352 resultados para Glutaraldéhyde (GA)


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预计到本世纪末,大气CO2浓度将会增加到540~970ppm,大气CO2浓度升高所引起的全球气候变化已经受到广泛的关注。植物生长依赖CO2,并且对大气CO2浓度升高在结构和生理上产生响应。目前已有大量报道,从生态系统、群落、种群、个体、器官、组织、生理以及生化等水平上研究高浓度CO2所对植物产生的影响。但是有关高浓度CO2对植物有性生殖影响的报道却很少,同时多数实验均建立在短期的生殖响应,忽视了植物在长期高CO2浓度下具有的反馈作用和CO2浓度变化对植物的驯化作用。植物有性生殖与其生态适应性和农作物籽粒产量的关系极为密切;同时,植物有性生殖特性的变化,也可作为预测植物对全球气候变化响应的重要指标之一。为此,利用高浓度CO2对植物进行长期选择实验将很有必要。研究结果将为预测未来大气CO2浓度增加的条件下陆地生态系统的演变趋势、全球变化对植物有性生殖响应的方式和机制提供新的思路和有效方法。   在本研究中,我们以模式植物拟南芥(Arabidopsis thaliana)作为实验材料,利用370和700ppm CO2对其进行连续8个世代处理,首先研究高浓度CO2对每一个世代的拟南芥有性生殖特性的影响,然后比较各个世代中各种生殖特性指标变化的规律,从细胞、组织和个体尺度上揭示拟南芥有性生殖对全球变化的响应模式。此外,在700ppm CO2处理下,我们对拟南芥叶片生理、生化以及结构的变化进行了相关研究。两部分研究结果及主要结论如下:   首先,在每一个世代中,与370ppm CO2相比较,700ppm CO2处理显著促进了拟南芥开花,缩短生长周期,增加花、角果及种子等生殖的产量,降低种子N含量,提高种子C/N比、种子千粒重以及生殖生物量所占总生物量的比例等,而对种子萌发率、角果所含种子数目以及角果长度则无显著影响。但是, 通过对相同CO2浓度处理条件下,不同世代之间的研究结果比较发现,不同世代之间相关的生殖生物学指标并无显著差异。   其次,高浓度CO2显著降低叶片气孔密度、气孔指数、气孔导度以及蒸腾速率。在高浓度CO2处理下,叶肉细胞中叶绿体数目、叶绿体宽度和表观面积、淀粉粒大小和数量、叶片和细胞壁厚度等都显著增加,但是基粒内囊体膜的数量却显著下降。叶片中碳水化合物如可溶性总糖、淀粉以及纤维素含量在高浓度CO2下分别显著增加71.9%、78.7% 和 22.3%。此外,在高浓度CO2处理下,叶片中多数激素如如吲哚乙酸(indole-3-acetic acid, IAA)、赤霉素(gibberellin, GA)、玉米素核苷(zeatin riboside, ZR)、二氢玉米素核苷(dihydrozeatin riboside, DHZR)和异戊烯基腺苷(isopentenyl adenosine, iPA)均都显著地增加,而脱落酸(abscisic acid, ABA)含量却有所下降。最后,叶片中各种矿物质元素含量如N、P、K、Ca和Mg等含量在高浓度CO2处理下也都显著下降,而C/N比增加24.8%。   以上结果表明:   (1) 在每一个世代中,700ppm CO2处理对拟南芥各种有性生殖特性具有显著的影响,但是高浓度CO2处理对植物所引起的效应在多个世代以内并不能够传递给后代,所以在多个有性生殖世代内,高浓度CO2处理对植物生长、生殖没有驯化作用。   (2) 在高浓度CO2处理下,拟南芥叶片中叶绿体超微结构的变化,可能主要是由于叶绿体中淀粉粒数量和体积大小显著增加而引起。   (3) 在高浓度CO2处理下,由于拟南芥叶片内与促进细胞分裂与伸长的激素含量显著增加,从而对拟南芥植株生长发育速率的提高起了重要的作用。   (4) 拟南芥生长在高浓度CO2条件下,其叶片中各种矿质元素含量(如N、P、K、Ca和Mg)均显著降低,究其原因可能是,第一由于叶片中碳水化合物含量的显著增加而对矿物质元素具有稀释作用;第二由于蒸腾速率下降,引起矿质元素从根部随着蒸腾流运输到地上部分的含量相应减少。   

