Comparative study on antioxidative system in normal and vitrified shoots of Populus suaveolens in tissue culture

To explore the physiological and biochemical mechanism of the occurrence of vitrified shoots of Populus suaveolens in tissue culture, the changes in water, chlorphyll, lignin, H₂O₂, phenylalanine ammonialyase (PAL), malonaldehyde (MDA), protective enzymatic systems, and some key enzymes involved in the ascorbate- glutathione cycle were comparatively studied in both normal and vitrified shoots of P. suaveolens. The results show that the lower activities of peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), glutathione reductase (GR) and PAL, and the less contents of chlorphyll, lignin, ascorbate (ASA) and reduced glutathione (GSH) as well as the lower ratios of ASA / DHA and GSH / GSSG are observed in vitrified shoots than in normal ones during the whole culture period. While in comparison with normal shoots, the higher activity of superoxide dismutase (SOD) and the more concentrations of water, H₂O₂, MDA, dehydroascorbate (DHA) and oxidized glutathione (GSSG) are found in vitrified shoots. Statistical analysis indicates that the enhanced activity of SOD and the decreased activities of CAT and POD as well as some enzymes involved in the ascorbate-glutathione cycle might be closely correlated to the accumulation of H₂O₂. The less regeneration of ASA and GSH and the lower capacity of the ascorbate-glutathione cycle observed in vitrified shoots might be due to a significant decrease in APX, MDAR, DHAR and GR activities and a decline in redox status of ASA and GSH. The decreases in chlorphyll content might result in a decline in photosynthesis. The lower activities of POD and PAL could result in the decrease of lignin synthesis and cell wall ligination, which might be the key factor leading to the increase in water content. It is concluded that the deficiency of detoxification capacity caused by the lower capacity of the ascorbate-glutathione pathway and the decreased activity of protective enzymatic system might lead to the large accumulation of H₂O₂ and the enhancement of membrane lipid peroxidation, which might be the main cause leading to the occurrence of vitrifying shoots of P. suaveolens in tissue culture. [Supported by National Natural Science Foundation of China (Grant No. 30271093) and the Foundation of State-designated Base for Biology Researching and Teaching in Beijing Forestry University]     

Comparative Study on Antioxidative System in Normal and Vitrified Shoots of Populus suaveolens in Tissue Culture

To explore the physiological and biochemical mechanism of the occurrence of vitrified shoots of Populus suaveolens in tissue culture, the changes in water, chlorphyll, lignin, H202, phenylalanine ammonialyase (PAL), malonaldehyde (MDA), protective enzymatic systems, and some key enzymes involved in the ascorbate- glutathione cycle were comparatively studied in both normal and vitrified shoots of P suaveolens. The results show that the lower activities of peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), glutathione reductase (GR) and PAL, and the less contents of chlorphyll, lignin, ascorbate (ASA) and reduced glutathione (GSH) as well as the lower ratios of ASA / DHA and GSH / GSSG are observed in vitrified shoots than in normal ones during the whole culture period, While in comparison with normal shoots, the higher activity of superoxide dismutase (SOD) and the more concentrations of water, H2O2, MDA, dehydroascorbate (DHA) and oxidized glutathione (GSSG) are found in vitrified shoots. Statistical analysis indicates that the enhanced activity of SOD and the decreased activities of CAT and POD as well as some enzymes involved in the ascorbate-glutathione cycle might be closely correlated to the accumulation of H202. The less regeneration of ASA and GSH and the lower capacity of the ascorbate-glutathione cycle observed in vitrified shoots might be due to a significant decrease in APX, MDAR, DHAR and GR activities and a decline in redox status of ASA and GSH. The decreases in chlorphyll content might result in a decline in photosynthesis. The lower activities of POD and PAL could result in the decrease of lignin synthesis and cell wall ligination, which might be the key factor leading to the increase in water content. It is concluded that the deficiency of detoxification capacity caused by the lower capacity of the ascorbate-glutathione pathway and the decreased activity of protective enzymatic system might lead to the large accumulation of H2O2 and the enhancement of membrane lipid peroxidation, which might be the main cause leading to the occurrence of vitrifying shoots of P.suaveolens in tissue culture.     

