36 resultados para Cholestérol 7a-hydroxylase


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An improved 2 ×2 silicon-on-insulator Mach-Zehnder thermo-optical switch is designed and fabricated, which is based on strongly guided multimode interference couplers and single- mode phase-shifting arms. The multimode interference couplers and input/output waveguides are deeply etched to improve coupler performances and coupler-waveguide coupling efficiencies. However, shallow etching is used in the phase-shifting arms to guarantee single-mode property. The strongly guided coupler presents an attractive uniformity about 0. 03 dB and a low propagation loss of -0.6 dB. The 2× 2 switch shows an insertion loss as low as -6.8 dB, where the fiber-waveguide coupling loss of -4.3 dB is included, and the response-time is measured as short as 6.8 μs, which are much better than our previous results.

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设计并制作了一种新型的SOI 2×2马赫-曾德(MZ)热光开关.这种光开关采用了深刻蚀结构的配对多模干涉耦合器,同时,为了保证单模传输和调制,在连接波导和调制臂区域采用了浅刻蚀结构.深刻蚀结构增强了多模干涉耦合器对光场的限制,有利于自映像质量的提高,从而减少了自映像损耗和不均衡度,同时也提高了制作容差.基于强限制配对干涉耦合器的新型热光开关,其插入损耗为-11.0 dB,其中包括光纤-波导耦合损耗-4.3 dB,上升和下降开关时间分别为3.5μs和8.8μs.

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本论文由三章组成。第一章阐述了藏药水菖蒲的化学成分研究,共分离鉴定了39个化学成分,其中6个为新化合物。第二章报道了几种忍冬属植物的HPLC、HPLC-MS、GC分析以及抑菌活性、重金属含量测定结果。第三章概述了菖蒲属植物的研究进展。 第一章报道了水菖蒲(Acorus calamus L.)化学成分的分离纯化与结构鉴定。采用正、反相硅胶柱层析等分离方法,从水菖蒲的根中共分离出41个化合物,通过红外、质谱、核磁共振及X-ray单晶衍射等波谱方法和模拟计算方法鉴定了其中39个化合物的结构,主要为倍半萜、苯丙素、甾体类化合物。其中含有5个新的倍半萜类化合物和1系列新的甾体皂苷衍生物。经波谱分析将它们的结构鉴定为 1b, 7a(H)-cadinane-4a, 6a, 10a-triol (1), (2R,6R,7S,9S)-1(10), 4-cadinadiene-2, 9-diol (2), 1a, 5b-guaiane-10a-O-ethyl-4b, 6b-diol (7), 6b, 7b(H)-cadinane-1a, 4a, 10a-triol (13),(1R,4R,6S,10R)-1-hydroxy-7(11)-cadinen-5, 8-dione (14), 4′-O-正n碳酰基-3-O- β-D-葡萄糖基谷甾醇(n=14, 16, 18, 22) (15)。 第二章包括四个部分。第一部分报道了忍冬属三种植物40个样品的HPLC测定和对主要活性成分绿原酸的定量分析结果,以及运用HPLC-MS技术对色谱图中8个峰进行指认。在此基础上,考察了种植和采收多个因素对绿原酸含量的影响。第二部分报道了忍冬属三种植物27个样品的GC分析,根据样品的挥发性成分的保留时间对不同样品进行了定性比较,并考察了花期及海拔高度对植物挥发性成分的影响。第三、四部分分别阐述了灰毡毛忍冬和红腺忍冬的体外抑菌活性研究和重金属含量测定结果。 第三章全面系统地概述了菖蒲属植物的化学成分和药理活性研究进展。 