Variety registration is an important area of plant genetic resource characterization and utilization. Within the European Union, varieties must be included on a National List (NL) of a member state or on the Common Catalogue (a compilation of the NLs of the member states) before seed can be sold. This requires a series of tests and trials which assess if the variety is distinct, uniform and stable (DUS) and if it has sufficient value for cultivation and use (VCU). The same DUS criteria are also used world-wide for the granting of Plant Breeders' Rights (PBR), an intellectual property protection system. Both DUS and VCU are currently assessed primarily using field-based trials. However, the potential use of biochemical and molecular markers for DUS purposes is being actively investigated and such markers could have an important role to play in maintaining the quality and scope of PBR in an environment where the increasing number of countries involved in DUS testing and the number of variety comparisons to be made are causing logistical difficulties. More recently, given firstly the increase in the availability of markers from expressed regions of the genome, and secondly the possibilities raised by detailed DNA sequencing programmes and the association of markers (particularly single nucleotide polymorphisms, SNPs) with specific genes, the prospects for a more molecular approach to VCU are also being discussed. This paper reviews the current situation with regard to the use of molecular markers for DUS and VCU testing and considers future prospects for variety registration in the 21st century, ‘-omics’, era.

Increased amino acid content in high-oil maize (Zea mays L.) grain may add further value to its use in livestock rations, especially if this enhanced amino acid content is consistent across varying growing conditions. Most high-oil maize (HOM) grown in the USA utilizes the TopCross system which involves planting a blend (TC Blend) of two types of maize. Field experiments and on-farm studies were conducted in 1997 and 1998 to compare the amino acid profile of grain from HOM TC Blends with that of their normal-oil maize (NOM) counterparts across a range of production environments in Ohio. In 1997, the composition of four amino acids (lysine, methionine, glycine and arginine) was significantly higher in HOM compared to NOM grain. In 1998, nine amino acids (lysine, methionine, glycine, arginine, asparagine, threonine, serine, cysteine and tryptophan) were greater in HOM than in NOM grain. Lysine and methionine content in HOM grain averaged 12 and 13% higher than in NOM grain in both years. The number of amino acids significantly affected by the grain parent was greater than that for maize type each year. A significant maize type × grain parent interaction for a limited number of amino acids suggest that TC Blend grain parents may affect the consistency of amino acid composition in HOM grain. Results of this study demonstrate that the levels of several amino acids, including economically important lysine and methionine, were consistently greater in HOM than in NOM grain across a range of production environments. Modelling with livestock ration balancing software showed that the additional amino acids and oil in HOM added 12–20% to its value as livestock feed.

The concept of Intellectual Property Rights (IPRs) is a very old one, dating as far back as Ancient Greek times. A specific system of IPRs for living organisms was adopted internationally for the first time, for plant varieties, in 1961, through the UPOV Convention. Progress in the field of biotechnology in the 1970s was the starting point for extensive debate on the patenting of living material among both the scientific community and in public fora, resulting in several independent international agreements which, in turn, contributed to the evolution of the Patent System for gene protection and of the statute of plant genetic resources. But, in their current state, the provisions of these various agreements are not globally coherent and, more particularly, conflict specifically with plant varieties and genetic resources. The questions could be solved by generalizing the cornerstone principle of the UPOV Convention, i.e. the free use of genetic variability, which seems to meet the demand of the scientific community.

This paper reviews marker methods based on retrotransposons and illustrates examples from oil palm. Prior to this study, very little had been known about the repetitive DNA present in oil palm and no marker systems based on retrotransposons had been developed. Firstly, copia like retrotransposons of Elaeis oleifera, Elaeis guineensis and Cocos nucifera were characterized by performing phylogenetic analyses on a portion of the reverse transcriptase (RT) gene of copia-like retrotransposons. The results identified three classes of copia-like retrotransposons in the three species studied. While the C-class RT sequences seemed to have amplified preferentially only in Elaeis guineensis, the classes A and B were present in Elaeis guineensis and Cocos nucifera as well. Secondly, long terminal repeats (LTRs) from these retrotransposon classes were isolated as a prelude to developing a marker system in palm based on retrotransposons. The B- and C-class LTRs were isolated by primer walking from the RT region and the B-class from Elaeis oleifera and Cocos nucifera with the use of oil palm-specific RnaseH primers. The method has been developed and applied for breeding purposes in oil palm, hybrids between E. oleifera and E. guineensis and within the tribe Cocoeae. There is scope for the method to be used in the isolation of new retrotransposon families endogenous for each palm species and as an alternative to amplified fragment length polymorphism (AFLP) in diversity studies within the Palmae.

