Analysis of the genetic diversity of wild, Spanish populations of the species Elymus caninus (L.) Linnaeus and Elymus hispanicus (Boiss.) Talavera by PCR-based markers and endosperm proteins Rosa Nieto-López, Carlos Casanova, Consuelo Soler To cite this version: Rosa Nieto-López, Carlos Casanova, Consuelo Soler. Analysis of the genetic diversity of wild, Spanish populations of the species Elymus caninus (L.) Linnaeus and Elymus hispanicus (Boiss.) Talavera by PCR-based markers and endosperm proteins. Agronomie, EDP Sciences, 2000, 20 (8), pp.893-905. 10.1051/agro:2000166. hal-00886092 HAL Id: hal-00886092 https://hal.archives-ouvertes.fr/hal-00886092 Submitted on 1 Jan 2000 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Agronomie 20 (2000) 893–905 893 © INRA, EDP Sciences 2000 Original article Analysis of the genetic diversity of wild, Spanish populations of the species Elymus caninus (L.) Linnaeus and Elymus hispanicus (Boiss.) Talavera by PCR-based markers and endosperm proteins Rosa María NIETO-LÓPEZ, Carlos CASANOVA, Consuelo SOLER* Department of Plant Breeding and Biotechnology, SGIT-INIA, Finca “La Canaleja”, PO Box 1045, Alcalá de Henares, 28800 Madrid, Spain (Received 12 April 2000; accepted 3 July 2000) Abstract – The use of wild species in plant improvement is greatly favored if their genetic variability and taxonomical relationships are known. A study was made of the genetic variability of fifteen Spanish populations of E. caninus and E. hispanicus. The relationships among these species and E. panormitanus were also investigated. Intra- and interpop- ulational variation was determined by electrophoresis of endosperm proteins. DNA polymorphisms generated by PCR amplification using arbitrary and specific primers were used to determine interpopulational variability and interspecific relationships. The variability observed was similar to that reported for other autogamous species. The reduced variabil- ity observed in some populations of E. caninus was attributed to increased distance from the species center of distribu- tion and to the founder effect. Endosperm proteins and random amplified polymorphic DNAs (RAPDs) provided com- plementary results and differentiated most populations of E. caninus. The interspecific relationships observed were more in agreement with the classification of E. hispanicus as a different species to E. panormitanus than as a variant form of the same taxon. Elymus / genetic diversity / endosperm proteins / PCR-based markers Résumé – Analyse de la diversité génétique de populations sauvages espagnoles d’Elymus caninus (L.) et d’Elymus hispanicus (Boiss.) Talavera par des marqueurs PCR et des protéines de réserve. L’utilisation des espèces sauvages en amélioration végétale implique de connaître leur variabilité génétique et leurs relations taxonomiques. Une étude a été réalisée sur la variabilité génétique de quinze populations de E. caninus et E. hispanicus. Les relations entre ces espèces et E. panormitanus ont été ainsi examinées. La variation intra- et interpopulation a été déterminée par electrophorèse des protéines de réserve. Les polymorphismes d’ADN générés par amplification PCR à l’aide d’amorces aléatoires et Communicated by Nicolás Jouve (Madrid, Spain) * Correspondence and reprints [email protected] Plant Genetics and Breeding 894 R.M. Nieto-López et al. spécifiques ont été utilisés pour déterminer la variabilité intraspécifique et les relations interspécifiques. On a observé une variabilité similaire à celle décrite pour d’autres espèces autogames. Dans quelques populations de E. caninus, on a observé une réduction de la variabilité qui peut être attribuée à un éloignement du centre de distribution de cette espèce ainsi qu’à un effet de fondation. Les protéines de réserve et l’ADN polymorphe amplifié aléatoirement (RAPD) appor- tent des résultats complémentaires et différencient la plupart des populations de E. caninus. Les relations interspécifiques observées sont concordantes avec la classification de E. hispanicus comme espèce différente de E. panormitanus et non comme une forme variante du même taxon. Elymus / diversité genétique / protéines de réserve / marqueurs RAPD 1. Introduction found in E. scabrus (Löve) which is given the genomic formula SSYYWW [32]. According to the genomic classification of Löve Owing to the importance of wild species as a (1984), two representatives of this genus are source of genes for breeding cultivated species, known in Spain: E. caninus (L.) Linnaeus (also international organizations