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水稻是世界上最重要的粮食作物,也是单子叶植物的模式植物,它为全球近一半的人口提供食物,但是低温、高盐、干旱等非生物胁迫,每年都会在全世界范围内造成水稻大面积减产。G蛋白介导的信号途径是传递胞外信号比较保守的作用机制之一。动物细胞对于G蛋白及其受体(GPCRs)的研究已经取得了很大的进展。而植物细胞中对它们的研究刚刚起步。本文从越冬稻低温响应芯片上筛选到一个膜蛋白,它编码一个推测的G蛋白偶联受体(G protein-coupledreceptor, GPCR),据此我们将其命名为OsGPCR1,并对其进行深入研究。 OsGPCR1的cDNA全长为1407bp,编码468个氨基酸,在蛋白水平上的同源性比较结果显示,该基因与动物中研究的比较多的异源三聚体G蛋白偶联受体(G Protein- Coupled Receptor)同源性达到44%。经过跨膜结构域预测表明OsGPCR1具有9TMs结构,以GFP为标签的亚细胞定位表明OsGPCR1定位在膜上。GTP酶活性测定试验表明,OsGPCR1蛋白能够激活水稻RGA的GTP酶活性,此外,以泛素裂解体系为基础的酵母双杂交实验表明,OsGPCR1能够与RGA相互作用。说明OsGPCR1编码的蛋白是水稻中的一个G蛋白偶联受体。 OsGPCR1的表达受低温、干旱、高盐的诱导,但不受ABA,GA,ACC,IAA的诱导。在『F常生长条件下,OsGPCR1在水稻各器官中均有表达,但强弱有所不同。 在拟南芥和水稻中超表达OsGPCR1都能显著增强转基因植物对干旱、高盐、低温的耐受性。而在水稻中抑制OsGPCR1的表达,转基因水稻呈现出干旱、高盐、低温的敏感性。对转基因拟南芥下游基因的分析表明,超表达OsGPCR1能够在非胁迫条件下激活CBF途径中相关基因的表达。结合OsGPCR1不受ABA诱导的表达模式,我们推测OsGPCR1可能是通过不依赖于ABA这条途径而传递信号的。借助超表达和转反义水稻材料,利用水稻全基因组芯片研究OsGPCR1靶基因的结果表明,不论OsGPCR1基因表达量的降低或上升,都导致大约30%的与转运相关的基因的表达量发生改变。这暗示OsGPCR1可能通过囊泡运输传递胞外信号。此外FM4-64对超表达和转反义水稻幼根细胞染色标记后,OsGPCR1反义抑制水稻细胞内囊泡在细胞两端呈聚集状,即形成“BFA区间”,而超表达OsGPCRI水稻细胞内囊泡呈密集状。这些结果都表明OsGPCR1可能通过调控囊泡运输而将胞外胁迫信号传递进胞内。

