Beam quality improvement of laser diode array by using off-axis external cavity

A novel off-axis external cavity is designed for laser diode array to improve the beam quality. In this external cavity, a circle aperture with variable size is used as a spatial filter. The diameter of aperture is optimized to 1.2mm and the off-axis angle of external cavity is optimized at 2.6 deg. In the optimal case, the beam parameter product (BPP) of laser diode array is reduced to 121 mm. mrad from 1050 mm. mrad with external cavity optical efficiency of 81%. (C) 2007 Optical Society of America.

Quality improvement of GaInNAs/GaAs quantum wells grown by plasma-assisted molecular beam epitaxy

The optimum growth condition of GaInNAs/GaAs quantum wells (QWs) by plasma-assisted molecular beam epitaxy was investigated. High-resolution X-ray diffraction and photoluminescence (PL) measurements showed that ion damage drastically degraded the quality of GaNAs and GaInNAs QWs and that ion removal magnets can effectively remove the excess ion damage. Remarkable improvement of PL intensity and obvious appearance of pendellosung fringes were observed by removing the N ions produced in the plasma cell. When the growth rate increased from 0.73 to 1.2 ML/s, the optimum growth temperature was raised from 460 degreesC to 480 degreesC and PL peak intensity increased two times. Although the N composition decreased with increasing growth rate, degradation of optical properties of GaInNAs QWs was observed when the growth rate was over 0.92 ML/s. Due to low-temperature growth of GaInNAs QWs, a distinctive reflection high-energy electron diffraction pattern was observed only when the GaAs barrier was grown under lower As-4 pressure. The samples with GaAs barriers grown under lower As-4 pressure (V/III ratio about 24) exhibited seven times increase in PL peak intensity compared with those grown under higher As-4 pressure (V/III ratio about 50). (C) 2001 Elsevier Science B,V. All rights reserved.

Quality improvement of GaInNAs/GaAs quantum wells grown by plasma-assisted molecular beam epitaxy

The optimum growth condition of GaInNAs/GaAs quantum wells (QWs) by plasma-assisted molecular beam epitaxy was investigated. High-resolution X-ray diffraction and photoluminescence (PL) measurements showed that ion damage drastically degraded the quality of GaNAs and GaInNAs QWs and that ion removal magnets can effectively remove the excess ion damage. Remarkable improvement of PL intensity and obvious appearance of pendellosung fringes were observed by removing the N ions produced in the plasma cell. When the growth rate increased from 0.73 to 1.2 ML/s, the optimum growth temperature was raised from 460 degreesC to 480 degreesC and PL peak intensity increased two times. Although the N composition decreased with increasing growth rate, degradation of optical properties of GaInNAs QWs was observed when the growth rate was over 0.92 ML/s. Due to low-temperature growth of GaInNAs QWs, a distinctive reflection high-energy electron diffraction pattern was observed only when the GaAs barrier was grown under lower As-4 pressure. The samples with GaAs barriers grown under lower As-4 pressure (V/III ratio about 24) exhibited seven times increase in PL peak intensity compared with those grown under higher As-4 pressure (V/III ratio about 50). (C) 2001 Elsevier Science B,V. All rights reserved.

The restoration of aquatic macrophytes for improving water quality in a hypertrophic shallow lake in Hubei Province, China

This paper deals with a case study of the restoration of submerged macrophytes for improving water quality in a hypertrophic shallow lake, Lake Donghu of Wuhan, Hubei Province, China. Macrophyte restoration experiments were conducted in large-scale enclosures established in three sublakes of different trophic status, and the effectiveness for water quality improvement was tested by using the enclosure experiment in the hypertrophic sublake. Water quality was remarkably improved after the reestablishment of aquatic macrophytes. It is suggested that the submerged vegetation of less polluted sublakes could be capable of recovering spontaneously once the stocking of herbivorous fishes has been ceased, and the K-selected plants such as Potamogeton maackianus should be introduced into these sublakes to enhance the stability of aquatic vegetation. However, it may not be possible and economical to restore the submerged macrophytes in severely polluted basins unless external pollution has been cut off and internal nutrient loadings considerably reduced. In this case, the r-selected submerged plants should be used as the pioneer species for macrophyte recovery. (C) 2001 Elsevier Science B.V. All rights reserved.

Recent research results on deep level defects in semi-insulating InP - Application to improve material quality

An apparent defect suppression effect has been observed in InP through an investigation of deep level defects in different semi-insulating (SI) InP materials. Quality improvement of SI-InP based on the defect suppression mechanism is presented.

