Korean Journal of Breeding Science (한국육종학회지)

The Korean Society of Breeding Science (한국육종학회)
권호 표지

Karyotype Analyses of a Rice Cultivar 'Nakdong' and its Four Genetically Modified Events by Conventional Staining and Fluorescence in situ Hybridization

  • Jeon, Eun Jin (Plant Biotechnology Institute, Department of Life Science, Sahmyook University) ;
  • Ryu, Kwang Bok (Plant Biotechnology Institute, Department of Life Science, Sahmyook University) ;
  • Kim, Hyun Hee (Plant Biotechnology Institute, Department of Life Science, Sahmyook University)
  • Received : 2011.05.13
  • Published : 2011.09.30
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Abstract

Conventional staining and fluorescence in situ hybridization (FISH) karyotypes of the non-genetically modified (GM) parental rice line, 'Nakdong' (Oryza sativa L. japonica), and its four GM rice lines, LS28 (event LS30-32-20-1), Cry1Ac1 (event C7-1-9-1), and LS28 ${\times}$ Cry1Ac1 (events L/C1-1-3-1 and L/C1-3-1-1) were analyzed using 5S and 45S rDNAs as probes. Both parental and transgenic lines were diploids (2n=24) with one satellite chromosome pair. The lengths of the prometaphase chromosomes ranged from 1.50 to $6.30{\mu}m$. Four submetacentric and eight metacentric pairs comprised the karyotype of 'Nakdong' and its four GM lines. One pair of 5S rDNA signals was detected near the centromeric region of chromosome g in both the parental and transgenic lines. The 45S rDNA signals were detected on the secondary constrictions of the satellite chromosome pair in both the parental and transgenic lines. There was no significant difference in chromosome size, length, and composition between 'Nakdong' and its four GM lines. This research was conducted as a preliminary study for chromosomal detection of transgenes in GM rice lines and would be useful for their breeding programs.

