ABA and Low Temperature Induce Freezing Tolerance via Distinct Regulatory Pathways in Wheat

The role of ABA in the induction of freezing tolerance was investigatedin two wheat (T. aestivum L.) cultivars, Glenlea (spring var)and Fredrick (winter var). Exogenous application of ABA (5x10^–5M for 5 days at 24°C) increased the freezing tolerance ofintact plants by only 3°C (LT₅₀) in both cultivars. Maximalfreezing tolerance (LT₅₀ of –9°C for Glenlea and –17°Cfor Fredrick) could only be obtained with a low temperaturetreatment (6/2°C; day/night) for 40 days. These resultsshow that exogenously applied ABA cannot substitute for lowtemperature requirementto induce freezing tolerance in intactwheat plants. Furthermore, there was no increase in the endogenousABA level of wheat plants during low temperature acclimation,suggesting the absence of an essential role for ABA in the developmentof freezing tolerance in intact plants. On the other hand, ABAapplication (5x10^–5 M for 5 days at 24°C) to embryogenicwheat calli resulted in an increase of freezing tolerance similarto that achieved by low temperature. However, as in intact plants,there was no increase in the endogenous ABA level during lowtemperature acclimation of calli. These results indicate thatthe induction of freezing tolerance by low temperature is notassociated with an increase in ABA content. Using an antibodyspecific to a protein family associated with the developmentof freezing tolerance, we demonstrated that the induction offreezing tolerance by ABA in embryogenic wheat calli was correlatedwith the accumulation of a new 32 kDa protein. This proteinis specifically induced by ABA but shares a common antigenicitywith those induced by low temperature. These results suggestthat ABA induces freezing tolerance in wheat calli via a regulatorymechanism different from that of low temperature. (Received June 15, 1993; Accepted September 16, 1993)     

Cold-Induced Freezing Tolerance in Arabidopsis

Changes in the physiology of plant leaves are correlated with enhanced freezing tolerance and include accumulation of compatible solutes, changes in membrane composition and behavior, and altered gene expression. Some of these changes are required for enhanced freezing tolerance, whereas others are merely consequences of low temperature. In this study we demonstrated that a combination of cold and light is required for enhanced freezing tolerance in Arabidopsis leaves, and this combination is associated with the accumulation of soluble sugars and proline. Sugar accumulation was evident within 2 h after a shift to low temperature, which preceded measured changes in freezing tolerance. In contrast, significant freezing tolerance was attained before the accumulation of proline or major changes in the percentage of dry weight were detected. Many mRNAs also rapidly accumulated in response to low temperature. All of the cold-induced mRNAs that we examined accumulated at low temperature even in the absence of light, when there was no enhancement of freezing tolerance. Thus, the accumulation of these mRNAs is insufficient for cold-induced freezing tolerance.     

转拟南芥ICE1基因增强烟草抗寒性的研究

ICE1是CBF冷响应通道的上游转录调控因子,通过与CBF启动子中MYC顺式作用元件的结合激活CBF3基因表达.采用RT-PCR方法,从拟南芥获得AtICE1基因,将AtICE1导入pCAMBIA1301构建35S:AtICE1植物表达载体.通过根癌农杆菌GV3101,将AtICE1基因导人烟草,T1代植株经潮霉素抗性筛选,PCR、RT-PCR检测,结果表明AtICE1基因已经整合到烟草基因组中,并在转录水平表达;在正常生长条件下,转基因烟草与对照烟草的生长未见明显区别,而在瞬时低温冻害下,转基因烟草存活率明显高于对照烟草植株,说明Atl-CEI基因可以提高低温敏感作物的耐寒性.     

Temperature Drop as a Tool for Cold Tolerance Increment in Plants

The experiments were conducted with Cucumis sativus L., Triticum aestivum L., Brassica capitata, Solanum tuberosum, Zea mays L. and Pisum sativum L. Temperature drop increased cold resistance in all studied species. Level of cold resistance under the drop treatments was much higher than under the constant low temperature. It remained at a higher level for a longer period during de-acclimation. It is concluded that DROP technology is a good way for successful transplantation of greenhouse agricultural and bedding plants in early spring.     

