Free Standard AU & NZ Shipping For All Book Orders Over $80!
Register      Login
Crop and Pasture Science Crop and Pasture Science Society
Plant sciences, sustainable farming systems and food quality
RESEARCH ARTICLE

A genetic linkage map in autotetraploid lucerne adapted to northern Australia, and use of the map to identify DNA markers linked to resistance to Phytophthora medicaginis

J. M. Musial A , K. S. Aitken B , J. M. Mackie A and J. A. G. Irwin A C
+ Author Affiliations
- Author Affiliations

A Cooperative Research Centre for Tropical Plant Protection, The University of Queensland, Brisbane, Qld 4072, Australia.

B CSIRO Plant Industry, Queensland Bioscience Precinct, St Lucia, Qld 4067, Australia.

C Corresponding author. Email: j.irwin@tpp.uq.edu.au

Australian Journal of Agricultural Research 56(4) 333-344 https://doi.org/10.1071/AR04317
Submitted: 20 December 2004  Accepted: 1 March 2005   Published: 26 April 2005

Abstract

Phytophthora root rot, caused by Phytophthora medicaginis, is a major limitation to lucerne (Medicago sativa L.) production in Australia and North America. Quantitative trait loci (QTLs) involved in resistance to P. medicaginis were identified in a lucerne backcross population of 120 individuals. A genetic linkage map was constructed for tetraploid lucerne using 50 RAPD (randomly amplified polymorphic DNA), 104 AFLP (amplified fragment length polymorphism) markers, and one SSR (simple sequence repeat or microsatellite) marker, which originated from the resistant parent (W116); 13 markers remain unlinked. The linkage map contains 18 linkage groups covering 2136.5 cM, with an average distance of 15.0 cM between markers. Four of the linkage groups contained only either 2 or 3 markers. Using duplex markers and repulsion phase linkages the map condensed to 7 homology groups and 2 unassigned linkage groups. Three regions located on linkage groups 2, 14, and 18, were identified as associated with root reaction and the QTLs explained 6–15% of the phenotypic variation. The research also indicates that different resistance QTLs are involved in conferring resistance in different organs. Two QTLs were identified as associated with disease resistance expressed after inoculation of detached leaves. The marker, W11-2 on group 18, identified as associated with root reaction, contributed 7% of the phenotypic variation in leaf response in our population. This marker appears to be linked to a QTL encoding a resistance factor contributing to both root and leaf reaction. One other QTL, not identified as associated with root reaction, was positioned on group 1 and contributed to 6% of the variation. This genetic linkage map provides an entry point for future molecular-based improvement of lucerne in Australia, and markers linked to the QTLs we have reported should be useful for marker-assisted selection for partial resistance to P. medicaginis in lucerne.

Additional keywords: alfalfa, SDRF, disease resistance, molecular markers, QTL mapping.


Acknowledgments

The authors thank the CRC for Tropical Plant Protection and the GRDC (Grains Research and Development Corporation) for providing funding support for the project. We also thank Dr C. L. McIntyre, Prof. E. T. Bingham, Prof. T. C. Osborn, Prof. M. J. Havey, and Dr B. Julier for discussions on autotetraploid genetics.


References


Baquerizo E, Desplanque B, Santoni S (2001) Isolation and characterization of microsatellites in Medicago truncatula (Leguminosae). Molecular Ecology Notes 1, 1–3. open url image1

Barcaccia G, Albertini E, Tavoletti S, Falcinelli M, Veronesi F (1999) AFLP fingerprinting in Medicago spp. Its development and application in linkage mapping. Plant Breeding 118, 335–340.
Crossref | GoogleScholarGoogle Scholar | open url image1

Brouwer DJ, Duke SH, Osborn TC (2000) Mapping genetic factors associated with winter hardiness, fall growth and freezing injury in autotetraploid alfalfa. Crop Science 40, 1387–1396. open url image1

Brouwer DJ, Osborn TC (1999) A molecular marker linkage map of tetraploid alfalfa (Medicago sativa L.). Theoretical and Applied Genetics 99, 1194–1200.
Crossref | GoogleScholarGoogle Scholar | open url image1

Brummer EC, Bouton JH, Kochert G (1993) Development of an RFLP map in diploid alfalfa. Theoretical and Applied Genetics 86, 329–332.
Crossref | GoogleScholarGoogle Scholar | open url image1

Choi H-K, Kim D, Uhm T, Limpens E, Lim H , et al . (2004) A sequence-based genetic map of Medicago truncatula and comparison of marker colinearity with M. sativa.  Genetics 166, 1463–1502.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Clements RJ, Turner JW, Irwin JAG, Langdon PW, Bray RA (1984) Breeding disease resistant, aphid resistant lucerne for sub tropical Queensland. Australian Journal of Experimental Agriculture and Animal Husbandry 24, 178–188.
Crossref | GoogleScholarGoogle Scholar | open url image1