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G蛋白参与了哺乳动物内多种细胞信号途径,但其在植物花粉萌发和花粉管发育过程中的细胞学定位、生化特性及功能研究比较滞后,有关这方面的研究报道较少。在显花植物授粉受精过程中,具顶端极性生长特性的花粉管是雄性生殖单位的载体,也是研究细胞生长分子调控机理的理想体系。与被子植物相比,裸子植物具有生长周期长,花粉管生长缓慢、易分叉等特点,具有不同于被子植物花粉发育的独特发育模式。对于裸子植物花粉萌发和花粉管生长的调控机理,目前尚不十分清楚。本文以松类植物中比较有代表性的裸子植物青杆(Piceawillsonii)和白皮松(Pinus bungeana)花粉为试材,应用免疫分析和间接免疫荧光显微镜技术,结合药理学实验和FTIR手段,研究了异三聚体G蛋白和小G蛋白在花粉管细胞中的定位、生化特性及其在花粉管发育中的调控作用。结果如下: 应用Western Blotting技术和来自于抗哺乳动物中不同序列G蛋白O【亚基抗体,我们在白皮松花粉管中检测到一条分子量为40 kDa左右的蛋白。去污剂处理显示,该蛋白与质膜偶联。间接免疫荧光显微镜实验发现,在花粉管发育的整个时期,代表Ga蛋白的荧光均一的分布在整个质膜区域,尤其在尖端皮层区域荧光最亮,显示此处该蛋白浓度最高。无论是在正常发育的花粉管抑或是发生弯曲或扭曲生长的花粉管,均呈现同样的分布模式。随着花粉管发育,Ga蛋白表达量发生变化。在花粉管发育中期,Ga蛋白表达量比较高;随着花粉管离体培养时间的延长,Ga蛋白表达量下降。另外,在花粉刚刚萌发时,Ga蛋白表达量也比较低。 对白皮松花粉萌发进行的药理学实验显示,G蛋白调节剂 CTX和PTX对白皮松花粉管的影响呈现双阶段效应。当添加的药剂浓度小于400 ng mL-I时,无论CTX还是PTX均抑制了花粉萌发和花粉管生长,且花粉管容易破裂;而当二者浓度分别升至500 ng mL-I时,同对照相比,花粉管生长明显受到促进。这一结果不支持Ma等人在百合花粉中的研究结果。进一步应用FTIR技术分析发现,当用浓度为400 ng mL-I CTX或PTX处理花粉管时,花粉管细胞壁酚类物质增加,而纤维素、半纤维素、木聚糖等物质下降,这可能是导致此浓度处理下花粉管易破裂的原因。这些结果显示了G蛋白a亚基参与了白皮松花粉管生长,CTX和PTX可能通过下游对其敏感的功能蛋白而非Ga本身,影响着花粉管生长并调控着花粉管壁的建成。 利用来源于烟草的抗NtRacl抗体和拟南芥的抗ROPs抗体,应用WeternBlotting技术,我们在青杆花粉管中检测到分子量为23kDa的多肽。间接免疫荧光显微镜实验显示,在花粉萌发18和24小时后,Rac蛋白主要定位于花粉管尖端质膜区域,时而会延伸到顶端两侧区域,但从尖端到基部存在浓度梯度,这种分布模式多在花粉管发育的后期观察到。Rac蛋白在青杆花粉管不同发育时期的分布模式变化可能和花粉管的生长状态有关,在花粉管发育早期和中期,正是花粉管旺盛生长期,Rac蛋白的尖端定位保证了花粉管的极性生长。对Rac蛋白在花粉管的分布进行的连续切片扫描发现,Rac蛋白不但分布在质膜上,并与质膜偶联,而且在胞质中亦有分布。通过对一系列正常发育(即极性生长的花粉管)和畸形发育的花粉管进行观察发现,Rac蛋白主要分布在旺盛生长的花粉管尖端质膜或离顶端20 Vm处,在分叉的生长缓慢的分枝端分布较少。而在那些发生分叉生长的花粉管中,处于次要位置的基本停止生长的分枝端几乎没有Rac蛋白存在。在顶端发生膨大的花粉管中,Rac蛋白均匀分布在花粉管整个质膜上,丧失浓度梯度,失去极性生长。这些结果显示了Rac蛋白参与了青杆花粉管生长。 应用抗NtRacl抗体进行的间接免疫荧光显微镜定位实验,我们在正在生长的花粉管的管核中观察到明亮的荧光,显示了有Rac蛋白的存在。当精细胞在花粉粒中未移动到花粉管中时,几乎没有观察到荧光信号。随着花粉管发育,两个精细胞的位置发生变化,当其中一个较大的精细胞移动到花粉管中时,观察到明亮的荧光信号,这些结果显示了Rac蛋白可能参与了管核或精细胞在花粉管内的移动。