Keywords

Acknowledgement

Supported by : Sahmyook University

References

  1. Arumuganathan K, Earle ED. 1991. Nuclear DNA content of some important plant species. Plant Mol. Biol. Rep. 9:208-218. https://doi.org/10.1007/BF02672069
  2. Ansari A, Gartenberg MR. 1999. Persistence of an alternate chromatin structure at silenced loci in vitro. Proc. Natl. Acad. Sci. 96:343-348. https://doi.org/10.1073/pnas.96.2.343
  3. Baum B, Bailey G. 2001. The 5S rDNA gene sequence variation in wheats and some polyploidy wheat progenitors (Poaceae: Triticeae). Gen. Res. Crop Evol. 48:35-51. https://doi.org/10.1023/A:1011263107219
  4. Buell CR, Yu Y, Wing RA, Nelson W, de la Bastide M, Minx P, Jin WW, Jackson S. et al. The Rice Chromosome 3 Sequencing Consortium. 2005. Sequence, annotation, and analysis of synteny between rice chromosome 3 and diverged grass species. Genome Res. 15:1284-1291. https://doi.org/10.1101/gr.3869505
  5. Cheng Z, Buell CR, Wing RA, Gu M. Jiang J. 2001. Toward a cytological characterization of the rice genome. Genome Res. 11:2133-2141. https://doi.org/10.1101/gr.194601
  6. Choi HW, Lemaux PG, Cho M-J. 2000. Increased chromosomal variation in transgenic versus nontransgenic barley (Hordeum vulgare L.) plants. Crop Sci. 40:524-533. https://doi.org/10.2135/cropsci2000.402524x
  7. Choi HW, Lemaux PG, Cho MJ. 2001. High frequency of cytogenetic aberration in transgenic oat (Avena sativa L.) plants. Plant Sci. 156:85-94.
  8. Chu YE. 1967. Pachytene analysis and observation of chromosome association in haploid rice. Cytologia 32:82-95.
  9. Chung M-C, Lee Y-I, Cheng Y-Y, Chou Y-J, Lu C-F. 2008. Chromosomal polymorphism of ribosomal genes in the geneus Oryza. Theor. Appl. Genet. 116:745-753. https://doi.org/10.1007/s00122-007-0705-z
  10. de Melo NF, Guerra M. 2003. Variability of the 5S and 45S rDNA sites in Passiflora L. species with distinct base chromosome numbers. Ann. Bot. 92:309-316. https://doi.org/10.1093/aob/mcg138
  11. Devi J, Ko JM, Seo BB. 2005. FISH and GISH: Modern cytogenetic techniques. Indian J. Biotechnol. 4:307-315.
  12. Fukui K, Iijima K. 1991. Somatic chromosome map of rice by imaging methods. Theor. Appl. Genet. 81:589-596.
  13. Fukui K, Ohmido N, Khush GS. 1994. Variability in rDNA loci in the genus Oryza detected through fluorescence in situ hybridization. Theor. Appl. Genet. 87:893-899.
  14. Gerlach WL, Bedbrook JR. 1979. Cloning and characterization of ribosomal rDNA genes from wheat and barley. Nucleic Acids Res. 7:1869-1885. https://doi.org/10.1093/nar/7.7.1869
  15. Goff SA. 1999. Rice as a model for cereal genomics. Curr. Opin. Plant Biol. 2:86-89. https://doi.org/10.1016/S1369-5266(99)80018-1
  16. Han YH, Zhang ZH, Liu JH, Lu JY, Huang SW, Jin WW. 2008. Distribution of the tandem repeat sequences and karyotyping in cucumber (Cucumis sativus L.) by fluorescence in situ hybridization. Cytogenet. Genome Res. 122:80-88. https://doi.org/10.1159/000151320
  17. Hanson RE, Islam-Faridi MN, Percival EA, Crane CF, Ji Y, McKnight TD, Stelly DM, Price HJ. 1996. Distribution of 5S and 18S-28S rDNA loci in a tetraploid cotton (Gossypium hirsutum L.) and its putative diploid ancestors. Chromosoma 105:55-61. https://doi.org/10.1007/BF02510039
  18. Hu C-H, Kao K-N, Chang C-C. 1964. Further studies on the chromosome morphology of Oryza sativa L. Rice genetics and cytogenetics. p. 51-61.
  19. Hwang YJ, Lee SN, Song KA, Ryu KB, Ryu KH, Kim HH. 2010. Karyotype analyses of genetically modified (GM) and non-GM hot peppers by conventional staining and FISH method. Hort. Environ. Biotechnol. 51:525-530.
  20. Iglesias VA, Moscone EA, Papp I, Neuhuber F, Michalowski S, Phelan T, Spiker S, Matzke M, Matzke AJ. 1997. Molecular and cytogenetic analyses of stably and unstably expressed transgene loci in tobacco. Plant Cell 9:1251-1264.
  21. Ishiki K. 1985. Cytogenetical studies on African rice, Oryza glaberrima. Steud. 1. Colchicine-induced autotetraploid. Jpn. J. Genet. 60:225-234. https://doi.org/10.1266/jjg.60.225
  22. Islam-Faridi MN, Ishii T, Kumar V, Sitch LA, Brar DS. 1990. Chromosomal location of ribosomal RNA genes in rice by in situ hybridization. Rice Genet. Newslett. 7:143-144.
  23. Jiang J, Gill BS, Wang G-L, Ronald PC, Ward DC. 1995. Metaphase and interphase fluorescence in situ hybridization mapping of the rice genome with bacterial artificial chromosomes. Proc. Natl. Acad. Sci. 92:4487-4491. https://doi.org/10.1073/pnas.92.10.4487
  24. Jin WW, Li ZY, Fang Q, Altosaar I, Liu LH, Song YC. 2002. Fluorescence in situ hybridization analysis of alien genes in Agrobacterium-mediated Cry1A(b)-transformed rice. Ann. Bot. 90:31-36. https://doi.org/10.1093/aob/mcf160
  25. Kato A, Vega JM, Han F, Lamb JC, Birchler JA. 2005. Advances in plant chromosome identification and cytogenetic techniques. Curr. Opin. Plant Biol. 8:148-154. https://doi.org/10.1016/j.pbi.2005.01.014
  26. Kim J-H, Song H-S, Jee S-M, Ryu T-H, Kim D-H, Kim H-Y. 2005. Qualitative PCR detection of GM rices (Milyang 204 and Iksan 483) developed in Korea. J. Korean Soc. Appl. Biol. Chem. 48:335-338.
