Microarray-based rapid cloning of an ion accumulation deletion mutant in Arabidopsis thaliana.
Here we describe the development of a microarray-based mapping strategy to rapidly isolate deletion mutant genes. The presented approach is particularly useful for mapping mutant genes that are difficult to phenotype. This strategy uses masking bulk segregant analysis to mask unrelated deletions, thus allowing identification of target deletions by microarray hybridization of pooled genomic DNA from both WT and mutant F2 populations. Elemental profiling has proven to be a powerful tool for isolation of nutrient and toxic metal accumulation mutants in Arabidopsis. Using microarray mapping, a sodium overaccumulation mutant FN1148 was identified as having a 523-bp genomic deletion within the second exon and intron of the AtHKT1 gene. Further cosegregation, complementation, and comparative analyses among different salt-sensitive mutants confirmed that the deletion within the AtHKT1 gene is responsible for the sodium overaccumulation in shoots and leaf sodium sensitivity of the FN1148 mutant. These results demonstrate that microarray-based cloning is an efficient and powerful tool to rapidly clone ion accumulation or other genetic deletion mutants that are otherwise difficult to phenotype for mapping, such as metabolic or cell signaling mutants.
Gong
Ji-Ming
JM
Division of Biological Sciences, Cell and Developmental Biology Section and Center for Molecular Genetics, University of California at San Diego, La Jolla, CA 92093-0116, USA.
Waner
David A
DA
Horie
Tomoaki
T
Li
Shi Lun
SL
Horie
Rie
R
Abid
Khush B
KB
Schroeder
Julian I
JI
D017360:Arabidopsis / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods*; D005816:Genetic Complementation Test; D007477:Ions; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D030821:Plants, Genetically Modified; D017384:Sequence Deletion; D012964:Sodium / Q000378:metabolism; D016339:Spectrometry, Mass, Fast Atom Bombardment
D007477:Ions; D012964:Sodium