遮荫与外源脱落酸喷施对粗枝云杉(Picea asperata Mast.)不同种群抗旱性的影响

本研究通过粗枝云杉不同种群进行的温室半控制试验，采用植物生态学、生理学和生物化学的研究方法，系统地研究了粗枝云杉不同种群抗旱性的生长、形态、生理和生化机理，并结合有关研究进行综合分析，得出主要研究结论如下： 1.粗枝云杉对干旱胁迫的综合反应 粗枝云杉在干旱胁迫下的适应机制为：（1）相对生长速率及植株结构的调整：干旱胁迫下虽然植株相对生长速率显著降低，且有相对较多的生物量向根部分配，但并未发现细根/总根比增加。（2）粗枝云杉对干旱胁迫的光合作用表现为：干旱胁迫显著地降低了控制的理想条件下的气体交换，但干旱胁迫对PSII最大光化学效率（Fv/Fm）没有影响，表明干旱并未影响到光合机构。（3）干旱还影响了很多生理生化过程，包括渗透调解物质（游离脯氨酸）、膜脂过氧化产物、脱落酸（ABA）含量的增加，以及保护酶活性的升高。这些结果证明植物遭受干旱胁迫后发生了一系列的形态、生理和生化响应，这些变化能提高干旱时期植物的存活和生长能力。 2.粗枝云杉不同种群对干旱胁迫反应的种群差异 粗枝云杉三个种群－干旱种群（四川丹巴和甘肃迭部）和湿润种群（四川黑水）对干旱适应不同，这种不同应归因于它们采用的用水策略不同：在水分良好和干旱胁迫条件下，受试种群在相对生长速率和水分利用效率（WUE）方面都表现出显著的种群间差异。与湿润种群相比，干旱种群在两种水分条件下有更高的WUE。粗枝云杉不同种群的碳同位素组分（δ13C）只在干旱胁迫下有显著差异，并且这种差异在水分良好时比干旱胁迫条件下小，说明生理响应和干旱适应性之间的关系受植物内部抗旱机制和外部环境条件（如水分可利用性）或两者互作效应的影响。这些结果说明干旱种群和湿润种群所采用的用水策略不同。干旱种群有更强的抗旱能力，采用的是节水型的用水策略，而湿润种群抗旱能力较弱，采用的是耗水型的用水策略。 3. 遮荫对粗枝云杉不同种群抗旱性影响 干旱胁迫显著降低了全光条件下叶相对含水量（RWC）、相对生长速率、气体交换参数、PSII的有效量子产量（Y），提高了非光化学猝灭效率（qN）、水分利用效率、脯氨酸（PRO）积累、脱落酸（ABA）含量及保护酶活性。然而这种变化在遮荫条件下不明显。我们得出结论适度遮荫降低了干旱对植物的胁迫作用。另一方面，在干旱条件下，与湿润种群相比，干旱种群抗旱性更强，表现在干旱种群净光合速率与单位重量上叶氮含量（Nmass）降低较少。另外，干旱种群表现出更为敏感的气孔导度，更高的热耗散能力（qN）能力、用水效率、ABA积累、保护酶活性，以及更低的总用水量、相对生长速率。这一结果表明这两种群采用不同的生理策略对干旱和遮荫做出反应。许多生长和生理反应差异与这两个种群原产地气候条件相适应。 4. 外源脱落酸(ABA)喷施对粗枝云杉不同种群抗旱性影响 外源ABA喷施在干旱和水分良好条件下均不同程度地提高了根/茎比，表明根和茎对ABA敏感程度不同。实验结果还表明，外源ABA喷施对这两个种群在干旱胁迫期间影响不同。干旱胁迫期间，伴随着ABA喷施，湿润种群净光合速率(A)显著降低，而干旱种群净光合速率变化不明显。另一方面，外源ABA喷施显著提高了干旱条件下干旱种群的单位叶面积重（LMA）、根/茎比、细根/总根（Ft）比、水分利用效率（WUE）、ABA含量, 以及保护酶活性。然而，外源ABA喷施对湿润种群的上述测定指标没有显著影响。这一结果表明干旱种群对外源ABA喷施更为敏感, 反应在更大的气孔导度降低，更高的生物量可塑性，及更高的水分利用效率、ABA含量和保护酶活性。综上所述，我们得出结论，粗枝云杉对外源ABA敏感性因种群的不同而不同。该研究结果可为两个明显不同种群在适应分化方面提供强有力的证据。 Arid or semi-arid land covers more than half of China's land territory. In arid systems, severe shortages of soil water often coincide with periods of high temperatures and high solar radiation, producing multiple stresses on plant performance. Protection from high radiation loads in shaded microenvironments during drought may compensate for a loss of productivity due to reduced irradiance when water is available. Additionally, ABA, a well-known stress-inducible plant hormone, has long been studied as a potential mediator for induction of drought tolerance in plants. Picea asperata Mast., which is one of the most important tree species used for the production of pulp wood and timber, is a prime reforestation species in western China. In this experiment, different population of P. asperata were used as experiment material to study the adaptability to drought stress and population differences in adaptabiliy, and the effects of shade and exogenous abscisic acid (ABA) application on the drought tolerance. Our results cold provide a strong theoretical evidence and scientific direction for the afforestation, and rehabilitation of ecosystem in the arid and semi-arid area, and provide a strong evidence for adaptive differentiation of different populations, and so may be used as criteria for species selection and tree improvement. The results are as follows: 1. A large set of parallel response to drought stress Drought stress caused pronounced inhibition of the growth and increased relatively dry matter allocation into the root; drought stress also caused pronounced inhibition of photosynthesis, while drought showed no effects on the maximal quantum yield of PSII photochemistry (Fv/Fm) in dark-adapted leaves, indicating that drought had no effects on the primary photochemistry of PSII. However, in light-adapted leaves, drought reduced the quantum yield of PSII electron transport (Y) and increased the non-photochemical quenching (qN). Drought also affected many physiological and biochemical processes, including increases in superoxide dismutase (SOD), ascorbate peroxidase (APX) activities, malondialdehyde and ABA content. These results demonstrate that there are a large set of parallel changes in the morphological, physiological and biochemical responses when plants are exposed to drought stress; these changes may enhance the capability of plants to survive and grow during drought periods. 2. Difference in adaptation to drought stress between contrasting populations of Picea asperata There were significant population differences in growth, dry matter allocation and water use efficiency. Compared with the wet climate population (Heishui), the dry climate population (Dan ba and Jiebu) showed higher LMA, fine root/total root ratio and water use efficiency under drought-stressed treatments. The results suggested that there were different water-use strategies between the dry population and the wet population. The dry climate population with higher drought tolerance may employ a conservative water-use strategy, whereas the wet climate population with lower drought tolerance may employ a prodigal water-use strategy. These variations in drought responses may be used as criteria for species selection and tree improvement. 3. The effects of shade on the drought tolerance For both populations tested, drought resulted in lower needle relative water content (RWC), relative growth rate (RGR), gas exchange parameters and effective PSII quantum yield (Y), and higher non-photochemical quenching (qN), water use efficiency (WUE), proline (PRO) and abscisic acid (ABA) accumulation, superoxide dismutase (SOD), ascorbate peroxidase (APX) activities as well as malondialdehyde (MDA) levels and electrolyte leakage in sun plants, whereas these changes were not significant in shade plants. Our study results implied that shade, applied together with drought, ameliorated the detrimental effects of drought. On the other hand, compared with the wet climate population, the dry climate population was more tolerant to drought in the sun treatment, as indicated by less decreases in A and mass-based leaf nitrogen content (Nmass), more responsive stomata, greater capacity for non-radiative dissipation of excitation energy as heat (analysed by qN), and higher WUE，higher level of antioxidant enzyme activities，higher ABA accumulation as well as lower MDA content and electrolyte leakage. Many of the differences in growth and physiological responses reported here are consistent with the climatic differences between the locations of the populations of P. asperata. 4. The effects of exogenous abscisic acid (ABA) application on the drought tolerance For both populations tested, exogenous ABA application increased root/shoot ratio (Rs) under well-watered and drought-stressed conditions, indicating that there was differential sensitivity to ABA in the roots and shoots. However, it appeared that ABA application affected the two P. asperata populations very differently during drought. CO2 assimilation rate (A) was significantly decreased in the wet climate population, but only to a minor extent in the dry climate population following ABA application during soil drying. On the other hand, ABA application significantly decreased stomatal conductance (gs), transpiration rate (E) and malondialdehyde (MDA) content, and significantly increased leaf mass per area (LMA), Rs, fine root/total root ratio (Ft), water use efficiency (WUE), ABA contents, superoxide dismutase (SOD), ascorbate peroxidase (APX) and catalase (CAT) activities under drought condition in the dry climate population, whereas ABA application did not significantly affect these parameters in the wet population plants. The results clearly demonstrated that the dry climate population was more responsive to ABA application than the wet climate population, as indicated by the strong stomata closure and by greater plasticity of LMA and biomass allocation, as well as by higher WUE, ABA content and anti-oxidative capacity to defense against oxidative stress, possibly predominantly by APX. We concluded that sensitivity to exogenous ABA application is population dependent in P. asperata. Our results provide strong evidence for adaptive differentiation between populations of P. asperata.

川西亚高山森林生态系统辐射传输研究

辐射传输研究是贯穿森林生态系统的纽带，太阳辐射为植物的生长发育提供光合能量、适宜的环境温度以及发育信息。一方面，气候变化使到达地面辐射能的质和量发生变化，影响到植被的生长发育，改变森林的结构，而森林结构的变化又会影响林冠内辐射能的分配和质量，这些变化会进一步影响到林下土壤温度，改变森林根系活性以及土壤营养转化的效率；连锁反应的结果有可能会使森林生态系统的生产力发生变化，改变碳素和氮素源库的调节方向，从而反馈影响地球气候系统。另一方面，人类作为生态系统的成员，必然需要森林生态系统为其提供更多的原材料和更好的生态服务功能，如何实现这些目标，就需要人类适度调整干预方式和频度，达到预期的目的。本文在建立适合于川西亚高山森林的叶面积测量技术、光照辐射模型和土壤温度变化模型的基础上，对川西亚高山地带森林生态系统的辐射传输特征进行了分析，并从森林结构的角度探讨了林分内的辐射分布以及对土壤温度的影响。主要成果如下： 1. 提出了一种照相法测量叶面积的方法。通过对摆放在平面上的叶片照相，利用投影变化，把非正射图像转化为正射图像，然后经过计算机图像处理得到每一片叶片的面积、周长、长度、宽度等信息。这种方法可使用户以任意方向和距离拍摄处于平面上的叶片，能同时处理大量的叶片，适于野外离体或活体叶片测量。叶片面积分辨率可调，分辨率可以与常用的激光叶面积仪相近甚至更高，而且叶片图像可以存档查询。 2. 提出一种模拟林内光照变化的模型。利用林冠半球照片，记录视点以上半球内的林冠构件空间分布，作为林冠子模型；天空辐射子模型采用国际照明委员会(CIE)的标准晴天和阴天以及插值模型。该模型能够模拟林下某一位点处的实时光斑变化。 3. 提出一种土壤温度变化模型。把土壤视为具有容量和阻力性质的结构，利用电阻和电容器件构建土壤能量分布模型。外界太阳辐射能经过植被以及其它一些能量分配器后进入土壤，其中有一部分转化为土壤势能，即土壤温度。土壤温度的变化类似于电池的充放电过程。在已知模型参数的情况下，可以从太阳辐射计算土壤温度的变化。在模型参数未知的情况下，通过输入和输出值推算模型的参数，而模型参数中的时间常数与土壤组成和含水量有关，这样就可以知道土壤水分的变化情况。 4. 从王朗亚高山森林典型样地林分结构的测量获得林地三维结构图、树冠形态、叶面积密度等参数，这些参数输入到Brunner (1998)开发的tRAYci 模型中计算出一段时间内林分任意位置处的光照值。与林下辐射计测量值以及半球照片计算结果的比较，该模型基本上能够满足对林分光环境了解的要求。 5. 从川西亚高山森林生产力的角度，探讨了森林生产力研究的方法以及川西地区的研究历史和成果，发现了其中的一些规律和问题，特别是在叶面积测量上，还没有使用标准的叶面积指数定义。综合来看，川西地区针叶林叶面积指数(单位土地面积上植物冠层总叶面积的一半) 应在4-5 之间。降雨丰富的华西雨屏带是川西地区森林生产力最高的地区，而向西北森林生产力逐渐降低。川西地区云冷杉林森林生产力平均约为600 gDM m-2 a-1，但是根据辐射能计算的潜在生产力则达到1800 gDM m-2 a-1。实际与潜在森林生产力的巨大差异说明其它因子对生产力的影响。 6. 王朗亚高山3 个典型森林林分中，白桦林样地(BF) 林下草本以糙野青茅、牛至、紫菀等喜阳性物种为主，林下透光度较高；冷杉林样地(FF) 林下透光度最低，以喜阴性物种水金凤、蟹甲草、囊瓣芹等为主；而云杉林样地(SF)林分林龄最大，林下透光度介于冷杉林和白桦林之间，草本层仍然以喜阴性物种东方草莓、紫花碎米芥、酢浆草等为主。冷杉林和云杉林的灌木层也很丰富，卫矛属、五加属、茶藨子属、忍冬属植物很丰富，而在白桦林则以栒摘要子属、榛子属、鹅耳枥属等植物为主。藓类植物在云杉林中最丰富，并且形成毯状层，其它两个林分则很稀少。3 个样地林分结构与林下光环境有很强的相关性，从光环境特征可以在一定程度上推测林分的结构。各样地单纯从乔木层材积推算的NPP 排列顺序为BF>FF>SF，与林下辐射透射率和林分年龄的顺序相同，暗示辐射对群落演替过程的驱动作用。 7. 用半球照相法测得BF、FF 和SF 3 个样地的有效叶面积指数以SF 样地最高，BF 最低。如果考虑针叶树叶片在小枝上的丛聚分布，利用北方针叶林的数值进行校正，则SF 样地LAI 显著增加(达到89%)，其它样地的LAI 基本不变甚至有所下降。校正后的数值与文献中地面测量的结果较相近，说明在使用半球照相法测量川西亚高山针叶林LAI 时必须加以校正。 8. 在3 个样地中，白桦、岷江冷杉和方枝柏种群为丛聚分布，紫果云杉在FF和SF 样地中基本上为随机分布。3 个物种出现丛聚分布的最短距离约为2m，在最短距离以内则为随机分布。最短距离可能与树冠大小有关，种子传播特征以及对光照的需求状况可能是造成这种分布格局类型的原因。 Radiative transfer plays a key role in forest ecosystems. Solar radiation providesenergy for photosynthesis, appropriate ambient temperature and development informationfor plants. However, quality and quantity of radiation reaching land surface are affected byweather and subsequently influence the growth and development of plants, which in turnchanges the budget of radiation in forest. Soil temperature changes with the variation ofradiation under forest canopy and influences the activity of roots and rate of nutrientturnover. Thus, any changes of radiation will induce chain reactions in the entireecosystem and display in the value of net primary productivity which will possibly shiftthe relationship between carbon source and sink at local or regional scale and feed back tothe global climate system. On the other hand, as a component of ecosystems, humanbeings of course need to demand more materials and better service from ecosystems. Forthese purpose, man must adapt their pattern and frequency of interference to ecosystems.This paper aims to research on the canopy structure, the radiation distribution and theirinfluence on soil temperature from the process of radiative transfer in subalpine forestecosystem of western Sichuan. The main results are: 1 Present a new photogrammetric method for leaf area. The main idea is to convertnon-vertically taken images of planar leaves to orthoimages through projectivetransformation. The resultant images are used to get leaf morphological parametersthrough image processing. This method enables users to take photos at almost anyorientation and distance if only the leaves are placed on same plane, and to processlarge quantity of leaves in a short time, which is suitable for field measurement. Theresolution of leaf area is adjustable to fit for special requirement. 2 A model using hemispherical photos combining with solar tracks and radiation courseis provided to simulate light variation in forest. The hemispherical photos of canopyrecord the real spatial distribution of each element of plants viewed from a point. Skyradiance is simulated with CIE standard clear sky or cloudy sky model. This modelcan be used to simulate real time light variation under canopy. 3 Present a soil temperature model. Soil could be regarded as a body of resistor andcapacitor. Some of the budget of solar radiation in soil body is transformed into soilpotential energy, the soil temperature. Variation of soil temperature is driven by solarradiation, vegetation, soil properties, etc. This model has two parameters, one of whichis time constant and is related to soil water content. The inversed model can be used tosimulate the variation of soil water. 4 By using model tRAYci developed by Brunner (1998), the 3-D distribution of light inthree subalpine forest stands of Wanglang Nature Reserve has been simulated andvalidated with value of radiometers in these stands. This model can basically satisfythe need for understanding light regimes of these stands. 5 Present some principles and questions of NPP (net primary of productivity) researchesin western Sichuan. The standard leaf area index (LAI) defined by Chen and Black(1997) has not been used in this region. Total leaf area and projected leaf area indexare still used in NPP researches which may differ around 1-fold in magnitude. Thestandard LAI which is a half of total leaf area above unit land area should be between4 and 5 for typical subalpine coniferous forest of western Sichuan concluded fromliteratures. The maximum forest NPP occurs in West China rain belt and decreasesnorthwestwards. Average NPP of spruce-fir forest in western Sichuan is about600gDM m-2 a-1, which is below the potential NPP of 1800gDM m-2 a-1 based onmeasured radiation in this region. The significant difference between potential and realNPP suggests that other factors influence the growth of stands. 6 In the three subalpine forest stands of Wanglang Nature Reserve, herbage layer ofAbstractbirch stand (BF) with age of 40 is dominated by heliophytes of Deyeuxia scabrescens,Origanum vulgare, Aster tongoloa etc.. However, both of the other two stands aredominated by shade tolerent species, such as Impatiens noli-tangere, Impatiensdicentra, Cacalia deltophylla and Pternopetalum tanakae etc. in fir stand (FF) withage of 180 and Fragaria orientalis, Cardamine tangutorum and Oxalis corniculata etc.in spruce stand (SF) with age of 330. Shrub species in the latter two stands arerelatively rich, typical dominant genera being Euonymus, Acanthopanax, Ribes andLonicera. Birch stand has relatively sparse shrubs dominated by genera of Cotoneaster,Corylus and Carpinus. Mosses are significant only in spruce stand. The canopystructure controls the light regime of stand, which influence the composition of herblayers beneath the canopy. This light regime-community structure relationship can beused to infer the herb community from canopy structure. The NPP derived from timbervolume of arbor layer of the three stands decreases from BF to SF, which is in thesame order of transmitted total radiation under canopy and age of these stands,suggesting the driving effect of radiation in the succession of community. 7 The highest effective LAI of the three stands obtained by hemispherical photos is inplot SF and lowest in plot BF. After rectification of the clumping effect of leaves onshoot, the real LAI in plot SF increases significantly (89%) and approximate to theaverage LAI of coniferous forest in western Sichuan. Therefore, the LAI obtainedfrom hemispherical photos needs rectification for clumping effect. 8 Spatial distribution pattern for Betula platyphylla, Abies faxoniana and Sabinasaltuaria is clumpy, but Picea purpurea almost random in plot FF and SF. The shortestdistance for clumpy distribution for Betula platyphylla and Sabina saltuaria is 1.5m,and 2m for Abies faxoniana. And random pattern for these trees is exhibited within thisrange which almost coincides with the diameter of crown. Seed dispersalcharacteristics and light requirement may be the reason for different spatial pattern.