Mungbean seeds are often attacked by bruchid insects causing serious damage to the grain during storage. The susceptible/resistant characteristics of mungbean against bruchids have been investigated at the protein level. Seeds of the susceptible cultivar Osaka-ryokuto and the resistant near-isogenic line BC20 were collected at 5, 9, 13, 17, 21 and 23 days after flowering. Crude, cytosol and membrane protein fractions were extracted from the seeds for twodimensional polyacrylamide gel electrophoresis analysis. In Osaka-ryokuto six proteins were found in higher amounts compared to BC20 in the crude protein extract. Some of these proteins were homologous to legumin B and liguleless1. And in BC20 one unique protein, homologous to SalF1R, was identified. In the subcellular protein fractions, nine unique proteins were identified in BC20, seven in the cytosol and two in the membrane fractions. These proteins were homologous to chitinase, alanine aminotransferase in membrane, and SalF1R, β-1,3- glucanase, protein phosphatase 2A, peroxidase BP 2A, provicilin and canavalin in the cytosol fractions. Previous findings of many researchers reveal that these are mainly defence-related proteins. Thus, it is suggested that the above proteins may play a role in maintaining the resistance of line BC20.

Artichoke, Cynara cardunculus var. scolymus is a diploid outcrossing species, originated in the Mediterranean basin, which has been much appreciated both for its tasty heads and pharmaceutical properties since ancient times. The species includes two more botanical varieties: C. cardunculus var. altilis, the cultivated leafy cardoon and C. cardunculus var. sylvestris, the presumed wild progenitor of artichoke, which are completely interfertile with the cultivated globe artichoke and all together they form the primary gene pool of artichoke. The secondary gene pool includes at least seven wild Cynara species. A high level of morphological variation, essentially in head shape and size, is observed in artichoke varieties. Amplified fragment length polymorphism (AFLP) markers were used in order to assess genetic variation and relationships among artichoke varieties and between these and some of their wild relatives. A selected group of wild and cultivated artichoke accessions belonging to different clusters detected on a morphological basis and from various geographical origins was chosen for the analysis. Twenty-four primer combinations were initially tested to evaluate their ability to detect polymorphism between samples. Nine primer combinations were chosen for further analysis on 39 cultivated artichokes, two wild progenitors, one cultivated cardoon, one sample of C. cornigera, one of C. humilis and two samples of C. syriaca. A high level of polymorphism was observed for AFLP markers. The polymorphic bands obtained were scored and used to assess genetic similarity among wild and cultivated accessions and finally to construct a UPGMA dendrogram and principal co-ordinate (PCO) analysis based on Jaccard's similarity index. The artichoke wild progenitor was quite distantly related to the cultigen and occupied a separate branch in the dendrogram. However, wild C. cardunculus was more similar to the artichoke than were the other wild species, corroborating the idea that it is the wild progenitor of cultivated artichokes. Within the cultivated artichoke, the dendrogram derived from AFLP analysis produced branches which roughly corresponded to the groups obtained on the basis of morphological and physiological characteristics. The groups were homogeneous enough, except for the ‘Romaneschi’ types, which proved to be quite genetically variable, and did not cluster in a single branch. This is interpreted on the grounds of the possible selection pathway of this more modern morpho-group.

Six distinct market samples of Dacryodes edulis (safou/African plum) with different fruit colour were characterized in terms of morphological, sensory and nutritional traits that can guide the selection of high-value individual trees for cultivar development. The morphological characteristics assessed were: fruit length, fruit width, fruit and flesh weights, flesh thickness and pulp/fruit ratio, while the sensory characteristics were: acidity, aroma astringency, bitterness, oiliness, preference, fibrousness and sourness. In addition, the biochemical composition (carbohydrates, ash, protein, lipids, dietary fibres, vitamin C, iron, calcium and phosphorus contents) was determined. These attributes all showed great variability, suggesting their importance in the identification of multi-trait ideotypes for potential cultivar development.

In this study an evaluation was made of changes in the genetic variation of barley (Hordeum vulgare L.) of Nordic and Baltic origin, from the end of the 19th century until today. A comparison of Nordic and Baltic material with exotic material that has not been subjected to intense selection was also made. A total of 293 accessions, including 160 Nordic and 80 Baltic landraces, cultivars and breeding lines, and another 53 landraces of exotic origin (Central Asia), were surveyed using isozyme starch gel electrophoresis. For the four isozymes studied, 28 alleles at 11 loci were observed. The average total genetic diversity value (HT) for individual loci ranged between 0 and 0.519. In the exotic material nine loci were polymorphic and two monomorphic, compared to seven polymorphic and four monomorphic loci in the Nordic and Baltic material. Some of the rare alleles were detected only in the exotic material. The studied isozyme loci of Nordic and Baltic material indicated that modern cultivars have a lower average genetic diversity value compared to the landraces, old and exotic material.