植物油脂积累机理和品质改良—利用RNAi改良大豆油脂品质—四合木茎积累三脂酰甘油特征

利用RNAi改良大豆油脂品质 大豆[Glycine max (L.) Merr.]起源于中国，栽培历史悠久，是重要的粮食作物， 同时也是植物油和蛋白的重要来源。随着经济的发展和生活水平的提高，人们不但对大豆的需求量大大增加，同时对大豆的品质也提出了更高的要求。近年来，我国大豆进口量逐年攀升，已远远超过本国生产量。国外转抗除草剂转基因大豆大面积种植大大降低了生产成本，直接影响了我国大豆生产。因此，提高产量和改良品质是当前中国大豆生产所面临的重要课题。基因工程是大豆品种改良更为有效和快速的方法，但是由于历史原因我国的大豆转基因育种与发达国家尚存在一定差距，对我国的大豆生产贡献十分有限。因此，建立高效的大豆转化体系，加强大豆基因工程研究和育种是解决大豆面临困境的关键。 本研究的目的是以我国主要栽培大豆品种（黑农、合丰和东农等）为材料，利用GUS（β-glucuronidase）报告基因和RNAi技术，建立高效的大豆基因转化体系和基因功能研究体系。为大豆产量和品质基因工程改良提供技术手段和理论基础。结果如下： 以大豆下胚轴为外植体，对分生组织产生不定芽的频率进行了研究。培养基中添加高浓度BAP（6-benzylaminopurine）可以诱导外植体分生组织增殖产生不定芽的发生率;在培养基中添加银离子可以明显地促进大豆单个外植体多芽的产生，使得诱导不定芽总数目显著增加;不同基因型大豆再生不定芽能力有着较大区别，黑农44，黑农37，合丰35，合丰39等品种再生能力强;相对于大豆子叶节等再生系统，大豆下胚轴体系具有高效高频的再生特点（总的再生频率高于80%），且重复性好，容易操作。 以大豆下胚轴为外植体，用含有GUS报告基因的根癌农杆菌对其进行遗传转化，并重点对农杆菌菌液浓度、农杆菌侵染时间、乙酰丁香酮（AS）和抗氧化剂浓度等因素对农杆菌大豆转化效率的影响进行了研究。组织化学染色结果显示GUS基因在外植体顶端表达强烈，表达位置主要位于初生芽基部周围的分生组织。 农杆菌浸染时间以 4h 为最佳，此时的GUS瞬时表达频率可达73.0%;培养基中添加浓度为200μmol/L的乙酰丁香酮，可以显著增加GUS瞬时表达频率。抗氧化剂可以显著降低共培养阶段外植体的褐化和坏死率，进而显著提高农杆菌转化效率。用根癌农杆菌转化大豆下胚轴的方法得到了表达GUS基因转基因大豆株系。 利用大豆油酸去饱和酶基因(FAD2-1;Genbank, L43920)在第315-852碱基之间的基因片断构建了反向重复的RNAi表达载体，以农杆菌介导大豆下胚轴转化方法进行转化，并且获得转基因植株。经过PCR，Southern杂交和转基因后代的脂肪酸分析，表明沉默结构已经成功整合到大豆基因组中，并成功抑制了内源基因的表达。与栽培大豆品种相比较，转基因大豆种子的脂肪酸组成发生显著变化，油酸含量由栽培大豆的18.1％增加到71.5％¬-81.9％;亚油酸含量从栽培大豆的46.4％降到了约3.4％。 栽培大豆种子中油酸去饱和比率（ODP, oleic desaturation proportion）为0.76 到 0.84，转基因大豆种子的油酸去饱和比率降为0.06-0.26，表明Δ12-去饱和酶活性降低了74%-94%。上述结果表明，我们构建的RNAi反向重复序列沉默结构高效地抑制了大豆种子FAD2-1基因。 在本研究中，我们通过外源GUS基因的表达和内源FAD2基因的抑制，成功地建立了以大豆下胚轴为外植体的高效农杆菌介导大豆转化体系，并获得了相应的转基因株系。本研究对我国大豆品种基因工程改良以及进一步大豆功能基因组研究有重要参考价值。 四合木茎积累三脂酰甘油特征 四合木(Tetraena mongotica Maxim)是蒺藜科(Zygophyllaceac)四合木属唯一的种，是地球上最具代表性的古老残遗濒危珍稀植物。由于四合木极易燃烧，当地居民称其为“油柴”。 通过对四合木内可能存在的“油”成分进行了分析，我们发现其茎组织含有大量的三脂酰甘油（Triacylglycerols），含量达到46 mg/g DM。在韧皮部中更高，达到90 mg/g DM。我们通过半薄切片对四合木中三脂酰甘油在不同组织的分布和存在形式进行了研究，发现三脂酰甘油主要以油体形式存在于木质部和韧皮部的薄壁组织中。在韧皮部中，几乎所有的薄壁细胞都含有大量的油体。 三脂酰甘油在植物的生长发育中起着非常重要的作用。作为植物生长发育所需的碳源和能量，三脂酰甘油一般储存在植物的种子和果实中。虽然也有关于其在茎和叶中发现的报道，但是含量很少。四合木茎组织含有大量的三脂酰甘油，这种现象可能与四合木茎中存在茎特异油脂合成酶系统有关。因此，克隆相关基因并在作物中表达，将对能源植物的开发具有重要意义。

