植物抗寒性的生理生态学机制研究进展

综述植物在冷驯化过程中发生的一系列生理生化变化.环境对植物抗寒性的影响主要与光诱导、温湿度以及气候的变化有关.植物表面形成冰层会引起植物的无氧呼吸,导致植物受害;光抑制诱导活性氧的产生,从而导致植物光合系统的退化,抗寒能力下降,而短日照诱导植物休眠,有利于植物抗寒.光敏色素则被认为是启动冷驯化的光受体;植物通过冷驯化增加碳水化合物的积累及病原体相关蛋白的合成,以增强对低温病原体的抵抗能力;气候的变化使植物遭受了更大的冷伤害风险.微管最初遇冷时部分的解体可以有效诱导植物抗寒性;抗氧化酶活性增强,植物体内糖、脯氨酸、多胺等内含物含量上升.植物休眠状态中的生理变化(种子的休眠、芽的休眠)与ABA敏感性的差异有关.对植物抗寒性分子机制的研究表明:COR基因的表达对于植物抗寒性和冷驯化是十分关键的;与气候梯度有关的基因梯度的分布说明寒冷地区的树种更为抗寒;多表型性状的数量性状分析,为重要的农艺性状标记辅助选择(MAS)提供基础.对植物抗寒过程中的信号转导进行研究发现,Ca2 是低温下参与调节冷驯化应答机制中信号转导途径的重要的第二信使.未来植物抗寒领域的研究热点为信号转导和基因调节,低温抗性的遗传学和遗传应用及代谢组学,气候变化对于植物抗寒的影响等方面.     

一个新的柽柳类萌芽素基因ThGLP的结构及其表达分析

萌芽素和类萌芽素蛋白在不同植物的各个生长阶段和胁迫相关过程中起到不同的作用。本研究首次从刚毛柽柳cDNA文库中获得类萌芽素蛋白全长基因ThGLP,该基因编码225个氨基酸,含有植物萌芽素和类萌芽素蛋白的功能序列。通过进化树分析发现该基因屑于真正萌芽素亚家族。利用实时定量PCR方法研究了该基因在PEG、NaCl、低温、CdCl2和ABA胁迫下不同时间的表达模式。结果显示PEG、NaCl、低温、CdCl2和ABA处理均能诱导ThGLP基因在柽柳的根和叶中的表达。结果表明ThGLP在柽柳根和叶中表达,参与非生物胁迫应答并由ABA依赖的信号传导途径调控。     

CBF转录因子及其在林木抗寒研究中的应用前景

CBF转录因子是一类受低温诱导的反式作用因子,广泛存在于各种植物中。它可以与启动子中CRT/DRE顺式作用元件特异结合,激活其下游冷应答基因COR的表达,从而提高植物的抗寒性。自CBF1基因被发现以来,关于CBF基因结构及功能方面的研究已有很多报道,为提高植物的抗寒性和培育抗逆新品种提供理论依据。该文介绍了CBF基因家族的结构特点、表达调控及抗寒作用机制,并展望了其在林木抗逆研究中的应用前景。     

植物的冷调节蛋白及诱导其基因表达的条件

人们发现植物在低温锻炼过程中产生的特异性蛋白——冷调节蛋白(coldregulatedproteins,CORPs)与植物的抗寒力密切相关.冷调节基因是一些诱导型表达基因,它们除受低温诱导外,还能接受诸如外源ABA、水分胁迫等外界条件的诱导.在综述这些诱导冷调节基因表达条件的基础上,提出了在低温寒害来临之前有目的地诱导大田植物冷调节基因表达,合成冷调节蛋白,可提高植物抗寒力的新思路.这在农业生产以及园艺植物南树北移工作中具有重要的现实意义.     

