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.

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.

干旱胁迫下沙地樟子松脱落酸变化及生理响应

土壤水分是限制沙地樟子松生长发育的关键因子。采用根系渗透胁迫法（ＰＥＧ处理）和盆栽干旱处理。以幼苗根系及叶片中脱落酸（ＡＢＡ）含量的累积作为幼苗对干旱胁迫响应的指示剂．模拟沙地樟子松幼苗对干旱胁迫的生理生态响应。在干旱胁迫下．根部和叶片ＡＢＡ均可迅速积累．但叶片ＡＢＡ累积较根部滞后；同时测定干旱胁迫下气孔导度、蒸腾速率、净光合速率．认为ＡＢＡ增加可降低气孔导度、并伴随蒸腾速率、净光合速率下降。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。

海洋光合细菌Rx菌株的分离鉴定与生物学特性研究

　从大连金州区海湾养虾池底分离获得光合细菌Rx菌株的初筛菌株,并对其纯培养物进行了形态特征、培养特征、生理生化特征以及DNA中G+C摩尔百分比等生物学特性分析。结果表明,Rx菌株为革兰氏阴性菌,大小为(0.5～0.7)μm×(1.0～1.2)μm,单极生鞭毛,光合内膜为泡囊状,繁殖方式为二分分裂生殖;Rx菌株的纯培养物含细菌叶绿素a和球状素型类胡萝卜素,光照厌氧和黑暗好氧条件均能生长;Rx菌株生长盐度为5～100g/kg,生长pH为5.8～8.4;Rx菌株不能利用硫化物为电子供体,其生长受高浓度硫化物的抑制;能利用谷氨酸而不能利用柠檬酸钠、酒石酸为有机碳源或电子供体;盐酸硫胺素(t)、生物素(b)、p-对氨基苯甲酸(p-ABA)、烟酸(n)为Rx菌株生长的必需因子;Rx菌株DNA的G+C摩尔百分比为64.18%。根据以上特征,并参照《常见细菌系统鉴定手册》(2002)和《伯杰氏细菌系统学手册》第3卷(1989),将Rx菌株鉴定为小红卵菌属广海小红卵菌(Rhodovulumeuryhalinum)。

根系吸水机理研究进展

植物根系除支撑和固定其地上部这一力学功能外的一个重要功能就是从土壤中吸收水分和养分以满足植物地上部生长所需.表征植物根吸水能力的一个重要的水力学参数是水力导度(用单位时间单位面积的水流速率来表示),可在细胞(细胞水力导度)、单根和整株根系水平上来表达,其中单根导度可分为径向导度和轴向导度,仅径向导度反映了单根吸收水分的能力,而轴向导度则反映了植物根系输导水分的能力,但在整株根系水平上则以通过整个根系的水流通量与根木质部和根表土壤间的水势差之比来表示,既包括径向导度也包括轴向导度.近年来,对植物根系吸水或根水力导度研究已取得了许多重要进展,这对阐明根系吸水机理和地上地下部关系起到了重要作用.下面做一简要综述.1　根系吸收水分的主要部位根系吸水的部位一般认为在距根尖10～100ｍｍ的区域内,这种看法是基于解剖上的证据.从根表面到根中心,依次为根表皮、下表皮、皮层、内皮层、中柱,其中根表皮是有最高吸收活性的根区,但一般仅可存活几天,而内皮层将皮层和中柱分开形成了根内侧的一个界面,老根一般有周皮或栓质化的内皮层,有很强的不透水性(即阻力很大).但Ｓａｎｄｅｒｓｏｎ[1]对大麦的研究发现,虽然老根区内重度栓化的内皮层已...

