圆柏属常绿木本植物叶片水分、渗透调节物质的季节变化与抗冷冻性的关系

以祁连圆柏（Sabina przewalskii）和圆柏（Sabina chinensis）为材料,研究了圆柏属植物的抗冷冻性与叶片水分和渗透调节物质季节变化的关系。结果表明,随秋季气温下降,叶片组织相对含水量（RWC）和自由水含量（FWC）下降,束缚水含量（BWC）、脯氨酸（Pro）、游离氨基酸（FAA）、可溶性蛋白质（SP）和可溶性糖（SS）含量增加,翌年春季气温回升,相对含水量和自由水含量增加,束缚水含量下降,有机渗透调节物再次增加。有机渗透调节物在秋末冬初及初春的增加正好与圆柏属植物抗冻锻炼及脱冻恢复生长的时间相吻合,因而是植物抗冻及脱冻适应的重要生理响应,在降低细胞冰点、防止细胞结冰引起的膜机械伤害、抑制膜脂过氧化、保护膜稳定性方面具有重要作用。祁连圆柏的相对含水量和自由水含量均低于圆柏,3种有机渗透调节物含量均高于圆柏,表明祁连圆柏在抗冷冻性诱导中具有更广泛的适应性策略。     

圆柏属常绿木本植物元素含量的季节动态

测定分析了圆柏属2种常绿木本植物叶片干物质含量、叶绿素含量及N、P、K、Na、Ca、Mg等元素含量的季节变化和元素间的关系。结果发现,两种圆柏叶片N含量与叶绿素含量正相关,反映了其一年四季的生长情况;两树种N和K含量的季节变化趋势相似,夏、秋季高于冬、春季,表明两树种夏、秋季富集的营养物质较多,与夏、秋季较高的生物量一致;而P、Ca、Mg和Na含量是冬、春季较夏、秋季高,表明圆柏属植物通过积累这些无机渗透调节剂来增强其对低温的抗性。两树种叶片对N和K同步积累,对P、Na和Mg同步积累,但在元素积累模式上也存在一定的差异,元素间的关系也要复杂得多。     

高寒山区植物根抗氧化酶系统的季节变化与抗冷冻关系

在高寒山区（海拔2900m）和选取4种多年生草本植物,即无芒雀麦（Bromus inermis）、草地早熟禾（Poa sphyondylodes）、花誉麦（Bromus sinensis）和垂重申披碱草（Elymus nutans）,测定了秋末、冬初、冬季、春季气温变化过程中其根中丙二醛（MDA）含量和抗氧酶活力（过氧化氢酶（CAT）、过氧化物酶（POD）、超氧物歧化酶（SOD））和抗坏血酸氧化酶（APX）变化,分析了抗氧酶系统在根抗冷适应中的作用,结果表明,随秋末降温植物根中MDA含量增加,尔后下降,在冬季和翌年春季保持相对稳定。从9月初到10月下旬,4种植物根中SOD、CAT、POD活力平均增加170％、130％和56％。在冬季下降,但仍远高于9月,在春季气温上升过程中酶活力上升。根能在组织结冰状况下生存与其具备完善的保护酶系统,能及时清除氧自由基抑制膜脂过氧化维持膜完整性有关,据降温过程中MDA含量和抗氧酶活力变化,可将根冷适应分为两个阶段,即第1阶段平均气温在0℃以上,抗氧酶活力增强,MDA增加阶段,第2阶段平均气温降至0℃以下,最低气温降到－15℃以下,抗氧酶活力下降,MDA无明显变化阶段。     

高温胁迫对两种常绿杜鹃亚属植物幼苗生理生化特性的影响

为探明常绿杜鹃亚属植物高温致伤的生理机制,以猴头杜鹃（Rhododendron simiarum Hance）、井冈山杜鹃（R.jinggangshanicumTam）四年生实生苗为材料,通过人工气候箱中的盆栽实验,研究了不同程度的高温胁迫对两种常绿亚属杜鹃幼苗叶片生理生化特性的影响。结果表明：（1）两种杜鹃叶片的丙二醛（MDA）、过氧化氢（H2O2）及脯氨酸（Pro）、可溶性蛋白质含量均随胁迫温度的升高而增大;猴头杜鹃的H2O2、可溶性蛋白含量及增幅均明显小于井冈山杜鹃,而MDA、Pro的增幅则因胁迫温度而异。（2）两种杜鹃叶片抗坏血酸（AsA）含量均随胁迫温度的升高而降低,但猴头杜鹃的降幅小于井冈山杜鹃。（3）高温胁迫下,猴头杜鹃除过氧化氢酶（CAT）活性增幅略小外,超氧化物歧化酶（SOD）、过氧化物酶（POD）及抗坏血酸过氧化物酶（APX）增幅均大于井冈山杜鹃。可见,随着胁迫温度的升高两种杜鹃叶片膜脂过氧化作用加重,而猴头杜鹃较井冈山杜鹃具有更强的酶促和非酶促清除活性氧能力。     

