• KOREAN JOURNAL OF CROP SCIENCE
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• v.47 no.5
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• pp.341-351
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• 2002

In order to examine the mechanistic basis for differential sensitivities to chilling and subsequent recovery between two rice (Oryza sativa L.) cutivars, a chilling-tolerant japonica type (Ilpumbyeo) and a chilling-susceptible indica type (Taebaekbyeo), changes of physiological responses and antioxidant enzymes were investigated. Both cultivars at 3 leaf stage were exposed at a low temperature of $5^{\circ}C$ for 3 days and subsequently recovered in a growth chamber at a $25^{\circ}C$ for 5 days with 250 mmol $m^{-2}$ $s^{-1}$. Physiological parameters such as leaf fresh weight, relative water content, cellular leakage, lipid peroxidation, and chlorophyll a fluorescence showed that the chilling tolerant cultivar had a high tolerance during chilling. However, the chilling-susceptible cultivar revealed severe chilling damages. The chilling-tolerant cultivar was also faster in recovery than the chilling-susceptible cultivar in all parameters examined. We analyzed the activity and isozyme profiles of four antioxidant enzymes which are: superoxide dismutase (SOD), caltalase (CAT), ascorbate peroxidase (APX), and glutation reductase (GR). We observed that chilling-tolerance was due to a result of the induced or higher antioxidant enzyme system, CAT and APX in leaves and SOD, CAT, APX, and GR in roots. Especially, we observed the most significant differences between the chilling-tolerant cultivar and -susceptible cultivar in CAT and APX activity. Also in isozyme profiles, CAT and APX band intensity in the chilling-tolerant cultivar was distinctively higher than in the chilling-susceptible cultivars during chilling and recovery. Thus, the cold stability of CAT and APX are expected to contribute to a tolerance mechanism of chilling in rice plants. In addition, the antioxidative enzymes activity in roots may be more important than in that of leaves to protect chilling damage on rice plants.

• Journal of Life Science
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• v.19 no.4
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• pp.442-448
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• 2009

We investigated photooxidation and responses of antioxidant enzymes involved in scavenging reactive oxygen species (ROS) after light-chilling ($4^{\circ}C$) for 2 days and post chilling ($25^{\circ}C$) in rice leaves. Chilling leaves indicated a 50% reduction in photosynthetic efficiency ($F_v/F_m$ ratio) and a 48% increase of $H_2O_2$, respectively, compared to the control group. In comparison with the control, activities of superoxide dismutase (SOD) and glutathione reductase (GR) increased at light-chilling and post-chilling. CuZn-SOD and Mn-SOD among SOD forms were detected in rice leaves, while Fe-SOD was not found. The increase of SOD and GR activity may serve as a basis for defense against chilling injury as it dismutase superoxide generated by light-chilling. Catalase (CAT) activity decreased during light-chilling, while activity of APX showed remarkable increase during light-chilling in rice leaves. Among CAT isoforms analyzed by 10% native PAGE, activities of isoform -2 and -3 were inhibited during light-chilling. From the elevated APX activity and decreased CAT activity, we suggest that these two enzymes show mutual supplementary relationships, indicating different tendency during light-chilling.

• Journal of Bio-Environment Control
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• v.16 no.4
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• pp.303-308
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• 2007

An accumulation levels of osmolytes in chilling-tolerant and chilling-sensitive cultivar of Cucurbits against chilling stress were determined during chilling stress. Total soluble sugar contents in tolerant cultivar did not changes fur 10 days after chilling stress, but then slightly increased 20 days after chilling stress. In sensitive cultivar, it was increased rapidly in the beginning of chilling stress, and increased 3.4 times as much 20 days after chilling stress as compared with unstressed plants. Proline contents in tolerant cultivar was rapidly increased by the beginning of chilling stress, and then increased 26.6 times 20 days after chilling stress as compared with unstressed plants. In sensitive cultivar, it was increased 22.0 times 20 days after chilling stress as compared with unstressed plants. A levels of glycine betaine (GB) in tolerant cultivar increased 1.9 times as much during the 20 days of chilling stress. However, concentration of GB in sensitive cultivar did not change during the chilling stress. When plants were treated exogenous GB as a foliar spray, chilling tolerance was significantly enhanced in both cultivars. The foliar application of exogenous GB was induced chilling tolerance by accumulation of GB in the plant organs. However, it does not accumulate endogenous proline.

