芦苇Phragmites communis Trin. 耐盐变异体的生物学鉴定和快速繁殖以及在海水生态养殖中的应用示范

本文以芦苇野生亲本和耐盐变异体为材料，比较了两者在形态、生理生化以及分子生物学特性上的差异，对耐盐变异体抗盐能力提高的机理作了初步的探讨。为了适应推广芦苇耐盐变异体的需要，进行了耐盐变异体快速繁殖的研究，建立了两种诱导丛生芽的技术系统。对耐盐变异体芦苇在滩涂上的利用作了有益的尝试。结果总结如下： 1．通过对芦苇野生亲本和耐盐变异体的基因组DNA用随机引物扩增分析，发现两者的基因组DNA在序列上存在着一定的差异，并克隆测序了几个对变异体而言是特异的标记序列。 2．生化分析发现，在盐胁迫下，两者在可溶性蛋白上存在着差异，芦苇耐盐变异体在胁迫下分别在20～30 kD和43～66．2 kD之间各有一条特异的蛋白带表达。而且变异体盐胁迫下在同工酶的表达上也与野生亲本有着显著的区别。 3．生理测定发现，芦苇耐盐变异体在200 mmol/L NaCl盐胁迫下光合作用要比野生亲本强，叶绿素测定的结果与此相吻合，在此浓度的胁迫下，变异体叶绿素含量受影响较小，而野生亲本的叶绿素含量明显降低。对两者胁迫前后的离子含量测定发现，虽然K+含量最多，但是植株内离子含量变化最大且增加最多的离子是Na+，而且变异体内Na~+增加的量比野生亲本高得多。另一个变化较大的是游离脯氨酸的含量，其变化情况类似于Na~+，在胁迫后脯氨酸含量增加明显，而且变异体内的增加量比野生亲本高。对变异体进一步的胁迫反应证实了Na+和脯氨酸含量变化与胁迫反应的密切联系。推测它们的这些变化与变异体抗盐能力提高密切相关。 4．将芦苇耐盐变异体的种子苗切去种壳和种子根，以此作为外植体，通过筛选大量的激素组合，最终建立了两种快繁技术系统（MP－A和MP－B）进行丛生芽诱导。两个系统都包含预处理和诱导两个主要步骤，其中预处理培养基激素组合是相同的（NAA 2～5 mg/L + 2,4-D 0.05 ～ 0.1 mg + BA 1mg/L）,而诱导处理的培养基激素组合不同，分别为：MP－A的诱导处理的激素组合是将预处理的激素组合中的2，4－D去除即可;MP－B诱导处理的激素组合为1mg/L NAA + 0.05 ～0.1mg/L 2,4-D + 2～5mg/L BA. 两个系统都获得显著的丛生芽诱导效果。 5．在滩涂水产养殖中引入芦苇种植、发现耐盐芦苇能有效地降低水体污染和病害，实验了两种模式的种养殖方式：围隔模式和混合模式，初步结果表明混合模式效果较好。这一初中初步验证了芦苇对海水养殖的价值，对开发滩涂具有很大的意义，具有创新性。

I.小麦非叶器官碳酸酐酶活性与定位分析II.镉毒害导致芦苇胚性细胞死亡的激素调控研究

第一部分： 通过生理测定和化学染色分析了冬小麦品种小堰54和京411的叶片和非叶片组织的碳酸酐 酶活性。叶片碳酸酐酶活性(CA)在挑旗时期达到最大值，之后减少到最小，而在饱粒期又呈 增加趋势。从灌浆期到饱粒期，颖片和内稃的CA活性均减少，而外稃和种皮的CA活性均增加。在饱粒期，小堰54的叶片、颖片、外稃和种皮CA活性均高于京411。组织化学染色表明，CA主要分布在旗叶的叶肉细胞叶绿体中，也分布在非叶片组织颖片、外稃和内稃的叶肉和维管束鞘细胞的胞质中。这些结果表明，小麦非叶组织叶肉和维管束鞘细胞的胞质中的CA可能对饱粒期冬小麦的C4光合途径起作用。饱粒期小堰54的C02传递到Rubisco酶速率和抗旱性较京411高。 第二部分： 以继代培养的芦苇胚性细胞为材料，利用台盼兰拒染法检测了悬浮细胞死亡过程，并利用石蜡切片法及苏木精染色法观察了不同浓度镉对芦苇细胞的毒害作用。1000μM的CdCl2迅速导致芦苇悬浮细胞死亡，200μM的CdClz在接种后第5天引起悬浮细胞死亡，100μM的CdCl2在接种后第7天引起悬浮细胞死亡，≤50μM的CdClz在接种后7天不引起悬浮细胞死亡。同时对不同浓度镉处理的芦苇胚性细胞的内源植物激素和可溶性蛋白质进行分析，≤50μM的镉浓度显著地降低胚性细胞内IAA、ZR、GA3和GA4的含量，却提高ABA的含量，抑制可溶性蛋白质的合成：≥100μM的镉浓度显著地提高IAA、ZR、GA3和GA4的含量，却降低ABA的含量，促进可溶性蛋白质的合成。这些结果表明，镉的毒害至少包括镉浓度决定的两种细胞死亡机制，高浓度的镉(1000μM)引起的细胞死亡应当为坏死，而100μM的镉引起植物悬浮细胞发生程序性死亡。在较高浓度(≥100μM)的镉处理下，芦苇细胞内内源IAA、ZR、GA3和GA4的浓度较高，可能调控可溶性蛋白质的合成而促进细胞发生程序化死亡。

