Highly degenerate primers to conserved regions of the eukaryotic phosphoinositol-specific phospholipase C (PLC) were used to amplify fragments of plant PLCs from Arabidopsis thaliana genomic DNA. Eight completely different fragment sequences that showed high homology to PLCs of both animals and plants were isolated. The variation between these putative PLCs was high and suggests that, like animals, plants have multiple isoforms of PLC. Using one of the PCR clones, we isolated a corresponding full-length Arabidopsis PLC gene (ATHATPLC1G), and sequence analysis indicated that it was most like a delta-type PLC. This gene is 2.5 kb and contains seven introns, all but one of which has intron/exon border sequences that conform to the Arabidopsis consensus. The structural complexity of the gene is relatively simple compared to mammalian β-type PLCs that can be 15 kb long with up to 30 introns. The plant gene is a single copy and was mapped to four Arabidopsis YACs, one located on chromosome 2. The promoter region contained two TATA-like elements at -43 and -185 and other putative regulatory elements that suggest that this PLC is hormonally regulated. This is the first plant PLC gene and the first delta type-PLC gene from a higher organism to be sequenced.
Bibliographical noteFunding Information:
The authors wish to thank Jean Finnegan for Arabidopsis DNA and many helpful suggestions, Kathi Blomer for Arabidopsis DNA, Robin Chapple for help with YAC library screening, and Janice Norman and Jenny Thistleton for their excellent technical assistance with this project. LMH acknowledges the financial support of the Australian Cotton Research and Development Corporation.
- Degenerate PCR amplification
- Phosphoinositol-specific phospholipase C
- Plant gene regulation
- Signal transduction