拟南芥bHLH家族转录因子DYT1在花药发育过程中调控胼胝质降解

胼胝质的合成和降解是雄配子体减数分裂过程中的一个重要特征,对后期花粉成熟有重要作用．在此研究中,分离到了一个雄性不育突变体ms157,该突变体的绒毡层分化及胼胝质降解过程出现异常,导致花粉败育．图位克隆和遗传分析表明：MS157基因与bHLH家族转录因子DYT1（At4g21330）是同一基因．因此,将ms157突变体改名为dyt1-2．反式激活作用实验揭示了DYT1的激活功能域位于基因的250～504bp之间．通过酵母双杂交实验发现DYT1蛋白在体内可以形成同源二聚体来执行其功能．RT—PCR及定量PCR分析表明胼胝质酶相关基因A6的表达在突变体背景下严重下调．因此,DYT1通过调控胼胝质的降解来影响花药发育过程．     

转录因子TDF1对拟南芥雄性不育突变体atms1的温敏调控机制研究

TDF1作为MYB家族的转录因子,参与了胼胝质的降解过程,并控制绒毡层的分化从而影响着花粉粒的正常发育.以一株通过甲基磺酸乙酯(EMS)诱变Col-0野生型拟南芥获得的温敏雄性不育突变体atms1(ambient temperature-sensory male sterility1)对温度敏感机制进行了研究.通过构建TDF1反义RNA表达载体,在atms1突变体背景下抑制TDF1的表达来观察突变体花粉育性的变化,发现在27℃时的atms1突变体花粉的育性得以恢复.这一结果表明在花粉发育的过程中TDF1对ATMS1m具有一定的调控作用.     

荧光显微术在鉴别拟南芥花粉壁发育相关基因功能中的应用

花粉在发育过程中,其壁中的不同成分具有自发荧光或诱发荧光的特性,荧光显微术利用此特性来鉴别花粉壁中的化学成分.拟南芥野生型花序的树脂半薄连续切片用苯胺蓝水溶液染色后,在紫外光激发下,胼胝质发出黄绿色荧光,孢粉素发出黄色或黄棕色荧光,纤维素发出蓝色荧光.利用荧光的方法快速简便,分辨率高,适用于大规模筛选花粉壁发育相关基因时对不同基因的功能进行快速鉴别和分类.用该方法鉴别出了几个与胼胝质合成、胼胝质降解、孢粉素沉积模式以及花粉内壁纤维素合成相关的遗传位点.     

拟南芥雄性不育突变体pmil的遗传与定位

通过甲基磺酸乙酯(简称EMS)诱变拟南芥Col -0种子获得一株隐形雄性不育突变体pollen mitosis1,grail.遗传分析结果显示突变体为配子体遗传,细胞学观察发现突变体花粉粒发育出现异常:细胞塌陷,细胞核在单核靠边期后和花粉第一次有丝分裂(简称PMI)前的时间段发生降解,通过图位克隆方法将该基因定位在分子标记T19L18和T1D16之间,物理距离55Kb.测序分析证明这55Kb区间中的ERECTA基因的编码区在3067bp的位置发生单碱基替换,C/G变为A/T.由此说明ERECTA基因在拟南芥从单核小孢子到二细胞花粉的发育过程中起着重要作用.     

拟南芥雄性不育突变体 pmi1 的遗传与定位

通过甲基磺酸乙酯（简称EMS）诱变拟南芥Col－0种子获得一株隐形雄性不育突变体pollen mitosis1,pmil．遗传分析结果显示突变体为配子体遗传．细胞学观察发现突变体花粉粒发育出现异常：细胞塌陷,细胞核在单核靠边期后和花粉第一次有丝分裂（简称PMI）前的时间段发生降解．通过图位克隆方法将该基因定位在分子标记T19L18和T1D16之间,物理距离55Kb．测序分析证明这55Kb区间中的ERECTA基因的编码区在3067bp的位置发生单碱基替换,C／G变为A／T．由此说明ERECTA基因在拟南芥从单核小孢子到二细胞花粉的发育过程中起着重要作用．     

