Effect of plant architecture on the structure of epiphytic macro invertebrate communities in a Chinese lake

We examined the effect of different plant architecture types on epiphytic macroinvertebrates of a shallow macrophyte-dominated lake in China. Macroinvertebrates were sampled from four dominant submersed macrophytes in the lake - two dissected plants (Myriophyllum spicatum L. and Ceratophyllum demersum L.) and two undissected plants (Potamogeton maackianus A. Benn. and Vallisneria spiralis L.). Macro invertebrate richness showed significant differences among four submersed macrophyte habitats, and higher density per g of dry plant were associated with dissected plants than undissected plants. The average abundance in dissected plants was as three-six times as in undissected plants. The biodiversity of epiphytic macroinvertebrates was higher in dissected plants than undissected plants. Our results suggest that dissected plants provide different habitat for macroinvertebrates than dissected plant, and this concurs with the hypothesis that the former could support more epiphytic macroinvertebrates than the latter.

Response of chlorophyll fluorescence to dynamic light in three alpine species differing in plant architecture

A study was carried out to examine the effect of dynamic photosynthetically active photon flux density (PPFD) on photoinhibition and energy use in three herbaceous species, prostrate Saussurea superba, erect-leaved S. katochaete, and half-erect-leaved Gentiana straminea, from the Qinghai-Tibet Plateau. Chlorophyll fluorescence response was measured under each of three sets of high-low PPFD combinations: 1700-0, 1400-300, and 1200-500 mu mol m(-2) s(-1), illuminating in four dynamic frequencies: 1, 5, 15, and 60 cycles per 2 h. The total light exposure time was 2h and the integrated PPFD was the same in all treatments. The highest frequency of PPFD fluctuation resulted in the lowest photochemical activity, the highest level of non-photochemical quenching, and the greatest decrease of F-v/F-m (maximal photochemical efficiency of PSII). The 5 and 15 cycles per 2h treatments resulted in higher photochemical activity than the 1 cycle per 2h treatment. The 1700-0 PPFD combination led to the lowest photochemical activity and more serious photoinhibition in all species. S. superba usually exhibited the highest photochemical activity and CO2 uptake rate, the lowest reduction of F-v/F-m,F- and the smallest fraction of energy in thermal dissipation. With similar fractions of thermal dissipation, S. katochaete had relatively less photoinhibition than G. straminea owing to effective F-o quenching. The results suggest that high frequency of fluctuating PPFD generally results in photoinhibition, which is more serious under periods of irradiation with high light intensity. (c) 2005 Elsevier B.V. All rights reserved.

Photosynthetic depression in relation to plant architecture in two alpine herbaceous species

The Qinghai-Tibet Plateau is characterized by extremely high radiation, which may induce down-regulation of photosynthesis in plants living in this alpine ecosystem. To clarify whether photoinhibition occurs in the alpine environment and to discern its underlying mechanisms, we examined photosynthetic gas exchange and fluorescence emission in response to the changes in photosynthetic photon flux density (PPFD) and leaf temperature under natural regimes for two herbaceous species: prostrate Saussurea superba and erect-leaved Saussurea katochaete from altitude 3250 m on the Qinghai-Tibet Plateau. S. superba intercepted a higher maximum PPFD and experienced much higher leaf temperature than the erect-leaved S. katochaete. S. superba exhibited a much higher light saturation point for photosynthesis than S. katochaete. Under controlled conditions, the former species had higher CO2 uptake rates and neither species showed obvious photosynthetic down-regulation at high PPFD. Under natural environmental conditions, however, apparent photoinhibition, indicated by reduced electron transport rate (ETR), was evident at high PPFD for both species. After a night frost, the photochemistry of S. katochaete was depressed markedly in the early morning and recovered by mid-day. After a frost-free night, it was high in the morning and low at noon due to high radiation. S. superba did not respond to the night frost in terms of daytime photochemical pattern. In both species, photochemical depression was aggravated by high leaf temperature and the erect species was more sensitive to high temperature. This study suggests that the high radiation on the Qinghai-Tibet Plateau is likely to induce rapidly reversible photoinhibition, which is related closely to plant architecture. Photochemistry in the prostrate species seems able to tolerate higher PPFD, without obvious suppression, than the erect species. (C) 2003 Elsevier Science B.V. All rights reserved.