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赤霉素(gibberellins,GAs)和油菜甾醇(brassinosteroids,BRs)在细胞伸长和植株形态建成等方面发挥重要的生理作用,但它们在分子水平上的相互作用仍然未知。在本实验室前期的芯片工作中筛选到受GA诱导表达的GAST家族基因OsGSR1(GA-stimulated gene in rice) (GenBank AY604180)。该基因全长cDNA为588 bp,编码110个氨基酸,OsGSR1具有GAST家族成员的共同特点。OsGSR1基因的表达受GA3诱导,同时受PAC抑制。基因表达模式分析表明OsGSR1在水稻的根、茎、幼穗和小花等多种组织和器官中表达。前期工作已获得转基因水稻。 本论文研究表明,OsGSR1 RNAi转基因水稻表现为初生根缩短、叶片直立、节间缩短和结实率降低等与GA和BR相关的表型。OsGSR1 RNAi转基因水稻对外源GA3敏感性降低,Real-time PCR分析表明在OsGSR1 RNAi转基因水稻中OsGA20ox2和SLR1的转录水平增高,GC-MS分析显示内源GA4含量增高,这些结果说明转基因材料中GA信号削弱。因此,OsGSR1是GA信号途径的正调控因子。另一方面,实验证据表明,外源BL处理可以抑制OsGSR1基因的表达,OsGSR1 RNAi转基因水稻不但可以响应外源BL处理,并且在叶夹角实验中显现出对外源BL更加敏感的特性。在OsGSR1 RNAi转基因水稻中,BR受体基因OsBRI1与合成基因OsDWARF表达量上调。外源添加BL可以恢复OsGSR1 RNAi转基因水稻矮化表型,上述结果说明OsGSR1可能作用于BR生物合成途径。酵母双杂交筛选、体外Pull-down结果和体内BiFC实验都证实OsGSR1可以与DIM/DWF1互作。在BR生物合成途径中,DIM/DWF1催化从24-亚甲基固醇(24-methylenecholesterol)到油菜甾醇(campesterol)的转化。GC-MS测定内源BRs含量结果进一步证实,转基因水稻中DIM/DWF1催化反应产物积累量减少,说明该反应受到明显抑制。所以,OsGSR1是通过直接作用于BR合成酶来调控BR生物合成。 综上所述,OsGSR1是GA信号途径的正调控因子,并且OsGSR1通过调节SLR1的表达参与到GA信号转导途径。OsGSR1和DIM/DWF1的互作说明OsGSR1直接参与了BR的生物合成过程。因此,我们的实验证明OsGSR1介导了GA和BR这两条激素信号转导途径的相互作用,从而调节了水稻植株的生长发育。

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非生物逆境胁迫严重影响植物的生长发育和作物的产量。解析逆境下植物体基 因表达及信号传导网络调控机制对于阐明植物的适应性反应及培育耐逆性作物具有 重要理论和现实意义。 雨生红球藻(Haematoccoccus pluvialis)通过累积虾青素来适应胁迫环境,茉莉 酮酸甲酯(methyl jasmonate,MJ)和赤霉素(gibberellins,GA)对植物胁迫适应过 程中相关基因的诱导表达起到重要的调控作用。本文选取 H. pluvialis 作为出发株, 研究了 MJ 和 GA 对虾青素合成途径关键酶 β-胡萝卜素酮化酶基因的调控作用。 在其中一种 H. pluvialis 中克隆获得了三种不同的 β-胡萝卜素酮化酶基因。通过基 因组步移,获得了三种 β-胡萝卜素酮化酶的5’ 侧翼序列(5’-flanking region),其 中存在着多样的顺式作用元件,包括 MJ 和 GA 的顺式作用位点。进一步的实验 表明,MJ 和 GA 的诱导处理,可以促进 H. pluvialis 中虾青素的累积;同时,MJ 和 GA 调控了三种 β-胡萝卜素酮化酶基因的转录水平。 南极小球藻(Chlorella vulgaris NJ-7)可以在较大温度和盐度变化范围内存活, 是研究生物胁迫适应机理的理想模式生物。对数期的 C. vulgaris NJ-7 主要的脂肪酸

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A 10-fold BAC library for giant panda was constructed and nine BACs were selected to generate finish sequences. These BACs could be used as a validation resource for the de novo assembly accuracy of the whole genome shotgun sequencing reads of giant panda newly generated by the Illumina GA sequencing technology. Complete sanger sequencing, assembly, annotation and comparative analysis were carried out on the selected BACs of a joint length 878 kb. Homologue search and de novo prediction methods were used to annotate genes and repeats. Twelve protein coding genes were predicted, seven of which could be functionally annotated. The seven genes have an average gene size of about 41 kb, an average coding size of about 1.2 kb and an average exon number of 6 per gene. Besides, seven tRNA genes were found. About 27 percent of the BAC sequence is composed of repeats. A phylogenetic tree was constructed using neighbor-join algorithm across five species, including giant panda, human, dog, cat and mouse, which reconfirms dog as the most related species to giant panda. Our results provide detailed sequence and structure information for new genes and repeats of giant panda, which will be helpful for further studies on the giant panda.

