不同胁迫预处理提高水稻幼苗抗寒性期间蛋白质的变化

水稻(Oryza sativa L.)幼苗经盐、热激和冷三种不同胁迫预处理均提高了幼苗的抗寒性。与未预处理苗相比,在处理后、低温伤害后和常温下恢复2d的三个时期,不同胁迫预处理苗的可溶性和热不稳定蛋白含量变化趋势甚为相似,但热稳定蛋白含量变化则各有异同。SDS-PAGE图谱分析显示,不同胁迫预处理提高水稻幼苗抗寒性时,其可溶性蛋白、热稳定和热不稳定蛋白组成变化亦各有异同。除诱导出共有的新多肽外,还各自诱导出特有的新多肽。结果表明,植物对不同胁迫的交叉适应存在一定的共同机理,但亦可看出植物对同一种环境胁迫似乎不是以同一的机理去适应。     

冷和盐预处理提高水稻幼苗抗寒性期间细胞Ca^2＋—ATP酶活性的变化

冷和盐预处理显著提高水稻（OryzasatwaL．）幼苗的抗寒性,但两预处理诱导提高的抗寒性可为钙的螫合剂EGTA和钙调素的抑制剂CPZ所抑制。冷预处理有提高很质膜、液泡膜和叶片的叶绿体Ca2 ．ATPase活性和质膜Fe（CN）3-6还原活性的作用,其中对提高根质膜Ca2 －ATPase活性和Fe（CN）3-6还原活性的作用尤为显著。盐预处理对提高根质膜、幼叶叶绿体Ca2 ATP酶活性和质膜Fe（CN）3-6还原活性的作用类似于冷预处理。虽然盐预处理苗液泡膜Ca2 －ATPase活性有所下降,但其活性仍明显高于未预处理苗,表明在低温胁迫下,两预处理苗都有较强的维持Ca2 稳态能力。结果揭示,冷和盐预处理诱导水稻幼苗抗寒性的提高,可能均与在低温胁迫下两预处理能有效地维持或激活Ca@ －ATPase活性有关,两者有着类似的适应机制。     

盐和水分预处理对扶芳藤幼苗抗寒性的影响

测定了扶芳藤新品种‘红脉'经不同方式预处理后对幼苗抗寒性的影响,测定了膜脂过氧化水平、保护酶活性和渗透调节物质含量的变化.结果表明,与对照低温胁迫相比,0.1和0.3mol/L NaCl溶液预处理后再经低温胁迫处理,扶芳藤幼苗的膜脂过氧化水平较低、保护酶活性高、渗透调节物质含量的累积较多,不同浓度的NaCl溶液预处理比不同浓度的PEG溶液预处理对提高扶芳藤的抗寒性更有效.通过实验证明了植物在不同的逆境中确实存在着交叉适应现象,植物对各种逆境胁迫有着许多相同的生理反应,对各种胁迫的适应性存在某些共同的机制.     

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。     

钙对萝卜幼苗抗寒性及某些生理指标的影响

研究结果表明钙能够提高萝卜幼苗的抗寒性,同时与抗寒性有关某些生理指标也受到影响。在低温胁迫后及恢复生长后处理组的萝卜幼苗的超氧岐化酶、过氧化物酶、抗坏血酸过氧化物酶、过氧化氢酶的活性都对照组高;在低温胁迫后处理组有脯氨酸及抗坏血酸的含量均比对照组高,但修复组萝卜幼苗的丙二醛含量却比对照组低。表明钙对萝卜幼苗抗寒性的影响可能与其防止膜脱过氧化有关。     

Ca2+预处理对热胁迫下辣椒叶肉细胞中Ca2+-ATP酶活性的影响

在常温下生长的辣椒(Capsicum annum L.)叶肉细胞中Ca2+-ATP酶主要分布于质膜、液泡膜上,叶绿体的基质和基粒片层上也有少量分布;在40℃下热胁迫不同的时间,酶活性逐渐下降,直至叶绿体超微结构解体.同样条件下,经过Ca2+预处理后,分布在上述细胞器膜或片层上的酶活性大大提高,表明Ca2+预处理对该酶活性具有激活作用;Ca2+预处理对热胁迫下的超微结构的完整性具有一定的保护作用,并且能使Ca2+-ATP酶在热胁迫下维持较高活性.结果表明,Ca2+预处理增强辣椒幼苗的抗热性,可能与其稳定细胞膜、从而使Ca2+-ATP酶在热胁迫下保持较高活性有一定关系.     