人工湿地强化净化太湖水体富营养化研究

为了进一步降低水中营养盐浓度、提高水体透明度，为湖泊生态系统的恢复创造条件，促进湖泊水体生态系统良性循环，以太湖五里湖富营养化水体为研究对象，采用中试规模的人工湿地技术来强化净化富营养化水体。重点研究了不同流态、不同基质、不同植物及植物有无条件下各人工湿地系统对富营养化水体的处理效果。此外，还对生长在人工湿地中的7种湿地植物芦苇(Phragmites communis)、香蒲(Typha angustifolia)、茭白(Zizania caduciflora)、水葱(Scirpus validus.)、菖蒲(Acorus calamus)、鸢尾(Iris pseudacorus)和千屈菜(Lythrum salicaria)的生长、生理生态特性如气体交换、叶绿素荧光、光合色素含量及氮、磷吸收能力进行了研究，旨在探索高等植物在富营养化水体净化过程中的机理和规律，为人工湿地污水净化机理奠定一定的科学和理论基础。主要研究结果如下： 通过对垂直潜流、水平潜流和自由表面流三种类型的人工湿地系统对富营养化水体净化效果的对比研究，我们发现在水力负荷0.64 m d-1，进水污染物浓度化学需氧量(COD) 7.37 mg L-1、氨氮(NH4+-N) 1.63 mg L-1、硝氮(NO3--N )1.41 mg L-1、总氮(TN) 4.82 mg L-1和总磷(TP) 0.15 mg L-1的条件下，三种不同流态的人工湿地对富营养化水体均有一定的净化功能。三种类型的人工湿地对主要污染物COD、NH4+-N、NO3--N、TN和TP的去除效果分别为：垂直潜流40.4%、45.9%、62.9%、51.6%和64.3%;水平潜流39.6%、32.0%、65.3%、52.1%和65.7%;自由表面流16.5%、22.8%、34.2%、19.8%和35.1%。相比之下，垂直潜流、水平潜流对主要的污染物的去除效果明显好于自由表面流人工湿地。除垂直潜流湿地NH4+-N的平均去除率显著大于水平潜流外，两人工湿地在其余指标的去除效果上无显著差异。考虑到土地面积的限制，就太湖五里湖富营养化水体治理而言，垂直潜流和水平潜流较自由表面流更为适合。 以沸石为基质的人工湿地较以砾石为基质的人工湿地对NH4+-N有很好的去除效果，除此之外两种基质类型的人工湿地在其余各污染指标的去除效果方面差异不显著。但随着湿地运行时间的延长，沸石对NH4+-N的吸收能力会逐渐减弱，考虑到湿地构建成本，建议就地取材，以当地较为廉价的砾石为人工湿地基质比较经济适用。 有无植物对有机污染物COD的去除效果影响差异不显著，但对TN和TP去除效果影响差异极显著，有植物的人工湿地对TN的去除率比无植物的分别高出12.5%和13.4%，对TP去除率分别高出16.9%和31.0%。不同植物对人工湿地处理效果的影响差异不显著，可见湿地流态对处理效果的影响大于植物的影响。 各湿地植物叶片的净光合速率(Pn)日变化均为双峰曲线，有光合“午休”现象。其中芦苇、茭白、鸢尾、菖蒲、水葱和千屈菜Pn的下降主要是由气孔导度(gs)的下降造成的，而香蒲则更多的受叶肉光合能力下降的影响。除千屈菜的Pn与gs之间呈显著正相关外，其余植物的Pn与gs之间均呈极显著正相关。气孔行为对湿地植物光合作用碳的固定显示了明显的主导控制作用。 各湿地植物的光补偿点(LCP)差异不显著，其均值都在10µmol m-2 s-1以上，显示了阳生植物的特性。但湿地植物的光饱和点 (LSP)差异极显著，LSP大小依次为香蒲1476.3µmol m-2 s-1>水葱1140.0µmol m-2 s-1>菖蒲753.7µmol m-2 s-1>芦苇751.7µmol m-2 s-1>茭白640.7µmol m-2 s-1>千屈菜567.3µmol m-2 s-1>鸢尾479.0µmol m-2 s-1。7种湿地植物的CO2补偿点差异极显著，CO2补偿点大小依次为水葱47.1µmol mol-1>茭白28.4µmol mol-1>香蒲23.7µmol mol-1>鸢尾16.8µmol mol-1>菖蒲16.7µmol mol-1>千屈菜15.2µmol mol-1>芦苇14.8µmol mol-1。与此相反，湿地植物的CO2饱和点差异不显著。 各湿地植物的Fv/Fm值大小差异也极显著，Fv/Fm值大小依次为千屈菜0.8168>水葱0.8348>香蒲0.8262>菖蒲0.8198>芦苇0.8168>茭白0.8040>鸢尾0.7930。由此可见千屈菜、水葱较芦苇和茭白有较高的PSⅡ效率。7种湿地植物叶片的叶绿素a、叶绿素b和类胡萝卜素含量水平差异极显著，芦苇和茭白叶片的叶绿素和类胡萝卜素含量水平都较高，这在客观上解释了他们为何拥有较强的光合能力。 不同湿地植物的生物量大小差异极显著，平均生物量(干重)变化范围在0.26-6.65 kg m-2之间。除香蒲和水葱的地下部生物量显著大于地上部外，其余植物都是地上部生物量大于地下部，这非常有利于人工湿地生态工程中通过收获植物地上部生物量来达到去除氮、磷污染目的。 不同湿地植物植株氮、磷含量差异极显著。湿地植物对氮吸收总量变化范围为6.09-93.96 g m-2，吸收氮最高是芦苇，最低是菖蒲;对磷吸收总量变化范围为0.51-8.95 g m-2，吸收磷最高是香蒲，最低是菖蒲。经测算，植物吸收的总氮量占人工湿地总氮去除量的0.6-17.1%，总磷量占人工湿地总磷去除量的1.4-40.5%。本次试验中香蒲和芦苇对氮、磷的吸收去除能力比较高，茭白、鸢尾、水葱和千屈菜的吸收去除能力中等，而菖蒲的吸收能力较弱。

