Penelope J. Bebeli

Cereal landraces for sustainable agriculture. A review

Modern agriculture and conventional breeding and the liberal use of high inputs has resulted in the loss of genetic diversity and the stagnation of yields in cereals in less favourable areas. Increasingly landraces are being replaced by modern cultivars which are less resilient to pests, diseases and abiotic stresses and thereby losing a valuable source of germplasm for meeting the future needs of sustainable agriculture in the context of climate change. Where landraces persist there is concern that their potential is not fully realised. Much effort has gone into collecting, organising, studying and analysing landraces recently and we review the current status and potential for their improved deployment and exploitation, and incorporation of their positive qualities into new cultivars or populations for more sustainable agricultural production. In particular their potential as sources of novel disease and abiotic stress resistance genes or combination of genes if deployed appropriately, of phytonutrients accompanied with optimal micronutrient concentrations which can help alleviate aging-related and chronic diseases, and of nutrient use efficiency traits.We discuss the place of landraces in the origin of modern cereal crops and breeding of elite cereal cultivars, the importance of on-farm and ex situ diversity conservation; how modern genotyping approaches can help both conservation and exploitation; the importance of different phenotyping approaches; and whether legal issues associated with landrace marketing and utilisation need addressing. In this review of the current status and prospects for landraces of cereals in the context of sustainable agriculture, the major points are the following: (1) Landraces have very rich and complex ancestry representing variation in response to many diverse stresses and are vast resources for the development of future crops deriving many sustainable traits from their heritage. (2) There are many germplasm collections of landraces of the major cereals worldwide exhibiting much variation in valuable morphological, agronomic and biochemical traits. The germplasm has been characterised to variable degrees and in many different ways including molecular markers which can assist selection. (3) Much of this germplasm is being maintained both in long-term storage and on farm where it continues to evolve, both of which have their merits and problems. There is much concern about loss of variation, identification, description and accessibility of accessions despite international strategies for addressing these issues. (4) Developments in genotyping technologies are making the variation available in landraces ever more accessible. However, high quality, extensive and detailed, relevant and appropriate phenotyping needs to be associated with the genotyping to enable it to be exploited successfully. We also need to understand the complexity of the genetics of these desirable traits in order to develop new germplasm. (5) Nutrient use efficiency is a very important criterion for sustainability. Landrace material offers a potential source for crop improvement although these traits are highly interactive with their environment, particularly developmental stage, soil conditions and other organisms affecting roots and their environment. (6) Landraces are also a potential source of traits for improved nutrition of cereal crops, particularly antioxidants, phenolics in general, carotenoids and tocol in particular. They also have the potential to improve mineral content, particularly iron and zinc, if these traits can be successfully transferred to improved varieties. (7) Landraces have been shown to be valuable sources of resistance to pathogens and there is more to be gained from such sources. There is also potential, largely unrealised, for disease tolerance and resistance or tolerance of pest and various abiotic stresses too including to toxic environments. (8) Single gene traits are generally easily transferred from landrace germplasm to modern cultivars, but most of the desirable traits characteristic of landraces are complex and difficult to express in different genetic backgrounds. Maintaining these characteristics in heterogeneous landraces is also problematic. Breeding, selection and deployment methods appropriate to these objectives should be used rather than those used for high input intensive agriculture plant breeding. (9) Participatory plant breeding and variety selection has proven more successful than the approach used in high input breeding programmes for landrace improvement in stress-prone environments where sustainable approaches are a high priority. Despite being more complex to carry out, it not only delivers improved germplasm, but also aids uptake and communication between farmers, researchers and advisors for the benefit of all. (10) Previous seed trade legislation was designed primarily to protect trade and return royalty income to modern plant breeders with expensive programmes to fund. As the desirability of using landraces becomes more apparent to achieve greater sustainability, legislation changes are being made to facilitate this trade too. However, more changes are needed to promote the exploitation of diversity in landraces and encourage their use.

Direct amplification of minisatellite-region DNA with VNTR core sequences in the genus Oryza

Abstract A polymerase chain reaction (PCR) application, involving the directed amplification of minisatellite-region DNA (DAMD) with several minisatellite core sequences as primers, was used to detect genetic variation in 17 species of the genus Oryza and several rice cultivars (O. sativa L.). The electrophoretic analysis of DAMD-PCR products showed high levels of variation between different species and little variation between different cultivars of O. sativa. Polymorphisms were also found between accessions within a species, and between individual plants within an accession of several wild species. The DAMD-PCR yielded genome-specific banding patterns for the species studied. Several DAMD-PCR-generated DNA fragments were cloned and characterized. One clone was capable of detecting multiple fragments and revealed individual-specific hybridization banding patterns using genomic DNA from wild species as well as rice cultivars. A second clone detected only a single polymorphic locus, while a third clone expressed a strong genome specificity by Southern analysis. The results demonstrated that DAMD-PCR is potentially useful for species and genome identification in Oryza. The DAMD-PCR technique also allows for the isolation of informative molecular probes to be utilized in DNA fingerprinting and genome identification in rice.

