Understanding the Physiological and Molecular Mechanism of Persistent Organic Pollutant Uptake and Detoxification in Cucurbit Species (Zucchini and Squash)

摘要

Cucurbits pepo ssp pepo (zucchini) roots phytoextract significant amounts of persistent organic pollutants (POPs) from soil, followed by effective translocation to aboveground tissues. The closely related C. pepo ssp ovifera (squash) does not have this ability. In a DDE-contaminated field soil, zucchini roots and stems contained 3.6 and 6.6-fold greater contaminant than did squash tissues, respectively, and zucchini phytoextracted 12-times more DDE from soil than squash. In batch hydroponics, squash was significantly more sensitive to DDE (2-20 mg/ L) exposure; 4 mg/L DDE significantly reduced squash biomass (14%) whereas for zucchini, biomass reductions were observed at 20 mg/L (20%). PCR select Suppression Subtraction Hybridization was used to identify differentially expressed genes in DDE treated zucchini relative to DDE treated squash or non-treated zucchini. After differential screening to eliminate false positives, unique cDNA clones were sequenced. Out of 40 shoot cDNA sequences, 34 cDNAs have homology to parts of phloem filament protein 1 (PP1). Out of 6 cDNAs from the root tissue, two cDNAs are similar to cytochrome P450 like proteins, and one cDNA matches a putative senescence associated protein. From the DDE exposed zucchini seedlings cDNA library, out of 22 differentially expressed genes, 14 cDNAs were found to have homology with genes involved in abiotic stresses, signaling, lipid metabolism, and photosynthesis. A large number of cDNA sequences were found to encode novel unknown proteins that may be involved in uncharacterized pathways of DDE metabolism in plants. A semiquantitative RT-PCR analysis of isolated genes confirmed up-regulaton in response to DDE exposure.