光信号途径组分HEMERA调控拟南芥的花药发育

雄蕊是植物的雄性生殖器官,对植物繁衍及作物的产量至关重要.光信号影响拟南芥生殖发育过程,但作用机制知之甚少.HEMERA(HMR)是拟南芥光敏色素途径的主要成员之一,通过介导光敏色素B(PhyB)的定位及光信号途径重要转录因子PIF的降解而在植物光形态建成过程中发挥重要作用,但其在生殖发育中的功能尚不清楚.本研究利用hmr突变体及转基因恢复植物进行研究发现:HMR功能缺失导致花药变小、花粉数目减少、花粉萌发率显著降低,绒毡层部分空泡化并提前降解;并且,其转基因功能互补能恢复这些育性问题,说明HMR可能通过参与绒毡层降解过程而调节植物雄性育性.     

豌豆PsSGR基因cDNA的分离及其遗传功能分析

拟南芥滞绿基因AtNYE1是衰老过程中调控叶绿素降解的关键基因.利用cDNA末端快速扩增(RACE)技术,在豌豆(Pisum sativum)中分离获得了黄色子叶豌豆PsSGR基因和突变体绿色子叶豌豆PssgrGr基因的全长cDNA.为了验证AtNYE1同源基因PsSGR的功能,将黄色子叶豌豆PsSGR和绿色子叶豌豆PssgrЛ2775的编码区序列分别构建到带有花椰菜花叶病毒35S组成型强启动子和拟南芥叶片衰老特异诱导型启动子SAG12的植物表达载体上,经农杆菌介导、通过花序浸染法转化拟南芥滞绿突变体nye1-1.互补实验结果显示,PsSGR基因能够恢复拟南芥滞绿突变体nye1-1的滞绿表型,而PssgrЛ2775基因则不能.这一结果表明,豌豆PsSGR是负责豌豆子叶中叶绿素降解的调控因子.     

超微结构显示拟南芥花粉壁孢粉素沉积模式及外壁内层的形成

花粉壁由外壁和内壁组成,外壁又分为外壁外层和外壁内层.外壁由绒毡层控制,而内壁由小孢子自身控制.自从有了电子显微镜,就了解到花粉壁有三层结构.花粉外壁内层是高度保守的结构,只有在电镜下才能观察到,但是对其如何形成并不清楚.最近报道了模式植物拟南芥中TEK基因特异调控外壁内层的形成,在该突变体中外壁内层特异缺失而外壁外层能正常形成(Nature communications 5:3855).本文在该项工作的基础上,对模式植物拟南芥花粉壁形成过程进行全面深入分析.发现在四分体的细胞周质中存在的深灰色物质可能是外壁内层成分的前体,其组成成分可能与孢粉素有所不同,当小孢子从四分体中释放时,这些前体物质能够迅速组装成外壁内层.在此基础上,提出了一个拟南芥花粉外壁的发育模型:在减数分裂形成的四分体中,胼胝质壁和小孢子质膜间形成初生外壁.随后小孢子质膜显示出波浪型结构,绒毡层分泌的孢粉素沉积在波浪型质膜顶端发育成外壁外层结构,而绒毡层分泌的外壁内层物质积累在小孢子质膜表面发育成外壁内层结构.四分体胼胝质壁完全降解释放小孢子后,在外壁内层和小孢子质膜间形成内壁.这一包含花粉壁三层结构的模型不仅有助于其他植物物种花粉壁结构和形成过程的了解,也有助于花粉壁分子机理的深入研究.     

拟南芥温敏雄性不育突变体atms1的获得及表型分析

从甲基磺酸乙酯（ethylmethane sulfonate,EMS）化学诱变建立的野生型拟南芥（Col-0）突变体文库中筛选到1株突变体.在低温16 ℃时,该突变体的雄性育性与野生型没有显著差异,花粉染色呈现100%可育.随着培养环境温度的升高,突变体花粉育性逐渐下降,因此,该突变体为一温敏雄性不育突变体,并被命名为atms1（ambient temperature-sensory male sterility 1,即环境温度敏感雄性不育1）.花药切片结果显示,在23 ℃以下,该突变体花药各个发育时期的形态与野生型花药没有显著的差异;而27 ℃处理1周后的突变体花药呈现多种表型：同一朵花中各个花药的发育时期出现显著分化,花粉母细胞胼胝体单薄,绒毡层发育滞后于同时期的野生型花药.遗传分析确定,atms1的不育表型是由单个隐性核基因控制的.     