Effects of reactor pressure on GaN nucleation layers and subsequent GaN epilayers grown on sapphire substrate

The influence of reactor pressure on GaN nucleation layer (NL) and the quality of subsequent GaN on sapphire is studied. The layers were grown by low-pressure metalorganic chemical vapor deposition (MOCVD) on c-plane sapphire substrates and investigated by in situ laser reflectometry, atomic force microscope, scanning electron microscope, X-ray diffraction and photoluminescence. With the increase of reactor pressure prior to high-temperature GaN growth, the size of GaN nuclei formed after annealing decreases, the spacing between nucleation sites increases and the coalescence of GaN nuclei is deferred. The optical and crystalline qualities of GaN epilayer were improved when NLs were deposited at high pressure. The elongated lateral overgrowth of GaN islands is responsible for the quality improvement. (C) 2003 Elsevier Science B.V. All rights reserved.

Effect of rapid thermal annealing on GaInNAs/GaAs quantum wells grown by plasma-assisted molecular-beam epitaxy

We have studied the effect of rapid thermal annealing (RTA) on GaInNAs/GaAs quantum wells (QWs) grown by molecular-beam epitaxy using a dc plasma as the N source. It was found that RTA at low temperature (LT, 650 degrees C) and high temperature (HT, 900 degrees C) could both improve the QW quality significantly. To clarify the mechanism of quality improvement by RTA, a magnetic field perpendicular to the path of the N plasma flux was applied during the growth of the GaInNAs layers for the sake of comparison. It was found that LT-RTA mainly removed dislocations at interfaces related to the ion bombardment, whereas, HT-RTA further removed dislocations originating from the growth. LT-RTA caused only a slight blueshift of photoluminescence peak wavelength, probably due to defect-assisted interdiffusion of In-Ga at the QW interfaces. The blueshift caused by HT-RTA, on the other hand, was much larger. It is suggested that this is due to the fast defect-assisted diffusion of N-As at the QW interfaces. As defects are removed by annealing, the diffusion of In-Ga at interfaces would be predominant. (C) 2000 American Institute of Physics. [S0003- 6951(00)01535-7].

1.3 mu m GaInNAs/GaAs quantum well lasers and photodetectors

The growth of GalnNAs/GaAs quantum well (QW) has been investigated by solid-source molecular beam epitaxy (MBE). N was introduced by a dc-active plasma source. Highest N concentration of 2.6% in GaInNAs/GaAs QW was obtained, corresponding to the photoluminescence peak wavelength of 1.57 mum at 10K. The nitrogen incorporation behavior in MBE growth and the quality improvement of the QW have been studied in detail. 1.3 mum GaInNAs/GaAs SQW laser and MQW resonant-cavity enhanced photodetector have been achieved.