钙-钙调素对中东杨扦插苗的诱导抗冻作用

以零下低温锻炼和结合效应剂（CaCl2、钙离子螯合剂EGTA、钙离子通道阻断剂LaCl3或钙调素拮抗剂CPE）处理的低温锻炼下的中东扬扦插苗为试材，对其体内丙二醛（MDA）及钙调素CaM含量，SOD、POD及线粒体Ca^2-＋ATPase活性分别进行测定。结果表明，低温锻炼不仅在一定程度下提高了幼苗CaM含量，提高了SOD、POD和Ca^2＋-ATPase活性，降低了MDA含量；而且减少了低温胁迫所引起的SOD、POD，Ca^2＋＋ATPase和CaM含量的下降程度以及MDA的增加幅度。在低温锻炼的同时。用CaCl2处理能加强低温锻炼的效果，但这种效应可被EGTA、LaCl3或CPZ处理抑制。经低温锻炼后，幼苗中的CaM含量的增加有助于SOD、POD和线粒体Ca^2＋-ATPase活性的提高，进而对幼苗抗冻性的提高有明显的促进作用。     

Characterization and role of glucose-6-phosphate dehydrogenase of Populus suaveolens in induction of freezing resistance

Glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49) was purified from the leaves of 8-week-old Populus suaveolens cuttings. The enzyme activity in the absence and presence of reduced dithiothreitol (DTT_red) was determined. The results show that the G6PDH activity is not inactivated by pre-incubation with DTT_red, indicating that the purified enzyme probably presented in cytosol of P. suaveolens. The catalytic characteristics and kinetic parameters of cytosolic G6PDH purified from P. suaveolens cuttings were also studied. The results show that G6PDH is characterized by K _m value of 360 μmol·L⁻¹ for G6P and 16 μmol·L⁻¹ for NADP, a pH range of 7.3-8.9, and the maximum activity around pH 8.2. The enzyme activity is inhibited by various metabolites such as NADPH, NADH, GTP, UTP, ATP, AMP, ADP, CoA, acetyl CoA, fructose-6-phosphate (F6P), erythrose-4-phosphate (E4P), ribose-5-phosphate (R5P) and 3-phosphoglycerate (3-PG) (all at 1 mmol·L⁻¹ except for NADPH and NADH) to different extents. NADPH is the most effective inhibitor of enzyme activity, with an inhibition of 72.0%. The addition of metal ions such as MgCl₂, CaCl₂ and KCl (all 1.0 mmol·L⁻¹) to the standard reaction mixture has no remarkable influence on the cytosolic G6PDH activity. However, CdCl₂ (1.0 mmol·L⁻¹) causes high inhibitory effect on the enzyme activity. To explore the role of G6PDH on the enhancement of freezing resistance induced by freezing acclimation, the changes in the cytosolic G6PDH activity and freezing resistance (expressed as LT₅₀) of P. suaveolens cuttings during freezing acclimation at −20 °C were investigated. The results reveal that freezing acclimation decreases LT₅₀ of cuttings, and increases the activity of cytosolic G6PDH compared with control ones, while 2 d of de-acclimation at 25 °C result in a decrease in cytosolic G6PDH activity, and caused an increase in LT₅₀. Furthermore, the change in cytosolic G6PDH activity is found to be closely correlated to the degree of freezing resistance of cuttings during freezing acclimation. It is suggested that cytosolic G6PDH may be involved in the induction of freezing resistance of cuttings. [Supported by the Foundation of State-designated Base for Biology Researching and Teaching in Beijing Forestry University]     

CO_2浓度升高对落叶松叶片抗氧化系统的影响

在CO_2浓度升高和对照处理的两个开顶式气室(OTC-Ⅰ型)内,研究分析了高浓度CO_2对落叶松针叶抗氧化系统酶活力的影响。结果表明:短期内(2个月左右)CO_2浓度升高条件下,超氧化物歧化酶(SOD)、抗坏血酸(ASA)、抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)含量上升,且SOD与O_2·~-产生速率之间呈极显著负相关;但长期高浓度CO_2处理下,ASA含量与SOD、APX、GR活性略有下降,可见,较长期高浓度CO_2处理可能使此效果发生逆转。在CO_2浓度升高条件下,植物可能通过抗氧化能力的增强来减轻胁迫造成的细胞损伤,从而增强植物的抗性。     

Responses of antioxidant enzymes in the needles of hinoki cypress (Chamaecyparis obtusa) seedlings to nutrient solutions containing various calcium/aluminum ratios

