Effect of seed storage on germplasm integrity of meadow fescue (Festuca pratensis Huds.)

Qualitative and quantitative changes in populations of meadow fescue (Festuca pratensis Huds.) after seed storage under different conditions were estimated. Phenotypic characters of plants grown from seeds stored for 54 months under two different storage conditions were compared. The germination of most of the seed accessions stored under favourable conditions remained at the initial level while storage under unfavourable conditions resulted in a decrease of the initial germination percentage as well as in an increase in the number of plants showing chlorophyll aberrations. There is great variability among the accessions resulting from storage under unfavourable conditions. In case of a few accessions a decrease in germination below 35% resulted in chlorophyll aberrations, faster flowering, shorter inflorescence and reduced viability of pollen grains.     

Origin of some minor vegetatively propagated Allium crops studied with RAPD and GISH

Three vegetative crops of sect. Cepa in genus Allium (Top onion, French grey shallot and viviparous triploid onion) of suspected hybridogenic origin were studied with genomic in situ hybridization (GISH) and random amplified polymorphic DNA (RAPD) markers. Related wild and cultivated species were included in the analysis in order to assess their contributions to the genomes of the investigated species. In A. × proliferum, the parental chromosomes derived from A. fistulosum and A. cepa were unequivocally identified by GISH, proving the hybrid status of this crop. The French grey shallot proved to belong to A. oschaninii according to the RAPD analysis and the GISH results, it is clearly separate from the normal shallots of A. cepa var. aggregatum. Thus the grey shallots are a new crop species and can be considered as an aggregatum form of the wild progenitor species A. oschaninii. The triploid viviparous onion comprises mostly A. cepa derived DNA in its genome. A non-cepa component could not be attributed to any of the sect. Cepa species included in this study and is most likely derived from one of the species of this section not yet identified by molecular means.     

The Garlic and Shallot Core Collection image database of IPK presenting two vegetatively maintained crops in the Federal ex situ genebank for agricultural and horticultural crops at Gatersleben, Germany

Garlic and shallot are important vegetable and spice plants, garlic is also a medicinal crop widely used throughout the world. Both belong to the genus Allium, which has been a main target of the taxonomical research at the IPK for many years. Therefore, the IPK’s living collection of Allium is one of the world’s largest special collections of this genus with garlic and shallot being essential parts of this collection. Furthermore, they are subject to special activities to preserve the material in vitro and in cryopreservation. Both methods help protecting valuable germplasm from abiotic and biotic threats and reduce the maintenance costs in the long term. At present, the garlic collection maintained at the IPK incorporates germplasm of 509 accessions in total including all safety duplicates not offered for exchange. Of shallot, 114 accessions are present as well. Passport data as well as phenotypic data of 159 accessions of garlic, one of great headed garlic and 16 of shallot are included in the Garlic and Shallot Core Collection Database (GSCC). An additional part of the database is a comprehensive collection of images illustrating morphological characteristics of the accessions. Exploration of the information is supported by a web-based application. The GSCC is available at http://www.ipk-gatersleben.de/databases/gscc.     

To what extent are threatened European plant species conserved in seed banks?

The number of ex situ conservation facilities has grown dramatically in recent years, and they have become increasingly integrated under national and regional conservation initiatives. However, little information is available about the extent to which plant species are appropriately represented in ex situ collections. This paper assesses whether seed/spore collections stored in European seed banks cover or not, and to what extent, the species currently threatened with extinction in the wild. Although a substantial amount of the European flora (ca. 70%) is currently stored in seed banks, we highlight the relatively poor representation of threatened species: only 27% of the taxa listed on the European threatened plant list and 44% of the taxa listed in Annex II of the EU Habitat Directive are stored in European seed banks. Some taxonomical groups most at risk, e.g. Pteridophytes and Orchidaceae, are also under-represented in European seed banks. By examining the number of accessions per species and the number of seeds per accession, this study also gave some insight on how well species are conserved, considering that these two variables are surrogates for genetic diversity. We have highlighted that at least two thirds of the threatened species stored in European seed banks likely suffer from too low genetic diversity in the collections. These analyses were essential to identify those collections and additionally the standards needed to maximize the usefulness of future collections.     

