大气CO2浓度升高对红桦碳水化合物含量和分配的影响

碳水化合物按其存在的形式可分为结构性碳水化合物和非结构性碳水化合物两种。前者主要用于植物体的形态建成；后者是参与植物生命代谢的重要物质。迄今为止，有关CO2浓度升高对植物叶片中的碳水化合物含量的研究较多，而对其它器官中碳水化合物含量以及碳水化合物在植物体内的分配响应研究较少。碳水化合物含量在植物各器官中的变化能够反映光合同化产物在叶和茎、枝和根中的转运情况；碳水化合物的分配与植物的生长模式相关，它的变化会对植物的生长情况产生影响。因此，为全面认识植物生理生化与生长过程对大气CO2浓度升高响应情况，需要对CO2浓度升高条件下植物体内碳水化合物的含量及分配变化进行深入的研究与探讨。本文应用自控、独立、封闭的生长室系统，研究了红桦幼苗根、茎、叶和枝的碳水化合物含量以及分配格局对大气CO2浓度升高(环境CO2浓度+350 µmol·mol-1) 的响应。研究结果表明：1) CO2浓度升高使红桦幼苗叶片中的非结构性碳水化合物含量显著增加。这可能会对光合作用造成反馈抑制，降低光合速率。2) CO2浓度升高使红桦幼苗根、茎和枝中的还原糖、蔗糖、总可溶性糖、淀粉和总的非结构性碳水化合物(TNC) 含量显著增加。说明CO2浓度升高促进了碳水化合物由叶片向枝、茎和根中的运输转移，支持了Finn和Brun的假设。3) 在总的非结构性碳水化合物(TNC) 中，淀粉所占比例最大。同样地，CO2浓度升高使TNC含量增加的部分中，淀粉所占的比例也最大。在叶片、枝、茎和根中淀粉含量增加部分占TNC含量增加部分的91.45%、88.23%、83.23%和82.01%。4) CO2浓度升高使红桦幼苗根、茎、叶和枝内的纤维素含量有增加的趋势，但未达到显著水平。需要进一步研究长期CO2浓度升高下，纤维素含量的响应程度。5) CO2浓度升高使碳水化合物在红桦幼苗体内的分配发生了改变。红桦幼苗体内碳水化合物分配变化的一致趋势是由地上部分向地下部分分配转移。其中，测定的所有碳水化合物均向根中分配增多。同时，CO2浓度升高使红桦幼苗的根冠比显著增大；根系干重显著增加。这些结果支持了Gorissen 和Cotrufo的假设，即碳水化合物向根中分配增多是根冠比增大的主要原因。6) CO2浓度升高使红桦幼苗体内的氮含量显著下降。氮含量的下降可能主要是由生长的加快和TNC (主要是淀粉) 含量的增加对氮的稀释造成的。Carbohydrates found in plants are frequently grouped into two different classes:structural carbohydrates and non-structural carbohydrates. The former mainlyconstruct the plant basic framework, while the latter are essential for plant growth andmetabolism. As yet there is lack of information on the effects of elevated CO2concentration on carbohydrate contents in stem, branch and root of plant, and oncarbohydrate allocation in organs of plant although there have been many reports onthe responses of carbohydrate contents to elevated CO2 concentration in plant foliages.A shift of carbohydrate contents in plant reflects a change in transporting ofphotosynthetic production from leaf to stem, branch and root of plant. The allocationof carbohydrates that is correlated to plant growth patterns affects plant growth. Thus,in order to understand the influences of elevated CO2 on biochemical process,physiological change and plant growth well, the response of carbohydrate contentsand allocation in plant to elevated CO2 should be further investigated. In our study, theeffects of elevated CO2 on carbohydrate contents and their allocation between leaf,stem, branch and root tissue of Betula albosinensis seedlings were determined. Theseedlings were grown in independent and enclosed-top chambers. Chambers werecontrolled to reproduce ambient (CK) and ambient + 350 µmol·mol-1 CO2 (EC)concentration for 1 year. The results here showed that,1) Elevated CO2 significantly increased non-structural carbohydrate contents in leafof red birch seedlings. This will reduce photosynthetic rate.2) Elevated CO2 also significantly increased non-structural carbohydrate contentsin root, stem and branch of red birch seedlings. These findings supported thehypothesis that elevated CO2 accelerated carbohydrates from leaf to branch, stem androot.3) Starch makes up the largest parts of total non-structural carbohydrate. In thesame way, the increase of starch plays a main role in the increase of totalnon-structural carbohydrate under elevated CO2. In leaf, branch, stem and root, theincrements of starch contents comprised 91.45%, 88.23%, 83.23% and 82.01% of theincrements of total non-structural carbohydrate contents.4) Under elevated CO2 the cellulose contents have an increasing tendency in redbirch seedlings. It is needed to investigate the effects of long-term elevated CO2 oncellulose contents in plant.5) There are significant CO2 effects on the allocation of carbohydrate in organs ofred birch seedlings. Under elevated CO2 more carbohydrates were allocated to root.Moreover, CO2 enrichment significantly increased the root to shoot ratio of red birchseedlings and the dry weight of roots. These results supported Gorissen and Cotrufo ‘shypothesis that increase of carbohydrate allocation to root mostly contributed to theincrease of root to shoot ratio.6) Elevated CO2 brought about a reduction in the nitrogen contents of leaf, stem,branch and root. The decline of nitrogen contents under elevated CO2 is mainlycaused by the dilution effects of increasing starch level and growth of red birchseedlings.

