Jin-Hong Kim

Alterations in the Photosynthetic Pigments and Antioxidant Machineries of Red Pepper (Capsicum annuum L.) Seedlings from Gamma-Irradiated Seeds

To characterize the stimulatory effects of low-dose gamma radiation on early plant growth, we investigated alterations in the photosynthesis and antioxidant capacity of red pepper (Capsicum annuum L.) seedlings produced from gamma-irradiated seeds. For two cultivars (Yeomyung and Joheung), three irradiation groups (2, 4, and 8 Gy, but not 16 Gy) showed enhanced development, although Fv/Fm, the maximum photochemical efficiency of Photosystem II (PSII), did not differ significantly among any of the four groups. In contrast, values for 1/Fo — 1/Fm, i.e., a measure of functional PSII content, decreased in the irradiated groups of ‘Yeomyung’ but increased in those of ‘Joheung’. Pigment analyses and enzyme activity assays revealed that irradiation altered the compositions of photosynthetic pigments (chlorophylls and carotenoids) as well as the activities of antioxidant enzymes (superoxide dismutase and glutathione reductase). However, these shifts were not directly related to the increase in early growth, although they were cultivar-and developmental stage-dependent In addition, the effects of irradiation on the enzymatic activities measured here were at opposition between the two cultivars.

Effects of in Planta Gamma-Irradiation on Growth, Photosynthesis, and Antioxidative Capacity of Red Pepper (Capsicum annuum L.) Plants

We investigated the effects of low-dose inplanta irradiation on red pepper plants treated with gamma rays of 2, 4, 8, and 16 Gy. Growth was stimulated at 2 and 4 Gy but inhibited at 8 and 16 Gy. Photochemical quenching (qP) increased slightly in all treatment groups for 1 d after irradiation (DAl), whereas non-photochemical quenching (NPQ) decreased more noticeably. These changes in qP and NPQ were transient and had almost recovered to the control level by 2 DAl. Although carotenoid pigments also fluctuated during the experimental period, chlorophylls were almost entirely insensitive to the gamma rays. Irradiation also partially protected leaves from a decrease in photochemical efficiency (Fv/Fm) under conditions of UV-B (2.2 W m-2) and high light intensity (800 μmol m-2 s-1). This enhanced stress resistance could be partly explained by higher levels of SOD and APX activities, as well as ascorbate content. Our results demonstrate for the first time that the carotenoid pigments are the most radio-sensitive and fastest recovering compounds in plants, and that SOD, APX, and ascorbate are important inducible factors for improving stress resistance through the use ofin planta gamma-irradiation.

Ultrastructural changes of cell organelles inArabidopsis stems after gamma irradation

We examined ultrastructural changes of the cell organelles ofArabidopsis stems in response to gamma irradiation. Seedlings treated with 0 to 5 Gy developed normally, while height growth in plants exposed to 50 Gy was significantly inhibited. Based on TEM observations, the chloroplasts were extremely sensitive to such irradiation. In particular, the thylakoids were heavily swollen, some portions of the mitochondria and endoplasmic reticulum were structurally altered, and the plasmalemma had pulled away from the cell wall in places. However, no ultrastructural changes in cell organelles occurred at doses of 0 to 5 Gy.

cDNA cloning and sequence analysis of the rice Cinnamate-4-Hydroxylase gene, a cytochrome P450-dependent monooxygenase involved in the general phenylpropanoid pathway

Plant cytochrome P450 monooxygenases (P450s) mediate a wide range of oxidative reactions involved in the biosynthesis of phenylpropanoids, terpenes, lipids, and alkaloids. We isolated a cDNA clone for cinnamate-4-hydroxylase (C4H) from a Japonica type rice(Oryza sativa L. cv. llpumbyeo). This C4H has a deduced amino acid sequence that is 85% identical tothat ofSorghum bicolor. Our phylogenetic analysis also showed that theOsC4HL gene is closely related toC4H fromS. bicolor. A putative genomic DNA sequence corresponding toOsC4HL contained cis-elements (boxes P, A, L, and TCA motifs), AT-rich elements, and wound-response elements that control gene expression in its promoter region.OsC4HL expression was detected in all the tissue types, with the highest level being measured in the roots. It was also apparently up-regulated by wounding stress. These data suggest that theOsC4HL gene isC4H member in theCYP73 subfamily.

