Chromosomal translocations are a common phenomenon in Arabidopsis thaliana T-DNA insertion lines

被引:75
作者
Clark, Katie A.
Krysan, Patrick J. [1 ]
机构
[1] Univ Wisconsin Madison, Genome Ctr Wisconsin, Madison, WI 53706 USA
基金
美国国家科学基金会;
关键词
T-DNA; translocation; chromosome; Arabidopsis; pollen lethality; ILLEGITIMATE RECOMBINATION; INTEGRATION; TRANSFORMATION; MUTANTS; GENES;
D O I
10.1111/j.1365-313X.2010.04386.x
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
P>Ordered collections of Arabidopsis thaliana lines containing mapped T-DNA insertions have become an important resource for plant scientists performing genetic studies. Previous reports have indicated that T-DNA insertion lines can have chromosomal translocations associated with the T-DNA insertion site, but the prevalence of these rearrangements has not been well documented. To determine the frequency with which translocations are present in a widely-used collection of T-DNA insertion lines, we analyzed 64 independent lines from the Salk T-DNA mutant collection. Chromosomal translocations were detected in 12 of the 64 lines surveyed (19%). Two assays were used to screen the T-DNA lines for translocations: pollen viability and genome-wide genetic mapping. Although the measurement of pollen viability is an indirect screen for the presence of a translocation, all 11 of the T-DNA lines showing an abnormal pollen phenotype were found to contain a translocation when analyzed using genetic mapping. A normal pollen phenotype does not, however, guarantee the absence of a translocation. We observed one T-DNA line with normal pollen that nevertheless had a translocation based on genetic mapping results. One additional phenomenon that we observed through our genetic mapping experiments was that the T-DNA junctions on the 5'- and 3'-sides of a targeted gene can genetically separate from each other in some cases. Two of the lines in our survey displayed this 'T-DNA borders separate' phenomenon. Experimental procedures for efficiently screening T-DNA lines for the presence of chromosomal abnormalities are presented and discussed.
引用
收藏
页码:990 / 1001
页数:12
相关论文
共 32 条
[1]   DIFFERENTIAL STAINING OF ABORTED AND NONABORTED POLLEN [J].
ALEXANDER, MP .
STAIN TECHNOLOGY, 1969, 44 (03) :117-+
[2]   Genome-wide Insertional mutagenesis of Arabidopsis thaliana [J].
Alonso, JM ;
Stepanova, AN ;
Leisse, TJ ;
Kim, CJ ;
Chen, HM ;
Shinn, P ;
Stevenson, DK ;
Zimmerman, J ;
Barajas, P ;
Cheuk, R ;
Gadrinab, C ;
Heller, C ;
Jeske, A ;
Koesema, E ;
Meyers, CC ;
Parker, H ;
Prednis, L ;
Ansari, Y ;
Choy, N ;
Deen, H ;
Geralt, M ;
Hazari, N ;
Hom, E ;
Karnes, M ;
Mulholland, C ;
Ndubaku, R ;
Schmidt, I ;
Guzman, P ;
Aguilar-Henonin, L ;
Schmid, M ;
Weigel, D ;
Carter, DE ;
Marchand, T ;
Risseeuw, E ;
Brogden, D ;
Zeko, A ;
Crosby, WL ;
Berry, CC ;
Ecker, JR .
SCIENCE, 2003, 301 (5633) :653-657
[3]   T-DNA mediated disruption of essential gametophytic genes in Arabidopsis unexpectedly rare and cannot be inferred from segregation distortion alone [J].
Bonhomme, S ;
Horlow, C ;
Vezon, D ;
de Laissardière, S ;
Guyon, A ;
Férault, M ;
Marchand, M ;
Bechtold, N ;
Pelletier, G .
MOLECULAR AND GENERAL GENETICS, 1998, 260 (05) :444-452
[4]  
Budziszewski GJ, 2001, GENETICS, V159, P1765
[5]   Genetical and cytological studies of semisterility and related phenomena in maize [J].
Burnham, CR .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1930, 16 :269-277
[6]   GENETIC AND MOLECULAR CHARACTERIZATION OF EMBRYONIC MUTANTS IDENTIFIED FOLLOWING SEED TRANSFORMATION IN ARABIDOPSIS [J].
CASTLE, LA ;
ERRAMPALLI, D ;
ATHERTON, TL ;
FRANZMANN, LH ;
YOON, ES ;
MEINKE, DW .
MOLECULAR & GENERAL GENETICS, 1993, 241 (5-6) :504-514
[7]   Protocol: An improved high-throughput method for generating tissue samples in 96-well format for plant genotyping (Ice-Cap 2.0) [J].
Clark, Katie A. ;
Krysan, Patrick J. .
PLANT METHODS, 2007, 3 (1)
[8]   Reciprocal chromosome translocation associated with TDNA-insertion mutation in Arabidopsis: genetic and cytological analyses of consequences for gametophyte development and for construction of doubly mutant lines [J].
Curtis, Marc J. ;
Belcram, Katia ;
Bollmann, Stephanie R. ;
Tominey, Colin M. ;
Hoffman, Peter D. ;
Mercier, Raphael ;
Hays, John B. .
PLANTA, 2009, 229 (04) :731-745
[9]   AGROBACTERIUM-MEDIATED TRANSFORMATION OF GERMINATING-SEEDS OF ARABIDOPSIS-THALIANA - A NON-TISSUE CULTURE APPROACH [J].
FELDMANN, KA ;
MARKS, MD .
MOLECULAR AND GENERAL GENETICS, 1987, 208 (1-2) :1-9
[10]   A comprehensive characterization of single-copy T-DNA insertions in the Arabidopsis thaliana genome [J].
Forsbach, A ;
Schubert, D ;
Lechtenberg, B ;
Gils, M ;
Schmidt, R .
PLANT MOLECULAR BIOLOGY, 2003, 52 (01) :161-176