青杨组(Section Tacamahaca Spach)杨树不同种群对土壤干旱和重金属锰污染的生态生理响应

土壤是人类赖以生存的自然环境和农业生产的重要资源，世界面临的粮食、资源和环境问题与土壤密切相关，目前危害土壤的主要因素是干旱和重金属污染。杨树具有适应性强、生长快和丰产等特性，本论文以青杨组杨树为模式植物，采用植物生态、生理及生物化学等方法，研究杨树对土壤干旱和锰胁迫的生态生理反应以及种群间差异，研究成果可为我国干旱半干旱地区营造人工林、防止沙漠化提供理论依据，也为恢复与重建重金属污染地区退化生态系统提供科学指导。主要研究结果如下： 1. 青海杨不同种群对干旱胁迫的响应差异 干旱胁迫显著降低了两个青海杨种群（干旱种群和湿润种群）生物量积累，包括株高、基径、干物质积累等，通过植物结构的调整，有更多的生物量向根部分配。干旱胁迫还显著降低了叶绿素和类胡萝卜素含量，增加了游离脯氨酸和总氨基酸含量。另一方面，干旱胁迫诱导了活性氧的积累，作为第二信使，激活了抗氧化系统，包括抗坏血酸（ASA）含量和酶系统如超氧化物歧化酶（SOD），愈创木酚过氧化物酶（GPX），抗坏血酸过氧化物酶（APX）和谷胱甘肽还原酶（GR）。这样，杨树既有避旱机制又有耐旱机制，使其在干旱胁迫下有相当程度的可塑性。与湿润种群相比，干旱种群杨树有更多的生物量分配到根部，积累了更多的游离脯氨酸和总氨基酸来进行渗透调节，并且有更有效的抗氧化系统，包括更高含量的ASA 和更高活性的APX 和GR，这些使得干旱种群杨树比湿润种群杨树对干旱有更好的耐性。 2. 喷施硝普钠（SNP）对青海杨阿坝种群干旱胁迫耐性的影响 干旱胁迫显著的降低了青海杨阿坝种群的生长和生物量积累以及叶片相对含水量，还诱导了脯氨酸的合成以进行渗透调节。干旱胁迫下过氧化氢（H2O2）显著累积从而造成对膜脂和蛋白的伤害，使得丙二醛和蛋白羰基含量升高。干旱胁迫下喷施SNP可以减轻干旱胁迫造成的伤害，包括增加叶片的相对含水量，增加脯氨酸和总氨基酸的积累，并激活抗氧化酶系统如SOD，GPX和APX，从而减少丙二醛（MDA）和蛋白羰基（C＝O）的积累，但是在水分良好情况下SNP的效果不显著。 3. 青杨不同种群对锰胁迫的生长与形态响应差异 在同一锰浓度下，干旱种群的耐性指数都要高于湿润种群，这表明青杨对干旱和高锰胁迫具有交叉耐性。两个种群的株高，生物量和叶绿素含量都随锰浓度的升高而逐渐下降。就累积浓度而言，0 和0.1 mM 锰胁迫下，干旱种群积累的锰浓度要高于湿润种群，而在高浓度锰胁迫下(0.5 和1 mM)，湿润种群要高于干旱种群。在0，0.1 和0.5 mM下，锰大多积累在根中，叶片次之，茎中最少。而在1 mM，锰更多的积累在叶片中。就累积总量而言，在各个锰浓度胁迫下，根，茎和叶相比，两个种群青杨都是叶片累积的锰总量要高于根和茎。两个种群间比较，对照中没有显著区别，0.1 mM 锰胁迫下，湿润种群根中累积的锰要高于干旱种群，而在地上部中，干旱种群要高于湿润种群。而0.5 和1 mM 锰胁迫下，根、叶、茎+叶、根+茎+叶中，锰累积总量都是湿润种群高于干旱种群。锰胁迫下，青杨叶片数和叶面积包括总叶面积和平均叶面积都显著降低。叶片横切面的光学显微观察结果表明未经锰胁迫的栅栏组织的细胞饱满，海绵组织发达、清晰；胁迫后杨树叶片栅栏组织细胞出现不同程度的皱缩，海绵组织几乎不可见，此外还发现输导组织在胁迫下密度变小和分生组织严重割裂等现象。 4. 青杨不同种群对锰胁迫的生理与生化响应差异 青杨两个种群脱落酸（ABA）含量在锰胁迫下都显著增加，干旱种群的增幅更大。三种多胺含量在锰胁迫下显示了不同的响应趋势：腐胺在两个种群的各个锰处理下都增加，亚精胺只在干旱种群中显著增加，而精胺除了干旱种群在1 mM 下有所增加外，在锰胁迫下变化很小。谷胱甘肽含量随锰浓度升高而增加，在0.5 mM 锰时达到最高值，1mM 时有所下降。植物络合素（PCs）含量与非蛋白巯基（NP-SH）趋势相似，随锰浓度的升高而增加，且干旱种群中含量要高于湿润种群。锰处理还引起氧化胁迫，表现为过氧化氢和丙二醛含量增加。SOD 活性在湿润种群中，在0 到0.5 mM 锰胁迫下活性升高，但在1 mM 锰胁迫时，其活性有所下降。而在干旱种群中，SOD 活性变化较小，并始终维持在一个较高的水平。APX 活性在两个种群中都随锰浓度的升高而增加，干旱种群活性要高于湿润种群。锰胁迫还显著增加了酚类物质的含量，同时GPX 和多酚氧化酶（PPO）活性也随锰浓度的升高而增加。干旱种群的酚类含量和GPX 与PPO 活性都要高于湿润种群。锰胁迫还改变了氨基酸的含量和构成，根据锰胁迫下浓度变化的不同，可以将游离氨基酸分为三组：第一组包括，谷氨酸，丙氨酸和天门冬氨酸，这一组氨基酸含量在锰胁迫下有所下降。第二组包括缬氨酸，亮氨酸和苏氨酸含量在锰胁迫下基本不变化或变化很小。剩下的氨基酸为第三组，这组氨基酸含量在锰胁迫下显著增加，而根据增加的幅度又可以将它们分为两个亚组，丝氨酸，酪氨酸，苯丙氨酸，组氨酸和脯氨酸，在1 mM 下的含量是对照的4 倍以上。异亮氨酸，赖氨酸，精氨酸和甘氨酸含量在1 mM 下是对照含量的2 倍以下。同时，同一锰浓度下，干旱种群比湿润种群积累的氨基酸含量要高。 Soil is the indispensable environment for human survival and important resource foragriculture development. Food and environmental problems facing the world are all closelyrelated to soil and nowadays it is threatened by many factors, among which drought stress andheavy metal pollution are the most serious ones. Poplars (Populus spp.) are importantcomponents of ecosystem and suitable as a source of fuel, fiber and lumber due to their fastgrowth. In this study, different populations of Section Tacamahaca spach were used as modelplants to investigate the adaptability to drought stress and manganese toxicity and differencesbetween populations from dry and wet climate regions. Our results can provide theoreticalevidence for the afforestation and prevention of desertification in the arid and semi-arid areas,and also can supply scientific direction for the reconstruction and rehalibitation of ecosystemscontaminated by heavy metals. The results are as follows: 1. Differences in ecophysiological responses to drought stress in two contrastingpopulations of Populus przewalskii Drought stress not only significantly affected dry mass accumulation and allocation, butalso significantly decreased chlorophyll pigment contents and accumulated free proline andtotal amino acids. On the other hand, drought also significantly increased the levels ofabscisic acid and reactive oxygen species, as secondary messengers, to induce the entire set ofantioxidative systems including the increase of reduced ascorbic acid content and the activities of superoxide dismutase, guaiacol peroxidase, ascorbate peroxidase and glutathioneredutase. Thus the combination of drought avoidance and tolerance mechanisms conferredpoplar a high degree of plasticity in response to drought stress. Compared with the wetclimate population, the dry climate population showed lower dry matter accumulation andallocated more biomass to root systems, and accumulated more free proline and total aminoacids for osmotic adjustment. The dry climate population also showed more efficientantioxidant systems with higher content of ascorbic acid and higher activities of ascorbateperoxidase and glutathione redutase than the wet climate population. All these made the dryclimate population superior in adaptation to drought stress than the wet climate population. 2. Effect of exogenous applied SNP on drought tolerance in Populus przewalskii Drought stress significantly increased hydrogen peroxide content and caused oxidativestress to lipids and proteins assessed by the increase in malondialdehyde and total carbonylcontents, respectively. The cuttings of P. przewalskii accumulated proline and other aminoacids for osmotic adjustment to lower water potential, and activated the antioxidant enzymes such as superoxide dismutase, guaiacol peroxidase and ascorbate peroxidase to maintain thebalance of generation and quenching of reactive oxygen species. Moreover, exogenous SNPapplication significantly heightened the growth performance of P. przewalskii cuttings underdrought treatment by promotion of proline accumulation and activation of antioxidant enzymeactivities, while under well-watered treatment the effect of SNP application was very little. 3. Morphological responses to manganese toxicity in the two contrasting populations ofPopulus cathayana High concentration of manganese caused significant decrease in shoot height andbiomass accumulation. The tolerance index of the dry climate population was significantlyhigher than that of the wet climate population, suggesting the superior Mn tolerance in theformer and the existence of cross-tolerance of drought stress and high Mn toxicity. Injuries tothe leaf anatomical features were also found as the reduced thickness in palisade and spongyparenchyma, the decreased density in the conducting tissue and the collapse and split in themeristematic tissue in the central vein. As for the Mn concentrations in the plant tissues, under0, 0.1 and 0.5 mM, most of the Mn accumulated in the roots, then leaves, and stem the least, while under 1 mM, most of the Mn accumulated in the leaves. As far as the total amounts ofMn extraction are concerned, the leaf extracted more Mn than the root and stem in the twopopulations under various Mn concentrations. There is no difference between the twopopulations under control. Under 0.1 mM, the wet climate population extracted higher Mn inthe root than the dry climate population, while in the shoot, the dry climate populationextracted much more Mn. Under 0.5 and 1 mM, the wet climate population translocated moreMn both in the root and the shoot than the dry climate population. 4. Physiological and biochemical responses to manganese toxicity in the two contrastingpopulations of Populus cathayana Mn treatment resulted in oxidative stress indicated by the oxidation to lipids, proteinsand DNA. A regulated network of defence strategies was employed for the chelation,detoxification and tolerance of Mn including the enhanced synthesis of ABA and polyamines,the accumulation of free amino acids, especially His and Pro, and the activation of theenzymes superoxide dismutase and guaiacol peroxidase. Contents of non-protein thiol,reduced glutathione, phytochelatins and phenolics compounds and activities of superoxide dismutase, guaiacol peroxidase and polyphenol oxidase also increased significantly forantioxidant or chelation functions. The wet climate population not only accumulated lessabscisic acid, free amino acids, phytochelatins and phenolics compounds, but also exhibitedlower activities of superoxide dismutase, guaiacol peroxidase and polyphenol oxidase thusresulting in more serious oxidative damage and more curtained growth.