This dissertation is composed by three chapters. The first chapter elaborates the phytochemical investigation of Acorus calamus L. Thirty-nine compounds including six new compounds were isolated and identified. The second chapter reports the research on genus Lonicera by HPLC, HPLC-MS and GC. Antifungal activity and heavy metals measurement of genus Lonicera were reported. The third chapter is a review about the research progress on the plant family of Acorus. The first chapter focuses on the isolation and identification of chemical constituents from Acorus calamus L.. Forty-one compounds were isolated from the root of Acorus calamus L. by repeat column chromatography over normal and reversed phase silica gel, the structure of thirty-nine compounds was identified by spectroscopic methods and computational methods, including IR, MS, NMR and X-ray. Those compounds mainly belonged to sesquiterpene, phenylpropanoid and steroid. Among them, five are new sesquiterpenes and one series are new steroid glycoside derivatives. Their structure were suggested as 1b, 7a(H)-cadinane-4a, 6a, 10a-triol (1), (2R,6R,7S,9S)-1(10), 4-cadinadiene-2, 9-diol (2), 1a, 5b-guaiane-10a-O-ethyl-4b, 6b- diol (7), 6b, 7b(H)-cadinane-1a, 4a, 10a-triol (13), (1R,4R,6S,10R)-1-hydroxy-7(11)- cadinen-5, 8-dione (14), 4′-O-carbonyl-3-O-β-D-glucosyl-sitosterol (carbonyl = tetradecanoyl, hexadecanoyl, octadecyl, docosanoyl) (15). The second chapter consists of four parts. The first part reports the HPLC analysis of forty samples of the genus Lonicera, and the quantitative investigation of chlorogenic acid in these samples by HPLC analysis. Relationship between the content of chlorogenic acid in different samples and their planting conditions and harvesting time were discussed. Furthermore, eight compounds were identified or tentatively characterized based on their mass spectra and UV spectra profiles. The second part is about qualitative analysis of the volatile constituent in twenty-seven samples of genus Lonicera by GC. The effect of planting altitude and harvesting time on the volatile constituent was also investigated. The third and fourth parts describe the antifungal activity and content of some kinds of heavy metals of L. macranthoides Hand.-Mazz. and L. hypoglauca Miq.. The third chaspter is a review about the research progress of the plant family of Acorus.