On the origin and distribution of normal cytoplasm of onion

Seventy populations of bulb onion (Allium cepa) from the Middle East, Central Asia, and Indian subcontinent introduced into the United States National Plant Germplasm System before 1964 were evaluated for the presence of normal (N) male-fertile and male-sterile (S) cytoplasms using three polymorphisms in the chloroplast genome. One accession acquired in 1948 from Turkey possessed only S cytoplasm. Fifty accessions possessed only N cytoplasm. Nineteen accessions possessed both N and S cytoplasm, of which S predominated in two accessions. The prevalence of N cytoplasm among populations originating from the Central Asian center of origin supports it as the wild type cytoplasm of onion. One accession of A. vavilovii recently collected in Turkmenistan was identical to N cytoplasmic onion for all of 42 previously characterized polymorphisms in the chloroplast genome, at all sites for 12 restriction enzymes in the nuclear 45s ribosomal DNA, and for 68 of 72 mitochondrial probe-enzyme combinations. Polymorphisms between this accession of A. vavilovii and three N cytoplasmic onions were detected only with cytochrome oxidase subunit 1 and four restriction enzymes. The cytoplasm of A. vavilovii is closely related to N cytoplasm and this species may be the progenitor of bulb onion.     

Preservation of seed viability during 25 years of storage under standard genebank conditions

Maintaining sufficient viability is critical to the sustainability of ex situ conserved seed collections. For this reason, accessions are regenerated when viability falls below a predefined threshold. Viability is monitored by determining the germination ability of accessions at predefined time intervals. Optimizing the frequency of these germination tests, in order to avoid waste of resources, is hampered by the scarce availability of data about seed longevity, particularly for material maintained under genebank conditions. Here we report on the analysis of nearly 40,000 germination test results collected for a wide range of crop species over a 25-years period by the centre for genetic resources, the Netherlands (CGN), where seeds of genebank accessions are dried to 3–7 % moisture content and stored for the long term under near-vacuum in aluminium foil bags at ?20 °C. The results indicate that seed viability is well maintained for the large majority of seed lots during the first 25 years after regeneration as only 3.3 % of the monitoring tests revealed below-threshold germination values. It is argued that the majority of these sub-standard seed lots are due to other causes than seed ageing, including dormancy problems and estimation error in germination testing. For material, maintained under the seed management procedures and storage conditions practiced by CGN, it is therefore recommended to delay the first germination monitoring tests to 25 years after regeneration.     

Genetic variability evaluation in a Moroccan collection of barley, Hordeum vulgare L., by means of storage proteins and RAPDs

The genetic variation existing in a set of barley (Hordeum vulgare L.) landrace samples recently collected in Morocco was estimated. Two kinds of genetic markers, seed storage proteins (hordeins) and random amplified polymorphic DNA (RAPD), were used. Only six out of 31 landraces were subjected to RAPD analysis. Both kinds of markers, RAPD and storage proteins, yielded similar results, showing that the level of variation observed in Moroccan barley was high: all landraces showed variability; 808 different storage protein patterns (multilocus associations) were observed among 1897 individuals (2.32 seeds per association, on average) with an average of 43 multilocus associations per accession. In general, genetic variation within accessions was higher than between accessions. The 100 polymorphic RAPD bands generated by 21 effective primers were able to generate enough patterns to differentiate between uniform cultivars and even between individuals in variable accessions. One of the aims of this work was to compare the effectiveness of RAPD versus storage protein techniques in assessing the variability of genetic resource collections. On average hordeins were more polymorphic than RAPDs: they showed more alternatives per band on gels and a higher percentage of polymorphic bands, although RAPDs supply a higher number of bands. Although RAPD is an easy and standard technique, storage protein analysis is technically easier, cheaper and needs less sophisticated equipment. Thus, when resources are a limiting factor and considering the cost of consumables and work time, seed storage proteins must be the technique of choice for a first estimation of genetic variation in plant genetic resource collections.     

Viability of Oryza sativa L. seeds stored under genebank conditions for up to 30 years

Germination ability, equilibrium relative humidity (eRH), and moisture content of ‘control’ seed samples representing 183 rice accessions stored in the active (2–4 °C) and base (?10 °C until 1993, then ?20 °C) collections of the T. T. Chang Genetic Resources Center were determined after storage for 20.5–30.5 years. Germination of seeds that had been stored in the base collection was generally high (>70 %), whereas germination was more variable for seeds stored in the active collection. Samples with lower viability after storage in the active collection were likely to have lower viability after storage in the base collection. There were significant differences in the moisture content-eRH relationship of the seeds depending on whether the seeds had been stored in the active or base collection. Based on re-test data for regular seed samples regenerated in 1979–1980 and stored in the active collection for up to 31 years, estimates of the time for ability to germinate to fall to 50 % (p ₅₀) ranged from 54 to 997 years. For the same seed samples stored in the base collection for approximately 31 years, ability to germinate has been maintained and germination increased due to improved procedures. The ability to germinate of base collection samples was also generally higher than that of ‘safety duplicate’ samples of the same seed lots that had been sent to the National Center for Genetic Resources Preservation, USA in 1981 and stored at ?18 °C. This may have been due to uptake of moisture either during processing for dispatch or as a consequence of poor packaging material. The results are discussed in relation to long-term seed storage and genebank management.     