氧化胁迫对水稻幼苗抗冷力的影响

利用Ｈ２Ｏ２和甲基紫精（ＭＶ）对水稻幼苗作三种不同程度的氧化胁迫预处理。结果表明：轻度氧化胁迫预处理（１０ｕｍｏｌ／ＬＨ２Ｏ２或１０ｕｍｏｌ／ＬＭＶ处理４ｈ）提高了水稻幼苗的抗冷力,严重氧化胁迫预处理（１０ｕｍｏｌ／ＬＨ２Ｏ２或１０ｕｍｏｌ／ＬＭＶ分别处理１６ｈ和４０ｈ）则削弱水稻幼苗的抗冷力。氧化胁迫预处理刺激了水稻幼苗叶片抗氧化酶（ＳＯＤ,ＣＡＴ,ＰＯＸ和ＡＰＸ）的活性。经冷胁迫后,不同预处理苗的叶片抗氧化酶活性、膜脂过氧化和膜结构的变化趋势不同：轻度氧化胁迫预处理使幼苗仍保持较高的抗氧化酶活性,减轻了由冷胁迫引起的膜脂过氧化和细胞膜的渗漏程度,而严重氧化胁迫预处理则相反。因此,水稻幼苗对氧化胁迫感知并作出反应的机制（氧化应激机制）在水稻幼苗对低温反应和适应过程中起着很重要的调节作用。     

干旱胁迫下Ca2+·CaM信使系统对稻苗保护酶活性的影响

以氯丙嗪(CPZ)和LaCl3对水稻幼苗根际预处理,阻断其Ca2+·CaM信使系统传导后,研究了干旱胁迫下稻苗膜脂过氧化和保护酶活性的变化.结果表明干旱胁迫下,LaCl3和CPZ根际预处理显著加剧稻苗膜脂过氧化,加剧过氧化氢酶(CAT)活性和抗坏血酸过氧化物酶(APX)活性下降,对胁迫前期超氧化物岐化酶(SOD)和过氧化物酶(POD)活性无显著影响,且CPZ和LaCl3预处理加剧CAT和APX活性下降与加剧稻苗MDA含量积累分别呈极显著和显著正相关.这些结果暗示Ca2+·CaM信使系统可能主要通过调节或影响CAT和APX活性而调节稻苗的抗旱性.     

Ca^2＋预处理对热胁迫下辣椒叶肉细胞中Ca^2＋—ATP酶活性的影响

在常温下生长的辣椒（Capsicum annuum L.）叶肉细胞中Ca^2 －ATP酶主要分布于质膜、液泡膜上,叶绿体的基质和基粒片层上也有少量分布;在40℃下热胁迫不同的时间,酶活性逐渐下降,直到叶绿体超微结构解体。同样条件下,经过Ca^2 预处理后,分布在上述细胞器膜或片层上的酶活性大大提高,表明Ca^2 预处理对该活性具有激活作用;Ca^2 预处理对热胁迫下的超微结构的完整性具有一定的保护作用,并且能使Ca^2 －ATP酶在热胁迫下维持较高活性。结果表明,Ca^2 预处理增强辣椒幼苗的抗热性,可能与其稳定细胞膜、从而使Ca^2 －ATP酶在热迫下保护较高活性有一定关系。     