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本学位论文由两部分共3章组成。第一部分分别报道2种藏药唐古特瑞香和大果大戟化学成分的研究成果,从2种药用植物共分离鉴定了60个不同的化合物,其中12个为新结构,特别有意义的是发现了2个具有同一新骨架的二萜化合物。第二部分概述了大戟科植物多环二萜的研究进展。 第一部分包括第1和2章。在这2章中分别报道了唐古特瑞香(Daphne tangutica Maxim)和大果大戟(Euphorbia wallichii Hook. f. Fl)化学成分的分离纯化与结构鉴定。实验采用正、反相硅胶柱层析、薄层制备及HPLC等分离方法,从唐古特瑞香的根皮中共分离出32个化合物,通过红外、质谱及核磁共振等波谱方法鉴定了其中的31个,结构类型分别属于瑞香二萜类、木脂素类、香豆素类、苯丙素类及甾体类,其中有三个新的瑞香二萜型化合物,经波谱分析将它们的结构分别鉴定为1,2a-二氢-20-棕榈酰瑞香毒素、1,2a-二氢-5b-羟基-6a,7a-环氧-树脂大戟醇-14-苯甲酸酯及1,2b-二氢-5b-羟基-6a,7a-环氧-树脂大戟醇-14-苯甲酸酯,另外还有13个已知化合物为首次从该植物中分离得到。从大果大戟的根部共分离出33个化合物,鉴定了其中的30个,其主要成分为种类丰富的二萜,包括巨大戟烷型、续随子烷型、对映-阿替生烷型、对映-贝壳杉烷型、对映-松香烷型、ent-trachylobane型、对映-异海松烷型及一新骨架五环二萜ent-wallichane型,另外还有香豆素、甾体、三萜和一些简单的小分子化合物。其中新化合物有9个,经波谱分析将它们的结构分别鉴定为5-O-(2E,4E,6E)-癸三烯酰基-3,20-O-二乙酰基巨大戟醇、5-O-(2E,4Z)-癸二烯酰基-3-O-乙酰基-20-去氧巨大戟醇、3-O-(2E,4Z)-癸二烯酰基-5b,6b-氧-交京大戟醇、7-苯甲酰氧基-3,5,15-三乙酰基-续随子醇、ent-trachylobane-3-one-17-oic-acid、3α-羟基-对映-阿替斯-16-烯-14-酮、3α,6-二羟基-对映-异海松-7-烯-2,15-二酮、wallichanol A 和 wallichanol B,其中,wallichanol A 和 wallichanol B属于一新骨架类型的五环二萜。除此以外,另有13个已知化合物为首次从该植物中分离得到。 第2部分即第3章,首次概述了大戟属植物中多环二萜的化学和药理研究进展。 This dissertation is composed by two parts. The first part reports the phytochemical investigation of two Tibetan medicine plants, Daphne tangutica Maxim and Euphorbia wallichii Hook. f. Fl. Sixty different compounds including ten new compounds and two novel diterpenoids possessing a new carbon skeleton were isolated and identified. The second part is a review about the progress of studies on the polycyclic diterpeniods of the plant family of Euphorbia. The first part consists two chapters, which expatiate on the isolation and identification of chemical constituents from D. tangutica and E. wallichii. Thirty-one compounds were isolated from the root barks of D. tangutica by methods of column chromatography (silica gel, including reversed phase), preparative TLC and HPLC, and their structure were identified as nine daphnane diterpenes, six lignans, nine cumarin derivatives, five phenylpropanoid derivatives, a steroids and a benzoate on the basis of spectroscopic methods including IR, MS and NMR. Among them, three are new diterpenes with skeleton of daphnane and the structure were determined as 1,a-dihydro-20-palimoyl-daphnetoxin, 1,2a-dihydro-5b- hydroxy-6a,7a-epoxy-resiniferonol-14-benzoate and 1,2b-dihydro-5b-hydroxy- 6a,7a-epoxy-resiniferonol-14-benzoate. In addition, thirteen known ones were isolated from this plant for the first time. Isolation of the roots of E. wallichii yielded thirty compounds, twenty-four of them were elucidated as diterpenoids, which belong to different skeleton types of ingenol, lathyrane, ent-atisane, ent-kaurane, ent-abietane, ent-trachylobane, ent-isopimarane and a new pentacyclic skeleton ent-wallichane respectively. The remains including a cumarine, a triterpenoid, a steroid and three compounds with small molecule. Nine new compounds were characterized as 5-O-deca-2E,4E,6E- trienoyl-3,20-O-diacetylingenol, 5-O-deca-2E,4Z-dienoyl-3-O-acetyl-20- deoxyingenol, 3-O-deca-2E,4Z-dienoyl-jolkinol-5b,6b-oxide, 7-benzoyl-3,5,15- triacetyl-7-hydroxylathyrol, ent-trachylobane-3-one-17-oic-acid, 3α-hydroxy-ent- atis-16-en-14-one, 3α,6-dihydroxy-ent-isopimarane-7-en-2,15-dione, wallichanol A and wallichanol B, respectively, by means of comprehensive spectroscopic analysis. Among them, wallichanol A and wallichanol B were two notable novel pentacyclic diterpenoids processing a new rearranged carbon skeleton. And more, thirteen ones were firstly reported from this plant. The third chapter summarizes the research development on chemistry and pharmacology of polycyclic diterpenes from the plant family of Euphorbia for the first time.