An attempt was made to examine the possible connection between the various ratios of calcium/aluminum (Ca/Al) in the nutrient solution of plant cultures and the active oxygen scavenging system of hinoki cypress (Chamaecyparis obtusa) seedlings. The hinoki cypress seedlings were transferred to nutrient solutions containing 5 mM AlCl₃ together with various concentrations of Ca(NO₃)₂ in pots containing glass beads and Teflon tips. The activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR) in the needles were estimated at several stages during the 7-day treatment. The samples treated with the lower Ca/Al solutions show the highest SOD activities. The activities of APX and GR, both of them involved in the ascorbate-glutathione cycle, showed the same tendency with decrease to higher Ca/Al ratio. These results indicate that rhizospheric Ca might compete with Al and ameliorate Al toxicity on and in the roots, the Al stress is not transformed to the needles after a few days, and the ascorbate-glutathione cycle in the hinoki cypress needles might fluctuate and be suppressed by the rhizospheric Al stress during the 7 days. This work was supported in part by funding from the Japan Science & Technology Corporation, the CREST program 1996–2001, and the Center for Forest Decline Studies.     

渗透胁迫和钙处理对杉木幼苗膜脂过氧化及保护酶活性的影响

杉木幼苗用ＰＥＧ６０００渗透胁迫（－１．５ＭＰａ,－２．５ＭＰａ）,在４８ｈ内其叶片中Ｈ２Ｏ２含量和ＭＤＡ含量明显增加,Ｈ２Ｏ２含量从３５．５μｍｏｌ．ｇ＾－１分别增加到４３．０μｍｏｌ．ｇ＾－１和４８．０μｍｏｌ．ｇ＾－１;ＭＤＡ含量从１７０．５μｍｏｌ．ｍｇ＾－１分别增加到３４９．５μｍｏｌ．ｍｇ＾－１和３６５．２μｍｏｌ．ｍｇ＾－１,其变化与胁迫强度成正比;４８ｈ后Ｈ２Ｏ２含量下降,但ＭＤＡ     

硒对粗壮女贞新梢和生理参数的影响

采用国际通用营养液配方,以砂培方式研究不同质量浓度硒对粗壮女贞叶绿素、根系活力、SOD、POD、CAT、GR、MDA以及新梢萌芽能力的影响,结果表明：硒质量浓度为1mg／L时,SOD、POD活性最弱,根系活力最强,可促进粗壮女贞的生长,增强新梢的萌芽能力。而当硒质量浓度大于1mg／L时,则会起到抑制作用;MDA随着硒质量浓度的增大呈升高趋势,CAT和GR活性随着硒质量浓度的增加而减弱。因此认为适量的硒处理能够增强粗壮女贞的抗氧化能力,提高产量。     

Rapid predictions of cold tolerance in Douglas-fir seedlings using chlorophyll fluorescence after freezing

Chlorophyll fluorescence measurements were performed on the foliage of 3-year-old (11/2+11/2) nursery-grown Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco] seedlings after exposure to controlled freezing temperatures, in the laboratory, to assess low temperature tolerance. The seedlings were propagated in an Irish nursery and lifted at monthly intervals overwinter 1999 and 1999–2000. Excised shoots from first-order laterals were frozen, in the dark. After freezing, needles were immediately assessed using chlorophyll fluorescence. The excised shoots were then maintained under controlled conditions for 14 days and visually assessed for needle damage. The chlorophyll fluorescence parameter, F _v/F _m, accurately predicted cold hardiness and was linearly related to visual needle damage and short-term survival. An equation was constructed using F _v/F _m data for determining the LT₅₀, that is, the freeze temperature causing 50% seedling damage. The predictions of F.LT₅₀ (fluorescence-based empirical determination of LT₅₀) have been tested over two seasons (i.e., against a second independent data set) with variability between 0 and 1.8°C of visual estimates, though predictions were often 1.1°C of the visual assessment. This approach provided a simple, rapid and accurate prediction of cold tolerance, under climatic conditions where in situ measurements are unreliable. The method can be used to predict if Douglas-fir seedlings have developed sufficient tolerance for lifting to the cold-store, or for planting.     