贡嘎山植物小枝功能性状在不同生活型和不同海拔间的比较研究

植物功能生态学研究不仅提供了植物生理生态学与生态系统生态学的连接，还为植物种群生活史对策研究提供了材料。Westoby 等 (2002) 提出了利用植物功能性状变量的主导维度来确定和量化植物生活史的生态适应策略。在他们所提出四个主导维度中，叶大小－小枝大小是研究相对较少的一维；其内部各组分的关系、对环境的响应，以及与其它重要维度的关系，目前的理解非常有限。 本研究以贡嘎山不同海拔不同功能群物种为研究对象，采用种间比较和系统发生独立性比较等研究方法，系统研究了植物的功能特征及其相关性在不同生境及不同功能群间的差异，旨在分析不同功能群物种的叶大小－小枝大小的成本和收益。其研究结果将有助于我们理解植物生活史对策的进化，进而理解物种共存和维持物种多样性的机制。主要研究结果如下： 1. 叶大小－小枝大小关系 小枝茎横截面积与单叶面积和总叶面积均呈异速生长关系，即总叶面积和单叶面积的增加比茎横截面积的增加速度快。但是，总叶面积和叶片干重的增加却基本上与小枝茎干重的增加等速。系统发生独立性比较研究的结果与此相一致。表明，在某一给定的茎投入时，至少大叶大枝物种不比小叶小枝物种在支撑叶面积和叶片干重方面具有优势。同时，在某一给定的小枝茎投入时，常绿阔叶物种比落叶阔叶物种支撑更少的叶面积。在茎干重与总叶面积的关系中，落叶复叶物种比落叶单叶物种具有更高的y轴截距，表明复叶物种比单叶物种在展叶面积方面更有效。复叶物种与单叶物种相比，通常具有较大的叶大小和小枝大小。 2. 叶大小－叶数量关系 叶大小与数量间在不同的叶片习性、不同的叶片形态以及不同的生境类型的物种间均存在稳定的负的等速生长关系，且这种关系在系统发生独立性比较时依然成立。然而，在某一给定的出叶强度 (单位小枝的叶数量) 时，常绿阔叶物种比落叶物种具有更小的叶面积。而在给定体积基础上的出叶强度时，落叶复叶物种的叶面积显著大于落叶单叶物种，且复叶物种比单叶物种具有更大的叶大小和更小的出叶强度。但是，叶大小与数量间的关系在不同的海拔间并没有显著的差异。 3. 小枝大小－总叶面积关系 在不同的生活型或不同的生境下，小枝上总叶面积与茎干重和小枝干重均呈正的异速生长关系，且斜率显著小于1.0，表明小枝上总叶面积的增加都不能赶上小枝及茎大小的增加。这种“收益递减”表明随着小枝干重的增加，光截取的收益递减。此外，叶面积比 (总叶面积与小枝干重的比值) 与单叶干重呈显著负相关关系，系统发生独立性比较的结果与此相一致。根据以上结果，可以推测，大叶的物种在质量较好的生境中出现，而群落内部小枝茎的寿命较长的物种可以拥有较大的叶片。 4. 叶片色素浓度－LMA关系 随着海拔的升高，阔叶木本植物和草本植物的叶片色素浓度减少，叶绿素a/b和类胡萝卜素/叶绿素比值以及比叶重 (LMA) 增加。然而，在草本植物中的色素浓度、色素比值和LMA的变化比阔叶木本植物的更明显。同时，LMA与叶片色素浓度呈负相关关系，但是在落叶物种中的LMA对色素浓度的影响比常绿阔叶物种更强烈。总之，草本植物的叶片特征对海拔梯度的变化似乎比木本植物更敏感，LMA对叶片色素的保护作用在落叶物种中比在常绿阔叶物种显得更重要。这些结果表明不同生活型物种可能采取不同的保护机制来降低叶绿体器官的损伤和增加他们的碳获取能力。 Studies on plant functional ecology not only bridge plant eco-physiology and ecosystem functioning, but also enrich plant population biology. As pointed out by Westoby et al (2002), plant life history strategies can be identified and quantified by four leading dimensions of variations in plant functional traits, i.e., seed size/output, leaf mass per area and leaf life span, plant height, and leaf size-twig size. Compared to the other dimensions, the cost/benefit of the leaf size-twig size spectrum has scarcely been analyzed in relation to environmental gradients and life form types, and the adaptive significance of this spectrum is not fully understood. In the present study, the relationships between functional traits of plant twigs are determined for the species with different life forms along an altitudinal gradient of Gongga Mountain with both cross-species analysis and evolutionary divergence analysis. The primary objective of this study is to examine the cost/benefit of leaf size-twig size in plants. The study results are supposed to provide insights into the understanding of the mechanism of species coexistences. The results are shown in the following. 1. The relationship between leaf size and twig size Twig cross-sectional area allometrically scaled with both individual leaf area and total leaf area supported by the twigs. However, the increase in total lamina mass/area was generally proportional to the increase in stem mass. These correlations between trait variations were significant in both interspecies analysis and phylogenetically independent comparison (PIC) analysis, which indicated that thick-twigged/large-leaved species, at least, do not have an advantage in supporting leaf/lamina area and lamina mass for the same twig stem investment than thin-twigged/ small-leaved species. Meanwhile, the evergreen broad-leaved species supported a smaller leaf area for the same twig stem investment in terms of both cross-sectional area and stem mass than the deciduous species. The deciduous compound-leaved species have a higher y-intercept in the scaling relationship of twig stem mass versus total leaf area than the deciduous simple-leaved species, indicating that compound-leaved species were more efficient in displaying leaf area. The compound-leaved species were larger in both leaf size and twig size than their counterpart in the present study. 