外源水杨酸对低温胁迫下圆柏属植物幼苗生理特性的影响

以3年生圆柏和祁连圆柏幼苗为材料,采用不同浓度水杨酸(SA)预处理两种圆柏属植物幼苗,测定-4℃低温胁迫处理第6天叶片相对含水量(RWC)、相对电导率(REC)、丙二醛(MDA)、可溶性糖(SS)、可溶性蛋白(SP)、脯氨酸(Pro)和类胡萝卜素(Car)含量及超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、抗坏血酸氧化酶(APX)和谷胱甘肽还原酶(GR)活性,分析外源 SA 对低温胁迫下圆柏属植物叶片膜脂过氧化、渗透调节物质及抗氧化酶系统的影响,为培育较多品种的抗冷冻常绿植物提供理论依据。结果表明：低温胁迫下,一定浓度的 SA 预处理能有效保护幼苗叶片膜系统的稳定性、增加渗透调节物质含量和提高抗氧化酶活性,其中200 mg/L SA 预处理对提高圆柏抗寒性效果最好,30 mg/L SA 预处理对提高祁连圆柏抗寒性效果最好,且在外源 SA 预处理下,祁连圆柏的抗寒性比圆柏的强。因此,施用适合浓度的 SA 在提高圆柏属植物抗寒性方面具有较好的应用价值。     

不同海拔青海云杉与祁连圆柏叶片抗氧化系统

以祁连山寺大隆林区连续海拔梯度(2 665~3 365 m)上青海云杉(Picea crassifolia)和祁连圆柏(Sabina przewalskii)为材料, 测定叶片中抗氧化保护系统的变化, 探讨常绿木本植物抗氧化系统对高山极端环境的适应机制。结果显示, 祁连圆柏和青海云杉叶片中丙二醛(MDA)含量均与海拔高度呈正相关, 相同海拔上青海云杉MDA含量极显著高于祁连圆柏(p＜0.01)。随海拔升高, 两树种抗氧化保护酶超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)活性和非酶促抗氧化剂脯氨酸(Pro)、抗坏血酸(AsA)、还原性谷胱甘肽(GSH)含量均呈明显增加趋势。青海云杉叶片的AsA水平高于祁连圆柏, 但对海拔变化的敏感性较低; 祁连圆柏的GSH、Pro水平及其对海拔变化的敏感性均高于青海云杉。结果表明, 研究区青海云杉所受过氧化伤害较祁连圆柏更严重, 但两树种清除O₂^-·的能力相当而主要负责分解H₂O₂的酶种有所不同: 祁连圆柏中为POD, 青海云杉中则为CAT、APX和GR, AsA-GSH循环系统在青海云杉活性氧清除中的作用强于在祁连圆柏中, 祁连圆柏的活性氧清除物质可能以Pro为主。     

井冈山杜鹃叶片抗氧化系统对高温胁迫的响应

以井冈山杜鹃(Rhododendron jinggangshanicum Tam)4年生实生苗为材料,通过人工气候箱盆栽实验,研究了不同高温处理下其叶片形态及生理生化指标的变化,以阐明高温胁迫下井冈山杜鹃抗氧化系统的防御机理.结果显示:(1)30℃和38℃处理6 d后,叶片过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)及过氧化氢酶(CAT)活性分别比对照极显著(P<0.01)升高44.44%和127.16%、33.12%和129.35%、106.52%和188.64%;超氧化物歧化酶(SOD)活性增幅较小,分别比对照增加3.77%和5.11%.(2)随着胁迫温度的升高,可溶性蛋白质、脯氨酸(Pro)含量均明显上升,而抗坏血酸(AsA)含量却显著降低;除30℃处理与对照之间的可溶性蛋白质含量差异显著(P<0.05)外,其它处理间差异均达到极显著(P<0.01).(3)随着胁迫温度的升高,叶片过氧化氢(H2O2)、丙二醛(MDA)含量大量积累,各处理间差异达到极显著或显著;而形态上则表现为受害加重,植株出现轻度萎蔫、老叶褐化脱落、新叶坏死等现象.研究表明,井冈山杜鹃受到高温胁迫的氧化伤害可能与AsA含量显著下降和H2O2、MDA含量大幅增加有关,其植物体能诱导增加抗氧化保护酶活性及Pro含量以提高其抗氧化能力,从而减轻高温引发的活性氧毒害.     

