Hereditary Behavior of bar Gene Cassette is Complex in Rice Mediated by Particle Bombardment

a.
College of Food, Biological and Environmental Engineering, Zhejiang Gongshang University, Hangzhou 310035, China
b.
State Key Laboratory of Rice Biology, China National Rice Research Institute, Hangzhou 310006, China
c.
Life Science Department of Hangzhou Teacher's College, Hangzhou 310036, China

More Information

Corresponding author: E-mail address: qianqian188@163.com (Qian Qian)

摘要

- /
- /
Abstract: Particle bombardment transformation using minimal gene cassette (containing the promoter, open reading frame and terminator) is the novel trend in plant genetic transformation, and its use helps to alleviate the undesirable effects of plasmid vector backbone sequences on transgenic plants. In the present article, studies related to the hereditary behavior of bar gene cassette in T₁ to T₃ generations of the transgenic rice (Oryza sativa L.) lines transformed by particle bombardment have been discussed. The selectable marker bar gene cassette that integrated with the rice genome had multiple copies and showed complex segregation behaviors including the presence of ‘false homozygotes’, with abnormal segregation ratios ranging from 35:1 to 144:1 (Basta-resistant: sensitive plants) in their progenies. In five out of ten original transgenic lines, bar gene can be stably transmitted as a dominant gene to self-pollinated T₂ progeny. The homozygotes were obtained in three transgenic lines in T₁ generation regardless of the multiple-copy integration patterns ofbar gene. Southern blotting analysis showed that multiple copies of bar gene cassette were linked, which formed transgene arrays in the host rice genome. The authors also observed stable transmission of integration patterns of bar gene cassette, as obtained from Southern blotting analysis, in the regularly segregated transgenic rice lines and loss of gene in an irregularly segregated transgenic line. The segregation behavior varied among the transgenic progenies that exhibited similar Southern hybridization patterns of bar gene. On the basis of these results, the multiple-copy integration, gene lost, and gene expression interaction were the major reasons for the complex segregation behaviors of bar gene cassette in transgenic rice plants.
- particle bombardment /
- hereditary behavior /
- rice

HTML全文

参考文献(24)

[1]	D, Gahakwa, SB, et al. Transgenic rice as a system to study the stability of transgene expression: multiple heterologous transgenes show similar behavior in diverse genetic backgrounds Theor Appl Genet, 101 (2000),pp. 388-399
[2]	J, Jakowitsch, I, et al. Molecular and cytogenetic characterization of a transgene locus that induces silencing and methylation of homologous promoters in trans Plant J, 17 (1999),pp. 131-140
[3]	AE, Muller, Y, et al. J Mol Biol, 291 (1999),pp. 29-46
[4]	XD, Fu, LT, et al. Linear transgene constructs lacking vector backbone sequences generate low-copy-number transgenic plants with simple integration patterns Transgenic Res, 9 (2000),pp. 11-19
[5]	JC, Breitler, A, et al. Efficient microprojectile bombardment-mediated transformation of rice using gene cassettes Theor Appl Genet, 104 (2002),pp. 709-719
[6]	P, Holger Towards the ideal GMP: Homologous recombination and marker gene excision J Plant Physiol, 160 (2003),pp. 743-754
[7]	F, Altpeter, N, et al. Particle bombardment and the genetic enhancement of crops: myths and realities Mol Breed, 15 (2005),pp. 305-332
[8]	NT, Loc, P, et al. Linear transgene constructs lacking vector backbone sequences generate transgenic rice plants which accumulate higher levels of proteins conferring insect resistance Mol Breed, 9 (2002),pp. 231-244
[9]	Y, Zhao, YC, et al. Acta Genet Sin, 30 (2003),pp. 135-141
[10]	A, Romano, K, et al. Transgene organization in potato after particle bombardment-mediated (co)transformation using plasmids and gene cassettes Transgenic Res, 12 (2003),pp. 461-473
[11]	Q, Yao, L, et al. Inheritance of the foreign gene cassettes lacking vector backbone sequences Hereditas, 28 (2006),pp. 695-698
[12]	JR, Vidal, JR, et al. Biolistic transformation of grapevine using minimal gene cassette technology Plant Cell Rep, 25 (2006),pp. 807-814
[13]	DN, Huang, B, et al. Sci China (Ser. C), 39 (1996),pp. 652-661
[14]	YJ, Lu, KL, et al. A simple method for isolation of rice DNA Chinese Journal of Rice Science, 6 (1992),pp. 47-48
[15]	J, Sambrook, EF, et al.
[16]	JY, Peng, FJ, et al. Plant Mol Biol, 27 (1995),pp. 91-104
[17]	J, Cooley, T, et al. Molecular and genetic characterization of elite transgenic rice plants produced by electric-discharged particle acceleratin Theor Appl Genet, 90 (1995),pp. 97-104
[18]	L, Wu, S, et al. Expression and inheritance of nine transgenes in rice Transgenic Res, 11 (2002),pp. 533-541
[19]	ZS, Tang, XK, et al. Acta Genet Sin, 27 (2000),pp. 26-33
[20]	F, Goto, S, et al. Inheritance of a co-transerred foreign gene in the progenies of transgenic plants Transgenic Res, 2 (1993),pp. 302-305
[21]	P, Vain, A, et al. Trangene behaviour across two generations in a large random population of transgenic rice plants produced by particle bombardment Theor Appl Genet, 105 (2002),pp. 878-889
[22]	XB, Xie, HR, et al. Studies on the distorted segregation of foreign genes in transgenic rice progenies Acta Genet Sin, 29 (2002),pp. 1005-1011
[23]	A, Kohli, M, et al. Transgene organization in rice engineered through direct DNA transfer supports a two-phase mechanism mediated by the establishment of integration hot sports Proc Natl Acad Sci USA, 95 (1998),pp. 7203-7208
[24]	A, Kohli, RM, et al. Transgene integration, organization and interaction in plants Plant Mol Biol, 52 (2003),pp. 247-258