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1.
Acquired thermotolerance is a complex physiological phenomenon that enables plants to survive normally lethal temperatures. This study characterizes the temperature sensitivity of Arabidopsis using a chlorophyll accumulation bioassay, describes a procedure for selection of acquired thermotolerance mutants, and provides the physiological characterization of one mutant (AtTS02) isolated by this procedure. Exposure of etiolated Arabidopsis seedlings to 48 degrees C or 50 degrees C for 30 min blocks subsequent chlorophyll accumulation and is eventually lethal. Arabidopsis seedlings can be protected against the effects of a 50 degrees C, 30-min challenge by a 4-h pre-incubation at 38 degrees C. By the use of the milder challenge, 44 degrees C for 30 min, and protective pretreatment, mutants lacking components of the acquired thermotolerance system were isolated. Putative mutants isolated by this procedure exhibited chlorophyll accumulation levels (our measure of acquired thermotolerance) ranging from 10% to 98% of control seedling levels following pre-incubation at 38 degrees C and challenge at 50 degrees C. The induction temperatures for maximum acquired thermotolerance prior to a high temperature challenge were the same in AtTS02 and RLD seedlings, although the absolute level of chlorophyll accumulation was reduced in the mutant. Genetic analysis showed that the loss of acquired thermotolerance in AtTS02 was a recessive trait. The pattern of proteins synthesized at 25 degrees C and 38 degrees C in the RLD and AtTS02 revealed the reduction in the level of a 27-kD heat shock protein in AtTS02. Genetic analysis showed that the reduction of this protein level was correlated with the acquired thermotolerance phenotype. 相似文献
2.
High temperature effects on photosynthetic activity of two tomato cultivars with different heat susceptibility 总被引:13,自引:0,他引:13
Camejo D Rodríguez P Morales MA Dell'Amico JM Torrecillas A Alarcón JJ 《Journal of plant physiology》2005,162(3):3065-289
The functional activities of the photosynthetic apparatus of two tomato cultivars of different thermotolerance were investigated after a short period of high temperature treatment. Seedlings of two tomato genotypes, Lycopersicon esculentum var. Campbell-28 and the wild thermotolerant Nagcarlang, were grown under a photoperiod of 16h at 25 degrees C and dark period of 8h at 20 degrees C. At the fourth true leaf stage, a group of plants was exposed to heat stress of 45 degrees C for 2 h. The heat shock treatment caused important reductions of the net photosynthetic rate (Pn) of Campbell-28 plants due to non-stomatal components. These non-stomatal effects were not evident in Nagcarlang-treated plants. This reduction in the CO2 assimilation rate observed in Campbell-28 was generated by affections in the Calvin cycle and also in the PSII functioning. No changes in these parameters were observed in the thermotolerant genotype after the stress. Injury to the plasma membrane because of the heat stress was evident only in the Campbell-28 genotype. Heat led to a sun-type adaptation response of the photosynthesis pigment apparatus for the Nagcarlang genotype, but not for Campbell-28, and thus an increase in chlorophyll a/b ratio and a decrease in chlorophyll/carotenoid ratio were shown in Nagcarlang stressed plants. 相似文献
3.
Photosynthetic Decline from High Temperature Stress during Maturation of Wheat : II. Interaction with Source and Sink Processes
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High temperature stress reduces grain growth in wheat (Triticum aestivum L.) by altering source activity and sink capacity. The impact of stress on source and sink interactions in two wheat cultivars of differing source thermotolerance was monitored by analysis of chlorophyll fluorescence transients, Fv (variable fluorescence) and PSM (peak, stationary, maximum), of attached flag leaves on intact and decapitated tillers grown at optimum (20°C) and stress (35°C) temperatures after anthesis. The thermotolerant cultivar Waverly had reduced Fv and PS quenching and a large increase of SM during heat stress. The less thermotolerant cultivar, Len, exhibited increased Fv and PS quenching and a small increase of SM. Fluorescence induction was similar in intact and decapitated tillers of Len, indicating diminished sinksource interaction during heat stress. The present results and previous observations of photosynthetic activities indicate that cyclic electron transport and photophosphorylation in flag leaves of the thermotolerant cultivar were stimulated by sink demand (increased SM in intact plants). Reduced grain development in the thermolabile cultivar resulted from limited capacity to support cyclic electron transport and photophosphorylation (slight increase in SM of intact plants and large reduction of Cytochrome f/b6-mediated electron transport capacity). It was concluded that heat stress injures the photosynthetic apparatus during reproductive growth of wheat and that diminished source activity and sink capacity may be equally important in reducing productivity. 相似文献
4.
