Abstract
Anthracnose, caused by Colletotrichum trifolii, is a severe disease of alfalfa (Medicago sativa). The RCT1 gene, isolated from the model legume M. truncatula, is a candidate gene to explain genetic variation for anthracnose resistance in alfalfa. A bulk segregant analysis was carried out to test this hypothesis: from each of eight alfalfa varieties, 15 resistant (R) plants and 15 susceptible (S) plants were selected and DNA was extracted. The whole gene including the upstream and downstream regions (a total of 14 kb) was amplified by PCR for each individual and the R and S plants were pooled for each variety. Sequencing was carried out using the next generation sequencer 454 (Roche). The sequence reads, that averaged 295 bp, were assembled to produce consensus sequences that can be considered as alleles. Considering the five exons of the gene, five regions contained clear deletion/insertion polymorphism but these polymorphisms were present in both the R and S pools. Individual genotyping for these indels indicated that different alleles were present but no specific allele was associated with the phenotype. These polymorphic regions in RCT1 seemed not to explain the variation of anthracnose resistance in alfalfa. However, the presence of one rare allele inducing a lack of function was associated with the resistance. A divergent selection for this allele would test its interest in breeding programs.
Keywords
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Ameline-Torregrosa C, Cazaux M, Danesh D, Chardon F, Cannon SB, Esquerre-Tugaye MT, Dumas B, Young ND, Samac DA, Huguet T, Jacquet C (2008) Genetic dissection of resistance to anthracnose and powdery mildew in Medicago truncatula. MPMI 21:61–69
Ariss JJ, Vandemark GJ (2007) Assessment of genetic diversity among nondormant and semidormant alfalfa populations using sequence-related amplified polymorphisms. Crop Sci 47:2274–2284
Barnes DK, Ostazeski SA, Schillinger JA, Hanson CH (1969) Effect of anthracnose (Colletotrichum trifolii) infection on yield, stand, and vigor of alfalfa. Crop Sci 9:344–346
Cazaux M (2008) Etude de la résistance de la légumineuse modèle Medicago truncatula à Colletotrichum trifolii, agent de l’anthracnose. PhD Thesis Université de Toulouse
Elgin JH Jr, Ostazeski SA (1985) Inheritance of resistance to race 1 and race 2 anthracnose in Arc and Saranac AR alfalfa. Crop Sci 25:861–865
Gondran J (1984) La verticilliose de la luzerne: détermination de l’agent causal, biologie du parasite, répartition géographique, dégâts et méthodes de lutte. Thèse Université U.E.R. Sciences de Poitiers
Herrmann D, Barre P, Santoni S, Julier B (2010) Association of a CONSTANS-LIKE gene to flowering and height in autotetraploid alfalfa. Theor Appl Genet 121:865–876
Julier B, Meusnier I (2010) Alfalfa breeding benefits from genomics of Medicago truncatula. Ratar Povrt/Field Veg Crop Res 47:395–402
Julier B, Bournoville R, Landré B, Ecalle C, Carré S (2004) Genetic analysis of lucerne (Medicago sativa L.) seedling resistance to pea aphid (Acyrtosiphon pisum Harris). Euphytica 138:133–139
Mackie JM, Musial JM, O’Neill NR, Irwin JAG (2003) Pathogenic specialisation within Colletotrichum trifolii in Australia, and lucerne cultivar reactions to all known Australian pathotypes. Aust J Agric Res 54:829–836
Mackie JM, Musial JM, Armour DJ, Phan HTT, Ellwood SE, Aitken KS, Irwin JAG (2007) Identification of QTL for reaction to three races of Colletotrichum trifolii and further analysis of inheritance of resistance in autotetraploid lucerne. Theor Appl Genet 114:1417–1426
Michelmore RW, Paran I, Kesseli RV (1991) Identification of markers linked to disease-resistance genes by bulked segregant analysis: a rapid method to detect markers in specific genomic regions by using segregating populations. Proc Natl Acad Sci 88:9828–9832
O’Neill N (1991) Anthracnose resistance. In: CC Fox et al. (ed) Standard tests to characterise alfalfa cultivars, D1. North American alfalfa improvement conference
Pierre JB, Bogard M, Herrmann D, Huyghe C, Julier B (2011) A CONSTANS-like gene candidate that could explain most of the genetic variation for flowering date in Medicago truncatula. Mol Breed 28:25–35
Raynal G, Gondran J, Bournoville R, Courtillot M (1989) Ennemis et maladies des prairies. INRA, Paris
Yang S, Gao M, Deshpande S, Lin S, Roe BA, Zhu H (2007) Genetic and physical localization of an anthracnose resistance gene in Medicago truncatula. Theor Appl Genet 116:45–52
Yang S, Gao M, Xu C, Gao J, Deshpande S, Lin S, Roe BA, Zhu H (2008) Alfalfa benefits from Medicago truncatula: the RCT1 gene from M. truncatula confers broad-spectrum resistance to anthracnose in alfalfa. Proc Natl Acad Sci 105:12164–12169
Acknowledgements
We thank the French Ministry of Agriculture for financial support (Contrat de Branches 2008–11) and ACVF for scientific and technical contribution. We thank J. Lluch and O. Bouchez from “Plateforme Génomique” of Toulouse (France) for 454 sequencing and S. Fouilloux from BioGeves at Surgères (France) for DNA quantification.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media Dordrecht
About this paper
Cite this paper
Julier, B., Meusnier, I., Alaux, L., Flajoulot, S., Barre, P., Gouzy, J. (2013). Role of the RCT1 Gene in Anthracnose Resistance in Alfalfa. In: Barth, S., Milbourne, D. (eds) Breeding strategies for sustainable forage and turf grass improvement. Springer, Dordrecht. https://doi.org/10.1007/978-94-007-4555-1_25
Download citation
DOI: https://doi.org/10.1007/978-94-007-4555-1_25
Published:
Publisher Name: Springer, Dordrecht
Print ISBN: 978-94-007-4554-4
Online ISBN: 978-94-007-4555-1
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)