De Ron, A. M.; A. P. Rodio; M. De la Fuente; M. Santalla. 2010. LEurope, un centre de diversification secondaire du haricot. En: A.

Jacobsen (Direction) Du fayor au mangetout. Lhistoire du haricot sans en perdre le fil, 30-33. ditions du Rouergue. Rodex, France

EUROPE: A SECONDARY CENTRE OF BEAN DIVERSIFICATION Antonio M. De Ron, A. Paula Rodio, Mara De La Fuente, Marta Santalla Misin Biolgica de Galicia (MBG), National Spanish Research Council (CSIC) Pontevedra, Spain ORIGIN AND EVOLUTION OF THE EUROPEAN BEANS The common bean (Phaseolus vulgaris L.) was domesticated in two distinct regions of the New World, in Mesoamerica and in the Andean region in South America (Gepts et al., 1986). The Mesoamerican genetic pool includes small seeded varieties, usually high yielding, while the Andean pool is characterized by medium to large seeded germplasm, but its productivity is lower than the Mesoamerican varieties. The divergence of the two major domesticated genetic pools is supported by studies of variability in plant and seed morphology, phaseolin (the major storage seed protein), isozymes and DNA (Gepts et al., 1986; Singh et al., 1991a; Singh et al., 1991b; Haley et al., 1994). In pre-Columbian times, between Mesoamerica and South America some limited bean germplasm exchange took place, but much-more extensive seed movement occurred after the 1500s. The common bean was introduced into the Iberian Peninsula (Spain and Portugal), from Central America around 1506 (Ortwin-Sauer, 1966) and from the southern Andes after 1532, by sailors and traders who brought with them the nicely coloured and easily transportable seeds as a curiosity (Debouck and Smartt, 1995). Some studies have provided evidences for the existence, in the Iberian Peninsula, of varieties of the Andean and Mesoamerican genetic pools that could have been disseminated through other countries, along the European Mediterranean basin, such as Greece, Cyprus and Italy, as indicated by the phaseolin types found in such areas (Lioi, 1989). In the last years, many of the bean landraces or traditional varieties, maintained in the germplasm collection at the Misin Biolgica de Galicia (MBG-CSIC), were studied with the goal of identify their genetic variation and their affiliation to the Mesoamerican or Andean genetic pools. These studies were based upon plant and seed morphology, phaseolin and isozyme profile and DNA (microsatellite) polymorphism. The result indicates that the level of genetic variation in the common bean has not been eroded since the introduction of its ancestors that came from the American centers of domestication to the Iberian Peninsula. An important feature of genetic diversity in the bean varieties from the Iberian Peninsula is the

existence of a large-seeded white Andean phenotypes, such as favada (figure 1), canellini and white kidney, and Mesoamerican phenotypes, such as great northern, large great northern and hook (figure 2) which have a combination of Mesoamerican and Andean genes. Therefore, they could constitute intermediate forms or recombinants between both genetic pools. Iberian landraces suggest interesting questions about the nature of the variation described, as well as the evolutionary forces affecting the current European common bean germplasm. The antique Andean populations, originated in latitude similar to the Iberian Peninsula, may have had a competitive advantage over genotypes of other origins because of a better photoperiod adaptation. The common bean promptly reached a wide diffusion in Europe (Zeven, 1997) and, over five centuries, the European farmers selected and maintained many different types of common bean varieties depending on their preferences and uses, which resulted in a divergent selective pressure. The wide differentiation of types currently observed within the European common bean varieties could be a result of that selective process. The current European common bean germplasm collections show a wide variation of phenotypes, often large and extra-large seeded, compared with the germplasm from the areas of origin in the Americas (Santalla et al., 2001; Rodio et al., 2001; Negri and Tosti, 2002). Occasional outcrossing, adaptation to particular environmental conditions (temperature, moisture, photoperiod, soil fertility, diseases and insects), cropping systems and strong selection of seed types for consumer preferences, might have played a significant role in the evolution of the new variation of the common bean in the Iberian Peninsula. Thus, new germplasm (favada, hook and large-great-northern varieties), that could be considered as Iberian forms, could have probably emerged from initial recombination events between the Mesoamerican and Andean gene pools, that are better adapted to the prevailing conditions in the Iberian Peninsula. The new Iberian forms could have, subsequently, been disseminated to other parts of Europe, particularly through the Mediterranean basin contributing, in this way, to much-wider variation observed in European germplasm (Lioi, 1989; Gil and De Ron, 1992; Zeven, 1997). Hence, the Iberian Peninsula, mainly the north and northwest regions, could be considered as a secondary center of genetic diversity for the common bean, especially the large white-seeded varieties (Santalla et al., 2002). During the process of secondary diversification three new groups of bean varieties, which will be described in the next sections, arose (Rodio et al., 2006): Mesoamerican extreme types, Mesoamerican recombinant types and Andean recombinant types (table 1).