OsLIC1基因参与水稻株型调控的功能研究

CCCH型锌指蛋白是进化上比较保守的一类锌指蛋白家族，其典型的氨基酸的基序为C-X7-8-C-X5-C-X3-H，其中X为任意氨基酸，这类锌指基序一般以重复的双拷贝形式存在。本论文克隆并鉴定了一个全新的、只含有一个CCCH型锌指基序的基因，利用反义RNA策略研究该基因功能，结果发现该基因的反义转基因植株表现出叶夹角增大的表型，因此我们将该基因命名为OsLIC1（Oryza sativa Lamina Increased Leaf Angle Control 1）。生物信息学分析发现该基因定位于水稻6号染色体近端粒的一端，位于BAC克隆AP004324中。OsLIC1与通常的CCCH型锌指蛋白含有多个重复的CCCH锌指基序不同，它只含有一个CCCH型锌指基序。除了CCCH锌指结构域以外，该蛋白在靠近C-端的位置还有一段丝氨酸（Ser）富集的区域，在此区域之前，还有一个在真核生物中相对保守的，以EELR为核心基序的结构域。采用基因枪将含OsLIC1-GFP融合构建的瞬时表达载体轰击入洋葱内表皮细胞，激光共聚焦显微镜观察发现OsLIC1-GFP可以定位到细胞核中。利用酵母转录激活系统发现以EELR为核心基序的结构域具有转录激活的功能。体外核酸结合活性分析显示OsLIC1蛋白可以结合双链DNA，这些结果证明OsLIC1是一个转录因子，这也是在植物中首次发现CCCH型锌指蛋白可以作为转录因子的方式调节基因的表达。 用玉米泛素启动子（Maize Ubiquitin promoter）驱动OsLIC1基因的反义表达载体转化水稻，获得的内源OsLIC1基因表达量下降的转基因植株表现出三个明显的表型：转基因植株的叶夹角增大;转基因的株高低于对照以及转基因植株的穗粒数减少。扫描电镜观察发现转基因植株叶夹角增大是由于近轴面细胞排列发生了改变以及维管束发育受阻引起的。转基因植株的Southern Blot和RT-PCR分析，结合Western Blot分析证明了转基因植株的表型与转基因事件之间的直接联系，并证明了转基因植株中内源OsLIC1在蛋白水平的确受到了抑制。采用RT-PCR技术、Promoter：：GUS和RNA in situ杂交三种方法相结合研究OsLIC1基因的表达模式，结果表明OsLIC1基因主要在叶颈、节以及分蘖原基中表达，这与转基因植株的表型相吻合，进一步证明了转基因植株的表型与基因功能之间的关系。Affymetrix 水稻全基因组芯片分析结果显示许多受油菜素内酯诱导表达的基因在转基因植株的叶颈材料中表达量上调，RT-PCR进一步验证了这一结果。由于在转基因植株中出现的叶夹角增大的表型和水稻油菜素内酯的作用相似，而基因芯片的结果又从分子水平提供了证据和线索。进一步采用RT-PCR和Promoter：：GUS相结合的方法研究OsLIC1基因对油菜素内酯的响应，结果发现OsLIC1基因可以被油菜素内酯诱导表达。而且，OsLIC1基因的反义转基因植株与野生型相比，表现出对油菜素内酯信号更敏感的响应。根据以上结果，推测OsLIC1可能是水稻油菜素内酯信号转导途径的负调控因子。水稻油菜素内酯合成和信号转导的突变体d2-1和d61-1具有直立的叶片的特征。用反义OsLIC1转基因植株以及野生型水稻与d2-1和d61-1突变体分别进行遗传杂交，结果发现在反义OsLIC1转基因植株与d2-1和d61-1突变体的杂交F1代中，都表现出叶夹角增大的表型。但是，在F2代中，在d2-1和d61-1纯合背景下，分别表现出叶夹角增大和叶片直立的表型，说明OsLIC1上位于d2-1，而d61-1则上位于OsLIC1。这一结果进一步证明了OsLIC1是通过参与水稻油菜素内酯信号调节而发挥对水稻叶夹角的调控作用。