低温胁迫对两种圆柏属植物亚细胞抗氧化酶活性的影响

以祁连圆柏和圆柏幼苗为材料,研究不同处理时间下低温胁迫对圆柏属植物叶片亚细胞抗氧化酶活性的影响,探讨其在圆柏属植物叶片中的亚细胞定位。结果表明：低温胁迫下,丙二醛(MDA)含量和抗氧化酶活性随时间变化均呈先升后降趋势,祁连圆柏中抗氧化酶的种类比圆柏的多且活性强,而 MDA 含量低于圆柏,表明祁连圆柏在低温胁迫下具有更广泛的适应性。此外,两种圆柏植物叶片超氧化物歧化酶(SOD)和抗坏血酸过氧化物酶(APX)定位为叶绿体＞细胞溶质＞线粒体,过氧化氢酶(CAT)定位为线粒体＞叶绿体＞细胞溶质,谷胱甘肽还原酶(GR)定位为线粒体＞细胞溶质＞叶绿体,祁连圆柏过氧化物酶(POD)定位为细胞溶质＞叶绿体＞线粒体,圆柏POD定位为细胞溶质＞线粒体＞叶绿体,且抗氧化酶SOD、APX和 GR在亚细胞中分布差异达到极显著,这说明抗氧化酶在其中一种亚细胞中发挥主要作用,为克隆亚细胞组分中的抗氧化酶基因提供了理论依据。     

大气CO2浓度升高对毛竹叶片膜脂过氧化和抗氧化系统的影响

为了解竹子对大气CO2浓度升高的生理响应,为气候变化背景下的竹林适应性管理提供理论依据,运用开顶式气室(OTCs)设置了3个CO2处理浓度(环境大气、500和700μmol·mol-1),探讨了大气CO2浓度升高对毛竹(Phyllostachys edulis)叶片光合色素、膜脂过氧化和抗氧化系统的影响。结果表明:与环境大气比较,500μmol·mol-1浓度处理30d时对毛竹叶片光合色素、膜脂过氧化和抗氧化系统影响并不明显,仅叶片CAT活性显著降低;随着处理时间的延长,对毛竹叶片膜脂过氧化和抗氧化系统的影响逐渐显现,至处理90d时,除叶片可溶性糖含量无明显变化,其他测定指标均有显著变化;700μmol·mol-1浓度处理在不同时间上对毛竹叶片膜脂过氧化和抗氧化系统的影响均较500μmol·mol-1CO2浓度处理明显,处理30d时毛竹叶片可溶性糖含量和抗氧化酶活性有明显变化,处理90d时,各项测定指标均有显著变化;研究表明大气CO2浓度升高一定程度上能增强毛竹的抗氧化能力,但光合产物的过量积累也会造成碳水化合物源-库失衡和Rubisco的再生受到反馈作用抑制。     

钼,硼对大豆叶片膜脂过氧化及体内保护系统的影响

以３个大豆（Ｇｌｙｃｉｎｅ ｍａｘ Ｌ．）品种（“浙春３号”、“浙春２号”和“３８１１”）为材料,设置不同的钼、硼水平,研究了３个生育期（五叶期、初花期和盛荚期）大豆叶片膜脂过氧化及体内保护系统的变化。结果表明：在低钼、低硼胁迫下,大豆叶片的质膜透性（ＭＰ）、丙二醛（ＭＤＡ）和脯氨酸（Ｐｔｏ）的含量、多酚氧化酶（ＰＰＯ）和抗坏血酸氧化酶（ＡＯ）的活笥增加,抗坏血酸（ＡｓＡ）的含量及超氧化物歧化酶（     