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本论文由三章组成。第一、二章分别报道钩藤、蹄叶橐吾的化学成分研究工作。从两种药用植物中共分离和鉴定了37 个化合物,其中有1 组6 个新的骨架相似的鞘糖脂类同系物及5 个新的倍半萜类化合物。第三章概述了艾里莫芬烷内酯类化合物的研究进展。 第一章报道钩藤(Uncaria rhynchophylla)带钩茎枝乙醇提取物的化学成分。采用正、反相硅胶柱层析等分离方法,运用NMR、MS 等波谱技术共分离鉴定得到22 个化合物,分属于五环三萜类化合物、鞘糖脂类化合物等,其中有1 组6个新的鞘糖脂类化合物同系物,另外有7 个化合物首次从该植物中分离得到。 第二章报道蹄叶橐吾(Ligularia fischeri)根部乙醇提取物化学成分的分离纯化和结构鉴定。通过正、反相硅胶柱层析等分离纯化和MS、NMR 等波谱解析,共分离鉴定得到18 个化合物,其中5 个是新化合物。它们的结构分别确定为3β-acetyl-6β, 8α, 10β-trihydroxyeremophila-7(11)-en-8, 12-olide (23), 3β-acetyl-8β,10β-dihydroxy-6β-(2-methylbutyryloxy)-eremophilenolide (24), 3β-acetyl-6β, 10β-dihydroxyeremophila-7(11), 8(9)-dien-8, 12-olide (25), 3β-acetyl-6β, 10β-dihydroxyeremophila- 7(11), 8(9)-dien-8, 12-olide (26), (3aR, 4R, 5S, 7aS)-2-acetyl-7ahydroxy-3a, 4, 5, 6, 7, 7a-hexahydro-1H-inden-5-yl acetate (27)。这5 个化合物经体外生物活性测试,结果表明化合物23 具有抑制酪氨酸磷酸酯酶的活性, 其IC50 值为1.30 μg/ml。 第三章概述了近二十年来艾里莫芬烷内酯类化合物的研究进展,包括其结构及药理活性两个方面,列举了从23 种植物中分到的114 个艾里莫芬烷内酯类化合物,同时也对其主要生物活性进行了总结。 This dissertation consists of three chapters. The first and second chapters elaborate the phytochemical investigation of Uncaria rhynchophylla and Ligularia fischeri. Thirty-seven compounds, including six new ones, were isolated and identified by spectroscopic methods and X-ray diffraction experiments. The third chapter is a review on the study progress of Eremophilanolides. The first chapter focus on the isolation and identification of chemical constituents from Uncaria rhynchophylla. Twenty-two compounds were isolated from the roots of U. rhynchophylla by repeat column chromatography over normal and reversed phase silica gel. Those compounds mainly belonged pentacyclic triterpenoids and sphingolipids. Among them, a series homologues of six sphingolipids were new compounds and seven compounds were firstly reported in this plant. The second chapter is about the phytochemical investigation of L. fischeri. Eighteen compounds were isolated and identified. Among them, four new erem-ophilanolides and a new dinoreremophilane derivative were characterized as 3β-acetyl-6β, 8α, 10β-trihydroxyeremophila-7(11)-en-8, 12-olide (23), 3β-acetyl-8β, 10β-dihydroxy-6β-(2-methylbutyryloxy)-eremophilenolide (24), 3β-acetyl-6β, 10β-dihydroxyeremophila-7(11), 8(9)-dien-8,12-olide (25), 3β-acetyl-6β, 10α-dihydroxyeremophila- 7(11), 8(9)-dien-8,12-olide (26), (3aR,4R,5S,7aS)-2-acetyl-7a-hydroxy-3a, 4, 5, 6,7, 7a-hexahydro-1H-inden-5-yl acetate (27) by spectroscopic analysis and confirmed by X-ray crystallography analysis. It was revealed that compound 23 has the ability to inhabit the PTP1B in vivo. And its IC50 is 1.30 μg/ml. The third part is a review on the study progress of Eremophilanolides.

