银杏种胚脱水敏感性及其生理研究

银杏（Ginkgo. Biloba.L），又俗称白果，是起源于中国的特有珍贵树种。本实验选用银杏种子和种胚为材料，研究其脱水过程中存活率、抗氧化酶活性、ABA等生理变化，并通过外源处理方式提高种胚脱水耐性，探讨银杏种胚脱水敏感性与抗氧化系统、ABA的关系，为银杏种质资源的长期保存提供一定依据。研究结果显示： 银杏种子和离体胚对脱水均较敏感，快速脱水后完整种子临界含水量40.3%，半致死含水量约为32%左右，离体种胚分别为28.2%和22%左右，初步认为银杏属顽拗型种子。经比较银杏整粒种子、离体胚快速脱水时含水量变化情况，发现离体种胚比整粒种子更耐脱水，完整种子脱水对内部种胚是一种慢速脱水。 种胚脱水过程中，含水量高于24.5% 时，丙二醛（MDA）含量基本不变，抗氧化酶活性增加，抗氧化酶防御机制起作用；当含水量低于24.5%时，MDA含量显著增加，抗氧化酶活性大幅度降低，防御机制无法消除过氧化产物的大量积累，造成细胞损伤，种胚存活率下降。因此银杏种胚脱水过程中，特别是脱水后期，抗氧化酶活性的迅速下降和脂质过氧化作用的加强与积累是造成存活率快速丧失、对脱水敏感的主要原因之一。 银杏属典型后熟种子，脱落后种胚需经历形态和生理发育过程，这一阶段脱水耐性在后熟7个月达到最大，ABA含量也不断积累，并在最耐脱水时期达到峰值，继续后熟脱水耐性减弱，ABA含量也迅速降低，可能与银杏种胚完成后熟转而进入萌发阶段有关。在种胚快速脱水过程中，ABA含量不断降低，与存活率显著正相关。银杏种胚在后熟过程中ABA的含量较低以及脱水过程中的不断降低，可能是造成种胚不耐脱水的另一部分原因。 通过外源ABA处理种胚后可明显提高其脱水耐性。ABA处理的种胚SOD活性升高，脱水后抗氧化酶活性（GR除外）被进一步激发，从而减少脂质过氧化伤害，降低细胞膜结构的破坏。这也更进一步证实了ABA和抗氧化系统在银杏种胚脱水过程中的重要作用。

大豆种子低温吸胀相关基因的分离与蛋白变化的研究

大豆 (Glycine max (L.) Meer.)是人们日常生活中不可缺少的食品，也是一种非常重要的油质、蛋白资源。目前根据大豆种子吸胀阶段对低温敏感性的不同，可将其划分成3种生态型：低温非敏感型、低温敏感型及中间型。对于低温非敏感型的种子来讲，4℃下吸胀24小时对其发芽率影响很小，而敏感型种子萌发率不超过5%。我国属于温带大陆性气候，大豆播种后由于温度波动而造成一部分种子不能萌发，最终导致减产甚至绝产的现象普遍存在。高产是育种工作的主要目标，提高逆境胁迫的适应能力是高产的前提和基础，所以从分子角度研究种子吸胀非常必要，一方面能够挖掘新的基因资源，另一方面为今后育种工作提供必要的理论依据。 本试验以此为立足点，低温吸胀非敏感型大豆品种 (Z22)为材料，利用cDNA-AFLP方法及蛋白质技术分离与低温吸胀相关的基因及蛋白，得到结果如下： 第一，本试验成功的分离出4个受低温诱导的基因，半定量RT-PCR方法进一步验证了这4个基因在种子吸胀24小时内受低温诱导。 第二，利用RACE方法成功的得到2个完整的全长基因，在NCBI数据库中查找后发现其中1个基因为新基因，命名为SCHI基因 (SCHI：Soybean chilling-induced gene)。SCHI全长为390bp，编码分子量大约为14.2KD的蛋白；另外一个基因是已知基因，其同源序列已经在其他的物种中得到分离。由于此基因与核糖体蛋白L34高度同源，所以把把这个基因命名为SOL34 (Soybean L34)。 第三，利用半定量RT-PCR方法对基因表达模型进行分析，结果表明：SCHI在种子低温吸胀18～24小时期间诱导表达量最高，而当种子低温吸胀24小时后转入常温下，其表达量在常温下18小时左右迅速下降；ABA (100μM)、PEG (30%，10000)及NaCl (250mM)能够诱导SCHI的表达，在诱导表达量上，ABA和PEG诱导效果最明显，而NaCl能够微弱的诱导此基因表达；对不同生态型的大豆品种而言，低温吸胀过程中，SCHI在非敏感型种子中的表达量高于敏感型种子，但非敏感型和中间型之间没有差别；另外，SCHI在大豆胚轴中是诱导型表达，在叶片和根尖中则是组成型表达。SOL34的表达在萌发前24小时内被低温诱导，但在不同生态型之间没有差别。SOL34在胚轴和根尖中受低温诱导，在叶片中是组成型表达。 第四，SCHI能够在原核生物中表达出相应蛋白，诱导表达蛋白的分子量在26-29KD，大约为理论值的2倍，说明在大肠杆菌中被表达的蛋白以2聚体形式存在。另外低温试验结果表明SCHI能够提高菌落忍耐短时间-20℃低温的能力。 第五，利用双元表达载体把SCHI转入拟南芥植株，经过低温、干旱和盐胁迫后，转基因植株的成活率均高于野生型植株。超表达SOL34的拟南芥植株降低了对低温的耐性；而抑制拟南芥中L34的表达反而提高了植株对低温的抗性。 第六，本试验利用蛋白质等有关试验检测了大豆种子低温吸胀时蛋白质发生的变化。从吸胀 (4℃和22℃下24h)后的大豆胚轴中成功鉴定出上调蛋白点25个，下调蛋白点15个。其中有参与能量代谢反应 (占10%，例如柠檬酸脱氢酶和苹果酸脱氢酶等)、细胞生长与分裂相关反应 (20%，例如LEA蛋白和种子成熟蛋白PM26)、胁迫反应 (10%，如乙醇脱氢酶)、种子宿命和贮藏蛋白 (20%，大豆球蛋白)等蛋白在此过程中发生了变化，暗示种子萌发前期低温吸胀过程中多种代谢发生变化。细胞生长变缓、能量代谢增强、胁迫代谢蛋白的高表达以及贮藏蛋白降解速度减慢等变化都有利于种子在吸胀过程中度过低温环境，为以后的生长作好准备。