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Environmental. factors that affect the growth and microcystin production of microcystis have received worldwide attention because of the hazards microcystin poses to environmental safety and public health. Nevertheless, the effects of organic anthropogenic pollution on microcystis are rarely discussed. Gibberellin A(3) (GA(3)) is a vegetable hormone widely used in agriculture and horticulture that can contaminate water as an anthropogenic pollutant. Because of its common occurrence, we studied the effects of GA3 on growth and microcystin production of Microcystis aeruginosa (M. aeruginosa) PCC7806 with different concentrations (0.001-25mg/L) in batch culture. The control was obtained without gibberellin under the same culture conditions. Growth, estimated by dry weight and cell number, increased after the GA3 treatment. GA3 increased the amounts of chlorophyll a, phycocyanin and cellular-soluble protein in the cells of M. aeruginosa PCC7806, but decreased the accumulation of water-soluble carbohydrates. In addition, GA3 was observed to affect nitrogen absorption of the test algae, but to have no effect on the absorption of phosphorus. The amount of microcystin measured by enzyme-Linked immunosorbent assay (ELISA) increased in GA3 treatment groups, but the stimulatory effects were different in different culture phases. It is suggested that GA3 increases M. aeruginosa growth by stimulating its absorbance of nitrogen and increasing its ability to use carbohydrates, accordingly increasing cellular pigments and thus finally inducing accumulation of protein and microcystin. (C) 2007 Elsevier GmbH. All rights reserved.

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Intron loss and its evolutionary significance have been noted in Drosophila. The current study provides another example of intron loss within a single-copy Dfak gene in Drosophila. By using polymerase chain reaction (PCR), we amplified about 1.3 kb fragment spanning intron 5-10, located in the position of Tyr kinase (TyK) domain of Dfak gene from Drosophila melanogaster species group, and observed size difference among the amplified DNA fragments from different species. Further sequencing analysis revealed that D. melanogaster and D. simulans deleted an about 60 bp of DNA fragment relative to other 7 Drosophila species, such as D. elegans, D. ficusphila, D. biarmipes, D. takahashii, D. jambulina, D. prostipennis and D. pseudoobscura, and the deleted fragment located precisely in the position of one intron. The data suggested that intron loss might have occurred in the Dfak gene evolutionary process of D. melanogaster and D. simulans of Drosophila melanogaster species group. In addition, the constructed phylogenetic tree based on the Dfak TyK domains clearly revealed the evolutionary relationships between subgroups of Drosophila melanogaster species group, and the intron loss identified from D. melanogaster and D. simulans provides a unique diagnostic tool for taxonomic classification of the melanogaster subgroup from other group of genus Drosophila.

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The structural and magnetic properties of Cu+ ions-implanted GaN films have been reported. Eighty kilo-electron-volt Cu+ ions were implanted into n-type GaN film at room temperature with fluences ranging from 1 x 10(16) to 8 x 10(16) cm(-2) and subsequently annealed at 800 degrees C for 1 h in N-2 ambient. PIXE was employed to determine the Cu-implanted content. The magnetic property was measured by the Quantum Design MPMS SQUID magnetometer. No secondary phases or clusters were detected within the sensitivity of XRD. Raman spectrum measurement showed that the Cu ions incorporated into the crystal lattice positions of GaN through substitution of Ga atoms. Apparent ferromagnetic hysteresis loops measured at 10 K were presented. The experimental result showed that the ferromagnetic signal strongly increased with Cu-implanted fluence from 1 x 10(16) to 8 x 10(16) cm(-2).

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Atomic configurations and formation energies of native defects in an unsaturated GaN nanowire grown along the [001] direction and with (100) lateral facets are studied using large-scale ab initio calculation. Cation and anion vacancies, antisites, and interstitials in the neutral charge state are all considered. The configurations of these defects in the core region and outermost surface region of the nanowire are different. The atomic configurations of the defects in the core region are same as those in the bulk GaN, and the formation energy is large. The defects at the surface show different atomic configurations with low formation energy. Starting from a Ga vacancy at the edge of the side plane of the nanowire, a N-N split interstitial is formed after relaxation. As a N site is replaced by a Ga atom in the suboutermost layer, the Ga atom will be expelled out of the outermost layers and leaves a vacancy at the original N site. The Ga interstitial at the outmost surface will diffuse out by interstitialcy mechanism. For all the tested cases N-N split interstitials are easily formed with low formation energy in the nanowires, indicating N-2 molecular will appear in the GaN nanowire, which agrees well with experimental findings.

