植物抗寒机理研究进展

本文综合概述了国内外有关植物抗寒机理研究的动态,主要讨论了植物抗寒性与细胞膜系、酶系多态性及抗寒基因表达与调控之间的相关性。此外,亦提出了有关植物抗寒机制研究领域值得深入研讨的问题。     

植物抗寒机理进展

本文综合概述阵内外有关植物抗寒机理研究的动态。主要讨论了植物抗寒性与细胞膜系、酶系多态笥及抗寒基因表达与调控之间的相关性。此外,亦提出了有关植物抗寒机制研究领域值得深入研讨的问题。     

植物抗寒性与抗寒基因的表达和调控

综合概述了国内外有关植物抗寒机理的研究动态,主要讨论了抗寒基因的表达与调控在植物抗寒性中的反应。此外,亦提出了有关植物抗寒机制研究领域值得深入研讨的问题。     

植物抗寒冻基因工程研究进展

低温寒害是限制农作物产量和分布的一种全球性的自然灾害,提高农作物的抗寒性具有重要意义。目前随着植物寒害机理、抗寒冻和冷驯化分子机理的深入发展,已研究发现了多种抗寒基因,包括各种抗寒调控基因和各种抗寒功能基因,从而使植物抗寒冻基因工程的研究与应用得到了广泛开展,以期最终有效地提高农作物的抗寒性,增加农业产量。本文综合概述了国内外有关植物抗寒冻基因工程的最新研究方向、进展及成就,并提出了此领域尚存在的一些问题及其前景展望。     

植物抗寒冻基因工程研究进展

低湿寒害是限制农作物产量和分布的一种全球性的自然灾害。提高农作物的抗寒性具有重要意义。目前随着植物寒害机理、抗寒冻和冷驯化分子机理的深入发展,已研究发现了多种抗寒基因,包括各种抗寒调控基因和各种抗寒功能基因,从而使植物抗寒冻基因工程的研究与应用得以了广泛开展,以期最终有效地提高农作物的抗寒性,增加农业产量。本文综合概述了国内外有关植物抗寒冻基因工程的最新研究方向、进展及成就,并提出了此领域尚存在的一些问题及其前景展望。     

植物抗寒机理研究的新进展

近年来,在植物抗寒机理研究方面取得不少新进展,大体有以下几个方面:(1)确证细胞的膜体系与植物抗寒性存在密切关系;(2)抗寒植物避免细胞内结冰机制的一些新发现;(3)稳定越冬植物抗寒力机制的揭示;(4)关于抗寒特异性蛋白质研究的开展.现将有关文献资料分述如下.     

植物抗寒基因工程研究进展

温度是影响植物分布、产量及品质的重要环境因素,提高植物抗寒性对农业生产具有重要的意义.近年来,随着基因工程的发展,对植物的抗寒机理进行了深入的研究,并克隆了许多与抗寒相关的基因.本文从膜稳定性、抗氧化酶活性、抗冻蛋白、低温信号转录因子和渗透调节物质等方面对植物耐冷性基因工程研究进展进行了分析、归纳与总结,旨在为植物抗寒机理研究及植物抗寒育种提供参考.     

提高植物抗寒性的机理研究进展

低温胁迫是世界范围内影响植物产量和品质的主要非生物胁迫.植物抗寒生理生态研究是比较活跃和发展很快的领域.文章综述了提高植物抗寒性机理的研究进展.大量科学研究和生产实践表明,气象因素与植物自身因素是影响植物抗寒性的关键因素,前者主要是温度、光周期和水分,后者主要是植物的遗传学基础、生长时期、发育水平以及低温胁迫下细胞的抗氧化能力.保证植物抗寒基因充分表达对提高植物抗寒性有重要意义.植物抗寒性的遗传机制与调控主要通过5条路径实现:丰富多样的植物低温诱导蛋白,低温转录因子DREB/CBF可同时调控多个植物低温诱导基因的表达,DREB/CBF与辅助因子相互作用调控下游基因表达,Ca2+、ABA及蛋白质磷酸化上游调控低温诱导基因表达,以及不饱和脂肪酸酶基因的表达.基因工程改良植物抗寒性已获重要进展,但距产业化尚有许多开创性的工作要做,目前主要通过导入抗寒调控基因和抗寒功能基因而实现,后者主要是导入抗渗透胁迫相关基因、抗冻蛋白基因、脂肪酸去饱和代谢关键酶基因、SOD等抗氧化系统的基因以及与植物激素调节有关的基因.农林技术对提高植物抗寒性有重大实用价值,其中的不少技术蕴涵着深刻的科学机理,重点评述了抗寒育种、抗砧嫁接、抗寒锻炼、水肥耦合及化学诱导五大技术提高植物抗寒性的作用机理.展望了提高植物抗寒性的研究.     