巨桉EgrDREB2A基因结构及表达特性分析

【目的】从低温条件下巨桉mRNA抑制消减杂交(SSH)文库中筛选得到一个受低温诱导的基因Egr DREB2A(Eucgr.G03094),通过对其蛋白序列特征、亚细胞定位特点以及低温、ABA和盐胁迫逆境处理条件下基因表达方式的分析,讨论Egr DREB2A在巨桉非生物逆境响应中所发挥的作用。【方法】使用SMART,Mat Inspector和MEGA等生物信息学软件,分析Egr DREB2A的序列和编码蛋白特征,搜寻启动子上的重要顺式作用元件并构建其蛋白序列进化树;亚细胞定位采用基因枪轰击洋葱表皮的方法;Egr DREB2A组织表达特异性及低温、ABA和盐胁迫条件下的基因表达分析分别采用半定量和定量RT-PCR方法进行;在4℃低温下随处理时间(0.5,2,6,12,24,48 h)的基因数字表达谱基础上,用WGCNA和Cytoscape软件对Egr DREB2A进行基因共表达分析。【结果】Egr DREB2A编码的蛋白序列含有1个典型的AP2结构域,且结构域内部包含YRG和RAYD 2个保守区,属于DREB2类基因,进化树构建结果表明该基因属于DREB2类基因的亚型Ⅰ。Egr DREB2A启动子序列上含有多个与植物逆境响应相关的顺式作用元件。该基因有明显的核定位区,亚细胞定位结果也表明其编码蛋白主要在核中表达。正常条件下,根、茎和叶中Egr DREB2A都有表达,但叶片中表达水平相对较高。在不同低温(0,2,4,6,8℃)下Egr DREB2A被强烈诱导,4℃低温下随处理时间(0.5,2,6,12,24,48 h)的延长,其表达量呈上升趋势,不同时间处理下与Egr DREB2A具有共表达关系的基因多属于参与植物对逆境响应的基因。ABA(100μmol·L-1)对Egr DREB2A的表达表现为先促进而后抑制,而Na Cl(200 mmol·L-1)的处理则表现为先抑制后促进。昼夜节律同样影响Egr DREB2A的表达,光照下基因表达升高,黑暗条件下基因表达降低。【结论】巨桉Egr DREB2A属于DREB2类基因,其表达受低温诱导,同时受ABA、盐和昼夜节律的影响。该基因启动子序列上启动子元件及与其共表达基因大多与植物逆境响应有关。这些结果表明Egr DREB2A可能在巨桉抵抗非生物逆境因子的过程中发挥比较重要的作用。     

杨树类锌指基因ZFL的功能分析

以毛果杨叶片cDNA为模板,采用PCR技术分离出杨树ZFL基因,序列分析结果表明该基因序列开放读码框315 bp,共编码104个氨基酸,推导的蛋白质分子量为11.202 kDa,理论等电点9.83,命名为PtrZFL.对ZFL基因进行实时荧光定量PCR表达分析,结果表明:甘露醇、NaCl、H202、ABA、低温胁迫都能诱导PtrZFL基因的表达,且PtrZFL表达量在ABA处理3h时达到最高,然后随处理时间延长而逐渐降低;低温(4℃)胁迫在6h后能显著诱导PtrZFL基因表达,并随着低温处理能一直保持较高水平的表达.根据毛果杨基因组信息设计引物,获得了PtrZFL基因上游1000bp的启动子序列,序列分析结果表明该启动子包含有多个与胁迫相关的元件,如抗冻、缺水、抗寒、脱落酸响应元件ABRE、MYB和WRKY.GUS活性检测发现,该启动子在转基因拟南芥整株中都有表达,但在根部和成熟叶片中表达较强,其他位置表达微弱.     