Fluorescence-polarization studies on binding of 4-methylumbelliferyl beta-D-galactopyranoside to Ricinus communis (castor-bean) agglutinin

The binding of Ricinus communis (castor-bean) agglutinin 1 to saccharides was studied by equilibrium dialysis and fluorescence polarization by using the fluorescently labelled sugar 4-methylumbelliferyl beta-D-galactopyranoside. No appreciable change in ligand fluorescence of 4-methylumbelliferyl beta-D-galactopyranoside was considerably polarized on its binding to the lectin. The association constants obtained by Scatchard analysis of equilibrium-dialysis and fluorescence-polarization data do not differ much from each other, and at 25 degrees C, Ka = 2.4 (+/- 0.2) X 10(4)M-1. These values agree reasonably well with that reported in the literature for Ricinus agglutinin 1. The number of binding sites obtained by the different experimental procedures is 1.94 +/- 0.1 per molecule of 120 000 daltons and is equal to the reported value of 2. The consistency in the values of Ka and number of binding sites indicate the absence of additional subsites on Ricinus agglutinin 1 for its specific sugars. In addition, the excellent agreement between the binding parameters obtained by equilibrium dialysis and fluorescence polarization indicate the potential of ligand-fluorescence-polarization measurements in the investigation of lectin-sugar interactions.

Chemical Modification Studies On Ricinus-Communis (Castor Bean) Agglutinin

Ricinus communis agglutinin was subjected to various chemical treatments and the effect on its hemagglutinating and saccharide-binding properties was studied. Acetylation, succinylation and citraconylation led to a complete loss in the activity of the agglutinin, whereas reductive methylation had no effect on the activity, showing that charged amino groups were involved in the hemagglutinating and saccharide-binding activity of Ricinus agglutinin. Modification of tryptophyl, arginyl and carboxyl-group-containing residues did not lead to any loss in the activity of the agglutinin. Acetylation of tyrosyl groups with N-acetylimidazole strongly reduced the hemagglutinating and saccharide-binding property of Ricinus agglutinin. The loss in activity was restored on deacetylation of the tyrosyl groups. Modification of tyrosyl residues also led to a change in the immunological properties of the agglutinin. The initial rate of modification of tyrosyl and amino groups and the concomitant loss of activity was reduced in the presence of lactose.