Collection, evaluation and classification of Greek populations of faba bean (Vicia faba L.)

Fifty-five Greek Vicia faba L. populations, collected from diverse areas, were planted at two dry and low fertility sites for evaluation and classification. Yield evaluation, which was carried out by Principal Component Analysis (PCA) on the basis of seven yield traits, showed the number of pods per plant, number of ovules and seeds per pod, and branching from the basal nodes to be the most important traits for population evaluation regarding yield. For population classification, four dissimilarity coefficients (Manhattan, Average Taxonomic Distance, Euclidean distance and squared Euclidean distance) and four multivariate methods (PCA, UPGMA, Neighbor-joining and Principal Coordinate Analysis) were evaluated using fifteen morphological and seven yield traits. Neighbor-joining was chosen as the most suitable multivariate method. This method combined with PCA for the seven yield traits, placed the populations into six groups. As revealed by the application of PCA on all twenty-two traits the grouping was based mainly on pod characteristics, stem thickness, plant height, 1000 seed weight and branching from basal nodes. Based on the results of the present study, a model is proposed for conserving cross-pollinated species, such as faba bean.

Repetitive, genome-specific probes in wheat (Triticum aestivum L. em Thell) amplified with minisatellite core sequences

The detection and analysis of DNA polymorphisms in crops is an essential component of marker-assisted selection and cultivar identification in plant breeding. We have explored the direct amplification of minisatellite DNA by PCR (DAMD-PCR) as a means for generating DNA probes that are useful for detecting DNA polymorphisms and DNA fingerprinting in wheat. This technique was facilitated by high-stringency PCR with known plant and animal minisatellite core sequences as primers on wheat genomic DNA. The products of DAMD-PCR from Triticum aestivum, T. durum, T. monococcum, T. speltoides and T. tauschii showed a high degree of polymorphism and the various genomes could be identified. Cloning of the DAMD-PCR products and subsequent Southern hybridization frequently revealed polymorphic probes showing a good degree of genome specificity. In addition, polymorphic, single locus, and moderately dispersed PCR products were cloned that may have a potential for DNA fingerprinting. Our experiments were limited primarily to diploid wheats and the results indicated that DAMD-PCR may isolate genome-specific probes from wild diploid wheat species that could be used to monitor genome introgression into hexaploid wheat.

Fruit cuticle lipid composition and water loss in a diverse collection of pepper (Capsicum)

Pepper (Capsicum spp.) fruits are covered by a relatively thick coating of cuticle that limits fruit water loss, a trait previously associated with maintenance of postharvest fruit quality during commercial marketing. To shed light on the chemical-compositional diversity of cuticles in pepper, the fruit cuticles from 50 diverse pepper genotypes from a world collection were screened for both wax and cutin monomer amount and composition. These same genotypes were also screened for fruit water loss rate and this was tested for associations with cuticle composition. Our results revealed an unexpectedly large amount of variation for the fruit cuticle lipids, with a more than 14-fold range for total wax amounts and a more than 16-fold range for cutin monomer amounts between the most extreme accessions. Within the major wax constituents fatty acids varied from 1 to 46%, primary alcohols from 2 to 19%, n-alkanes from 13 to 74% and triterpenoids and sterols from 10 to 77%. Within the cutin monomers, total hexadecanoic acids ranged from 54 to 87%, total octadecanoic acids ranged from 10 to 38% and coumaric acids ranged from 0.2 to 8% of the total. We also observed considerable differences in water loss among the accessions, and unique correlations between water loss and cuticle constituents. The resources described here will be valuable for future studies of the physiological function of fruit cuticle, for the identification of genes and QTLs associated with fruit cuticle synthesis in pepper fruit, and as a starting point for breeding improved fruit quality in pepper.