在低温下ABA对九里香生理生化的影响

为了探明ABA(脱落酸)对九里香的抗寒效应,对九里香离体枝条采用了不同浓度的ABA溶液处理,并分别置于2℃和0℃的低温下处理,以研究其生理生化变化情况．结果表明:采用0．002 5％-0．005 0％的ABA溶液处理,对九里香离体枝条中的Vc含量、POD (过氧化物酶)活性的降低、可溶性糖的消耗、电导率和MDA(丙二醛)的升高等均具有抑制作用;用较高浓度的ABA(0．010 0％)溶液处理九里香,有的则有相反的变化,说明采用0．002 5％-0．005 0％的ABA溶液处理九里香离体枝条．能提高其抗寒能力．     

诱导植物抗寒基因得到充分表达的条件

综述了诱导植物抗寒基因得到充分表达的各种条件，主要包括温度、当照水分、植物激素、化学药剂、物理方法等外界条件，以及植物的生长期、发育水平等内部因素。同时提出了植物抗寒性研究领域中值得深入探讨的问题。     

植物耐盐机理与耐盐植物选育研究进展*

文中结合对植物耐盐生理基础的讨论，综述了耐盐植物，尤其是耐盐树木的选育研究进展。植物对盐分胁迫的反应和适应是一个复杂的生理过程，既有蛋白质、核酸、碳水化合物等结构和能量物质的代谢，还有酶、激素等生长调节物质的合成与激话。在这一系列的反应过程中包含着离子交换与逆向运输，信号刺激与传递．基因活化与合成，其中渗透调节起着关键作用；在渗透调节过程中，离子如K^ 、Ca^2 等在浓度和时空上的变化对渗透调节的启动，调节速率和调节能力都有着重要作用。此外，脯氨酸、脯氨酸甜菜碱、甘氨酸甜菜碱等作为可配伍溶质在调节渗透势变化、整合盐分离子方面有着无可代替的作用。Ca^2 |作为第二信使的一个重要组成部分．在胁迫信号传递方面的功能正受到日益重视。激素如ABA在渗透调节和胁迫基因的诱导方面发挥了重要作用。胁迫基因目前主要是指渗透调节基因。首先在细菌中被发现，后来在高等植物中也相继分离克隆出来。有许多不同的方法用来选育耐盐植物，包括选、引、育等常规育种方法和基因工程，突变体育种等生物工程方法。它们各有特点．都发挥了各自的作用。对植物耐盐机理有限的认识以及高等植物结构与功能的高度复杂性制约着耐盐植物选育工作的进展。     

外源脱落酸处理对库尔勒香梨抗寒性的影响

为给库尔勒地区香梨安全越冬、稳产和高产的相关研究与栽培实践提供理论依据,以树势健壮、结果正常的库尔勒香梨单株为试验材料,在其生长期(2013年5月6日)第1次对其叶面喷施浓度分别为50、70、90 mg/L的外源激素ABA,之后每隔20天喷施1次,共喷施3次;在其休眠期采集枝条样品,在人工低温胁迫条件下测定其膜透性、丙二醛、脯氨酸、可溶性蛋白质、可溶性糖和淀粉含量等抗寒性指标,并采用隶属函数法对其抗寒性进行了综合评价。结果表明:以50 mg/L的ABA喷施1次的枝条其各生理指标值的平均隶属度0.6,其抗寒级别可划为Ⅱ级;以70 mg/L的ABA喷施1次和2次及以50 mg/L的ABA喷施2次与3次的枝条,其各生理指标值的平均隶属度均大于0.4,这几个处理的枝条均为中抗寒型,其抗寒级别均可划为Ⅲ级;以90 mg/L的ABA喷施1次和2次的枝条,其各生理指标值的平均隶属度均大于0.3,这两个处理的枝条均为低抗寒型,其抗寒级别均可划为Ⅳ级;以70和90 mg/L的ABA喷施3次的平均隶属度均小于0.3,这两种处理的枝条均为不抗寒型,其抗寒级别均可划为Ⅴ级。文中认为,以50 mg/L的ABA叶面喷施1次,库尔勒香梨便能抵抗-24℃的低温。     