植物抗寒基因工程研究进展

本文评述了近年来有关植物抗寒生理及分子生物学方面的研究进展,对冷诱导基因的功能,诱导调控以及抗寒转基因策略等做了总结。     

植物抗寒基因工程研究进展

本评述了近年来有关植物抗寒生理及分子生物学方面的研究进展,对冷诱导基因的功能,诱导调控以及抗寒转基因策略等做一总结。     

Effect of temperature on proton efflux from isolated chloroplast thylakoids

Temperature-induced changes in the decay of the light-induced proton gradient of chloroplast thylakoids isolated from chilling-resistant and chilling-sensitive plants have been examined. In the presence of N-methylphenazonium methosulfate, the thylakoids isolated from chilling-resistant barley (cv. Kanby) and pea (cv. Alaska) and chilling-sensitive mung bean (cv. Berken) plants showed temperature-induced changes at approximately 8.6, 13.3, and 14.0 C, respectively. Barley thylakoids assayed in the presence of sodium thiocyanate also showed a change at 8.6 C, whereas with no addition or upon the inclusion of both N-methylphenazonium methosulfate and sodium thiocyanate the change occurred at approximately 11.5 C.     

Temperature-induced Changes in Hill Activity of Chloroplasts Isolated from Chilling-sensitive and Chilling-resistant Plants

The effect of temperature on Hill activity has been compared in chilling-sensitive and chilling-resistant plants. The Arrhenius activation energy (Ea) for the photoreduction of 2,6-dichlorophenolindophenol by chloroplasts isolated from two chilling-sensitive plants, mung bean (Vigna radiata L. var. Mungo) and maize (Zea mays L. cv. PX 616), increased at low temperatures, below 17 C for mung bean and below 11 C for maize. However, the Ea for this reaction in pea (Pisum sativum L. cv. Massay Gem), a chilling-resistant plant, likewise increased at temperatures below 14 C. A second change in Ea occurred at higher temperatures. The Ea decreased above about 28 C for mung bean, 30 C for maize, and 25 C for pea. At temperatures approaching 40 C, thermal inactivation of Hill activity occurred. These results, when taken together with previous results obtained with the chilling-resistant plant barley, indicate that chloroplasts from both chilling-sensitive and chilling-resistant plants can undergo a change in chloroplast membrane activity at low temperatures above freezing and that the presence of such a change in chloroplast membranes is not necessarily correlated with chilling sensitivity.     

Roles of the plasma membrane and the cell wall in the responses of plant cells to freezing

In an effort to clarify the responses of a wide range of plant cells to freezing, we examined the responses to freezing of the cells of chilling-sensitive and chilling-resistant tropical and subtropical plants. Among the cells of the plants that we examined, those of African violet ( Saintpaulia grotei Engl.) leaves were most chilling-sensitive, those of hypocotyls in mungbean [ Vigna radiata (L.) R. Wilcz.] seedlings were moderately chilling-sensitive, and those of orchid [ Paphiopedilum insigne (Wallich ex Lindl.) Pfitz.] leaves were chilling-resistant, when all were chilled at -2 degrees C. By contrast, all these plant cells were freezing-sensitive and suffered extensive damage when they were frozen at -2 degrees C. Cryo-scanning electron microscopy (Cryo-SEM) confirmed that, upon chilling at -2 degrees C, both chilling-sensitive and chilling-resistant plant cells were supercooled. Upon freezing at -2 degrees C, by contrast, intracellular freezing occurred in Saintpaulia leaf cells, frost plasmolysis followed by intracellular freezing occurred in mungbean seedling cells, and extracellular freezing (cytorrhysis) occurred in orchid leaf cells. We postulate that chilling-related destabilization of membranes might result in the loss of the ability of the plasma membrane to act as a barrier against the propagation of extracellular ice in chilling-sensitive plant cells. We also examined the role of cell walls in the response to freezing using cells in which the plasma membrane had been disrupted by repeated freezing and thawing. In chilling-sensitive Saintpaulia and mungbean cells, the cells with a disrupted plasma membrane responded to freezing at -2 degrees C by intracellular freezing. By contrast, in chilling-resistant orchid cells, as well as in other cells of chilling-resistant and freezing-resistant plant tissues, including leaves of orchard grass ( Dactylis glomerata L.), leaves of Arabidopsis thaliana (L.) Heynh. and cortical tissues of mulberry ( Morus bombycis Koids.), cells with a disrupted plasma membrane responded to freezing by extracellular freezing. Our results indicate that, in the chilling-sensitive plants cells that we examined, not only the plasma membrane but also the cell wall lacked the ability to serve as a barrier against the propagation of extracellular ice, whereas in the chilling-resistant plant cells that we examined, not only the plasma membrane but also the cell wall acted as a barrier against the propagation of extracellular ice. It appears, therefore, that not only the plasma membrane but also the cell wall greatly influences the freezing behavior of plant cells.     