三年桐、千年桐感染枯萎病病原菌后的生理反应

枯萎病是油桐毁灭性病害。中国广泛种植的油桐有三年桐和千年桐,三年桐易感枯萎病,千年桐抗枯萎病。为探讨三年桐与千年桐在枯萎病应答过程中的差异,在对油桐枯萎病灾区调研的基础上,进行枯萎病病原菌的分离、鉴定,并进一步探讨三年桐、千年桐接种枯萎病病原菌后超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性和丙二醛(MDA)含量的变化。结果表明:油桐枯萎病病原菌为尖孢镰刀菌,接种试验显示千年桐在接种尖孢镰刀菌后SOD、POD、CAT较三年桐均呈现较高活性,在接种后SOD、POD活性先升高后降低,CAT活性升高并维持高活性,MDA含量变化不明显;三年桐SOD活性和MDA含量均先升高后降低,POD活性先降低再升高,CAT活性先降低后升高又降低。千年桐作为抗病种,其抗病性可能与其本身具有较高的SOD、POD、CAT活性有关,并且和病原菌感染后酶活上升有关;三年桐为易感病种,接种后SOD、POD、CAT活性有变化,但仍不能抵御病原菌的危害。     

Effects of cold-hardening on compatible solutes and antioxidant enzyme activities related to freezing tolerance in Ammopiptanthus mongolicus seedlings

Cold acclimation is associated with many metabolic changes that lead to an increase of freezing tolerance. In order to investigate the biochemical process of cold acclimation in Ammopiptanthus mongolicus, seedlings were acclimated at 2℃ under 16-h photoperiod （150 μmol·m^-2·s^-1 photosynthetically active radiation） for 14 d. Freezing tolerance in seedlings increased after 14 d of cold-hardening. Contents of protein, proline and solute carbohydrate in cotyledon increased after cold acclimation. Patterns of isozymes of superoxide dismutase （SOD）, peroxidase, catalase and polyphenol oxidase （PPO） were investigated. The activities of SOD, peroxidase and PPO in cold acclimated plants were increased during cold-hardening. We deduced that compatible solutes and antioxidant enzymes play important roles in development of freezing tolerance during cold acclimation in this evergreen woody plant.     

Chlorophyll fluorescence and variations in tissue cold hardiness in response to freezing stress in Douglas-fir seedlings

Two-year-old coastal Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) seedlings from two seed lots were exposed to controlled freezing temperatures every 4 weeks from October 1993 through April 1994. Freezing effects were assessed by measuring chlorophyll fluorescence emissions 1 day after freezing and by evaluating damage to the bud, cambium, and needle tissues 7 days after freezing. Differences between the seed lots as well as changes in cold hardiness among the bud, cambium, and needle tissues were evident throughout the duration of the study. Tissue damage was higher with increased freezing stress. Severity of damage to each of the tissues varied seasonally. Chlorophyll fluorescence emissions were lower with higher freezing stress (except during November and December, when test temperatures were not low enough to significantly damage the seedlings) and showed a strong relationship with morphological assessments of freezing stress. The slope of the slow kinetics phase of the chlorophyll fluorescence curve tended to be less steep (i.e., quenching was reduced) with higher freezing stress. Nonfrozen chlorophyll fluorescence measurements showed no obvious relationship with LT₅₀ for either seed lot. However, chlorophyll fluorescence measurements are useful for determining cold hardiness and resistance to stress, because they provide a rapid assessment of seedling vigor following exposure to freezing.     

低温锻炼对冷胁迫下油茶幼苗光合速率与抗氧化酶活性的影响

以油茶品种"杂优闽1"为试验材料,以未经低温锻炼的油茶幼苗为对照,研究了油茶幼苗在(10±0.5)℃低温锻炼、(2±0.5)℃冷胁迫和常温恢复期间的光合速率和抗氧化酶活性的变化。结果表明:低温锻炼3 d可明显提高油茶幼苗的抗寒性;冷胁迫3 d,经低温锻炼后的油茶幼苗叶片净光合速率和相对含水量明显高于对照;且其在常温下的恢复也较对照迅速。在低温锻炼和冷胁迫期间,油茶幼苗叶片脯氨酸和可溶性糖含量明显增加,且丙二醛(MDA)含量明显低于对照;经低温锻炼的油茶幼苗叶片超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)活性则均明显高于未经低温锻炼的油茶幼苗,说明渗透调节物质含量增加和保护酶活性增强在低温锻炼诱导油茶幼苗的抗寒性上发挥着重要的作用。     