鞑靼荞麦（Fagopyrum tataricum Gaertn.）离体再生与遗传转化研究

鞑靼荞麦是我国特有的农业产品，具有抗寒耐旱特性和较高的营养保健功能。荞麦的开花习性及遗传特点导致其人工杂交授粉难以成功，这成为荞麦杂交育种难以获得突破的重要原因。因此利用转基因技术导入有益基因有可能成为荞麦遗传改良的新途径，而再生及转化体系的建立是开展转基因研究的基础。 本文研究了苗龄、外植体、几种激素配比对鞑靼荞麦（Fagopyrum tataricum Gaertn.）离体培养的影响，初步建立了鞑靼荞麦离体再生体系。结果表明，鞑靼荞麦离体再生的最佳取材时间为苗龄6-8d；诱导愈伤组织的最适培养基为MS＋2.0 mg/L 2,4-D＋1.5 mg/L 6-BA，子叶诱愈率达75％左右，下胚轴的可高达86.62％；愈伤组织分化的最适培养基为MS 0.1mg/L IAA＋2.0mg/L 6-BA＋1.0 mg/L KT＋0.5mg/L TDZ，下胚轴的分化率可达9.52%。下胚轴的诱愈率与分化率均高于子叶，更适于离体再生培养。培养基中加入AgNO3后，能有效降低褐化率。生根最适培养基为含有0.5mg/L NAA的1/2MS培养基，生根率在50％左右。TDZ在诱导鞑靼荞麦的愈伤组织分化出芽的过程中起到明显的促进作用，可提高分化率约20%。 在上述研究基础上，本文还对鞑靼荞麦的遗传转化体系进行了探索性研究。分别利用根癌农杆菌（Agrobacterium tumefaciens）介导法和微粒轰击法（基因枪法）对黑水苦荞下胚轴进行遗传转化。 在农杆菌介导的方法中，携带有质粒pCAMBIA2301的农杆菌菌株EHA105用于转化。载体质粒pCAMBIA2301包含有gus和npt-II 基因, 并受35s启动子驱动。研究结果表明，在侵染方式选择上，浸泡方式比吸打方式更有效，根癌农杆菌侵染的较适浓度为OD600＝0.5，共培养3天，恢复培养7天，能检测到gus基因的表达。 基因枪法使用质粒pBI121，同样包含有gus和npt-II基因, 并受CaMV35s 启动子驱动。轰击距离为9cm较合适，甘露醇前处理在本研究中未表现出明显优势。 两种转化方法比较，基因枪法比农杆菌介导法更快速有效。 本研究为进一步的遗传操作研究打下基础。 Tartary buckwheat (Fagopyrum tataricum Gaertn.), the traditional and unique agricultural product of China, is a kind of crop with strong drought and cold tolerance, abundant nutrition and high medical value. Artificial hybridization is hard in buckwheat because of its flowering habits and genetic characteristics, which leads to no breakthrough in tartary buckwheat breeding. However, biotechnological approaches, especially genetic transformation for the direct introduction of good genes into tartary buckwheat for quality improvement, hold great promise. In this study, we established tartary buckwheat regeneration system in vitro. It is the foundation for genetic manipulation of this crop. The effects of seedling age, hypocotyl and cotyledon as explants, and proportions of several growth regulators were tested in tissue culture of tartary buckwheat for establishing its in vitro regeneration system. The results showed that the best seedling age for callus induction was 6 to 8 days. On the MS medium containing 2.0mg/L 2, 4-D and 1.5mg/L 6-BA, the induction rate of callus from hypocotyls was up to 86.62%, while from cotyledons was about 75％. The suitable shooting medium was the MS medium+0.1mg/L IAA+2.0mg/L 6-BA+1.0 mg/L KT+0.5mg/L TDZ, and the shooting rate from hypocotyls was 9.52%. The callus induction and shooting rates were higher from hypocotyls than from cotyledons. Browning reduced when the medium mixed with AgNO3. Half strength MS supplemented with 0.5mg/L NAA was the best for rooting, the rate was around 50% after 30 days culture. TDZ can accelerate the shoot differentiation distinctively, and it could improve the shooting rate nearly 20%. On the base of above, the explorative research of the genetic transformation in tartary buckwheat was done. In the study, hypocotyls from Heishui tartary buckwheat were transformed by Agrobacterium-mediated method and microprojectile bombardment method (gene-gun), comparatively. In Agrobacterium-mediated method, a disarmed Agrobacterium tumefaciens strain EHA105 harboring plasmid pCAMBIA2301 was used. The vector pCAMBIA2301 contains gus and npt-II genes, driven by CaMV35s promoter. The results showed that the appropriate concentration of Agrobacterium tumefaciens for infecting was OD600＝0.5, and co-culture time was 3d. Seven days later after coculture, GUS expression could be tested. In particle bombardment transformation, plasmid pBI121 was used. pBI121 also contains gus and npt-II genes, driven by 35s promoter. Hypocotyls pretreated with mannitol, no effect was observed, and the suitable distance of bombardment is 9cm. Comparing with Agrobacterium-mediated method, gene-gun method is more convenient and effective. All above results could be a basic work for further study in tartary buckwheat transformation.

