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GENETICS, GENOMICS, AND MOLECULAR EVOLUTION

Histochemical Analysis Reveals Organ-Specific Quantitative Trait Loci for Enzyme Activities in Arabidopsis¹

Laboratory of Plant Physiology (L.I.S., J.V., J.J.B.K., L.H.W.v.d.P., D.V.) and Laboratory of Genetics (J.J.B.K., M.K.), Graduate School Experimental Plant Sciences, Wageningen University, Arboretumlaan 4, 6703 BD Wageningen, The Netherlands; and Institute of Plant Physiology, Russian Academy of Sciences, Botanicheskaya 35, 127 276, Moscow, Russia (L.I.S.)

To identify genetic loci involved in the regulation of organ-specific enzyme activities, a specific histochemical staining protocol was used in combination with quantitative trait locus (QTL) analysis. Using phosphoglucomutase (PGM) as an example, it is shown that enzyme activity can specifically, and with high resolution, be visualized in non-sectioned seedlings of Arabidopsis. The intensities of staining were converted to quantitative data and used as trait for QTL analysis using Landsberg erecta x Cape Verde Islands recombinant inbred lines. Independently, PGM activities were quantified in whole-seedling extracts, and these data were also used for QTL analysis. On the basis of extract data, six significant (P < 0.05) loci affecting PGM activity were found. From the histochemical data, one or more specific QTLs were found for each organ analyzed (cotyledons, shoot apex, hypocotyl, root, root neck, root tip, and root hairs). Loci detected for PGM activity in extracts colocated with loci for histochemical staining. QTLs were found coinciding with positions of (putative) PGM genes but also at other positions, the latter ones supposedly pointing toward regulatory genes. Some of this type of loci were also organ specific. It is concluded that QTL analysis based on histochemical data is feasible and may reveal organ-specific loci involved in the regulation of metabolic pathways.

¹ This work was supported in part by the European Union (grant no. QLRT-2000 - 01097). L.I.S. was supported by the North Atlantic Treaty Organization-Linkage (grant no. CLG 978850). J.J.B.K. was supported by Netherlands Organization for Scientific Research Program Genomics (grant no. 050-10-029).

^* Corresponding author; e-mail dick.vreugdenhil{at}wur.nl; fax 31-317-484740.

Received May 27, 2003; returned for revision July 30, 2003; accepted October 9, 2003.

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