NaCl胁迫对栓皮栎幼苗生长及其生理响应

为了研究栓皮栎幼苗期对盐胁迫的生理耐受特性,选择2年生栓皮栎实生幼苗为材料,在盆栽条件下,设置NaCl盐分梯度(0%、0.4%、0.6%、0.8%),系统测定分析栓皮栎幼苗在不同盐胁迫梯度和胁迫时间下的形态生长指标以及保护酶活性、脯氨酸含量、根系活力等各项生理指标。结果显示:(1)随着盐胁迫程度的加剧,栓皮栎幼苗各部分器官鲜重和干重以及株高、基径和一级侧根长均先升高后降低,而主根长度逐渐增加,叶片数逐渐减少,并在重度胁迫下达到显著水平。(2)随着盐胁迫时间的延长,栓皮栎幼苗的超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性均呈先升高后降低的趋势,但变化时间和幅度不同;丙二醛(MDA)含量则日益增加,且胁迫度越大增加越显著。(3)栓皮栎幼苗叶片的渗透调节物质脯氨酸和可溶性蛋白含量在低盐胁迫下不断升高,高盐胁迫下先升高后降低;而可溶性糖的含量则随时间延长和胁迫加剧显著增加。(4)随NaCl浓度的提高,幼苗根系活力值先增后减,叶绿素a和叶绿素b含量均呈下降趋势。研究表明,在轻、中度NaCl胁迫下,栓皮栎幼苗通过提高保护酶活性、增加渗透调节物质等策略缓解盐胁迫伤害,表现出一定的耐盐潜力;而在高浓度盐分胁迫下幼苗受到伤害,自我调节能力降低,导致保护酶活性、可溶性蛋白含量和根系活力等下降。     

水稻幼苗冷锻炼过程中钙的效应

冷锻炼处理提高了水稻（Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．）幼苗叶片中抗氧化剂（还原型谷胱甘肽,ＧＳＨ;抗坏血酸,ＡｓＡ）含量和膜保护酶（超氧化物歧化酶,ＳＯＤ）的活性,同时也提高了可溶性蛋白质中热稳定蛋白的含量。ＣａＣｌ２ 浸种处理对上述冷锻炼的作用有加强的效果,且明显地提高了过氧化氢酶（ＣＡＴ）和过氧化物酶（ＰＯＤ）的活性。有无ＣａＣｌ２ 处理的冷锻炼处理均减轻冷胁迫引起的ＧＳＨ 及ＡｓＡ 含量、ＳＯＤ 活性及热稳定蛋白质含量的下降程度,有利于幼苗在恢复过程中ＧＳＨ、ＡｓＡ、ＣＡＴ、ＳＯＤ、ＰＯＤ及热稳定蛋白质水平迅速回升。结合ＣａＣｌ２ 处理的冷锻炼苗在冷胁迫恢复生长时增长迅速,且苗健壮浓绿,说明ＣａＣｌ２浸种对冷锻炼处理提高水稻幼苗的抗冷力有明显的促进作用,这与ＣａＣｌ２ 浸种结合冷锻炼能更有效的提高细胞膜保护能力有关     

冷锻炼和ABA诱导水稻幼苗提高抗冷性期间膜保护系统的变化

冷锻炼和ＡＢＡ处理提高了水稻幼苗叶绿体ＳＯＤ和ＧＲ活性及叶片抗氧化剂ＡｓＡ和ＧＳＨ的含量,降低了膜电解质泄漏,增强了幼苗的抗冷性．等电聚焦电泳分析表明,冷锻炼和ＡＢＡ处理苗叶绿体ＳＯＤ三条同工酶带和ＧＲ１、２、３和６同工酶带都有不同程度的增强．低温胁迫后,处理和未处理首的ＳＯＤ、ＧＲ活性和ＡＳＡ、ＧＳＨ含量均有所下降．但处理苗的水平仍维持在未处理苗之上．亚胺环已酮可抑制因冷锻炼和ＡＢＡ诱导增加的ＳＯＤ和ＧＲ活性,并使叶片电解质泄漏增大．本试验结果表明冷锻炼或ＡＢＡ诱导水稻幼苗抗冷性提高时,对防御活性氧的保护系统有类似的影响。     

Combined action of antioxidant defense system and osmolytes in chilling shock-induced chilling tolerance in Jatropha curcas seedlings