青稞B组醇溶蛋白基因与上游调控区的克隆及基因表达

高等植物种子胚乳贮藏蛋白是种子发芽时的主要氮源，也是人类和动物食用植物蛋白的主要来源。大麦种子胚乳贮藏蛋白主要是醇溶蛋白（hordeins），占大麦胚乳总蛋白的50–60%。根据大麦醇溶蛋白的大小和组成特点，大麦醇溶蛋白被划分为三种类型：富硫蛋白亚类(B，γ-hordeins)、贫硫蛋白亚类（C-hordeins)以及高分子量蛋白亚类(D-hordeins)。B组和C组醇溶蛋白是大麦胚乳的两类主要贮藏蛋白，它们分别占大麦总醇溶蛋白成分的70–80%和10–12%。遗传分析表明，大麦B、C、D和γ-组醇溶蛋白分别是由位于大麦第五染色体1H（5）上的Hor2、Hor1、Hor3和Hor5位点编码。Hor2位点编码大量分子量相同但组成不同的B组醇溶蛋白（B-hordein）。B-hordein的种类、数量和分布是影响大麦酿造、食用及饲养品质的重要因素之一。为深入了解B-hordein基因家族的结构和染色体组织，探明Hor2位点基因表达的发育调控机制，最终达到改良禾谷类作物籽粒品质的目的，本研究以青藏高原青稞为材料，采用同源克隆法，分别克隆B-hordein基因和启动子，通过原核生物表达验证B-hordein基因功能，并利用实时定量PCR探索B-hordein基因表达时空关系，取得如下研究结果： 1. 以具有特殊B组醇溶蛋白亚基组成的9份青藏高原青稞为材料，根据GenBank中三个B-hordein基因序列（GenBank No. X03103, X53690和X53691）设计一对引物，通过PCR扩增，获得23个B-hordein基因克隆并对其进行了序列分析。核苷酸序列分析表明，所有克隆均包含完整的开放阅读框。有11个克隆都存在一个框内终止密码子，推测这11个克隆可能是假基因。推测的氨基酸序列分析表明，所有大麦B-hordein具有相似的蛋白质基本结构，均包括一个高度保守的信号肽、中间重复区以及C-端结构域。不同大麦种重复区内重复基元的数目有较大差异。青稞材料Z07–2和Z26的B-hordeins仅具有12个重复基元结构，更接近于野生大麦。这些重复基元数目的差异导致了重复区序列长度和结构的变异。这种现象极可能是由于醇溶谷蛋白基因在进化过程中染色体的不平衡交换或复制滑动所造成的。对所克隆基因和禾本科代表性醇溶谷蛋白基因进行聚类分析，结果表明所有来自栽培大麦的B-hordeins聚类成一个亚家族，来自野生大麦的B-hordeins以及普通小麦的LMW-GS聚类成另外一个亚家族，表明这两个亚家族的成员存在显著差异。此外，我们发现B-hordein基因推测的C-末端序列具有一些有规律的特征：即具有相同C-末端序列的B-hordein基因在系统发生树中聚类为同一个亚组（除BXQ053，BZ09-1，BZ26-5分别单独聚为一类外）。这个特征将有助于我们对所有B组醇溶蛋白基因家族成员进行分类，避免了在SDS-PAGE电泳图谱上仅依靠大小分类的局限性。 2. 根据上述克隆的青稞B-hordein基因的5’端序列设计三条基因特异的反向引物，以青稞Z09和Z26的基因组DNA为模板，采用SON-PCR和TAIL-PCR技术分离克隆出8个B-hordein基因的上游调控序列（命名为Z09P和Z26P）。序列分析表明，推测的TATA box位于–80 bp，CAAT–like box位于–140 bp处。此外，Z09P和Z26P中有六个序列在–300 bp处均存在一个由高度保守的EM基序和类GCN4基序构成的胚乳盒(Endosperm Box，EB)，在约–560 bp处存在一个胚乳盒类似结构。而Z09P-2和Z26P-3不存在保守的胚乳盒或其类似结构，预示着这两个启动子所调控的基因表达可能受不同类型反式作用因子的调节，推测该启动子对基因的表达调控具有多样性。 3. 将B-hordein基因的开放阅读框定向克隆到表达载体pET-30a中，将其导入大肠杆菌表达菌株BL21中进行外源基因的诱导表达以验证所克隆基因的功能。结果表明仅含重组子pET-BZ07-2和pET-BZ26-5的BL21细菌有目的表达蛋白产生。在诱导3 h时的蛋白表达量最高；3 mM IPTG诱导的蛋白表达量要高于1 mM IPTG诱导的表达量。这为分离纯化B-hordein蛋白以及进一步研究其对大麦籽粒品质的影响奠定基础。 4. 根据从青稞Z09和Z26中分离克隆的B-hordein基因序列设计一对基因特异的引物，同时，选择大麦α-微管蛋白基因（GenBank no. U40042）为看家基因并设计特异引物，利用实时荧光定量PCR检测了青稞籽粒4个胚乳发育时间段的B-hordein基因表达，荧光定量结果显示：两份材料中B-hordein基因的表达量均随发育过程的进行而逐渐升高。Z09中B-hordein基因在开花后7天开始转录，而Z26开花4天后就有低水平B-hordein的表达，这表明Z26中B-hordein基因可能比Z09表达的较早或者Z09中B-hordein基因表达水平较低以致于不能被检测到。此外，在4个不同的胚乳发育时期中，Z26中B-hordein基因的表达量均高于Z09材料。在开花12天到18天的过程中，Z09和Z26中B-hordein基因的表达水平有一个急剧性的升高。这说明在不同胚乳发育时期，Hor2位点的B-hordein等位基因变异体存在mRNA的差异表达。 Seed endosperm storage proteins in higher plants are the main resources of nitrogen for germinating and plant proteins for human and animals. Barley prolamins (also called hordeins) are the major storage proteins in the endosperm and account for 50–60% of total proteins. Hordeins are classically divided into three groups: sulphur-rich (B, γ-hordeins), sulphur-poor (C-hordeins) and high molecular weight (HMW, D-hordeins) hordeins based on the size and composition. B-hordeins and C-hordeins are two major groups and each respectively account for about 70-80% and 10-12% of the total hordein fraction in barley endosperm. Genetic analysis showed that B-, C-, C-, γ-hordeins are encoded by Hor2, Hor1, Hor3 and Hor5 locus on the chromosome 1H (5). Hor2 locus is rich in alleles that encode numerous heterogeneous B-hordein polypeptides. It is reported that B-hordein species, quantity and distribution are significant factors affecting malting, food and feed quality of barley. To understand comprehensively the structure and organization of B-hordein gene family in hull-less barley and explore the developmental control mechanisms of Hor2 locus gene expression and eventually to better exploitation in crop grain quality improvement, we isolated and cloned B-hordein genes and promotors of hull-less barley from Qinghai-Tibet Plateau by PCR, and testified their expression founction in bacteria expression system and explore their spatial and temporal expression pattern by quantitative real time PCR. Our results are as followed, 1. Twenty-three copies of B-hordein gene were cloned from nine hull-less barley cultivars of Qinghai-Tibet Plateau with special B-hordein subunits and molecularly characterized by PCR, based on three B-hordein genes published previously (GenBank No. X03103, X53690 and X53691). DNA sequences analyses confirmed that the