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Characterization of Grain Amino Acid Composition and Proteome Profile of a High-lysine Barley Mutant Line M98 (고-Lysine 보리 돌연변이 계통 M98 종실의 아미노산 조성 및 Proteome Profile 특성)

  • Kim, Dea-Wook;Kim, Hong-Sik;Park, Hyoung-Ho;Hwang, Jong-Jin;Kim, Sun-Lim;Lee, Jae-Eun;Jung, Gun-Ho;Hwang, Tae-Young;Kim, Jung-Tae;Kim, Si-Ju;Rakwal, Randeep;Kwon, Young-Up
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.57 no.2
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    • pp.171-181
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    • 2012
  • Lysine is the first limiting essential amino acid in cereals for humans and monogastric animals, although its content is generally low. A chemically induced high-lysine barley mutant, M98, has an agronomically undesirable shrunken endosperm trait. In order to obtain detailed insight into the atypical traits of M98 grains, we characterized amino acid composition and protein profiles of M98 and its parent cultivar Chalssalbori. Among a total of 16 amino acids, the percentage of each of the 7 amino acids, including lysine, was 1.2~1.8 times higher in M98, comparing to Chalssalbori. The percentage of proline and its precursor, glutamic acid, in M98 was about the half of that of the amino acids in Chalssalbori, but arginine synthesized from glutamic acid was 1.8 times higher in M98, compared that in the parent cultivar. Theses results indicated that the mutation in M98 grains might alter the proportion of amino acids linked to each other in a biosynthetic pathway. A comparison of grain proteome profiles between Chalssalbori and M98 revealed 70 differentially expressed protein spots, where 45 protein spots were up-regulated and 25 protein spots down-regulated in M98 compared to those in Chalssalbori. Of these changed protein spots, 53 were identified using nano-electrospray ionization liquid chromatography mass spectrometry. Most of these identified proteins were involved in various biological processes. In particular, 28 protein spots such as ${\beta}$-amylase, serpins and B3-hordein were identified as proteins associated with the atypical traits of M98. It was thought that a genetic study on the unique protein profile of M98 would be needed to develop an agronomically feasible barley cultivar with high-lysine trait.

Role of Ghrelin in the Control of Reproductive Endocrine Function (포유류 생식 내분비 기능 조절에서 Ghrelin의 역할)

  • Lee, Sung-Ho
    • Development and Reproduction
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    • v.13 no.4
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    • pp.207-215
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    • 2009
  • Numerous factors can affect the activities of hypothalamus-pituitary-gonad (HPG) hormonal axis, resulting in alteration of reproductive capacity or status such as onset of puberty and menopause. Soon after the finding of leptin, a multifunctional hormone secreted from adipocytes, a close relationship between reproduction and body energy balance have been manifested. Ghrelin, another multifunctional hormone from gastrointestinal tract, is an endogenous ligand of growth hormone secretagogue receptor (GHSR), and is thought to be a counterpart of leptin in the regulation of energy homeostasis. As expected, ghrelin can also modulate the reproductive capacity through the modulation of activities of HPG axis. This paper summarizes the current knowledge on the discovery, gene structures, tissue distribution and roles of ghrelin and GHSRs in mammalian reproduction in particular modulation of reproductive hormone secretion in HPG axis. Like POMC gene expression in pituitary gland, preproghrelin gene can generate a complex repertoire of transcripts which further undergo alternative splicing and posttranslational modifications. Concerning the roles of preproghrelin gene products in the control of body physiology except energy homeostasis, limited knowledge is available so far. Several lines of evidence, however, show the interplay of ghrelin between metabolism and reproduction. In rat and human, the distribution of ghrelin receptor GHSRs (GHSR1a and GHSR1b) has been confirmed not only in the hypothalamus and pituitary which were originally postulated as target of ghrelin but also in the testis and ovary. Expression of the preproghrelin gene in the brain and gonads was also verified, suggesting the local role (s) of ghrelin in HPG axis. Ghrelin might play a negative modulator in the secretions of hypothalamic GnRH, pituitary gonadotropins and gonadal steroids though the action on pituitary is still questionable. Recent studies suggest the involvement of ghrelin in regulation of puberty onset and possibly of menopause entry. It is now evident that ghrelin is a crucial hormomal component in 'brain-gut' axis, and is a strong candidate links between metabolism and reproduction. Opposite to that for leptin, ghrelin signaling is likely representing the 'hunger' state of body energy balance and is necessary to avoid the energy investment into reproduction which has not a top priority in maintaining homeostasis. Further researches are needed to gain a deep insight into the more precise action mechanism and role of ghrelin in reproduction, and to guarantee the successful biomedical applications.