• Journal of Bio-Environment Control
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• v.16 no.1
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• pp.54-61
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• 2007

To determine whether antioxidant enzyme systems are related to chilling tolerance, changes of antioxidant enzyme activities during the chilling stress were determined in the leaves of a chilling-tolerant cultivar (Cucurbita ficifolia, cv. Heukjong) and a chilling-sensitive cultivar (Cucurbita moschata, cv. Jaerae 13). Leaves of chilling-tolerant plant have two major isoforms, Fe-SOD and Mn-SOD, at the Rm values of 0.20 and 0.52, respectively. In leaves of chilling-sensitive plant, two major isozymes of SOD was observed, one isoform is Mn-SOD at the Rm value of 0.20, and the other isoform is Cu/zn-SOD at the nm value of 0.58. When plants were treated with chilling stress, Cu/zn-SOD at the Rm value of 0.58 was newly expressed at 10 days after chilling stress in the chilling-tolerant plants, and density of this band increased at five days after chilling stress in the chilling-sensitive plants. One APX isozyme band was observed in unstressed plants of both cultivars. Under the chilling stress one APX isozyme band was newly expressed at 10 days after chilling stress in the chilling-tolerant cultivar. Significant genotype differences were observed fnr POD isozyme banding patterns such as few main isozyme bands in chilling-tolerant plants, and one band in chilling-sensitive plants. Densities of three POD isozyme bands at the Rm of 0.36, 0.40 and 0.54 increased at 10 days after chilling stress in the chilling-tolerant plants, while two bands at the nm of 0.36 and 0.54 increased at 10 days and 20 days after chilling stress in the chilling-sensitive plants, respectively. Activities of SOD, APX and POD significantly increased during five days after chilling stress in both cultivars. In the chilling-tolerant cultivar, activities of these enzymes were higher in chilling-stressed plant than in unstressed plants. However, activities of these enzymes in the chilling-sensitive cultivar decreased rapidly after five days of chilling stress, and were lower in chilling stressed plants than in unstressed plants.

• Journal of Photoscience
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• v.3 no.1
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• pp.15-21
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• 1996

To investigate the chilling sensitivity related injuries in the photosynthetic apparatus of cucumber leaves, the light-chilling induced alterations of chlorophyll fluorescence transients in cucumber leaves were compared with those in pea leaves. As an early effect of light-chilling, an increase in Fp/Fm$^*$ was observed in both pea and cucumber leaves, which was saturated by about 6 h chilling. However, the saturated value of Fp/Fm was almost 1.0 in cucumber, in contrast to about 0.8 in pea. During the recovery period after 24 h chilling, the light-chilling induced changes in pea seemed to be reversed, but those in cucumber leaves were thought to be irreversible, because Fo was increased significantly. Light-chilling caused significant decreases in qQ and qE in cucumber leaves, but qR was increased until 6 h, and decreased thereafter. In both pea and cucumber leaves, Fm was increased by 2 h dark treatment. The Fm from the predarkened pea leaf discs was higher than the value from the preilluminated ones during the whole period of light-chilling (500 $\mu$mol m$^{-2}$s$^{-1}$ PAR). However, the predarkened cucumber leaf discs showed a reduction in Fm and an increase in Fo during the 2 h chilling in the light. These results indicate that the causes of chilling sensitivities in photosynthetic apparatus of cucumber leaves are possibly related with the damage in PSI reaction center and the ability of acidification of lumen by PSII.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.44 no.2
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• pp.171-175
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• 1999