十字花科植物授粉过程中花粉表达基因的研究

授粉是开花植物有性繁殖的重要过程，这个过程包括物种特异性的花粉与柱头细胞的黏附，干燥花粉的水合，花粉的萌发和花粉管在花柱中的正常生长等，其中涉及花粉众多基因的表达与调控，同时也与柱头细胞中相关基因的活动密切相关．近年来，人们在十字花科植物有性繁殖的分子机制研究中进展迅速，分离鉴定了多个影响授粉过程的突变体及其对应的基因，提出了控制授粉过程各关键步骤的多种可能的机理，在很多方面有了明显的突破．文中以十字花科植物尤其是模式植物拟南芥为例，从分析众多花粉表达基因在授粉过程中的作用入手，介绍了开花植物授粉分子机制研究的最新进展．     

塔中隆起鹰山组岩溶储层特征及主控因素

探讨塔中隆起鹰山组碳酸盐岩岩溶储层发育主控因素，为寻找鹰山组岩性油气藏提供地质理论依据。通过岩芯、薄片观察以及物性资料分析，并结合成像测井解释，认为鹰山组碳酸盐岩为低孔低渗岩溶储层，储集空间主要为溶孔、裂缝及溶洞。将中下奥陶统碳酸盐岩储集层划分为裂缝-孔隙型、裂缝型、溶蚀孔洞型及岩溶洞穴型等类型；精细刻画岩溶储层发育样式，讨论岩溶储层发育主控因素：（1）加里东期中期第I幕致使地层隆升为一个大的宽缓的背斜构造，碳酸盐岩地层普遍遭受顺层淋滤、溶蚀。构造活动的挤压褶皱及断裂作用可形成许多构造裂缝，从而在垂向上增加灰岩地层渗透性和水平方向连通性；（2）泥晶灰岩、含云灰岩、云质灰岩在岩溶过程中，岩性较为致密，只能沿裂缝发生溶蚀作用形成溶孔-扩溶缝，含灰质白云岩和灰质白云岩具有较好孔渗，使得地表水相对更容易流经白云岩地层，形成大量溶孔；（3）白云石化作用普遍提高了白云岩地层（结构好、白云石含量高）的孔渗性能，为岩溶作用提供了基础。     

Effect of internal deletion of Myosin II on growth and development ofDictyostelium discoideum

Four deletion mutantDictyostelium myosin II heavy chain genes, MyΔ824-941 (Δ1/ 3S2), MyΔ934-1454 (ΔS2), MyΔ934-1194 (ΔS2-1) and MyΔ1 157–1454 (ΔS2-2), were transformed by standard electfoporation into mhcA-cells (T-null), a mutantDictyostelium cell devoid of endogenous myosin II heavy chain gene. The growth, development and formation of fruiting bodies of cells expressing those mutant myosin II s under suspension culture were investigated by comparison with the wild type cell. The results indicate that internal deletion of myosin II affeds the growth and development ofDictyastelium. Furthermore, the longer the length of deletion, the more serious the defect in phenotype.     