三种豆科灌木对干旱胁迫的响应与适应

干旱胁迫是全球范围内影响植物生存、生长和分布的重要环境因子。岷江上游干旱河谷区，由于生态环境的脆弱性和长期人类活动的干扰和过度利用，导致植被严重退化，水土流失加剧，山地灾害频繁，干旱化和荒漠化趋势明显。这种趋势若不能遏制，将严重阻碍区域社会经济的快速协调发展，并且威胁成都平原地区的发展和长江中下游地区的生态安全。因而开展干旱河谷生态恢复研究成为解决这些问题的关键。水分匮乏是限制干旱河谷生态恢复的关键因子，在全球气候变化的背景下，干旱胁迫在区域尺度上可能会更加严重，并使干旱河谷的生态环境更加恶化。因此，深入研究干旱河谷乡土植物对干旱胁迫的响应和适应机理，具有非常重要的理论和实践意义。 本论文以岷江上游干旱河谷的三种乡土豆科灌木，白刺花（Sophora davidii）、小马鞍羊蹄甲（Bauhinia faberi var. microphylla）和小雀花（Campylotropics polyantha）理论和实践意义。为研究对象，在人工控制条件下设计了4-5个连续性干旱胁迫处理，系统地研究了灌木幼苗的生长、生物量积累和水分利用效率（WUE）、形态结构和生理过程等对干旱胁迫的反应，揭示了幼苗的干旱适应能力及种间差异。主要研究结论如下： 1) 灌木生长和繁殖对干旱胁迫的反应 在干旱胁迫下，幼苗生长速率显著减小，叶片衰老和脱落比率增大，这些变化随着胁迫强度的增加具有累积效应。叶片比茎对干旱胁迫的反应更敏感。在严重干旱胁迫下，幼苗的有性繁殖被限制，但在中等程度干旱胁迫下，幼苗的有性繁殖能力被提高。 2) 灌木生物量积累及其分配和WUE对干旱胁迫的反应 在干旱胁迫下，灌木各器官的生物量都显著减小，但是生物量的分配侧重于地下部分，使得根茎比在干旱条件下增大。幼苗的耗水量（WU）随着干旱胁迫的增加而显著减少。白刺花和小马鞍羊蹄甲WUE在干旱胁迫下降低；小雀花的WUE在中等干旱胁迫下升高。 3) 灌木叶片结构特征对干旱胁迫反应 白刺花叶片具有较为典型的旱生型结构，而小马鞍羊蹄甲和小雀花则为中生型结构。在1至2年的干旱胁迫下，灌木叶片结构组成未发生本质性的改变，主要是细胞大小的变化。在中等和严重干旱胁迫下，叶肉组织厚度明显减小；并且气孔和表皮细胞面积也显著减小，气孔和表皮细胞密度显著增大；叶肉细胞层数、P/S值、表皮厚度等无显著变化。 4) 灌木对干旱胁迫的生理响应 气体交换参数和叶片相对含水量（RWC）在中等干旱胁迫下发生了明显的改变，而叶绿素荧光参数和光合色素含量在严重干旱胁迫下才发生显著变化。在干旱胁迫下，净光合作用速率（Pn）、气孔导度（gs）和RWC呈下降趋势，而叶片温度（Tl）呈增加趋势，蒸腾速率（Tr）的变化不明显。除了日最大Pn减小以外，干旱胁迫对气体交换参数的日变化无显著影响，但是对光合-光响应曲线有显著的影响，使有效光合时间缩短。在严重干旱胁迫下光系统受到损害而代谢减弱，PSⅡ中心的内禀光能转换效率（Fv/Fm）、量子产量（Yield）、光化学淬灭参数（qP）显著降低，而非光化学淬灭参数（NPQ）明显增加。气孔限制和非气孔限制对Pn的影响与干旱胁迫强度有关。在中度胁迫下，气孔限制起主导作用，在严重胁迫下非气孔限制起主导作用，40% FC水分条件可能是灌木由气孔限制向非气孔限制的转折点。 5) 灌木对干旱胁迫的适应能力及其种间差异 三种灌木对干旱胁迫具有较好的适应能力，即使在20% FC，幼苗未因干旱胁迫III而死亡；80% FC适宜于幼苗生长。白刺花生长速率慢，耗水量较少，具有较强的耐旱和耐贫瘠能力，并具有干旱忍受机制，能够在较干旱的环境中定居和生长。小马鞍羊蹄甲和小雀花，生长快，水分消耗量较大，尤其是小雀花，对干旱胁迫的忍受能力较弱，具有干旱回避机制，因而适宜于在较为湿润的生境中生长。综合分析表明，生长速率较慢的物种抗旱能力较强，其更适宜于作为干旱地区植被恢复物种。 Drought is often a key factor limiting plant establishment, growth and distribution inmany regions of the world. The harsh environmental conditions and long-termanthropogenic disturbance had resulted in habitat destruction in the dry valley ofMinjiang river, southwest China. Recently, it tended to be more severe on the vegetationdegradation, soil erosion and water loss, natural disaster, as well as desertification, whichimpact on regional booming economy and harmonious development, and would be verydangerous to the environmental security in the middle and lower reaches of Yangzi River.Therefore, ecological restoration in the dry valley is one of the vital tasks in China. Waterdeficit is known to affect adversely vegetation restoration in this place. Moreover, in thecontext of climate change, an increased frequency of drought stress might occur at aregional scale in the dry valleys of Minjiang River. The selection of appropriate plantingspecies for vegetation restoration in regard to regional conditions is an important issue atpresent and in further. The research on responses of indigenous species to drought stresscould provide insights into the improvement of the vegetation restoration in the dry valleys of Minjiang River. In this paper, the responses of three indigenous leguminous shrubs, Sophora davidii,Bauhinia faberi var. microphylla and Campylotropics polyantha, to various soil watersupplies were studied in order to assess drought tolerance of seedlings, and to compare interspecific differences in seedlings’ responses to drought stress. The results were as follows: 1 Growth and reproduction of shrubs in response to drought stress Seedling growth reduced significantly while leaf senescence accelerated underdrought stress, the cumulative responses to prolonged drought were found. The capacityfor reproduction was limited by severe drought stress, and improved by moderate droughtstress. Leaf responses were more sensitive than shoot to various water supplies. 2 WUE, biomass production and its partitioning of shrubs in response to drought stress Drought stress reduced significantly the total dry mass and their components ofseedlings, and altered more biomass allocation to root system, showing higher R/S ratiounder drought. Water use (WU) and water-use efficiency (WUE) of both S. davidii and B.faberi var. microphylla declined strongly with drought stress. The WU C. polyantha ofalso declined with drought stress, but WUE improved under moderate drought stress. 3 Anatomical characteristics and ultrastructures of leaves in response to drought stress There were xeromorphic for S. davidii leaves and mesomorphic for B. faberi var.microphylla and C. polyantha at the all water supplies. The foundational changes in leafstructures were not found with drought stress. However, mesophyll thickness, the areas ofstomatal and epidermis reduced slightly while the densities of stomatal and epidermisincreased under severe drought stress. Variations in these parameters could mainly be duoto cell size. Other structures did not displayed significant changes with drought stress. 4 Physiological responses of shrubs to drought stress The gas exchange parameters and leaf relative water content (RWC) were affectedby moderate stress, while chlorophyll fluorescence and chlorophyll content were onlyaffected by severe stress. Drought stress decreased net photosynthesis rate (Pn), stomatalconductance, light-use efficiency and RWC, and increased leaf temperature. Therespiration rates (Tr) were kept within a narrower range than Pn, resulting in aprogressively increased instantaneous water use effiecency (WUEi) under drought stress.Moreover, drought stress also affected the response curve of Pn to RAR, there was adepression light saturation point (Lsat) and maximum Pn (Pnmax) for moderate andsevere stressed seedling. However, diurnal changes of gas exchange parameters did notdiffer among water supplies although maximum daily Pn declined under severe stress.VISevere stress reduced Fv/Fm, Yield and qP while increased NPQ and chlorophyll content.Photosynthetic activity decreased during drought stress period due to stomatal andnon-stomatal limitations. The relative contribution of these limitations was associatedwith the severity of stress. The limitation to Pn was caused mainly by stomatal limitationunder moderate drought stress, and by the predominance of non-stomatal limitation undersevere stress. In this case, 40% FC water supply may be a non-stomatal limitation 5 Interspecific differences in drought tolerance of shrubs Three shrubs exhibited good performance throughout the experiment process, evenif at 20% FC treatment there were no any seedlings died, 80% FC water supply wassuitable for their establishment and growth. S. davidii minimized their water loss byreducing total leaf area and growth rate, as well as maintained higher RWC and Pncompared to the other two species under drought stress, thus they might be more tolerantto the drought stress than the other two species. On the contrary, it was found that C.polyantha and B. faberi var. microphylla had higher water loss because of their stomatalconductance and higher leaf area ratios. They reduced water loss with shedding theirleaves and changing leaf orientation under drought stress. Based on their responses, thestudied species could be categorized into two: (1) S. davidii with a tolerance mechanismin response to drought stress; (2) C. polyantha and B. faberi var. microphylla withdrought avoidance mechanism. These results indicated that slow-growing shrub speciesare better adapted to drought stress than intermediate or fast-growing species in present orpredicted drought conditions. Therefore, selecting rapid-growing species might leavethese seedlings relatively at a risk of extreme drought.

化感物质香豆素对苜蓿种子萌发及幼苗初级氮同化的影响

香豆素类物质是苯丙酸内酯（环酯）类化合物，绝大部分高等植物通过次生代谢途径都能合成。研究表明，香豆素类物质是花椒体内最重要的化感物质，系统研究香豆素类物质的作用机理有助于理解和最终解决花椒连作障碍。本文通过研究香豆素对几种植物种子特别是苜蓿种子萌发、苜蓿幼苗初级氮同化的影响，从生理生化角度揭示香豆素的作用方式，为花椒连作障碍的解决和化感作用机制的深入理解提供依据。主要研究结果如下：1. 研究了香豆素对6 种常见作物种子萌发的影响，并对一组数据采用4个不同的指标进行评价，对生物测定化感作用中存在的问题进行了讨论。结果发现1.0mM的香豆素对采用的6 种作物的种子萌发均表现出一定的化感作用，4 个指标的敏感程度依次为S (发芽速度)＞AS(累积发芽速度)＞CRG(发芽指数)＞GT(最终发芽率)。种子萌发实验是化感作用研究中最重要、应用最广泛的生物测定方法之一，应根据不同的研究目的合理采用指标和实验方法。2. 采用培养皿试纸法进行种子萌发试验，研究了香豆素水溶液在苜蓿种子萌发过程中对其吸水、电导率及抗氧化保护酶活性的影响。结果表明，影响苜蓿种子发芽的香豆素浓度阀值为0.3mM。香豆素在1.0mM 的浓度下降低了苜蓿种子吸水阶段Ⅱ的吸水速度，使其外渗物质增多，电导率增大，并显著抑制了超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)的活性，同时种子体内丙二醛（MDA）的含量显著增大。高浓度香豆素破坏了膜的结构、影响了抗氧化保护酶的活性是香豆素降低苜蓿发芽率的原因之一，也可能是影响花椒-苜蓿间作的关键因素之一。3. 不同浓度（0、25 μM、50 μM、0.1 mM、1.0 mM）化感活性物质香豆素对10 日龄苜蓿幼苗初级氮同化的影响的结果表明25 µM～50 µM 的香豆素加快了苜蓿幼苗对硝态氮的吸收。高浓度的香豆素导致苜蓿根系和叶片内可溶性蛋白含量降低、鲜重减小、地下鲜重/地上鲜重（R/S）的比值升高，根系中初级氮同化的关键酶硝酸还原酶（NR）、谷氨酸胺合成酶（GS）、谷氨酸脱氢酶（GDH）的活性降低，叶片中NR、GS 的活性减低、叶绿素含量减少，而GDH 的活性升高。香豆素影响苜蓿幼苗氮代谢和氨同化的关键酶，导致体内养分的缺失是香豆素抑制苜蓿幼苗生长的机理之一。Coumarins are lactones of o-hydroxycinnamic acid, and are allelopathiccompounds that originate in the phenylpropanoid pathway. They are synthesized byalmost all higher plants. According to previous studies, coumarins were mostimportant allelochemicals in Chinese prickly ash. Systematically research of theeffect of coumarin could help to comprehend the continuous cropping impediment.The effects of coumarin on seed germination and primary nitrogen assimilation ofalfalfa were studied. The main results showed that:1. We compared four common germination indices (S, AS, CRG, GT)preciously calculated with the same date. The results showed that, at theconcentration of 1.0 mM, coumarin inhibited seeds germination. Among all indices,the S index was most sensitive, followed by the AS and CRG indices. Andsuggestions on the expression of bioassay results were also provided.2. At concentrations above 0.3 mM, coumarin inhibited seed germination in aconcentration-dependent manner. During seed imbibitionⅡ, coumarin at 1.0 mMsignificantly reduced the activities of superoxide dismutase (SOD), catalase (CAT),peroxidase (POD), while the content of malonyldialdehyde (MDA) in alfalfa seedssignificantly increased. The higher concentration coumarin destroyed structure ofmembrane and influenced activities of antioxidant enzymes, which might be one ofthe reasons that coumarin decreased germination rate of alfalfa, and one of the keyfactors influencing Chinese prickly ash-alfalfa intercropping.3. Alfalfa plants were exposed to different concentration of coumarin (0、25μM、50 μM、0.1 mM、1.0 mM) grown for 10 days on control medium. Coumarin, in the range of 25 μM～50 μM, significantly stimulated the net nitrate uptake.Increasing coumarin concentration led to a decrease of protein contents in theleaves and roots. The root to shoot (R/S) FW ratio was increased by increasingcoumarin concentration. Under high coumarin concentration, the activities of nitratereductase (NR) and glutamine synthetase (GS) were repressed in the roots andleaves. Glutamate dehydrogenase (GDH) was inhibited in the roots, while enhancedin the leaves. Chlorophyll contents in the leaves were also decreased under highcoumain concentration. Coumarin decreased alfalfa growth by (i) nutritionaldeficiencies shown by the decrease of nitrate, (ii) lowered N compound synthesisvia inhibition of nitrate reduction and ammonium assimilation.

青藏高原东缘川西云杉林皆伐后灌木生长、繁殖与更新的研究

植物群落及其环境在干扰后的演替格局和过程的研究，是群落和生态系统动态研究的一个热点。选取青藏高原东缘山原区川西云杉林皆伐后，从草地过渡到灌丛的关键阶段的4 个皆伐迹地（恢复时间为8 a、10 a、16 a 和21 a），研究皆伐及自然恢复过程对林下典型灌木银露梅（Potentilla glabra）和唐古特忍冬（Lonicera tangutica）的生长与繁殖能力的影响以及灌木植物在迹地上的更新情况，分析灌木在不同生境中的适应对策和适应能力的差异,为揭示青藏高原东缘山原区迹地植被从草甸到灌丛演替的过程和特点及促进迹地演替与植被恢复进程提供理论依据和技术支撑。研究主要结论如下：1）皆伐后银露梅生长和繁殖能力显著提高，但对唐古特忍冬的影响不明显。皆伐后银露梅丛基径、高度和各部分生物量都显著增加（P < 0.05），但唐古特忍冬只有叶生物量和地下生物量增加，总生物量和其余构件生物量无显著变化。皆伐后，银露梅的结实数量、结实株数、不结实株数和结实株/不结实株比例显著增加（P < 0.05）。自然恢复过程中，银露梅和唐古特忍冬生长能力以及银露梅的结实量都表现出降低的趋势。随着迹地自然恢复时间的增加，银露梅和唐古特忍冬的基径、高度、丛叶片数和各部分生物量有减少的趋势。银露梅的结实株数增加, 但结实数量减少。2）皆伐对银露梅和唐古特忍冬生物量分配模式影响不一致。原始林和迹地中（除CT85）银露梅的生物量大小关系皆为：地下>茎>侧枝>叶。唐古特忍冬在原始林中的生物量大小关系为：茎＞地下部分＞侧枝＞叶,而皆伐后生物量的分配情况改变，生物量大小关系变为：地下部分＞茎＞侧枝＞叶。随着自然恢复时间的增加，银露梅减少了地下生物量的分配，而唐古特忍冬增加了地下生物量的分配。3）皆伐和恢复时间的增加改变了迹地物种组成，促使阳性乔、灌木在迹地上定居。4 个迹地上共出现了灌木15 种，乔木3 种，没有出现天然云杉和冷杉幼苗。随着恢复时间的增加，迹地上的灌木物种由原始林下的耐阴物种逐步发展为以针刺悬钩子（Rubus pungens）为主的阳性灌木。4）影响灌木幼苗密度和幼树密度的因子不一致。灌木幼苗密度与灌木层盖度显著负相关，与苔藓层盖度显著正相关。幼树密度与草本层盖度正相关，与苔藓层盖度、灌木层盖度和高度负相关。5）研究发现在青藏高原东缘山原区皆伐15～20 a 后，迹地仍以草本植物为主，推测皆伐后至少20 a 以上迹地才可能向灌丛阶段过渡，比高山峡谷地区的演替进程至少推迟了20 a。银露梅和唐古特忍冬在皆伐后自然恢复过程中表现出不同的生长与繁殖策略是由两个物种的生物学特性的差异引起的。银露梅比唐古特忍冬更适应迹地退化环境。促进青藏高原东缘山原林区迹地森林恢复一方面是尽量减少人为活动的破坏，另一方面，可以通过在迹地中播种适当的乡土乔、灌木种子（如白桦、银露梅）等人工措施，以加快演替进程。The succession pattern and process of plant community and their environments is a hot spotin community and ecosystem dynamic study. Four clearcuts were chosen in Rangtang(recovery time of 8 a、10 a、16 a and 21 a), which represented the key stage of thecommunity evolved from grass stage to shrub stage in the eastern margin of theQinghai-Tibetan Plateau. The growth and reproduction of the Potentilla glabra andLonicera tangutica and the natural regeneration of shrub plants in the primary Piceabalfouriana forest and 4 clearcuts were studied to explore how clear cutting andnatural recovery process affected the understory shrub species during the 21 years inthe eastern margin of the Qinghai-Tibetan Plateau. The main results were below.1) The growth and reproduction of P. glabra significantly increased after forestclear cutting.. But it was not so significant as to the L. tangutica. The organismbiomass and total biomass of P. glabra were increased obviously after clear cutting（P< 0.05）. But only leaves and underground biomass of L. tangutica increasedsignificantly after clear cutting（P < 0.05）. The number of fruit and growth of P. glabraincreased significantly after clear cutting too（P < 0.05）. The ramet height, basaldiameter , organism biomass and friut number of P. glabra and L. tangutica reducedas the increase of recovery time.2) The biomass allocation patterns varied between P. glabra and L. tangutica inthe primary forest and clearcuts. The biomass allocation of P. glabra both in primary forest and clearcuts was followed as: underground part > stem > branch > leaves.However, the biomass allocation of L. tangutica had changed after the clear cutting.The biomass allocation of L. tangutica in the primary forest was followed as: stem >underground part > branch > leaves and it was underground part > stem > branch >leave in clearcuts. The biomass allocation of P. glabra and L. tangutica varied amongclearcuts. Aboveground biomass was increased while underground biomass decreasedfor P. glabra with the increase of recovery period. However, the L. tangutica showedthe reverse changing pattern.3) Clear cutting and recovery time had changed the species composition of theclearcuts. There were 15.shruby species and 3 tree species in the four clearcuts. Nospruce and fir seedlings were found. In the early stage after clear cutting, there wereonly understory shrub species from the primary Picea balfouriana forest. The sunnyspecies, especieally Rubus pungens invaded intensly as the increase of recovery time.4) There was a significant negative relationship between density of seedlingswith shrub layer coverage and positive correlation with moss coverage. The saplingshad significantly positive correlation with herb layer coverage and negativecorrelation with moss coverage, shrub layer coverage and height.5）Comparing to studies in Miyalou, a nearby high mountain and canyon area,the secondary sucession in this subalpine plateau areas lagged at least 20 years.P. glabra and L. tangutica showed different growth and reproduction strategies toclear cutting and natural recovery , which may associated with the difference of theirbiological characters. P. glabra was more adaptive to the clear cutting than the L.tangutica. Two suggestions were probably recommended to promote the recoveryprogress in the subalpine plateau areas based on the results of this study. Limitanthropogenic disturbance, and meanwhile sow native tree and shrub seeds inclearcuts.