不同抗冷性水稻中编码甘油-3-磷酸转酰酶的部分cDNA的序列比较研究

采用ＲＴ－ＰＣＲ技术,以根据国外报道的几种双子叶植物的甘油－３－磷酸转酰酶的桎保守的氨基酸而设计的简并引物作为扩增引物,从不同抗冷性水稻品种中均扩增到一段约３１５ｂｐ的ｃＤＮＡ片段。测序结果表明它们都是编码ＧＰＡＴ的部分ｃＤＮＡ含有３１５个核苷酸,编码１０５个氨基酸。比产它们之间的核苷酸及推导氨基酸序列,发现有一定差异,且抗冷性相差越大的品种间差异越大。抗冷性的差异可能与脯氨枝的替换有关。     

Sulfoquinovosyl Diacylglycerols in Chilling-Sensitive and Chilling-Resistant Plants

Sulfoquinovosyl diacylglycerols were isolated from leaves ofchilling-sensitive and chilling-resistant plants. They werethen fractionated by argentation thin-layer chromatography onsilica gel according to the degree of unsaturation of theirfatty acids, and the molecular species compositions of the lipidswere determined from the fatty acid compositions of the separatedfractions. The results suggest that the saturated molecularspecies of sulfoquinovosyl diacylglycerol are not directly correlatedwith the chilling sensitivity of plants. (Received June 25, 1984; Accepted August 2, 1984)     

The effect of temperature of the rate of photosynthetic electron transfer in chloroplasts of chilling-sensitive and chilling-resistant plants

1. Photochemical activities as a function of temperature have been compared in chloroplasts isolated from chilling-sensitive (below approximately 12 °C) and chilling-resistant plants.2. An Arrhenius plot of the photoreduction of NADP⁺ from water by chloroplasts isolated from tomato (Lycopersicon esculentum var. Gross Lisse), a chilling-sensitive plant, shows a change in slope at about 12 °C. Between 25 and 14 °C the activation energy for this reaction is 8.3 kcal·mole^?1. Between 11 and 3 °C the activation energy increases to 22 kcal·mole^?1. Photoreduction of NADP⁺ by chloroplasts from another chilling-sensitive plant, bean (Phaseolus vulgaris var. brown beauty), shows an increase in activation energy from 5.9 to 17.5 kcal·mole^?1 below about 12 °C.3. The photoreduction of NADP⁺ by chloroplasts isolated from two chilling-resistant plants, lettuce (Lactuca sativa var. winter lake) and pea (Pisum sativum var. greenfeast), shows constant activation energies of 5.4 and 8.0 kcal·mole^?1, respectively, over the temperature range 3–25 °C.4. The effect of temperature on photosynthetic electron transfer in the chloroplasts of chilling-sensitive plants is localized in Photosystem I region of photosynthesis. Both the photoreduction of NADP⁺ from reduced 2,6-dichlorophenol-indophenol and the ferredoxin-NADP⁺ reductase (EC 1.6.99.4) activity of choroplasts of chilling-sensitive plants show increases in activation energies at approximately 12 °C whereas Photosystem II activity of chloroplasts of chilling-sensitive plants shows a constant activation energy over the temperature range 3–25 °C. The photoreduction of Diquat (1,1′-ethylene-2,2′-dipyridylium dibromide) from water by bean chloroplasts, however, does not show a change in activation energy over the same temperature range. The activation energies of each of these reactions in chilling-resistant plants is constant between 3 and 25 °C.5. The effect of temperature on the activation energy of these reactions in chloroplasts from chilling-sensitive plants is reversible.6. In chilling-sensitive plants, the increased activation energies below approximately 12 °C, with consequent decreased rates of reaction for the photoreduction of NADP⁺, would result in impaired photosynthetic activity at chilling temperatures. This could explain the changes in chloroplast structure and function when chilling-sensitive plants are exposed to chilling temperatures.     

低温光抑制恢复过程中黄瓜叶片PSⅡ活性及其电子传递对PSⅠ的影响

以“津春4号”黄瓜为试材,通过测定黄瓜叶片叶绿素荧光快速诱导动力学曲线和对820 nm光的吸收曲线,结合叶绿素荧光淬灭分析,研究低温光胁迫(4℃,200 μmol·m-2·s-1)6 h后,黄瓜叶片在常温(25℃)不同光强(0、15、200μmol·m-2·s-1)下PS Ⅰ和PS Ⅱ活性的恢复,以及恢复过程中PS Ⅰ与PS Ⅱ的相互作用.结果表明:低温光胁迫6h后,PS Ⅰ和PS Ⅱ发生不同程度的光抑制.在常温恢复阶段,PS Ⅱ活性快速恢复且对光强不敏感;PS Ⅰ活性在弱光下(15 μmol·m-2·s-1)快速恢复,在较强光(200 μmol·m-2·s-1)下恢复较慢.在低温光抑制恢复过程中,常温下PS Ⅱ活性恢复较快可能导致PS Ⅱ向PS Ⅰ的线性电子传递过快,进而抑制PS Ⅰ的活性恢复.因此,在进行黄瓜抗冷性育种时,不应该仅追求较高的PS Ⅱ抗性和较快的PS Ⅱ恢复速度,还应该注意两个光系统活性的协调.在生产中,应当在低温逆境发生及其之后较长一段时间内采取措施降低叶表面光照强度,以利于对植株光合机构的保护和光合活性的恢复.     