Chinese hamster ovary (CHO) cells became thermotolerant after treatment with either heat for 10 min at 45.5 degrees C or incubation in 100 microM sodium arsenite for 1 h at 37 degrees C. Thermotolerance was tested using heat treatment at 45 degrees C or 43 degrees C administered 6-12 h after the inducing agent. At 45 degrees C thermotolerance ratios at 10(-2) isosurvival levels were 4.2 and 3.8 for heat and sodium arsenite, respectively. Recovery from heat damage as measured by resumption of protein synthesis was more rapid in heat-induced thermotolerant cells than in either sodium arsenite-induced thermotolerant cells or nonthermotolerant cells. Differences in inhibition of protein synthesis between heat-induced thermotolerant cells and sodium arsenite-induced thermotolerant cells were also evident after test heating at 43 degrees C for 5 h. At this temperature heat-induced thermotolerant cells were protected immediately from inhibition of protein synthesis, whereas sodium arsenite-induced thermotolerant cells, while initially suppressed, gradually recovered within 24 h. Furthermore, adding cycloheximide during the thermotolerance development period greatly inhibited sodium arsenite-induced thermotolerance (SF less than 10(-6] but not heat-induced thermotolerance (SF = 1.7 X 10(-1] when tested with 43 degrees C for 5 h. Our results suggest that both the development of thermotolerance and the thermotolerant state for the two agents, while similar in terms of survival, differed significantly for several parameters associated with protein synthesis. 相似文献
5.
Thermotolerance, the ability of cells and organisms to withstand severe elevated temperatures after brief exposure to mild elevated temperatures, has been studied in numerous laboratories. Survival thermotolerance is defined as the increase in cell or organism survival at severe elevated temperatures after a pretreatment at mild elevated temperatures. This study examines splicing thermotolerance in Drosophila melanogaster, the ability to splice pre-mRNAs made at the severe temperature (38 degrees C) after a brief pretreatment at a milder temperature (35 degrees C). It is probably one of a number of mechanisms by which cells adapt to heat shock. These experiments demonstrate that pre-mRNAs synthesized at the severe temperatures in splicing thermotolerant cells, although protected in splicing-competent complexes, are not actually processed to mature mRNAs until the cells are returned to their normal temperature. We have also studied the kinetics of acquisition and loss of splicing thermotolerance. As little as 10 min of pretreatment at 35 degrees C was sufficient to provide full splicing thermotolerance to a 30-min severe heat shock of 38 degrees C. Pretreatments of less than 10 min provide partial splicing thermotolerance for a 30-min severe heat shock. Full splicing thermotolerance activity begins to decay about 4 h after the cessation of the 35 degrees C incubation and is completely lost by 8 h after the pretreatment. The kinetics experiments of pre-mRNAs synthesized during the 38 degrees C treatment in splicing thermotolerant cells indicate that one or more splicing thermotolerance factors are synthesized during the 35 degrees C pretreatment which interact with pre-mRNA-containing complexes to keep them in a splicing-competent state. These kinetic experiments also indicate that in cells which are partially splicing thermotolerant, the pre-mRNAs synthesized early during the 38 degrees C incubation are protected, whereas those synthesized late are not. In the absence of splicing thermotolerant factors, the pre-mRNA-containing complexes leave the normal splicing pathway and are allowed to exit to the cytoplasm. 相似文献
6.
7.