中国西部亚热带常绿阔叶林木本植物小枝功能生态学初步研究

常绿阔叶林以其富饶的生物资源、丰富的生物多样性和巨大的生态与环境效益引起了人们越来越大的重视，它的研究已成为国际植被科学界关注的主题之一。我国分布着世界上面积最大的亚热带常绿阔叶林，在世界植被中具有重要地位，它的分布表现出明显的地带性差异，存在着多样的植物群系及其对应的气候特征。但是在植物功能性状领域，与全球范围其它生物群系相比，常绿阔叶林物种的研究较少，其功能性状间、功能性状与环境间的关系尚不清晰。 本研究以常绿阔叶林木本植物的当年生小枝为对象，试图从小枝水平上的生物量分配格局、叶片大小与数量的权衡关系、小枝茎的构型效应、叶片元素化学计量学，以及小枝大小的成本与效益分析等方面，较为系统地揭示小枝水平上的植物功能性状间及其与气候间的关系。因此，在华西雨屏带内部的不同纬度设置峨眉－青城－雷波－平武的温度梯度进行比较，并对有降水差异的川西南偏湿性（雷波）与偏干性常绿阔叶林（西昌）进行对比研究，同时在不同山体进行不同海拔梯度的比较研究。 本文主要研究结果如下： （1）小枝生物量分配格局叶水平上，叶片重－叶柄重（Y轴vs.X轴,下同）呈斜率小于1的异速生长关系，表明叶柄对叶内部的生物量分配影响显著。小枝水平上，叶和茎的生物量以及它们与小枝总生物量间基本呈等速生长关系，表明大的小枝或大叶物种不一定在叶生物量的分配上占优势。不同生活型间，在小枝或者茎的生物量一定时，常绿物种叶片的生物量比例较落叶物种稍高。与温度和水分较优越（峨眉及其低海拔）的生境相比，在相对低湿（螺髻）与低温（平武）的生境中的植物会减少对叶的投入而增加对支撑部分的投资比例。 （2）小枝叶片大小与数量的权衡无论是不同气候带还是不同生活型以及不同海拔梯度，叶片大小与出叶强度基本都是呈负的等速生长关系，表明了叶片大小－数量在小枝水平上的权衡。在不同气候梯度的对比中，叶片数量（出叶强度）一定时，高温和高水分生境（峨眉）比低温（平武）和低湿（螺髻山）生境中的物种的叶片大小（质量和面积）更大，表明不同生境的比较中，小的叶片可能具有较高的出叶强度和更高的适合度收益。“出叶强度优势”（Leafingintensitypremium）假说可能不适宜解释不同生境物种叶片大小差异。 （3）小枝茎的构型效应虽然茎长和茎径与叶片大小都呈正相关关系，与出叶强度都呈负相关关系，但茎长/茎径比与叶/茎生物量之比呈负相关关系；与叶片的大小呈负相关关系，与出叶强度呈正相关关系。这说明小枝构型能影响小枝叶/茎生物量分配和叶大小－数量的权衡关系。其影响机制可能是小枝内部的顶端优势。另外，茎长/茎径比在低湿和低温等不利生境中的植物中较高，而在降水和温度较适宜环境中较低。 （4）叶片C、N、P化学计量学N含量和P含量，C/N比和比叶重（LMA,leafmassperarea）呈正的等速生长关系，而N和LMA，P和LMA呈负的等速生长关系。在LMA一定时，C/N比随着生境胁迫压力的增加而降低，N、P含量随着生境压力的增加而增加。在P含量一定时，N含量随着生境压力的增加而降低，即N/P比在生境条件较优（峨眉及其低海拔）时较高。常绿和落叶植物叶片的N/P比没有差异，在LMA一定时，常绿植物的N、P含量较高、C/N比较低。总之，植物的C、N、P化学计量学特征受叶片属性如LMA与气候，及其相互作用的影响。 （5）小枝大小的代价与效益分析、TLA与小枝总重总叶面积（TLA,totalleafarea，Y轴，下同）与总叶重（X轴）均呈斜率小于1的异速生长关系，TLA与小枝横切面积呈斜率为1的等速生长关系。表明叶片面积的增加总是小于叶重和小枝总重的增加，随着小枝的增大，它的叶面积支撑效率下降。在热量和降水优越的生境（峨眉及其低海拔）中，相同小枝重或者相同茎横切面积的小枝，其叶面积支撑效率较低湿与低温环境下（螺髻山、平武及高海拔）的高。 总体上，本文初步研究了小枝水平上可能存在的以下三种权衡关系：叶－茎生物量分配权衡；叶片大小－数量的权衡；小枝茎长－茎径的权衡关系，以及气候要素等对这三种权衡关系的影响。在此基础上，我们还讨论了这些权衡关系的可能形成机制，及其与物种生态适应的联系。本研究丰富了生活史对策中关于权衡关系的研究内容，为我国常绿阔叶林功能生态学研究积累了材料。 Evergreen broad-leaved forests are attracting much more attention from vegetation ecologists than ever before because of their abundant nature resource and biological diversity, and also great ecological benefits. China has the largest distribution of subtropical evergreen broad-leaved forests (temperate rainforests) that are typical and representative in the world. The forests span over more than ten degrees in latitude and more than 30 degrees in longitude, providing an ideal place to study plant functional ecology, i.e., the climatic effect on plant functional traits and the relationship between the traits. However, relative to the other biomes, there are few studies addressing functional ecology of the plant species from subtropical evergreen broad-leaved forests. In this study, I focused on the leaf size-twig size spectrum of the woody species of subtropical evergreen broad-leaved forests in southwestern china. I collected data on leaf size and number, twig size in terms of both mass and volume, and stem architecture from five temperate mountains, and then I analyzed the relationships between leaf and stem biomass and between leaf size and number, the effect of stem length/diameter ratio on biomass allocation and on the relationship between leaf size and number, leaf C:N:P stoichiometry, and the twig efficiency of supporting leaf area in relation to twig size. I also addressed the climate effect on the spectrum. The temperature gradient from warm to cool sites was represented by Emei Mountain, Qingchengshan, Leibo, and Pingwu, and the rainfall gradient was assumed to emerge from the comparison between Leibo (High) and Luojishan (Low). In addition, altitudinal effects were analyzed with comparisons between low and high altitudes for each mountains. My main results are as follows. Isometric relationships were found between leaf mass and twig mass and between lamina mass and twig mass, suggesting that the biomass allocation to leaves or laminas was independent of twig mass. Petiole mass disproportionably increase with respect to lamina mass and twig mass, indicating the importance of leaf petioles to the within-twig biomass allocation. In addition, the investigated species tended to have a larger leaf and lamina mass, but a smaller stem mass at a given twig mass at favorable environments including warm and humid sites or at low altitude than unfavorable habitats, which might be due to the large requirements in physical support and transporting safety for the species living at unfavorable conditions. Moreover, the evergreen species invested more in leaves and laminas than the deciduous at given stem or twig biomass within any specified habitats. Negative, isometric scaling relationships between leaf number and size broadly existed in the species regardless of climate, altitude, and life forms, suggesting a leaf size/number trade-off within twigs. Along the climatic gradients, at given leaf number or leafing intensity, the leaves were larger in the favorable environments than the poor habitats. This suggested that the fitness benefit gained by small leaves could be larger than that with high leafing intensity in the stressful sites. I concluded that the “leafing intensity premium” hypothesis was not appropriate to interpreting between-habitat variation in leaf size. Both stem length and diameter were positively correlated to leaf size but negatively correlated to leafing intensity. The ratio of stem length to diameter was negatively correlated to leaf mass fraction, and it was negatively correlated to leaf size but positively correlated to leafing intensity. This suggested that the stem architecture influenced twig biomass allocation and the relationship between leaf size and number. The mechanism underlying the architectural effect might lie in the apical dominance within twig. Moreover, the ratio was greater in unfavorable habitats but smaller in favorable environments. Positive, isometric relationships were found between N and P contents per leaf mass, and between C/N ratio and leaf mass per area (LMA), but N and P contents scaled negatively to LMA. C/N ratio decreased but N and P increased with increasing habitat stress at a given LMA. N content declined with increasing habitat stress at given P content. These indicated that N/P and C/N were higher but LMA was lower in favorable habitats than in the other circumstances. The evergreen and deciduous species were non-heterogeneous in N/P, but the evergreen species have higher N and P contents and lower C/N than the deciduous ones. In general, C:N:P stoichiometry were related to both climatic conditions and other important functional traits like LMA. Total leaf area (TLA) allometricly scaled to leaf mass with a slope shallower than 1, similar to the relationship between TLA and total twig mass (leaf mass plus stem mass), suggesting that TLA failed to keep pace with the increase of leaf mass and twig size. However, TLA scaled isometricly to twig cross-sectional area. Thus, it could be inferred that the twig efficiency of displaying leaf area decreased with increasing twig size. In addition, the efficiency at a given twig size was large in favorable than unfavorable habitats. In general, in this preliminary study, I studied three tradeoff relationships within twigs, i.e., between leaf and stem biomass, between leaf number and size, and between stem length and diameter, as well as the climatic effect on the relationships. I discussed the mechanisms underlying the tradeoff relationships in view of biophysics and eco-physiology of plants. I believe that this study can serve as important materials advancing plant functional ecology of subtropical forest and that it will improve the understanding of life history strategies of plants from this particular biome.

生姜化感作用机理及其栽培模式研究

近年来，随着对作物重茬（连年种植）障碍原因的深入研究，植物的化感作用越来越受到国内外众多学者的重视。而作为重要调料和药用植物的生姜，其连作障碍也备受关注，系统地研究生姜化感作用将有助于理解和最终解决生姜连作障碍问题。本文通过研究生姜不同部位、不同浓度的水浸液对与其间作的两个物种（大豆和四季葱）种子的萌发及幼苗生长的影响，从而证明生姜化感作用的存在；并通过温室盆栽实验研究了生姜的自毒作用（即研究生姜不同部位、不同浓度的水浸液对其幼苗的形态、生理生化、光合作用、土壤酶、土壤微生物多样性及土壤养分的影响），从而揭示生姜退化和衰老的机制，并为生姜筛选出合适的间作物种提供科学依据，对生姜连作障碍提出科学的解决方法。主要研究结果如下： 1. 与对照相比，生姜所有部位（根茎、茎、叶）、所有浓度（10、20、40、 80 g l-1）的水浸液均抑制了大豆种子和葱籽的萌发率、幼苗生长、水分吸收和脂肪酶活性，并且其抑制程度随着水浸液浓度的增加而增强，其生姜各部位水浸液抑制效应的强弱顺序为茎＞叶＞根茎。这一结果表明生姜根茎、茎、叶含有能够抑制大豆种子和葱籽种子萌发和幼苗生长的水溶性化感物质。根茎是生姜的主要收获部位，而生姜的残株（主要是茎和叶）应该从大田中处理掉以减轻其抑制效应。生姜水浸液中主要化感成分包括：根茎水浸液中主要是丁香酸和伞花内脂；茎水浸液中主要是阿魏酸，且其含量最高为73.4 ug/g；叶水浸液中除了阿魏酸，其他六种物质均检测出来，但含量较高的主要有丁香酸、伞花内脂和香豆酸。 2. 生姜茎和叶不同浓度的水浸液均显著抑制了生姜幼苗的株高、每株叶片数和叶面积，其抑制程度随着水浸液浓度的增加而有所增强，而生姜幼苗每株分枝数差异不显著；同时生姜水浸液也极大程度地影响了生姜幼苗的生物量（包括地下生物量、地上生物量和总生物量，均为鲜重）。在同一浓度下，茎水浸液对生姜幼苗形态指标及生物量指标均显示出最强的抑制作用，叶水浸液次之，根茎水浸液最弱。与对照相比，低浓度的生姜根茎水浸液提高了生姜幼苗叶片内四种抗氧化酶（SOD、POD、CAT、APX）活性，高浓度的根茎水浸液抑制了四种抗氧化酶活性，而茎和叶水浸液均随着浓度的增加而抑制了四种抗氧化酶活性，三种水浸液均随着浓度的增加降低了生姜幼苗叶片内叶绿素的含量，而增加了生姜幼苗叶片的相对电导率和丙二醛含量。同时，三种水浸液均随着浓度的增加降低了生姜幼苗的光合参数（包括胞间CO2浓度、气孔导度、蒸腾速率及净光合速率）。 3. 三种生姜水浸液对所测六种土壤酶活性均产生了不同程度的影响，其中影响最大的是酸性磷酸酶和蔗糖酶，在10 g l-1 时就达到了显著水平，并且所有酶均有随着水浸液浓度增加而增大的趋势；相同部位的水浸液随着浓度的增加，细菌和真菌的数量呈增加趋势，而放线菌的数量呈减少趋势；三种生姜水浸液均随着浓度的增加降低了土壤中有机质的含量，加剧了土壤中硝态氮含量的积累，根茎水浸液对土壤有效磷、速效钾和铵态氮均显示出低浓度提高其含量而高浓度降低其含量的趋势，而茎和叶水浸液则随着浓度的增加均降低了其含量。 4. 与生姜单作相比，所有间作系统均在旺盛生长期和收获期不同程度地提高了土壤酶活性，同时也增加了土壤细菌数量及土壤微生物总数但不显著；所有间作系统在旺盛生长期和收获期均不同程度地影响了土壤真菌及放线菌数量（增加或减少），所有间作系统间的多样性指数差异不显著，除了旺盛生长期四种作物（生姜-大豆-四季葱-大蒜）的间作模式显著降低了多样性指数，其值仅为生姜单作的33.18%；生姜与大豆间作不仅提高了19.6%的生姜产量而且获得了较好的经济效益，并且，所有间作系统均显著抑制了生姜姜瘟病的发生。 5. 不同栽培模式不同程度地影响了收获期生姜的株高、分枝数、根茎产量及内在品质。其中处理2显著地促进了生姜的分枝（10.5%），同时处理2、3和4也促进了生姜的生长（株高分别增加了15.0%、11.4%和14.0%），并且这三个处理提高了生姜的产量；处理2和3能有效提高生姜块茎中维生素C（分别较单作生姜显著提高了3.29%和4.05%）、处理3显著提高了可溶性糖（8.2%）、姜辣素（4.6%）和蛋白质等有益物质的含量，降低硝酸盐有害物质的含量（处理2显著降低了14.0%），改善了姜块的外观和内在品质。并且，生姜与大豆间作具有最高的纯收入和产投比，分别较生姜单作提高了24.80%和8.8%。Recently, allelopathy has been more and more paid attentions by national and foreign scholars with profound research on reasons of crop replanted (continuous planted) obstacle. Ginger rhizome is valuable all over the world either as a spice or herbal medicine and ginger replanted obstacle is also paid attentions. Systematic research on ginger allelopathy will contribute to understanding and ultimate solving problem of ginger replanted obstacle. The effects of ginger aqueous extracts with different parts and concentrations on seed germination and early seedling growth of soybean and chive were studied in this article to testify that ginger existed allelopathy. Furthermore, ginger autotoxicity was also studied by pot experiment in greenhouse (namely research on effects of ginger aqueous extracts with different parts and concentrations on morphological indexes, physiological and biochemical indexes, photosynthesis, soil enzymes, soil microbial diversity and soil nutrients) to reveal mechanism of ginger degeneration and senescence, provide scientific basis for selecting appropriate intercropping species and put forward scientific resolvent for ginger replanted obstacle. The main results were as follows: 1. All aqueous extracts at all concentrations inhibited seed germination, seedling growth, water uptake and lipase activity of soybean and chive compared with the control, and the degree of inhibition increased with the incremental extracts concentration. The degree of toxicity of different ginger plant parts can be classified in order of decreasing inhibition as stem＞leaf＞rhizome. The results of this study suggested that rhizome, stem and leaf of ginger contained water soluble allelochemicals which could inhibit seed germination and seedling growth of soybean and chive. The rhizome is the main harvested part of ginger. The residue (mainly stems and leaves) of the ginger plant should be removed from the field so as to diminish its inhibitory effect. The main allelopathic components of three kind of aqueous extracts were as follows: Rhizome extract chiefly contained syringic acid and vmbelliferone and stem extract mainly contained frulic acid whose content was the highest (73.4 ug/g). The other six substances were detected except of frulic acid, but only contents of syringic acid, vmbelliferone and p-coumaric acid were higher. 2. Stem and leaf aqueous extracts of ginger with different concentrations significantly inhibited plant height, leaf numbers per plant and leaf area, and the degree of inhibition increased with the incremental extracts concentration. However, tiller number per plant of ginger seedling showed no significant difference. At the same time, ginger aqueous extracts also influenced biomass including under-ground biomass, above-ground biomass and total biomass (fresh weight) to a large extent. Under the same concentration, stem aqueous extract showed the mostly inhibitory effect on morphological indexes and biomass indexes of ginger seedling. Rhizome aqueous extract showed the leastly inhibitory effect and leaf aqueous extract was intervenient. Enhanced concentration of ginger aqueous extracts significantly reduced total chlorophyll content, accompanying with increases in memberane permeability (REL) and lipid peroxidation (MDA). Compared with the control, rhizome ginger aqueous extract of lower concentration (10 g l-1) increased the activities of major antioxidant enzymes (superoxide dismutase, SOD; peroxidase, POD; catalase, CAT; ascorbate peroxidase, APX) of ginger leaf tissue and higher concentration inhibited the activities of four antioxidant enzymes. However, stem and leaf aqueous extract inhibited the activities of four antioxidant enzymes with increase in concentration. Meanwhile, enhanced concentration of ginger aqueous extracts significantly reduced photo-parameters of ginger seedling (including CO2 concentration, stoma conductivity, net photosynthesis rate and transpiration rate). 3. Rhizome, stem and leaf ginger aqueous extract showed different effect on six soil enzyme activities, and acid phosphatase and invertase showed significant effect when aqueous extract concentration got 10 g l-1. Furthermore, six soil enzyme activities increased with increase in aqueous extract concentration. Bcterial and fungi number tended to increase while antinomyces tented to decrease with the increase in aqueous extract concentration of identical part. Ginger aqueous extracts reduced soil organic matter content with increased concentration, accompanying with NO3-—N accumulation in soil. Rhizome aqueous extract showed the same tendency for available P, available K and NH4+—N, namely lower concentration increased their contents in soil and higher concentration reduced their contents. While stem and leaf aqueous extracts reduced their contents with the increamental concentration. 4. All intercropping systems increased soil enzyme activities to different extent both at VGS and at HS compared to solo ginger. All intercropping systems increased the colony numbers of soil bacteria and total of soil microbe but not significantly either at VGS or at HS. All intercropping systems increased the colony numbers of soil fungi and actinomytes to a different extent (increase or decrease) both at VGS and at HS. For DI, difference between all cultivation patterns and S-G was not significant either at VGS or at HS except that G-S-C-G whose value was only 33.18% of S-G at VGS significantly decreased. G-S not only increased ginger yield by 19.6% but also obtained better economic benefit. Furthermore, all intercropping systems significantly inhibited occurrence of bacterial wilt of ginger. 5. Different cultivated pattern influenced plant height, tiller numbers, rhizome yields and intrinsic quality of ginger. Treatment 2 significantly facilitated tiller occurring (10.5%). Treatment 2, 3 and 4 promoted ginger growth (plant height respectively increased 15.0%、11.4% and 14.0%) and enhanced rhizome yields. Treatment 2 and 3 effectively increased vitamin C content (significantly increased 3.29% and 4.05% compared to solo ginger). Treatment 3 significantly increased contents of beneficial substances such as soluble sugar (8.2%), gingerols (4.6%) and protein. Treatment 2 significantly decreased contents of deleterious substance namely nitrate (14.0%) and improved appearance and intrinsic quality of ginger rhizome. Furthermore, treatment 2 (ginger/soybean intercropping) could obtain better economic benefit and showed the highest net income and ratio of benefit and cost whose values respectively increased by 24.80% and 8.8% compared to solo ginger.