Cordero J, Skinner D (2002) Isolation from alfalfa of resistance gene analogues containing nucleotide binding sites. Theoretical and Applied Genetics 104, 1283–1289.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Molecular Biology Reporter 1, 19–21. open url image1

Diwan N, Bhagwat AA, Bauchan GR, Cregan PB (1997) Simple sequence repeat (SSR) DNA markers in alfalfa and perennial and annual Medicago species. Genome 40, 887–895. open url image1

Echt CS, Kidwell KK, Knapp SJ, Osborn TC, McCoy TJ (1994) Linkage mapping in diploid alfalfa (Medicago sativa). Genome 37, 61–71.
PubMed |
open url image1

Erwin DC (1966) Varietal reaction of alfalfa to Phytophthora megasperma and variation in virulence of the causal fungus. Phytopathology 56, 653–657. open url image1

Fisher RA (1947) The theory of linkage in polysomic inheritance. Philosophical Transactions of the Royal Society of Britain 233, 55–87. open url image1

Havey MJ, Maxwell DP (1987) Inheritance of Phytophthora root rot resistance in two diploid alfalfa species. Crop Science 27, 225–228. open url image1

Havey MJ, Maxwell DP, Irwin JAG (1987) Independent inheritance of genes conditioning resistance to Phytophthora root rot from diploid and tetraploid alfalfa. Crop Science 27, 873–879. open url image1

Hill MJ (1996) Potential adaptation zones for temperate pasture species as constrained by climate: a knowledge-based logical modelling approach. Australian Journal of Agricultural Research 47, 1095–1117.
Crossref | GoogleScholarGoogle Scholar | open url image1

Humphry M, Konduri V, Lambrides CJ, Magner T, McIntyre CL, Aitken EAB, Liu CJ (2002) Development of a mungbean (Vigna radiata) RFLP linkage map and its comparison with lablab (Lablab purpureus) reveals a high level of colinearity between the two genomes. Theoretical and Applied Genetics 105, 160–166.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Irwin JAG (1974) Reaction of lucerne cultivars to Phytophthora megasperma, the cause of root rot in Queensland. Australian Journal of Experimental Agriculture and Animal Husbandry 14, 561–565.
Crossref | GoogleScholarGoogle Scholar | open url image1

Irwin JAG (1977) Factors contributing to poor resistance of lucerne in southern Queensland. Australian Journal of Experimental Agriculture and Animal Husbandry 17, 998–1003.
Crossref | GoogleScholarGoogle Scholar | open url image1

Irwin JAG, Lloyd DL, Lowe KF (2001) Lucerne biology and genetic improvement — an analysis of past activities and future goals in Australia. Australian Journal of Agricultural Research 52, 699–712.
Crossref | GoogleScholarGoogle Scholar | open url image1

Irwin JAG, Maxwell DP, Bingham ET (1981a) Inheritance of resistance to Phytophthora megasperma in diploid alfalfa. Crop Science 21, 271–276. open url image1

Irwin JAG, Maxwell DP, Bingham ET (1981b) Inheritance of resistance to Phytophthora megasperma in tetraploid alfalfa. Crop Science 21, 277–283. open url image1

Irwin JAG, Miller SA, Maxwell DP (1979) Alfalfa seedling resistance to Phytophthora megasperma.  Phytopathology 69, 1051–1055. open url image1

Irwin JAG, Musial JM, Mackie JM, Basford KE (2003) Utility of cotyledon and detached leaf assays for assessing root reactions of lucerne to Phytophthora root rot caused by Phytophthora medicaginis.  Australasian Plant Pathology 32, 263–268.
Crossref | GoogleScholarGoogle Scholar | open url image1

Julier B, Fajoulot S, Barre P, Cardinet G, Santoni S, Huguet T, Huyghe C (2003) Construction of two genetic linkage maps in cultivated tetraploid alfalfa (Medicago sativa) using microsatellite and AFLP markers. BMC Plant Biology 3, 9.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Kaló P, Endre G, Zimányi L, Csanádi G, Kiss GB (2000) Construction of an improved linkage map of diploid alfalfa (Medicago sativa). Theoretical and Applied Genetics 100, 641–657.
Crossref | GoogleScholarGoogle Scholar | open url image1

Kidwell KK, Woodfield DR, Bingham ET, Osborn TC (1994) Molecular marker diversity and yield of isogenic 2x and 4x single-crosses of alfalfa. Crop Science 34, 784–788. open url image1

Kiss GB, Csanádi G, Kálmán K, Kaló P, Okrész L (1993) Construction of a basic genetic map for alfalfa using RFLP, RAPD, isozyme and morphological markers. Molecular and General Genetics 238, 129–137.
PubMed |
open url image1