Malik MK Slovin JP Hwang CH Zimmerman JL 《The Plant journal : for cell and molecular biology》1999,20(1):89-99
We have determined that one small heat shock protein gene, encoding Hsp17.7, plays an important role in the ability of carrot cells and plants to survive thermal stress. Transgenic cells and regenerated plants were generated in which the carrot Hsp17.7 gene was either constitutively expressed (denoted CaS lines) or expressed as a heat inducible antisense RNA (denoted AH lines). Thermotolerance measurements demonstrated that CaS lines were more thermotolerant than vector controls and AH antisense lines were less thermo- tolerant than vector controls. RNA analysis demonstrated that Hsp17. 7 mRNA was detectable, but not abundant, prior to heat shock in CaS cells, but not in vector control cells. Conversely, RNA analysis of antisense cells showed that, after heat shock, the amounts of mRNA for Hsp17.7 was moderately less abundant in AH cells than in vector controls. Analysis of protein synthesis in CaS cells did not indicate substantial synthesis or accumulation of Hsp17.7, or any small Hsp, at 23 degrees C. However, in the most thermotolerant line, protein synthesis was maintained at a higher rate than in other cell lines at a more extreme heat shock (42 degrees C). In contrast, antisense AH cells showed reduced synthesis of many Hsp, large and small. These results suggest that the Hsp17.7 gene plays a critical, although as yet not understood, role in thermotolerance in carrot. This represents the first demonstration of the ability to both increase and decrease thermotolerance by the manipulation of expression of a single gene. 相似文献
8.
Plants, as sessile organisms, employ multiple mechanisms to adapt to the seasonal and daily temperature fluctuations associated with their habitats. Here, we provide genetic and physiological evidence that the FtsH11 protease of Arabidopsis contributes to the overall tolerance of the plant to elevated temperatures. To identify the various mechanisms of thermotolerance in plants, we isolated a series of Arabidopsis thaliana thermo-sensitive mutants (atts) that fail to acquire thermotolerance after pre-conditioning at 38 degrees C. Two allelic mutants, atts244 and atts405, were found to be both highly susceptible to moderately elevated temperatures and defective in acquired thermotolerance. The growth and development of the mutant plants at all stages examined were arrested after exposure to temperatures above 30 degrees C, which are permissive conditions for wild-type plants. The affected gene in atts244 was identified through map-based cloning and encodes a chloroplast targeted FtsH protease, FtsH11. The Arabidopsis genome contains 12 predicted FtsH protease genes, with all previously characterized FtsH genes playing roles in the alleviation of light stress through the degradation of unassembled thylakoid membrane proteins and photodamaged photosystem II D1 protein. Photosynthetic capability, as measured by chlorophyll content (chl a/b ratios) and PSII quantum yield, is greatly reduced in the leaves of FtsH11 mutants when exposed to the moderately high temperature of 30 degrees C. Under high light conditions, however, FtsH11 mutants and wild-type plants showed no significant difference in photosynthesis capacity. Our results support a direct role for the A. thaliana FtsH11-encoded protease in thermotolerance, a function previously reported for bacterial and yeast FtsH proteases but not for those from plants. 相似文献
9.
In previous studies, we have demonstrated the differences in thermotolerance induced by heat and sodium arsenite (Lee et al., Radiat. Res. 121, 295-303, 1990). In this study, we investigated whether a 26-kDa protein might play an important role in evincing these differences. Chinese hamster ovary (CHO) cells treated for either 1 h with 100 microM sodium arsenite (ARS) or 10 min at 45.5 degrees C became thermotolerant to a test heat treatment at 43 degrees C administered 6 or 12 h later, respectively. After the test heating at 43 degrees C for 1.5 h, the level of 26-kDa protein in the nucleus was decreased by 92% in nonthermotolerant cells, 78% in ARS-induced thermotolerant cells, and 3% in heat-induced thermotolerant cells. Inhibiting protein synthesis with cycloheximide (CHM, 10 micrograms/ml) after ARS treatment eliminated thermotolerance to 43 degrees C and delayed restoration of the 26-kDa protein in the nucleus. In contrast, CHM neither prevented the development of thermotolerance nor inhibited the restoration of the 26-kDa protein in heat-induced thermotolerant cells. However, when cells were exposed to cold (4 degrees C), immediately after initial heating, restoration of the 26-kDa protein and development of thermotolerance did not occur. These results demonstrate a good correlation between the restoration and/or the presence of this 26-kDa protein and the development of protein synthesis-independent thermotolerance. 相似文献
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11.
