Abstract
Five populations of lemon plants [Citrus limon (L.) Burm] obtained from undeveloped ovules through different tissue culture procedures were examined for the presence of somaclonal and irradiation-induced genetic variation. Tested groups were: (1) nucellar seedlings; (2) organogenic, regenerated via adventitious buds from nucellar seedling internodes; (3) embryogenic population, regenerated from non-irradiated nucellar callus via somatic embryogenesis; (4) embryogenic population, regenerated from irradiated nucellar callus via somatic embryogenesis; and (5) protoplast-derived, regenerated via somatic embryogenesis. Genomic DNA samples from 360 plants (72 from each group) were screened for polymorphism among RAPD fingerprints amplified by 10 decamer primers. Among all tested plants, genetic variation was detected only within the group of plants recovered from irradiated embryogenic calli. Out of 72 plants from that group, three had RAPD fingerprints different from the rest of the population, and fourth plant was found to be cytochimeric, consisting of diploid and tetraploid cells as revealed by flow cytometry. In all other populations of regenerated plants, we did not come across any plants with changed ploidy level.
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Abbreviations
- BA:
-
6-Benzyladenine
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
- MES:
-
4-Morpholinoethanesulfonic acid
References
Al-Zahim MA, Ford-Lloyd BV, Newbury HJ (1999) Detection of somaclonal variation in garlic (Allium sativum L.) using RAPD and cytological analysis. Plant Cell Rep 18:473–477
Arihara A, Kita T, Igarashi S, Goto M, Irikura W (1995) White Baron—a non browning somaclonal variant of Danshakuimo (Irish cobbler). Amer Potato J 72:701–705
Ćalović M, Viloria Z, Nielsen B, Gmitter Jr FG, Castle WS, Grosser JW (2004) Somatic embryogenesis from lemon styles and analysis of genetic stability in regenerated plants using RAPD and flow cytometry. Proc Int Soc Citricult IX Congr 2000, Orlando, FL, pp 131–134
Deng XX, Grosser JW, Gmitter Jr FG (1992) Intergeneric, somatic hybrid plants from protoplast fusion of Fortunella crassifolia cultivar ‘Meiwa’ with Citrus sinensis cultivar ‘Valencia’. Sci Hort 49:55–62
Engelborghs I, Swennen R, van Campenhout S (1998) The potential of AFLP to detect genetic differences in somaclonal variants in Musa sp Infomusa 7:3–6
Fourré JL, Berger P, Niquet L, Andre P (1997) Somatic embryogenesis and somaclonal variation in Norway spruce: morphogenetic, cytogenetic and molecular approaches. Theor Appl Genet 94:159–169
Gao MW, Cheng XY, Liang ZQ (1991) Generating pattern and characteristics of wheat somaclonal variation in the first generation. In: Chen Y et al (ed) Plant somaclonal variation and breeding. Jiangsu Sci Tech Pub House, China, pp 50–59
Gmitter Jr FG, Moore GA (1986) Plant regeneration from undeveloped ovules and embryogenic calli of Citrus: embryo production, germination, and plant survival. Plant Cell Tiss Org Cult 6:139–147
Grosser JW, Gmitter Jr FG (1990) Protoplast fusion and citrus improvement. Plant Breeding Rev 8:339–374
Grosser JW, Gmitter Jr FG, Chandler JL (1996) Somaclonal variation in sweet orange—a tool for cultivar improvement. Proc Int Soc Citricult VIII Congr, Sun City, South Africa, pp 203–206
Hashmi G, Huettel R, Meyer R, Krusberg L, Hammerschlag F (1997) RAPD analysis of somaclonal variants derived from embryo callus cultures of peach. Plant Cell Rep 16:624–627
Heinze B, Schmidt J (1995) Monitoring genetic fidelity vs somaclonal variation in Norway spruce (Picea abies) somatic embryogenesis by RAPD analysis. Euphytica 85:341–345
Jain SM (2001) Tissue culture-derived variation in crop improvement. Euphytica 118:153–166
Loureiro J, Pinto G, Lopes T, Doležel J, Santos C (2005) Assessment of ploidy stability of the somatic embryogenesis process in Quercus suber L. using flow cytometry. Planta 221(6):815–822
Mo LH, von Arnold S, Lagercrantz U (1989) Morphogenic and genetic stability in longterm embryogenic cultures and somatic embryos of Norway spruce (Picea abies L. Karst). Plant Cell Rep 8:375–378
Moretzsohn MC, Hopkins MS, Mitchell SE, Kresovich S, Valls JF, Ferreira ME (2004) Genetic diversity of peanut (Arachis hypogaea L.) and its wild relatives based on the analysis of hypervariable regions of the genome. BMC Plant Biol. 4:11, (electronic resource)
Murashige T, Tucker DPH (1969) Growth factor requirements of citrus tissue culture. Proc First Int Citrus Symp 3:1155–1161
Orbović V, Grosser JW (2006) Citrus: sweet orange (Citrus sinensis L. Osbeck ‘Valencia’) and Carrizo citrange [Citrus sinensis (L.) Osbeck × Poncirus trifoliata (L.) Raf.]. In: Wang K (ed) Agrobacterium protocols-methods in molecular biology. Humana Press Inc, USA, pp 177–189
Pfosser M, Amon A, Lelley T, Heberle-Bors E (1995) Evaluation of sensitivity of flow cytometry in detecting aneuploidy in wheat using disomic and ditelosomic wheat-rye addition lines. Cytometry 21:387–393
Rival A, Beule T, Barre P, Hamon S, Duval Y, Noirot M (1997) Comparative flow cytometric estimation of nuclear DNA content in oil palm (Elaeis guineensis Jacq) tissue cultures and seed-derived plants. Plant Cell Rep 16:884–887
Veilleux RE, Johnson AAT (1998) Somaclonal variation: molecular analysis, transformation, interaction, and utilization. Plant Breed Rev 16:229–268
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
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Orbović, V., Ćalović, M., Viloria, Z. et al. Analysis of genetic variability in various tissue culture-derived lemon plant populations using RAPD and flow cytometry. Euphytica 161, 329–335 (2008). https://doi.org/10.1007/s10681-007-9559-3
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DOI: https://doi.org/10.1007/s10681-007-9559-3