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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.

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赤霉素是一种高效能的广谱植物生长调节剂,为五大植物激素之一,具有重要的生物学功能。目前利用赤霉素突变体研究生物合成途径和信号转导已经成为热点。 GA 20-氧化酶是GA生物合成中的一类关键酶,它位于GA合成途径的中心位置。本研究根据烟草(Nicotiana tabacum)GA 20-氧化酶基因序列,设计2对分别含有特定酶切位点的特异引物,以烟草基因组DNA为模板,扩增目的基因(约250 bp)片段。将正、反向目的片段分别插入中间载体的内含子两侧,再经BamH I和Sac I双酶切回收约700 bp的目的片段,插入到双元载体质粒p2355中,成功构建了含GA 20-氧化酶基因片段反向重复序列的植物表达载体p23700。分别将p2355质粒和p23700质粒导入根癌农杆菌(Agrobacterium tumefaciens)EHA105中并转化烟草叶片细胞,经卡那霉素选择培养,PCR及GUS组织染色鉴定,获得转基因烟草植株。以EHA105-p2355转化的烟草,获得41株转基因植株,均没有矮化表型;而以EHA105-p23700转化的烟草,获得转基因植株14株,其中具有矮化表型的烟草10株,表明反向重复序列转录产物能形成发夹RNA(hpRNA),产生小分子干扰RNA(small interferring RNA,简称siRNA),干扰目的基因的表达。 赤霉素含量测定表明矮化植株中赤霉素合成途径的最终产物GA3总含量明显低于野生型烟草植株。荧光定量PCR结果表明,矮化转基因烟草的GA 20-氧化酶基因表达量受到明显抑制,表达量明显低于野生型对照。同时对上游内根-贝壳杉合成酶(Ent-kaurene synthase,KS)基因,下游的GA-3β羟化酶基因进行了RT-PCR分析,结果显示上游基因的表达没有规律性变化,而下游基因表达量亦降低。上述结果表明,GA 20-氧化酶基因的表达被有效地干扰了,表达受到抑制,从而影响植株体内GA3的合成,影响植株的生长发育,导致植株矮化。并推测,GA 20-氧化酶基因受到抑制,可能影响下游基因的表达。并且通过干旱胁迫测试,发现矮化植株相对于野生型植株及不含干扰片段的转基因植株,对干旱的耐受力有了很大的提高,具有更强的耐受力。 研究结果为进一步进行相关研究奠定基础。 Gibberellin(GA) is an efficient plant growth regulator. As one of five major plant hormones, it plays an important biological function. Using GA mutant for investigating biosynthetic pathways and signal transduction has become high lights. GA 20-oxidase is a crucial enzyme involved in gibberellin biosynthesis. According to tobacco (Nicotiana tabacum) GA 20-oxidase enzyme gene sequence and based on binary vector p2355, we constructed a plant expression vector p23700, which habors an inverted repeat DNA fragment of GA 20-oxidase gene drivered by Cauliflower mosaic virus promtor (CaMV 35Sp). Binary plasmid p2355 had no inverted repeat DNA fragment of GA 20-oxidase gene. The vector p2355 and p23700 were introduced into Agrobacterium tumefaciens EHA105 and tobacco leaf transformation was conducted. After selected by kanamycin and characterized by PCR and GUS hischemical reaction, transsgenic plants were obtained. Fourtheen transgenic plants, which were transformed by EHA105-p23700, were obtained. Among them, 10 were dwarf mutants. However, 41 transgenic plants with the same normal phenotype as wild type,which were transformed by EHA105-p2355, were obtained. Analysis of Gibberellin contents showed that it was lower in dwarf mutants than in normal phenotype plants. Moreover, comparing to normal phenotype plants including wild type and transgenic plants with no interference fragment, the drought tolerance of dwarf plants have greatly increased. And their proline content increased obviously after drought test. Fluorescence quantitative real time PCR (RT-PCR) showed that GA 20-oxidase gene expression was significantly inhibited in dwarf transgenic tobacco. Meanwhile, the expression of the upstream gene ent-kaurene synthase (KS) gene and downstream gene GA-3β hydroxylase gene was also detected by RT-PCR. The results presented that KS gene expression had no regular change while GA-3β hydroxylase gene expression reduced. It implied that inhibiting GA 20-oxidase gene probably reduce the expression of downstream genes. The results showed that the transcriptional products of the foreign inverted repeat fragment can form hairpin RNA (hpRNA) to induce RNAi. It presented that GA 20-oxidase gene expression was effectively interfered, resulting in reducing GA3 synthesis and inhibiting plant growth and development, then dwarf plants were produced. However, the dwarf plants had higher tolerance of drought.