We suggest a diversity-dependent strategy, based on Principle Component Analysis, for selecting distinct accessions/parents for breeding from a soybean germplasm collection comprising of 463 lines, characterized and evaluated for 10 qualitative and eight quantitative traits. A sample size of six accessions included all the three states, namely low, medium and high of the individual quantitative traits, while a sample of 16–19 accessions included all the 60–64 distinct states of qualitative as well as quantitative traits. Under certain assumptions, the paper also develops an expression for estimating the size of a target population for capturing maximum variability in a sample three accessions.

Research on resistance to black pod caused by Phytophthora megakarya has been carried out in Cameroon for 40 years. It has yielded a significant database which is currently being exploited for the modelling of disease resistance levels of the major clones selected as candidate parents for the creation of new resistant and consequently more productive cocoa varieties. These outputs are expected to meet the increasing need of producers for improved planting material to extend cultivation into emerging cocoa-growing areas or to renew old cocoa production basins. These outputs have been made possible through an international collaborative effort and, more recently, within the scope of regional or international initiatives sponsored by CFC/ICCO/IPGRI and by CAOBISCO. New selection approaches and methodologies, such as rapid field assessment and early screening tests, have been promoted, along with the use of molecular tools for genome mapping and disease resistance quantitative trait locus (QTL) identification. Promising material has been introduced, evaluated in the laboratory and on-farm, and compared to the local cocoa germplasm. The basic knowledge developed from this local database has enabled the definition of an empirical disease resistance sorting and subsequently a prediction of the yield gains expected from the use of different cocoa clones and possible combinations (progenies) available on-station and in farmers' stands in different agro-ecological growing conditions; or which have to be created following strategies in the new breeding programme.

Characterization of plant germplasm using molecular techniques is playing an increasingly important role in the management and utilization of plant genetic resources, but has its limitations in the screening of large numbers of accessions held in seed genebanks worldwide. Bulking individual plants from one accession or group to form a representative sample is a promising approach to widening the scope of a characterization, but it is not without technical problems in detecting genetic variation. This review was conducted to assess the technical pitfalls of bulking, and to evaluate the effectiveness of various bulking methods in the assessment of genetic variation and genetic relationships, and in the identification of plant germplasm. Clearly, some alleles, particularly those occurring at low frequency, may go undetected in a bulked sample, depending on the bulking methods and the molecular techniques used. As a result, genetic diversity estimates and genetic relationship inferences can be significantly biased. Germplasm identification may not be always reliable. Thus, it is imperative that the detection limit imposed by bulking be assessed for a newly initiated molecular germplasm characterization and bias be considered in interpretation of the resulting characterization data. Equally imperative is the need for continuous efforts of exploring efficient bulking procedures for the screening of large germplasm collections, particularly by the newly developed marker systems.

We have compared the genetic diversity of 24 Chinese weak-winter, Swedish winter and spring Brassica napus accessions by inter-simple sequence repeats (ISSRs). Using cluster analysis (UPGMA) based on 125 polymorphism bands amplified with 20 primers, the 24 accessions were divided into three groups. Six Swedish winter lines and eight Chinese weak-winter lines were in group I and group II consisted of two Chinese weak-winter lines, Xiangyou15 and Bao81. The third group contained eight Swedish spring lines. Principal coordinates (PCO) analysis showed similar groupings to cluster analysis. Results from cluster analysis and PCO analysis showed very clearly that Chinese weak-winter, Swedish spring and winter accessions were distinguished from each other and Chinese weak-winter accessions in this study were genetically closer to Swedish winter accessions than to Swedish spring accessions. The Chinese weak-winter accessions had larger diversity than the Swedish spring or winter accessions. This study indicated that ISSR is a suitable and effective tool to evaluate genetic diversity among rapeseed germplasm.

A set of 146 representative adzuki (Vigna angularis var. angularis and var. nipponensis) germplasm from six Asian countries with a tradition of adzuki bean production, together with an outgroup standard rice bean (Vigna umbellata), were analysed by amplified fragment length polymorphism (AFLP) methodology using 12 informative primer pairs. A total of 313 unambiguous polymorphic bands were created. According to the dendrogram, using cluster analysis based on AFLP banding, 143 of the accessions were distinct and revealed enough genetic diversity for identification and classification of accessions within Vigna angularis. A neighbour-joining tree was generated using the newly developed Innan's nucleotide diversity estimate from the AFLP data. From analysis, seven distinct evolutionary groups, named ‘Chinese cultivated’, ‘Japanese cultivated’, ‘Japanese complex-Korean cultivated’, ‘Chinese wild’, ‘China Taiwan wild’, ‘Nepal- Bhutan cultivated’ and ‘Himalayan wild’, were detected. Nucleotide diversity with geographical distribution of each group is discussed, regarding the evolutionary relationships between wild and cultivated adzuki beans. The preliminary results indicated that cultivated adzuki beans have been domesticated from at least four progenitors with at least three geographical origins.