Abstract
We have cloned and sequenced six RAPD fragments tightly linked to the Tm-1 gene which confers tomato mosaic virus (ToMV) resistance in tomato. The terminal ten bases in each of these clones exactly matched the sequence of the primer for amplifying the corresponding RAPD marker, except for one in which the 5′-endmost two nucleotides were different from those of the primer. These RAPD clones did not cross-hybridize with each other, suggesting that they were derived from different loci. From Southern-hybridization experiments, five out of the six RAPD clones were estimated to be derived from middle- or high-repetitive sequences, but not from any parts of the ribosomal RNA genes (rDNA), which are known to be tightly linked with the Tm-1 locus. The remaining clone appeared to be derived from a DNA family consisting of a few copies. These six RAPD fragments were converted to sequence characterized amplified region (SCAR) markers, each of which was detectable using a pair of primers having the same sequence as that at either end of the corresponding RAPD clone. All pairs of SCAR primers amplified distinct single bands whose sizes were the same as those of the RAPD clones. In four cases, the SCAR markers were present in the line with Tm-1 but absent in the line without it, as were the corresponding RAPD markers. In the two other cases, the products of the same size were amplified in both lines. When these SCAR products were digested with different restriction endonucleases which recognize 4-bp sequences, however, polymorphisms in fragment length were found between the two lines. These co-dominant markers are useful for differentiating heterozygotes from both types of homozygote.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Adam-Blondon AF, Sévignac M, Bannerot H, Dron M (1994) SCAR, RAPD and RFLP markers linked to a dominant gene (Are) conferring resistance to anthracnose in common bean. Theor Appl Genet 88:865–870
Delourme R, Bouchereau A, Hubert N, Renard M (1994) Identification of RAPD markers linked to a fertility restorer gene for the Ogura radish cytoplasmic male sterility of rapeseed (Brassica napus L.). Theor Appl Genet 88:741–748
Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 12:13–15
Foolad MR, Jones RA, Rodriguez RL (1993) RAPD markers for constructing intraspecific tomato genetic maps. Plant Cell Rep 12:293–297
Gerlach WL, Bedbrook JR (1979) Cloning and characterization of ribosomal RNA genes from wheat and barley. Nucleic Acids Res 7:1869–1885
Holmes FO (1957) True-breeding resistance in tomato to infection by tobacco-mosaic virus. Phytopathology 47:16–17
Kreike CM, de Koning JRA, Krens FA (1990) Non-radioactive detection of single-copy DNA-DNA hybrids. Plant Mol Biol Rep 8:172–179
Levesque H, Vedel F, Mathieu C, de Courcel AGL (1990) Identification of a short rDNA spacer sequence highly specific of a tomato line containing Tm-1 gene introgressed from Lycopersicon hirsutum. Theor Appl Genet 80:602–608
Maisonneuve B, Bellec Y, Anderson P, Michelmore RW (1994) Rapid mapping of two genes for resistance to downy mildew from Lactuca serriola to existing clusters of resistance genes. Theor Appl Genet 89:96–104
Meshi T, Motoyoshi F, Adachi A, Watanabe Y, Takamatu N, Okada Y (1988) Two concomitant base substitutions in the putative replicase genes of tobacco mosaic virus confer the ability to overcome the effects of a tomato resistance gene, Tm-1. EMBO J 7:1575–1581
Motoyoshi F, Oshima N (1975) Infection with tobacco mosaic virus of leaf mesophyll protoplasts from susceptible and resistant lines of tomato. J Gen Virol 29:81–91
Motoyoshi F, Oshima N (1977) Expression of genetically controlled resistance to tobacco mosaic virus infection in isolated tomato leaf mesophyll protoplasts. J Gen Virol 34:499–506
Ohmori T, Murata M, Motoyoshi F (1995a) Identification of RAPD markers linked to the Tm-2 locus in tomato. Theor Appl Genet 90:307–311
Ohmori T, Murata M, Motoyoshi F (1995b) RAPD markers linked to the tomato mosaic virus resistance gene, Tm-1, in tomato. Jpn J Genet 70:179–184
Paran I, Michelmore RW (1993) Development of reliable PCR-based markers linked to downy mildew resistance genes in lettuce. Theor Appl Genet 85:985–993
Reiter RS, Williams JGK, Feifmann KA, Rafalski JA, Tingey 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
Sambrook L, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold spring Harbor Laboratory Press, Cold Spring Harbor, New York
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Smith JWM, Ritchie DB (1983) A collection of near-isogenic lines of tomato: research tool of the future? Plant Mol Biol Rep 1:41–45
Tanksley SD, Ganal MW, Prince JP, de Vicente MC, Bonierbale MW, Broun P, Fulton TM, Giovannoni JJ, Grandillo S, Martin GB, Messeguer R, Miller JC, Miller L, Paterson AH, Pineda O, Röder MS, Wing RA, Wu W, Young ND (1992) High-density molecular linkgae maps of the tomato and potato genomes. Genetics 132:1141–1160
Watanabe Y, Kishibayashi N, Motoyoshi F, Okada Y (1987) Characterization of Tm-1 gene action on replication of common isolates and a resistance-breaking isolate of TMV. Virology 161:527–532
Welsh J, McClelland M (1990) Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res 24:7213–7218
Williams JGK, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18:6531–6535
Author information
Authors and Affiliations
Additional information
Communicated by M. Koornneef
Rights and permissions
About this article
Cite this article
Ohmori, T., Murata, M. & Motoyoshi, F. Molecular characterization of RAPD and SCAR markers linked to the Tm-1 locus in tomato. Theoret. Appl. Genetics 92, 151–156 (1996). https://doi.org/10.1007/BF00223369
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00223369