PCR primed with minisatellite core sequences yields DNA fingerprinting probes in wheat

Abstract Four minisatellite core sequences were used as primers in a polymerase chain reaction (PCR) technique, known as the directed amplification of minisatellite-region DNA (DAMD), to detect polymorphisms in three pairs of hexaploid/tetraploid wheat cultivars. In each pair, the tetraploid cultivar (genomic formula AABB) was extracted from its corresponding hexaploid (genomic formula AABBDD) parent. Reproducible profiles of the amplified products revealed characteristic bands that were present only in the hexaploid wheats but not in their extracted tetraploids. Some polymorphisms were observed among the hexaploid cultivars. Twenty-three DAMD-PCR amplified fragments were isolated and screened as molecular probes on the genomic DNA of wild wheat species, hexaploid wheat and triticale cultivars. Subsequently, 8 of the fragments were cloned and sequenced. The DAMD-PCR clones revealed various degrees of polymorphism among different wild and cultivated wheats. Two clones yielded individual-specific DNA fingerprinting patterns which could be used for species differentiation and cultivar identification. The results demonstrated the use of DAMD-PCR as a tool for the isolation of informative molecular probes for DNA fingerprinting in wheat cultivars and species.

Genetic diversity of Greek Aegilops species using different types of nuclear genome markers.

Random Amplified Polymorphic DNA (RAPD) and Inter-Simple Sequence Repeat (ISSR) analyses were used to evaluate genetic variability and relationships of Greek Aegilops species. Thirty-eight accessions of seven Greek Aegilops species [Ae. triuncialis (genome UC), Ae. neglecta (UM), Ae. biuncialis (UM), Ae. caudata (C), Ae. comosa (M), Ae. geniculata (MU) and Ae. umbellulata (U)] as well as Triticum accessions were studied. Nineteen RAPD and ten ISSR primers yielded 344 and 170 polymorphic bands, respectively, that were used for the construction of dendrograms. Regardless of the similarity coefficient and marker type used, UPGMA placed 38 Aegilops accessions into one branch while the other branch consisted of wheat species. Within the Aegilops cluster, subgroups were identified that included species that shared the same genome or belonged to the same botanical section. Within the Triticum cluster, two robust subgroups were formed, one including diploid wheat and another including polyploid wheat. In conclusion, results showed that there is genetic diversity in the Greek Aegilops species studied, and clustering based on genetic similarities was in agreement with botanical classifications.

Cultivar identification in upland cotton using RAPD markers

Using the random amplified polymorphic DNA (RAPD) method, the identification and the genetic description of 28 upland cotton (Gossypium hirsutum L.) cultivars currently cultivated in Greece was attempted. Based on the results of a preliminary experiment using 50 ten-base arbitrary primers, 24 were selected for the main experiment. DNA bands totaling 181 were observed, 118 (65.2%) of which were polymorphic. On the average, 7.5 DNA bands were amplified per primer, 4.9 of which were polymorphic. The unique identification of all cultivars studied was made possible using 27 specific polymorphic bands (markers) corresponding to 16 primers and a specially constructed key. The genetic similarity of the cultivars was estimated using Jaccards similarity coefficient, which ranged from 0.614 to 0.922, indicating a relatively narrow genetic base. Cluster analysis by the Unweighted Pair Group Method of Arithmetic means (UPGMA) showed that 21 of the cultivars could be placed into 3 major groups. A similar clustering of the cultivars was obtained using principal coordinate analysis.

Plant genetic resources of Lemnos (Greece), an isolated island in the Northern Aegean Sea, with emphasis on landraces

Lemnos Island is located in the Northern Aegean Sea and presents a rich biodiversity in plant genetic resources including wild species and crop landraces. Landraces have been cultivated under traditional agricultural systems and many have survived genetic erosion. They are mainly conserved in home gardens or in small fields, usually by elderly people, and are limited to local consumption. Two collecting expeditions were organized in Lemnos, a minor one in 2009 and a larger one in 2010. The results of the two current expeditions are discussed and compared with previous expeditions in the island. An overview is given on the crops cultivated, the landraces collected and those considered as lost, alongside with some information about their traditional uses. Finally, the importance of landraces in modern agriculture and preservation policies in Lemnos are discussed.

Plant regeneration and somaclonal variation from cultured immature embryos of sister lines of rye and triticale differing in their content of heterochromatin

SummaryPlants were regenerated from cultured immature embryos of two pairs of sister lines of triticale (X Triticosecale) cvs Rosner and Drira and five sister lines of rye (Secale cereale). The triticale lines differ in heterochromatic content of a particular rye chromosome (6R or 7R), while the rye lines differ in only one heterochromatic band. Variation in morphogenetic response was present between the triticale cultivars and between the rye lines. One of the rye lines (7RL+ +) showed a distinctive superior response in terms of somatic embryogenesis. These findings are discussed in relation to factors affecting morphogenetic response and genetic stability in culture.