Figure 1A. Favada (Faba Galaica) seed type.

Figure 1B. Favada type growing in a farmer field in the North of Spain (Lourenz, Lugo).

Figure 2. Hook (Gantxet) seed type.

NEW TYPES OF EUROPEAN BEANS Mesoamerican extreme types Large seeded bean varieties are preferred in Spain probably as a heritage of the preColumbian times, when large seeded faba bean (Vicia faba L.) was often cultivated. In fact, the "faba" and "haba" names are currently used in the North of Spain, referred to dry bean and "habichuela" (in Spanish it means 'small faba') which are common names for snap bean in Spain and in the Americas. Some extra-large white seeded Spanish types were recognized already by Puerta-Romero (1961) with different local names: Ganset, De cuadro, Valenciana, Monquili de enrame, De la sierra, and Blanca vinosa. Gepts and Bliss (1988) observed that some bean populations from France such as Flageolet blanc a longue cosse, Sabre Nain Etoile du Nord and Tarbais, had large seeds and Mesoamerican phaseolin type. In Italy it is also important the variety Fagiolina Arsolana, a large-seeded type with Mesoamerican phaseolin pattern. The Spanish Ministry of Agriculture described two traditional extra-large (> 60 g / 100 seeds) white seeded bean varieties, named "Gallega de Carballo" and "Planchada", which belong to the large great northern market class and are often cultivated by farmers in the Northwest of Spain. The selection made by antique farmers for large seeded types, together with natural selection in a very different environment of the original in the Americas, could be the origin of the extreme phenotype displayed by this group of varieties. Additionally, stochastic processes could be relevant in the early times when Columbus and other explorers collected seeds of different crops from American natives in the New World and when antique farmers harvest seeds from fields in Europe, since it is possible that many plants resulted in no seed production due to adaptive problems. Therefore the harvest, year after year, could no represent randomly the original variation coming from the Americas. An alternative explanation to the existence of these types of varieties could be occasional outcrossing among Mesoamerican and large-seeded Andean beans in farmer fields. Mesoamerican and Andean recombinant types An important feature is the existence of a high number of white seeded varieties which constitute intermediate forms or recombinants between the Mesoamerican and Andean genetic pools. The mixtures of seeds showing characteristic genetic patterns of the two common bean gene pools were described in common bean varieties from Italy (Piergiovanni et al., 2000) and from the Iberian Peninsula (Rodio et al., 2001, 2006) and they could be attributed to different selection made by farmers. The recombination could be explained because Iberian varieties, that are frequently a mixture of phenotypes with a common appearance, are often cultivated in small gardens in proximity, and under this circumstance