青杨组(Populus Section Tacamahaca Spach)不同种对非生物胁迫的响应差异

土壤是人类赖以生存的自然环境和农业生产的重要资源，目前土壤受到干旱和盐胁迫的危害越来越严重。杨树具有适应性强、生长快和丰产等特性，本论文以青杨组杨树为模式植物，研究杨树对土壤干旱和盐胁迫的生态生理及蛋白质组学反应，研究成果可为我国干旱半干旱地区营造人工林、防止沙漠化提供理论依据，也为恢复与重建盐污染地区退化生态系统提供科学指导。主要研究结果如下： 1 青杨不同种对逐步干旱胁迫的响应差异 将来自喜马拉雅山东缘高海拔的康定杨和低海拔的青杨枝条扦插在温室中，用来检测它们对逐步干旱胁迫的响应。研究结果表明来自不同海拔的杨树对逐步干旱胁迫的适应性反应是不一样的。株高、叶片发育、叶片相对含水量、丙二醛、过氧化氢等指标的显著性变化在青杨中比在康定杨中来得早些，而且随着干旱胁迫程度的增加，这些参数的变化越来越明显，尤其是当青杨受到严重干旱胁迫的时候；而可溶性蛋白、可溶性糖、游离脯氨酸、抗氧化酶活力变化在康定杨中来得早一些。与青杨相比，在干旱胁迫下，康定杨仍能保持较好的植株生长和叶片发育；康定杨也能在逐步干旱条件下积累更多的可溶性蛋白、可溶性糖、游离脯氨酸及抗氧化酶活力，但是在丙二醛和过氧化氢含量方面增加的更少些。而且，我们的研究结果表明高海拔的康定杨有更强的耐干旱能力，杨树对干旱胁迫的适应能力与干旱发生的速度、强度、持续时间及两种杨树的海拔有关。 2 干旱胁迫下青杨不同种的蛋白质组学分析 来自青杨和康定杨雌株的枝条扦插在温室中，用来研究它们对干旱胁迫的蛋白质组学反应。采用TCA-丙酮/酚提取法提取总蛋白，并进行双向电泳分析。在每个处理的重复图像中都能检测到1,000 个以上的蛋白点。在青杨中有58 个蛋白在干旱处理后发生显著变化，其中22 个蛋白通过肽指纹图谱成功鉴定。康定杨中有69 个蛋白的表达量发生了显著变化，其中有25 个蛋白通过肽指纹图谱成功鉴定。这些被鉴定的蛋白主要参与了光合作用、氧化还原平衡、信号传导、能量代谢、蛋白质合成等过程。尽管被鉴定的蛋白只占叶片总蛋白的很少一部分，但这些被鉴定的干旱响应蛋白可能对维持植株内部平衡方面有重要作用。 3 青杨的盐胁迫响应 青杨植株分别用 0、50 和100 mM NaCl 溶液进行处理。叶片相对含水量、叶绿素a、b 含量、CO2 同化速率和气孔导度的降低表明叶绿体受到了盐胁迫的影响。过氧化氢、丙二醛含量及电导率的升高表明细胞受到了伤害。可溶性糖、游离脯氨酸含量及抗氧化酶含量的上升增加了植株耐盐胁迫的能力。在每个处理的重复图像中都能检测到1,000 个以上的蛋白点。其中有38 个盐响应蛋白被成功鉴定，有16 个蛋白(点4、10、11、14、15、21、24、26、27、28、33、34、35、36、37 和38)出现在盐胁迫的植株中；3 个蛋白(点10、11 和35)只出现在重度盐胁迫处理中；而1 个蛋白(点1)只出现在对照处理中。2 个蛋白(点1 和2)表达量下降，其余蛋白点表达量都增加。被鉴定的蛋白一部分参与了生理生化反应，而另一部分则在信号传导、蛋白质合成等方面有重要作用。盐胁迫下的生理生化变化及蛋白质组学的联合研究有利于青杨对盐胁迫的适应性分析。 Soil is the indispensable environment for human survival and important resource for agriculture development. Nowadays soil is threatened by drought stress and salt stress. Poplars (Populus spp.) possess some characters such as strong acclimilation, fast growth and great production of biomass. In this study, different species of Populus section Tacamahaca spach were used as model plants to investigate the ecophysiological and proteomic responses to drought stress and salt stress. Our results can provide theoretical evidence for the afforestation and prevention of desertification in the arid and semi-arid areas, and also can supply scientific direction for the reconstruction and rehalibitation of ecosystems contaminated by salinity. The results are as follows: 1 Adaptive responses to progressive drought stress in two contrasting poplar species originating from different altitudes Cuttings of Populus kangdingensis C. Wang et Tung and Populus cathayana Rehd., originating from high and low altitudes in the eastern Himalaya, respectively, were examined during one growing season in a greenhouse to determine the effects of progressive drought stress. The results manifested that the adaptive responses to progressive drought stress were different in these two species from different altitudes. Significant changes in height increment, leaf development, relative water content (RWC), malondialdehyde (MDA) and hydrogen peroxide (H2O2) appeared earlier in P. cathayana than in P. kangdingensis, whereas changes in soluble protein, soluble sugar, free proline and antioxidant enzymes appeared earlier in P. kangdingensis. In addition, changes in these parameters became more and more significant when the drought stress progressed, especially under severe drought stress in P. cathayana. Compared with P. cathayana, P. kangdingensis was able to maintain a superior height increase and leaf development under drought stress. Also, P. kangdingensis possessed greater increments in soluble protein, soluble sugar, free proline and antioxidant enzymes, but lower increments in MDA and H2O2 than did P. cathayana when the cuttings were exposed to progressive drought stress. Our results suggest that P. kangdingensis originating from the high altitude has a better drought tolerance than does P. cathayana originating from the low altitude. Furthermore, this study manifested that acclimation to drought stress are related the rapidity, severity, duration of the drought event and the altitude of two contrasting species. 2 Proteomic responses to drought stress in two contrasting poplar species originating from different altitudes The cuttings from a female clone of P. kangdingensis and P. cathayana were used to determine proteomic response to drought stress, respectively. Total proteins of the leaves were extracted by a combination of TCA-acetone and phenol, and separated by two-dimensional gel electrophoresis. More than 1,000 protein spots were reproducibly detected on each gel. 58 differentially expressed spots were detected under drought stress in P. cathayana and 22 drought-responsive proteins were identified by peptide mass fingerprint. 69 differentially expressed spots were detected under drought stress in P. kangdingensiss and 25 drought-responsive proteins were identified by peptide mass fingerprint. The identified proteins are involved in several processes, i.e., signal transduction, protein processing, redox homeostasis, CO2 fixation and energy metabolism. Although the proteins identified in this investigation represent only a very small part of the poplar leaf proteins, some of the novel drought-responsive proteins identified here may be involved in the establishment of homeostasis in response to drought stress in the woody plants. 3 Responses to salt stress in P. cathayana Cuttings from a female clone of P. cathayana were treated by Hoagland’s solution: 0, 50, 100 mM NaCl, respectively. Salinity significantly decreased the relative water content of leaves, the contents of chlorophyll a and chlorophyll b, CO2 assimilation rate (A) and stomatal conductance (gs) in both salt stress treatments，which suggested the chloroplast was affected by salt stress. The observed increases of H2O2 and malondialdehyde contents and electrolyte leakage suggested that salinity caused cellular damage, whereas the increases in compatible solutes and in the activities of antioxidant enzymes enhanced the salt tolerance. More than 1,000 protein spots were reproducibly detected on each gel, and 38 salt-responsive proteins were successfully identified by peptide mass fingerprint (PMF). 16 spots (spot 4, 10, 11, 14, 15, 21, 24, 26, 27, 28, 33, 34, 35, 36, 37 and 38) absent in the control sample were induced by the salt treatment, and three spots (spot 10，11 and 35) were present only in the severely salt-stressed treatment. The %vol of the differentially expressed proteins generally increased with progressing salt stress, except for the decreased %vol of two proteins (spot 1 and 2) under salt stress and the presence of spot 1 only in the control sample. Some of the novel salt-responsive proteins identified here may be involved in physiological, biochemical response to salt stress in P. cathayana, the other identified proteins play a role in numerous cellular functions, including signal transduction and protein processing. An integrated physiological, biochemical and proteomic approach was used here to systematically investigate salt acclimation in poplar.

