Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: Analysis of gene expression during potato tuber development

TLDR

The study uses a highly synchronous in vitro tuberization system and cDNA‑AFLP to target expression of patatin and ADP‑glucose pyrophosphorylase transcripts during potato tuber development. cDNA‑AFLP revealed expression kinetics matching Northern blotting, identified two lipoxygenase transcripts differentially induced at tuber formation, enabled dissection of individual transcript expression, allowed rapid band verification, and demonstrated broad applicability for detecting developmentally regulated genes.

Abstract

Summary Using a highly synchronous in vitro tuberization system, in combination with an amplified restriction fragment polymorphism (AFLP † )‐derived technique for RNA fingerprinting (cDNA‐AFLP ‡ ), transcriptional changes at and around the time point of potato tuberization have been analyzed. The targeted expression analysis of a specific transcript coding for the major potato storage protein, patatin and a second transcript, coding for ADP‐glucose pyrophosphorylase, a key gene in the starch biosynthetic pathway is described. This paper confirms that kinetics of expression revealed by cDNA‐AFLP analysis are comparable to those found in Northern analysis. Furthermore, this paper reports the isolation and analysis of two tuberspecific transcript‐derived fragments (TDFs) coding for the lipoxygenase enzyme, which are differentially induced around the time point of tuber formation. Analysis of the two lox TDFs demonstrates that it is possible to dissect the expression modalities of individual transcripts, not independently detectable by Northern analysis. Finally, it is shown that using cDNA‐AFLP, rapid and simple verification of band identity may be achieved. The results indicate that cDNA‐AFLP is a broadly applicable technology for identifying developmentally regulated genes.