Numerous studies have illustrated the need for antioxidant enzymes in acquired photosynthetic thermotolerance, but information on their possible role in promoting innate thermotolerance is lacking. We investigated the hypothesis that genotypic differences in source leaf photosynthetic thermostability would be dependent upon prestress capacity for antioxidant protection of the photosynthetic apparatus in Gossypium hirsutum. To test this hypothesis, thermosensitive (cv. ST4554) and reportedly thermotolerant (cv. VH260) G. hirsutum plants were exposed to control (30/20°C) or high‐day temperature (38/20°C) conditions during flowering and source leaf gas exchange, chlorophyll content and maximum photochemical efficiency (Fv/Fm) were measured for each treatment. The relationship between source leaf thermostability and prestress antioxidant capacity was quantified by monitoring the actual quantum yield response of photosystem II (PSII) (ΦPSII) to a range of temperatures for both cultivars grown under the control temperature regime and measuring antioxidant enzyme activity for those same leaves. VH260 was more thermotolerant than ST4554 as evidenced by photosynthesis and Fv/Fm being significantly lower under high temperature for ST4554 but not VH260. Under identical growth conditions, VH260 had significantly higher optimal and threshold temperatures for ΦPSII and glutathione reductase (GR; EC 1.8.1.7) activity than ST4554, and innate threshold temperature was dependent upon endogenous GR and superoxide dismutase (SOD; EC 1.15.1.1) activity. We conclude that maintaining a sufficient antioxidant enzyme pool prior to heat stress is an innate mechanism for coping with rapid leaf temperature increases that commonly occur under field conditions. 相似文献
12.
Heat can be one of the major abiotic stresses that adversely affect crop production worldwide at different stages of development.
As field screening for heat tolerance can be inconsistent and seasonally-limited, it is important to develop a reliable protocol
under controlled conditions that allows simultaneous screening of multiple genotypes. The objective of this research was to
develop a straightforward laboratory protocol using acquired thermotolerance (ATT) in peanut seedlings as a measure of one
mechanism of heat stress tolerance. Sixteen genotypes, including selected accessions of the US peanut minicore collection
along with standard checks, were evaluated for acquired themotolerance in two independent experiments. A change in the temperature
sensitivity of chlorophyll accumulation was used as an indicator of acquired thermotolerance. Pre-incubation at 38°C for 4 h
before the 30-min 50°C challenge triggered the acquired thermotolerance system of the leaf disks, resulting in chlorophyll
accumulation upon exposure to light. There was considerable variation among genotypes for ATT in both experiments. Genotypic
ranking for mean ATT values were highly correlated (0.949) in both experiments. The effect of seed weight on ATT was not significant.
This method is relatively simple and inexpensive and can be used to screen a large number of genotypes. 相似文献
13.
Genetics of thermotolerance in ciprofloxacin resistant mutant of Rhizobium leguminosarum bv phaseoli
Singh RK 《Indian journal of experimental biology》2001,39(8):818-820
A ciprofloxacin resistant mutant (Cf(R)) of Rhizobium leguminossarum bv phaseoli USDA 2695 which nodulates common bean plants (Phaseolus vulgaris L) was isolated after nitrous acid mutagenesis. Another mutant resistant to nalidixic acid (Nal(R)) was isolated spontaneously. Both mutants showed thermotolerance as evident by their ability to grow at elevated (40 degrees C) temperature, although the wild type (USDA 2695) failed to grow at this temperature. Transformation and plasmid curing experiments suggested the gene(s) controlling thermotolerance (TrR) and resistance to nalidixic acid or ciprofloxacin were located on the main chromosome and not on the plasmids. High frequency of co-transfer of TrR-Cr(R) and Tr(R)-Nal(R) during transformation experiments indicated a close association of these gene(s). Role of DNA gyrase and supercoiling in these thermotolerant mutants has been discussed. 相似文献
14.
Exposure of cells to heat induces thermotolerance, a transient resistance to subsequent heat challenges. It has been shown that thermotolerance is correlated in time with the enhanced synthesis of heat shock proteins. In this study, the association of induced heat shock proteins with various cellular fractions was investigated and the heat-induced changes in skeletal protein composition in thermotolerant and control cells was compared. All three major heat shock proteins induced in Chinese hamster fibroblasts after a 46 degrees C, 4-min heat treatment (70, 87, and 110 kDa) were purified with the cytoplasmic fraction, whereas only the 70-kDa protein was also found in other cell fractions, including that containing the cellular skeleton. Immediately after a second heat treatment at 45 degrees C for 45 min, the 110-kDa protein from thermotolerant cells also purified extensively with the cellular skeletal fraction. In this regard, the 110-kDa protein behaved similarly to many other cellular proteins, since we observed an overall temperature-dependent increase in the total labeled protein content of the high-salt-resistant cellular skeletal fraction after heat shock. Pulse-chase studies demonstrated that this increased protein content gradually returned to normal levels after a 3-hr incubation at 37 degrees C. The alteration or recovery kinetics of the total labeled protein content of the cellular skeletal fraction after heat shock did not correlate with the dramatic increase in survival observed in thermotolerant cells. The relationship between heat shock proteins and thermotolerance, therefore, does not correlate directly with changes in the heat-induced cellular alterations leading to differences in protein fractionation. 相似文献
15.