国外木本植物抗寒性测定方法综述

文中对目前国外采用的不同抗寒性测定方法的测定原理、测定程序以及优缺点等作了较为详细的介绍。这些测定方法有全株冰冻测试法（组织褐变法）、电解质渗出率法、叶绿素荧光法、热分析法（主要用差热分析法）、电阻抗图谱法以及核磁共振显微镜图谱法和可视＋近红外线光谱法等。并简介了这些测定方法的适用性和抗寒性测定方法的研究展望。     

Assessment of seedling storability of Quercus robur and Pinus sylvestris

Pedunculate oak (Quercus robur L.) and Scots pine (Pinus sylvestris L.) seedlings were lifted on several occasions during autumn 1997 to determine the relationships between storability and frost hardiness. On each lifting date their physiological status was determined by assessment of shoot and root electrolyte leakage and frost hardiness, assessed as freeze-induced electrolyte leakage. Additional seedlings were simultaneously cold-stored for field planting and assessment of preplanting root growth potential in April 1998. First year field performance was determined the following winter. Storability and cold acclimation patterns differed between the two species. Both were negatively affected by early lifting, but oak was less sensitive with respect to survival, and pine attained tolerance to cold storage more rapidly and earlier with respect to growth increment. The correlations between shoot frost hardiness and performance suggest that freeze-induced shoot electrolyte leakage (SEL_diff?20) below a threshold of 5% is a good storability predictor for Scots pine in Denmark. A completely reliable criterion for pedunculate oak could not be established.     

Frost hardening spruce container stock for overwintering in Ontario

Difficulties overwintering container stock in northern Ontario led to the development of the extended greenhouse culture hardening regime for spruce seedlings. Laboratories to measure shoot frost hardiness and evaluate terminal bud development were established to monitor nursery crops being hardened using this regime. Information on frost hardiness and bud development provided by these laboratories has been used by nursery managers to determine readiness of container seedlings for overwintering. Since 1982, over 200 stock lots have been monitored by these operational laboratories. This database can be used to determine the importance of nursery cultural factors and seed source on frost hardening. The database shows large differences between nurseries in approach to hardening seedlings which were reflected in levels of freezing damage, winter desiccation and overwintering success. Rates of frost hardening (i.e., the interval between terminal bud initiation and attainment of a --15°C level of shoot frost hardiness) of crops produced in north central Ontario failed to show significant seed source effects. The rate of frost hardening was faster in crops producing fewer needle primordia in terminal buds.     

Short-day treatment during the growing period limits shoot growth and increases frost hardiness of hybrid aspen plants in the nursery

In Finland, under nursery conditions hybrid aspen may continue their shoot growth until early September. Thus, frost hardening is usually delayed. To solve this problem, we used a three-week period of short-day (SD) treatment between late July and mid-August. During autumn after frost exposure, frost hardiness (FH) was assessed three times with a stem-browning test. The re-sults showed that after SD treatment shoot growth ceased and FH increased when compared to untreated hybrid aspen. Furthermore, the height of SD-treated hybrid aspen varied much less than that of the control plants. We conclude that SD treatment in the nursery during the growing period can be used as a supplementary method for producing well-hardened and uniform hybrid aspen plants.     

我国邓恩桉抗寒性研究进展

邓恩桉具有生长快、材性好、耐寒的特点，成为我国桉树北移种植的优良树种。本文概述了国内有关邓恩桉抗寒性的研究动态，讨论了邓恩桉抗寒性与细胞膜系统、保护酶系统等方面的相关性，参考其他植物抗寒性研究方法与思路，对邓恩桉抗寒性的进一步研究提出了一些设想。     

Within‐population variation in autumn frost hardiness and its relationship to bud‐set and height growth in Picea abies

Artificial freezing tests were performed with two sets of 12 full‐sib families of Picea abies. Each set was selected from a complete diallel cross performed within a natural population. Significant differences in autumn frost hardiness were observed between the two populations, which originate from the same altitude and longitude approximately 60 km apart. Substantial variation in frost hardiness was observed within both populations. Significant pheno‐typic correlations at the individual level were found between freezing injury and the two traits terminal bud‐set and height growth year one. However, no significant relationships were present between freezing injury and bud‐set at the family level, indicating that bud‐set cannot be used to predict autumn frost hardiness for families.     

