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Extension of the linkage map in Citrus using random amplified polymorphic DNA (RAPD) markers and RFLP mapping of cold-acclimation-responsive loci

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Genetic mapping with RAPD markers has been initiated in Citrus. Reproducible polymorphism of amplified DNA fragments was obtained with approximately half of the 140 random primers tested, revealing 266 segregating loci. These were tested for linkage using 60 BC1 progeny from an intergeneric cross of Citrus grandis (L.) Osb. x [Citrus grandis (L.) Osb. x Poncirus trifoliata (L.) Raf.]. A core linkage map was constructed that consists of nine linkage groups containing 109 RAPD markers and 51 previously-mapped RFLP and isozyme markers. A further 79 markers that could not be ordered unambiguously because of their allelic constitution were associated with individual linkage groups and are shown in relation to the core map. The core map has a total length of 1192 cM with an average distance of 7.5 cM between loci and is estimated to cover 70–80% of the genome. Loci with distorted segregation patterns clustered on several linkage groups. Individual clusters of loci were skewed in allelic composition toward one or the other parent, usually C. grandis. This relatively-saturated linkage map will eventually be used to identify quantitative trait loci for cold and salt-tolerance in Citrus. As a beginning we have mapped three loci detected by a cold-acclimation-responsive cDNA.

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  1. Ausubel FM, Rrent R, Kinston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1989) Current protocols in molecular biology. Greene Publ Assoc, Wiley-Interscience, New York

  2. Durham RE, Liou PC, Gmitter FG, Jr, Moore GA (1992) Linkage of restriction fragment length polymorphisms and isozymes in Citrus. Theor Appl Genet 84:39–48

  3. Gebhardt C, Ritter E, Debener T, Schachtschabel V, Wakemeier B, Uhrig H, Salamini F (1989) RFLP analysis and linkage mapping in Solanum tuberosum. Theor Appl Genet 78:65–75

  4. Guerra MS (1984) Cytogenetics of Rutaceae. II. Nuclear DNA content. Caryologia 37:219–226

  5. Helentjaris T, Slomcum M, Wright S, Schaefer A, Nienhuis J (1986) Construction of genetic linkage maps in maize and tomato using restriction fragment length polymorphisms. Theor Appl Genet 72:761–769

  6. Jarrell DC, Roose ML, Traugh SN, Kupper RS (1992) A genetic map of citrus based on the segregation of isozymes and RFLPs in an intergeneric cross. Theor Appl Genet 84:49–56

  7. Klein-Lankhorst RM, Vermunt A, Weide R, Liharska T, Zabel P (1991) Isolation of molecular markers for tomato (L. esculentum) using random amplified polymorphic DNA (RAPD). Theor Appl Genet 83:108–114

  8. Kosambi DD (1944) The estimation of map distances from recombination values. Ann Eugen 12:172–175

  9. 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

  10. Landry BS, Kesseli RV, Farrara B, Michelmore RW (1987) A genetic map of lettuce (Lactuca sativa L.) with restriction fragment length polymorphism, isozyme, disease resistance and morphological markers. Genetics 16:331–337

  11. Landry BS, Hubert N, Crete R, Chang MS, Etoh T (1992) A genetic map for Brassica oleracea based on RFLP markers detected with expressed DNA sequences and mapping of resistance genes of race 2 of Plasmodiophora brassicae (Woronin). Genome 35:409–420

  12. Leyser HMO, Lincoln CA, Timpte C, Lammer D, Turner J, Estelle M (1993) Arabidopsis auxin-resistance gene AXR1 encodes a protein related to ubquitin-activating enzyme E1. Nature 364:161–164

  13. Lincoln S, Daly M, Lander E (1992) Constructing genetic maps with MAPMAKER/EXP 3.0. Whitehead Institute Technical Report, 3rd ed.

  14. Liou PC (1990) A molecular study of the Citrus genome through restriction fragment length polymorphism and isozyme analysis. Doctoral Dissertation, University of Florida, Gainesville, Florida

  15. Martin GB, Brommonschenckel SH, Chunwongse J, Frary A, Ganal, MW, Spivey R, Wu T, Earle ED, Tanksley SD (1993) Map-based cloning of a protein kinase gene conferring disease resistance in tomato. Science 262:1432–1436

  16. Michelmore RW, Paran I, Kesseli V (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 USA 88:9828–9832

  17. Monroy AF, Castonguay Y, Laberge S, Sarhan F, Vezina LP, Dhindsa RS (1993) A new cold-induced alfalfa gene is associated with enhanced hardening at subzero temperature. Plant Physiol 102:873–879

  18. Pillen K, Steinrücken G, Wricke G, Herrmann RG, Jung C (1992) A linkage map of sugar beet (Beta vulgaris L.). Theor Appl Genet 84:129–135

  19. Prince J, Pochard E, Tanksley SD (1993) Construction of a molecular linkage map of pepper and a comparison of synteny with tomato. Genome 36:404–417

  20. Reiter RS, Williams J, Feldmann KA, Rafalski JA, Tinger SV, Scolnik PA (1992) Global and local genome mapping in Arabidopsis thaliana by using recombinant inbred lines and random amplified polymorphic DNAs. Proc Natl Acad Sci USA 89:1477–1481

  21. Soost RK, Cameron JW (1975) Citrus. In: Janick J, Moore JN (eds) Advances in fruit breeding. Purdue University Press, West Lafayette, Indiana, pp 507–540

  22. Suiter K, Wendel J, Case J (1983) LINKAGE-1: a PASCAL computer program for the detection and analysis of genetic linkage. J Hered 74:203–204

  23. Stam P (1993) Construction of integrated genetic linkage maps by means of a new computer package: JoinMap. Plant J 3:739–744

  24. Tanksley SD, Young ND, Paterson AH, Bonierbale MW (1989) RFLP mapping in plant breeding: new tools for an old science. Biotechnology 7:257–264

  25. Torres AM, Mau-Lostovicka T, William TE, Soost PK (1985) Segregation distortion and linkage of Citrus and Poncirus isozyme genes. J Hered 76:289–294

  26. Torres AM, Weeden NF, Martin A (1993) Linkage among isozyme, RFLP and RAPD markers in Vicia faba. Theor Appl Genet 85:937–945

  27. Tulsieram LK, Glaubitz JC, Kiss G, Carlson JE (1992) Single-tree genetic linkage mapping in conifers using haploid DNA from megagametophytes. Biotechnology 10:686–690

  28. Weeden NF, Timmerman GM, Hemmat M, Kneen BE, Lodhi MA (1992) Inheritance and reliability of RAPD markers. Application of RAPD technology to plant breeding: joint plant breeding symposia series. Minneapolis, Minnesota

  29. Welsh J, McClelland M (1990) Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res 18:7213–7218

  30. Williams JGK, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research 18:6531–6535

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Communicated by F. Salamini

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Cai, Q., Guy, C.L. & Moore, G.A. Extension of the linkage map in Citrus using random amplified polymorphic DNA (RAPD) markers and RFLP mapping of cold-acclimation-responsive loci. Theoret. Appl. Genetics 89, 606–614 (1994).

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  • Genetic map
  • Molecular markers
  • Fruit breeding