Low non-freezing temperature is one of the major environmental factors affecting growth, development and geographical distribution of chilling-sensitive plants, Jatropha curcas is considered as a sustainable energy plants with great potential for biodiesel production. In this study, chilling shock at 5 °C followed by recovery at 26 °C for 4 h significantly improved survival percentage of J. curcas seedlings under chilling stress at 1 °C. In addition, chilling shock could obviously enhance the activities of antioxidant enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT) and glutathione reductase (GR), and the levels of antioxidants ascorbic acid (AsA) and glutathione (GSH), as well as the contents of osmolytes proline and betaine in leaves of seedlings of J. curcas compared with the control without chilling shock. During the process of recovery, GR activity, AsA, GSH, proline and betaine contents sequentially increased, whereas SOD, APX and CAT activities gradually decreased, but they markedly maintained higher activities than those of control. Under chilling stress, activities of SOD, APX, CAT, GR and GPX, and contents of AsA, GSH, proline and betaine, as well as the ratio of the reduced antioxidants to total antioxidants [AsA/(AsA + DHA) and GSH/(GSH + GSSG)] in the shocked and non-shock seedlings all dropped, but shocked seedlings sustained significantly higher antioxidant enzyme activity, antioxidant and osmolyte contents, as well as ratio of reduced antioxidants to total antioxidants from beginning to end compared with control. These results indicated that the chilling shock followed by recovery could improve chilling tolerance of seedlings in J. curcas, and antioxidant enzymes and osmolytes play important role in the acquisition of chilling tolerance.     

Heat shock-mediated APX gene expression and protection against chilling injury in rice seedlings

Rice (Oryza sativa L.) seedlings, when kept at 42 degrees C for 24 h before being kept at 5 degrees C for 7 d, did not develop chilling injury. Chilling resistance was enhanced in parallel with the period of heat-treatment. The level of APX activity was higher in heated seedlings whereas CAT activity was decreased by heat stress. There was no significant difference in SOD activity between heated and unheated seedlings. The elevated activity of APX was sustained after 7 d of chilling. The cytosolic APX gene expression in response to high and low temperature was analysed with an APXa gene probe. APXa mRNA levels increased within 1 h after seedlings were exposed to 42 degrees C. Elevated APXa mRNA levels could also be detected after 24 h of heating. The APXa mRNA level in preheated seedlings was still higher than unheated seedlings under cold stress. The promoter of the APXa gene was cloned from rice genomic DNA by TAIL-PCR, and characterized by DNA sequencing. The promoter had a minimal heat shock factor-binding motif, 5'-nGAAnnTTCn-3', located in the 81 bp upstream of the TATA box. Heat shock induction of the APXa gene could be a possible cause of reduced chilling injury in rice seedlings.     

热激对水稻幼苗耐冷性及热激蛋白合成的诱导

萌发的水稻种子经42℃热激处理后其幼苗的耐冷性明显增强,膜伤害程度降低,脯氨酸含量增加,超氧物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)活性和抗氧化物质抗坏血酸含量增加,而膜脂过氧化的关键酶脂氧合酶(LOX)活性及其产物丙二醛(MDA)含量下降.并且热激诱导萌发的水稻胚合成78、70、64、60、46、38、24、17、16kD的热激蛋白(HSP),其中属于HSP70的内质网结合蛋白(BiP)的合成与水稻幼苗耐寒性的提高有关.     

Effects of Temperature Acclimation Pretreatment on the Ultrastructure of Mesophyll Cells in Young Grape Plants （Vitis vinifera L. cv. Jingxiu） Under Cross-Temperature Stresses

Leaves from annual young grape plants （Vitis vinifera L. cv. Jingxiu） were used as experimental materials. The ultrastructural characteristics of mesophyll cells in chilling-treated plants after heat acclimation （HA） and in heat-treated plants after cold acclimation （CA） were observed and compared using transmission electron microscopy. The results showed that slight injury appeared in the ultrastructure of mesophyll cells after either HA （38℃ for 10 h） or CA （8℃ for 2.5 d）, but the tolerance to subsequent extreme temperature stress was remarkably improved by HA or CA pretreatment. The increases in membrane permeability and malondialdehyde concentration under chilling （0℃） or heat （45℃） stress were markedly inhibited by HA or CA pretreatment. The mesophyll cells of plants not pretreated with HA were markedly damaged following chilling stress. The chloroplasts appeared irregular in shape, the arrangement of the stroma lamellae was disordered, and no starch granules were present. The cristae of the mitochondria were disrupted and became empty. The nucleus became irregular in shape and the nuclear membrane was digested. In contrast, the mesophyll cells of HA-pretreated plants maintained an intact ultrastructure under chilling stress. The mesophyll cells of control plants were also severely damaged under heat stress. The chloroplast became round in shape, the stroma lamellae became swollen, and the contents of vacuoles formed clumps. In the case of mitochondria of control plants subjected to heat stress, the outer envelope was digested and the cristae were disrupted and became many small vesicles. Compared with cellular organelles in control plants, those in CA plant cells always maintained an integrated state during whole heat stress, except for the chloroplasts, which became round in shape after 10 h heat stress. From these data, we suggest that the stability of mesophyll cells under chilling stress can be increased by HA pretreatment. Similarly, CA pretreatment can protect chloroplasts, mitochondria, and the nucleus against subsequent heat stress; thus, the thermoresistance of grape seedlings was improved. The results obtained in the present study are the first, to our knowledge, to offered cytological evidence of cross-adaptation to temperature stresses in grape plants.     