  27. Kim HJ, Song NH, Kang SG. 1988. Karyotype analysis of Oryza sativa cv. cheongcheong and it's autotetraploid induced by colchicine. Korean J. Genetics 10:175-183.
  28. Koo DH, Choi HW, Cho J, Hur Y, Bang JW. 2005. A high-resolution karyotype of cucumber (Cucumis sativus L. 'Winter Long') revealed by C-banding, pachytene analysis, and RAPD-aided fluorescence in situ hybridization. Genome 48:534-540. https://doi.org/10.1139/g04-128
  29. Kurata N, Omura T. 1978. Karyotype analysis in rice I. a new method for identifying all chromosome pairs. Japan J. Genetics 4:251-255.
  30. Kuwada Y. 1910. A cytological study of Oryza sativa L. Bot. Mag. (Tokyo) 24:267-281.
  31. Lee JS, Seo BB, Kim M. 2010. Karyotype analysis and rDNA physical mapping in rye (Secale cereal L.). Korean J. Breed. Sci. 42:163-168.
  32. Levan A, Fredga K, Sandberg AA. 1964. Nomenclature for centromeric position on chromosomes. Hereditas, 52:201-220.
  33. Li Z-Y, Fu M-L, Hu F-F, Huang S-F, Song Y-C. 2006. Visualization of the ribosomal DNA (45S rDNA) of Indica rice with FISH on some phases of cell cycle and extended DNA fibers. Biocell 30:27-32.
  34. Lim K-B, de Jong H, Yang T-J, Park J-Y, Kwon S-J, Kim J-S, Lim M-H, Kim JA, Jin M, Jin Y-M, Kim SH, Lim YP, Bang J-W, Kim H-I, Park B-S. 2005. Characterization of rDNAs and tandem repeats in the heterochromatin of Brassica rapa. Mol. Cells 19:436-444.
  35. Matzke AJ, Matzke MA. 1998. Position effects and epigenetic silencing of plant transgenes. Curr. Opin. Plant Biol. 1:142-148. https://doi.org/10.1016/S1369-5266(98)80016-2
  36. Matzke MA, Moscone EA, Park Y-D, Papp I, Oberkofler H, Neuhuber F, Matzke AJM. 1994. Inheritance and expression of a transgene insert in an aneuploid tobacco line. Mol. Gen. Genet. 245:471-485. https://doi.org/10.1007/BF00302260
  37. Ohmido N, Akiyama Y, Fukui K. 1998. Physical mapping of unique nucleotide sequences on identified rice chromosomes. Plant Mol. Biol. 38:1043-1052. https://doi.org/10.1023/A:1006062905742
  38. Park HM, Jeon EJ, Waminal NE, Shin KS, Kweon SJ, Park B-S, Suh SC, Kim HH. 2010. Detection of transgenes in three genetically modified rice lines by fluorescence in situ hybridization. Genes and Genomics. 32:527-531. https://doi.org/10.1007/s13258-010-0064-z
  39. Rau NS. 1929. Further contributions to the cytology of some crop plants of south India. J. Indian Bot. Soc. 81:201-206.
  40. Shastry SVS, Ranga Rao DR, Misra RN. 1960. Pachytene analysis in Oryza. I. Chromosome morphology in Oryza sativa. Indian J. Genet. and Plant Breed. 20:15-21.
  41. Shin K-S, Lee S-M, Lim S-H, Woo H-J, Cho H-S, Lee K-R, Lee M-C, Kweon S-J, Suh S-C. 2009. Molecular biological characteristics and analysis using the specific markers of leaf folder-resistant GM rice. J. Plant Biotechnol. 36:115-123. https://doi.org/10.5010/JPB.2009.36.2.115
  42. Shin K-S, Park J-H, Lee J-H, Lee S-M, Woo H-J, Lim S-H, Kim H-Y, Suh S-C and Kweon S-J. 2009a. Qualitative PCR detection of stack gene GM rice ($LS28{\times}Cry1Ac$) developed in Korea. J. Appl. Biol. Chem. 52:1-7. https://doi.org/10.3839/jabc.2009.001
  43. Shin YB, Katayama T. 1979. Studies on officinalis chromosome addition lines to cultivated rice, Oryza sativa L. Bull. Inst. Trop. Agr. Kyushu Univ. 3:1-60.
  44. Shishido R, Sano Y, Fukui K. 2000. Ribosomal DNAs: an exception to the conservation of gene order in rice genomes. Mol. Gen. Genet. 263:586-591. https://doi.org/10.1007/s004380051205
  45. Singh RJ, Klein TM, Mauvais CJ, Knowlton S, Hymowitz T, Kostow CM. 1998. Cytological characterization of transgenic soybean. Theor. Appl. Genet. 96:319-324. https://doi.org/10.1007/s001220050743
  46. Tao Q, Chang YL, Wang J, Chen H, Islam-Faridi MN, Scheuring C, Wang B, Stelly DM, Zhang HB. 2001. Bacterial Artificial Chromosome-Based Physical Map of the Rice Genome Constructed by Restriction Fingerprint Analysis. Genetics. 158:1711-1724.
  47. Vaughan DA, Morishima H, Kadowaki K. 2003. Diversity in the Oryza genus. Curr. Opin. Plant Biol. 6:139-146. https://doi.org/10.1016/S1369-5266(03)00009-8
  48. Wako T, Furushima-Shimogawara R, Belyaev ND, Turner BM, Fukui K. 2002. Cell cycle dependent and lysine residue-specific dynamic changes of histone H4 acetylation in barley. Plant Mol. Biol. 49:645-653.
  49. Wako T, Murakami Y, Fukui K. 2005. Comprehensive analysis of dynamics of histone H4 acetylation in mitotic barley cells. Genes Genet. Syst. 80:269-276. https://doi.org/10.1266/ggs.80.269
  50. Wang X, He C, Moore SC, Ausio J. 2001. Effects of histone acetylation on the solubility and folding of the chromatin fiber. J. Biol. Chem. 276:12764-12768. https://doi.org/10.1074/jbc.M100501200
  51. Xu YH, Yang F, Cheng YL, Ma L, Wang JB, Li LJ. 2007. Comparative analysis of rDNA distribution in metaphase chromosomes of Cucurbitaceae species. Hereditas (Beijing) 29:614-620. https://doi.org/10.1360/yc-007-0614
  52. Zhu X-Y, Cai D-T, Ding Y. 2008. Molecular and cytological characterization of 5S rDNA in Oryza species: genomic organization and phylogenetic implications. Genome 51:332-340. https://doi.org/10.1139/G08-016