利用反义RNA技术进行木质素生物合成调控的研究

木质素是一类酚类次生代谢产物，在植物体内行使重要的生理功能，但它却是形成造纸污染的主要来源。利用基因工程手段，在分子水平调节木质素的生物合成，降低木质素的含量或改变组分以培育适合造纸的植物原料树种具有较大的应用价值和环保效益。本研究利用反义RNA技术，主要围绕木质素合成三种相关酶咖啡酸-O-甲基转移酶（COMT）、咖啡酰辅酶A-O-甲基转移酶(CCoAOMT)、4-香豆酸：辅酶A连接酶（4CL）的基因对植物木质素生物合成途径调节的研究，取得如下进展： 1.农杆菌介导法将COMT和CCoAOMT基因的单价和双价的反义表达载体导入烟草，比较了两个甲基化酶的功能。PCR-Southern和Northern点杂交结果表明反义基因已整合到烟草基因组DNA上，并在转录水平表达。两种反义基因对木质素生物合成调节的效果显示，CCoAOMT能更有效地调节木质素生物总量的合成，COMT仅特异调节S木质素的合成。表达反义CCoAOMT基因的转基因毛白杨，内源CCoAOMT基因的表达在转录和蛋白水平均受到抑制，最终引起转基因植株木质素含量普遍降低，最多降低达26.20%，筛选出木质素含量下降10%以上的转基因毛白杨株系8个，为源头治理造纸废水污染奠定了基础。 2． 对克隆的4CL基因进行了表达特性分析， RT-PCR分析表明，分离的毛白杨4CL基因主要在木质部丰富表达，叶中表达量较少，树皮中不表达。在毛白杨的一个生长季，该基因表达显示明显的双锋特征，该表达模式与木材早材和晚材的发育时期相吻合，表明分离的毛白杨4CL基因与木质素的生物合成密切相关。农杆菌介导法将反义4CL基因导入烟草和毛白杨，利用分子生物学检测手段对转化植株进行筛选，获得批量转基因植株。Klason木质素含量测定分析表明，抑制内源4CL基因表达，能有效降低转基因植物中的木质素含量，且不影响植株正常生长和发育以及碳水化合物的合成。转基因毛白杨的茎杆上一些区域呈红棕色，颜色的深度与转基因毛白杨木质素含量的下降幅度呈一定的正相关性，颜色变化可作为转基因植株筛选的一个辅助指标。现已获得木质素含量下降10%以上的转基因株系3个，最多下降达41.73%，可供中试与制浆实验，为培育低木质素环保型毛白杨提供理论与实践依据。 3.为了优化现有的表达框架，使目的基因更有效地调节木质素的生物合成，应用PCR技术从毛白杨基因组中分离得到C4H(肉桂酸4—羟基化酶)基因启动子片段（GenBank注册号：AY351673）。GUS荧光活性分析和组织化学染色显示，该启动子在一些木质化的组织和器官中特异表达，随着组织成熟度和木质化程度的增加，表达活性逐渐增强，并且该启动子受伤诱导。反义CCoAOMT基因在C4H启动子的调控下，会引起转基因烟草木质素均有不同程度的减少，但不影响碳向碳水化合物的转换合成，对植物的生长发育也无明显负效应。这些结果证明了从毛白杨中分离的C4H 启动子可以应用于造纸原料树种材性改良的遗传工程操作。 4．首次从水稻中华10号（Oryza sativa L. ssp. japonica）分离了CCoAOMT基因家族的三个成员，对其基因结构及表达特性的分析表明，该基因家族的三个成员与水稻的木质化进程关系密切，研究结果有助于了解单子叶植物中的甲基化途径发生机制，为高产水稻抗倒伏和茎杆饲料作物的遗传改良奠定了基础。