six clones all contained a full-length coding region of the barley B-hordein genes. Eleven clones all contain an in-frame stop codon and they are probably pseudogenes. The analysis of deduced amino acid sequences of the genes shows that they have similar structures including signal peptide domain, central repetitive domain, and C-terminal domain. The number of the repeats was largerly variable and resulted in polypeptides in different sizes or structures among the genes. Twelve such repeated motifs were found in Z07–2 and Z26, and they are close to those of the wild barleys, and it is most probably caused by unequal crossing-over and/or slippage during replication as suggested for the evolution of other prolamins. The relatedness of prolamin genes of barley and wheat was assessed in the phylogenetic tree based on their polypeptides comparison. Our phylogenetic analysis suggested that the predicted B-hordeins of cultivated barley formed a subfamily, while the B-hordeins of wild barleys and the two most similar sequences of LMW-GS of T. aestivum formed another subfamily. This result indicated that the members of the two subfamilys have a distinctive difference. In addition, we found the B-hordeins with identical C-terminal end sequences were clustered into a same subgroup (except BXQ053，BZ09-1 and BZ26-5 as a sole group, respectively), so we believe that B-hordein gene subfamilies possibly can be classified on the basis of the conserved C-terminal end sequences of predicted polypeptide and without the limit of SDS-PAGE protein banding patterns. 2. The specific primers were designed according to the published sequences of barley B-hordein genes from Z09 and Z26. Using total DNA isolated from them as the templates, eight clones (designated Z09Pand Z26P) of upstream sequences of the known B-hordein genes was obtained by TAIL-PCR and SON-PCR. Sequences analysis shows that the putative TATA box was present at position –80 bp and CAAT-like box at position –140 bp. Besides, a putative Endosperm Box including an Endosperm Motif (EM) and a GCN4-Like Motif was found at position –300 bp in six clones, and another Endosperm-like box was found at positon –560 bp. While the Endosperm Box or Endosperm-like box was not found in Z09P-2 and Z26P-3. This may indicate that gene expression drived by the two promtors was probably controlled by different trans-acting factors and the genetic control mechanism of corresponding gene expression may be diverse. 3. The B-hordein genic region coding for the mature peptide was cloned into expression vector pET-30a and transformed into bacterial strain BL21 for identifying gene expression fountion. Protein SDS–PAGE analysis showed that only the transformed lysate with the pET-BZ07-2 and pET-BZ26-5 constructs produced proteins related to B-group hordeins of barley, and the mounts of proteins induced by 3 mM IPTG and 3 h were higher than other conditions. This established a base for isolating and putifying B-hordein and further exploring their effects on barley grain quality. 4. The gene-specific primers of B-hordein genes from Z09 and Z26 were used for the quantification of B-hordein gene expression. The α-tubulin gene from Hordeum vulgare subsp. vulgare (GenBank accession number U40042) was used as a control gene. The result shows the transcription of the B-hordein genes in Z09 was found 7 days after flowering, while the transcription of the B-hordein genes in Z26 was found 4 days after flowering, but at a very low level, and it suggested that the B-hordein genes in Z26 probably expressed earlier than those in Z09, or the B-hordein genes in Z09 expressed at so a lower level than Z26 that it can not detected. In addition, B-hordein genes in Z26 accession showed higher expression levels than those in Z09 in four developing stages. Furthermore, a progressive increase in the expression levels of the B-hordein genes between 12 and 18 days after anthesis was observed in both Z09 and Z26. It implies that the B-hordein allelic variants encoded by Hor2 locus exist the differential expression in mRNA levels of during barley endosperm development.

