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Plant Physiol, August 2001, Vol. 126, pp. 1527-1538
T-DNA-Associated Duplication/Translocations in Arabidopsis.
Implications for Mutant Analysis and Functional
Genomics1
Frans E.
Tax2 and
Daniel M.
Vernon2*
Department of Molecular and Cellular Biology, University of
Arizona, Tucson, Arizona 85721 (F.E.T.); and Biology Department,
Whitman College, Walla Walla, Washington 99362 (D.M.V.)
T-DNA insertion mutants have become a valuable resource for studies
of gene function in Arabidopsis. In the course of both forward and
reverse genetic projects, we have identified novel interchromosomal
rearrangements in two Arabidopsis T-DNA insertion lines. Both
rearrangements were unilateral translocations associated with the left
borders of T-DNA inserts that exhibited normal Mendelian segregation.
In one study, we characterized the embryo-defective88 mutation. Although emb88 had been mapped to chromosome
I, molecular analysis of DNA adjacent to the T-DNA left border revealed
sequence from chromosome V. Simple sequence length polymorphism mapping of the T-DNA insertion demonstrated that a >40-kbp region of
chromosome V had inserted with the T-DNA into the emb88
locus on chromosome I. A similar scenario was observed with a
prospective T-DNA knockout allele of the LIGHT-REGULATED
RECEPTOR PROTEIN KINASE (LRRPK) gene.
Whereas wild-type LRRPK is on lower chromosome IV,
mapping of the T-DNA localized the disrupted LRRPK
allele to chromosome V. In both these cases, the sequence of a single
T-DNA-flanking region did not provide an accurate picture of DNA
disruption because flanking sequences had duplicated and inserted, with
the T-DNA, into other chromosomal locations. Our results indicate that
T-DNA insertion lines even those that exhibit straightforward genetic behaviormay contain an unexpectedly high frequency of rearrangements. Such duplication/translocations can interfere with reverse genetic analyses and provide misleading information about the molecular basis
of mutant phenotypes. Simple mapping and polymerase chain reaction
methods for detecting such rearrangements should be included as a
standard step in T-DNA mutant analysis.
1
This work was supported by the National Science
Foundation (award IBN-960-4344 to D.M.V.) and by the U.S. Department
of Agriculture (award no. 97353044708 to F.E.T.).
2
These authors contributed equally to the paper.
*
Corresponding author; e-mail vernondm{at}whitman.edu; fax
509-527-5904.
© 2001 American Society of Plant Physiologists
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