内源激素含量的变化与白桦成花关系的研究

采用酶联免疫吸附法(ELISA)测定有花史白桦大树与无花史白桦幼树的4种内源激素脱落酸(ABA)、细胞分裂素(iPA)、生长素(IAA)、赤霉素(GA3)含量的动态变化,研究其与白桦成花的关系。研究结果表明:白桦有花史大树与无花史幼树的4种内源激素含量动态变化明显不同。在雌、雄花芽的生理发端期,有花史大树的IAA含量保持相对稳定,而高含量的ABA和iPA含量的逐渐降低以及GA3含量的逐渐升高可能有利于白桦成花。

干旱下植物气孔运动的调控

概述了植物气孔对大气干旱和土壤干旱的反应 ,认为植物气孔对大气干旱的反应并不是一种反馈机制 ;并就干旱条件下植物气孔运动的水力学和化学信号调控机制进行了简要论述 ,认为虽然化学信号调控干旱下气孔运动更为广泛 ,但 ABA不是唯一的化学信号 ,水分关系影响了信号的产生、运转和气孔对信号的敏感性 ,干旱条件下水力学和化学信号共同调控着植物的气孔运动

平阴玫瑰花芽分化期叶片内源激素的变化

对平阴玫瑰花芽分化期叶片甲醇提取物进行IAA、ZR、GA3、ABA的分离、纯化和测定。结果发现,所测的几种激素均表现出明显的变化规律,其中IAA和GA3在花芽分化期含量逐渐下降,且在分化临界期出现一低峰,而ZR和ABA则完全相反。同时经比较分析得出ABA/GA3,ABA/IAA,ZR/GA3,ZR/IAA也表现明显的变化规律,即比值总体趋势是逐渐提高,且均在分化临界期含量出现一飞跃,显然ABA/GA3,ABA/IAA,ZR/GA3,ZR/IAA在平阴玫瑰的花芽分化过程中起着重要的调控作用,由此推测,增加植物体内的ABA、ZR的含量或降低IAA、GA3的含量,都可以促进玫瑰的花芽分化;反之则抑制其花芽分化。

干旱对沙棘休眠、萌芽期内源激素及萌芽特性的影响

以 2a生中国沙棘为试验材料 ,研究了土壤干旱胁迫下沙棘苗木水分生理状况 ,内源GA1 3 及ABA水平与萌芽率的关系 ,结果表明 :土壤干旱胁迫导致苗体含水量、水势、自由水含量下降 ,显著提高冬季休眠与春季萌动期的内源ABA水平而降低内源GA1 3 含量 ,GA1 3 ABA降低 ,较晚达到萌动所需的调控阈值 ,萌芽延迟。重度胁迫下苗木延迟达 2 5d且萌芽后生长缓慢。长期土壤中度干旱条件下沙棘苗萌芽期亦略有延迟 ,萌芽后生长略有抑制

干旱胁迫对植物基因的诱导及基因产物的变化

对干旱胁迫所诱导的基因及其产物的功能进行了综述。干旱基因分为 3种类型 :ABA依赖型、ABA非依赖型和不能被 ABA诱导型。这些基因编码的蛋白质在植物适应干旱的过程中发挥着重要的作用。这对揭示植物抗旱机制具有重要意义

土壤镉污染毒性效应的多指标综合评价

以草甸棕壤为供试土壤,以蚕豆幼苗根尖有丝分裂指数、染色体畸变率以及微核率,幼苗叶片内抗氧化酶活性、植物内源激素含量为指标,采用盆栽方法研究了0~10 mg.kg-1镉胁迫对植物细胞的遗传和生态毒性效应.结果表明,在此浓度范围内,蚕豆根尖细胞有丝分裂指数、染色体畸变率以及微核率均随镉浓度增加呈显著的剂量-效应正相关关系,其中微核率变化最为明显,处理组微核率分别为对照组的1.43~3.22倍;蚕豆幼苗叶片内的SOD和POD活性变化呈先升高后下降的趋势;而CAT活性随镉浓度增加其变化规律与SOD、POD相反.此外,镉胁迫下植物激素脱落酸(ABA)、赤霉素(GA3)与细胞分裂素类的玉米素和玉米素核苷总含量(Z&ZR)均表现出低浓度下诱导和较高浓度下诱导率降低的趋势,在镉浓度为2.5 mg.kg-1时3种植物激素含量均达到最高值,分别比对照组增加了6.6%、4.0%和12.6%.研究表明,各指标对污染物的毒性具有响应且响应的域值及其敏感度不同,将各指标综合使用可使土壤镉污染的遗传和生态毒性诊断更为有效.