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Effect of Moutan Cortex Radicis on gene expression profile of differentiated PC12 rat cells oxidative-stressed with hydrogen peroxide (모단피의 PC12 cell 산화억제 효과 및 neuronal 유전자 발현 profile 분석에 대한 연구)

  • Kim Hyun Hee;Rho Sam Woong;Na Youn Gin;Bae Hyun Su;Shin Min Kyu;Kim Chung Suk;Hong Moo Chang
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.529-541
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    • 2003
  • Yukmijihwang-tang has been widely used as an and-aging herbal medicine for hundred years in Asian countries. Numerous studies show that Yukmijihwangtang has anti-oxidative effect both in vivo and in vitro. It has been reported that Moutan Cortex Radicis extract (MCR) was the most effective herb in Yukmijihwang-tang on undifferentiated PC12 cells upon oxidative-stressed with hydrogen peroxide. The purpose of this study is to; 1) evaluate the recovery of neuronal damage by assessing the anti-oxidant effect of MCR on PC12 cells differentiated with nerve growth factor (NGF), 2) identify candidate genes responsible for anti-oxidative effect on differentiated PC12 cells by oligonucleotide chip microarray. PC12 cells, which were differentiated by treating with NGF, were treated without or with hydrogen peroxide in the presence or absence of various concentration of MCR. Cell survival was determined by using MTS assay. Measurement of intracellular reactive oxygen species (ROS) generation was determined using the H2DCFDA assay The viability of cells treated with MCR was significantly recovered from stressed PC12 cell. In addition, wide rage of concentrations of MCR shows dose-dependent inhibitory effect on ROS production in oxidative-stressed cells. Total RNAs of cells without treatment(Control group), only treated with H₂O₂ (stressed group) and treated with both H₂O₂ and of MCR (MCR group) were isolated, and cDNAs was synthesized using oligoT7(dT) primer. The fragmented cRNAs, synthesized from cDNAs, were applied to Affymetrix GeneChip Rat Neurobiology U34 Array. mRNA of Calcium/calmodulin-dependent protein kinase II delta subunit(CaMKII), neuron glucose transporter (GLUT3) and myelin/oligodendrocyte glycoprotein(MOG) were downregulated in Stressed group comparing to Control group. P2X2-5 receptor (P2X2R-5), P2X2-4 receptor (P2X2R-4), c-fos, 25 kDa synaptosomal attachment protein(SNAP-25a) and GLUT3 were downregulated, whereas A2 adenosine receptor (A2AR), cathechol-O-methyltransferase(COMT), glucose transporter 1 (GLUT1), EST223333, heme oxygenase (HO), VGF, UI-R-CO-ja-a-07-0-Ul.s1 and macrophage migration inhibitory factor (MIF) were upregulated in MCA group comparing to Control group. Expression of Putative potassium channel subunit protein (ACK4), P2X2A-5, P2X2A-4, Interferon-gamma inducing factor isoform alpha precursor (IL-18α), EST199031, P2XR, P2X2 purinoceptor isoform e (P2X2R-e), Precursor interleukin 18 (IL-18) were downregulated, whereas MOO, EST223333, GLUT-1, MIF, Neuronatin alpha, UI-R-C0-ja-a-07-0-Ul.s1, A2. adenosine receptor, COMT, neuron-specific enolase (NSE), HO, VGF, A rat novel protein which is expressed with nerve injury (E12625) were upregulated in MCR group comparing to Stressed group. The results suggest that decreased viability and AOS production of PC12 cell by H₂O₂ may be, at lease, mediated by impaired glucose transporter expression. It is implicated that the MCR treatment protect PC12 cell from oxidative stress via following mechanisms; improving glucose transport into the cell, enhancing expression of anti-oxidative genes and protecting from dopamine cytotoxicity by increment of COMT and MIF expression. The list of differentially expressed genes may implicate further insight on the action and mechanism behind the anti-oxidative effects of herbal extract Moutan Cortex Radicis.

