Biopolym. Cell. 2005; 21(1):35-41.
Cell Biology
Intertribal somatic hybrids of di- and tetragenome origin: production
and usage
1Ovcharenko O. O., 1Komarnytskyy I. K., 1Cherep M. M., 1Rudas V. A., 1Kuchuk M. V.
- Institute of Cell Biology and Genetic Engineering, NAS of Ukraine
148, Akademika Zabolotnogo Str., Kyiv, Ukraine, 03680
Abstract
Intertribal hybrids in the combinations of Orychophragmus violaceus
(L.) O. E. Schulz. + Arabidopsis thaliana L. and O. violaceus (L.) O. E. Schulz. + hybrid (Brassica juncea (L.) Czern. & Coss + A.
thaliana L.) were obtained as a result of somatic hybridization.
Hybrid nature of the plants obtained was confirmed by the PCRRFLP and analysis of isozymes. A. thaliana L. plants were
transformed with a vector carrying heterologous transposable element SpmldSpm and genes nptll, bar, gus, which allowed to
examine the behavior of transgenic traits in digenomic and tetragenomic hybrids. The hybrid plants were similar to one of the
parental species (O. violaceus + A. thaliana) or had intermediate
morphology (O. violaceus + hybrid (B. juncea + A. thaliana)). All
the plants obtained were resistant to kanamycin and phosphinothricin due to the transgenes transferred from A. thaliana The
activity of β-glucuronidase was detected in hybrid O. violaceus +
+ A. thaliana while it was absent in the plants O. violaceus + hybrid
(B. juncea + A. thaliana). PCR analysis revealed maintenance of all
transgenes in the hybrids.
Keywords: intertribal somatic hybridization, Orychophragmus violaceus, Brassica juncea, Arabidopsis thaliana, heterologous system of transposable element Spm/dSpm
Full text: (PDF, in Ukrainian)
References
[1]
Gleba YuYu, Sitnik KM. Cell engineering of plants. Kiev, Naukova Dumka, 1984; 160 p.
[2]
Waara S, Glimelius K. The potential of somatic hybridization in crop breeding. Euphytica. 1995;85(1-3):217–33.
[3]
Vlahova M, Hinnisdaels S, Frulleux F, Claeys M, Atanassov A, Jacobs M. UV irradiation as a tool for obtaining asymmetric somatic hybrids between Nicotiana plumbaginifolia and Lycopersicon esculentum. Theor Appl Genet. 1997;94(2):184–91.
[4]
Bates GW, Hasenkampf CA, Contolini CL, Piastuch WC. Asymmetric hybridization in Nicotiana by fusion of irradiated protoplasts. Theor Appl Genet. 1987;74(6):718-26.
[5]
Menczel L, Morgan A, Brown S, Maliga P. Fusion-mediated combination of Ogura-type cytoplasmic male sterility with Brassica napus plastids using X-irradiated CMS protoplasts. Plant Cell Rep. 1987;6(2):98-101.
[6]
Dudits D, Fejer O, Hadlaczky G, Koncz C, Lazar GB, Horvath G. Intergeneric gene transfer mediated by plant protoplast fusion. Mol Gen Genet. 1980;179(2):283–8.
[7]
Sidorov VA, Menczel L, Nagy F, Maliga P. Chloroplast transfer in Nicotiana based on metabolic complementation between irradiated and iodoacetate treated protoplasts. Planta. 1981;152(4):341-5.
[8]
Ananiev EV, Riera-Lizarazu O, Rines HW, Phillips RL. Oat-maize chromosome addition lines: a new system for mapping the maize genome. Proc Natl Acad Sci U S A. 1997;94(8):3524-9.
[9]
Sundberg E, Glimelius K. Effects of parental ploidy level and genetic divergence on chromosome elimination and chloroplast segregation in somatic hybrids within Brassicaceae. Theor Appl Genet. 1991;83(1):81-8.
[10]
Babiychuk E, Kushnir S, Gleba YY. Spontaneous extensive chromosome elimination in somatic hybrids between somatically congruent species Nicotiana tabacum L. and Atropa belladonna L. Theor Appl Genet. 1992;84(1-2):87-91.
[11]
Tabaeizadeh Z, Perennes C, Bergounioux C. Increasing the variability of Lycopersicon peruvianum Mill. by protoplast fusion with Petunia hybrida L. Plant Cell Rep. 1985;4(1):7-11.
[12]
Turpin C. Attempt of male cytoplasmic sterility introduction by intergeneric fusion in cultivated tomato. Acta Hortic. 1986; 191: 377-9.
[13]
UN. Genome analysis in Brassica with special reference to the experimental formation of B. napus and peculiar mode to fertilization. Jap J Bot; 1935; 7(3):389-452.