Thermotolerance and antioxidant response induced by heat acclimation in <Emphasis Type="Italic">Freesia</Emphasis> seedlings

Followed a heat acclimation pretreatment, seedlings of Freesia hybrida ‘Shangnong Jinghuanghou’ were exposed to heat stress at 38°C for 6 h treatment and then recovered at 22°C for 72 h to study the impact of heat acclimation (30°C) on thermotolerance under heat stress. The results showed that the pretreated seedlings performed better under heat stress than control. Heat acclimation could slow down the decrease of chlorophyll contents under heat stress and recover better. Higher levels of soluble sugar and proline and slight lower level of soluble protein were observed in pretreated seedlings. After recovery, similar levels of proline and soluble protein were maintained in all seedlings. However, a higher level of soluble sugar was maintained in pretreated seedlings. MDA content and EL showed a stable level in pretreated seedlings while a significant increase in control, followed by a significant decrease after recovery. Significant different responses of SOD, POD, CAT, and APX activities were observed in pretreated seedlings and control. Heat acclimation led to higher activities of these enzymes and a significant response of antioxidant enzyme activities occurred in a time-dependent manner under heat stress. Exposure to high temperature caused a significant increase in SOD and APX activity, and much higher levels in SOD and APX activity were observed in pretreated seedlings compared to control during heat stress. A slight difference in change pattern of POD and CAT activity was presented. The highest activities of POD and CAT were observed at 4 and 6 h of heat stress in pretreated seedlings and control, respectively. After 72 h recovery, the activities of all tested enzymes decreased to similar levels in all seedlings.     

苯丙氨酸解氨酶在诱导黄瓜幼苗抗寒性中的作用

为了探讨苯丙氨酸解氨酶(PAL)在诱导黄瓜幼苗抗寒性中的作用,采用喷施特异抑制剂(AOPP)的方法控制PAL活性,测定幼苗抗寒性的变化.结果表明： 低温可以诱导黄瓜幼苗叶片中PAL的基因表达和活性升高;喷施AOPP显著抑制了叶片中PAL活性,减少了酚类和类黄酮物质的积累.低温对黄瓜幼苗造成显著伤害,AOPP预处理加剧了低温对幼苗的损伤,幼苗抗寒性降低.与对照相比,幼苗叶片中相对电解质渗漏率和丙二醛(MDA)含量显著升高,PSII最大光化学效率(F_v/F_m)降低,光化学猝灭参数Y(NO)升高,胁迫相关基因(PR1-1a、COR47、P5CS、HSP70)的诱导表达受到抑制.低温导致黄瓜幼苗叶片中H₂O₂积累,还原型抗坏血酸(AsA)含量降低,脱氢抗坏血酸(DHA)含量升高,AsA∶DHA减小;喷施AOPP的幼苗中抗氧化酶(过氧化氢酶CAT、抗坏血酸过氧化物酶APX)活性显著低于对照,H₂O₂过量积累,AsA∶DHA更低.施用H₂O₂清除剂可以有效缓解喷施AOPP引起的低温损伤加剧,而施用CAT抑制剂的幼苗对低温胁迫更敏感.表明低温诱导了PAL活性升高,促进了苯丙烷类次生代谢产物的合成,提高了胞内抗氧化酶活性,可有效清除活性氧分子,维持AsA氧化还原状态,缓解低温引起的光损伤和氧化损伤.