脱落酸对UV-C胁迫下小麦幼苗光合特性及抗氧化酶活性的影响

研究了UV-C辐射下短期和长期脱落酸(ABA)处理对小麦幼苗CO2同化作用、羧化效率、光合CO2响应以及抗氧化酶活性等的影响.结果表明,在无UV-C辐射情况下,短期和长期ABA处理能提高光合速率,比对照增加14·69%和20·46%,降低气孔导度,比对照降低14·74%和17·31%,但对胞间CO2浓度和羧化效率影响不大.当受到UV-C辐射时,光合速率、羧化效率、气孔导度和胞间CO2浓度逐渐降低.长期ABA处理变化最小,其次为ABA短期处理,对照降低最大.ABA处理能够提高小麦光合对CO2的响应,UV-C辐射抑制光合对CO2的响应.ABA处理能够提高小麦抗氧化酶(CAT、SOD、POD)活性而降低MDA含量.在UV-C辐射下,CAT活性先升高随后降低,在辐射处理1h时活性达最大值,ABA处理的SOD和POD活性先升高后降低,且ABA长期处理比短期处理增加明显,对照则逐渐降低.ABA处理可能通过提高小麦CO2同化作用和抗氧化酶活性增强对UV-C胁迫的抗性,且ABA长期处理比短期处理效果更明显.

脱落酸对小麦幼苗光合及抗氧化酶的影响

研究了短期和长期外源脱落酸(ABA)处理对小麦幼苗CO2同化作用、羧化效率、光合CO2响应以及抗氧化酶等的影响.外源ABA处理能提高小麦叶片光合速率,但使气孔导度、胞间CO2浓度和羧化效率略有降低.ABA处理能够提高小麦光合对CO2的响应.无论是长期还是短期ABA处理都能够不同程度提高小麦抗氧化酶(CAT、SOD、POD)活性,且对SOD影响最明显.结果表明,ABA处理可能主要通过提高小麦抗氧化酶活性从而增强对环境胁迫的抗性,且长期处理效果好于短期处理.

一氧化氮是脱落酸诱导杨树叶片气孔关闭的信号分子

研究了外源NO和ABA对杨树气孔运动调节作用 .结果表明 ,外源NO和ABA都能诱导杨树离体叶片气孔关闭 ,且具有剂量效应 ,NO可加强ABA诱导气孔关闭的作用 .NO清除剂 (c PTIO)可大大减弱NO和ABA对气孔关闭的诱导作用 .证实了NO参与ABA调控气孔开闭运动过程 ,不同浓度NO供体SNP和ABA处理杨树离体叶片 ,SOD活性变化不明显 ,POD活性受到显著抑制 .杨树叶片粗酶液的体外实验表明 ,不同浓度SNP对POD活性的抑制呈明显的浓度及时间效应 ;而ABA对POD活性则几乎没有影响 .本研究证明 ,NO调节ABA诱导的树木气孔关闭作用 ,是ABA诱导树木气孔关闭的一种重要信号分子 .

玉米抗旱性评价指标的系统分析

对可用于玉米抗旱性评价的指标进行了系统研究,并借引了灰色关联度分析法进行了评价指标的筛选。研究结果表明:对于一个玉米品种进行抗旱性鉴定,不仅需进行多指标的重复测定,且需综合评定各生育时期的抗旱性。因而提出种子萌发胁迫指数、干旱后幼苗存活率、抗旱系数三位一体的初级鉴定指标,同时结合光合速率、ABA含量、NRase活性等次级鉴定指标的玉米抗旱性鉴定指标体系。表1,参28。