在低温锻炼中毛白杨幼苗可溶性总蛋白、RNA、DNA、RNase及抗冻性的变化

低温锻炼不仅提高了毛白杨幼苗存活率和抗冻性以及RNA和可溶性总蛋白的含量 ,降低了RNase活性 ,而且能减轻低温胁迫引起的RNA和可溶性总蛋白含量的下降程度和RNase活性的提高 ,有利于幼苗在恢复过程中RNA和可溶性总蛋白水平的迅速回升以及RNase活性的降低 .进一步研究发现 ,DNA含量无论在低温锻炼中还是在低温胁迫下或是在随后的恢复生长期均保持相对稳定 ;低温锻炼所引起的RNA含量的增加 ,与RNase活性的降低呈明显的负相关 ,与可溶性总蛋白含量的增加以及幼苗抗冻性的提高成正相关 .这表明低温锻炼可能抑制RNase活性 ,有效地促进RNA含量的增加 ,而RNA可能参与了可溶性总蛋白的合成及抗冻性的诱导     

Cold acclimation induced accumulation of phenolic compounds and freezing tolerance in Ammopiptanthus mongolicus

Ammopiptanthus mongolicus, the only freezing tolerant evergreen broad-leaved shrub, local species of the Alashan desert, northwest sand area of China, can survive -30°C or even lower temperature in winter. In the present study, the secondary products phenolics in A. mongolicus cotyledons were determined to study the effects of phenolics on cold tolerance. Cytochemical localization of phenolics in cotyledon cells was observed by electron microscopy and the content of phenolic compounds was assayed by spec-trophotometric measurement. The results showed that the freezing tolerance of A. mongolicus seedlings increased after acclimation at 2-6°C for 14 days, which accompanied the increase of the content of phenolic compounds in cotyledons. Cytochemical observation showed that phenolic deposits were mainly localized in vacuoles and in close proximity to tonoplast, and also in the cytoplasm. The amount and the size of phenolics droplets increased obviously in cytoplasm and vacuoles after cold acclimation, predominantly ag-gregated along membranes of vacuoles and tonoplast. No phenolic deposits were found in cell walls. As hydrogen-or elec-tron-donating agents, phenolics may protect plant cells against reactive oxygen species formed during chilling or freezing stress and improve the freezing tolerance of cold-acclimated A. mongolicus seedlings.     

Effects of mechanical damage and herbivore wounding on H2O2 metabolism and antioxidant enzyme activities in hybrid poplar leaves

The changes of hydrogen peroxide (H₂O₂) metabolism and antioxidant enzyme activities in a hybrid poplar (Populus simonii × P. pyramidalis ‘Opera 8277’) in response to mechanical damage (MD) and herbivore wounding (HW) were investigated to determine whether H₂O₂ could function as the secondary messenger in the signaling of systemic resistance. Results show that H₂O₂ was generated in wounded leaves through MD and HW treatments and systemically in unwounded leaves around the wounded leaves. The activities of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) were also enhanced. However, the H₂O₂ accumulation and antioxidant enzyme activities were inhibited in MD leaves through the pretreatment with DPI (which is a specific inhibitor of NADPH oxidase). The results of this study suggest that H₂O₂ could be systemically induced by MD and HW treatments, and H₂O₂ metabolism was closely related to the change in SOD, APX and CAT activities. A high level of antioxidant enzymes could decrease membrane lipid peroxidation levels and effectively induce plant defense responses. Foundation project: This research is supported by the Key Science Program of the Sate Forestry Administration of China (2006–59), and the National Key Project of Scientific and Technical Supporting Programs Funded by Ministry of Science & Technology of China (2006BAD01A15; 2006BAD24B04). Biography: AN Yu (1982–), female, Postgraduate in College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, P. R. China.     

水分胁迫对板栗幼苗叶片活性氧代谢的影响

采用盆栽控水试验测定了板栗幼苗叶片的生理生化指标,结果表明:在土壤水分胁迫下,随胁迫强度的增加,板栗叶片相对含水量和可溶性蛋白含量呈逐渐减小趋势;总叶绿素、叶绿素a、叶绿素b及类胡萝卜素含量呈下降趋势,但下降幅度不同;活性氧保护酶SOD、POD、CAT随胁迫程度的增强呈先降后增再降低的变化趋势,而ASP基本呈降低的变化趋势;O-·2产生速率呈先增再降后增加的变化趋势,而H2O2含量呈先增后降的变化趋势,其中中度胁迫时含量最大;MDA含量与质膜相对透性均随胁迫程度的增强而增大;SOD、CAT、ASP等保护酶可以作为板栗幼苗抗旱力强弱的评价指标.     

