Abstract
We examined genetic variation in allozyme loci, nuclear DNA restriction fragment length polymorphisms (RFLPs), and random amplified polymorphic DNAs (RAPDs) in 130 trembling aspen (Populus tremuloides) and 105 bigtooth aspen (P. grandidentata) trees. In trembling aspen 10 out of 13 allozyme loci assayed (77%) were polymorphic (P), with 2.8 alleles per locus (A) and an expected heterozygosity (He) of 0.25. In contrast, bigtooth aspen had a much lower allozyme genetic variability (P=29%; A=1.4; He=0.08). The two species could be distinguished by mutually exclusive alleles at Idh-1, and bigtooth aspen has what appears to be a duplicate 6PG locus not present in trembling aspen. We used 138 random aspen genomic probes to reveal RFLPs in HindIII digests of aspen DNA. The majority of the probes were from sequences of low copy number. RFLP results were consistent with those of the allozyme analyses, with trembling aspen displaying higher genetic variation than bigtooth aspen (P=71%, A=2.7, and He=0.25 for trembling aspen; P=65%, A=1.8, and He=0.13 for bigtooth aspen). The two species could be distinguished by RFLPs revealed by 21 probes (15% of total probes assayed). RAPD patterns in both species were studied using four arbitrary decamer primers that revealed a total of 61 different amplified DNA fragments in trembling aspen and 56 in bigtooth aspen. Assuming a Hardy-Weinberg equilibrium, estimates of P=100%, A=2, and He=0.30 in trembling aspen and P=88%, A=1.9, and He=0.31 in bigtooth aspen were obtained from the RAPD data. Five amplified DNA fragments were species diagnostic. All individuals within both species, except for 2 that likely belong to the same clone, could be distinguished by comparing their RAPD patterns. These results indicate that (1) RFLPs and allozymes reveal comparable patterns of genetic variation in the two species, (2) trembling aspen is more genetically variable than bigtooth aspen at both the allozyme and DNA levels, (3) one can generate more polymorphic and species-specific loci with DNA markers than with allozymes in aspen, and (4) RAPDs provide a very powerful tool for “fingerprinting” aspen individuals.
Similar content being viewed by others
References
Adams WT (1983) Application of isozymes in tree breeding. In: Tanksley SD, Orton TJ (eds) Isozymes in plant genetics and breeding, part A. Elsevier Science Publ. Amsterdam, pp 381–400
Andrejak GE, Barnes BV (1969) A seedling population of aspens in southeastern Michigan. Mich Bot 8:189–202
Ayala FJ (1983) Enzymes as taxonomic characters. In: Oxford GS, Rollonson D (eds) Protein polymorphism: adaptive and taxonomic significance. (The Systematics Association Special Volume No. 24.) Academic Press, New York, pp 3–26
Barnes BV (1961) Hybrid aspens of the Lower Peninsula of Michigan. Rhodora 63:311–324
Barnes BV (1969) Natural variation and delineation of clones of Populus tremuloides and P. grandidentata in northern lower Michigan. Silvae Genet 18:130–142
Barnes BV (1975) Phenotypic variation of trembling aspen of western North America. For Sci 21:319–328
Beckmann JS, Soller M (1986) Restriction fragment length polymorphisms and genetic improvement of agricultural species. Euphytica 35:111–124
Burr B, Evola SV, Burr FA, Beckmann JS (1983) The application of restriction fragment length polymorphisms to plant breeding. In: Setlow JK, Hollander A (eds) Genetic engineering principles and methods, vol 5. Plenum Press, New York London, pp 45–59
Chase CD, Ortega VM, Vallejos CE (1991) DNA restriction fragment length polymorphisms correlate with isozyme diversity in Phaseolus vulgaris L. Theor Appl Genet 81:806–811
Cheliak WM, Dancik BP (1982) Genetic diversity of natural populations of a clone-forming tree Populus tremuloides. Can J Genet Cytol 24:611–616
Cheliak WM, Pitel JA (1984) Electrophoretic identification of clones in trembling aspen. Can J For Res 14:740–743
Clegg MT (1989) Molecular diversity in plant populations. In: Brown AHD, Clegg MT, Kahler AL, Weir BS (eds) Plant population genetics, breeding, and genetic resources. Sinauer Assoc, Sunderland, Mass., pp 98–115
Devey ME, Jermstad KD, Tauer CG, Neale DB (1991) Inheritance of RFLP loci in a loblolly pine three-generation pedigree. Theor Appl Genet 83:238–242
Edwards M (1992) Use of molecular markers in the evaluation and introgression of genetic diversity for quantitative traits. Field Crops Res 29:241–260
Einspahr DW, Joranson PN (1960) Late flowering in aspen and its relation to naturally occurring hybrids. For Sci 6:221–223
Farmer MM, Barnes BV (1978) Morphological variation of families of trembling aspen in southeastern Michigan. Mich Bot 17:141–153
Halward TM, Stalker HT, La Rue EA, Kochert G (1991) Genetic variation detectable with molecular markers among unadapted germ-plasm resources of cultivated peanut and related wild species. Genome 34:1013–1020
Hamrick JL, Godt MJW (1989) Allozyme diversity in plant species. In: Brown AHD, Clegg MT, Kahler AL, Weir BS (eds) Plant population genetics, breeding, and genetic resources. Sinauer Assoc., Sunderland, Mass., pp 43–63
Helentjaris T, King G, Slocum M, Siedenstrang C, Wegmann S (1985) Restriction fragment polymorphisms as probes for plant diversity and their development as tools for applied plant breeding. Plant Mol Biol 5:109–118
Henry RM, Barnes BV (1977) Comparative reproductive ability of bigtooth and trembling aspen and their hybrids. Can J Bot 55:3093–3098
Hocking RR (1985) The analysis of linear models. Brooks/Cole Publ, Monterey, Calif.