Minisatellite 5 of SLC6A18 (SLC6A18-MS5): Relationship to Hypertension and Evolutional Level (SLC6A18 유전자의 minisatellites 5 (SLC6A18-MS5)의 고혈압과의 관련성 및 진화적 의미)

  • Heo, Chang-Hwan;Lee, Sang-Yeop;Seol, So-Young;Kwon, Jeong-Ah;Jeong, Yun-Hee;Chung, Chung-Nam;SunWoo, Yang-Il
    • Journal of Life Science
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    • v.18 no.12
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    • pp.1733-1738
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    • 2008
  • SLC6A18, one of the neurotransmitters, was reported the possible relationship to hypertension, and it contained eight blocks of minisatellites. In this study, SLC6A18-MS5 sequence which showed the highest heterozygosity among seven minisatellites was analyzed using the Transfac software, the putative binding sites for the transcription factor Pax4 and HNF4 were discovered as a result. The HNF4 is involved in the diabetes pathway and suggested the relationship to hypertension. Thus, we investigated the putative functional significance of allelic variation in this minisatellites with respect to susceptibility for hypertension. To address this possibility, we analyzed genomic DNA from the blood of 301 hypertension-free controls and 184 cases with hypertension. A statistically significant association was not identified between the allelic distribution of SLC6A18-MS5 and occurrence of hypertension. We then examined the meiotic segregation of SLC6A18-MS5 and it was transmitted following Mendelian inheritance. Therefore, this locus could be useful markers for paternity mapping and DNA fingerprinting. Moreover, we undertook a comprehensive analysis of the genomic sequence to address the evolutionary events of these variable repeats. SLC6A18 minisatellites regions are only conserved in human and primates. This result suggestedthat intronic minisatellites analysis is powerful evolution marker for the non-coding regions in primates and can provide a great insight to the molecular evolution of repeated region in primates.

Naringin Protects Ovalbumin-induced Asthma through the Down-regulation of MMP-9 Activity and GATA-3 Gene (Naringin에 의한 천식치료 효과연구)

  • Lee, Chang-Min;Chang, Jeong-Hyun;Jung, In-Duk;Jeong, Young-Il;Tae, Noh-Kyung;Park, Hee-Ju;Kim, Jong-Suk;Shin, Yong-Kyoo;Park, Sung-Nam;Park, Yeong-Min
    • Journal of Life Science
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    • v.19 no.6
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    • pp.735-743
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    • 2009
  • The common word flavonoids is often used to classify a family of natural compounds, highly abundant in all higher plants, that have received significant therapeutic interest in recent years. Naringin is associated with a reduced risk of heart disease, neurodegenerative disease, cancer and other chronic diseases; however the molecular basis of this effect remains to be elucidated. Thus we attempted to elucidate the anti-allergic effect of Naringin in ovalbumin (OVA)-induced asthma model mice. The OVA-induced mice showed allergic reactions in the airways. These included an increase in the number of eosinophils in bronchoalveolar lavage (BAL) fluid, an increase in inflammatory cell infiltration into the lung around blood vessels and airways, airway luminal narrowing, and the development of airway hyper-responsiveness (AHR). The administration of Naringin before the last airway OVA challenge resulted in a significant inhibition of all asthmatic reactions. Accordingly, this study may provide evidence that Naringin plays a critical role in the amelioration of the pathogenetic process of asthma in mice. These findings provide new insight into the immunopharmacological role of Naringin in terms of its effects on asthma in mice.

Construction and estimation of soil moisture site with FDR and COSMIC-ray (SM-FC) sensors for calibration/validation of satellite-based and COSMIC-ray soil moisture products in Sungkyunkwan university, South Korea (위성 토양수분 데이터 및 COSMIC-ray 데이터 보정/검증을 위한 성균관대학교 내 FDR 센서 토양수분 측정 연구(SM-FC) 및 데이터 분석)

  • Kim, Hyunglok;Sunwoo, Wooyeon;Kim, Seongkyun;Choi, Minha
    • Journal of Korea Water Resources Association
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    • v.49 no.2
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    • pp.133-144
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    • 2016
  • In this study, Frequency Domain Reflectometry (FDR) and COSMIC-ray soil moisture (SM) stations were installed at Sungkyunkwan University in Suwon, South Korea. To provide reliable information about SM, soil property test, time series analysis of measured soil moisture, and comparison of measured SM with satellite-based SM product are conducted. In 2014, six FDR stations were set up for obtaining SM. Each of the stations had four FDR sensors with soil depth from 5 cm to 40 cm at 5~10 cm different intervals. The result showed that study region had heterogeneous soil layer properties such as sand and loamy sand. The measured SM data showed strong coupling with precipitation. Furthermore, they had a high correlation coefficient and a low root mean square deviation (RMSD) as compared to the satellite-based SM products. After verifying the accuracy of the data in 2014, four FDR stations and one COSMIC-ray station were additionally installed to establish the Soil Moisture site with FDR and COSMIC-ray, called SM-FC. COSMIC-ray-based SM had a high correlation coefficient of 0.95 compared with mean SM of FDR stations. From these results, the SM-FC will give a valuable insight for researchers into investigate satellite- and model-based SM validation study in South Korea.

