Min-Hyuk Oh

Defects in a proteolytic step of light-harvesting complex II in anArabidopsis stay-green mutant,ore10, during dark-induced leaf senescence

During dark-induced leaf senescence (DIS), the non-functional stay-green mutantore10 showed delayed chlorophyll (Chl) degradation and increased stability in its light-harvesting complex II (LHCII). These phenomena were closely related to the formation of aggregates that mainly consisted of terminal-truncated LHCII (Oh et al., 2003). Theore10 mutant apparently lacks the protease needed to degrade the truncated LHCII. In wild-type (WT) plants, protease was found in the thylakoid fraction, but not the soluble fraction. A similar experiment using dansylated LHCII revealed that the protease degraded both WT andore10 LHCII, indicating that its stability inore10 perhaps did not result from a defect in the LHCII polypeptides themselves. Although protease activity was not present in non-senesced WT leaves, it was induced during DIS. It also was possible to diminish the high level of protease present in the thylakoids through high-salt washing, suggesting that this enzyme is extrinsically bound to the outer surface of the stroma-exposed thylakoid regions.

A Simple Method for Extraction of High Molecular Weight DNA fromPorphyra Tenera (Rhodophyta) Using Diatomaceous Earth

The innate soluble polysaccharides and phenolic compounds of marine macroalgae are serious contaminants which interfere with experimental procedures such as restriction enzyme digestion, polymerase chain reaction (PCR) and other enzymatic reactions using extracted DNA samples. The viscous polysaccharides are co-precipitated with DNA samples by isopropanol or ethanol precipitation in conventional experiment. To overcome the problem, a method for the isolation of high molecular weight DNA from Porphyra tenera is developed with the application of diatomaceous earth column. The isolated DNAs by this method were about 50-100 kb in size and could be digested well with restriction enzymes. The nuclease activity seemed to be minimal, and high reproducibility in the arbitrary primed PCR for RAPD analyses was a distinctive feature. These results suggest that this method is very efficient in isolating nucleic acid from macroalgae including Porphyra.

Effects of Six Antibiotics on the Activity of the Photosynthetic Apparatus and Ammonium Uptake of Thallus of Porphyra yezoensis

The modern integrated fish-seaweed mariculture has been tested to reduce the environmental impacts of an intensive fed culture. To obtain the best seaweed bioremediation performance, the effects of therapeutants used for fish disease control on the selected seaweed species should be considered. As a selected seaweed, Porphyra yezoensis was tested with six commercial antibiotics including erythromycin thiocyanate_A, erythromycin thiocyanate_B, oxytetracycline, doxycycline, pefloxacin, and amoxicillin trihydrate under the batch incubation at a photon flux density of 10 mol at 15. Among the tested commercial antibiotics, erythromycin thiocyanate_A, erythromycin thiocyanate_B, oxytetracycline, and doxycycline showed decreases in Fv/Fm, the photochemical efficiency of photosystem II, with a dose-dependant and time-dependant manner. From the quenching analysis of chlorophyll fluorescence, three differential patterns were observed in the antibiotics-treated Porphyra: (1) high nonphotochemical quenching (NPQ) and low photochemical quenching (qP) in the cases of Erythromycin thiocyanate_B and amoxicillin trihydrate, (2) high NPQ and high qP in the case of pefloxacin and (3) low NPQ and low qP in the case of oxytetracycline. These results indicated that antibiotics affected in various ways on the photosynthetic apparatus, reflecting differential lesion sites of antibiotics. In addition, the rates of ammonium uptake also decreased with a decrease of Fv/Fm in P. yezoensis thalli treated with erythromycin thiocyanate_B and oxytetracycline. Therefore, the four antibiotics mentioned could affect the bioremediation capacity of the selected seaweed species in the integrated fish-seaweed mariculture system due to the decrease of photosynthetic activity and the simultaneous decrease of ammonium uptake.

Effects of benzyladenine and abscisic acid on the disassembly process of photosystems in anArabidopsis delayed-senescence mutant,ore9

Using wild-type (WT) leaves and those from anore9 delayed-senescenceArabidopsis mutant, we investigated the delaying and accelerating effects of benzyladenine (BA) and abscisic acid (ABA), respectively, on the degradation process of the photosynthetic apparatus during dark-induced senescence (DIS). In the mutant, delays were seen for both the breakdown of chlorophyll (Chl) and the decrease in photochemical efficiency of photosystem II (Fv/Fm). Moreover, each step was prolonged in the disassembly process of the Chl-protein complexes. In the presence of BA, Chl degradation was retarded to a similar extent for both the mutant and the WT, but the decrease in Fv/Fm was not. However, in the presence of ABA, the two processes were accelerated in both genotypes. Therefore, although theore9 mutation causes this functional delayed-senescence, it may not be related to the non-functional delay that happens afterwards. In contrast, BA seems to affect both processes.