内蒙古典型草原地上净初级生产力估测及模型模拟的研究

地上净初级生产力（ANPP）是陆地生态系统碳循环的重要组成部分，但由于估测ANPP的方法不同使得对ANPP的估测值存在很大的不确定性。本文采用3种方法（群落中所有种群当年最大地上生物量之和（ANPPC1）、当年群落最大地上生物量（ANPPC2）以及每年固定日期（8月30日）的地上生物量（ANPPC3））在种群、功能群和群落水平上分别对连续19年（1980～1998）的内蒙古羊草和大针茅草原生态系统功能（如ANPP、植物多样性和水分利用效率（WUE））的动态变化进行了比较分析，同时探讨了不同估测方法下气候和放牧对ANPP的影响，在此基础上利用DNDC模型进行了ANPP的模拟和敏感性分析研究，主要结果包括： 在羊草和大针茅草原群落中，不同主要植物种群或功能群多年平均地上最大生物量出现的时间不同，而同一植物种群或功能群的年地上最大生物量出现的时间存在年际间的变化。采用当年最大地上生物量和8月30日固定日期的地上生物量作为群落ANPP的这两种方法高估了建群种或禾草功能群在群落中的作用。 羊草草原群落多年平均ANPPC1、ANPPC2和 ANPPC3分别是257.5、190.1和166.0 g.m-2;相应的大针茅草原多年平均ANPPC1、ANPPC2和 ANPPC3分别是180.4、132.8和122.5 g.m-2。就群落生产力而言，后两种常用的方法二和方法三分别低估了草原群落ANPP 14.2%～40.0%和15.5～59.0%。本文研究表明尽管ANPPC1与ANPPC2和ANPPC3之间存在显著的差异，但二者之间存在极显著的相关性:羊草草原的ANPPC1= 59.587+1.061×ANPPC2（r2＝0.865, p＜0.001），ANPPC1= 92.329+1.017×ANPPC3（r2＝0.569, p＜0.001）;大针茅草原的ANPPC1= 32.918+1.114×ANPPC2（r2＝0.814, p＜0.001），ANPPC1= 76.120+0.875×ANPPC3（r2＝0.499, p＝0.001）。 种群、功能群和群落地上净初级生产力与气候因子间的关系因不同的估测方法而异。羊草草原的建群种羊草种群仅ANPPS3与8月和11月份的平均温度间存在显著相关性，而大针茅草原群落的建群种大针茅种群的ANPPS1和ANPPS2与3月份平均最高气温间呈负相关关系。在羊草草原群落，杂类草功能群ANPPF1和ANPPF2与3月份气温呈负相关关系;而灌木半灌木功能群ANPPF1和ANPPF2与3月份降水呈负相关关系。在大针茅草原群落，禾草功能群ANPPF1与5月份最高平均气温呈显著负相关关系。羊草草原群落ANPPC1和ANPPC3分别与11月份最低温度和平均温度存在显著负相关关系;大针茅草原群落ANPPC1与2和6月份月降水量间相关性显著，ANPP2与5月和11月份月平均最低温度呈显著负相关关系，而4～9月份，1和4月份平均最低温度对群落ANPPC3起决定作用。 在羊草和大针茅草原群落，由方法一得到的群落水分利用效率、Shannon植物多样性指数和均匀度指数与方法二或方法三得到的相应的指数间存在显著的差异，方法二和方法三得到的值间差异不显著。群落地上净初级生产力与相应估测方法的植物Shannon多样性指数和均匀度指数之间相关性不显著。 放牧条件下羊草或大针茅草原群落中的建群种羊草和大针茅在群落中的相对地上生物量较围栏内的相应植物种群的降低，而糙隐子草种群在群落中的比例上升。不同的放牧管理条件下，群落中植物种群地上现存量的季节动态发生变化。群落的植物组成及植物种群地上生物量在群落中总生物量的比例发生了明显的变化。利用遥感来估测地上净初级生产力时，分别低估了羊草和大针茅草原ANPP 52％和27％。 DNDC模型可以很好地模拟内蒙古典型草原生态系统的地上生物量，通过敏感性分析表明降水是草原植物生长的主要限制因子，在降水量增加或降低至日降水的30%时，模拟的地上生物量显著地高于或低于实测地上生物量值。在多年平均降水量为347mm的情况下，随着土壤粘粒含量的增加，地上生物量逐渐降低，与北美草原一致。