Timing of cold temperature exposure affects root and shoot frost hardiness of Picea Mariana container seedlings

Seventeen‐week‐old black spruce seedlings were hardened under short daylengths and one of three short day length environments, which were either warm (24/16°C, day/night) throughout a 10 week hardening period (WW), cool (10/5°C) throughout hardening (CC), or warm for three weeks followed by seven weeks of cool temperatures (WC). Greatest root and shoot frost hardiness resulted from the exposure of seedlings to three weeks of warm followed by seven weeks of cool temperatures. Seedlings receiving warm temperatures throughout hardening increased in root and shoot frost hardiness, but to a lesser extent than seedlings exposed to cool temperatures. The frost hardiness of woody roots was generally greater than that of fine roots, but the extent of the difference in frost hardiness depended on the time since bud initiation and on the hardening treatment.     

华山松抗寒性的地理变异

本文报道了1980－1988年间26个产地的华山松实生苗和幼树的冻害情况，看到原产云贵高原亚热带山区的华山松，引种到暖温带气候区种植极不抗寒，基本不能越冬，云贵高原区域内种源的抗寒性也有某种程度的差异，尤以西部（云南保山地区）最弱，东部和北部（云南北部、四川西南部和贵州西部）较强，因而不经试验，不宜把云贵高原华山松种子调入暖温带使用。     

The effect of late summer fertilization on the frost hardening of second-year Scots pine seedlings

In this study the effect of summer fertilization on the initiation of frost hardening of containerized second-year Scots pine (Pinus sylvestris L.) seedlings is studied. During the second growing season three different fertilization programs (water soluble NPK with micronutrients) determined by electrical conductivity of peat water extract (0.2, 0.5 and 1.2 mS cm^-1) were initiated. The growth and nutrient concentrations of needles were monitored during the fertilization period. The frost hardiness of seedlings was assessed on four separate occasions at two week intervals from August 7 to September 18. This assessment was based on artificial freezing tests and visual damage scoring of tissue browning on current-year needles. Clear differences in foliar N, P and K concentrations were observed between the fertilization treatments. Fertilization prolonged the growing period of needles and increased root collar diameter. In all the tests, the highest fertilization level resulted in the highest level of frost hardiness. The difference between the fertilization treatments ranged from 1 °C to 2.2 °C. Frost hardiness increased with an increase in foliar nitrogen concentration and slightly less consistently with increases in foliar phosphorus and potassium concentrations.     

Frost hardiness gradients in shoots and roots of picea mariana seedlings

Frost hardiness of tissues along the length of the stem and the root was investigated in first‐year black spruce (Picea mariana (Mill.) B.S.P.) seedlings. Frost hardiness of 1 cm long stem and root segments was evaluated based on Index of Injury, calculated from post‐freezing electrolyte leakage. Frost hardiness was tested approximately weekly beginning seven weeks after seedlings were transferred from an 18 to a 10 h photoperiod, both at day/night temperatures of 26°C/16°C. Trees were transferred to temperatures of 10°C day and 5°C night at a 10 h photoperiod after a further 18 days. Frost hardiness was greater at the terminal bud and least at the root tips. Although shoots were generally more frost hardy than roots, differences in hardiness along the stem and root axes were gradual, rather than abruptly differing at the shoot‐root interface. All tissues, including root tips, increased in frost hardiness after conditioning for 18 days under short photoperiods (10 h) and warm temperatures (26?C/16°C, day/night). Under cold temperatures (10°C/5°C, day/night) all tissues, excepting the root tips, tolerated — 16°C with little subsequent electrolyte leakage.