| Abstract Detail
Pepper - Afternoon Lee, Je Min [1], Lee, Sang-Jik [2], Baek, Hyang-Ku [1], Rose, Jocelyn [3], Kim, Byung-Dong [1]. Secretome mining using yeast-based signal sequence trap in pepper fruit. TO identify extracellular proteins that may play a role in fruit development and ripening, the yeast-based signal sequence trap (YSST) screening method was carried out using a pepper fruit cDNA library. A total of 80 genes that are predicted to encode both wall-localized or plasma membrane-bound proteins were identified, some of which have not been described previously. Subcellular localization of these identified proteins has been studied using transient GFP-fusion assay and a newly developed in planta signal sequence trap method. These approaches enabled the confirmation of the extracellular localization of the secretome. In addition, RNA gel blot analyses were used to provide further insights into their expression and regulation during fruit development and ripening of wild-type and Soft flesh fruit mutant. One of the secretome tested, CaPG1,encoding a polygalacturonase gene, differed strikingly in its expression patterns between wild type and Soft flesh fruit. Sequence analysis revealed that the CaPG1 of wild type contains a single G to A transition at the 3' splice junction of intron Ⅷ resulting in cryptic splicing site and putative premature translational termination. This point mutation showed co-segregation with the Soft flesh phenotype in segregation F2 population. Integrating our data, we conclude that CaPG1 is a critical contributor to pepper fruit softening, indicating that the YSST provides a valuable functional genomics tool to identify substantial numbers of novel extracellular proteins that are associated with specific biological processes.
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1 - Seoul National University, Department of Plant Science, Center for Plant Molecular Genetics and Breeding Research, Seoul, 151-921, Korea
2 - Cornell University, Department of Plant Biology, Center for Plant Molecular Genetics and Breeding Research, Ithaca, NY, 14853, USA
3 - Cornell University, Plant Biology, 228 plant sciences, Ithaca, NY, 14853, USA
Keywords:
Capsicum
secretome
yeast-based signal sequence trap
Soft flesh
polygalacturonase.
Session: SAT08-9
Location: Hall of Ideas Room H/Monona Terrace
Date: Wednesday, July 26th, 2006
Time: 5:00 PM
Abstract ID:389 |