On the Specificity of Carbohydrate-Lectin Recognition The Interaction of a Lectin from Ricinus communis Beans with Simple Saccharides and Concanavalin A

A galactose-specific protein (RC1) isolated from Ricinus communis beans was found to give a precipitin reaction with concanavalin A. Its carbohydrate content amounted to 8–9% of the total protein and was found to be rich in mannose. The interaction of RC1 with galactose and lactose was measured in 0.05 M phosphate buffer containing 0.2 M NaCl (pH 6.8) by the method of conventional equilibrium dialysis. From the analysis of the binding data according to Scatchard method the association constant (Ka) at 5°C was calculated as 3.8 mM−1 and 1.2 mM−1 for lactose and galactose, respectively. In both cases the number of binding sites per molecule of RC1 with molecular weight of 120000 was found to be 2. From the temperature-dependent Ka values for the binding of lactose, the values of –5.7 kcal/mol and –4.3 cal × mol−1× K−1 were calculated for ΔH and ΔS, respectively. The addition of concanavalin A to RC1 or vice versa led to the formation of the insoluble complex RC1· ConA4 containing one molecule of RC1 and one molecule of tetrameric concanavalin A (ConA4) which could be dissociated upon addition of concanavalin A-specific sugars. The complex formation results in a time-dependent appearance of turbidity in the time range from 10s to 10 min. From the measurement of the time-dependent appearance and disappearance of the turbidity the formation (kf) and dissociation (kd) rate constants were calculated as 3 mM−1× s−1 and 0.07 ks−1 respectively. The ratio kf/kd (43μM −1), that corresponds to the association constant of complex RC1· ConA4, is higher than that of mannoside · ConA4 and thereby suggests that protein-protein interaction contributes significantly in stabilising glycoprotein · lectin complexes. The relevance of this finding to the understanding of the chemical specificities that are involved in a model cell-lectin interaction is discussed.

Dynamics of carbohydrateâlectin interaction The interaction p-nitrophenyl-β-D-galactose with a lectin from Ricinius communis

L&in-induced agglutination is a complex process determined by several factprs such as the nature of lectin (valency, binding constant) the properties of cell membrane (fluidity, distribution of lectin receptor sites) and the metabolic state of the cell (microvilli, microtubules, microfilament) [l-3].

Res Communis? A critical legal geography of outer space, Antarctica, and the deep seabed

This chapter provides a critical legal geography of outer Space, charting the topography of the debates and struggles around its definition, management, and possession. As the emerging field of critical legal geography demonstrates, law is not a neutral organiser of space, but is instead a powerful cultural technology of spatial production. Drawing on legal documents such as the Outer Space Treaty and the Moon Treaty, as well as on the analogous and precedent-setting legal geographies of Antarctica and the deep seabed, the chapter addresses key questions about the legal geography of outer Space, questions which are of growing importance as Space’s available satellite spaces in the geostationary orbit diminish, Space weapons and mining become increasingly viable, Space colonisation and tourism emerge, and questions about Space’s legal status grow in intensity. Who owns outer Space? Who, and whose rules, govern what may or may not (literally) take place there? Is the geostationary orbit the sovereign property of the equatorial states it supertends, as these states argued in the 1970s? Or is it a part of the res communis, or common property of humanity, which currently legally characterises outer Space? Does Space belong to no one, or to everyone? As challenges to the existing legal spatiality of outer Space emerge from spacefaring states, companies, and non-spacefaring states, it is particularly critical that the current spatiality of Space is understood and considered.

Manual Control of Phragmites australis on Pondshores of Cape Cod National Seashore, Masachusetts, USA

The non-native, invasive genotype of the common reed ( Phragmites australis (Cav.) Trin. ex Steudel) has become a problem of significant proportions throughout wetlands of North America (Saltonstall 2001). Although attempts to suppress or eradicate Phragmites have utilized a wide variety of techniques, herbicides have generally been most effective (Marks et al. 1994). In the spring, mid-summer, and late summer of 2003, we attempted to opportunistically control Phragmites in five freshwater ponds within Cape Cod National Seashore (CCNS) by repeatedly severing stems underwater, at ground level.(PDF has 4 pages.)