     

Genetic, morphological and cyanogen content evaluation of a new collection of Caribbean Lima bean (Phaseolus lunatus L.) landraces

Lima bean is a species cultivated broadly in the Americas and has been cultivated in the Caribbean for at least 500 years. In order to determine the genetic structure and diversity of Lima bean from the Caribbean, 50 landraces from Haiti, the Dominican Republic and Puerto Rico were collected and analyzed using 24 SSR markers. All landraces in this Caribbean collection were found to be of Middle-American descent. The genetic diversity (H_E) was highest in the landraces from Puerto Rico and lowest in Haitian landraces. The observed proportion of heterozygotes (H_O) was higher in the Haitian landraces and lowest in Puerto Rican landraces. Unweighted pair-group method with arithmetic averaging analysis showed that the landraces clustered into 3 clusters with all Haitian landraces grouping in one cluster. We also assessed the agro-morphological characteristics of the collection as well as the content of cyanogenic glucoside, linamarin, in leaves and dry seeds. Lima bean, which is a model crop for indirect plant defenses against herbivory, also possess linamarin as a source of direct plant defense. Upon tissue damage, linamarin is converted to toxic hydrogen cyanide. In our collection 44.6 % of the landraces had average seed HCN content ≤200 ppm which is the permitted level for Lima bean seed in the US. Our results also identified the landraces in this collection which have high linamarin levels in the leaves while having low levels in the seeds. Such landraces have the desirable combination of traits and will be the focus of our future plans for agronomic trait improvement though breeding.     

The Sierra Norte of Madrid: an agrobiodiversity refuge for common bean landraces

At the mountain area close to the city of Madrid, common beans were one of the main food crops present in everyday diet until 1960. This paper describes the morphological diversity, for forty-six phenological and morphological traits, found in forty-three traditional varieties of common beans collected in this area and seven commercial varieties used as reference. That comparison suggests that common bean breeding programs have led to later varieties with a higher production of straight pods. Quantitative traits showed also similar values when compared to the Iberian common bean collection, except for the phenological and seed size characters. The phenological differences could suggest an ecological adaptation of the studied landraces to the regional environmental conditions. The different seed size averages of both collections might correspond to the scarce presence of smaller seed-type common beans in Madrid collection. All the accessions collected in the Sierra Norte of Madrid belong to any of the groups included in the Spanish core collection. Madrilenian collection is also composed by indeterminate growth habit varieties, while the presence of bush accessions is relatively uncommon. Most of the Madrilenian landraces have a remarkable fitness for green-pod consumption (42 %) and their seeds are mainly white (30 %), ovate-shape (49 %) and medium-large (40 %). The study of seed storage protein allowed to classify the landraces according to their domesticated gene pools. Most of them (72 %), with T and C phaseolin type, seem to belong to Andean germplasm, while the remaining (28 %), with S and B phaseolin type, to Mesoamerican one. The remarkable morphological diversity of common beans found in this small area is a symptom of a broad genetic base despite genetic erosion, what indicates a widespread crop in the past. Therefore, it is advisable to design agro-environmental policies to promote the production and commercialization of common bean landraces in Sierra Norte of Madrid.     

Italian common bean landraces: diversity and population structure

Phaseolus vulgaris L. is an important species that originated in Mesoamerica. A Mesoamerican and an Andean gene pool are usually distinguished in the domesticated forms. Many bean landraces are still cultivated in Italy and the Department of Applied Biology maintains an ex situ collection of 146 landraces. Although protection schemes are being developed in Italy, most landraces are extinct or at risk of extinction. To facilitate their conservation and use, geographical, morphological, biochemical and molecular (SSR) data were collected and analysed to estimate the diversity and the genetic structure of the collection. Data confirmed that both the Mesoamerican and the Andean gene pools were introduced in Italy and, although a distinction between the two gene pools exists, the Italian landrace diversity is clearly structured in three clusters that are not simply ascribable to the original gene pools. The observed structure appears also to be due to adaptation to the different environmental conditions determined by altitude. This was confirmed by assessing the presence of selective effects for some of the SSR used in this study. Finally, a certain extent of admixture in Italian landrace diversity suggests past (or recurring) hybridisation events among gene pools. The combined use of morphological, biochemical and molecular data clearly distinguished almost all the landraces. The data gathered here can assist landrace in situ protection schemes that are being developed in Italy, be used to register landraces in the European common catalogue of ‘conservation varieties’ for seed commercialisation and contribute to a better use of Italian common bean diversity in breeding for organic and conventional production systems.     