2. The relationship between leaf size and leaf number Significantly negative and isometric scaling relationships between leaf size and leafing intensity (leaf number per twig mass or volume) were found to be consistently conserved across species independent of leaf habit, leaf form and habitat type. The negative correlations between leaf size and leafing intensity were also observed across correlated evolutionary divergences. However, leaf area was smaller in the evergreen broad-leaved species at a given leafing intensity than in the deciduous species. The deciduous compound-leaved deciduous species were higher in leaf area than deciduous simple-laved species at a given volume-based leafing intensity. Moreover, the compound-leaved deciduous species were larger in leaf size but smaller in leafing intensity than their simple counterparts. No significant difference was found in the scaling relationships between altitudes. 3. The relationship between twig size and total leaf area Leaf area was found to scale positively and allometrically with both stem and twig mass (stem mass plus leaf mass) with slopes significantly smaller than 1.0, independent of life form and habitat type, indicating that the increase in total leaf area fails to keep pace with increasing twig size and stem size. This ‘diminishing returns’ suggests that the benefit of light intercept decreased with increasing twig mass. Moreover, the leaf area ratio (the ratio of total leaf area to stem or twig mass) correlated negatively with individual leaf mass. The results of PIC were consistent with the correlations. According to the results, it is speculated that large-leaved species may be favored when habitat is good and when stem longevity are long within community. 4. The relationship between leaf pigment concentrations and leaf mass per area With increasing altitude, the concentrations of pigments decreased, but the ratios of chlorophyll a/b and carotenoid/chlorophyll, and LMA increased, in both the broad-leaved woody species and herbaceous species groups. However, the changes in the pigment concentrations, ratios and LMA were more profound in the herbaceous species than in the woody species. In addition, pigment concentrations were negatively correlated with LMA in each life form type and in the pooled dataset. However, the LMA effect on leaf pigment concentrations was more profound in the deciduous species than in the evergreen braode-leaved species. In general, herbaceous species seemed more sensitive to the increasing altitude compared to woody species, and LMA seemed to be a more important mechanism for protecting leaf pigments in deciduous species than in evergreen broad-leaved species. These results suggested that the species with different life forms may employ different protective mechanisms to decrease the chloroplast apparatus damage and increase their carbon gain.