Plant retinoblastoma homologues control nuclear proliferation in the female gametophyte

Haploid spores of plants divide mitotically to form multicellular gametophytes. The female spore (megaspore) of most flowering plants develops by means of a well-defined programme into the mature megagametophyte consisting of the egg apparatus and a central cell. We investigated the role of the Arabidopsis retinoblastoma protein homologue and its function as a negative regulator of cell proliferation during megagametophyte development. Here we show that three mutant alleles of the gene for the Arabidopsis retinoblastoma-related protein, RBR1 (ref. 4), are gametophytic lethal. In heterozygous plants 50% of the ovules are aborted when the mutant allele is maternally inherited. The mature unfertilized mutant megagametophyte fails to arrest mitosis and undergoes excessive nuclear proliferation in the embryo sac. Supernumerary nuclei are present at the micropylar end of the megagametophyte, which develops into the egg apparatus and central cell. The central cell nucleus, which gives rise to the endosperm after fertilization, initiates autonomous endosperm development reminiscent of fertilization-independent seed (fis) mutants. Thus, RBR1 has a novel and previously unrecognized function in cell cycle control during gametogenesis and in the repression of autonomous endosperm development.     

A sphingosine-1-phosphate receptor regulates cell migration during vertebrate heart development

Coordinated cell migration is essential in many fundamental biological processes including embryonic development, organogenesis, wound healing and the immune response. During organogenesis, groups of cells are directed to specific locations within the embryo. Here we show that the zebrafish miles apart (mil) mutation specifically affects the migration of the heart precursors to the midline. We found that mutant cells transplanted into a wild-type embryo migrate normally and that wild-type cells in a mutant embryo fail to migrate, suggesting that mil may be involved in generating an environment permissive for migration. We isolated mil by positional cloning and show that it encodes a member of the lysosphingolipid G-protein-coupled receptor family. We also show that sphingosine-1-phosphate is a ligand for Mil, and that it activates several downstream signalling events that are not activated by the mutant alleles. These data reveal a new role for lysosphingolipids in regulating cell migration during vertebrate development and provide the first molecular clues into the fusion of the bilateral heart primordia during organogenesis of the heart.     

盘基网柄菌(Dictyostelium discoideum)KAX-3和AK127细胞形态发生和同工酶的比较研究

比较了盘基网柄菌(Dictyostelium discoideum)野生型细胞KAx-3和突变型细胞AK127在多细胞发育过程中的形态发生.两者有明显的区别;突变细胞的发育只能停留在细胞疏松聚集阶段,而野生型细胞能顺利完成整个发育过程.利用SDS-PAGE电泳比较分析了两者在重要发育阶段的三种同工酶:醋酶(EST)、苹果酸脱氢酶(MDH)和谷氨酸脱氢酶(GDH).结果发现AK127细胞发育阶段含有两种醋酶成分a和p,而KAX-3细胞中只有一种醋酶a;而且AK127细胞中苹果酸脱氢酶(MDH)含量在发育后14 h和16 h均高于KAX-3细胞;谷氨酸脱氢酶(GDH)同工酶在发育后14 h时相对含量高于KAX-3细胞,而在发育后16 h含量却低于KAX-3细胞.这些数据显示了两种类型细胞的同工酶有明显的差异;表明突变细胞基因的表达发生了一定的改变,由此说明了gp150分子在盘基网柄菌细胞发育中有重要作用.     

Influence of solution concentration and temperature on the dissolution process and the internal structure of glauberite

Although transport in porous media under the influence of chemistry and temperature is a common phenomenon, the dissolution and internal structure evolution of glauberite during in-situ mining have been unique and challenging. This uniqueness indicates the complexity of mineral dissolutions, whereas the challenge represents the characterization of pore development and evolution during the dissolution processes. To investigate the microstructure development of glauberite under the influence of chemistry and temperature, experimental studies were performed with fine cuboid specimens of 4 mm×4 mm×9 mm soaked in solutions of different concentrations (fresh water, half-saturated, and saturated brine). The evolutions of internal structures were monitored through a micro computed tomography system. The statistical analysis indicated that the concentration and temperature of solutions significantly influenced the evolutions of pore size, porosity, and specific surface area of glauberite. The results showed that the increase in the rates of pore size, porosity, and specific surface area declined with time when glauberite was saturated in fresh water. The main reason for pore parameter variation is the differential concentration of solution. However, in the half-saturated and saturated solutions, the increase in rate increased with time. These observations suggest that the chloride ions contained in the saline solution could facilitate the dissolution of glauberite, whereas the existence of salt effect could contribute to the dissolution of calcium sulfate. Compared with the results at 20℃ and 65℃, the studied parameters of glauberite have dramatically decreased when the mineral was soaked in the solutions at high temperature (95℃). This function was most striking in fresh water. The dissolution of glauberite soaked in fresh water or half-saturated brine solution was conditioned by the temperature and solution concentration. However, the dissolution of glauberite was less influenced by temperature at high concentrations. These findings may feature significant implication for the effective recovery of mineral resources by in-situ solution mining method.     