干旱胁迫下草莓水分调控及光合特性研究

以盆栽草莓(Fragaria×ananassa)为材料研究了水分胁迫下克隆植物草莓母株和子株间的水分调控机制及其与碳同化、光系统II激发能分配的关系。实验材料分为匍匐茎连接和剪断两个大组，进行两步实验。第一步实验，对连接组和剪断组的所有母株控水，子株充分供水；4天后进入第二步实验，把连接组分为两小组，对其中一组充分供水子株开始控水，另一组保持不变。结果表明，土壤干旱引起母株叶片失水，并使其净光合速率和气孔导度显著降低。但是连接组中供水良好的子株能有效缓解缺水母株的水分胁迫。当供水良好的子株也开始受到干旱处理的时候，则会加剧与之相连母株的水分胁迫。受胁迫母株可以通过加强渗透调节能力和降低水势从相连子株获取水分。虽然土壤干旱会造成受胁迫母株叶片脱落酸（abscisic acid, ABA）含量的大幅度增加，但是与之相连子株的叶片ABA含量并没有增加；并且气孔导度与ABA变化趋势一致。因此，我们认为：(1)草莓母株和子株间的水分运输是由二者的水势差驱动的；（2）ABA不会通过匍匐茎在母株和子株间传递并影响相邻子株气孔导度；（3）在水分异质性较大情况下，生理整合可明显提高克隆系统的碳同化能力和光系统II激发能利用效率。 同时研究了水分胁迫对草莓叶片叶绿素荧光诱导动力学参数Fm的影响。结果表明，在水分胁迫初期, 活体草莓叶片失水萎缩、叶面积和叶片厚度减小，单位叶面积的叶绿素含量升高，此时叶绿素荧光动力学参数Fm上升；当水分胁迫进一步加剧，单位叶面积的叶绿素含量开始下降，但Fm没有随之下降。离体叶片测定则没有出现Fm上升这一过程，Fm随着单位叶面积叶绿素含量的下降而下降。叶片叠加实验证明，增加叶片厚度也可以使Fm上升。综上我们认为在干旱胁迫进程中，活体草莓叶片的荧光动力学参数Fm出现上升是由单位面积叶绿素含量和叶片结构的变化共同决定的。

以保卫细胞原生质体为实验系统的气孔生理研究进展

本文主要介绍了自GCPs分离技术建立以来,气孔生理研究中有关保卫细胞对K~+的吸收、释放机制;保卫细胞的能量供应、碳代谢及其调节;环境信号如光、CO_2、ABA等对GCPs的作用及其机制等方面取得的进展。

两亲性三嵌段共聚物在选择性稀溶液中的自组装及应用

本论文主要研究了ABA和ABC型两亲性三嵌段共聚物在选择性稀溶液中的自组装行为，得到了多种形态新颖、结构复杂多样的胶束，研究了这些复杂胶束的形成过程，探讨了影响胶束形态的各种因素并通过适当的方法对胶束形态进行调控。研究了聚乙烯基毗陡（P4VP）／聚苯乙烯（PS）三嵌段共聚物P4VP-b-PS-b-P4VP在二氧六环／水中的自组装行为，成功得到了嵌段共聚物环状胶束，并通过实验研究了环状胶束的形成过程。结果表明，胶束形态依赖于退火时间的长短。随着退火时间的延长，胶束结构从棒状过渡到环形结构。以实验结果为基础提出了环状胶束形成的新的机理，即囊泡塌陷形成环。同时，通过改变实验条件还得到了一些新型的环状复合结构，如环套环形、鸟状、哑铃形、戒指形、网络状等结构，并得到了与计算机模拟一致的结果。通过不同的方法对ABA型三嵌段共聚物（P4VP-b-PS-b-P4VP）在选择性溶液中的自组装胶束形态进行调控：首先，详细研究了P4VP-b-PS-b-P4VP在不同的共溶剂中的自组装行为。结果表明通过单纯改变共溶剂的性质可以很方便地调节胶束的形态，得到了球、棒、囊泡等结构。并用混合溶剂的方法得到了长度和直径可控的纳米线胶束。同时，通过加入第二种选择性溶剂（核层嵌段PS的选择性溶剂甲苯）的方法使形成胶束的核层嵌段在胶核中的伸展程度增加，从而使胶束形态发生转变。其次，研究了加入表面活性剂十五烷基苯酚（PDP）以构建分子间氢键来调节P4VP-b-PS-b-P4VP的胶束形态。结果表明，通过调节PDP的加入量可以使胶束形态发生从球到棒，到网络状、再到囊泡结构的转变。通过实验对比系统地研究了PDP的加入对胶束形态转变的影响，提出了相应的形态转变机理。再次，研究了不同分子量的嵌段共聚物之间共混及共聚物与均聚物共混对胶束形态的影响。结果表明加入亲油嵌段的均聚物对共聚物胶束形态影响非常明显，胶束形态与加入的均聚物的分子量及加入量直接相关。同时得到了一些新形态的胶束，如海绵状、笼子状等。共聚物共混的研究结果表明：通过两种不同分子量的共聚物共混可以得到这两种共聚物胶束的过渡态结构。用共聚物混合的方法还可以得到一些具有生物模拟性的胶束结构，如乌贼状、章鱼状等。这加深了人们对囊泡的形成机理及各种胶束形态之间形态转变的认识。P4VP-b-PS-b-P4VP通过在二氧六环／水中的自组装形成了囊泡，结果表明囊泡的尺寸依赖于初始状态下共聚物在共溶剂中的浓度及退火时间。除得到常规的球形囊泡外，还得到一些非球形囊泡，如长条形、三角形、项链形等囊泡结构。结合计算机模拟的方法研究了囊泡的形成机理，发现这些不同结构的囊泡的形成是由于初始状态下密度涨落所引起的。研究了实验中经常出现的各种胶束形态共存现象的原因，发现体系中亚稳态的存在是多形态共存的重要原因之一。通过聚苯乙烯一左聚乙烯基毗睫一左聚氧乙烯（PS一b一PZVP一b一PEO）在THF／水中的二次自组装首次得到了一种具有生物模拟性的巨大的节状蠕虫胶束（SWM）。研究发现，SWM是由重复单元盘状结构和丝状结构相连组成的。最令人吃惊的是这种SWM与自然界中的一些生命体如蛆叫、蛹、昆虫类的幼虫结构非常相似。通过对SWM形成过程中的中间态胶束结构的深入研究发现SWM是由球形胶束通过二次自组装形成的。SWM的形成过程可以分为三个阶段：ABC三嵌段共聚物先组装形成球形结构；这些球型结构粘连在一起形成梭形的中间结构；这些梭状中间结构中的球经过重组和重新调整各嵌段的排布最终形成SWM。用所得到的嵌段共聚物胶束为模板，采用无电沉积的方法成功制备了各种形态的金属一有机高分子纳米复合材料。用简单的方法还得到了导电金属金一银的双金属纳米结构材料。这些纳米结构材料在微电子器件等领域有潜在应用价值。以上研究结果丰富了人们对嵌段共聚物在选择性介质中自组装行为的理解，为人们提供了对生物材料自组装本质的理解的依据。这在两亲性分子在溶液中自组装的基础研究方面以及基于这些自组装形态而构建结构及功能更复杂的纳米结构材料等方面都有一定的意义。