两种圆柏属植物不同季节显微和超微结构变化与耐寒性的关系

以祁连圆柏和圆柏为材料,研究了圆柏属植物的抗寒性与解剖结构变化,以及淀粉与可溶性糖粉转化的关系.结果表明,祁连圆柏和圆柏叶片的上下表皮细胞外覆盖一层角质膜,叶肉细胞间隙大,形成发达的通气组织.在生长季节,二者叶绿体内均积累了很多淀粉粒,在冷冻休眠季节淀粉粒消失或变小,同时叶片可溶性糖分含量增加,祁连圆柏的可溶性糖分增幅高于圆柏;而休眠季节圆柏叶绿体受到低温伤害,脂质球数目增多,出现脂质小滴,部分叶绿体变形,祁连圆柏类囊体的结构受到影响较小,脂质球数量少,叶绿体形状没有发生明显的变化.因此,祁连圆柏和圆柏抗寒性与发达的通气组织和淀粉粒生长季节的积累有关,在低温胁迫下祁连圆柏叶绿体表现出高的稳定性,通过可溶性糖分的调节力强,耐寒性增强.     

Environmental regulation and physiological basis of freezing tolerance in woody plants

Cold acclimation of plants is a complex process involving a number of biochemical and physiological changes. The ability to cold acclimate is under genetic control. The development of freezing tolerance in woody plants is generally triggered by non-freezing low temperatures but can also be induced by mild drought or exogenous abscisic acid, as well as by short photoperiod. In nature, the extreme freezing tolerance of woody plants is achieved during sequential stages of cold acclimation the first of which is initiated by short photoperiods and non-freezing low temperatures, and the second by freezing temperatures. Although recent breakthroughs have increased our knowledge on the physiological molecular basis of freezing tolerance in herbaceous species, which acclimate primarily in response to non-freezing low temperatures, very little is known about cold acclimation of woody plants. This article attempts to review our current understanding of the physiological aspects that underline cold acclimation in woody plants.     

Seasonal changes in the physical state of crown water associated with freezing tolerance in winter wheat

The relationship between freezing tolerance (expressed as LT₅₀, the lethal freezing temperature for 50% of plants) and the amount and physical state (as determined by spin-lattice [T₁] and spin-spin [T₂] relaxation times of protons) of water in crown tissue was examined in contrasting winter wheat (Triticum aestivum L.) varieties grown under field conditions from 1992 to 1994. During acclimation, the LT₅₀ values decreased from around -7 to -17, -20 and -27°C in PI 173438, Chihokukomugi and Valuevskaya, respectively. Tissue water content decreased continuously through autumn to reach a plateau around 3 g H₂O (g dry weight)^-1 in early winter when LT₅₀ was still decteasing, and then gradually increased under snow cover. A significant negative correlation was found between mean minimum air temperatures and freezing tolerance prior to the establishment of continuous snow cover. In contrast, a positive association between mean minimum temperatures and crown tissue water content was significant only when air temperatures were above 0°C, as water content did not decrease further at sub-zero temperatures. Seasonal changes in T₁ were closely related to changes in freezing tolerance. T₁ decreased until January even though water content stopped decreasing. Further tests on 15 field-grown varieties confirmed a strong negative association between freezing tolerance and T₁. The results suggest that cold hardening is comprised of two stages, with the transition occurring at ca 0°C. Development of hardiness was related to (1) a reduction in water content in the first stage (at minimum temperatures > 0°C), and (2) a change in physical state of water without much reduction in water content in the second stage. Varietal differences in hardiness thus arise due to changes in both water content and physical state of water.     

植物抗寒及其基因表达研究进展

植物经过逐渐降低的温度从而提高抗寒能力 ,这个过程被人们称为低温驯化。植物低温驯化过程是一个复杂的生理、生化和能量代谢变化过程 ,这些变化主要包括膜系统的稳定性、可溶性蛋白的积累和小分子渗透物质 ,比如脯氨酸、糖等 ,这些变化中的一些是植物抗寒必需的 ,而另外一些变化不是必需的。主要对冷害和低温生理生化变化、低温诱导表达基因的功能和作用、低温驯化的调节机制及其信号转导方面进行了综述。通过差别筛选 c DNA文库的方法已经鉴定了许多低温诱导表达、进而提高植物抗寒能力的基因 ,其中有脱水素、COR基因和 CBF1转录因子等。低温信号的感受、转导和调节表达是低温驯化的关键环节 ,低温信号的转导过程与干旱胁迫之间具有一定的交叉 ,这为利用 ABA等来提高植物抗寒能力成为可能 ,相信不久的将来人们可以通过提高植物抗寒能力从而增加经济产量成为现实。     

Electrical resistance changes during exposure to low temperature measure chilling and freezing tolerance in olive tree (Olea europaea L.) plants