Proteomic analysis of high protein soybean (Glycine max) accessions demonstrates the contribution of novel glycinin subunits

Limited biochemical information is available on soybean accessions that have seed protein content greater than 45% of the seed dry weight. SDS-PAGE analysis of seed proteins from nine soybean accessions revealed significantly higher amount of seed storage proteins in these accessions when compared with that of soybean cultivar Williams 82. High-resolution two-dimensional gel electrophoretic analysis of seed proteins revealed significant differences among several seed storage protein components in these accessions. A total of 51 protein spots were identified using peptide mass fingerprinting (MALDI-TOF MS). The contribution of these proteins to the overall protein content of the accessions was quantified using Delta2D image analysis software. Results showed that among the majority of the nine accessions, the largest difference in higher protein quantity was within the seed 11S storage globulins. The high protein trait from PI407788A was successfully transferred to an experimental line, LG99-469, demonstrating that this trait was transferable and robust.     

A multi-species assessment of genetic variability in Nigerian Amaranthus accessions: potential for improving intra- and interspecies hybridization breeding

Amaranthus yields could benefit from our growing understanding of the genetic variability within and among individual species of local germplasm collections. Thirty-seven Amaranthus accessions representing three different species were evaluated for seed yield and yield components across two seasons. The objectives were to characterize the genetic variability of seed yield and its components across species and determine relationships among seed yield and its components by phenotypic correlation. Significant differences (P < 0.01) existed for all the evaluated traits among the accessions. Characters such as seed yield, stem girth and leaf length showed higher heritability estimates in A. cruentus (14) than in A. hypochondriacus (12) and A. dubius (11) accessions. Seed yield was significantly and positively correlated with stem girth and thousand seed weight. However, it was significantly and negatively correlated with number of days to flowering, plant height, leaf number, number of branches. The clustering of the accessions, which was partially in agreement with species grouping, will help in the identification of diverse parents for use in the breeding program. Enormous amount of genetic variability found in seed yield and its components among Nigerian Amaranthus accessions will offer opportunity for grain and leaf yield improvement through breeding.     

Analysis of germplasm distribution patterns for collections held at the North Central Regional Plant Introduction Station, Ames, Iowa, USA

Understanding the patterns of distribution of plant genetic resources, especially the extent and contextual bases of distributions, may be critical in setting appropriate targets for seed multiplication, packaging, storage space, and other technical operations. We analyzed germplasm distribution patterns over a 12-year period for 10 crop collections conserved by the North Central Regional Plant Introduction Station in Ames, Iowa, to determine if distribution rates over a given time interval help predict future distributions and to document how distribution patterns vary among accessions within collections. We demonstrated that, with an appropriate tracking system and commonly available statistical software, germplasm distribution patterns can be easily analyzed and plotted over time. Data measured over periods of up to 3 years had little predictive value, while a 6-year period gave relatively accurate projections of future distributions. Patterns of distributions within collections varied between those that are approximately normally distributed and those best described by an exponential function, with larger collections tending to be non-normally distributed. Means and standard deviations of standardized, long-term distribution rates, calculated from samples of 200–700 accessions, accurately described the distributional rates of 90–95% of all accessions. The documentation of changes in usage patterns within and among collections as they mature is also discussed. Analysis of average shipment size suggests that germplasm distributions became more focused over time for 8 of the 10 collections analyzed. This may result when users request germplasm based upon knowledge about specific accessions gained through personal experience and by examining evaluation and characterization data.     

Incidence and viability of Acremonium endophytes in tall fescue accessions from North Spain

The tall fescue (Festuca arundinacea Schreb.) seed collection of the Centro de Investigaciones Agrarias de Mabegondo was surveyed to determine Acremonium infection rates. One hundred seeds from each of 19 accessions were microscopically examined for endophyte. Seventeen accessions (90%) contained endophyte with a range of 7 to 100%. Leaf sheaths of seedlings from the infected accessions were examined to determine the percentage of infection with viable endophyte. Only seven (41%) of the seventeen infected accessions collected and stored in 1995 at 4°C and 45–50% relative humidity contained viable endophyte with a range of 70 to 100%. While existing germplasm collections of Festuca arundinacea can provide only a small basis of Acremonium germplasm for study, newly collected Festuca arundinacea accessions will offer a greater diversity of Acremonium genotypes.     