电化学SPR研究及形貌可控的纳米结构合成

围绕论文题目“电化学SPR研究及形貌可控的纳米结构合成”，我们将SPR传感膜同时用作电化学研究的界面，开展了相关的EC-SPR研究工作。同时，在一些可控纳米结构的合成与表征方面进行了创新研究。 本论文研究工作的主要内容和创新点表现在以下几个方面：1．生物分子模板生长法为构建具有特定功能的新颖材料提供了新的途径。报道了一种基于固定的DNA为模板通过电化学途径合成DNA-聚苯胺复合物的方法。在这种条件下，目标生物分子能保持其天然结构和生物活性，能用于构建功能多样性的导电聚合物结构。2．首次用溶液中溶解氧现场原位还原产生的活性氧中间体作氧化剂，在蒸镀的金膜电极上阴极极化合成聚苯胺（PANI）。聚苯胺膜的厚度可很容易地在分子水平的尺度上控制，其表面形貌对金膜表面层原子的结晶取向非常敏感。在多晶金电极上可得到岛型的纳米结构，而在单晶Au（111）电极上则聚合得到超薄膜。3．在金电极表面电化学聚合形成的导电聚合物聚吡咯（PPy）膜被用作双层磷脂 膜（BLMs）的新的支撑体。PPy膜支撑的双层脂膜的形成依赖于所用脂分子的化学结构，在一定程度上PPy膜支撑的双层脂膜类似于传统的双层脂膜结构，在脂膜结构的内外两侧保持着水介质环境。PPy膜支撑的双层磷脂膜可很方便地用于仿生膜研究。4．采用光刻法构建SPR阵列传感器的金膜点阵列，拟将来用于SPR成像分析。把正型光刻胶旋涂于SPR金片表面，紫外光通过自制掩模曝光后，用碱液显 影。然后采用选择性化学刻蚀暴露出的金膜，最后用剥离液去掉未曝光的光 胶层，从而构建所需的金膜点阵列，点的大小和间距可方便地由掩模来控制。用壳聚糖为例进行了金膜点阵列的表面修饰与组装。点阵列间的玻璃表面能 抑制亲水性和疏水性分子的吸附，这在SPR成像分析及高通量筛选方面将非常有用。5．在生物学上，生物大分子或有机体通常能调节及控制生物／无机杂化材料和晶 体的形貌及组装，这个过程被称为生物矿化。我们报导了基于生物小分子，L一氨基酸的金纳米结构的生物合成。在没有表面活性剂及硬模板存在下，天冬氨酸能直接还原氯金酸生成大量的厚度小于30nm的金纳米盘，该纳米盘为单晶结构，主要晶面为｛111｝，特征形貌为平均边长为590nm的对称六角形以及平均边长为840nm的去顶角三角形纳米晶体。6．苯胺及其衍生物作为模型化合物被用于有意图地构建金属纳米材料。苯胺还原氯金酸生成金核直径38nm壳gnm的核／壳结构的球形纳米粒子，3-氨基苯甲酸（3-ABA）调制生成厚度为20nm边长为105nm的形状规则的金纳米片，4一氨基苯甲酸（4-ABA）指导生成直径为18nm长度为微米级的纳米线结构，2一氨基苯磺酸（2-ABS）能调节生成直径为13．7nln长度可达几十微米的“之”字形的金纳米线，而1-（4-氨基苯基）乙二胺-N，N，N'，N'-四乙酸（4-ABEDTA）能还原氯金酸并相应控制生成结构完好由平均直径为19nm的金球形纳米粒子连接的线结构并进一步组织成纳米分形网络结构，表明苯胺环上取代基的种类和位置对金属纳米结构形貌的调节有直接影响。