the exchange of seeds among neighbouring farms may result in gene flow. Thus, Iberian bean varieties could reflect the influence of both introduced germplasm, and also derived from secondary diversification. BREEDING ASSESSMENT Thus, Iberian common bean germplasm is more complex than was previously thought and contains additional diversity that remains to be explored for its practical value. Mesoamerican and Andean varieties, as well as the European new forms, could have disseminated to other parts of Europe, which, as mentioned above, would justify the widest genetic variation observed in European germplasm. The intergene pool recombinants overcome the typical association among traits. They also displayed new gene combinations that represent a novel genetic variation in the common bean genetic pools in Europe, giving a challenge for its use in plant breeding. The European recombinants and extreme bean types could constitute bridging germplasm for the transfer of useful genes between the two gene pools, if direct crosses cannot achieve this. The value of these unique recombinant and extreme varieties in common bean needs to be determined in order to create useful genetic variation for maximizing gains from selection, broadening the genetic base of commercial cultivars, and increase the durability of resistance to diseases (Singh, 2001). Higher levels and more stable resistance to a much broader range of pathogenic populations could be achieved as the different groups of pathogen populations have co-evolved with each common bean genetic pool. Thus, these intermediate forms would have important implications in genetic improvement. Recent studies also highlight the large percentage of bean varieties that are carrying alleles from both original American pools and the broad genetic diversity shown by the European common bean germplasm at molecular level (DNA) (Santalla et al., 2010). Increasing the knowledge about the variability of the European bean, particularly the Mediterranean genotypes, is essential in order to select the most suitable for breeding, both for hybridization and selection of lines within adapted varieties. As well as this, it is quite important to gain a better understanding about what part of the genome of bean varieties from the New World are still present in the current European varieties. The novel variation among the common European bean varieties here described could be the core of a new Old World gene pool arising after several centuries of evolution of the species outside the Americas.

ACKNOWLEDGEMENTS Research was supported by the AGF97-0324, RF95-008-C4-1, RF99-003-C5-1 and AGL2008-2091 projects from the Spanish Government and INCITE07PXI403088ES from the Galician Government. The authors are grateful to the National Center for Plant Genetic Resources (CRF-INIA, Alcal de Henares, Spain), to CIAT (Cali, Colombia) and Dr. M. West (WRPIS, Pullman, USA) for supplying bean germplasm, and to Diputacin of Pontevedra for farming facilities. REFERENCES DEBOUCK DG and SMARTT J, 1995. Bean, In: SMARTT J, SIMMONDS NW (eds.), Evolution of crop plants. 2nd edn. Longman Scientific and Technical, England, p.287296. GEPTS P and BLISS FA, 1988. Dissemination pathways of common bean (Phaseolus vulgaris, Fabaceae) deduced from phaseolin electrophoretic variability. II. Europe and Africa, Economic Botany n.42, p.86104. GEPTS P, OSBORN TC, RASHKA K and BLISS FA, 1986. Phasseolin-protein variability in wild forms and populations of the common bean (Phaseolus vulgaris): evidence for multiple centers of domestication, Economic Botany n.40, p.451-468. GIL J and DE RON AM, 1992. Variation in Phaseolus vulgaris in the Northwest of the Iberian Peninsula, Plant Breeding, n.109, p.313-319. HALEY SD, MIKLAS PH, AFANADOR L and KELLY JD, 1994. Random amplified polymorphic DNA (RAPD) marker variability between and within gene pools of common bean, Journal American Society Horticultural Science, n.119, p.122-125. LIOI L, 1989. Geographical variation of phaseolin patterns in an Old World collection of Phaseolus vulgaris, Seed Science Technology, n.17, p.317-324. NEGRI V and TOSTI N, 2002. Phaseolus genetic diversity maintained on-farm in central Italy, Genetic Resources Crop Evolution, n.47, p.489-495. ORTWIN-SAUER C, 1966. The early Spanish man. University of California Press, Berkeley and Los Angeles, p.51298. PIERGIOVANNI AR, CERBINO D and BRANDI M, 2000. The common bean populations from Basilicata (Sourthern Italy). An evaluation of their variation, Genetic Resources Crop Evolution, n.47, p.489-495. PUERTA-ROMERO J, 1961. Variedades de juda cultivadas en Espaa. Monografas INIA 11. Madrid. Spain. p.798 RODIO AP, SANTALLA M, MONTERO I, CASQUERO PA and DE RON AM, 2001. Diversity in common bean (Phaseolus vulgaris L.) germplasm from Portugal, Genetic Resources Crop Evolution, n.48, p.409-417. RODIO AP, GONZLEZ AM, SANTALLA M, DE RON AM and SINGH SP, 2006. Novel genetic variation in common bean from the Iberian Peninsula, Crop Science, n.46,