J Haveman A A Hart J Wondergem 《International journal of radiation biology and related studies in physics, chemistry, and medicine》1987,51(1):71-80
Cultured murine mammary carcinoma cells M8013 could be made thermotolerant by a priming heat treatment, 30 min at 43 degrees C, applied 5 h prior to subsequent heat treatment. The sensitivity of non-tolerant and thermotolerant cells to either radiation or heat combined with radiation was investigated. Analysis of survival curves with respect to D0 and N showed that thermotolerance had no influence on the radiation sensitivity of the cells. Thermal enhancement of radiation effects (in combined heat/irradiation treatments) was however reduced as a result of thermotolerance. When thermal enhancement ratios were (D0) plotted as a function of the cell killing effects of heat treatment alone, thermotolerance did not seem to have any influence. This latter observation suggests that thermotolerance modifies the effectiveness of the heat treatment for heat-induced cell lethality and radiosensitization equally. Comparison of our in vitro results with several in vivo data on normal tissues suggest that the reduction in 'effective' treatment temperature which has been observed in the in vivo studies as a result of thermotolerance may be explained by equal modification of the effects of heat by thermotolerance both for its direct effects and the radiosensitization. 相似文献
16.
Eukaryotic DNA polymerases delta and epsilon, both of which are required for chromosomal DNA replication, contain proofreading 3'-->5'exonuclease activity. DNA polymerases lacking proofreading activity act as strong mutators. Here we report isolation of thermotolerant mutants by using a proofreading-deficient DNA polymerase delta variant encoded by pol3-01 in the yeast Saccharomyces cerevisiae. The parental pol3-01 strain grew only poorly at temperatures higher than 38 degrees C. By stepwise elevation of the incubation temperature, thermotolerant mutants that could proliferate at 40 degrees C were successfully obtained; however, no such mutants were isolated with the isogenic POL3 strain. The recessive hot1-1 mutation was defined by genetic analysis of a weak thermotolerant mutant. Strong thermotolerance to 40 degrees C was attained by multiple mutations, at least one of which was recessive. These results indicate that a proofreading-deficient DNA delta polymerase variant is an effective mutator for obtaining yeast mutants that have gained useful characteristics, such as the ability to proliferate in harsh environments. 相似文献
17.
Heat-shock responses in two leguminous plants: a comparative study 总被引:16,自引:0,他引:16
Relative growth rates, basal and acclimated thermotolerance, membrane damage, fluorescence emission, and relative levels of free and conjugated ubiquitin and HSP70 were compared after 2 h of treatment at different temperatures between Prosopis chilensis and Glycine max (soybean), cv. McCall, to evaluate if the thermotolerance of these two plants was related to levels of accumulation of heat shock proteins. Seedlings of P. chilensis germinated at 25 degrees C and at 35 degrees C and grown at temperatures above germination temperature showed higher relative growth than soybean seedlings treated under the same conditions. The lethal temperature of both species was 50 degrees C after germination at 25 degrees C. However, they were able to grow at 50 degrees C after germination at 35 degrees C. Membrane damage determinations in leaves showed that P. chilensis has an LT(50) 6 degrees C higher than that of soybean. There were no differences in the quantum yield of photosynthesis (F(v)/F(m)), between both plants when the temperatures were raised. P. chilensis showed higher relative levels of free ubiquitin, conjugated ubiquitin and HSP70 than soybean seedlings when the temperatures were raised. Time-course studies of accumulation of these proteins performed at 40 degrees C showed that the relative accumulation rates of ubiquitin, conjugated ubiquitin and HSP70 were higher in P. chilensis than in soybean. In both plants, free ubiquitin decreased during the first 5 min and increased after 30 min of heat shock, conjugated ubiquitin increased after 30 min and HSP70 began to increase dramatically after 20 min of heat shock. From these data it is concluded that P. chilensis is more tolerant to acute heat stress than soybean. 相似文献
18.