岷江干旱河谷三种蔷薇植物繁殖更新研究

繁殖更新是植物生活史的重要阶段，在退化生态系统中，植物繁殖更新能力往往较差，是植被恢复的限制环节，因而也成为恢复研究重点和核心。本研究选择岷江干旱河谷广泛分布的三种蔷薇：多苞蔷薇（R. multibracteata）、黄蔷薇（R. hugonis）和川滇蔷薇（R. soulieana）为研究对象，通过野外调查，在查明其生长、繁殖更新状况的基础上，采用控制和模拟实验，对种子和幼苗阶段进行了深入研究，综合分析更新潜力，并提出相对应的促进更新和植被恢复措施。主要结论如下： 1）三种蔷薇在岷江干旱河谷广泛分布，生长和繁殖状况良好，结实量大。各生长指标：株高、基径和冠幅，繁殖指标：结实数量、重量和单果重量都具有显著的空间差异性。基径对多苞蔷薇结实量影响最大；而冠幅对黄蔷薇结实量影响最大。海拔和纬度是对蔷薇生长和繁殖影响最大的环境因素，随着海拔和纬度的升高，植株生长更高大，结实量增加；坡度和坡向对其生长和繁殖也有一定影响，随着坡度 和坡向增加，蔷薇生长和结实受到抑制。 2）三种蔷薇在岷江干旱河谷更新现状不佳, 但更新潜力大。活力种子比率低，动物取食以及两年生幼苗的大量死亡是蔷薇更新的主要限制因素。多苞蔷薇和黄蔷薇的结实率低，川滇蔷薇较高。三种蔷薇种子产量大，但种子质量较差，更新具有充足的种源。三种蔷薇都能形成持久种子库，种子库中种子总量大，但有效种子少，黄蔷薇被动物啃食的比例很高，多苞蔷薇和川滇蔷薇也有一部分种子受到动物破坏。三种蔷薇幼苗库组成特征表现为，当年生幼苗所占比例很高，年龄较大幼苗所占比例小。 3）三种蔷薇都具有不同程度休眠，未经处理种子的发芽率极低。黄蔷薇休眠程度最深，为深度生理休眠；多苞蔷薇为中度生理休眠；川滇蔷薇为非深度生理休眠。三种蔷薇种子在形态上发育成熟，种皮具有透水性。蔷薇果果肉和瘦果中含有抑制物质，其浸泡液抑制了油菜种子萌发，果肉抑制作用更强，果肉和瘦果浸泡液的抑制程度分别为：川滇蔷薇>黄蔷薇>多苞蔷薇。切割和硫酸腐蚀提高了川滇蔷薇种子的发芽能力，而对多苞蔷薇和黄蔷薇没有影响。完全去除瘦果果皮和种皮提高了多苞蔷薇种子发芽率，但对黄蔷薇没有影响。赤霉素和烟水对蔷薇种子萌发没有促进作用。三种蔷薇打破休眠所需低温层积时间分别为：黄蔷薇>多苞蔷薇>川滇蔷薇。对于多苞蔷薇和川滇蔷薇，层积前对种子进行硫酸腐蚀或暖温层积能缩短低温层积时间，提高发芽率。对于多苞蔷薇，变温层积中暖温层积和低温层积具有一定的负补性，即延长暖温层积可以缩短种子萌发对低温层积的需要。 4）多苞蔷薇种子形态特征和种子休眠与萌发在不同海拔梯度间存在较大差异。种子采集时间、采集季节和干藏影响多苞蔷薇和川滇蔷薇的种子休眠。多苞蔷薇果实大小、种子大小和千粒重、种皮厚度随海拔升高而增加，而种子饱满率和活力随海拔升高而降低，种子休眠程度也随海拔升高而增加。种皮厚度与种子大小、千粒重成正相关关系，硫酸腐蚀后的种子经过不同时间的低温层积后，种子发芽率与种皮厚度、种子大小、千粒重、海拔成正相关关系。2006 年采集川滇蔷薇和多苞蔷薇种子休眠程度较2005 年低。种子休眠随种子年龄增加而减弱。高温和干旱能减轻多苞蔷薇和川滇蔷薇种子休眠。 5）三种蔷薇的生长和生物量积累在干旱胁迫条件下受到抑制，而生物量分配、叶片形态特征和水分利用特征等都发生了变化。三种蔷薇的根、茎、叶各器官生物量以及总生物量等在干旱胁迫下明显减小，叶片脱落数量增加。在干旱胁迫条件下，较多的生物量分配到地下部分，从而这使R/S 明显增加。比叶面积（SLA）和冠层面积比（LAR）对干旱胁迫的反应不敏感，仅有部分物种在干旱胁迫条件下发生了变化，并且其变化特点在不同年龄幼苗之间有一定差异。干旱胁迫对WUE 的影响在不同物种间存在差异。多苞蔷薇和黄蔷薇的WUE 随着干旱胁迫的增加而增大， 而川滇蔷薇的WUE 则随干旱胁迫增加而减小。在干旱胁迫条件下，多苞蔷薇和黄蔷薇叶片脱落量和生物量减小幅度较川滇蔷薇大，表明其抗旱能力较强。在干旱胁迫条件，三种蔷薇两年生幼苗的生物量减小幅度较当年生幼苗小，表明两年生幼苗的抗旱能力更强。 6）两种植被恢复措施中，幼苗移栽比播种具有更好的植被恢复效果。播种后，蔷薇种子的发芽率较高，但出苗率都很低，即使出苗，幼苗也几乎在一月内全部死亡。 三种微生境条件下（灌木、半灌木和裸地），种子出苗和幼苗成活没有差异。移栽幼苗总体死亡率都比较低，小于20％。特别是两年生幼苗死亡率更低，小于2％。移栽后的幼苗生长状况良好，在整个生长季中，各生长指标不断增加。生境对幼苗的存活率没有显著影响，但对于幼苗的生长和生物量积累有一定影响，裸地更有利于幼苗生长和生物量积累。与当年生幼苗相比，两年生幼苗具有更高的成活率。总之，三种蔷薇在干旱河谷分布广泛、生长繁殖状况良好，结实量大，具有丰富种源，繁殖更新潜力大，但繁殖更新状况不佳；种子散布后动物对种子的取食、种子的深度休眠过程、种子出苗以及当年生幼苗的存活和定居是更新的主要限制环节。水分是影响结实、种子休眠解除和萌发，幼苗存活和定居的最主要的限制因素。在植被恢复中，应在种子成熟季节大量采集种子，在室内打破休眠后进行人工播种，培育两年生幼苗，通过幼苗移栽方式进行植被恢复。川滇蔷薇应栽种在相对湿润的过渡区，而多苞蔷薇和黄蔷薇可以应用于核心区植被恢复。 Regeneration is an important phase in plant life cycle. It has been a key component of ecological restoration in degradation ecosystem in which plants commonly has poor regeneration. In this paper, we investigated the natural growth, propagation and regeneration status of native three rose species, Rosa multibracteata, R. hugonis and R. soulieana, and analysis the limitation in seed germination and seedling establishment stages. Advice on facilitating the use of these plants in restoration based on the results has been proposed. The results were as follows: 1) Three rose plants widely distributed in the dry valley of the Minjiang River, and made a good performance in growth and propagation. There were significant spatial differences in each growth parameter, such as ramet height, basal diameter, crown diameter and propagation parameters including hip number of a clump, hip mass of a clump and a hip mass. Basa diameter was the most important growth parameter influencing fruit number for R. multibracteata and crown diameter was for R. hugoni. Altitude and latitude had the greatest effect on the growth and propagation of rose plants among environmental conditions. Each parameter of growth and propagation increased with the increase of altitude and latitude. In addition, the increase of slope and aspect limited the growth and propagation. 2) Three rose plants had poor natural regeneration, but great regeneration potential. Low seed viability, predation and higher mortality of current year old seedlings were the limitation in regeneration. R. multibracteata and R. hugonis had higher fruiting rates than R. souliean. All three plants produced a great number of seeds, while their viability was poor. Three rose plants had persistent seed banks, with high total seed number but very low viable seed density. Predation was most severe in R. hugonis, and it also existed to some degree in R. multibracteata and R. soulieana. The seedling age-structure was characteristic of current-year seedlings predominating and few older seedlings were observed. 3) Three rose seeds were dormant and untreated seeds germinated with very low germination percentages. The rose seeds had morphological mature embryos, and achenes were permeable. Some inhabit substances existed in hips and achenes for the extracts of hips and achenes inhibited germination of Brassica campestris. The inhibition effect of the extracts of three rose hip and achenes was R. soulieana>R. hugonis>R. multibracteata. Mechanical and H2SO4 scarification increased R. soulieana germination but had no effect on germination of R. hugonis and R. multibracteata seeds. Full removal of pericarp and testa improved the germination of R. multibracteata but did not affect R. hugonis germination. GA3 and smoke water had no positive effect on rose seed germination. The periods of cold stratification required to released seed dormancy was R. hugonis > R. soulieana >R. multibracteata. H2SO4 scarification and warm stratification shortened cold stratification to release dormancy for R. soulieana and R. multibracteata. Warm stratification had complementary effect for cold stratification, i.e. the longer warm stratification seeds received, the shorter cold stratification were required to obtain the same germination percentage. Three rose seeds had different kinds of dormancy; R. hugonis has deep physiological dormancy, R. multibracteata with intermediate physiological dormancy and R. souliean non-deep physiological dormancy. 4）The seeds traits and dormancy of R. multibracteata showed significant difference across altitudes. Year and season of seed collection had significant effect on seed dormancy for both R. souliean and R. multibracteata. Hip size, seed size, seed weight, seed coat thickness and seed dormancy level increased with the increase of the altitude. There were positive relations between seed coat thickness with seed size and seed weight. Germination percentage of seeds treated with H2SO4 scarification following different periods of cold stratification showed positive relation with seed coat thickness, seed size, seed weight and altitude. Seeds of R. souliean and R. multibracteata collected in 2006 had low dormancy level than those collected in 2005. Seed dormancy decreased with increasing seeds age. High temperature and drought were associated with low dormancy level. 5) Seedling growth, the total dry mass and their components of seedlings were reduced, while leaf senescence accelerated under drought stress. More biomass allocation to root system resulted in higher R/S ratio under drought. Water-use efficiency (WUE) of R. multibracteata and R. hugonis increased, while it declined for R. soulieana under drought stress. R. soulieana seedlings had poor drought-resistance capacity it had more senescent leaves, and its reduction of biomass was stronger than two other rose plants under drought. The reduction degree of one year old seedlings under drought stress was slighter than that of current year seedlings. Therefore, one year old seedling was more drought-resistent compared to current year seedlings. 6）Planting seedlings may have better effect in comparison with direct seeding. Most seeds germinated after seeding, but seedling emergence was very low. More than 80 % seedlings from direct seeding died within a months after emergence. Seedling emergence and survival rate did not show difference among microhabitats. Mortality rates of seedlings artificially planted in microhabitats were general lower than 20 %, and the mortality rate of one year old seedlings was lower than 2 %. Each grow parameter including plant height, leaf number and branch number continually increased after planting. Microhabitat type had effect on the growth parameter and biomass production, but it did not influence the seedling survival. Bare land tended to facilitate seedling growth. One year old seedlings had higher survival rate than current year seedlings. In conculsion, the three rose had wide distribution in the dry valley of the Minjiang River. They produced many seeds and had tolerance to drought stress to some degree. But they had poor regeneration in habitats may be caused by predation, seed dormancy，and high mortality in current year seedlings. We recommend that rose plants should be utilized in restoration by planting two-year old seedlings in spring. A large quantity of seeds should be collected artificially in autumn, release seed dormancy in room, and then cultivate two-year old seedlings by seeding in particular container. R. soulieana seedling probably be planted in transition area, and R. multibracteata and R. hugonis can be used in core area of the dry valley of the Minjiang River.

增强UV-B辐射对粗枝云杉(Picea asperata Mast.)和青杨(Populus cathayana Rehd.)的影响

光是植物赖以生存的重要环境因子，但是植物在获得光的同时不可避免的会受到紫外辐射的伤害。尤其是近年来，人类向大气中排放的大量氮氧化合物和氟氯烃类化合物(CFC’s)引起臭氧分子的分解，导致到达地球表面的紫外辐射增加，特别是UV-B辐射增强。而另一方面，植物对UV-B辐射反应的敏感性在种间和品种间存在差异，主要受植物基因型，生态型和生活型的控制。本项目分别以粗枝云杉和青杨组杨树为模式植物，从形态和生理生化方面分别研究了来自不同水分背景下的粗枝云杉种群和来自不同UV-B背景下的青杨种群在增强UV-B下的反应及其反应差异,并探讨了干旱、喷施外源脱落酸(ABA)对它们抗UV-B能力的影响。研究成果可为生态系统的恢复与重建提供理论依据和科学指导。主要研究结果如下： 1. 粗枝云杉的两个种群，湿润种群(来自四川黑水)和干旱种群(来自甘肃迭部)在水分良好和干旱状况下表现出对增强UV-B的不同响应。同时，干旱对粗枝云杉抗UV-B能力的影响也得到研究：两种胁迫共同作用时，干旱表现出在一定程度上减弱了增强UV-B对粗枝云杉的生理特性的影响。 干旱胁迫显著降低了两个粗枝云杉种群的光合同化速率(A), 气孔导度(gs)和PSII的有效光量子产量(Y), 同时，提高了非光化学猝灭效率(qN)和超氧化物歧化酶(SOD)的活性。与湿润种群相比，干旱种群抗旱性更强，表现为干旱种群拥有更高的SOD和干旱进一步加剧了UV-B的胁迫效应。 本研究中，干旱胁迫单独作用时，显著降低了青杨两个种群的生物量积累和气体交换，具体包括A、gs、蒸腾速率(E)和光合氮利用效率(PNUE)，提高了两个种群的瞬时水分利用效率(WUEi)、长期水分利用效率(WUET)、碳同位素组分(δ13C)和氮含量(N)。同时，UV吸收物质和ABA含量也得到积累。另一方面，增强UV-B对青杨两个种群各个指标的影响，同干旱所引起的效应有着相似的趋势。同低海拔种群相比，高海拔种群有着更强的抗旱和抗UV-B能力，具体表现在高海拔种群有着更多的生物量积累，更强的气体交换和水分利用效率及更高水平的ABA和UV吸收物质含量。相比干旱诱导的生物量积累和气体交换的降低，在干旱和增强UV-B两个胁迫同时作用于青杨时，这种降低表现的更为明显。显著的干旱和UV-B的交互作用还表现在WUEi, WUET, δ13C, 可溶性蛋白含量, UV吸收物质含量, ABA, 叶片和茎中的N含量以及C/N比中。 3. 经过一个生长季的试验观察，增强UV-B、外源ABA及两因子共同作用对青杨的生物量积累、气体交换、内源ABA和UV吸收物质含量、抗氧化系统以及碳、氮含量和碳/氮比均产生显著影响。本试验中，青杨的两个种群分别来自中国西南部的不同海拔地区，高海拔种群来自青海大通而低海拔种群来自四川九寨。外源ABA的胁迫为直接喷施ABA到青杨叶片，而增强UV-B胁迫是利用平方波系统分别保证青杨苗暴露于外界UV-B强度和两倍于外界UV-B强度下。 研究结果显示，增强UV-B显著的降低了两个青杨种群的株高、基茎、总叶面积和总生物量等生长指标，同时也导致其A、gs、E和叶片中碳含量的减少。而显著增加了SOD和过氧化物酶(GPx)活性水平，诱导了过氧化氢(H2O2)和MDA的显著增加，促进了UV吸收物质和不同器官中内源ABA含量的显著积累。另一方面，外源ABA引起了青杨光合同化速率的下降，SOD和GPx酶活性的增强，H2O2 和 MDA含量也表现出显著增加，同时，内源ABA含量得到显著累积。同低海拔种群相比，高海拔种群具有更加抗UV-B和外源ABA的特性。显著的UV-B和ABA的交互作用表现在A, E, SOD和GPx活性，以及叶片和根部的内源ABA等一系列指标中。在所有胁迫下，叶片中的碳和氮含量同其在茎和根中的含量显著相关，另外，叶片和茎中的氮含量同茎中的碳含量显著相关。 Sunlight is an indispensable environment factor for plants survival and development. Meanwhile, photosynthetic organisms need sunlight and are thus, inevitably, exposed to UV radiation. Especially for recent years, ultraviolet radiation, especially UV-B reaching the Earth’s surface increased because of depletion of ozone layer resulted from emission of NxO and CFC’s from human activities. On the other hand, the sensitivity of plants to UV-B radiation depends on the species, developmental stage and experimental conditions. In this experiment, two populations of Picea asperata Mast from different water background and two populations of Populus cathayana Rehder from different altitude background were selected as model plants to assess the effects of enhanced UV-B radiation. Morphological and physiological traits induced by enhanced UV-B in each plant species were observed and the different responses were discussed, furthermore the influences of drought and exogenous ABA on responses induced by enhanced UV-B were studied. The study could provide a strong theoretical evidence and scientific direction for the afforestation and rehabilitation of ecosystem. The results are as follows: 1. Different responses of two contrasting Picea asperata Mast. populations to enhanced ultraviolet-B (UV-B) radiation under well-watered and drought conditions were investigated. And the effects of enhanced UV-B on tolerance of drought were also observed in our study that the UV-B exposure may have alleviated some of the damage induced by drought. Two contrasting populations, originating from a wet and dry climate region in China, respectively, were employed in our study. Drought significantly decreased CO2 assimilation rate (A), stomatal conductance (gs) and effective PSII quantum yield (Y), while it significantly increased non-photochemical quenching (qN) and the activity of superoxide dismutase (SOD) in both populations. Compared with the wet climate population, the dry climate population was more acclimated to drought stress and showed much higher activities of SOD and ascorbate peroxidase (APX), and much lower levels of malondialdehyde (MDA) and electrolyte leakage. On the other hand, enhanced UV-B radiation also induced a significant decrease in the chlorophyll (Chl) content in both populations under well-watered conditions, and a significant increase in UV-absorbing compounds in the wet climate population. After one growing season of exposure to different UV-B levels and watering regimes, the increases in MDA and electrolyte leakage, as induced by drought, were less pronounced under the combination of UV-B and drought. In addition, an additive effect of drought and UV-B on A and gs was observed in the wet climate population, and on the activity of APX and qN in the dry climate population. 2. The significant effects of drought, enhanced UV-B radiation and their combination on Populus cathayana Rehd. growth and physiological traits were investigated in two populations, originating from high and low altitudes in south-west China. Our results showed that UV-B acts as an important signal allowing P. cathayana seedlings to respond to drought and that the combination of drought and UV-B may cause synergistically detrimental effects on plant growth in both populations. In both populations, drought significantly decreased biomass accumulation and gas exchange parameters, including A, gs, E and photosynthetic nitrogen use efficiency (PNUE). However, instantaneous water use efficiency (WUEi), transpiration efficiency (WUET), carbon isotope composition (δ13C) and nitrogen (N) content, as well as the accumulation of soluble protein, UV-absorbing compounds and abscisic acid (ABA) were significantly increased by drought. On the other hand, cuttings from both populations, when kept under enhanced UV-B radiation conditions, showed very similar changes in all above-mentioned parameters, as induced by drought. Compared with the low altitude population, the high altitude population was more tolerant to drought and enhanced UV-B, as indicated by the higher level of biomass accumulation, gas exchange, water-use efficiency, ABA concentration and UV-absorbing compounds. After one growing season of exposure to different UV-B levels and watering regimes, the decrease in biomass accumulation and gas exchange, induced by drought, was more pronounced under the combination of UV-B and drought. Significant interactions between drought and UV-B were observed in WUEi, WUET, δ13C, soluble protein, UV-absorbing compounds, ABA and in the leaf and stem N, as well as in the leaf and stem C/N ratio. 3. During one growing season, significant effects induced by enhanced UV-B radiation, exogenous ABA and their combination on biomass accumulation, gas exchange, endogenous ABA and UV-absorbing compounds concentrations, antioxidant system as well as carbon (C) content, nitrogen (N) content and C/N ratio were investigated in two contrasting Populus cathayana populations, originating from high and low altitudes in south-west China. Exogenous ABA was sprayed to the leaves and enhanced UV-B treatment was using a square-wave system to make the seedlings under ambient (1×) or twice ambient (2×) doses of biologically effective UV-B radiation (UV-BBE). Enhanced UV-B radiation significantly decreased height, basal diameter, total leaf area, total biomass, A, gs, E and carbon (C) content in leaves, and significantly increased activities of SOD and guaiacol peroxidase (GPx), hydrogen peroxide (H2O2) and malonaldehyde (MDA) content as well as the accumulation of UV-absorbing compounds and endogenous ABA concentrations among different organs in both populations. In contrast, exogenous ABA showed significant decrease in A and significant increases in activities of SOD and GPx, H2O2, MDA content and the endogenous ABA concentrations. Compared with the low altitude population, the high altitude population was more tolerant to enhanced UV-B and exogenous ABA. Significant interactions between UV-B and ABA were observed in A, E, activities of SOD and GPx, as well as in endogenous ABA in leaves and roots of both populations. Across all treatments, C and N content in leaves was strongly correlated with those were in stems and roots, respectively. Additionally, leaf and stem N content were significant correlated with stem C content.