Lander ES, Green P, Abrahamson J, Barlow A, Daly MJ, Lincoln SE, Newburg L (1987) MapMaker: an interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics 1, 174–181.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Lu NSJ, Barnes DK, Frosheiser FI (1973) Inheritance of Phytophthora root rot resistance in alfalfa. Crop Science 13, 714–717. open url image1

Manglitz GR, Ratcliffe RH (1988) Insects and mites. ‘Alfalfa and alfalfa improvement’. (Ed. AA Hanson) pp. 671–704. (American Society of Agronomy: Madison, WI)

Manly KF, Cudmore RH, Meer JM (2001) Map Manager QTX, cross-platform software for genetic mapping. Mammalian Genome 12, 930–932.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Mather K (1936) Segregation and linkage in autotetraploids. Journal of Genetics 32, 287–314. open url image1

Mather, K (1951). ‘The measurement of linkage in heredity.’ (Methuen and Co. Ltd: London)

Michaud R, Lehman WF, Rumbaugh MD (1988) World distribution and historical development. ‘Alfalfa and alfalfa improvement. Agronomy Monograph 29’. pp. 25–91. (American Society of Agronomy: Madison, WI)

Musial JM, Basford KE, Irwin JAG (2002) Analysis of genetic diversity within Australian lucerne cultivars and implications for future genetic improvement. Australian Journal of Agricultural Research 53, 629–636.
Crossref | GoogleScholarGoogle Scholar | open url image1

Obert DE, Skinner DZ, Stuteville DL (2000) Association of AFLP markers with downy mildew resistance in autotetraploid alfalfa. Molecular Breeding 6, 287–294.
Crossref | GoogleScholarGoogle Scholar | open url image1

Oram, RN (1990). ‘Register of Australian herbage cultivars.’ 3rd edn . (CSIRO: Melbourne, Vic.)

Pearson CJ, Brown R, Collins WJ, Archer KA, Wood MS, Petersen C, Boothe B (1997) An Australian temperate pastures database. Australian Journal of Agricultural Research 48, 453–465.
Crossref | GoogleScholarGoogle Scholar | open url image1

da Silva JAG, Honeycutt RJ, Burnquist W, Al-Janabi SM, Tanksley SD, Sobral BWS (1995) Saccharum spontaneum L. ‘SES 208’ genetic linkage map combining RFLP and PCR-based markers. Molecular Breeding 1, 165–179.
Crossref | GoogleScholarGoogle Scholar | open url image1

Stanford EH (1951) Tetrasomic inheritance in alfalfa. Agronomy Journal 43, 222–225. open url image1

Stuteville, DL ,  and  Erwin, DC (1990). ‘Compendium of alfalfa diseases.’ 2nd edn . (APS Press: St Paul, MN)

Tanksley SD, Young ND, Paterson AH, Bonierbale MW (1989) RFLP mapping in plant breeding: new tools for an old science. Bio/Technology 7, 257–264.
Crossref | GoogleScholarGoogle Scholar | open url image1

Tavoletti S, Veronesi F, Osborn TC (1996) RFLP linkage map of an alfalfa meiotic mutant based on a F1 population. Journal of Heredity 87, 167–170. open url image1

Thies JA, Barnes DK (1991) Phytophthora root rot resistance. ‘Standard tests to characterize alfalfa cultivars’. (Eds CC Fox, R Berberet, FA Gray, CR Grau, DL Jessen, MA Peterson) p. D8. (North American Alfalfa Improvement Conference)


Thoquet P, Gherardi M, Journet EP, Kereszt A, Ane JM, Prosperi JM, Huguet T (2002) The molecular genetic linkage map of the model legume Medicago truncatula: an essential tool for comparative legume genomics and the isolation of agronomically important genes. BMC Plant Biology 2, 1.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Tysdal HM, Kiesselbach TA, Westover HL (1942) Alfalfa breeding. Nebraska Agricultural Experiment Station Research Bulletin 124.

Vos P, Hogers R, Bleeker M, Reijans M, van de Lee T , et al . (1995) AFLP: a new technique for DNA fingerprinting. Nucleic Acids Research 23, 4407–4414.
PubMed |
open url image1

Zhao J, Meng J (2003) Genetic analysis of loci associated with partial resistance to Sclerotinia sclerotiorum in rapeseed (Brassica napus L.). Theoretical and Applied Genetics 106, 759–764.
PubMed |
open url image1

Zhu H, Cannon SB, Young ND, Cook DR (2002) Phylogeny and genomic organization of the TIR and Non-TIR NBS-LRR resistance gene family in Medicago truncatula.  Molecular Plant-Microbe Interactions 15, 529–539.
PubMed |
open url image1