高山植物的抗寒抗冻特性

综述了高山植物光合作用对冰冻低温胁迫的反应和适应特性,阐述了高山植物呼吸作用的低温适应特点;对高山植物抗冻机制作了详细分析和讨论。     

生菜种子低温处理后耐冻性和脂肪酸合成代谢的关系研究

以生菜(Lactuca sativa)种子为研究对象,通过不同时间的吸水处理分析其含水量变化,再通过程序降温处理,分析不同含水量种子发芽率的差异,以及脂肪酸合成有关基因(FAD2、FAD3、PPT、ELOVL)和冷调节基因ICE1的表达。结果表明,种子含水量随吸水时间增加而升高。程序降温至同样的低温冷冻条件下(-20℃、-22℃),吸水时间小于6 h的种子发芽率较高,而吸水8 h以上的种子发芽率显著降低。种子吸水8 h含水量处于饱和状态,在此状态下种子对低温较为敏感,说明含水量对种子耐冻性有影响。冷冻处理后生菜种子基因表达检测结果表明,脂肪酸去饱和酶基因(FAD2、FAD3)、蛋白质棕榈酰基硫脂酶相关基因(PPT）、长链脂肪酸延伸酶相关基因(ELOVL)的表达水平均随着种子含水量增加呈上升趋势,吸胀10 h的种子表达量最高,此时种子由于高含水量所受冷冻伤害最大。基因ICE1在冷冻处理种子中的表达也随着吸水时间增加而升高,在吸水10 h时种子中表达量到最高水平。综上,种子含水量越高,所受冷冻伤害越大。但种子在低温条件下具有一定的抗冷反应,可通过相关基因的过表达调控合成更多不饱和脂肪酸、抗冻蛋白等提高含水种子耐冻性。     

Betaine Improves Freezing Tolerance in Wheat

The accumulation of the osmolyte betaine was found to be correlatedwith the development of freezing tolerance (FT) of two wheatcultivars where it increases by about three fold during thecold acclimation period. Exogenous betaine application resultedin a large increase in total osmolality mostly due to betaineaccumulation. Plants that accumulated betaine are more tolerantto freezing stress since a four day exposure to 250 mM betaineresulted in a LT₅₀ of –8°C (in spring wheat Glenlea)and –9°C (in winter wheat Fredrick) compared to –3°C(Glenlea) and –4°C (Fredrick) for control non-exposedplants. Betaine treatment (250 mM) during cold acclimation increasedFT in an additive manner since the LT₅₀ reached –14°C(Glenlea) and –22°C (Fredrick) compared to –8°C(Glenlea) and –16°C (Fredrick) for plants that arecold acclimated in the absence of betaine. These results showthat betaine treatment can improve FT by more than 5°C inboth non-acclimated and cold-acclimated plants. The betainetreatment resulted in the induction of a subset of low temperatureresponsive genes, such as the wcor410, and wcor413, that arealso induced by salinity or drought stresses. In addition tothese genetic responses, betaine treatment was also able toimprove the tolerance to photoin-hibition of PSII and the steady-stateyield of electron transport over PSII in a manner that mimickedcold-acclimated plants. These data also suggest that betaineimproves FT by eliciting some of the genetic and physiologicalresponses associated with cold acclimation. (Received April 23, 1998; Accepted September 4, 1998)     

Enhanced Tolerance to Ascochyta Blight in Chickpea Plants via Low Temperature Acclimation

Russian Journal of Plant Physiology - Low temperature (LT) and Ascochyta blight are two major stresses in chickpea (Cicer arietinum L.) cultivation. After exposure to LT treatments (acclimation,...     

高温对3种攀缘植物叶绿素荧光特性及耐脱水特性的影响

利用植物叶绿素荧光技术,分析了高温对五叶地锦、胶东卫矛和南蛇藤的叶绿素荧光特性及高温脱水过程中叶片相对含水量(RWC)的变化,以明确攀缘植物的抗热特性.结果显示:(1)温度大于42℃时,南蛇藤F0升高的幅度和Fm下降程度均低于五叶地锦和胶东卫矛;54℃时,五叶地锦和胶东卫矛的F0与Fm近似相等,表明PSⅡ的反应中心完全关闭.(2)42℃时,南蛇藤、胶东卫矛和五叶地锦的PSⅡ最大光化学效率(Fv/Fm)显著降低,可逆变化的温度范围是24℃~42℃.(3)脱水末期(20.5 h),五叶地锦RWC仍可高达(57.7±5.0)%,高于胶东卫矛的(19.6±4.1)%和南蛇藤的(11.6±2.5)%,但Fv/Fm显著降低时所对应的RWC分别为(75.6±3.3)%、(30.4±3.6)%和(19.1±3.3)%.研究表明,五叶地锦热稳定性和水稳定性均低于南蛇藤和胶东卫矛,但五叶地锦的耐脱水能力较高;高温(42℃)脱水过程中Fv/Fm值的改变可能主要是由于Fm下降造成的,而不是由于F0变化所致;建议在水分可以得到保证的高温地区,选用胶东卫矛和南蛇藤,而在水分条件不好,但温度相对低的地区,选择五叶地锦进行垂直绿化.     