Functional analysis of seaR protein identified from Saccharopolyspora erythraea (희소방선균의 seaR 단백질 발현을 통한 기능 분석)

  • Ryu, Jae Ki;Kwon, Pil-Seung;Lee, Hyeong Seon
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.39-47
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    • 2015
  • Secondary metabolism in actinomycetes has been known to be controlled by a small molecule, ${\gamma}$-butyrolactone autoregulator, the binding of which to each corresponding receptor leads to the regulation of the transcriptional expression of the secondary metabolites. We expected that expression of an autoregulator receptor or a pleiotropic regulator in a non-host was to be gained insight of effective production of new metabolic materials. In order to study the function of the receptor protein (seaR), which is isolated from Saccharopolyspora erythraea, we introduced the seaR gene to Streptomyces coelicolor A3(2) as host strains. An effective transformation procedure for S. coelicolor A3(2) was established based on transconjugation by Escherichia coli ET12567/pUZ8002 with a ${\varphi}C31$-derived integration vector, pSET152, which contained int, oriT, attP and $ermEp^*$ (erythromycin promotor). Therefore, the pEV615 was introduced into S. coelicolor A3(2) by conjugation and integrated at the attB locus in the chromosome of the recipients by the ${\varphi}C31$ integrase (int) function. Exconjugant of S. coelicolor A3(2) containing the seaR gene was confirmed by PCR and transcriptional expression of the seaR gene in the transformant was analyzed by RT-PCR. In case of S. coelicolor A3(2), a phenotype microarray was used to analyze the phenotype of transformant compared with wild type by seaR expression. After that, in order to confirm the accuracy of the results obtained from the phenotype microarray, an antimicrobial susceptibility test was carried out. This test indicated that sensitivity of the transformant was higher than wild type in tetracycline case. These results indicated that some biosynthesis genes or resistance genes for tetracycline biosynthesis in transformant might be repressed by seaR expression. Therefore, subsequent experiments, analysis of transcriptional pattern of genes for tetracycline production or resistance, are needed to confirm whether biosynthesis genes or resistance genes for tetracycline are repressed or not.

Variations in Nutrients & $CO_2$ Uptake Rates of Porphyra yezoensis Ueda and a Simple Evaluation of in situ N & C Demand Rates at Aquaculture Farms in South Korea (방사무늬김(Porphyra yezoensis Ueda)의 영양염과 이산화탄소 흡수율 정밀 평가를 통한 양식해역의 질소와 탄소 요구량 산정)

  • Shim, JeongHee;Hwang, Jae Ran;Lee, Sang Yong;Kwon, Jung-No
    • Korean Journal of Environmental Biology
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    • v.32 no.4
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    • pp.297-305
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    • 2014
  • In order to understand the contribution of seaweed aquaculture to nutrients and carbon cycles in coastal environments, we measured the nutrients & carbon uptake rates of Porphyra yezoensis Ueda sampled at Nakdong-River Estuary using a chamber incubation method from November 2011 to April 2012. It was observed that the production rate of dissolved oxygen by P. yezoensis (n=30~40) was about $68.8{\pm}46.0{\mu}mol\;{g_{FW}}^{-1}h^{-1}$ and uptake rate of nitrate, phosphate and dissolved inorganic carbon (DIC) was found to be $2.5{\pm}1.8{\mu}mol\;{g_{FW}}^{-1}h^{-1}$, $0.18{\pm}0.11{\mu}mol\;{g_{FW}}^{-1}h^{-1}$ and $87.1{\pm}57.3{\mu}mol\;{g_{FW}}^{-1}h^{-1}$, respectively. There was a positive linear correlation existed between the production rate of dissolved oxygen and the consumption rates of nitrate, phosphate and DIC, respectively, suggesting that these factors may serve as good indicators of P. yezoensis photosynthesis. Further, there was a negative logarithmic relationship between fresh weight of thallus and uptake rates of nutrients and $CO_2$, which suggested that younger specimens (0.1~0.3 g) were much more efficient at nutrients and $CO_2$ uptake than old specimens. It means that the early culturing stage than harvesting season might have more possibilities to be developed chlorosis by high rates of nitrogen uptake. However, N & C demanding rates of Busan and Jeollabuk-do, calculated by monthly mass production and culturing area, were much higher than those of Jeollanam-do, the highest harvesting area in Korea. Chlorosis events at Jeollabuk-do recently might have developed by the reason that heavily culture in narrow area and insufficient nutrients in maximum yield season (Dec.~Jan.) due mostly to shortage of land discharge and weak water circulation. The annual DIC uptake by P. yezoensis in Nakdong-River Estuary was estimated about $5.6{\times}10^3\;CO_2$ ton, which was about 0.03% of annual carbon dioxide emission of Busan City. Taken together, we suggest more research would be helpful to gain deep insight to evaluate the roles of seaweed aquaculture to the coastal nutrients cycles and global carbon cycle.

