T-DNA-associated duplication/translocations in Arabidopsis. Implications for mutant analysis and functional genomics

Frans Tax, D. M. Vernon

Research output: Contribution to journalArticle

106 Citations (Scopus)

Abstract

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 behavior - may 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.

Original languageEnglish (US)
Pages (from-to)1527-1538
Number of pages12
JournalPlant Physiology
Volume126
Issue number4
DOIs
StatePublished - 2001
Externally publishedYes

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Genomics
Arabidopsis
genomics
mutants
DNA
Chromosomes
chromosomes
Protein Kinases
Reverse Genetics
protein kinases
Light
T-DNA
receptors
Alleles
simple sequence length polymorphism
alleles
Chromosome Mapping
Genes
Embryonic Structures
genes

ASJC Scopus subject areas

  • Plant Science

Cite this

T-DNA-associated duplication/translocations in Arabidopsis. Implications for mutant analysis and functional genomics. / Tax, Frans; Vernon, D. M.

In: Plant Physiology, Vol. 126, No. 4, 2001, p. 1527-1538.

Research output: Contribution to journalArticle

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