簇毛麦（ Dasypyrum villosum）矮秆突变体的研究

株高是农作物的重要农艺性状之一，适度矮化有利于农作物的耐肥、抗倒、高产等。20世纪50年代，以日本的赤小麦为矮源的半矮秆小麦的培育和推广，使得世界粮食产量显著增长，被誉为“绿色革命”。迄今为止，已报到的麦类矮秆、半矮秆基因已达70多个，但由于某些矮源极度矮化或者矮化的同时伴随不利的农艺性状，使得真正运用于育种实践的矮源较少。因此，发掘和鉴定新的控制麦类作物株高的基因，开展株高基因定位、克隆及作用机理等方面的研究，对实现麦类作物株高的定向改良，具有重要的理论意义和应用价值。簇毛麦(Dasypyrum villosum，2n＝14，VV)是禾本科簇毛麦属一年生二倍体异花授粉植物，为栽培小麦的近缘属。本课题组在不同来源的簇毛麦杂交后代中发现了一株自然突变产生的矮秆突变体。观察分析了该突变体的生物学特性，对矮秆性状进行了遗传分析，对茎节细胞长度、花粉的活力进行了细胞学观察，考察了该突变体内源赤霉素含量及不同浓度外施赤霉素对突变体的作用，分析了赤霉素生物合成途径中的内根贝壳杉烯氧化酶（KO）和赤霉素20氧化酶(GA20ox)的转录水平，对赤霉素20氧化酶和赤霉素3-β羟化酶(GA3ox)进行了克隆和序列分析，并对GA20ox进行了原核表达和表达的组织特异性研究。主要研究结果如下：1. 该突变体与对照植株在苗期无差异，在拔节后期才表现出植株矮小，相对对照植株，节间伸长明显受到抑制，叶鞘长度基本不变。在成熟期，对照植株的平均株高为110cm，而突变株的平均株高为32cm，仅为对照植株的1/3 左右。除了株高变矮以外，在成熟后期，突变株还表现一定程度的早衰和雄性不育。I2-KI染色法观察花粉活力结果表明，对照植株花粉90%以上都是有活力的，而突变植株的花粉仅20%左右有活力。2. 突变株与对照植株的杂交F1代均表现正常株高，表明该突变性状为隐性突变。F1代植株相互授粉得到的168株F2代植株中，株高出现分离，正常株高（株高高于80cm）与矮秆植株（株高矮于40cm）的株数比为130：38，经卡方检验，其分离比符合3：1的分离比，因此推测该突变体属于单基因的隐性突变。3. 用ELISA方法检测突变株和对照植株的幼嫩种子中内源性生物活性赤霉素（GA1＋3）含量，结果表明突变株的赤霉素含量为36 ng/ml，而对照植株的赤霉素含量为900 ng/ml。对突变株外施赤霉素，发现矮秆突变株的株高和花粉育性均可得到恢复。这些结果表明该突变株为赤霉素缺陷型突变。4. 用荧光定量PCR方法比较突变株与对照植株中内根贝壳杉烯氧化酶和赤霉素20氧化酶的转录水平，结果表明突变株的KO转录水平比对照植株分别提高了6倍（苗期）和16倍（成熟期），突变株的GA20ox转录水平与对照植株在苗期无明显差异，在成熟期突变株较对照植株则提高了10倍左右。这些结果表明该矮秆突变体与赤霉素的生物合成途径密切相关，而且极有可能在赤霉素的生物合成途径早期就发生了改变。5. 以簇毛麦总基因组为模板，同源克隆了GenBank登录号为EU142950，RT-PCR分离克隆了簇毛麦的GA3ox基因cDNA全长序列，分析结果表明该cDNA全长1206bp，含完整编码区1104bp，推测该序列编码蛋白含368个氨基酸残基，分子量为40.063KD，等电点为6.27。预测的氨基酸序列含有双加氧酶的活性结构，在酶活性中心2个Fe离子结合的氨基酸残基非常保守。该序列与小麦、大麦和水稻的GA3ox基因一致性分别为98%、96%、86%。基因组序列与cDNA序列在外显子部分一致，在478-715bp和879-1019bp处分别含238bp和140bp的内含子。6. 通过RT-PCR技术克隆了簇毛麦的GA20ox基因全长，命名为DvGA20ox，GenBank登录号为EU142949。该基因全长1080个碱基，编码359个氨基酸，具有典型的植物GA20ox基因结构。该基因编码的蛋白质与小麦、大麦、黑麦草等GA20ox蛋白的同源性分别为98%，97% 和91%。该序列重组到原核表达载体pET-32a(+)上，将获得的重组子pET-32a(+)-DvGA20ox转化大肠杆菌BL21pLysS后用IPTG进行诱导表达。SDS-PAGE分析表明，DvGA20ox基因在大肠杆菌中获得了高效表达，融合蛋白分子量为55kDa。定量PCR分析表明，该基因在簇毛麦不同器官中的表达差异明显：叶片中表达水平最高，根部表达水平次之，茎部和穗中表达较弱。在外施赤霉素后，该基因的表达水平在两小时以后急剧下降，表明该基因的表达受自身的反馈调节。本研究结果认为，(1)该簇毛麦矮秆突变体为单基因的隐性突变；(2)该矮秆突变体为赤霉素敏感突变，内源赤霉素含量检测表明突变体的内源性赤霉素含量仅为对照植株的1/30；(3)荧光定量PCR结果表明突变株的赤霉素生物合成途径的关键酶基因表达水平比对照植株高，而且突变植株的赤霉素生物合成改变很可能发生在赤霉素生物合成途径的早期；(4)GA20ox有表达的组织特异性，且受到自身产物的反馈调节。 Plant height is an impotrant agronomic trait of triticeae crops.Semi-dwarf cropcultivars, including those of wheat, maize and rice, have significantly increased grainproduction that has been known as “green revolution”. The new dwarf varieties couldraise the harvest Index at the expense of straw biomass, and, at the sametime, improvelodging resistance and responsiveness to nitrogen fertilizer. Moreover, dwarf traits ofplant are crucial for elucidating mechanisms for plant growth and development aswell. In many plant species, various dwarf mutants have been isolated and theirmodles of inheritance and physiology also have been widely investigated.The causesfor their dwarf phenotypes were found to be associated with plant hormones,especially, gibberellins GAs.Dasypyrum villosum Candargy (syn.Haynaldia villosa) is a cross-pollinating,diploid (2n = 2x = 14) annual species that belongs to the tribe Triticeae. It is native toSouthern Europe and West Asia, especially the Caucasuses, and grows underconditions unfavorable to most cultivated crops. The genome of D. villosum,designated V by Sears, is considered an important donor of genes to wheat for improving powdery mildew resistance, take-all, eyespot, and plant and seed storageprotein content. A spontaneous dwarf mutant was found in D. villosum populations.The biological character and modles of inheritance of this dwarf mutant are studied.The cell length of stem cell is observed. The influence of extraneous gibberellin tothe dwarf mutant is also examined; the transcript level of key enzyme of gibberellinbiosynthesis pathway in mutant and control plants is compared. GA3ox and GA20oxare cloned and its expression pattern is researched.1. The