PHYLOGENY OF RHESUS-MONKEYS (MACACA-MULATTA) AS REVEALED BY MITOCHONDRIAL-DNA RESTRICTION ENZYME ANALYSIS

Mitochondrial DNA, purified from 36 samples of 23 local populations which are widely distributed in Vietnam, Burma, and 10 provinces of China, has been analyzed to model the phylogeny of rhesus monkeys. The 20 local populations of China may represent nearly all major populations in China. Using 20 restriction endonucleases of 6-bp recognition, we observed a total of 50-61 sites in the various samples. By combining the cleavage patterns for each enzyme, the 36 samples were classified into 23 restriction types, each of which was found exclusively in the respective population from which samples were obtained By combining the earlier study of Indian rhesus monkeys, phylogenetic trees, which have been constructed on the basis of genetic distance, indicate that rhesus monkeys in China, Vietnam, India, and Burma can be divided into seven groups. Integrating morphological and geographical data, we suggest that rhesus monkeys in China, Vietnam, and Burma may be classified into six subspecies-M. m. mulatta, M. m. brevicaudus, M. m. lasiotis, M. m. littoralis, M. m. vestita, and M. m. tcheliensis-and rhesus monkeys in India may be another valid subspecies. M. m. tcheliensis is the most endangered subspecies in China. Divergence among subspecies may have begun 0.9-1.6 Ma. The radiation of rhesus monkeys in China may have spread from the southwest toward the east. The taxonomic status of the Hainan monkey and the Taiwan monkey require further investigation.

溶藻细菌DC21的分离、鉴定及其溶藻特性

从云南滇池中分离得到一株溶藻细菌DC21,经鉴定为葡萄球菌属的1种(Staphylococcussp.),可溶解铜绿微囊等9种藻.试验表明,经过滤、离心沉降及高温灭菌处理的滤液,均能强烈抑制铜绿微囊藻的生长,说明其溶藻作用的因子之一为该菌的胞外分泌物,且这种物质具有热稳定性.细菌DC21使藻的光合作用受到抑制;在试验的第1d,受试藻的超氧物歧化酶及过氧化氢酶活性显著增高,但在第7d,这两种抗氧化保护酶的活性显著下降;受试藻的丙二醛含量明显增高,说明该菌促使其发生了膜脂过氧化.

淡水鱼类粘孢子虫的18S rDNA分子系统学研究

利用两个通用引物myxoF(5′ CGCGGTAATTCCAGCTCCAGTAG 3′)和myxoR(5′ ACCAGGTAAGTTTTCCCGTGTTGA 3′)成功扩增出圆形碘泡虫、全圆碘泡虫、武汉单极虫、微山尾孢虫和库班碘泡虫 5种粘孢子虫的部分 18SrDNA序列 ,其GenBank登录号为 :AY16 5 179—AY16 5 183。并结合GenBank其他 13个相关序列构建了 18个物种的分子系统树。结果表明 ,碘泡虫 ,尾孢虫和单极虫较Tetracapsulabryozoides和“PK

三个鲤品种线粒体基因片段序列保守性

在红鲤、镜鲤和野鲤中进行线粒体 1 6SrRNA和Cytb基因片段的序列测定。结果发现 ,3个鲤品种在多达 859个碱基长度上无变异 ,但是与已报道的鲤全序列中的同源序列则有约 1 5%的趋异。这个结果提示 3个鲤品种可能在起源中是独立的一支 ,并且分化极低

The role of NH4+ toxicity in the decline of the submersed macrophyte Vallisneria natans in lakes of the Yangtze River basin, China

Experimental and field studies were conducted to evaluate the effects of NH4+ enrichment on growth and distribution of the submersed macrophyte, Vallisneria natans L, in lakes of the Yangtze River in China, based on the balance between free amino acids (FAA) and soluble carbohydrates (SC) in the plant tissue. Increase of NH4+ rather than NO3- concentrations in the water column caused FAA accumulation and SC depletion of the plant. The plant showed a unimodal pattern of biomass distribution along both FAA/SC ratios and external NH4+ concentrations, indicating that a moderate NH4-N concentration (< 0.3 mg L-1) benefited the plant, whereas the high NH4-N concentration (> 0.56 mg L-1) eliminated the plant completely. Therefore, 0.56 mg NH4-N mg L-1 in the water column was taken as the upper limit for V. natans in lakes of the Yangtze River basin. The mesocosm experiment showed that at a high external NH4-N (0.81 mg L-1), V. natans failed to propagate with a loss of half SC content (5 mg g(-1) DW) in the rhizomes, indicating that the consumption of carbohydrates for detoxification of excess NH4+ into non-toxic FAA significantly diminished carbohydrate supply to the rhizomes. This might consequently inhibit the vegetative reproduction of the plant, and also might be an important cause for the decline and disappearance of the plant with eutrophication. The present study for the first time reports substantial ecophysiological evidences for NH4+ stress to submersed macrophytes, and indicates that NH4+ toxicity arising from eutrophication probably plays a key role in the deterioration of submersed macrophytes like V. natans.

