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cis-Regulatory Elements for Mesophyll-Specific Gene Expression in the C₄ Plant Flaveria trinervia, the Promoter of the C₄ Phosphoenolpyruvate Carboxylase Gene

Heinrich-Heine-Universität, Institut für Entwicklungs und Molekularbiologie der Pflanzen, 40225 Düsseldorf, Germany

¹ To whom correspondence should be addressed. E-mail west{at}uni-duesseldorf.de; fax 49-211-81-14871.

C₄ photosynthesis depends on the strict compartmentalization of CO₂ assimilatory enzymes. cis-regulatory mechanisms are described that ensure mesophyll-specific expression of the gene encoding the C₄ isoform of phosphoenolpyruvate carboxylase (ppcA1) of the C₄ dicot Flaveria trinervia. To elucidate and understand the anatomy of the C₄ ppcA1 promoter, detailed promoter/reporter gene studies were performed in the closely related C₄ species F. bidentis, revealing that the C₄ promoter contains two regions, a proximal segment up to –570 and a distal part from –1566 to –2141, which are necessary but also sufficient for high mesophyll-specific expression of the ß-glucuronidase reporter gene. The distal region behaves as an enhancer-like expression module that can direct mesophyll-specific expression when inserted into the ppcA1 promoter of the C₃ plant F. pringlei. Mesophyll expression determinants were restricted to a 41-bp segment, referred to as mesophyll expression module 1 (Mem1). Evolutionary and functional studies identified the tetranucleotide sequence CACT as a key component of Mem1.

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