黄菠罗外植体复幼技术及其生理基础

本文以组织培养技术为手段，在成功实现黄菠罗微体繁殖与植株再生的基础上，通过对5个发育阶段初始材料的继代培养和继代过程中异戊烯基型细胞分裂素(iP型CKs)、玉米素型细胞分裂素(Z型CKs)、赤霉素(GAs)、生长素(IAA)、脱落酸(ABA)和多胺的连续检测分析，初步确定了黄菠罗幼-成龄阶段转化与五种内源激素、多胺含量之间的关系，探讨了黄菠罗成龄材料复幼复壮的可能性，确定了黄菠罗幼-成龄阶段转化的生物学指标，其研究结果可为黄菠罗无性系快繁和优良繁殖材料选择提供一定的理论依据，同时也为黄菠罗进一步开发利用打下基础。 主要结论： 1. 黄菠罗组织培养获得成功，优化培养基为 MS+2.5mg/l6-BA+0.25mg/LNAA，且约30年生 树木腋芽或一年生嫩枝可作为黄菠罗快速繁 育较适龄的外植体。 2. 成龄阶段黄菠罗可以获得部分复幼或复壮。 3. 在黄菠罗幼-成龄发育阶段转化的过程中， IAA、Z型CKs/总CKs、IAA/ABA、 Spd-S、Spm-S、DAP-SH、Spm-SH、Put- PH、Spd-PH、总Spd和总Spm逐渐 升高，而Z型CKs、iP型CKs、总CKs、iP型 CKs/总CKs、iP型CKs/GAs、iP型CKs/IAA、 GAs/IAA、iP型CKs/ABA、Z型CKs/IAA、Z型 CKs/GAs、Z型CKs/ABA、DAP-S、Put-S、总 PAs-S、Put-SH、Spd-SH、DAP-PH、Spm- PH、总Put、总PAs、DAP-S/ Spd-S、DAP- S/Spm-S、Put-S/Spd-S、Put-S/Spm-S、 DAP-SH/ Spm-SH、DAP-S/(Spd-S+Spm-S)、 Put-S/(Spd-S+Spm-S)、Put-SH/Spm-SH、 (DAP-S+Put-S)/(Spd-S+Spm-S)和(DAP- SH+Put-SH)/(Spd-SH+Spm-SH)逐渐降低；在 整个继代过程中，Z型CKs、iP型CKs、总 CKs、Gas、IAA、IAA/ABA、iP型CKs/总 CKs、iP型CKs/GAs、Put-S、 Spd-S、总PAs-S、Put-SH、Spm-SH、DAP- PH、Spd-PH、总Put、Put-S/Spd-S、 Put-S/Spm-S、Put-SH/Spd-SH、Put- SH/Spm-SH、Put-S/(Spd-S+Spm-S)、Put- SH/(Spd-SH+Spm-SH)、(DAP-S+Put-S)/ (Spd-S+Spm-S)、(DAP-SH+Put-SH) /(Spd-SH+Spm-SH)和总PAs-S/总PAs-SH逐 渐升高，而ABA、Z型CKs/总CKs、iP型 CKs/IAA、GAs/IAA、Z型CKs/IAA、Z型 CKs/GAs、Z型CKs/iP型CKs、GAs/总CKs、 Spm-S、DAP-SH、Spd-SH、总PAs-SH、Put- PH、Spm-PH、总PAs-PH、总Spd、总Spm和总 PAs逐渐降低。 4. 培养物中5种内源激素代谢表明，Z型CKs/iP 型CKs、iP型CKs/总CKs的比值和GAs含量可 以作为黄菠罗幼-成龄阶段转化的指标，而 iP型CKs/GAs、iP型CKs、总CKs、GAs、iP型 CKs/IAA和IAA/总CKs则是成龄阶段黄菠罗复 幼的主要原因，且对复幼的影响依次降低。 5. 培养物内源多胺代谢表明，游离态和酸可溶 性Put/Spd和Put/( Spd+Spm)、Put-S/Spm- S、和总PAs-S/总PAs-SH可以作为幼-成龄阶 段转化的指标,而总PAs-S则是对成龄黄菠罗 复幼或幼龄黄菠罗保持较高分裂分化能力的 主导因素，依次是总PAs-S、Spd–S、总PAs- S/总PAs-SH、总PAs-SH、Put-S、Put-SH、 (DAP-S+Put-S)/(Spd-S+Spm-S)、Put-S/( Spd-S+Spm-S)、Put-SH/Spm-SH、Put- S/Spd-S。

植物激素对土壤污染早期诊断研究

通过菲污染诱导动态响应预实验确定了小麦为供试植物，赤霉素(GA3)、细胞分裂素的玉米素和玉米素核苷总含量(Z&ZR)以及脱落酸(ABA)为植物激素供试指标，并且确定试验终点为小麦生长14d。在预实验的基础上，以草甸棕壤为供试土壤，采用室内培养箱盆栽方法，进行多环芳烃菲、芘、荧蒽和苯并(a)芘单一，两-两复合以及三-三复合与3种植物激素的剂量-效应关系研究；3种植物激素对土壤重金属镉、铅、铜和锌单一污染诱导的响应关系研究。并以多环芳烃类的菲和重金属类的镉为代表污染物，进行小麦芽期生长指标及植物激素测定，研究植物激素与生长指标对污染物的响应及敏感性比较。 多环芳烃单一及复合污染诱导实验结果表明：ABA对菲、芘、荧蒽、苯并(a)芘单一及复合污染诱导的响应趋势一致，即随着污染诱导浓度增加，ABA含量显著增加；GA3与四种多环芳烃单一及复合污染诱导存在一定的响应关系，但响应趋势呈多样性变化；对于多环芳烃单一及复合污染诱导，随着污染物浓度增加，Z&ZR含量出现了先增加后减少的趋势，表现为显著的污染诱导抑制效应。ABA和Z&ZR对多环芳烃污染诱导响应敏感，这为其成为土壤污染诊断指标提供了可能性。 在不同的镉、铅、铜、锌污染处理中，小麦体内ABA含量均出现明显增加趋势，与多环芳烃污染胁迫响应一致；GA3含量对镉、铅、铜、锌四种重金属污染胁迫响应趋势相同，均为先增加后减少。对于土壤镉和铅污染胁迫Z&ZR表现为先增加后减少的响应趋势，而对于铜和锌污染胁迫，Z&ZR含量变化与镉、铅胁迫时不同。 植物激素与生长指标对污染胁迫的响应及敏感性比较研究表明，对多环芳烃菲污染诱导，植物激素含量同生长指标相比具有显著的敏感性。对重金属镉污染诱导，内源激素含量和生长指标敏感性都显著，小麦根和芽中ABA含量显著增加，进一步证明了其在植物污染抗逆中的作用机理。

土壤镉污染的遗传及生态毒性研究

针对蚕豆根尖微核试验方法现存的问题并结合土壤自身的特性，进行了蚕豆微核试验方法改进的试验研究。在此基础上，本文选择蚕豆幼苗次生根根尖为供试材料，以细胞有丝分裂指数（MI）、染色体畸变率（CAF）和微核率（MNF）为试验指标，开展了我国典型土壤（红壤、棕壤和黑土）中低剂量（0~10 mg•kg-1）镉污染遗传毒性剂量-效应关系研究。研究针对蚕豆根尖微核遗传毒性评价指标“污染指数”的缺陷，首次提出“毒性指数”。“毒性指数”将有丝分裂指数、染色体畸变率和微核率指标所得数据结合，评价土壤镉遗传毒性水平。与此同时，研究选择蚕豆幼苗叶片为供试材料，进行了以抗氧化酶活性（SOD、POD和CAT）和植物激素含量（ABA、GA3和Z&ZR）为指标的镉污染胁迫生态毒理试验研究，并比较了上述三类指标对土壤镉污染的毒性响应及敏感性。 蚕豆根尖微核方法改进试验结果表明，在三种备选试验材料中，蚕豆初生根、次生根根尖细胞和芽尖细胞均对镉胁迫响应显著。与芽尖细胞相比，根尖细胞MI、CAF和MNF之间具有较好的相关性；而次生根尖细胞比与初生根尖细胞对镉响应更敏感，遗传毒性效应显著相关。在上述方法学研究基础上，将蚕豆幼苗次生根根尖微核试验应用到典型土壤镉污染的研究中，结果表明，同一剂量下，红壤中镉的遗传毒性效应最强，棕壤次之，黑土最不显著。土壤pH值与有机质含量对镉的遗传毒性影响较大。 抗氧化防御酶活性试验结果表明，POD和SOD在0~10 mg•kg-1镉浓度范围内先升高后降低，对镉毒性的指示效果最明显；CAT反应与SOD、POD相反，其作用机理有待进一步探讨。植物激素试验结果表明，ABA和Z&ZR对镉胁迫响应敏感，其含量与镉浓度存在良好的剂量-效应关系；GA3随着镉浓度的增加表现出无规律性，表明它对污染物的响应具有选择性。 试验数据显示，不同指标对镉胁迫均具有响应，但响应的域值及敏感度不同。比较而言，蚕豆根尖微核遗传毒性指标指示效应更明显。将各指标联合使用可使土壤镉污染的遗传和生态毒性诊断更为全面、有效。本研究为土壤镉污染早期诊断敏感指标筛选及指标组合提供了实验依据，为土壤重金属污染早期诊断奠定试验和理论基础。