Effect of pH on quercetin-induced suppression of heat shock gene expression and thermotolerance development in HT-29 cells. 总被引:3,自引:0,他引:3
Y J Lee L Curetty Z Z Hou S H Kim J H Kim P M Corry 《Biochemical and biophysical research communications》1992,186(2):1121-1128
When cells were heated for 15 min at 45 degrees C, they became thermotolerant to a second heat exposure at 45 degrees C. Thermotolerance developed rapidly, reached its maximum 6 hr after heat shock, and then gradually decayed. The development of thermotolerance was partially suppressed by treatment with various concentrations of quercetin (0.05-0.2 mM) at pH 7.4 after the initial heat treatment. In contrast, the drug markedly inhibited thermotolerance development at pH 6.5. Furthermore, a combination of low pH and quercetin treatment distinctively altered the expression of HSP70 gene compared with that of HSP28 or HSP90 gene. These results demonstrate a good correlation between the amount of HSP70 gene expression and development of thermotolerance. 相似文献
19.
Plants are constantly challenged with various abiotic stresses in their natural environment. Elevated temperatures have a detrimental impact on overall plant growth and productivity. Many plants increase their tolerance to high temperatures through an adaptation response known as acquired thermotolerance. To identify the various mechanisms that plants have evolved to cope with high temperature stress, we have isolated a series of Arabidopsis mutants that are defective in the acquisition of thermotolerance after an exposure to 38 degrees C, a treatment that induces acquired thermotolerance in wild-type plants. One of these mutants, atts02, was not only defective in acquiring thermotolerance after the treatment, but also displayed a reduced level of basal thermotolerance in a 30 degrees C growth assay. The affected gene in atts02 was identified by positional cloning and encodes digalactosyldiacylglycerol synthase 1 (DGD1) (the atts02 mutant was, at that point, renamed dgd1-2). An additional dgd1 allele, dgd1-3, was identified in two other mutant lines displaying altered acquired thermotolerance, atts100 and atts104. Expression patterns of several heat shock proteins (HSPs) in heat-treated dgd1-2 homozygous plants were similar to those from identically treated wild-type plants, suggesting that the thermosensitivity in the dgd1-2 mutant was not caused by a defect in HSP induction. Lipid analysis of wild-type and mutant plants indicated a close correlation between the ability to acquire thermotolerance and the increases in digalactosyldiacylglycerol (DGDG) level and in the ratio of DGDG to monogalactosyldiacylglycerol (MGDG). Thermosensitivity in dgd1-2 and dgd1-3 was associated with (1) a decreased DGDG level and (2) an inability to increase the ratio of DGDG to MGDG upon exposure to a 38 degrees C sublethal temperature treatment. Our results suggest that the DGDG level and/or the ratio of DGDG to MGDG may play an important role in basal as well as acquired thermotolerance in Arabidopsis. 相似文献
20.
Apoptosis constitutes a response of organisms to various physiological or pathological stimuli, and to different stresses. The ability of thermotolerance induced at a mild temperature of 40 degrees C to protect against activation of the apoptotic cascade by heat shock was investigated. When Chinese hamster ovary and human adenocarcinoma cervical cells were pretreated at 40 degrees C for 3 h, they were resistant to subsequent lethal heat shock at 43 degrees C. Induction of thermotolerance at 40 degrees C led to increased expression of heat shock proteins 27, 32, 72, and 90. Heat shock induced apoptotic events at the mitochondrial level, involving a decrease in membrane potential, translocation of Bax to mitochondria, and liberation of cytochrome c into the cytosol. These events were diminished in thermotolerant cells. Heat shock (42-45 degrees C) caused activation of initiator caspase-9 and effector caspases-3, -6, and -7, relative to controls at 37 degrees C. Activation of caspases was decreased in thermotolerant cells. Heat shock caused fragmentation of the caspase substrate, inhibitor of caspase-activated DNase. Fragmentation was diminished in thermotolerant cells. Thermotolerance afforded protection against heat shock-induced nuclear chromatin condensation, but not against necrosis. 相似文献