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研究了长白山北坡椴树 ( Tilia amurensis)枝条的分解进程。实验结果揭示 ,分解率是与各实验地的植被类型 ,海拔 ,生长季月平均大于 5℃的积温等密切相关。其枝条分解的保存率 ( Y)与分解年龄 ( x)的相关 ,可用指数模型 :Y=e- kx来拟合。在红松阔叶混交林 ,红松针叶混交林 ,岳桦云冷杉林及岳桦林中椴树枝条分解常数 K分别为 -0 .1 6 8与 -0 .1 2 7,-0 .1 0 2和 -0 .0 94。枝条原重量 5 0 %的分解年龄在上述 4个森林中分别是 4 ,5 ,6和 7a。实验还指出 ,在长白山红松阔叶混交林皆伐迹地的实验条件下适当的蔽荫和浇水对分解有正面影响 ,而且 ,枝条的分解率主要是以长白山不同海拔植被类型的温度条件而转移 ,特别是生长季的积温

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实地测定了黄土高原半干旱区固原不同生长年限苜蓿草地和连作8a苜蓿草地翻耕轮作不同年限粮食作物后深层土壤水分特征,分析了苜蓿草地土壤干燥化特征和粮草轮作对土壤水分的恢复效应。结果表明:(1)苜蓿连作1a、5a、8a和12a等4类苜蓿草地0~1000cm土层平均土壤湿度值为6.6%,平均土壤水分过耗量702.8mm,平均土壤干燥化速率147.1 mm/a,达到强烈干燥化程度,苜蓿连作5a土壤干层深度超过1000cm,苜蓿连作8a土壤干层深度超过1360cm,苜蓿草地合理利用年限为7a。(2)连作8a苜蓿草地翻耕并轮作4~7a和25a粮食作物等5类粮田0~1000cm土层土壤湿度介于6.74%~11.95%,土壤贮水量恢复值介于210.6~887.3mm,平均土壤水分恢复速率为80.8mm/a。轮作6a后粮田土壤干层轻度恢复程度以上深度达到1000cm。通过粮草轮作使苜蓿草地土壤湿度恢复到当地土壤稳定湿度需要13a以上。黄土高原半干旱区适宜的粮草轮作模式为:7a苜蓿→13a粮食作物。

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以在陕北丘陵沟壑区坡耕地不同耕作法试验 7a的资料为依据 ,论述了不同耕作法的水分利用、土壤水分平衡及单位面积产量和水土保持等综合效益。作物生长主要依赖生育期内自然降雨 ;作物耗水量与每年生育期降雨量及土壤供水量呈显著正相关 ,不同耕作法之间变化不大 ,作物主要利用 0— 12 0 cm土层内的水分。同一年内不同耕作法的综合效益从优到劣、由高到低的排列顺序为宽梯田 >窄梯田 >水平沟 >平播 >水平阶 >隔坡梯田。而水土保持效益排列顺序为 :梯田 >水平沟 >隔坡梯田 >水平阶 >平播 >裸地。认为坡耕地确实应尽快退耕还林还草 ,以加速陕北生态环境建设 ,推进农业结构的战略性调整。

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Deprotonation of (ArNHPPh2NAr2)-N-1 (H[NPN](n), n = 1 - 10) by Ln(CH2SiMe3)(3)(THF)(2) (Ln = Lu, Y, Sc, Er) generated a series of rare-earth metal bis(alkyl) complexes [NPN](n)Ln(CH2SiMe3)(2)(THF)(2) (1-10), which under activation with [Ph3C][B(C6F5)(4)] and AliBu(3) were tested for isoprene polymerization. The correlation between catalytic performances and molecular structures of the complexes has been investigated. Complexes 1-5 and 8, where Ar-1 is nonsubstituted or ortho-alkyl-substituted phenyl, adopt trigonal-bipyramidal geometry. The Ar-1 and Ar-2 rings are perpendicular in 1-4 and 8 but parallel in 5. When Ar-1 is pyridyl, the resultant lutetium and yttrium complexes 9a and 9b adopt tetragonal geometry with the ligand coordinating to the metal ions in a N,N,N-tridentate mode, whereas in the scandium analogue 9c, the ligand coordinates to the Sc3+ ion in a N,N-bidentate mode. These structural characteristics endow the complexes with versatile catalytic performances, With increase of the steric bulkiness of the ortho-substituents Ar-1 and Ar-2, the 3,4-selectivity increased stepwise from 81.6% for lutetium complex 1 to 96.8% for lutetium complex 6 and to 97.8% for lutetium complex 7a. However, further increase of the steric bulk of the ligand led to a slight drop of 3,4-selectivity for the attached complex 5 (95.1%).