糯小麦杂交后代的基因型鉴定及Wx基因的效应研究

糯小麦在食品加工业、淀粉加工业及其它工业上有着重要用途，是近年来许多国家小麦研究的重要课题。国外糯小麦选育尚未突破高产与糯性相结合的难点，国内目前还没有培育出高蛋白强筋型的糯小麦品种，这在一定程度上与缺乏合适的育种方法和高效、实用的糯小麦分子标记辅助育种技术有关。国内外对Wx基因效应的研究主要利用缺体－四体系、重组自交系或近等基因系，还未见有利用遗传背景相同的BC5F2代回交改良群体的报道。 糯性位点近等基因系是小麦淀粉品质育种的重要材料，而我国目前还没有一套中国栽培小麦遗传背景的糯性位点近等基因系。为了选育部分糯小麦、全糯小麦和中国栽培小麦遗传背景的糯性位点近等基因系，我们利用Wx蛋白电泳和高效实用的分子标记技术体系来鉴定糯小麦杂交后代的基因型，结果证明该体系能有效地用于糯小麦的分子标记辅助育种。以中国春糯性位点全套近等基因系为研究材料，对小麦Wx基因的6个STS标记和1个CAPS标记进行了筛选，改良PCR扩增条件以及产物检测方式后，从这些标记中筛选出3个标记，包括鉴定Wx-A1、Wx-D1位点的2个共显性STS标记和Wx-B1位点的1个显性STS标记。利用上述3个分子标记从BC5F2 代回交改良群体中筛选出了8种Wx基因型，经卡方检验，其分离比符合3对基因的分离比例，其中基因型为aabbdd的植株有2株，直链淀粉含量分别为1.81%和0.82%，为全糯小麦；基因型为AAbbdd，aabbDD的部分糯性植株各有1株，直链淀粉含量分别为15.24%和17.57%。以上4株植株的农艺性状和品质性状接近回交亲本“川育12”，并明显优于全糯材料“98Y1441”，表明采用回交法与Wx基因分子标记辅助选择相结合，有助于培育高产、优质的全糯和部分糯小麦。同时，本研究中建立的分子标记技术体系，也为选育具有中国栽培小麦遗传背景的糯性位点近等基因系奠定了基础。 在基因型鉴定的基础上，利用糯小麦杂交后代BC5F2代回交改良群体研究了各基因缺失降低直链淀粉含量的效果和各基因合成直链淀粉的能力，以及直链淀粉含量与农艺性状、品质性状、淀粉糊化特性等的相关性。缺失不同Wx基因的8种基因型，直链淀粉含量差异显著。研究单缺失基因型发现，减少效应最大的是Wx-B1b基因，Wx-B1b和Wx-D1b基因没有显著差异，减少效应最小的是Wx-A1b基因。研究双缺失基因型发现，Wx基因合成直链淀粉的能力，Wx-B1a基因最高，Wx-A1a基因最低，而Wx-B1a和Wx-D1a基因差异很小；直链淀粉含量与株高、穗长、小穗数、穗粒数、千粒重等农艺性状相关不显著，表明淀粉品质育种可以与高产育种实现有机结合；直链淀粉含量与SDS-沉降值呈显著负相关(r=-0.726)，说明直链淀粉含量降低在一定程度上有利于提高小麦营养与加工品质，这一结果至今未见有文献报道；全糯类型的淀粉糊化特性与其他类型显著不同，具有最高的峰值粘度和稀懈值，最低的低谷粘度、最终粘度、反弹值、峰值时间、糊化温度、起始糊化温度，表明糯小麦淀粉在食品和工业上具有特殊用途；稀懈值与直链淀粉含量呈极显著负相关(r=-0.969)，其他粘度参数与直链淀粉含量呈显著正相关(最终粘度r=0.797，低谷粘度r=0.910、反弹值r=0.954、峰值时间r=0.970、糊化温度r=0.962、起始糊化温度r=0.932)。以上结论可为糯小麦品种选育和淀粉品质改良提供理论依据。 Waxy wheat is very important in food processing industry, starch processing industry and the other industries, so it is a focus of wheat research in many countries these years. Foreign wheat breeders have not conquered the difficulty of high yield combined with waxy character, and there is no waxy wheat variety with high protein and strong gluten in China at present，all of which were caused by lacking proper breeding methods and effective, applied molecular markers-assisted selection technique at a certain extent. Until now, research about the effect of waxy genes mainly depended on nullisomic-tetrasomic lines, recombinant lines or near-isogenic lines, and it is lacking the reports of using improved BC5F2 backcross progenies under the common genetic background. Near-isogenic lines at the Wx loci are important materials for wheat starch quality breeding. However, there are no such lines under the Chinese cultivated wheat genetic background. To develop partial waxy wheats, waxy wheats and near-isogenic lines at the Wx loci under the Chinese cultivated wheat genetic background, we use SDS-PAGE of waxy proteins and effective, applied molecular marker-assisted selection technical system to identify the genotype of waxy wheat’s progenies. The results indicated that such a system is applicable in waxy wheat’s molecular marker-assisted selection effectively. A series of Chinese Spring Wx loci near-isogenic lines were used to identify the specific bands of six STS markers and one CAPS marker of Wx genes. After optimizing PCR amplification and separating of PCR products, three co-dominant and dominant STS-markers were identified at the Wx-A1, Wx-D1 and Wx-B1 loci, respectively, which were used to identify the genotype of waxy wheat’s progenies. Eight Wx genotypes were developed from the improved BC5F2 backcross progenies, which follows Mendelian segregation. Among them, there were two aabbdd waxy plants whose amylose content in the BC5F3 seeds were 1.81% and 0.82%, respectively. In addition, there were partial waxy plants (AAbbdd and aabbDD) whose amylose content in the BC5F3 seeds were 15.24 % and 17.57%, respectively. Most agronomic and quality traits of these four plants resembled those of the recurrent parent “Chuanyu 12”, and superior to waxy wheat parent “98Y1441”. This shows that backcross approach in combination with molecular marker-assisted selection of waxy genes is helpful to develop partial and full waxy wheat with good traits in the waxy wheat breeding program. At the same time, molecular marker-assisted selection technical system in this paper, also establish the base for breeding the near-isogenic lines at the Wx loci under the Chinese cultivated wheat genetic background. According to the results of genotype identification, we use waxy wheat’s improved BC5F2 backcross progenies to verify the effects of null alleles on reducing amylose content and determine the amylose synthesis capacity of each Wx gene independently, and investigate the relativity among amylose content with agonomic traits, quality traits, starch pasting properties respectively. There was significant difference in amylose content of the eight genotypes carrying different null Wx alleles. The reducing effect of the single null alleles was the most significant in Wx-B1b, followed by Wx-D1b and Wx-A1b, and there was no significant difference between Wx-B1b and Wx-D1b. The results of the double null lines further demonstrated that for the capacity of amylose synthesis, Wx-B1a was the highest, followed by Wx-D1a and Wx-A1a, and there was no significant difference between Wx-B1a and Wx-D1a. Amylose contents of the eight genotypes were not significantly correlated with plant height, spike length, spikelets per spike, grains per spike, 1000-grain weight, which showed that starch quality breeding could integrate with high yield breeding. Amylose contents of the eight genotypes were negatively significantly correlated with SDS-sedimentation value(r=-0.726), which suggested that reduction in amylose content is propitious to improve quality at a certain extent. Starch pasting properties of the full waxy type was significantly different from the other seven types, with the highest peak viscosity and breakdown, and lowest valley viscosity, final viscosity, setback, peak time, pasting temperature and starting pasting temperature. It indicated that waxy wheat starch has special use in food and industry. Breakdown was negatively significantly correlated with amylose contents (r=-0.969), and the other parameters were positively significantly correlated with amylose contents (r=0.797 for final viscosity, r=0.910 for trough viscosity, r =0.954 for setback, r =0.970 for peak time, r=0.962 for pasting temperature and r=0.932 for starting pasting temperature ). The results of the study are very useful for waxy wheat variety breeding and starch quality improvement.