青杨(Populus cathayana Rehd.)对逆境胁迫的反应及lea基因的表达分析

杨树具有分布广、适应性强，在生态环境治理和解决木材短缺方面均占有重要位置。青杨(Populus cathayana Rehd.)是青杨派树种的重要成员之一，也是我国的特有种。本研究通过对不同水分梯度的干旱胁迫下青杨形态和生理生化的反应，不同pH值盐碱胁迫下不同海拔和不同气候地区的四个青杨种群在生理生态上的反应差异，以及在干旱和低温胁迫下青杨lea2, lea3组基因表达差异的研究，从形态、生理、生化和分子生物学水平系统地研究了青杨在不同逆境胁迫下的反应和青杨不同种群在盐碱胁迫下的反应差异。主要研究结果如下： 1. 青杨在干旱胁迫下的反应机制：中度和重度干旱胁迫下植株的生长受到明显抑制。表现在光合系统上青杨的净光合同化速率(A)下降，主要原因是气孔导度(gs)，胞间二氧化碳浓度(Ci)下降。另外最大量子产量(Fv/Fm)、光化学猝灭效率(qP)降低反应了干旱胁迫下光合系统II(PSII)受到严重损伤, 而且非光化学猝灭效率(qN)上升，导致可利用化学能产量下降，叶绿体产生淀粉的量减少。qP降低qN上升导致产生的过量电子对光合系统的伤害造成活性氧以及丙二醛(MDA)的含量增加。超微解剖结构显示，干旱胁迫增强时，叶绿体内淀粉粒的数目减少，而且叶绿体、线粒体等细胞器中嗜锇颗粒的数目增加。为清除细胞内的活性氧，植物一般的反应是抗氧化系统酶活性增加，对青杨来讲超氧化物歧化酶(SOD), 抗坏血酸过氧化物酶(APx)活性的增加远大于过氧化物酶(POD)，这显示了在青杨中SOD、APx酶在清除活性氧的作用上大于POD。另外同工酶研究结果显示这些酶活性的升高主要是由于各条同工酶带表达量的增加，而不是诱导新酶带的产生。另外，75% FC水分处理下有些指标非但没有下降，像A和有效光量子产量(Y)的值都略有增加，而且gs同时增加。另外，100% FC比75% FC细胞内淀粉粒的数目少一些，但有少量的嗜锇颗粒。这证明100% FC土壤水分也许并非最适合青杨生长。 2. 盐碱胁迫对不同海拔地区青杨种群的反应差异：青杨高海拔和低海拔种群的各种生理特性随着pH值上升都受到了很大的影响。两种群叶和根中Na+、K+ 含量, Na+/K+比率随着pH值的上升影响显著。在pH值高于10.4时高海拔种群叶和根中Na+/K+比率急剧下降但是低海拔种群中却一直维持在较高水平。两种群中MDA、脯氨酸(Proline)的含量，抗氧化系统酶的活性都受到了严重的影响，证明两个种群都属于盐碱胁迫敏感类型但是高海拔的种群对盐碱胁迫的耐性要高于低海拔。这主要是由于高海拔种群一般具有耐干旱、低温胁迫的能力，而植物的抗逆机制一般都有共通之处。 3. 盐碱胁迫对不同气候地区青杨种群的反应差异：盐碱胁迫下两种群的光合作用受到明显的抑制，具体表现在叶绿素的含量和A 显著下降。净光合速率的下降主要是由于叶片gs，Ci 值降低引起的。与湿润地区的种群相比盐碱胁迫增强时，干旱地区的种群叶绿素含量和光合能力的升高与K+离子含量增加有关。植物维持细胞质高K+/Na+值对植物的抗盐性有很重要的作用。为清除盐碱胁迫产生的活性氧，抗氧化系统酶活性增加。盐碱胁迫下干旱地区的种群在SOD、CAT 和谷胱甘肽还原酶(GR)等酶的活性均显著上升，而湿润地区种群只有谷胱甘肽氧化酶(GST)的活性明显增加，说明干旱种群的抗氧化酶系统在较高盐碱胁迫下的保护作用要强于湿润种群。这主要是由于植物抗盐碱胁迫与抗干旱胁迫在一些方面的机制是一致的，抗旱种群一般也能抵抗一定程度的盐碱胁迫。 4. 青杨lea2、lea3 基因在干旱和低温胁迫下的表达差异：通过荧光定量PCR 分析，lea2、lea3 组基因在干旱和低温胁迫下在mRNA 水平的瞬时表达量明显升高，说明了两基因在青杨耐干旱和低温胁迫上都起显著的作用。而且两基因在干旱胁迫下，表达量的升高和降低的时间近乎同步，表明两基因在干旱胁迫下对植物应急保护机制的启动都发挥着重要的作用。低温胁迫下lea3 基因在mRNA 水平上表达量显著上升的时间要早于lea2，而且lea3 基因的持续作用时间明显长于lea2 组基因，说明了低温胁迫开始时lea3基因在植物应对逆境的作用上要大于lea2 基因。 Poplars play an important role in lumber supply, and are important components of ecosystems due to their wide distribution and well adaptation. Populus cathayana Rehd., which belongs to Populus Sect. Tacamahaca Spach, is one of the most important resources of poplars and is specialist to china. In this study, different altitudes and climates populations of P. cathayana were used as experiment materials to investigate the adaptability to drought and salt-alkali stresses. And the cultures of P. cathayana were used to analyze the lea2 and 3 group genes expression when exposed to drought and low temperature stresses. The results are as follows: 1. A large set of parallel responses to drought stress: Drought stress caused pronounced growth inhibition. A decreased significantly and was mainly the result of gs and Ci down. Besides, Fv/Fm, qP decreased and that reflected the harmful effects to PSII of drought stress. In accordance with qN increasing, decreased useful energy production caused the starch numbers reduction in chloroplast. The qP up and qN down improved the levels of ROS and MDA. Starch numbers in chloroplast reduced and plastoglobuli numbers increased when soil water content decreased. To reduce ROS, the activities of SOD, APX, CAT and PPO were activated. The isozymes results show that the rising activities of the antioxidant enzymes resulted from certain isoform content increased, and not from the new band produced. Interestingly, morphological results show 100%FC maybe wasn’t the favorite water content for P. cathayana growth. 2. Effect of salt-alkali stress on morphological and physiological changes in two different altitudes populations of P. cathayana: We compared the physiological responses of two populations of Populus cathayana Rehder, originating from altitudes 2,840 m and 1,450 m. Our results demonstated that Na+ and K+ contents, and Na+/K+ ratios in leaves and roots are greatly affected by pH values. At pH 10.4, the Na+/K+ ratios in both leaves and roots sharply dropped in the higher altitude population but were always maintained at higher levels in the lower altitude population. The pH values causing maximum malondialdehyde (MDA) level, free proline content and antioxidant enzyme activities were significantly different in two populations. These results indicated that the higher altitude population exhibits greater tolerance to alkalinity stress than does the lower altitude population. 3. Morphological and physiological changes in two different climates populations of P. cathayana when exposed to salt-alkali stress. Salt-alkali stress caused pronounced inhibition of the growth and especially in photosystem. Pigments content and A decreased significantly and at the same time gs and Ci decreased too. Compared with wet climate population, the Chlorophyll content and A increased in drought climate population as pH value rising was related to the K+ content increasing. It is important to resist salt-alkali stress that the K+/Na+ ratio matained at high level in cytoplasm. To reduce ROS content, the SOD, CAT and GR activities rised significantly in drought population but only GST increased in wet population. The drought population showed higher salt-alkali tolerance than the wet population mainly resulted from the fact that drought tolerance was in accordance with salt-alkali tolerance to some extent. 4. The different expressional model of lea2 and lea3 gene when P. cathayana was exposed to drought and cold stress. RT-PCR results show both lea2 and lea3 suddenly expressed significantly in mRNA level under drought and cold stress. The expression level of two genes reached optimal level at the same time. But under cold stress, the earlier significantly rising expressional time and the longer maintained higher level time in lea3 than lea2 elucidated that lea3 may be more important than lea2 in resisting cold stress in short time in P. cathayana.

不同抗旱性青稞LEA2/LEA3蛋白基因的克隆与表达

作物的抗旱性是一个多基因控制的、极为复杂的数量性状，植物对干旱在分子水平上的差异反应通过植物组织生理和细胞生物学水平，最终表现为植物抗旱性的不同。在我国，旱地农业超过耕地面积的50%，但水资源短缺，因此培育和选育抗旱高产作物是发展节水型农业最有效的途径。 青藏高原气候恶劣、年均降雨量少，也是世界大麦初生起源中心，因而蕴藏了十分丰富的与抗逆相关的种质资源材料，从这些特殊的资源材料克隆抗旱基因，不仅对培育抗旱、优质、高产大麦新品种具有重要理论意义和经济价值，而且对整个作物抗旱基础和育种应用研究都具重大促进作用。 为了筛选青稞（裸大麦，Hordeum vulgare ssp. vulgare）抗旱性材料，本研究选用来自青藏高原不同地区的84份青稞为材料，在叶片失水率(water loss rate, WLR)检测分析的基础上，选择失水率值差异显著的12个品种，通过相对含水量（relative water content, RWC）和反复干旱法评价其抗旱性，并通过植株对干旱胁迫下的丙二醛（MDA）含量和游离脯氨酸（free-proline）含量变化，了解不同抗旱性材料的生理反应特性。选择抗旱性强弱不同的品种各两份进行LEA2蛋白基因(Dhn6基因)、LEA3蛋白基因(HVA1基因)的克隆，比较LEA蛋白结构差异与作物抗旱性之间的关系。同时，对抗旱性不同的青稞品种受到干旱时间不同的失水变化率（dynamics water loss rate, DWLR）进行了检测；对抗旱性不同的青稞对照材料进行2 h、4 h、8 h和12 h的快速干旱处理，通过SYBR Green实时荧光定量RT-PCR技术对Dhn6基因、Dhn11基因、Dhn13基因和HVA1基因在不同抗旱性材料受到不同干旱时间处理后的相对表达水平进行了检测。本研究对LEA蛋白基因在抗旱性不同的青稞材料中的干旱胁迫分子水平上的差异反应进行了研究，也对植物的抗旱机理进行了初步探讨。主要研究结果如下： 1. 青稞苗期进行离体叶片失水率测定结果表明，来自青藏高原的84份青稞材料的WLR在0.086~0.205gh-1g-1DW之间。选择WLR低于0.1gh-1g-1DW和WLR高于0.18gh-1g-1DW的品种各6份，并对苗期分别进行未干旱及干旱12小时的处理。相对含水量检测结果表明，低失水率青稞材料干旱后的具有更高的相对含水量，盆栽缺水试验也显示叶片失水率低的材料耐旱能力强于失水率高的材料。通过水合茚三酮法测定离体叶片游离脯氨酸的含量，结果表明，所有品种未干旱处理时，游离脯氨酸含量差异不大（17.10~25.74 µgg-1FW）；干旱12小时后，低失水率的品种游离脯氨酸含量明显增高（32.99~53.45µgg-1FW），高失水率品种的游离脯氨酸含量与干旱前变化不明显(P<0.05)。硫代巴比妥酸法测定离体叶片丙二醛(MDA)含量，结果显示，12份所选对照品种中，丙二醛的含量在0.97~2.74nmolg-1FW，干旱12小时后丙二醛的含量显著上升（1.46~4.74nmolg-1FW），高失水率的6个品种的丙二醛含量在未干旱和干旱处理时都明显高于低WLR品种。本研究结果表明青稞的低失水率、低丙二醛含量、高相对含水量和高脯氨酸含量具相关性（P<0.05）。综上研究，我们认为作物失水率的测定可以作为快速检测作物抗旱性的指标之一，因此，强抗旱品种喜玛拉10号(TR1)、品比14号(TR2)和弱抗旱品种冬青8号(TS1)、QB24 (TS2)被选作抗旱基因克隆和表达分析的研究材料。 2. 高等植物胚胎发育晚期丰富蛋白(late embryogenesis abundant proteins, LEA proteins)与植物耐脱水性密切相关，为了探讨青稞LEA蛋白结构差异性与植物抗旱性的关系，本研究以强抗旱品种（喜玛拉10号、品比14号）和弱抗旱品种（冬青8号、QB24）为材料，利用同源克隆法，通过RT-PCR，分别克隆了与抗旱性密切相关的Dhn6基因和HVA1基因。Dhn6基因序列分析结果表明，强抗旱品种品比14号和弱抗旱品种冬青8号Dhn6基因所克隆到的序列为1026bp，它们之间只有5个碱基的差异；喜玛拉10号和QB24克隆到的序列长963bp。在强弱不同的抗旱品种中有22个核苷酸易突变位点，相应的脱水素氨基酸序列推导结果表明，22个核苷酸突变位点中，仅有8个位点导致相应的氨基酸残基的改变，其余的位点系同义突变，另外，21个富含甘氨酸序列的缺失并没有联系作物抗旱性特征。推测这些同义突变位点的氨基酸残基对维持青稞DHN6蛋白的正常结构和功能起着非常重要的作用，也可能DHN6蛋白对青稞长期适应逆境胁迫和遗传进化的结果。对HVA1基因的序列分析结果表明，冬青8号、QB24、品比14号和喜玛拉10号的目的基因核苷酸序列全长分别为661bp、697bp、694bp和691bp，它们都包含1个完整的开放阅读框。相应的LEA3蛋白氨基酸序列结果表明，11个高度保守的氨基酸残基组成基元重复序列的拷贝数与青稞抗旱性之间没有必然关系，在强抗旱品种（喜玛拉10号、品比14号）中三个共同的氨基酸突变位点Gln32、Arg33和Ala195可能对抗旱蛋白的结构和功能有影响；另外，强抗旱青稞品种LEA3蛋白质中11-氨基酸保守基元序列拷贝数和极性氨基酸占蛋白的比例更高，推测LEA3蛋白中基元序列拷贝数和极性氨基酸占蛋白的比例对该蛋白的结构和功能影响更大。 3. LEA蛋白基因的表达水平的上调与植物的耐脱水性密切相关，我们对强抗旱性材料（喜玛拉10号、品比14号）和弱抗旱材料（冬青8号、QB24）进行干旱处理2 h、4 h、6 h、8 h和10 h的失水变化率进行测定，结果表明弱抗旱品种在2~4小时之间失水率变化最明显，而四个对照品种的失水率在8小时后和24小时的失水率值变化不大。进一步提取青稞苗期进行2 h、4 h、8 h和12 h的干旱处理后的总RNA，通过SYBR Green实时荧光定量RT-PCR技术对青稞脱水素基因(Dhn6、Dhn11和Dhn13)和LEA3蛋白基因(HVA1)的相对表达水平受干旱时间和作物抗旱性的影响进行了检测。研究发现，抗旱性不同的青稞品种随干旱处理的时间延长，Dhn6、Dhn11、Dhn13和HVA1基因的相对表达水平不同。 Dhn6基因的相对表达水平在强抗旱青稞品种干旱8小时后快速上升，但在弱抗旱青稞品种干旱处理12小时后检测到更高表达量；Dhn11基因在对照青稞抗旱品种的表达累积水平随干旱时间的延长持续下降；整个干旱过程中，Dhn13基因的相对表达水平在弱抗旱品种持续上升，在强抗旱品种中干旱处理8小时快速上升并达到最高，干旱12小时后降低。与脱水素基因相比较，强抗旱青稞品种在干旱2小时后HVA1基因的相对表达水平显著升高，相对表达量随干旱处理的时间持续上升，在干旱12小时后达到最高；与之相比较，在整个干旱过程中，弱抗旱品种的相对表达水平显著低于强抗旱品种，在干旱8小时之前弱抗旱品种的相对表达水平变化不明显；在干旱8~12小时后却显著上升。上述结果表明，不同的LEA蛋白在植物耐脱水过程中的干旱表达累积水平不同；干旱不是诱导高等植物Dhn11基因表达的主要因素；植物的抗旱性不同，不同LEA蛋白基因对干旱的反应有差异。推测某些LEA蛋白基因的干旱胁迫早期表达累积程度与植物的抗旱性直接相关；其中，Dhn11基因和Dhn12基因不同的表达模式可能与干旱调控表达顺式作用成分(dehydration responsive element, DRE)的有无或结构上的差异有关。 本研究结果认为，(1)失水率和相对含水量可作为植物抗旱性检测的指标之一；(2) DHN6同义突变位点的氨基酸残基对维持该蛋白的正常结构和功能起着重要作用；(3) 11-氨基酸保守基元序列拷贝数和极性氨基酸的比例对LEA3蛋白结构和功能有重要影响；(4)LEA蛋白表达随着干旱胁迫程度而增加，但Dhn11基因并不受干旱诱导表达；(5)作物的抗旱性不同，LEA蛋白对干旱的累积反应并不相同，干旱早期LEA蛋白的累积程度可能会影响植物的抗旱性。 Drought resistance was a complex trait which involved multiple physiological and biochemical mechanisms and regulation of numerous genes. Because its complex traits, it is difficult to understand the mechanisms of drought resistance in plants. Plants respond to water stress through multiple physiological mechanisms at the cellular, tissue, and whole-plant levels. Tibetan hulless barley, a pure line, is a selfing annual plant that has predominantly penetrated into the Qinghai-Tibetan Plateau and remains stable populations there. The wide ecological range of Tibetan hulless barley differs in water availability, temperature, soil type and vegetation, which makes it possess a high potential of adaptive diversity to abiotic stresses. This adaptive genetic diversity indicates that the potential of Tibetan hulless barley serves as a good source for drought resistance alleles for breeding purposes. 12 contrasting drought-tolerant genotypes were selected to measure relative water content (RWC), maldondialdehyde (MDA) and proline content, based on values of water loss rate (WLR) and repeated drought methods from Tibetan populations of cultivated hulless barley. As a result of the screening, sensitive and tolerant genotypes were identified to clarify relationships between characteristics of LEA2/LEA3 genes sequences and expression and drought-tolerant genotypes, associated with resistance to water deficit. In addition, dynamics water loss rate (DWLR) was measured to observe the changes on diffrential drought-tolerant genotypes. Real-time quantitative RT-PCR was applied to detect relative expression levels of Dhn6, Dhn11, Dhn13 and HVA1 genes in sensitive and tolerant genotypes with 2 h, 4 h, 8h and 12 h of dehydration. In the present study, differential sequences and expression of LEA2/LEA3 genes were explored in Tibetan hulless barley, associated with phenotypically diverse drought-tolerant genotypes. 1. The assessments of WLR and RWC were considered as an alternative measure of plant water statues reflecting the metabolic activity in plants, and the parameters of MDA and proline contents were usually consistent with the resistance to water stress. The values of detached leaf WLR of the tested genotypes were highly variable among 84 genotypes, ranging from 0.086 to 0.205 g/h.g DW. The 12 most contrasting genotypes (6 genotypes with the lowest values of WLR and 6 genotypes with the highest values of WLR) were further validated by measuring RWC, MDA and free-proline contents, which were well watered and dehydrated for 12 h. Results of RWC indicated that the values of 12 contrasting genotypes RWC ranged from 89.94% to 93.38% under condition of well water, without significant differences, but 6 genotypes with lower WLR had higher RWC suffered from 12 h dehydration. The results indicated that lower MDA contents, lower scores of WLR and higher proline contents were associated with drought-tolerant genotypes in hulless barley. Remarkably, proline amounts were increased more notable in 6 tolerant genotypes than 6 sensitive genotypes after excised leaves were dehydrated for 12 h, with control to slight changes under condition of well water. Results of MDA contents showed that six 6 tolerant genotypes had lower MDA contents than the 6 sensitive genotypes under both stressed and non-stressed conditions. As a result of that screening, drought- resistant genotypes (Ximala 10 and Pinbi 14) and drought-sensitive genotypes (Dongqing 8 and QB 24) were chosen for comparing the differential characteristics of LEA2/LEA3 genes and their expression analysis. It was conclusion that measurements of WLR could be considered an alternative index as screening of drought-tolerant genotypes in crops. 2. Late embryogenesis abundant (LEA) proteins were thought to protect against water stress in plants. To explore the relationships between configuration of LEA proteins and phenotypically diverse drought-tolerant genotypes, sequences of LEA genes and their deduced proteins were compared in Tibetan hulless barley. Results of comparing Dhn6 gene in Ximala 10 and QB24 indicated that absence of 63bp was found, except that only 5 mutant nucleotides were found. While 22 mutant sites were taken place in Dhn6 gene between sensitive and tolerant lines, 14 synonymous mutation sites appeared in the contrasting genotypes. The additional/absent polypeptide of 21 polar amino acid residues was not consistent with phenotypically drought-tolerant genotypes in hulless barley. It was deduced that synonymous mutation sites would play important roles in holding out right configurations and functions on DHN6 protein. The sequencing analysis results indicated that each cloned HVA1 gene from four selected genotypes contained an entire open reading frame. The whole sequence of HVA1 gene from Dongqing 8, QB24, Pinbi 14 and Ximala 10 was respectively 661bp, 697bp, 694bp and 691bp. Results of DNA sequence analyses showed that the differences in nucleotides of HVA1 gene in sensitive genotypes were not consistent with that of tolerant genotypes, except for absence of 33 nucleotides from +154 to +186 (numbering from ATG) in QB24. Database searches using deduced amino acid sequences showed a high homology in LEA3 proteins in the selected genotypes. Multiple sequence alignments revealed that LEA3 protein from Dongqing 8 was composed of 8 repeats of an 11 amino acid motif, less the fourth motif than Pinbi 14, Ximala 10 and QB24. Consistent mutant amino acid residues appeared in contrasting genotypes by aligning and comparing the coding sequence region, including Gln32, Arg33 and Ala195 in tolerant genotypes as compared to Asp32, Glu33 and Thr195 (Thr184 in Dongqing 8) in sensitive lines. It was concluded that consistent appearance of Gln32, Arg33 and Ala195 would contributed to functions of LEA3 protein in crops, as well as higher proportion of 11-amino-repeating motifs and polar amino acid residues. 3. Most of the LEA genes are up-regulated by dehydration, salinity, or low temperature, are also induced by application of exogenous ABA, which increases in concentration in plants under various stress conditions and acts as a mobile stress signal. Higher levels of proteins of LEA group 3 accumulated was correlated well with high level of desiccation tolerance in severely dehydrated plant seedlings. Dehydrins (DHNs), members of LEA2 protein, are an immunologically distinct protein family, and Dhn genes expression is associated with plant response to dehydration. Dynamic water loss rate was measured between sensitive genotypes and tolerant genotypes after they were dehydrated for 2 h, 4 h, 6h and 8 h. Detailed measurements of WLR at the early stage of dehydration (2, 4, 6, and 8 h) showed that WLR was stabilizing after 8 h, and there were no significant changes between these values and WLR after 24 h. Drought stress was applied to 10-day-old seedlings by draining the solution from the container for defined dehydration periods. Leaf tissues of the selected genotypes were harvested from control plants (time 0); and after 2, 4, 8, and 12 h of dehydration. Differential expression trends of Dhn6, Dhn11, Dhn13 and HVA1 genes were detected in phenotypically diverse drought-tolerant hulless barleys, related to different time of dehydration. Results of quantitative real-time PCR indicated that relative level of HVA1 expression was always higher in tolerant genotypes, rapidly increasing at the earlier stages (after 2-4 h of dehydration). However, HVA1 expressions of sensitive genotypes had a fast increase from 8 h to 12 h of stress. Significant differences in expression trends of dehydrin genes between tolerant genotypes and sensitive lines were detected, mainly in Dhn6 and Dhn13 gene, depending on the duration of the dehydration stress. The relative expression levels of Dhn6 gene were significantly higher in tolerant genotypes after 8 h dehydration, by control with notable higher expression levels after 12 h water stress in sensitive ones. The relative expression levels of Dhn13 gene tended to ascend during exposure to dehydration in drought-sensitive genotypes. However, fluctuate trends of Dhn13 expression level were detected in drought-resistant lines, including in lower expression levels of 12 h dehydration as compared to 8 h water stress. It was conclusion that (1) diverse LEA proteins would play variable roles in resisting water stress in plants; (2) expression of Dhn11 gene was not induced by dehydrated signals because of the trends of expression descended in contrasting genotypes suffered from water deficit and (3) variable accumulations on LEA proteins would be appear in diverse drought-tolerant genotypes during dehydrations. It is deduced that higher accumulations of Dhn6 and Dhn13 expression in 8 h dehydration are related to diverse drought-tolerant lines in crops. The present results indicated that different dehydrin genes would play variable functional roles in resisting water stress when plants were suffered from water deficit. The authors suggest physiologically different reactions between resistant and sensitive genotypes may be the results of differential expression of drought-resistant genes and related signal genes in plants. In addition, contrarily induced expression of Dhn11 and Dhn12 was related to dehydration responsive element (DRE) in barleys. The present study indicated that (1) measurements of WLR and RWC could be considered as one index of drought-tolerant screenings; (2) synonymous mutation sites would play important roles in holding out right configurations and functions on DHN6 protein, (3) higher proportion of 11-amino-repeating motifs and polar amino acid residues would contribute to functions on LEA3 protein, (4) the longer drought, the more accumulation on LEA proteins, except for Dhn11 gene in crops and (5) differential responses on expression of LEA protein genes would result in physiological traits of drought tolerance in plants.