Temperature-dependent phase behavior of phosphatidylglycerols from chilling-sensitive and chilling-resistant plants

Seven major lipid classes were isolated from leaves of chilling-sensitive and chilling-resistant plants, and the temperature-dependent phase behaviors of their aqueous dispersions were studied by a fluorescence polarization method using trans-parinaric acid and its methyl ester. Phosphatidylglycerols from the chilling-sensitive plants went from the liquid crystalline state into the phase separation state at about 30°C in 100 mm NaCl and at about 40°C in 5 mm MgCl₂. In contrast, phosphatidylglycerols from the chilling-resistant plants went into the phase separation state at a much lower temperature. The other classes of lipids remained in the liquid crystalline state at all temperatures between 5°C and 40°C regardless of the chilling sensitivity of the plants, except sulfoquinovosyl diacylglycerol from sponge cucumber in which phase separation seemed to begin at about 15°C. Compositions and positional distributions of fatty acids of the lipids suggest that the phosphatidylglycerols from the chilling-sensitive plants, but no other lipids, contained large proportions of molecular species which undergo phase transition at room temperature or above. The thermotropic phase behaviors and the fatty acid compositions suggest that, among the major lipid classes from leaves of the chilling-sensitive plants, only phosphatidylglycerol can induce a phase transition. Since a major part of this lipid in leaves originates from the chloroplasts, phase transition probably occurs in the chloroplast membranes.     

Molecular Species Composition of Phosphatidylglycerols from Chilling-Sensitive and Chilling-Resistant Plants

The molecular species of phosphatidylglycerols from leaves of9 species of chilling-sensitive plants and 12 species of chilling-resistantplants were analyzed by gas-liquid chromatography and gas chromatography-massspectrometry. The sum of the contents of the dipalmitoyl plusthe 1-palmitoyl-2-(trans-3-hexadecenoyl) species of phosphatidylglycerolranged from 3 to 19% of the total of this lipid in the chilling-resistantplants, and from 26 to 65% in the chilling-sensitive plants.These findings suggest that these two molecular species of phosphatidylglycerolsare closely associated with the chilling sensitivity of theplants. The biochemical difference between the chilling-sensitiveand the chilling-resistant plants is discussed in terms of theactivities of enzymes involved in phosphatidylglycerol biosynthesisin the chloroplasts. (Received August 19, 1982; Accepted November 19, 1982)     

A Comparison of the Effects of Chilling on Thylakoid Electron Transfer in Pea (Pisum sativum L.) and Cucumber (Cucumis sativus L.)

Experiments comparing the photosynthetic responses of a chilling-resistant species (Pisum sativum L. cv Alaska) and a chilling-sensitive species (Cucumis sativus L. cv Ashley) have shown that cucumber photosynthesis is adversely affected by chilling temperatures in the light, while pea photosynthesis is not inhibited by chilling in the light. To further investigate the site of the differential response of these two species to chilling stress, thylakoid membranes were isolated under various conditions and rates of photosynthetic electron transfer were determined. Preliminary experiments revealed that the integrity of cucumber thylakoids from 25°C-grown plants was affected by the isolation temperature; cucumber thylakoids isolated at 5°C in 400 millimolar NaCl were uncoupled, while thylakoids isolated at room temperature in 400 millimolar NaCl were coupled, as determined by addition of gramicidin. The concentration of NaCl in the homogenization buffer was found to be a critical factor in the uncoupling of cucumber thylakoids at 5°C. In contrast, pea thylakoid membranes were not influenced by isolation temperatures or NaCl concentrations. In a second set of experiments, thylakoid membranes were isolated from pea and cucumber plants at successive intervals during a whole-plant light period chilling stress (5°C). During wholeplant chilling, thylakoids isolated from cucumber plants chilled in the light were uncoupled even when the membranes were isolated at warm temperatures. Pea thylakoids were not uncoupled by the whole-plant chilling treatment. The difference in integrity of thylakoid membrane coupling following chilling in the light demonstrates a fundamental difference in photosynthetic function between these two species that may have some bearing on why pea is a chilling-resistant plant and cucumber is a chilling-sensitive plant.