干旱胁迫下沙地樟子松根部信号及其生理响应

土壤干旱是沙地樟子松生长发育的主要限制因子。论文采用PEG处理和盆栽干旱处理模拟沙地樟子松幼苗对干旱胁迫的生理生态响应。通过对水势、根部及叶片ABA含量、气孔导度、蒸腾速率、净光合速率同步测定，并分析水势、ABA含量变化与生理响应关系，同时对脯氨酸、叶绿素含量、束缚水、自由水进行测定。得出以下几点结论：（1）无论是PEG处理还是盆栽干旱处理，水势在处理后期与所孔导度、蒸腾速率、净光合速率都具有一定相关性，因此，植株水分状况仍是植株调控自身生理变化的一个途径，樟子松可利用高的渗透势适应干旱胁迫。但水势变化落后于ABA变化说明除水势外，还存在其它信号参与生理调控。（2）干旱胁迫下，根部和叶片ABA均可迅速积累，但叶片ABA较根部滞后。根部ABA可通过腾流运输到叶片从而降低叶片所孔导度、蒸腾速率以达到减少水分损失的目的，干旱处理条件下生理变化是由根部ABA含量变化向气孔传达了根部缺水的信号，并由ABA诱导气孔导度减小。（3）1%土壤含水量可能是沙地樟子松幼苗干旱胁迫的临界值。低于此临界值，幼苗生长受到限制。（4）电导率在干旱胁迫初期略下降但不明显，然后上升。（5）在干旱胁迫条件下脯氨酸增加，叶绿素减少，自由水减少，束缚水增加。（6）干旱协迫信息感知与传递是多种胁迫信号共同作用的结果。

玫瑰花芽分化期矿质元素和内源激素的变化规律

本文以沈阳市于洪区玫瑰种植示范区内多季花冷香玫瑰和单季花平阴玫瑰为研究对象，通过野外采样并测定了两者在花芽分化期矿质元素（P、K、Ca、Mg、Fe、Zn、B、Mn、Cu）和内源激素（IAA、GA、ZR、ABA）的含量，并对其含量及比值变化进行分析，初步确定了玫瑰花芽分化与元素、激素之间的相互关系。研究结果将为玫瑰花期的化学调控、提高产花量和正确制定栽培技术措施提供理论依据，也为玫瑰进一步开发利用打下基础。主要结论：1．在冷香玫瑰和平阴玫瑰的整个花芽分化期，P、K、Cu、Zn大体上均呈下降趋势，表明它们可能参与玫瑰成花；ca、Mg在冷香玫瑰和平阴玫瑰的花蕾形成期含量都下降，可能意味着ca、Mg参与两者的花蕾形成；Fe与玫瑰花芽分化关系不大；高含量的B和Mn有利于两者花蕾形成。2．在冷香玫瑰和平阴玫瑰花蕾形成期，IAA和GA出现低水平，ZR和ABA出现高水平，激素比值（ABA/GA，ABA／IAA，ZR/GA，ZR/IAA）出现高水平，表明ABA和ZR促进两者开花；而IAA和GA抑制两者开花；高比例的（ABA/GA，ABA/IAA，ZR/GA，ZR/IAA）有利于两者开花。