Alterations of Calcium-binding Protein Immunoreactivities in the Hippocampus Following Traumatic Brain Injury (외상성 뇌손상 후 해마내 칼슘결합단백질 면역반응의 변화)

  • Oh, Yun-Jung;Kim, Baek-Seon;Park, Dae-Kyoon;Park, Kyung-Ho;Ko, Jeong-Sik;Kim, Duk-Soo
    • Applied Microscopy
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    • v.41 no.4
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    • pp.235-248
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    • 2011
  • Traumatic brain injury (TBI) is one of the leading causes of death and disability in children and adults and is a major risk factor for the development of posttraumatic epilepsy (PTE). Recent studies have provided significant insight into the pathophysiological mechanisms underlying the development of epilepsy. Although the link between brain trauma and epilepsy is well recognized, the complex biological mechanisms that result in PTE following TBI have not been fully elucidated. Therefore, this study investigated in order to identify whether or not the abnormal expression of calcium-binding proteins in the lesioned hippocampus plays a role in neuronal damage by brain trauma and whether or not the expressions may change in the contralateral hippocampus during the adaptive stage as early time point following TBI. During early time point following TBI, both parvalbumin (PV) and calbindin D-28k (CB) immunoreactivities were decreased with in the lesioned hippocampus. However, these expressions were recovered to control levels as depend on time courses. On the other hand, PV immunoreactivity in contralateral hippocampus was transiently reduced as compared to the control levels, whereas CB expression was unchanged. These findings indicate that the alterations of the calcium-binding proteins, especially PV and CB, may contribute to the neuronal death and/or damage induced by abnormal inhibitory neurotransmission at early time period following brain trauma and the development of epileptogenesis in patients with traumatic brain injury.

Growth, Photosynthesis and Chlorophyll Fluorescence of Chinese Cabbage in Response to High Temperature (고온 스트레스에 대한 배추의 생장과 광합성 및 엽록소형광 반응)

  • Oh, Soonja;Moon, Kyung Hwan;Son, In-Chang;Song, Eun Young;Moon, Young Eel;Koh, Seok Chan
    • Horticultural Science & Technology
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    • v.32 no.3
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    • pp.318-329
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    • 2014
  • In order to gain insight into the physiological responses of plants to high temperature stress, the effects of temperature on Chinese cabbage (Brassica campestris subsp. napus var. pekinensis cv. Detong) were investigated through analyses of photosynthesis and chlorophyll fluorescence under 3 different temperatures in the temperature gradient tunnel. Growth (leaf length and number of leaves) during the rosette stage was greater at ambient $+4^{\circ}C$ and ambient $+7^{\circ}C$ temperatures than at ambient temperature. Photosynthetic $CO_2$ fixation rates of Chinese cabbage grown under the different temperatures did not differ significantly. However, dark respiration rate was significantly higher in the cabbage that developed under ambient temperature relative to elevated temperature. Furthermore, elevated growth temperature increased transpiration rate and stomatal conductance resulting in an overall decrease of water use efficiency. The chlorophyll a fluorescence transient was also considerably affected by high temperature stress; the fluorescence yield $F_J$, $F_I$, and $F_P$ decreased considerably at ambient $+4^{\circ}C$ and ambient $+7^{\circ}C$ temperatures, with induction of $F_K$ and decrease of $F_V/F_O$. The values of RC/CS, ABS/CS, TRo/CS, and ETo/CS decreased considerably, while DIo/CS increased with increased growth temperature. The symptoms of soft-rot disease were observed in the inner part of the cabbage heads after 7, 9, and/or 10 weeks of cultivation at ambient $+4^{\circ}C$ and ambient $+7^{\circ}C$ temperatures, but not in the cabbage heads growing at ambient temperature. These results show that Chinese cabbage could be negatively affected by high temperature under a future climate change scenario. Therefore, to maintain the high productivity and quality of Chinese cabbage, it may be necessary to develop new high temperature tolerant cultivars or to markedly improve cropping systems. In addition, it would be possible to use the non-invasive fluorescence parameters $F_O$, $F_V/F_M$, and $F_V/F_O$, as well as $F_K$, $M_O$, $S_M$, RC/CS, ETo/CS, $PI_{abs}$, and $SFI_{abs}$ (which were selected in this study), to quantitatively determine the physiological status of plants in response to high temperature stresses.