Hospital F, Chevalet C, Mulsant P (1992) Using markers in gene introgression breeding programs. Genetics 132:1199–1210
Hyun JO, Rajora OP, Zsuffa L (1987) Genetic variation in trembling aspen in Ontario based on isozyme studies. Can J For Res 17:1134–1138
Jelinski DE, Cheliak WM (1992) Genetic diversity and spatial subdivision of Populus tremuloides (Salicaceae) in a heterogeneous landscape. Am J Bot 79:728–736
Kemperman JA, Barnes BV (1976) Clone size in American aspens. Can J Bot 54:2603–2607
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
Laidly PR (1990) Populus grandidentata Michx. Bigtooth aspen In: Burns RM, Honkala BH (ededs) Silvics of North America, vol 2; hardwoods. USDA For Sev Agric Handb 654, pp 544–550
Liu Z, Furnier GR (1993) Inheritance and linkage of allozymes and RFLPs in trembling aspen. J Hered (in press)
Lund ST, Furnier GR, Mohn CA (1992) Isozyme variation in quaking aspen in Minnesota. Can J For Res 22:521–524
McComb AL, Hanson N (1954) A naturally occurring aspen popular hybrid. J For 52:528
McDonough WT (1985) Sexual reproduction, seeds, and seedlings. In: DeByle NV, Winokur RP (eds) Aspen: ecology and management in the western United States. USDA For Serv Gen Tech Rep RM-119, pp 25–33
McGrath JM, Quiros CF (1992) Genetic diversity at isozyme and RFLP loci in Brassica campestris as related to crop type and geographical origin. Theor Appl Genet 83:783–790
Messmer MM, Melchieger AE, Lee M, Woodman WL, Lee EA, Lamkey KR (1991) Genetic diversity among progenitors and elite lines from the Iowa Stiff Stalk Synthetic (BSSS) maize populations: comparison of allozyme and RFLP data. Theor Appl Genet 83:97–107
Mitton JB, Grant MC (1980) Observations on the ecology and evolution of quaking aspen, Populus tremuloides, in the Colorado Front Range. Am J Bot 67:202–209
Muller HJ (1964) The relation of recombination to mutation advance. Mutat Res 1:2–9
Nei M (1978) Estimation of average heterozygosity and genetic distance from a small number of individuals. Genetics 89:583–590
Neuhausen SL (1992) Evaluation of restriction fragment length polymorphism in Cucumis melo. Theor Appl Genet 83:379–384
Pauley SS (1956) Natural hybridization of the aspens. Univ Minn For Notes No. 47
Perala DA (1990) Populus tremuloides Michx. Quaking aspen. In: Burns RM, Honkala BH (eds) Silvics of North America, vol 2: hardwoods. USDA For Serv Agric Hand. 654, pp 555–569
Quiros CF, Hu J, This P, Chevre AM, Delseny M (1991) Development and chromosomal localization of genome-specific markers by polymerase chain reaction in Brassica. Theor Appl Genet 82:627–632
Rajora OP (1990) Marker allozyme genes and alleles for differentiation of Populus deltoides, P. nigra, P. maximowiczii, and their interspecific hybrids. Can J Bot 68:990–998
Rogstad SH, Nybom H, Schaal B (1991) The tetrapod “DNA fingerprinting” M 13 repeat probe reveals genetic diversity and clonal growth in quaking aspen (Populus tremuloides, Salicaceae). Plant Syst Evol 175:115–123
Snedecor GW, Cochran WG (1967) Statistical methods, 6th edn. Iowa State University Press, Ames, Iowa
Spencer JS Jr, Leatherberry EC, Kingsley NP (1990) The Lake States' aspen resource revisited: mid-1960s-1987. In: Adams RD (ed) Proc Aspen Symp '89. USDA For Serv Gen Tech Rep NC-140, pp 243–252
Wilde J, Waugh R, Powell W (1992) Genetic fingerprinting of Theobroma clones using randomly amplified polymorphic DNA markers. Theor Appl Genet 83:871–877
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
Zamir D, Tanksley SD (1988) Tomato genome is comprised largely of fast-evolving, low copy-number sequences. Mol Gen Genet 213:254–261
Zhang Q, Saghai-Maroof MA, Liu TY, Shen BZ (1992) Genetic diversity and differentiation of indica and japonica rice detected by RFLP analysis. Theor Appl Genet 83:495–499
Author information
Authors and Affiliations
Additional information
Communicated by P. M. A. Tigerstedt
Rights and permissions
About this article
Cite this article
Liu, Z., Furnier, G.R. Comparison of allozyme, RFLP, and RAPD markers for revealing genetic variation within and between trembling aspen and bigtooth aspen. Theoret. Appl. Genetics 87, 97–105 (1993). https://doi.org/10.1007/BF00223752
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00223752