dwarf mutant showed no difference with control plants at seedlingstage.At mature stage, the average height of control plants were 110cm and the dwarfplants were 33cm. The height of the mutant plant was only one third of the normalplants due to the shortened internodes. Cytology observation showed that theelongation of stem epidermal and the parenchyma cells were reduced. The dwarfmutant also shows partly male sterile. Pollen viability test indicates that more than80% of the pollen of the mutant is not viable.2. The inheritance modle of this dwarf mutant is studied. All The F1 plantsshowed normal phenotype indicating that the dwarfism is controlled by recessivealleles. Among the 168 F2 plants, there are 130 normal plants and 30 dwarf plants, thesegregation proportion accord with Mendel’s 3:1 segregation. We therefore proposethat this dwarf phenotype is controlled by a single recessive gene.3. Quantitative analyses of endogenous GA1+3 in the young seeds indicated thatthe content of GA1+3 was 36ng/ml in mutant plants and 900ng/ml in normal plants.The endogenous bioactive GA1+3 in mutant plants are only about 1/30 of that innormal plants. In addition, exogenously supplied GA3 could considerably restore themutant plant to normal phenotype. These results showed that this mutant wasdefective in the GA biosynthesis.4. More than ten enzymes are involved in GA biosynthesis. KO catalyzes thefirst cytochrome P450-mediated step in the gibberellin biosynthetic pathway and themutant of KO lead to a gibberellin-responsive dwarf mutant. GA20ox catalyze therate-limited steps so that their transcript level will influence the endogenous GAbiosynthesis and modifies plant architecture. The relative expression levels of genesencoding KO and GA20ox were quantified by real time PCR to assess whether thechanges in GA content correlated with the expression of GA metabolism genes andwhere the mutant occurred during the GA biosynthesis pathway. In mutant plants,the transcript levels of KO increased about 6-fold and 16-fold at the seedling stage and elongating stage respectively comparing with the normal plants. For theseedlings, there was no notable difference in the expression of GA20ox betweenmutant and normal plants. At the elongating stage, GA20ox transcript increased 10times in mutant plants, suggesting that the GA biosynthesis pathway in mutant plantshad changed from the early steps rather than the late steps.5. A full length cDNA of D. villosum gibberellin 3β-hydroxylase homology(designated as DvGA3ox) was isolated and consisted of 1206bp containing an openreading frame of 1104bp encoding 368 predicted amino acid residues. Identityanalysis showed that the gibberellin 3β-hydroxylase nucleotide sequence shared 98%,96% and 86% homology with that of wheat, barley and rice. The predicted peptidecontained the active-site Fe of known gibberellin 3β-hydroxylase and the regionhomologous to wheat, barley and Arabidopsis. The genomic clone of gibberellin3β-hydroxylase has two introns.6. The full-length cDNA of D. villosum gibberellin 20 oxidase (designated asDvGA20ox) was isolated and consisted of 1080-bp and encoded 359 amino acidresidues with a calculated mol wt of 42.46 KD. Comparative and bio-informaticsanalyses revealed that DvGA20ox had close similarity with GA20ox from otherspecies and contained a conserved LPWKET and NYYPXCQKP regions. Tissueexpression pattern analysis revealed DvGA20ox expressed in all the tissues that wereexamined and the highest expression of DvGA20ox in expanding leaves followed byroots. Heterologous expression of this cDNA clone in Escherichia coli gave a fusionprotein that about 55KD. Transcript levels of DvGA20ox dramatically reduced twohours after application of biologically active GA3, suggesting that the biosynthesis ofthis enzymes might be under feedback control.