青杨组(Populus section Tacamahaca Spach)不同种对增强UV-B的反应差异

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。本项目以青杨组杨树为模式植物，从形态和生理方面研究了来自不同UV-B背景下的康定杨与青杨在增强UV-B下的反应及其反应差异,并探讨了干旱、施肥对它们抗UV-B能力的影响。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，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。主要研究结果有以下： 1. 在温室中经过增强UV-B处理，杨树的外部形态及生理活动受到了一定程度的影响。增强UV-B导致康定杨、青杨的生物量、叶面积及节间长度降低，叶片增厚，SOD活性升高，膜伤害增加，而对叶片数目、R/S、叶绿素A、叶绿素B及整个叶绿素含量没有影响。两种杨树对UV-B胁迫的响应存在差异：在增强UV-B条件下，青杨的植株高度、生物量、叶面积、脯氨酸含量、长期用水效率受到的影响大于康定杨，相比而言，康定杨在比叶面积、叶片厚度、可溶性糖含量、UV-B吸收物质的含量及SOD和GPX活性方面增加的程度大于青杨。这些区别说明，来自于高海拔的康定杨比来自于低海拔的青杨对增强UV-B 具有更强的耐性。我们认为二者在叶片厚度、比叶面积、UV-B吸收物质含量及SOD、GPX活性差异是导致对增强UV-B耐性不同的原因。 2. 干旱与增强UV-B对杨树的生长和生理特性均产生了影响，而且两种胁迫共同作用时干旱表现减弱或加剧了UV-B对杨树某些形态和生理特性的影响。 据试验结果，干旱显著地降低了杨树的株高、叶片数目、叶面积，增加了叶片厚度，促进ABA的积累，提高了CAT活性。对于干旱，两种杨树之间也表现出了一定的差异性。可溶性蛋白质和脯氨酸在青杨叶片中得到显著积累，而在康定杨中没有变化。此外，CAT、长期用水效率在康定杨中受到的影响更加明显。长期用水效率的不同变化趋势说明两种杨树对水分胁迫采用了不同的用水策略，康定杨采用的是节水用水策略，提高用水效率，而青杨采用的是耗水的用水策略。根据干旱对叶面积、脯氨酸、ABA含量、CAT活性及长期用水效率等方面的影响，我们认为来自高海拔地区的康定杨比来自低海拔的青杨有更大的耐旱性，这是对生长环境长期适应的结果。在高海拔地区，因霜冻常带来土壤水分不可利用，降低了根系对水分的吸收，树木容易受到的生理性干旱。另外，高海拔的地区低的气温使植株对严寒有较强的耐性，减少了水分的需要。 生长于增强UV-B下的康定杨和青杨植株表现为高度降低，叶面积缩小，比叶面积增加；叶片栅栏组织、海绵组织均受到增强UV-B的影响，其厚度的增加导致整个叶片变厚。增强UV-B还显著提高了杨树的APX活性、UV-B吸收物质含量，而对叶片数目、ABA、可溶性蛋白质含量及CAT活性没有产生影响。试验中也观察到了两种杨树对增强UV-B响应的差异：与康定杨相比，在增强UV-B下青杨株高、叶面积降低的程度更大一些，SOD活性显著提高。另外UV-B吸收物质受到的影响不同。根据这些差别，高海拔的康定杨(3500 m)比来自低海拔的青杨(1500 m)增强UV-B有较强的耐性。 与水分充足情况下UV-B对植株的影响相比，干旱对杨树抗增强UV-B产生了一定的影响，表现为加剧或减弱UV-B对植物的影响，但这种影响与形态、生理指标有关。当干旱与增强UV-B共同作用时，杨树植株的株高、叶面积进一步降低、叶片进一步增厚。就脯氨酸的积累的而言，在没有水分胁迫时，增强UV-B促使它显著增加，而在干旱处理下这种效果变得不明显。干旱对增强UV-B的影响还与杨树的种类有一定的关系。在康定杨中，干旱减弱了增强UV-B对栅栏组织与海绵组织的影响，且在植株高度、叶面积上表现出累加效应，而在CAT上交互作用显著；但在青杨中干旱则加剧增强UV-B对栅栏组织与海绵组织的影响，在植株高度、叶面积及比叶面积上表现出显著的交互作用。据碳同素分析，在水分充足的条件下，无论是康定杨，还是青杨，增强UV-B均导致其长期用水效率的提高，然而当两种胁迫共同作用时，长期用水效率则表现出差异，在青杨中，长期用水效率得到进一步增高，而康定杨中干旱的效应被增强UV-B所减轻。 3. 田间试验表明，杨树的生长、生理特征都受到养分和增强UV-B的影响。施肥对杨树的影响表现为：提高了叶面积、生物量及SOD的活性，降低了抗坏血酸含量。对于施肥作用，两种杨树的反应也有区别：在康定杨中施肥显著增加了的叶片长度、宽度及光合色素的含量，降低了净光合速率、气孔导度及胞间CO2浓度；在青杨中，则SOD、GPX、APX活性表现增加。从试验看出，施肥对来自于高海拔地区的康定杨(3500 m)的影响较大，对来自低海拔的青杨(1500 m)影响较小，这与它们对原产地的生境适应有一定关系。在康定杨生长的高海拔地区，低温度和湿度不能为地上凋落物或土壤中的根分解提供理想的条件，造成当地土壤的低养分状况，所以当肥料施用以后，效果显著。 经过增强UV-B处理，杨树叶片中UV-B吸收物质含量、GPX的活性得到提高，而脯氨酸、丙二醛、可溶性蛋白质、叶绿素及类胡萝卜素含量没有受到影响。对于增强UV-B两种杨树受到的影响也有所不同：在青杨中增强UV-B导致叶面积缩小，生物量、净光合速率降低，APX的活性及长期用水效率的提高，而对康定杨的这些指标没有产生显著影响，相反抗氧化酶的活性明显高于青杨。这些差异性是由于两种杨树对原产地不同UV-B背景的长期适应结果。康定杨长期生长在较高UV-B环境中，对UV-B有较强的耐性。而青杨适应于较低的UV-B环境，对增强UV-B较为敏感。 试验中施肥也影响了植株对增强UV-B的反应，不过这种影响与杨树的种类及测定指标有一定的相关性。例如，在缺肥的情况下，青杨的长期用水效率和康定杨的叶绿素含量都受到增强UV-B的显著影响，而施肥以后这种影响变得不显著。在缺肥的条件下，GPX、APX在青杨中的活性、GPX在康定杨中的活性对增加UV-B反应不敏感；而施肥以后则变化显著，同样胞间CO2浓度在康定杨也有类似的变化。 For past decades, 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. In this experiment, different species of Populus section Tacamahaca Spach from different UV-B background were selected as a model plant to assess the effects of enhanced UV-B radiation. Morphological and physiological traits induced by enhanced UV-B were observed and the different responses between P. kangdingensis and P. cathayana were discussed, furthermore the influences of drought and fertilizer on responses induced by enhanced UV-B were studied. Since poplars play an important role in lumber supply, and are important component of ecosystems due to their fast growth and wide adaptation, the study could provide a strong theoretical evidence and scientific direction for the afforestation, and rehabilitation of ecosystem. The results are as follows: 1. The experiment conducted in a greenhouse indicated that morphological and physiological traits of two poplars were affected by enhanced UV-B radiation. Enhanced UV-B radiation not only reduced biomass, leave area and internode length, but also increased leaf thickness and SOD activity as well as MDA concentration and electrolyte rate. However, no significant changes in leaf numbers, root shoot ratio, and total chlorophyll and chlorophyll component were observed. There were different responses to enhanced UV-B radiation between two species. Compared with P. kangdingensis, cuttings of P. cathayana, exhibited lower height increment and smaller leaf area. In addition, there were significant differences in free proline, soluble protein, and UV-B absorbing compounds, and the activity of SOD and GPX, long-term WUE between them. Differences in plant height, biomass, leaf area, free proline concentration, and long-termed WUE showed that P. cathayana were more affected by enhanced UV-B radiation than P. kangdingensis. In contrast, more increase of specific leaf mass, leaf thickness, and soluble sugar, and UV-B absorbing compounds, and activity of SOD and GPX were observed in P. kangdingensis. According to these results, we suggested that P. kangdingensis from high elevation, which adapted to higher UV-B environments, had more tolerance to enhanced UV-B than P. cathayana from low elevation, which adapted to lower UV-B environment. We believe it was the difference of leaf thickness, specific leaf mass, and UV-B absorbing compounds as well as the activity of SOD and GPX resulted in lower adaptation of P. cathayana to enhanced UV-B radiation. 