丙烯基硫脲与银电极表面相互作用的EQCM研究

运用循环伏安(CV)和电化学石英晶体微天平(EQCM)方法研究了酸性介质中银阳极溶出和阴极沉积过程以及丙烯基硫脲(ATU)对该过程的影响.实验结果表明:在酸性硝酸银溶液中,银阳极溶出和阴极沉积过程的实验测定值M w/n分别为18.3和43.4g/mol,尽管实验测定值与银1-电子过程的M w/n理论值(108g/mol)相差甚大,但考虑到酸性介质中氢电极过程的影响,仍可指认:在酸性AgNO3溶液中,银的阳极溶出和阴极沉积过程均为1-电子过程.在含有丙烯基硫脲(ATU)溶液中,Ag阳极溶出和阴极沉积过程的M w/n实验测定值分别为104.1和106.3 g/mol,与Ag的1-电子过程的M w/n理论值非常接近,指示丙烯基硫脲(ATU)并不改变银阳极溶出和阴极沉积过程的机理,但ATU阻化了氢的析出,促进了银的电极过程,加大了电极过程的可逆程度.本文从电极表面质量定量变化的角度提供了Ag阳极溶出和阴极沉积过程的新数据.     

酸性介质中Te电沉积和阳极溶解过程的EQCM研究

运用电化学循环伏安和石英晶体微天平(EQCM)方法研究了酸性介质中Te在Pt电极上电沉积和溶解过程.结果表明:Te的电沉积过程比较复杂,在不同电位区间,其电沉积可以是4-电子、1-电子或者2-电子过程;在较负电位下,Te可进一步还原成H2Te;本体Te阳极溶解过程先为4-电子,后为6-电子过程.本文从电极表面质量定量变化的角度提供了Te电沉积和溶解过程的新数据.     

稠油蒸汽驱过程中储集层伤害模式分析——以辽河盆地西部凹陷齐40区块为例

蒸汽注采过程中的储集层伤害是导致稠油开发后期产量下降的重要因素之一,其伤害机理与伤害模式一直是油藏开发过程中的研究难点。以辽河油田齐40区块莲花油层为例,采用物理模拟实验的方法,分析了蒸汽驱过程中储集层的伤害机理,建立了研究区块蒸汽驱过程中储集层伤害模式。研究结果表明,高温强碱性蒸汽注入后,高温溶蚀区中的矿物溶解量增加,储集层物性明显变好;而中温低溶膨胀区至低温膨胀区中,蒸汽对储集层物性逐渐转为破坏作用,岩石颗粒的溶解量降低,生成的钠基蒙脱石等黏土矿物发生水化膨胀并堵塞孔隙喉道,致使储集层受到伤害。     

Fine mapping and cloning of MT1,a novel allele of D10

Rice tillering is an important determinant for grain production.To investigate the mechanism of tillering,we characterized a multiple tillering mutant (mt1) identified from the japonica variety,Zhonghua 11,treated with EMS.This mutant exhibits advanced tillering development and dwarfed compared with wild-type plants.Genetic analysis and fine gene mapping indicated that the mt1 mutant was controlled by a recessive gene,residing on a 29-kb window on AP003376 of chromosome 1.One putative gene in this region,encoding a carotenoid cleavage dioxygenase 8 (CCD8),was allelic to D10.The mt1 mutant phenotype was complemented by introduction of wild-type MT1,and knockdown of MT1 in wild-type rice mimicked the mutant phenotype.Real-time PCR analysis indicated that the MT1 gene is expressed highly in stems and at a low level in axillary buds,panicles,leaves,and roots.In addition,MT1 expression is clearly under feedback regulation.