High-Strength Composite Fibers: Realizing True Potential of Carbon Nanotubes in Polymer Matrix through Continuous Reticulate Architecture and Molecular Level Couplings

Carbon nanotubes have unprecedented mechanical properties as defect-free nanoscale building blocks, but their potential has not been fully realized in composite materials due to weakness at the interfaces. Here we demonstrate that through load-transfer-favored three-dimensional architecture and molecular level couplings with polymer chains, true potential of CNTs can be realized in composites as Initially envisioned. Composite fibers with reticulate nanotube architectures show order of magnitude improvement in strength compared to randomly dispersed short CNT reinforced composites reported before. The molecular level couplings between nanotubes and polymer chains results in drastic differences in the properties of thermoset and thermoplastic composite fibers, which indicate that conventional macroscopic composite theory falls to explain the overall hybrid behavior at nanoscale.

Visual pattern recognition network: Its training algorithm and its optoelectronic architecture

A visual pattern recognition network and its training algorithm are proposed. The network constructed of a one-layer morphology network and a two-layer modified Hamming net. This visual network can implement invariant pattern recognition with respect to image translation and size projection. After supervised learning takes place, the visual network extracts image features and classifies patterns much the same as living beings do. Moreover we set up its optoelectronic architecture for real-time pattern recognition. (C) 1996 Optical Society of America