在盐胁迫下外源维生素浸种对小麦发芽及幼苗生长的影响

维生素(Vitamin)又称维他命，为“万年青”产品，是维持人体生命健康必需的一类低分子有机化合物质。维生素对人体健康的作用人们研究很多, 维生素可以增强人体对感染的抵抗力，降低出生缺陷及降低癌症和心脏病发病率等，一旦缺乏，肌体代谢就会失去平衡，免疫力下降，各种疾病，病毒就会趁虚而入；而维生素对作物影响的研究却很少。目前为止，尚无对用维生素浸种的方法来研究外源维生素是否对小麦种子萌发及幼苗生长起调节作用的报道，且对其在小麦抗逆性方面影响的研究甚少，对盐的胁迫抗性研究尚未有人报道。小麦（Triticum aestivum L.）属于拒盐的淡土性作物。盐害不利于小麦生长，严重影响小麦的产量和品质。本研究采用4 种不同维生素VB1、VC、VB6、VPP，分别对供试小麦品种川育12（红皮）、川育16（白皮）小麦浸种后，在一般自然条件下和逆境（盐胁迫条件）下，进行试验。探讨在正常情况下与在不同盐浓度条件下，各维生素及盐浓度对小麦发芽及幼苗生长的影响，并且比较两种不同皮色的小麦在相同盐胁迫条件下的差异表现，同时研究维生素处理的特异性，且哪种维生素对盐害缓解作用最佳。研究结果表明：在无盐胁迫（自然）条件下，对用4 种不同维生素VB1、VC、VB6、VPP 浸种小麦川育12、川育16 后的种子萌发及幼苗生长（幼苗的根长、根重、苗高、苗鲜重）的研究结果表明：4 种外源维生素浸种均对小麦发芽有调节作用，都能提高其最终发芽率。但是提高幅度有所差异。用VB6 浸种后的小麦提高幅度最多，VC 次之，VPP 提高幅度最小。同时，4 种外源维生素浸种对小麦种子的出芽速度及芽后长势也有一定的影响。VB6、VC 处理的小麦种子出芽速度最快，萌发后长势最好；VB1 出芽速度相对较慢，VPP 最慢,但都大于对照；VB1 处理长势略高于对照，VPP 处理的小麦长势则低于对照。从整体来看，VB6、VC处理促进效应明显， VB1 次之，而VPP 在某些方面无效甚至产生负效应。此外，相同的维生素处理对不同的品种的种子萌发、生长效果也存在差异，各种维生素作用于川育12 的效应均强于对川育16。进一步对幼苗根系TTC 还原力及幼苗叶片中硝酸还原酶活性进行测定、分析。研究发现：并非所有种类的维生素对幼苗根系TTC 还原力及幼苗叶片中硝酸还原酶活性的提高都有帮助。幼苗根系TTC 还原力在不同维生素处理下存在显著差异，而与小麦品种关系甚微。经VB6、VC 处理后，根系TTC 还原力测定值均显著高于对照，VB1 不明显，VPP 则略低于对照。VB6、VC 处理的幼苗叶片中硝酸还原酶的含量大于对照，VB1 与对照相差无几，而VPP 处理的川育12 幼苗叶片中的硝酸还原酶活性比对照CK 略高，而在川育16 中则略比对照CK 有所下降，呈现出抑制效应。综上结果表明：VB6、VC 具有促进种子发芽，幼苗生长及根系生长的作用，是较好的促生长剂；VPP 具有抑制作用，是较好的抑制剂，可进一步研究、开发利用。在盐胁迫条件下，对用4 种不同维生素VB1、VC、VB6、VPP 浸种川育12、川育16 后的种子萌发及幼苗生长（幼苗的根长、根重、苗高、苗鲜重）的研究结果表明：在不同盐浓度胁迫条件下， 各处理的种子萌发及幼苗生长均受到不同程度的抑制。随着盐浓度的增加, 发芽率、发芽指数和活力指数成下降趋势；幼苗的根长、根重、苗高、苗鲜重不断降低。4 种维生素处理间也表现出较大差异。VB6、VC 在每个处理中均保持对盐害的缓解作用，VB6 较VC 更易于促进发芽及幼苗生长。最终发芽率高，根系多、长、重，苗高高、重。而VB1、VPP 则表现出抑制作用。在高盐浓度150mM 时，4 种维生素浸种后的种子，其最终发芽率均不能达到40％，但VB6、VC 处理最终发芽率、苗重、根重均高于对照，VPP 最终发芽率、苗重、根重均低于对照。进一步对幼苗根系TTC 还原力及幼苗叶片中脯氨酸含量进行测定、分析。研究发现：不同盐浓度，不同维生素处理、不同品种间存在差异。随着盐浓度的增加（75mM，100mM，150mM），幼苗根系TTC 还原力活性成下降趋势，幼苗叶片中脯氨酸的积累量成上升趋势。VB6 处理脯氨酸含量增加最为明显，VC 次之，VPP 与对照接近，其变化幅度最小。经VB6、VC 处理后的幼苗根系还原强度，在不同盐浓度下，测定值均显著高于对照，VB1 不明显，VPP 则低于对照，产生负效应。此外，品种间表现不尽相同，相同的维生素处理，相同的盐浓度对不同的品种的种子萌发、生长效果也存在差异， 4 种维生素对川育16 的作用均强于川育12，但其影响趋势是一致的。说明VB6、VC 具有耐（抗）盐性，可以促进种子发芽和幼苗生长，是较好的耐（抗）盐拌种剂。 Vitamin is one kind of necessary low molecular compound for humans tosustain health and life. Lots of Studies have been done on the effectc of the vitaminsfor people. Vitamin can help people improve the body's natural resistance to disease,Drop the rate of birth defects、cacers and the incidence of the heart diseases. Ifpeople have less of them, the metabolism of the organism may throw off balance,immunity may drop off, and catch disease; Though the effects for Vitamin to thecrops are limited. up to now, there’s no one use soking seeds of wheats with vitaminsas a method, to study on how the effects will happen on the wheat seed germinationand seedling growth, and there are only few reserches on antireversion force forwheats ,none for the antireversion force in Sault stress condition.Wheat（Triticum aestivum L.）is sensitive to the salt, so the salt damage will doharm to wheat’s growth, it will have an unfavorable impact on the output and thequality of wheat.On this reaserch, we Soaking CHY12(red)、CHY16 (white) wheat seeds withVitamin C, B1, PP, B6 (50mg/L) as a pretreatment first. Then under two condition: one is in the normal environment the other is in different Salinity, we begin ourexperiments. Then disscuss on if the vitamin and salinity affect the wheat seedgermination and seedling growth, and what is the different between the two of them,the result shows that:Under the normal condition, after soaking seeds with VB1、VC、VB6、Vpp，we study on the their seed germination and the seeding growth(the root length andweights, The seedling heights and weights), it shows that all of those four kinds ofvitamin can adjust the seed germination, but different in The growth rate. VB6 isbest for increase, VC comes second,VPP is the worst. Meanwhile, those four vitaminalso have effect on the speed of the sprouting of the wheat. VB6、Vc can faster theseed germination most, and the seedlings are all doing well; VB1 do little effects onthe budding, Vpp is the worst, but all treatments are better than CK; but in Vi, VB1some what above the CK, while VPP lower than that. On the whole, the acceleratingeffect of VB6、VC are obvious, VB1 takes second place, but VPP in some aspects arenoneffective even have negative effect. Furthermore, different kind of seeds with thesame vitamin may different in seed germination and seedling growth, four vitaminson CHY16 is better than CHY12.More studies on TTC reductive capacity of roots and the activity of nitratereductase in the leaves, the reasult shows not all the vitamin can help the seedlings toimprove the TTC reductive capacity and the activity of nitrate reductase. TTCreductive capacity in different treatments shows significant differences,but notcorrelate to the variety of the wheat. The TTC reductive capacity of VB6、Vctreatments are all higher than CK, VB1 is nearly the same as CK, VPP is a littlelower than CK. Through the study of acivity of nitrate reductase, it shows that,VB6、VC are higher than CK ,VB1 is nearly the same as CK also, VPP is a little higher inthe CK of CHY12 but lower in CHY16. Through all the results above: VB6、Vc helpthe wheat seed germination, seedling growth and the growth of roots, is theperfectable factor of stimulating the growth; Vpp is a inhibition, that’ll be furtherreserch,and well develop and utilize in the future.Under the different Salinity condition, after soaking seeds with VB1、VC、VB6、Vpp，we study on the their seed germination and the seeding growth(the root lengthand weights, The seedling heights and weights), it shows that: under differentsalinity, the seed germination and the seedling growth of any treatment are inhibited.With the increase of the concentration, the germination rate, Vi、Gi all had fallen; theroot length and weight, the seedling heights and weights steadily sank down. There are also have pronounced difference between all treatments with four differentvitamins.VB6、VC in all treatments are alleviative the salt damage, VB6 is easier tocause to put forth buds than VC, and it’s quantitative value is the highest in theultimate germination rate, in root and seedlings’ hight and weight. Though the VPP、VB1 are seems to inhibite its growth. Under the high concentration150mM Nacl, theultimate germination rate in all treatments are below the 40%, but VB6、VC’squantitative values in any experiments are higher than CK,while VPP lower thanCK.Then we study on the TTC reductive capacity of roots and the content of Polinein leaves, the result shows that between the different salinity, different vitamintreatments, different varieties of the wheat have discrepancy.along with theincreasing concentraion of the salinity（75mM，100mM，150mM），TTC reductivecapacity of roots decreases, the accumulation of the content of Poline in leaves havean upward trend. The increase of VB6’s treatment are obviously, VC comessecond,VPP is nearly come up with CK, changes a little. In TTC reductive capacity of roots’s reserch, VB6、VC are higher than CK at any time,VB1 is not palpable,VPP is lower than CK, makes negative affect on wheat. In addition, varieties of thewheats are remain different, no matter it shows promoting or inhibiting, all fourvitamins have moreobvious effects on CHY16 than CHY12, but the tendency of theeffection are the same. It is say that VB6、VC can help wheat to standwith the saultwell, and promot in growth，they are the better reagent to mix with the seed.