Seed variability in <Emphasis Type="Italic">Madhuca longifolia</Emphasis> (Koenig) J. F. Macbride,a source of oil for use as bio-energy

Madhuca longifolia (Koenig) J. F. Macbride is native to India. Seeds contain 20–50% oil (Mahua oil) and the seed cake is used as manure. Mahua oil is used by rural communities for protection against storage pests. Almost all parts of this tree have medicinal properties. Four explorations and collection missions was conducted during July–October 2007 in Tamil Nadu and the Union Territory of Puducherry. This resulted in the collection of 55 accessions. Characterisation and analysis of 7 seed characters were done. The existence of enormous variability in seed length, thickness and 100 seed weight indicates scope for utilization of these accessions for selection of promising material for extraction of oil. Oil content of kernels has been evaluated. Kernel oil ranged from 44.43 to 61.50%. Three accessions IC556617 with 61.50, IC556632 with 60.80 and IC556632 with 60.55% of kernel oil are superior to the rest. Further collections in the site of these oil rich accessions are promising for more productive Madhuca longifolia genetic resources.     

Collection of Trifolium sp. and other forage legumes in Bulgaria

Joint collecting expeditions to improve the genetic diversity of Trifolium species in the U.S. National Plant Germplasm System (NPGS) were carried out in Bulgaria by scientists of the USDA-ARS, University of Florida, Texas A&M University, and Institute of Introduction and Plant Genetic Resources (IIPGR), Bulgaria, on 14–27 July 1990 and 30 July – 13 August 1993. The objectives were to collect seed of T. vesiculosum Savi and other annual clovers in southeastern Bulgaria in 1990 and seed of perennial Trifolium species in southwestern Bulgaria in 1993. A total of 246 collections were made of 50 Trifolium species (38 annual and 12 perennial) and 25 collections of 17 other forage legumes. Collections were made at 90 sites in 13 of the 20 floristic regions in Bulgaria. The genetic diversity of T. vesiculosum in the NPGS was increased by the addition of 16 accessions. Four Trifolium sp. were added to the NPGS. Land utilization for intensive grazing near villages and small plot farming in river valleys greatly increased between 1990 and 1993. Genetic erosion of the extensive Trifolium resources within Bulgaria may occur as these practices increase.     

Quercetin and isorhamnetin glycosides in onion (Allium cepa L.): varietal comparison, physical distribution, coproduct evaluation, and long-term storage stability

During onion processing, the outer dried protective layer (outer paper layer) and first two fleshy leaf layers are removed. This coproduct material is a potential commercial source of flavonoids especially quercetin. In the following study, the flavonoid composition was determined in coproduct materials and the press cake (material generated after juice extraction) in several commercially important onion varieties grown in California. Flavonoids were characterized and quantified using LC-(ESI)MS/MS and HPLC. The long-term stability of quercetin glycosides was assessed in dried coproduct materials stored at 4 and 22 °C over a 12 month period. In all varieties, the predominant forms of quercetin were the quercetin 3,4'-O-glucoside and 4'-O-glucoside. The first layer had significantly higher levels of flavonoids than the outer paper, second, and inner flesh layers on a DW basis (p < 0.05). Allium cepa "Milestone" contained the highest levels (p < 0.05) of flavonoids (1703 mg/100 g on a dry weight basis (DW). Onion press cake had significantly higher levels of total quercetin as compared with fresh onions (p < 0.05). The levels of 4'-O-glucoside significantly decreased during the first month of storage and remained stable for 12 months of storage at either 4 or 22 °C (p < 0.05).     