Genetic diversity analysis of yams (Dioscorea spp.) cultivated in China using ISSR and SRAP markers

Yam (Dioscorea spp.) is widely cultivated in China and many landraces are maintained by local farmers. However, there is little information available about their diversity and species identity. In this study, inter simple sequence repeat (ISSR) and sequence related amplified polymorphism (SRAP) techniques were used to assess genetic diversity within 21 yam landraces from seven cultivated populations. We observed high level of polymorphism among these landraces, specifically, 95.3 % for ISSR and 93.5 % for SRAP. Analysis of molecular variance revealed a significantly greater variation among the four yam species (40.39 %) and their populations (35.78 %) than within the populations (23.83 %). The unweighted pair group method arithmetic averages clusters and principal component analysis for 21 landraces formed four well-separated groups containing landraces of each of the four species, namely, Dioscorea opposita Thunb., Dioscorea alata L., Dioscorea persimilis Prain et Burkill, and Dioscorea fordii Prain et Burkill. The ISSR and SRAP primers were highly discriminatory among the 21 landraces; all 21 landraces could be easily differentiated using these primers. The average mean of gene flow (Nm = 0.1081) estimated from high genetic differentiation (Gst = 0.8222) suggested that gene flow among the populations was relatively restricted. The lack of genetic diversity within individual yam species suggests that it is critical to develop long-term strategies for enhancing genetic diversity within various yam species.     

Population structure and genetic differentiation among the USDA common bean (Phaseolus vulgaris L.) core collection

Genetic diversity data were collected from a large population of common bean (Phaseolus vulgaris L.) landraces representing the United States Department of Agriculture core collection. The data were based on microsatellite data from all linkage groups. A procedure was developed to determine if we collected sufficient marker data to adequately estimated pairwise diversity. The diversity data were used to define populations using distance and model-based approaches. Genetic differentiation and genetic isolation by distance data were collected. Diversity was also compared for markers linked and unlinked to domestication loci. Using a model-based approach, the landraces were divided into the traditional Middle American and Andean gene pools. Diversity was greater for the Middle American gene pool. Six Middle American and three Andean subpopulations were defined, and the Middle American subpopulations exhibited strong geographic identity. Unlike other studies, seed size varied considerably with subpopulations, and a number of the subpopulations contained landraces from multiple common bean races. All of the subpopulations were highly differentiated, with the Middle American subpopulations showing the greatest differentiation. Genetic isolation by distance was observed among the Middle American and Andean subpopulations but not among subpopulations within a gene pool. Within each gene pool, diversity was lower for markers linked to domestication loci.     

Combined use of gliadins and SSRs to analyse the genetic variability of the Spanish collection of cultivated diploid wheat (Triticum monococcum L. ssp. monococcum)

This work studied the combined use of gliadins and SSRs to analyse inter- and intra-accession variability of the Spanish collection of cultivated einkorn (Triticum monococcum L. ssp. monococcum) maintained at the CRF-INIA. In general, gliadin loci presented higher discrimination power than SSRs, reflecting the high variability of the gliadins. The loci on chromosome 6A were the most polymorphic with similar PIC values for both marker systems, showing that these markers are very useful for genetic variability studies in wheat. The gliadin results indicated that the Spanish einkorn collection possessed high genetic diversity, being the differentiation large between varieties and small within them. Some associations between gliadin alleles and geographical and agro-morphological data were found. Agro-morphological relations were also observed in the clusters of the SSRs dendrogram. A high concordance was found between gliadins and SSRs for genotype identification. In addition, both systems provide complementary information to resolve the different cases of intra-accession variability not detected at the agro-morphological level, and to identify separately all the genotypes analysed. The combined use of both genetic markers is an excellent tool for genetic resource evaluation in addition to agro-morphological evaluation.     

Agro-morphological diversity of Spanish traditional melons (<Emphasis Type="Italic">Cucumis melo</Emphasis> L.) of the Madrid provenance

The variability of 14 landraces belonging to a Madrilean village historically specialized on melon cultivation, Villaconejos, was evaluated based on 58 quantitative and qualitative morphological traits. These landraces were compared to a reference array composed of 14 accessions which represented the main varieties cultivated in Spanish fields. Individual data related to plant, fruit, seed and phenology have been analysed using a multivariate analysis. This analysis showed intra- and inter-varietal diversity, as emphasized the most discriminant morphological traits in order to define similarities. Villaconejos accessions were morphologically distinct from the reference accessions (RA). Only the landraces belonging to Piel de Sapo market class showed common morphological affinities with the RA, grouping with them. The majority of Villaconejos traditional varieties clustered in five different groups, with no RA within, showing distinctive morphological singularities not described previously. Two of these groups showed some particularities in fruit traits, which are appreciated as quality marks for Spanish consumers. These results indicate that these accessions must be conserved as valuable genetic resources to enrich the Inodorus genetic bases for future breeding proposes worldwide. Furthermore, it should be considered the opportunity of promoting their cultivation under the shelter of a Protected Geographical Indication as a high quality melon. Finally, the discovering of such a high variability presented in a very small area, gives a clue for focusing, with an elevated probability of success, future surveys in similar ancestral European villages which in the past times, also supplied more populated cities with their farming products.     