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一种使用生长半绝缘砷化镓的石英管在砷化镓中掺碳的方法,包括如下步骤:步骤1:将7N Ga和7N As进行多晶合成,形成GaAs多晶;步骤2:将合成好的GaAs多晶、籽晶和B2O3放入PBN坩埚;步骤3:将PBN坩埚放入石英管的石英体中;步骤4:将纯石墨固定在石英管的石英帽上的石英槽内;步骤5:将石英体和石英帽盖合,抽真空,用氢氧焰焊接石英管的石英体和石英帽;步骤6:将焊接后的石英管放入VGF单晶炉,进行气氛掺杂,单晶生长;步骤7:将晶体生长后的PBN坩埚放入甲醇内浸泡,得到GaAs单晶,完成GaAs单晶的制备。

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We studied the structural and optical properties of high Al-content AlInGaN epilayers with different thicknesses grown on GaN/sapphire templates by metalorganic chemical vapor deposition (MOCVD). Direct evidences of the gradual evolution of the content of Al, Ga and In along the growth direction were obtained. When the film thickness was over a certain value, however, the AlInGaN epilayer with constant element contents began to form. These results were also supported by the blue shift and splitting of the photoluminescence (PL) peak. For the thinnest epilayer, the surface was featured with outcrops of threading dislocations (TDs) which suggested a spiral growth mode. With increase in thickness, step-flow growth mode and V-shaped pits were observed, and the steps terminated at the pits. (C) 2008 Elsevier B. V. All rights reserved.

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InGaN/GaN multi-quantum-well blue (461 +/- 4 nm) light emitting diodes with higher electroluminescence intensity are obtained by postgrowth thermal annealing at 720 C in O-2-ambient. Based on our first-principle total-energy calculations, we conclude that besides dissociating the Mg-H complex by forming H2O, annealing in O-2 has another positive effect on the activation of acceptor Mg in GaN. Mg can be further activated by the formation of an impurity band above the valence band maximum of host GaN from the passivated Mg-Ga-O-N complex. Our calculated ionization energy for acceptor Mg in the passivated system is about 30 meV shallower than that in pure GaN, in good agreement with previous experimental measurement. Our model can explain that the enhanced electroluminescence intensity of InGaN/GaN MQWs based on Mg-doped p-type GaN is due to a decrease in the ionization energy of Mg acceptor with the presence of oxygen. (C) 2008 American Institute of Physics.

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This paper presents a study of the transformation of high-temperature AlN (HT-AlN) interlayer (IL) and its effect on the strain relaxation of Al0.25Ga0.75N/HT-AlN/GaN. The HT-AlN IL capped with Al0.25Ga0.75N transforms into AlGaN IL in which the Al composition increases with the HT-AlN IL thickness while the total Ga content keeps nearly constant. During the HT-AlN IL growth on GaN, the tensile stress is relieved through the formation of V trenches. The filling up of the V trenches by the subsequent Al0.25Ga0.75N growth is identified as the Ga source for the IL transformation, whose effect is very different from a direct growth of HT-AlGaN IL. The a-type dislocations generated during the advancement of V trenches and their filling up propagate into the Al0.25Ga0.75N overlayer. The a-type dislocation density increases dramatically with the IL thickness, which greatly enhances the strain relaxation of Al0.25Ga0.75N. (c) 2008 American Institute of Physics.

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Using the first-principles methods, we study the electronic structure, intrinsic and extrinsic defects doping in transparent conducting oxides CuGaO2. Intrinsic defects, acceptor-type and donor-type extrinsic defects in their relevant charge state are considered. The calculation result show that copper vacancy and oxygen interstitial are the relevant defects in CuGaO2. In addition, copper vacancy is the most efficient acceptor. Substituting Be for Ga is the prominent acceptor, and substituting Ca for Cu is the prominent donors in CuGaO2. Our calculation results are expected to be a guide for preparing n-type and p-type materials in CuGaO2.