Morphological alteration of floral organ development in rice affected by chilling stress was examined. Three varieties of rice were grown under natural conditions and subjected to 12$^{\circ}C$ for 3 or 6 days starting from the ineffective tillering stage, before heading stage and returned to natural condition. Headings were delayed by a 6 day chilling treatment. After heading the panicles were collected and examined for any possible alteration in floral organ development. It appears that there were some differences in sensitivity to chilling stress and degree of injury depending on treatment stages and variety. Chuchungbyeo was the most frequent in producing abnormal flowers among the three varieties examined. Meiosis stage was shown to be the most vulnerable to chilling stress in both Chuchungbyeo and Ilpumbyeo and young panicle differentiation stage was the frequent stage to alter flower development in response to chilling stress only in Chuchungbyeo. It was confirmed that abnormalities occurred in pollen due to chilling stress is a major factor leading to low yield, but to some extent the alterations in carpel development may playa certain role in determining a total yield in response to chilling stress at the reproduction stage in rice. There were abnormalities like extra stigmata, extra lemma, double ovary as well as abnormal anther formation in response to chilling stress. Further studies of the phenocopy observed in rice floral development may be useful for an understanding of the resistance against chilling injury during reproductive stages in rice.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.37 no.3
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• pp.288-298
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• 1992

Since the major important factors limiting plant growth and crop productivity are environmental stresses, of which low temperature is the most serious. It has been well known that many physiological processes are alterant in response to the environmental stress. With regard to the relationship between plant hormones and the regulation of chilling tolerance in rice seedlings, the major physiological roles of plant hormones: abscisic acid, ethylene and polyamines are evaluated and discussed in this paper. Rice seedlings were grown in culture solution to examine the effect of such plant hormones on physiological characters related to chilling tolerance and also to compare the different responses among tested cultivars. Intact seedlings about 14 day-old were chilled at conditions of 5$^{\circ}C$ and 80% relative humidity for various period. Cis-(＋)-ABA content was measured by the indirect ELISA technique. Polyamine content and ethylene production in leaves were determined by means of HPLC and GC respectively. Chilling damage of seedlings was evaluated by electrolyte leakage, TTC viability assay or servival test. Our experiment results described here demonstrated the physiological functions of ABA, ethylene, and polyamines related to the regulation of chilling tolerance in rice seedlings. Levels of cis-(＋)-ABA in leaves or xylem sap of rice seedlings increased rapidly in response to 5$^{\circ}C$ treatment. The tolerant cultivars had significant higher level of endogenous ABA than the sensitive ones. The ($\pm$)-ABA pretreatment for 48 h increased the chilling tolerance of the sensitive indica cultivar. One possible function of abscisic acid is the adjustment of plants to avoid chilling-induced water stress. Accumulation of proline and other compatible solutes is assumed to be another factor in the prevention of chilling injuies by abscisic acid. In addition, the expression of ABA-responsive gene is reported in some plants and may be involving in the acclimation to low temperature. Ethylene and its immediate precusor, 1-amincyclopropane-1-carboxylic acid(ACC) increased significantly after 5$^{\circ}C$ treatment. The activity of ACC synthase which converts S-adenosylmethionine (SAM) to ACC enhanced earlier than the increase of ethylene and ACC. Low temperature increased ACC synthase activity, whereas prolonged chilling treatment damaged the conversion of ACC to ethylene. It was shown that application of Ethphon was beneficial to recovering from chilling injury in rice seedlings. However, the physiological functions of chilling-induced ethylene are still unclear. Polyamines are thought to be a potential plant hormone and may be involving in the regulation of chilling response. Results indicated that chilling treatment induced a remarkable increase of polyamines, especially putrescine content in rice seedlings. The relative higher putrescine content was found in chilling-tolerant cultivar and the maximal level of enhanced putrescine in shoot of chilling cultivar(TNG. 67) was about 8 folds of controls at two days after chilling. The accumulation of polyamines may protect membrane structure or buffer ionic imbalance from chilling damage. Stress physiology is a rapidly expanding field. Plant growth regulators that improve tolerance to low temperature may affect stress protein production. The molecular or gene approaches will help us to elucidate the functions of plant hormones related to the regulation of chilling tolerance in plants in the near future.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.52 no.3
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• pp.264-273
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• 2007