Localization of hydrogen peroxide in pumpkin (Cucurbita ficifolia bouché) seedlings exposed to high-dose gamma ray

Hydrogen peroxide (H2O2) was detected cytochemically, via transmission electron microscopy (TEM), in pumpkin tissues exposed to high-dose gamma ray. Its reaction with cerium chloride produced electron-dense precipitates of cerium perhydroxides. Their patterns of deposition in the tissues of both control plants and those irradiated with gamma ray (PIG) were typically found in the plasma membranes and cell walls. However, gamma irradiation remarkably increased the intensities of cerium perhydroxide deposits (CPDs) in the plasma membranes and cell walls for all tissue types, but especially the leaves. The only exception was for vessels in the cotyledons. After gamma irradiation, the (H2O2) content in all tissues was higher than in the control samples, except for the cotyledons of PIG, where the (H2O2) content was lower than for all others. The increased appearance of CPDs may have been due to the enhancement of (H2O2) accumulation by gamma radiation. This accumulation also varied according to the cell or tissue type examined.

Cloning, characterization, and expression of two cDNA clones for a rice ferulate-5-hydroxylase gene, a cytochrome P450-dependent monooxygenase

Ferulate-5-hydroxylase (F5H) is a cytochrome P450-dependent monooxygenase that catalyses the hydroxylation of ferulic acid, coniferaldehyde, and coniferyl alcohol in the pathways leading to sinapic acid and the syringyl unit of lignin. We have isolated twoF5H genes,OsF5HL andOsF5HL2, from a japonica-type rice (Oryza sativa L. cv. llpoombyeo). They are the firstF5H genes experimentally identified in monocotyledonous plants. Phylogenetic analysis indicated that both genes are closely related to dicotF5H genes fromArabidopsis thaliana andBrassica napus. Southern blot analysis showed that these genes exist as single copies in the rice genome. Alignments of theOsF5HL andOsF5HL2 cDNAs to their genomic DNAs revealed thatOsF5HL has an open reading frame (ORF) of 1590 b from four exons, whileOsF5HL2 has an ORF of 1560 b from two exons. Expression ofOsFSHL is highest in young leaves, whereas that ofOsF5HL2 is greatest in mature leaves. In the roots and stems, transcription levels for both genes are markedly low. These data suggest that theOsF5HL andOsF5HL2 genes belong to theCYP84 subfamily and that their expressions are tissue-specific.

Gamma radiation and hormone treatment as tools to reduce salt stress in rice (Oryza sativa L.)

We investigated the effects of jasmonic acid (JA) and gamma irradiation on the growth and metabolic responses to salt stress in rice (Oryza sativa L.) plants. The relative growth rate (RGR), relative water content (RWC), and chlorophyll (Chl) content were lower in NaCI-treated plants than in the control, whereas the malondialdehyde content (MDA), electrolyte leakage (EL), and contents of proline and abscisic acid (ABA) were higher in the treated plants. When induced by the salt stress, those effects, however, were somewhat alleviated by the application of JA or gamma irradiation. The most significant response was manifested by the proline content, with relatively lower values for alleviation being recorded for the contents of RGR, RWC, Chl, and MDA, as well as EL. Moreover, although total Chl content was not significantly influenced by JA or gamma irradiation in plants under salt stress, an increase in the level of Chl a resulted in a markedly changed ratio of Chl a/b. The degree of alleviation, in terms of growth and metabolic responses, was more extensive for JA-treated plants than for those exposed to gamma irradiation.

Alleviation of Low-temperature Photoinhibition in Gamma-irradiated Red Pepper (Capsicum annuum L.) Plants

We studied the radiation-induced stress resistance in red pepper leaves under conditions of low-temperature photoinhibition or artificially induced photo-oxidative stress. Plants irradiated with 4, 8, or 16-Gy gamma rays were more resistant to both stress factors than were the controls. However, exposure to a low temperature for 12 h with illumination or photo-oxidative treatment for 1 h differentially affected the irradiated leaves, although they had similar stress intensities as defined by their maximal photochemical efficiencies (Fv/Fm). Decreases in Fv/Fm induced by the two stress factors were instead alleviated, dose-dependently, by as much as 22 to 41% (low temperature) or 14 to 29% (photo-oxidation) in the irradiated groups. In contrast, non-photochemical quenching (NPQ) and the de-epoxidation state of xanthophyll cycle pigments could not be correlated with this enhanced stress resistance in the irradiated groups. These results suggest that the adaptive response of plants exposed to gamma radiation is more effective in protecting against low-temperature photoinhibition than against photo-oxidative stress. We also discuss here the involvement of antioxidative defense systems for increased resistance against low-temperature photoinhibition in irradiated red pepper.