Comparison of G6PDH Activity and LT50 Between P.tomentosa and P.suaveolens During Freezing Acclimation

The differences of glucose-6P dehydrogenase(G6PDH) activity and freezing resistance induced by freezing acclima-tion between cuttings of freezing-sensitive P.tomentosa and freezing-resistant P.suaveolens were compared for exploring the role of G6PDH on the enhancement of freezing resistance induced by freezing acclimation.After 5d of freezing acclimation at -3℃ ,the LT50 of P.tomentosa has deereased from -6.2℃ incontrol cuttings to -14.3℃ in freezing acclimated ones,and the increase of LT50 of P.tomentosa has decreased from -6.2℃ in control cutting to -14.3℃ in freezing acclimated ones,and the increase of G6PDH activity was observed in freezing acclimated cuttings as compared with control ones.Whereas,when P.suaveolens was freezing acclimated at -20℃ for 5d,the LT50 has decreased from -27.1℃ in control cuttings to -43.5℃ in freezing acclimated ones,and the activity of G6PDH increased considerably.In addition,the increase of LT50 and the decrease of G6PDH activity resulting from 2d of deacclimation at 25℃ were found in two kinds of freezing acclimated cuttings.It is concluded that the increase in the activity of G6PDH may associate with the inherited freezing resistance of species and the enhancemen of freezing resistance of cuttings,and may play an important role in the antifreeze process under freezing temperature,which would provide the basis for the study on the molecular mechanism of freezing resistance in P.suaveolens and the cloning of gene associated with freezing resistance.     

毛白杨幼苗抗氧化系统对盐胁迫的响应

在 3种不同浓度的NaCl (85mmol·L-1,170mmol·L-1,2 6 0mmol·L-1)胁迫下 ,对毛白杨幼苗的超氧化物歧化酶 (SOD)和过氧化物酶 (POD)活性以及丙二醛 (MDA)含量进行测定 .结果发现 ,在NaCl胁迫 3小时后 ,幼苗SOD和POD活性显著降低 ,随着胁迫时间的延长 ,这两种酶活性降低较缓慢 ,但SOD活性下降程度均较POD小 .在NaCl胁迫中 ,MDA含量明显增加 ,并于胁迫 15小时后达到最大值 ,随后含量增加较小 .进一步分析发现 ,不同浓度NaCl处理对SOD和POD活性以及MDA含量的作用效应是不同的 ,其中 2 6 0mmol·L-1NaCl处理明显促进幼苗SOD和POD活性的下降与MDA含量的增加 ;而且MDA含量与SOD和POD活性的下降有关 .结果表明 ,NaCl胁迫所引起的SOD和POD活性下降可能加强膜脂过氧化作用 ,最终导致幼苗MDA含量的增加与伤害的发生     

First frost: Effects of single and repeated freezing events on acclimation in Picea abies and other boreal and temperate conifers

In experiments with needles of Picea abies, we tested the specific hypothesis that a single night of freezing acts as a signal that triggers a rapid increase in low temperature (LT) tolerance, and the more general hypothesis that repeated or prolonged freezing stimulates increased LT acclimation. In three growth chamber experiments involving acclimation under early- to mid-autumn light and temperature conditions followed by one or more freezing treatments, we found no significant effect of a single night of freezing on LT tolerance, and only limited and inconsistent effects of repeated and prolonged freezing. We also tested the effect of prolonged storage at −5 °C on LT tolerance on samples of three boreal and three temperate conifer species during acclimation under field conditions, and again found no consistent enhancement of LT tolerance attributable to freezing in either group. In agreement with our own and others’ anecdotal observations that some species can attain nearly maximal LT tolerance in the absence of freezing under field conditions, we conclude that freezing is neither required nor a major influence in LT acclimation, at least in well-studied boreal conifer species, while the effects of freezing on temperate conifers are not as well-documented. We conclude that freezing treatment of conifer seedlings to ensure sufficient hardiness for late planting seems to offer little practical advantage.