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A universal metal-molecule-metal sandwich architecture by the self-assembly of Ag nanoparticles (NPs) and Au NPs of various shapes interconnected with 4-aminothiophenol (4-ATP) molecules was presented. These Ag NPs/4-ATP/Au NPs sandwich structures were characterized by surface enhanced Raman scattering (SERS) using an off-surface plasmon resonance condition. Enhancement factors (EF) on the order of 10(8) for 9b(b(2)) vibration mode were observed for the 4-ATP self-assembled monolayers (SAMs) in such sandwich structures. The factors are 2 orders of magnitude larger than that on the monolayer of Au NPs of various shapes under similar condition. More importantly, remarkable increase in the intensity of b(2) vibrational modes, which is characteristic of the charge transfer (CT) behavior between metal NPs and 4-ATP molecules, was observed in these sandwich structures under 1064 nm excitation. The obtained EF on these sandwich structure for 9b(b(2)) is larger than that for 7a vibration mode by a factor of similar to 10(2), demonstrating the importance of the contribution of the CT mechanism and the CT behavior of metal contacts, which play a significant role in metal-molecule-metal nanosystems.

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A sandwich structure consisting of Ag nanoparticles (NPs), p-aminothiophenol (p-ATP) self-assembled monolayers (SAMs), and Ag NPs was fabricated on glass and characterized by surface enhanced Raman scattering (SERS). The SERS spectrum of a p-ATP SAM in such sandwich structure shows that the electromagnetic enhancement is greater than that on Ag NPs assembled on glass. The obtained enhancement factors (EF) on solely one sandwich structure were as large as 6.0 +/- 0.62x10(4) and 1.2 +/- 0.62x10(7) for the 7a and 3b(b(2)) vibration modes, respectively. The large enhancement effect of p-ATP SAMs is likely a result of plasmon coupling between the two layers of Ag NP (localized surface plasmon) resonance, creating a large localized electromagnetic field at their interface, where p-ATP resides. Moreover, the fact that large EF values (similar to 1.9 +/- 0.7x10(4) and 9.4 +/- 0.7x10(6) for the 7a- and b(2)-type vibration modes, respectively) were also obtained on a single sandwich structure of Au NPs/p-ATP SAMs/Ag NPs in the visible demonstrates that the electromagnetic coupling does not exist only between Ag NPs but also between Au and Ag NPs.

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Surface-enhanced Raman scattering (SERS) activity of silver-gold bimetallic nanostructures (a mean diameter of similar to 100 nm) with hollow interiors was checked using p-aminothiophenol (p-ATP) as a probe molecule at both visible light (514.5 nm) and near-infrared (1064 nm) excitation. Evident Raman peaks of p-ATP were clearly observed, indicating the enhancement Raman scattering activity of the hollow nanostructure to p-ATP. The enhancement factors (EF) at the hollow nanostructures were obtained to be as large as (0.8 +/- 0.3)x10(6) and (2.7 +/- 0.5)x10(8) for 7a and 19b (b(2)) vibration mode, respectively, which was 30-40 times larger than that at silver nanoparticles with solid interiors at 514.5 nm excitation. EF values were also obtained at 1064 nm excitation for 7a and b(2)-type vibration mode, which were estimated to be as large as (1.0 +/- 0.3)x10(6) and (0.9 +/- 0.2)x10(7), respectively. The additional EF values by a factor of similar to 10 for b(2)-type band were assumed to be due to the chemical effect. Large electromagnetic EF values were presumed to derive from a strong localized plasmas electromagnetic field existed at the hollow nanostructures.