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.

坡面植物生长期内对~7Be的截留吸收及影响因素分析

对坡面植物在生长期内7Be含量的动态变化进行研究,发现随着生长植物在生长期内7Be含量增加,单位面积上植物截留吸收7Be量增加;研究区坡面7Be的植被截留吸收率为3.35%~8.44%,说明植被覆盖对土壤中7Be含量有重要的影响。同时还发现,不同采样间隔期7Be日均增量不定,在整个采样期内变异性大,变异系数为0.69,但从春季到夏季的前4个采样间隔期变异性相对较小;不同采样间隔期7Be日均增量和日均降雨量呈正比,相关系数为0.64。

植物化感作用与生物多样性

本文简要地阐释了化感作用的含义、基本特征以及作用机制,并结合生物多样性理论,综述了化感作用研究中化感物种的多样性、化感物质的多样性及其释放途径的多样性,具体讨论了化感作用对物种多样性、遗传多样性及生态系统多样性中的种群生态、协同进化、土壤生境、生态系统功能和生物入侵等方面的可能影响。文中提出了化感作用的利用、管理应与生物多样性保护相统一的看法,并指出对化感作用与生物多样性的关系以及相互影响机制进行本质的探索,特别是对植物化感作用的生态服务功能与价值评估与探讨,可为保护生物学和系统生态学提供理论基础,这也是今后工作开展的一个重要方向。

Comparative Study on Antioxidant Systems of Polygonum Viviparum Grown at Two Different Altitudes

Study on the antioxidant systems of Polygonum viviparumgrown at two different altitudes indicated plants grown at Haibei Research Station at 3200 m altitude as compared with plants grown in Xining at 2300 m altitude had apparently higher contents of ultroviolet-absorbing compounds and ascorbic acid, and significantly higher activities of superoxide dismutase, peroxidase and ascorbic peroxidase. Higher contents of superoxide radical anions and malonadehyde were also found in plants at Haibei Research Station as compared with the plants grown in Xining which have been transplanted from Haibei Research Station for at least four years. The differences in antioxidant system reflect a long term of time of adaptation to different environments.

Variation of active constituents of an important Tibet folk medicine Swertia mussotii Franch. (Gentianaceae) between artificially cultivated and naturally distributed

Concentrations of seven phytochemical constituents (swertiamarin, mangiferin, swertisin, oleanolic acid, 1,5,8-trihydroxy-3methoxyxanthone, 1,8-dihydroxy-3,7-dimethoxyxanthone and 1,8-dihydroxy-3,5-dimethoxyxanthone) of "ZangYinChen" (Swertia mussotii, a herb used in Tibetan folk medicine) were determined and compared in plants collected from naturally distributed high-altitude populations and counterparts that had been artificially cultivated at low altitudes. Levels of mangiferin, the most abundant active compound in this herb, were significantly lower in cultivated samples and showed a negative correlation with altitude. The other constituents were neither positively nor negatively correlated with cultivation at low altitude. Concentrations of all of the constituents varied substantially with growth stage and were highest at the bud stage in the cultivars, but there were no distinct differences between flowering and fruiting stages in this respect. (c) 2005 Elsevier Ireland Ltd. All rights reserved.