Effects of desiccation and CO2 concentrations on emersed photosynthesis in Porphyra haitanensis (Bangiales, Rhodophyta), a species farmed in China

The effects on photosynthesis of CO, and desiccation in Porphyra haitanensis were investigated to establish the effects of increased atmospheric CO2 on this alga during emersion at low tides. With enhanced desiccation, net photosynthesis, dark respiration, photosynthetic efficiency, apparent carboxylating efficiency and light saturation point decreased, while the light compensation point and CO2 compensation point increased. Emersed net photosynthesis was not saturated by the present atmospheric CO2 level (about 350 ml m(-3)). and doubling the CO2 concentration (700 ml m(-3)) increased photosynthesis by between 31% and 89% at moderate levels of desiccation. The relative enhancement of emersed net photosynthesis at 700 ml m(-3) CO2 was greater at higher temperatures and higher levels of desiccation. The photosynthetic production of Porphyra haitanensis may benefit from increasing atmospheric CO2 concentration during emersion.

Phylogenetic relationships of the subclass Peritrichia (Oligohymenophorea, Ciliophora) with emphasis on the genus Epistylis, inferred from small subunit rRNA gene sequences

The peritrichs have been recognized as a higher taxon of ciliates since 1968. However, the phylogenetic relationships among them are still unsettled, and their placement within the class Oligohymenophorea has only been supported by the analysis of the small subunit rRNA gene sequence of Opisthonecta henneguyi. DNA was isolated directly from field-sampled species for PCR, and was used to resolve relationships within the genus Epistylis and to confirm the stability of the placement of peritrichs. Small subunit rRNA gene sequences of Epistylis plicatilis, Epistylis urceolata, Epistylis chysemydis, Epistylis hentscheli, Epistylis wenrichi, and Vorticella campanula were sequenced and analyzed using both distance-matrix and maximum-parsimony methods. In phylogenetic trees, the monophyly of both the genus Epistylis and the subclass Peritrichia was strongly supported, while V. campanula clustered with Vorticella microstoma. The topology in which E. plicatilis and E. hentscheli formed a strongly supported sister clade to E. urceolata, E. chrysemydis, and E. wenrichi was consistent with variations in the thickness of the peristomial lip. We concluded that the peristomial area, especially the. peristomial lip, might be the important phylogenetic character within the genus Epistylis.

High performance 1689-nm quantum well diode lasers

1689-nm diode lasers used in medical apparatus have been fabricated and characterized. The lasers had pnpn InP current confinement structure, and the active region consisted of 5 pairs of InGaAs quantum wells and InGaAsP barriers. Stripe width and cavity length of the laser were 1.8 and 300 pm, respectively. After being cavity coated. and transistor outline (TO) packaged, the lasers showed high performance in practice. The threshold current was about 13 +/- 4 mA, the operation current and the lasing spectrum were about 58 6 mA and 1689 +/- 6 nm at 6-mW output power, respectively. Moreover, the maximum output power of the lasers was above 20 mW.

Research on nitrogen implantation energy dependence of the properties of SIMON materials

With different implantation energies, nitrogen ions were implanted into SIMOX wafers in our work. And then the wafers were subsequently annealed to form separated by implantation of oxygen and nitrogen (SIMON) wafers. Secondary ion mass spectroscopy (SIMS) was used to observe the distribution of nitrogen and oxygen in the wafers. The result of electron paramagnetic resonance (EPR) was suggested by the dandling bonds densities in the wafers changed with N ions implantation energies. SIMON-based SIS capacitors were made. The results of the C-V test confirmed that the energy of nitrogen implantation affects the properties of the wafers, and the optimum implantation energy was determined. (c) 2005 Elsevier B.V. All rights reserved.

Selective-area MOCVD growth for distributed feedback lasers integrated with vertically tapered self-aligned waveguide

The growth pressure and mask width dependent thickness enhancement factors of selective-area MOCVD. grow th were investigated in this article. A, high enhancement of 5.8 was obtained at 130 mbar with the mask width of 70 mum. Mismatched InGaAsP (-0.5%) at the maskless region which could ensure the material at butt-joint region to be matched to InP was successively grown by controlling the composition and mismatch modulation in the selective-area growth. The upper optical confinement layer and the butt-coupled tapered thickness waveguide were regrown simultaneously in separated confined heterostructure 1.55 gm distributed feedback laser, which not only offered the separated optimization of the active region and the integrated spotsize converter, but also reduced the difficulty of the butt-joint selective regrowth. A narrow beam of 9degrees and 12degrees in the vertical and horizontal directions, a low threshold current of 6.5 mA was fabricated by using this technique. (C) 2003 Elsevier Science B.V. All rights reserved.