2. Growth and physiological traits of two poplars were affected by both drought and enhanced UV-B radiation. Moreover, it was observed that when two stresses applied together drought could exacerbate UV-B effects or decrease sensitivity to UV-B. In the experiment, drought significantly decreased plant height, leaf numbers, leaf area, and increased leaf thickness, and ABA, and CAT activity of two poplars. There were significant interspecific differences to drought stress. Exposed to drought, soluble protein and proline concentration were increased in P. cathayana but not in P. kangdingensis. However, more changes in CAT and long-term WUE were observed in kangdingensis. Different change in long-term WUE suggests that two poplars adapted different water-use strategies. P. kangdingensis employ a conservative water-use strategy, whereas P. cathayana employ a prodigal water-use strategy. Based on the differences in leaf area, accumulation of free proline and ABA, CAT activity as well as long-term WUE, we believed that P. kangdingensis from high elevation had a greater tolerance to drought than P. cathayana from low elevation，which is the result of adaptation to local environment. In high elevation area, trees are prone to suffer from physiological drought because of un-movable water caused by frost. Besides lower temperature enable the plants had greater adaptability to frost as a results the requirement of water is reduced Enhanced UV-B radiation decreased shoots height, leaf area, and increased specific leaf mass and thickness of palisade and sponge layer as well as APX activity and UV-B absorbing compounds in both species. Whereas, leaf numbers, ABA content, soluble protein and CAT activity showed no differences to enhanced UV-B radiation. Interspecific differences were also observed. Compared with P. kangdingensis, P. cathayana showed lower shoot height and smaller leaf area, higher SOD activity. Besides, variation in UV-B absorbing compounds was found. These differences suggested that P. kangdingensis from high elevation (3500 m) was more tolerant to enhanced UV-B radiation than P. cathayana from low elevation (1500 m). Compared with morphological and physiological changes induced by enhanced UV-B radiation under well-watered conditions, drought exacerbated or decreased these changes. However, these effects vary with parameters measured. When two stresses applied together, shoot height and leaf area further decreased while leaf thickness further increased. Under well-watered conditions, enhanced UV-B radiation significantly increased proline content, but such effect was not observed under drought conditions. The effect of drought on enhanced UV-B radiation was related to species. For example, drought reduced the effects of enhanced UV-B radiation on palisade parenchyma and sponge mesophyll in P. kangdingensis, and additive effects in shoot height and leaf area and interactive effect CAT activity were observed. In contrast, for P. cathayana drought significantly exacerbated the effects of enhanced UV-B radiation on palisade parenchyma and sponge mesophyll; there were noticeable interaction in shoot height, leaf area and specific leaf mass. As far as long-term WUE is concerned, it was increased by enhanced UV-B radiation under well-watered conditions in both species. While different effect was observed between two species in combination of two stresses. Long-term water use efficiency was further increased in P. cathayana whereas the effect was less significant in P. kangdingensis. 3. The field experiment showed that growth and physiological traits of poplars were affected by nutrition and enhanced UV-B radiation. Fertilization significantly increased leaf area, biomass and SOD activity, reduced Ascorbic acid concentration. There was interspecific difference in response to fertilization. For P. kangdingensis, fertilization significantly increased leaf width, leaf length and photosynthetic pigments content while net photosynthetic rate and stomatal conductance, intercellular CO2 concentration were significantly decreased. However, for P. cathayana, these parameters were unaffected except the increase of SOD, GPX and APX activity. From above, it could concluded that P. kangdingensis from high elevation was more affected by fertilization than P. cathayana, This difference was due to adaptation to local environment., The low temperature and moisture where P. kangdingensis was collected can not provided optimum to decompose roots and litter fall as a result the nutrition in soil was poor. Exposed to enhanced UV-B radiation, for both species UV-B absorbing compounds and GPX activity were significantly increased while proline, MDA, soluble protein, chlorophyll, carotenoids were not affected. Different responses were also observed between the two species. Enhanced UV-B radiation caused significant decreases in leaf area, biomass, net photosynthetic rate and increase in APX activity and long-term WUE in P. cathayana but not in P. kangdingensis. In addition, activity in antioxidant enzymes was much higher in P. kangdingensis than in P. cathayana. In the experiment fertilization affected responses of cuttings to enhanced UV-B radiation, but it concern species and parameters measured. Long-term WUE in P. cathayana and chlorophyll in P. kangdingensis were significantly increased by enhanced UV-B radiation under non-fertilization treatments while the increase was not found under fertilization treatment. In contrast, under no fertilization treatment enhanced UV-B radiation did not affected GPX and APX activity in P. cathayana and GPX in P. kangdingensis while significant increase appeared after application of fertilization. Similar effect of enhanced UV-B radiation on intercellular CO2 concentration in P. kangdingensis was observed.