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。     

Drought and Salt Tolerance in Plants

Agricultural productivity worldwide is subject to increasing environmental constraints, particularly to drought and salinity due to their high magnitude of impact and wide distribution. Traditional breeding programs trying to improve abiotic stress tolerance have had some success, but are limited by the multigenic nature of the trait. Tolerant plants such as Craterostigma plantagenium, Mesembryanthemum crystallinum, Thellungiella halophila and other hardy plants could be valuable tools to dissect the extreme tolerance nature. In the last decade, Arabidopsis thaliana, a genetic model plant, has been extensively used for unravelling the molecular basis of stress tolerance. Arabidopsis also proved to be extremely important for assessing functions for individual stress-associated genes due to the availability of knock-out mutants and its amenability for genetic transformation. In this review, the responses of plants to salt and water stress are described, the regulatory circuits which allow plants to cope with stress are presented, and how the present knowledge can be applied to obtain tolerant plants is discussed.     

Freezing Tolerance of Shoot and Flower Primordia of Coniferous Buds by Extraorgan Freezing

Shoot and flower primordia of vegetative and flower buds ofextremely or very hardy conifers belonging to the subfamilyAbietoideae of the Pinaceae, survived between –40 and–70?C by extraorgan freezing, which differed greatly dependingupon species. The water in these organs gradually froze outwith decreasing temperatures when cooled very slowly, whichenabled these organs to survive %40?C or below. The same icesegregation in shoot and flower primordia by extraorgan freezingwas observed in most of the temperate conifers belonging toTaxaceae, Cephalotaxaceae, Taxodiaceae and Cuppressaceae, makingthem resistant to temperatures between –15 and –25?C.In these conifers, scales acted as an ice sink, unlike the conifersof Abietoideae. The rates of cooling and exosmosis of waterin the shoot or flower primordia, their size, and their abilityto tolerate freeze-dehydration or its related stress play animportant role in determining whether death is caused by freeze-dehydrationor intraorgan freezing. Even in very hardy conifers, low temperature exotherms fromfreezing within the shoot primordia appeared between –30and –35?C on the DTA profiles when cooled continuouslyunder laboratory conditions from 5?C to –50?C at 2 to5?C/h. Appearance of low temperature exotherms always resultedin death. However, in the coldest area of Hokkaido, where theair temperature cools down to –40?C or below nearly everyyear, such an intraorgan freezing seems seldom to occur, especiallyin natural stands. On the other hand, low temperatures below–25?C seldom occur in warm-temperate climates. Thus, itmay be considered that in both boreal and temperate coniferstheir shoot and flower primordia seem to tolerate freeze dehydrationby extraorgan freezing under natural conditions. ¹ Contribution No. 2431 from the rnstitute of Low TemperatureScience. (Received March 27, 1982; Accepted August 12, 1982)     

Maintenance of Membrane Fluidity during Development of Freezing Tolerance of Winter Wheat Seedlings

Fluidity of membrane lipids of shoot and root tissue and of chloroplasts from young wheat seedlings of contrasting freezing tolerance was investigated by measuring the motion and order parameters after spin labeling. A striking similarity was observed in membrane lipid fluidity of the five cultivars grown at 22 C. After cold hardening by growth at 2 C, a small change in membrane lipid fluidity was observed, but this was not correlated with the development of freezing tolerance, and there was no alteration in the transition temperature of membrane lipids. The results show that neither changes in membrane lipid fluidity nor transition temperature are a necessary feature of cold acclimation in wheat.     

转TaDREB基因提高芦荟抗低温特性的研究

以库拉索芦荟(Aloe vera L.)幼茎为外植体,利用农杆菌介导法将携带小麦功能基因TaDREB的表达载体pBIR1转化库拉索芦荟,在添加3.0mg/L BA、250mg/L Carbenicillin和15~25mg/L G418的MS培养基上诱导、筛选丛生芽,连续筛选几代后,将2.0~3.0cm高的抗性再生芽转移到1/2MS生根培养基上壮苗,共获得58株生长良好的抗性植株。根据标记基因npt Ⅱ及目的基因TaDREB的序列设计上游引物,对所有抗性植株进行PCR检测,共获得3株PCR阳性植株,结果表明目的基因的转化效率为0.5%。将转基因阳性植株置于4℃低温处理2周,20℃冷冻处理30min,发现对照植株发生严重冻害,而转基因植株生长良好,抗低温特性明显提高。对低温胁迫条件下培养14天的转基因植株SOD、POD酶活性进行分析,结果表明低温胁迫下转基因植株SOD、POD活性均呈降－升－升－升趋势变化,与非转基因植株的变化趋势降－升－降－降有所不同。进一步利用电导率法检测,结果转基因植株电导率的平均值为0.456,明显低于对照的0.685;说明转基因芦荟细胞膜的稳定性得到很好的改善,是其抗低温特性提高的内在生理基础。