Electrical resistance changes in different organs of four olive tree (Olea europaea L.) varieties, characterized by different tolerance to chilling and freezing, were examined, during exposure to low temperature. Apparent critical temperatures (CT) and freezing temperatures (T_fr) were identified on the basis of the electrical resistance changes. Both temperatures were lower for the more chilling‐tolerant genotypes. From the apparent critical temperatures, the absolute critical temperature (CT_abs) and the time delay of the chilling signal transduction process were calculated. In shoots, CT_abs varied from 8·8 °C for Ascolana (chilling‐tolerant variety) to 13·6 °C for Coratina (chilling‐sensitive variety). The magnitude of the transduction time was very similar (about 2 min) for the three genotypes that are more sensitive to chilling, whereas it was significantly higher (about 3 min) for the most tolerant genotype. Different freezing temperatures were observed for different organs. It would appear from this experiment that the order of sensitivity is roots > leaves > shoots > vegetative buds. Accord was found between the absolute critical temperature of electrical resistance and the critical temperature of membrane potential. The occurrence of electrical resistance changes in the tissues of the olive trees exposed to low temperature suggests the use of this experimental procedure as a quick, easy and non‐destructive tool to screen plant tissues for chilling tolerance. The strong dependence of the electrical resistance on low temperature, and the critical temperature of around 10 °C, can yield interesting information about the lowest thermal limits for the continuation of normal physiological processes and therefore about the adaptability of plants to particular environments.     

Acquisition of freezing tolerance in early autumn and seasonal changes in gall water content influence inoculative freezing of gall fly larvae,Eurosta solidaginis (Diptera,Tephritidae)

We examined seasonal changes in freeze tolerance and the susceptibility of larvae of the gall fly, Eurosta solidaginis to inoculative freezing within the goldenrod gall (Solidago sp.). In late September, when the water content of the galls was high (approximately 55%), more than half of the larvae froze within their galls when held at -2.5 degrees C for 24 h, and nearly all larvae froze at -4 or -6 degrees C. At this time, most larvae survived freezing at > or = -4 degrees C. By October plants had senesced, and their water content had decreased to 33%. Correspondingly, the number of larvae that froze by inoculation at -4 and -6 degrees C also decreased, however the proportion of larvae that survived freezing increased markedly. Gall water content reached its lowest value (10%) in November, when few larvae froze during exposure to subzero temperatures > or = -6 degrees C. In winter, rain and melting snow transiently increased gall water content to values as high as 64% causing many larvae to freeze when exposed to temperatures as high as -4 degrees C. However, in the absence of precipitation, gall tissues dried and, as before, larvae were not likely to freeze by inoculation. Consequently, in nature larvae freeze earlier in the autumn and/or at higher temperatures than would be predicted based on the temperature of crystallization (T(c)) of isolated larvae. However, even in early September when environmental temperatures are relatively high, larvae exhibited limited levels of freezing tolerance sufficient to protect them if they did freeze.     

Photosynthesis in cold acclimated leaves of plants with various degrees of freezing tolerance

The objective of this study was to compare the photosynthetic changes during cold acclimation in various plant types able to acquire different degrees of freezing tolerance. Four herbaceous and six woody plants were hardened under natural or artificial conditions and – after determination of their frost resistance (LT₅₀) – the net photosynthetic rate at an ambient CO₂ of 33 Pa (P_n33), the dependencies of P_n to light and to CO₂ and the room temperature chlorophyll a fluorescence were recorded under optimal conditions. Herbaceous plants acquired freezing tolerances to temperatures between ?10 and ?15°C when hardened at temperatures around 0°C. Most leaves fully developed prior to frost hardening exhibited typical symptoms of senescence after frost hardening. In non-senescing leaves P_n33 was reduced by 15 to 50% mainly due to a reduced stomatal conductance. After hardening at temperatures around ?10°C Brassica survived down to ?24°C, but P_n33 was almost abolished as a result of disturbances in the chloroplasts. After transferring the plants to 20/15°C P_n33 recovered completely within a few days. Woody plants hardened at temperatures around 0°C tolerated – 15 to ?36°C: P_n33 was reduced by 25 to 60% and hardly recovered at 20/15°C. Hardening at ?10°C induced a tolerance of ?32 to n33 was almost totally blocked, but at 20/15°C it returned to the values of the plants hardened at 0°C within a few days. In woody plants disturbances were invariably localized in the chloroplasts. Thus, conifers, and especially Pinus cembra, can survive much lower temperatures than herbaceous plants and, at the same level of freezing tolerance, exhibit appreciably less restriction in relative P_n33.