Summary
Alfalfa (Medicago sativa L.) is a major forage crop throughout the world. Although alfalfa has many desirable traits, continued breeding is required to incorporate pest resistances and other traits. We conducted this study to determine the amount of restriction fragment length polymorphism (RFLP) variability present within and between diploid and tetraploid alfalfa populations, and whether or not this variability is sufficient for construction of an RFLP map. Diploid plants from M. sativa ssp. falcata, ssp. coerulea, and ssp. sativa and tetraploid spp. sativa cultivars ‘Apollo,’ ‘Florida 77,’ and ‘Spredor 2’ were included. A total of 19 cDNA clones was probed onto genomic Southern blots containing DNA digested by EcoRI, HindIII, or BamHI. Phylogenetic trees were produced, based on parsimony analysis of shared restriction fragments. Evidence for extensive gene duplication was found; most probes detected complex patterns of restriction fragments. Large amounts of variation are present within all diploid subspecies. M. sativa ssp. falcata plants formed clusters distinct from ssp. sativa or ssp. coerulea plants, which were not distinctly clustered. Some M. sativa ssp. falcata plants were more similar to the other groups than to other plants within ssp. falcata. Variation among tetraploid cultivars showed that Florida 77 and Apollo had more similarities than either showed with Spredor 2. All three cultivars showed large within-population variation, with Apollo being the most diverse and Spredor 2 the least. Based on these results, development of an RFLP map at the diploid level appears possible. Also, differentiation of cultivars, particularly ones of divergent origin, seems possible based on RFLP patterns.
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References
Anonymous (1980) Variety publication no. LXXXIII. Assn. of Official Seed Certifying Agencies
Barnes DK, Golpen BP, Baylor JE (1988) Highlights in the USA and Canada. In: Hanson AA, Barnes DK, Hill RR (eds) Alfalfa and alfalfa improvement. ASA, Madison/WI, pp 1–24
Bernatzky R, Tanksley SD (1986a) Majority of random cDNA clones correspond to single loci in the tomato genome. Mol Gen Genet 203: 8–14
Bernatzky T, Tanksley SD (1986b) Toward a saturated linkage map in tomato based on isozymes and random cDNA sequences. Genetics 112:887–898
Bingham ET (1991) Registration of isogenic populations of diploid and tetraploid alfalfa, W2xiso-1 and W4xiso-1. Crop Sci 31:496
Bingham ET, McCoy TJ (1979) Cultivated alfalfa at the diploid level: origin, reproductive stability, and yield of seed and forage. Crop Sci 19:97–100
Brummer EC, Bouton JH (dy1991) Plant traits associated with grazing-tolerant alfalfa. Agron J 83:Nov–Dec
Dallas JF (1988) Detection of DNA “fingerprints” of cultivated rice by hybridization with a human minisatellite DNA probe. PNAS 85:6831–6835
Dunbier MW, Bingham ET (1975) Maximum heterozygosity in alfalfa: results using haploid-derived autotetraploids. Crop Sci 15:527–531
Feinberg AP, Vogelstein B (1984) A technique for radiolabelling DNA restriction fragments to a high specific activity. Anal Biochem 132:6–13
Figdore SS, Kennard WC, Song KM, Slocum MK, Osborn TC (1988) Assessment of the degree of restriction fragment length polymorphism in Brassica. Theor Appl Genet 75:833–840
Helentjaris T, Slocum 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
Horner ES (1970) Registration of Florida 66 alfalfa. Crop Sci 10:456
Horner ES, Ruelke OC (1981) Registration of Florida 77 alfalfa. Crop Sci 21:797
Keim P, Shoemaker RC, Palmer RG (1989) Restriction fragment length polymorphism diversity in soybean. Theor Appl Genet 77:786–792
Kochert G, Halward T, Branch WD, Simpson CE (1990) RFLP variability in peanut (Arachis hypogaea L.) cultivars and wild species. Theor Appl Genet 81:565–570
McCouch SR, Kochert G, Yu ZH, Wang ZY, Khush GS, Coffman WR, Tanksley SD (1988) Molecular mapping of rice chromosomes. Theor Appl Genet 76:815–829
McGrath JM, Quiros CF, Harada JJ, Landry BS (1990) Identification of Brassica oleracea monosomic alien addition lines with molecular markers reveals extensive gene duplication. Mol Gen Genet 223:198–204
Michaud R, Lehman WF, Rumbaugh MD (1988) World distribution and historical development. In: Hanson AA, Barnes DK, Hill RR (eds) Alfalfa and alfalfa improvement. ASA, Madison/WI, pp 25–91
Moutray JB (1983) Registration of Apollo alfalfa. Crop Sci 23:178
Quiros CF (1983) Alfalfa, luzerne (Medicago sativa L.). In: Tanksley SD, Orton TJ (eds) Isozymes in plant genetics and breeding, Part B. Elsevier Scientific, Amsterdam, pp 253–294
Quiros CF, Bauchan GR (1988) The genus Medicago and the origin of the Medicago sativa complex. In: Hanson AA, Barnes DK, Hill RR (eds) Alfalfa and alfalfa improvement. ASA, Madison/WI, pp 93–124
Quiros CF, Morgan K (1981) Peroxidase and leucine-aminopeptidase in diploid Medicago species closely related to alfalfa: multiple gene loci, multiple allelism, and linkage. Theor Appl Genet 60:221–228
Rumbaugh MD, Caddel JL, Rowe DE (1988) Breeding and quantitative genetics. In: Hanson AA, Barnes DK, Hill RR (eds) Alfalfa and alfalfa improvement. ASA, Madison/WI, pp 777–808
Saghai-Maroof MA, Soliman KM, Jorgensen RA, Allard RW (1984) Ribosomal DNA spacer-length polymorphisms in barley: Mendelian inheritance, chromosomal location, and population dynamics. PNAS 81:8014–8018
Shattuck-Eidens DM, Bell RN, Neuhausen SL, Helentjaris T (1990) DNA sequence variation within maize and melon: observations from polymerase chain reaction amplification and direct sequencing. Genetics 126:207–217
Song K, Osborn TC, Williams PH (1990) Brassica taxonomy based on nuclear restriction fragment length polymorphisms (RFLPs). 3. Genome relationships in Brassica and related genera and the origin of B. oleracea and B. rapa (syn. campestris). Theor Appl Genet 79:497–506
Sorensen EL, Byers RA, Horber EK (1988) Breeding for insect resistance. In: Hanson AA, Barnes DK, Hill RR (eds) Alfalfa and alfalfa improvement, ASA, Madison/WI, pp 859–902
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Stanford EH (1951) Tetrasomic inheritance in alfalfa. Agron J 43:222–225
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Communicated by P. L. Pfahler
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Brummer, E.C., Kochert, G. & Bouton, J.H. RFLP variation in diploid and tetraploid alfalfa. Theoret. Appl. Genetics 83, 89–96 (1991). https://doi.org/10.1007/BF00229230
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DOI: https://doi.org/10.1007/BF00229230