[14]
Ovcharenko OO, Komarnyts'ky? IK, Cherep MM, Hleba IuIu, Kuchuk MV. [Obtaining of intertribal Brassica juncea+Arabidopsis thaliana somatic hybrids and study of transgenic trait behaviour]. Tsitol Genet. 2004;38(3):3-8.
[15]
Medgyesy P, Menczel L, Maliga P. The use of cytoplasmic streptomycin resistance: Chloroplast transfer from Nicotiana tabacum into Nicotiana sylvestris, and Isolation of their somatic hybrids. Mol Gen Genet. 1980;179(3):693–8.
[16]
Menczel L, Nagy F, Kiss ZR, Maliga P. Streptomycin resistant and sensitive somatic hybrids of Nicotiana tabacum + Nicotiana knightiana: correlation of resistance to N. tabacum plastids. Theor Appl Genet. 1981;59(3):191-5.
[17]
Sidorov VA, Zubko MK, Kuchko AA, Komarnitsky IK, Gleba YY. Somatic hybridization in potato: use of ?-irradiated protoplasts of Solanum pinnatisectum in genetic reconstruction. Theor Appl Genet. 1987;74(3):364-8.
[18]
Gamborg OL, Miller RA, Ojima K. Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res. 1968;50(1):151-8.
[19]
Jefferson RA. Assaying chimeric genes in plants: The GUS gene fusion system. Plant Mol Biol Rep. 1987;5(4):387–405.
[20]
Cheung WY, Hubert N, Landry BS. A simple and rapid DNA microextraction method for plant, animal, and insect suitable for RAPD and other PCR analyses. PCR Methods Appl. 1993;3(1):69-70.
[21]
Sakhno LA, Sytnik ES, Cherep NN, Komarnitski? IK, Kuchuk NV, Klimiuk VI. [Activity of the corn Spm transposon system in transgenic plants Orychophragmus violaceus (L.) O.E. Schulz obtained by both direct transfer of DNA to protoplasts and agrobacterial transformation of root explants]. Tsitol Genet. 2002;36(6):3-8.
[22]
White TJ, Bruns T, Lee S, Taylor J. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR protocols: a guide to methods and applications. Eds M. Innis et al. San Diego: Acad, press, 1990: 315-22.
[23]
Savolainen V, Corbaz R, Moncousin C, Spichiger R, Manen JF. Chloroplast DNA variation and parentage analysis in 55 apples. Theor Appl Genet. 1995;90(7-8):1138-41.
[24]
Tsumura Y, Kawahara T, Wickneswari R, Yoshimura K. Molecular phylogeny of Dipterocarpaceae in Southeast Asia using RFLP of PCR-amplified chloroplast genes. Theor Appl Genet. 1996;93(1-2):22-9.
[25]
Demesure B, Sodzi N, Petit RJ. A set of universal primers for amplification of polymorphic non-coding regions of mitochondrial and chloroplast DNA in plants. Mol Ecol. 1995;4(1):129-31.
[26]
Demeke T, Hucl P, Baga M, Caswell K, Leung N, Chibbar RN. Transgene inheritance and silencing in hexaploid spring wheat. Theor Appl Genet. 1999;99(6):947–53.
[27]
Bopisyuk NV, Klimyuk VI, Papokonnyy AS, Samoylov AM, Cherep NN, Shaxovskiy AM, Shlumukov LR. Biochemical analysis in cell engineering of plants. Kiev, 1988; 49 p. (Preprint. AN Ukr SSR, Institut botaniki; 88.1).
[28]
Gleba YuYu, Sytnik KM. ProtoPlast fusion and genetic engineering of higher Plants. Kyiv, Naukova Dumka, 1982; 104 p.
[29]
Tissier AF, Marillonnet S, Klimyuk V, Patel K, Torres MA, Murphy G, Jones JD. Multiple independent defective suppressor-mutator transposon insertions in Arabidopsis: a tool for functional genomics. Plant Cell. 1999;11(10):1841-52.
[30]
Vasilenko MIu, Komarnitski? IK, Sakhno LA, Gleba IuIu, Kuchuk NV. [Obtaining and analysis of intergeneric somatic hybrids between Brassica napus and "albino" line of Orychophragmus violaceus]. Tsitol Genet. 2003;37(1):3-10.
[31]
Sakhno LA, Vasilenko MYu, Ovcharenko OA, Kuchuk NV. Rapid plant regeneration fromleaf protoplastsand leaf petiole segments of Orychophragmus violaceus (L.) O. E. Schulz. Fiziologiia i biokhimiia kul'turnykh rasteniy. 2002; 34(1):75-9.