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.

沙棘(Hippophae rhamnoides L.)在低温和增强UV-B胁迫下的生态生理反应

植物生长和生产力受到自然界各种形式的生物和非生物胁迫因子的影响。这些胁迫包括低温、高温、盐碱、干旱、洪水、重金属、虫害、病害和紫外线辐射等等。而人类活动大大加剧了这些胁迫所带来的影响。由于人类污染而导致臭氧层衰减以及由此产生的地球表面紫外辐射增强已经成为全球气候变化的一个主要方面。UV-B胁迫，甚至当前的辐射水平，所带来的影响已经引起科学工作者的广泛关注。 为了生存和繁殖，植物不得不面临环境中各种潜在胁迫所带来的负面影响。然而，植物生活型的不可移动性决定了其逃避胁迫的局限性。因此，绝大多数植物都是通过对胁迫作出反应，通过修复或者更新组织来降低伤害。而植物应对环境变化的能力则是由其生长模式的种属特异性和本身的遗传组成所决定。在自然界，植物常常同时面临多种胁迫，这些胁迫所引发的植物反应可能具有叠加、协同或者拮抗作用。沙棘是一种具刺、具有固氮功能的多年生雌雄异株灌木，广泛分布于亚欧大陆的温带地区和亚洲亚热带的高海拔地区。在中国，沙棘常常被用作植被恢复中的先锋树种而大量栽培。本文采用沙棘作为模式植物，试图探索木本植物对低温，UV-B辐射增强以及其与干旱的复合胁迫的响应以及沙棘对这些胁迫响应是否具有种群差异性。 对来自南北两个种群的沙棘进行短日照和低温处理，检测了其在抗寒锻炼和抗寒性发育过程中存在的性别差异。结果表明，短日照和低温都分别能够诱导抗寒锻炼的发生，而两者同时存在对所有实验植株抗寒性的大小具有叠加效应。然而，短日照和低温所诱导的抗寒性在两个种群中都具有性别差异性，雄性植株比雌雄植株对短日照和低温更为敏感。同时，南北种群间也存在差异性，北方种群的植物比南方种群的植物对短日照和低温敏感，从而在短日照下抗寒锻炼的发生时间更早，低温诱导的抗寒性更大。短日照和低温诱导植物增加抗寒性的同时伴随着脱落酸的变化。脱落酸的变化因处理，种群和性别的不同而不同。这些生理反应表明不同的沙棘种群，不同的植株性别对同一环境胁迫可能存在不同的生存策略。 比较了来自高低两个海拔的沙棘种群对于干旱和UV-B辐射增强以及两者复合胁迫条件下的生理生态反应。干旱使两个种群中植株总的生物量，总叶面积，比叶面积，叶片含碳量，含磷量，木质素含量和碳氮比显著降低，使根冠比，粗根细根比和叶片脱落酸含量显著增加。干旱而非UV-B使得δ13C 值显著增加。但是，比较而言，来自高海拔的种群对干旱反应更为强烈，而来自低海拔的种群对UV-B更敏感。在UV-B辐射增强的处理下，干旱所诱导的脱落酸的积累被显著抑制。而且我们检测到在一些指标上存在显著的干旱×UV-B交互作用，如两个种群中在总生物量上，低海拔种群中在总叶面积，粗根细根比上，高海拔种群中在比叶面积，δ13C值，木质素含量上都存在明显的交互作用。这些结果表明这两个种群对胁迫具有不同的适应性反应，来自高海拔的种群比来自低海拔的种群更能够抵御干旱和UV-B胁迫。 室外实验表明，UV-B 去除/增补对沙棘高低两个海拔种群的影响都不大。对生物量的积累，植株高度以及一些常见的胁迫反应生理指标比如丙二醛、ABA 和游离脯氨酸都没有显著影响。UV-B 的效应比UV-A 大，植物反应在无UV 和仅有UV-A 的处理间没有什么区别。然而，UV-B 去除的两个处理和UV-B 存在的两个处理间存在显著区别。UV-B 使得两个种群都显著降低了比叶面积(SLA)，但却使长期用水效率增加。但UV-B对光合色素和光合系统II 的影响不大。总体看来，来自低海拔的种群对UV-B 更为敏感。 Plant is adversely affected by various abiotic and biotic stress factors. These stressors includelow temperature, heat, salt, drought, flooding, heavy metal toxicity, wounding by herbivores,infecting by pathogenic microorganisms, ultraviolet (UV) radiation and so on. Variousanthropogenic activities have accentuated the existing stress factors. One of the mostimportant aspects of global change is that of stratospheric ozone depletion caused by seriousanthropogenic pollution and the resulting increase in UV radiation reaching the surface of theEarth. Scientists have become concerned about the effects that considerable UV-B stress, evenat current levels. In order to survive and reproduce, plants have to be able to cope with lots of potentiallyharmful stress factors that are almost constantly present in their environment. Most plants’responses under stress are to neutralize the stress, repairing the damage or regrowing newtissue rather than to avoid it due to their sessile life style. The plant defense capacity dependson plant-specific modular growth patterns and genetic make-up that allows for flexibleresponses to changing environments. Plants usually encounter several stresses simultaneouslyunder field conditions, and the stresses may cause a variety of plant responses, which can beadditive, synergistic or antagonistic. Sea buckthorn (Hippophae rhamnoides L.), a thorny nitrogen fixing deciduously perennialshrub, which is widely distributed throughout the temperate zones of Asia and Europe and thesubtropical zones of Asia at high altitudes. It has been widely used in forest restoration as thepioneer species in China. In this paper, we used sea buckthorn as a model, tried to get some understand of how plants fight low temperature, enhanced UV-B radiation level and thatcombination of drought. And also, want to know whether does there exist some populationspecific responses to such stressors. Sexual differences in cold acclimation and freezing tolerance development of two contrastingsea buckthorn (Hippophae rhamnoides L.) ecotypes from northern and southern regions inChina were recorded after exposure to short day photoperiod (SD) and low temperature (LT).The results demonstrated that cold acclimation could be triggered by exposing the plants toSD or LT alone, and that a combination of both treatments had an additive effect on freezingtolerance in all plants tested. However, development of freezing tolerance was dependent onthe sex of plants under SD and LT, the males were clearly more responsive to SD and LT thanthe females in both ecotypes studied. On the other hand, development of freezing tolerancewas also ecotype-dependent, the northern ecotype was more responsive to SD and LT than thesouthern ecotype, resulting in earlier cold acclimation under SD and higher freezing toleranceunder LT. Moreover, development of freezing tolerance induced by SD and LT wasaccompanied by changes in ABA levels. These alterations in ABA levels were different indifferent treatments, ecotypes and sexes. Therefore, the differences in SD and LT-inducedphysiological responses showed that the different ecotypes and the different sexes mightemploy different survival strategies under environmental stress. Two contrasting populations from the low and high altitudinal regions were employed toinvestigate the effects of drought, UV-B and their combination on sea buckthorn. Droughtsignificantly decreased total biomass, total leaf area, specific leaf area，leaf carbon (C),phophous (P), lignin content and the ratio of C: N in both populations, and increasedroot/shoot ratio, fine root/coarse root ratio and abscisic acid content (ABA), in bothpopulations. Drought but not UV-B resulted in significantly greater carbon isotopecomposition (δ13C) values in both populations. However, the high altitudinal population wasmore responsive to drought than the low altitudinal population. The drought-inducedenhancement of ABA in the high altitudinal population was significantly suppressed in thecombination of drought and elevated UV-B. Moreover, significant drought × UV-B interactionwas detected on total biomass in both populations, total leaf area and fine root/coarse root inthe low altitudinal population, specific leaf area, δ13C value and leaf lignin content in the high altitudinal population. These results demonstrated that there were different adaptive responsesbetween two contrasting populations, the high altitudinal population exhibited highertolerance to drought and UV-B than the low altitudinal population. A field experiment was conducted to investigate effects of UV-B exclusion/supplementationon two altitudinal populations of sea buckthorn. UV-B exclusion or supplementation had littleeffects on both populations investigated. For instance, the total biomass, plant height andsome physiological index such as Malondialdehyde (MDA), ABA and free proline were notchanged significantly. The UV-B effects are more significant than that of UV-A, nodifferences were found between treatments of excluded UV and excluded UV-B. However,compared with treatments of UV-B exclusion (including absent of UV-B and all UV band),the present of UV-B (including near ambient environment and enhanced UV-B) significantdecreased specific leaf area, and increased long time water use efficiency as evaluated by δ13Cvalue. UV-B had little effects on photosynthetic pigments and Photosystem II (PSII). The lowaltitude population is more sensitive to UV-B than that of the high altitude population.

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.