and seed banks are very known as Agropyron caninum (L.) P. Beauvois or interested in their collection and conservation. The Roegneria canina (L.) Nevski.) and E. panormi- effective use of wild species in plant improvement tanus (Parl.) Tzvelev (also called Agropyrum is greatly favored if their genetic variability [24] panormitanus Parl. var. hispanicus Boiss. or and the taxonomical relationships between them is Elymus hispanicus (Boiss.) Talavera). known. Elymus caninus has the genomic constitution According to the genomic classification of Löve SSHH and grows in Europe and Asia. In Spain, it (1984), Elymus is the largest genus of the tribe grows in the Cantabrian Range, the Pyrenees, Triticeae, with more than 150 perennial species. It Galicia (Northwest Spain) and the Central and is also the most cosmopolitan, growing in Europe, Iberian Ranges (Central Spain). Its southern limit Asia, North America, South America, New lies at 37º north. Elymus panormitanus (Parl.) Zealand and Australia [9]. Many of the species of Tzvelev is another tetraploid species with the Elymus have good fodder qualities, are highly pro- genomic constitution SSYY, and grows in Europe ductive in favorable conditions, and tolerate differ- and Asia [33]. In Spain it is found in the southern ent diseases. part of the peninsula [20]. The chromosome number of Elymus varies from The relationships among certain species of the 2n=4x=28 to 2n=8x=56. Some 75% of Elymus Elymus genus have been clarified using different species are tetraploid with the genomic constitution biochemical and molecular systems such as SSHH. The S genome comes from Pseudo- isozymes, DNA repeated sequences, microsatel- roegneria and the H from Critesion [9]. Asiatic lites and random amplified polymorphic DNA species such as E. ciliaris have a Y genome of (RAPD) markers. One variant of the polymerase unknown origin rather than the H genome, and are chain reaction (PCR) technique used to obtain mol- represented genomically as SSYY [18]. The major- ecular markers consists of amplifying DNA frag- ity of hexaploids are segmental autoploids ments previously localized in the genetic map of a particular species. This technique has been used in S1S1S2S2HH or SSH1H1H2H2, and in some cases such as E. drobovii their genomic formula is SSH- certain Triticeae, such us Triticum spp. and HYY [8]. Recently the P genome of Agropyron Hordeum spp. [6, 22, 30]. spp. Gaertn has been identified in E. kengii which The purpose of the present study was to analyze has the genomic formula SSYYPP [13]. Similarly, the genetic variability of fifteen Spanish popula- the W genome of Australopyrum (an endemic tions of E. caninus and E. hispanicus. The relation- genus of Australia and New Zealand) has been ships among these species and E. panormitanus Genetic diversity of Elymus Spanish populations 895 was also investigated using biochemical For a better understanding of the relationships (endosperm proteins) and molecular (PCR) tech- between the analyzed species, E. panormitanus niques. In the latter, primers with both arbitrary (PI531646 from Iraq), Pseudoroegeneria spicata sequences (RAPDs) and with specific sequences (PI537387 from the USA), and P. geniculata were used. (PI502271 from the Russian Federation), all pro- vided by the Western Regional PI Station USDA- ARS, and Hordeum chilense (H80 from Chile), 2. Materials and methods provided by the University of Córdoba (Spain), were included in the study. 2.1. Plant material 2.2. Protein analysis The plant material consisted of fifteen wild, Spanish populations belonging to the species Endosperm proteins were analyzed according to E. caninus and E. hispanicus (Tab. I). This material the method of Payne et al. [23] using polyacry- forms part of the living collection of wild relatives lamide gels in the presence of 10% SDS (SDS- of cultivated Triticeae obtained from natural envi- PAGE). In order to perform this analysis without ronments [25] which is maintained at the Plant interfering with germination, a piece of embryo- Breeding and Biotechnology Department of the free endosperm was separated out. In the majority I.N.I.A (La Canaleja, Madrid, Spain). In order to of cases 30 individuals per population were ana- conserve the original structure of the natural popu- lyzed. Individual samples were inserted into the lation, specimens were collected at random and gels. A molecular weight standard (Sigma 6H) was each sample obtained following the method of also used which contained myosin (205 Kd), galac- Hawkes [11]. About