Lablab purpureus (L.) Sweet – diversity, potential use and determination of a core collection of this multi-purpose tropical legume

Two germplasm collections of Lablab purpureus L. Sweet totalling 249 accessions were grown in the field in Australia and Ethiopia and characterised using a common set of morphological and agronomic (M-A) attributes. Data from each site were analysed separately using multi-variate analysis and a classification constructed for each collection. There was considerable diversity within both collections, especially that maintained in Ethiopia. Time to flowering, seed weight, and plant height were the most important attributes in determining group allocation in both classifications. Both collections had a high proportion of L. purpureus subsp. purpureus, especially accessions similar to the Australian cultivars Highworth and Rongai. They also had good representation of the less common L. purpureus subsp. uncinatus Verdc. less common landraces and wild collections from either Africa or India. When combined, the two classifications provided an overview of diversity and highlighted the similarities and dissimilarities between the two collections. The vast range in plant types supported the view that lablab has the capacity to be a multi-purpose legume for both commercial and smallholder agriculture. Some important and less common germplasm identified were Ethiopian domesticated subsp. uncinatus, Ethiopian subsp. purpureus landraces collected from regional markets, semi-domesticated and wild accessions from southern Africa, and wild accessions from India. Using this overview, a core collection of germplasm was selected, which provides researchers with a sound basis for future plant breeding and agronomic studies with this important tropical legume.     

Morphological patterns in a world collection of <Emphasis Type="Italic">Cleome gynandra</Emphasis>

Cleome gynandra L. is a leafy vegetable native to sub-Saharan Africa and Asia and cultivated on dry areas. The plant plays an important role in the food and nutrition security of local communities. The objective of this study was to illustrate variation and diversity in the Cleome collection held at the World Vegetable Center. In total, 242 accessions were cultivated and morphologically characterized and analyzed. The gynophore and filament, both important organs of the flower, played a key role in taxonomical identification. High morphological variation was detected in traits including plant height, pod length, leaf size, flower color, and earliness. Distinct morphological differentiation was detected between Asian and African accessions. On average, the African accessions were larger, less uniform and later in flowering and seed maturation than the Asian accessions. The results were used to establish a core collection of 49 Cleome accessions. Gaps in the current collection were detected. High within-accessions diversity challenges the existing ex situ conservation system; to maintain diversity, seed should be collected from a large number of plants, both during the collection mission and in ex situ regeneration. The results are relevant for germplasm collection strategies and regeneration protocols for good genebank practices.     

Assembling a core collection from the flax world collection maintained by Plant Gene Resources of Canada

Between 1998 and 2008, numerous projects were conducted by the Canadian national genebank, Plant Gene Resources of Canada, for the regeneration, characterization and evaluation of the whole flax (Linum usitatissimum L.) germplasm collection. The whole collection comprised 3378 accessions and, according to the passport data, several of these were probably genetically very similar or even identical. Therefore, a subset of 381 accessions was selected that represented the diversity found in the whole collection. Sampling accessions from the whole collection was made using characterization and evaluation data and followed six different methods: (1) For seven qualitative characters, each unique combination of character expression was represented by three accessions; (2) for quantitative characters, a fixed number of accessions representing the lowest and highest observed values was included; (3) for stem fibre content, disease ratings, seed vigour and drought tolerance, a fixed number of accessions with desirable performance was included; (4) a subset of the 57 most distinct accessions based on RAPD markers was included; (5) a subset of 40 pure lines that were created based on extreme low and high values for 1000 seed weight, seed oil content and fatty acid profiles was included; (6) a subset of fibre flax cultivars of known relevance in European flax breeding and another subset of flax cultivars of known relevance for North American linseed breeding were included. The goal was to maximize the diversity available in a limited number of flax accessions by preserving the range of variation present in the whole collection, while improving evenness. The core collection was assembled in response to requests by flax breeders. This paper compares distribution parameters in the whole and core collections.     

Genotypic diversity and patterns of variation in a germplasm material of Ethiopian mustard (Brassica carinata A. Braun)

The amphidiploid species Brassica carinata A. Braun is believed to have originated in the plateaus of Ethiopia and has been cultivated there as an oilseed crop since antiquity. The species possesses agronomically important genes of rare occurrence. Although there is a large number of collections in Ethiopia, information on the extent of their genetic diversity is very limited. Thirty-six accessions of ecologically diverse regions were tested at three locations and multivariate analyses on 13 morphological and seed characters were performed. There was generally a large amount of divergence in all characters. Geographic isolation of genes, however, was not observed. Both principal component and cluster analyses disclosed complex relationships among the accessions and characters. Accessions with potential genes of interest to improve earliness, yield components and oil and protein contents have been identified. Length of growing period and yield components contributed most for divergence and clustering pattern. Oil, glucosinolate and protein contents also varied among the accessions but not so much between clusters. Increasing protein in high-glucosinolate genotypes reduced oil content and should be more efficient in genotypes of low-glucosinolate genetic background.