p.2540-2546. SANTALLA MA, DE RON AM and VOYSEST O, 2001. European bean market classes. In: AMURRIO JM, SANTALLA M and DE RON AM (eds.) Catalogue of Bean Genetic Resources. PHASELIEU FAIR 3463-MBG-CSIC. Fundacin Pedro Barri de la Maza. Spain. p.79-94. SANTALLA M, RODIO AP and DE RON AM, 2002. Allozyme evidence supporting Southwestern Europe as a secondary center of genetic diversity for the common bean, Theoretical Applied Genetics, n.104, p.934-944. SANTALLA M., DE RON AM and DE LA FUENTE M, 2010. Integration of genome and phenotype scanning gives evidence of genetic structure in Mesoamerican common bean (Phaseolus vulgaris L.) landraces from the southwest of Europe. Theorethical Applied Genetics, DOI:10.1007/s00122-010-1282-0. SINGH SP, 2001. Broadening the genetic base of common bean populations: a review, Crop Science n.41, p.1659-1675. SINGH SP, GEPTS P and DEBOUCK DG, 1991a. Races of common bean (Phaseolus vulgaris, Fabaceae) , Economic Botany, n.45, p.379-396. SINGH SP, NODARI, R and GEPTS P, 1991b. Genetic diversity in cultivated common bean. I. Allozymes, Crop Science, n.31, p.19-23. ZEVEN AC, 1997. The introduction of the common bean (Phaseolus vulgaris L.) into Western Europe and the phenotypic variation of dry beans collected in the Netherlands in 1946, Euphytica, n.94, p.319-328.

Table 1. Description of Mesoamerican and Andean typical bean varieties, Mesoamerican extreme, Mesoamerican recombinant, and Andean recombinant types (PHA varieties code at the MBG-CSIC).
GENOTYPE VARIETIES Phaseolin (*) M (*) PHA-0151 Mesoamerican typical M Skdh-103, Diap195 , Me-100, Rbc100 , Mdh1-100, Mdh2-100 Skdh-100, Diap1-100, Me100 , Rbc-98, Mdh1-100, Mdh2-100 Skdh-103, Diap195 , Me-100, Rbc100 , Mdh1-100, Mdh2-100 Diap1-95, Me-98 Skdh-100, Rbc98 , Mdh1-100, Mdh2-100 29 Isozymes A (*) PHENOTYPE Seed size (g/100 seeds) M (*) 37 A(*) Great northern Market class

PHA-0400 Andean typical

47

White kidney

PHA-0419 Mesoamerican extreme

75

Large great northern

PHA-0430 Mesoamerican recombinant

Small white

PHA-0589 Mesoamerican recombinant

Diap1-95, Rbc-100 Skdh-100, Me100, Mdh1-100, Mdh2-100

44

Navy

PHA-0593 Mesoamerican recombinant PHA-0481 Andean recombinant

Skdh-103, Rbc-102 Diap1-100, Me100 , Mdh1-100, Mdh2-100 Diap1-95, Me-98, Skdh-100, Rbc-100/102 Mdh1-100, Mdh2-100 Skdh-103, Rbc-100 Skdh100, Diap1-100, Me100 , Mdh1-100, Mdh2-100 Skdh-103, Rbc-100 Diap1-100, Me100 , Mdh1-100, Mdh2-100 33

45

Hook

Guernikesa

PHA-0501 Andean recombinant

46

Negro brillante

PHA-0554 Andean recombinant

50

Canellini

PHA-0917 Andean recombinant

Diap1-95, Rbc-100 Skdh-100, Me100 , Mdh1-100, Mdh2-100

102

Favada

(*) M: Mesoamerican, A: Andean