Agromorphological genetic diversity of Spanish traditional melons

The variability of 62 Spanish landraces (and two hybrids used as reference) was described by analysing 39 morphological traits and eight SSR makers. Results showed that 81% of the examined genetic pool belonged to the inodorus type. Spanish traditional melons presented fruits from flattened and globular shapes to elliptical. Rind colour varied from pale green, almost white and yellow to dark green and almost black. Rind texture varied from smooth to intensely wrinkled. Spanish landraces also had larger fruits (average fruit weight ~2.6 kg) and longer vegetative cycles (117 days to maturity) compared to landraces from other geographical origins. Farmers seem to have focused on selection towards large fruits, which usually requires the longest production cycles. Fruit colour, size and shape seemed to have been determinant in varietal selection. Hierarchical clustering resulted in two main groups (climacteric and non-climacteric). The largest group was composed of 60 accessions of non-climacteric types, which includes the most demanded by national markets, ‘Piel de Sapo’, which fruits were characterized by an ovate or elliptical shape, a green rind, big spots and stains distributed over the whole fruit, a rounded blossom end shape and a very pointed stem end shape. The study demonstrates that the Spanish genetic pool is much more diverse. Wide variability was found in a geographical area with vast historical importance in melon farming. These evaluations has allowed the identification of several uniform groups of non-climacteric cultivars (‘Piel de Sapo’, ‘Mochuelo’, ‘Tendral’, Yellow/White, Winter and Black groups) and a set of highly variable climacteric ones. However, many accessions with singular properties remain unclassified, demonstrating the morphological variability of the studied collection. Melons in Spain have wide variability together with a vast historical importance on farms. Some fruit types, or at least some morphological characters reflected by painters during centuries, have reached the present. The present study confirmed the need to preserve these irreplaceable genetic resources and continue their study and evaluation for valuable traits which could enhance farmer’s opportunities for entering new markets.     

Diversity among landraces of Indian snapmelon (<Emphasis Type="Italic">Cucumis melo</Emphasis> var. <Emphasis Type="Italic">momordica</Emphasis>)

Diversity among 36 snapmelon landraces, collected from 2 agro-ecological regions of India (9 agro-climatic sub-regions), was assayed using RAPD primers, morphological traits of plant habit and fruit, 2 yield-associated traits, pest and disease resistance and biochemical composition (TSS, ascorbic acid, titrable acidity). Typical differences among accessions were observed in plant and fruit characteristics and snapmelon germplasm with high titrable acidity and possessing resistance to downy mildew, Cucumber mosaic virus, Zucchini yellow mosaic virus, Papaya ringspot virus, Aphis gossypii and Meloidogyne incognita was noticed in the collection. RAPD based grouping analysis revealed that Indian snapmelon was rich in genetic variation and region and sub-region approach should be followed across India for acquisition of additional melon landraces. Accessions of var. agrestis and var. momordica clustered together and there was a separate cluster of the accessions of var. reticulatus. Comparative analysis of the genetic variability among Indian snapmelons and an array of previously characterized reference accessions of melon from Spain, Israel, Korea, Japan, Maldives, Iraq, Pakistan and India using SSRs showed that Indian snapmelon germplasm contained a high degree of unique genetic variability which was needed to be preserved to broaden the genetic base of melon germplasm available with the scientific community. N. P. S. Dhillon and Ranjana contributed equally to this work and are considered the first authors.     

Agro-morphological diversity scenario in upland rice germplasm of Jeypore tract

The Jeypore tract of Orissa is one of the secondary centers of origin of cultivated rice. Recent exploration to that tract has resulted in a collection of 120 accessions of upland rice germplasm. They have been conserved in both National active collection and base collection centres for further utilisation as and when it will be required. Also a historical perspective on the genetical diversity of rice germplasm of the region has been highlighted. The characterization data revealed enormous morphological as well as agronomical variability among the landraces.