A scheme for multiple sequences alignment optimization-an improvement based on family representative mechanics features

As a basic tool of modern biology, sequence alignment can provide us useful information in fold, function, and active site of protein. For many cases, the increased quality of sequence alignment means a better performance. The motivation of present work is to increase ability of the existing scoring scheme/algorithm by considering residue–residue correlations better. Based on a coarse-grained approach, the hydrophobic force between each pair of residues is written out from protein sequence. It results in the construction of an intramolecular hydrophobic force network that describes the whole residue–residue interactions of each protein molecule, and characterizes protein's biological properties in the hydrophobic aspect. A former work has suggested that such network can characterize the top weighted feature regarding hydrophobicity. Moreover, for each homologous protein of a family, the corresponding network shares some common and representative family characters that eventually govern the conservation of biological properties during protein evolution. In present work, we score such family representative characters of a protein by the deviation of its intramolecular hydrophobic force network from that of background. Such score can assist the existing scoring schemes/algorithms, and boost up the ability of multiple sequences alignment, e.g. achieving a prominent increase (50%) in searching the structurally alike residue segments at a low identity level. As the theoretical basis is different, the present scheme can assist most existing algorithms, and improve their efficiency remarkably.

Improvement of the performance of GaN-based LEDs grown on sapphire substrates patterned by wet and ICP etching

Sapphire substrates patterned by a selective chemical wet and an inductively coupled plasma (ICP) etching technique was proposed to improve the performance of GaN-based light-emitting diodes (LEDs). GaN-based LEDs were fabricated on sapphire substrates through metal organic chemical vapor deposition (MOCVD). The LEDs fabricated on the patterned substrates exhibit improved device performance compared with the conventional LED fabricated on planar substrates when growth and device fabricating conditions were the same. The light output powers of the LEDs fabricated on wet-patterned and ICP-patterned substrates were about 37% and 17% higher than that of LEDs on planar substrates at an injection current of 20 mA, respectively. The enhancement is attributable to the combination of the improvement of GaN-based epilayers quality and the improvement of the light extraction efficiency. (c) 2008 Elsevier Ltd. All rights reserved.