In order to examine the cross-tolerance of two chilling-tolerant cultivars (Donganbyeo and Heukhyangbyeo) and two chilling-susceptible cultivars (Hyangmibyeo and Taekbaekbyeo) to salt, paraquat, and drought, changes of physiological response and antioxidant enzymes were investigated. The seedlings were grown in a growth chamber until the 4-leaf stage. The seedlings were exposed to chilling at $5^{\circ}C$ for 3 days. For drought treatment, the seedlings were subjected to drought by withholding water from plants for 5 days. For paraquat study, plants were sprayed with $300{\mu}M$ paraquat. For the salt stress, the seedlings were transferred to the Hoagland's nutrient solution containing 0.6% (w/v) NaCl for 4 days. Chilling-tolerant cultivars showed cross-tolerant to other stresses, salt, paraquat, and drought in physiological parameters, such as leaf injury, chlorophyll a fluorescence, and lipid peroxidation. The baseline levels of antioxidative enzyme activities, catalase (CAT) and peroxidase (POX) activities in chilling-tolerant cultivars were higher than in the chilling-susceptible cultivars. However, there were no differences in ascorbate peroxidase (APX) and glutathione reductase (GR) activities between chilling-tolerant and -susceptible cultivars in untreated control. CAT activity in chilling-tolerant cultivars was higher than that in chilling-susceptible cultivars during chilling, salt, and drought treatments, but not during paraquat treatment. However, other antioxidative enzymes, APX, POX, and GR activities showed no significant differences between chilling-tolerant and -susceptible cultivars during chilling, salt, paraquat, and drought treatments. Thus, it was assumed that CAT contribute to cross-tolerance mechanism of chilling, salt, and drought in rice plants.

• Journal of Environmental Science International
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• v.11 no.11
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• pp.1173-1181
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• 2002

The present study was undertaken to investigate the effect of low temperature and salicylic acid(SA) on the chilling tolerance of acclimated and nonacclimated cucumber(Cucurmis sativus L.) seedlings. The acclimation phenomenon was characterized in chilling-sensitive cucumber seedlings and found to have a significant effect on the survival and shoot dry weights. The injuries experienced by the acclimated seedlings in the third leaf stage were on average smaller by half than those experienced by the nonacclimated seedlings. Chilling also caused a large increase in the free proline levels, regardless of the acclimation status. Exogenous treatment with SA(0.5mM) resulted in improved growth and survival of the nonacclimated chilled seedlings, indicating that SA induced chilling tolerance and SA and acclimation had common effects. The application of cycloheximide in the presence of SA restored the acclimation-induced chilling tolerance. The elevated proline level observed in the cold-treated and SA-treated plants was more pronounced in the light than in the dark at a chilled temperature, indicating that endogenous proline may play a role in chilling tolerance by stabilizing the water status in response to chilling. From these results it is suggested that SA provided protection against low-temperature stress by increasing the proline accumulation, and pre-treatment with SA may induce antioxidant enzymes leading to increased chilling tolerance.

• Journal of Photoscience
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• v.9 no.2
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• pp.59-62
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• 2002

In chilling-sensitive plants, the donor side of Photosystem II is inhibited by the chilling treatment in the dark, while the acceptor side of Photosystem I is inhibited by the chilling under the moderate light. Since the addition of inhibitors of electron transfer from Photosystem II protects Photosystem I from chilling induced photoinhibition of Photosystem I, inhibition or down-regulation of Photosystem II activity in vivo may also protect Photosystem I from photoinhibition. It was revealed that dark-chilling pretreatment actually protected Photosystem I from photoinhibition. The results imply that down-regulation of Photosystem II under stress conditions may have a role to protect Photosystem I from photoinhibition.