The Study of Ultrastructure、ATP、Calcium and RNA-Seq in fertile and sterile anthers of Lycium Barbarum L.

高等植物的雄性不育是杂种优势利用的基础，避免了去雄的繁杂程序。植物雄性不育的种类很多，形成原因也很复杂，对雄性不育机理的探索一直是一个热门研究领域。枸杞雄性不育系是近年来新发现的一种不育类型，具有很大应用潜力，为了明确其雄性不育发生的机制并更好地开发利用这一宝贵种质资源，我们从结构、生理和分子生物学等几个方面展开了研究。 本研究以可育系宁杞1号和不育系宁杞5号为实验材料，用超微结构、ATPase和Ca2+的细胞化学定位，以及转录组和数字基因表达谱研究了枸杞花药发育的特征并比较了不育花药形态、结构和基因表达的差异。主要研究结果如下： (1)超微结构研究结果显示，不育花药最早出现的结构异常是在...The male sterility (MS) of higher plants is the basis for utilization of heterosis. However, there are many types of MS and the reasons caused to it are very complex. The mechanism of male sterility is still unclear, and the research of MS has been a hot research field for a long time. Ningqi 5 is a new type of MS found in Lycium Barbarum L. in 2005. In order to figure out the mechanism of MS a...学位：理学博士院系专业：生命科学学院_发育生物学学号：2162012015381

The Distribution of Calcium in Lycium Barbarum L.during the Development of Anther

摘要 钙是植物生长发育过程中所必需的，它不仅是一种大量营养元素，而且也是一种转导许多生理过程的胞内信号分子，有很多重要的生理功能。高等植物的有性生殖是一个非常复杂的发育过程，包括一系列连续的发育事态。钙参与植物有性生殖过程中的许多环节，在被子植物有性生殖过程中钙的功能也越来越多地被揭示出，但是早期的实验主要集中在：1.离体条件下钙对花粉萌发和花粉管生长的影响；2.花粉管顶端梯度分布的游离钙与花粉管生长的关系；3.开花前后胚珠中的钙与花粉管定向生长的关系。在雄性生殖系统中，花药的发育和钙的分布特征存在什么关系？这方面的报道还比较缺乏。本实验采用焦锑酸盐沉淀法，以宁夏枸杞（Lyciumbarba...Abstract Calcium is an indispensable element during plant growth and development, which not only functions as a signal molecule but also a nutritious element with many important physiological functions. Sexual reproduction of higher plants is a very complex process including a series of delicate differentiations of sexual organs and cells. Ca2+ is of importance in the process of sexual reproducti...学位：理学硕士院系专业：生命科学学院生物学系_发育生物学学号：20042605

The Character of Anther Development of Impatiens balsamina

凤仙花花药发育比较特殊:在造孢细胞时期,花药横切面中央是体积较大、细胞内含物较多的细胞团、包括造孢细胞和绒毡层细胞。花药药壁细胞的细胞质较稀少,与中部细胞界限明晰。花粉母细胞时期的花药药壁由约6层细胞组成,但细胞的界限不明显;绒毡层细胞显示变形流入药室中。到四分体时期,绒毡层细胞进一步退化。开花时,成熟花药的药壁细胞由一层表皮细胞、两层药室内壁细胞和一层中层细胞组成。对凤仙花花药绒毡层的特殊性质进行了讨论。The anther development of the Impatiens balsmina displayed some special characters.In sporogenous cell stage, there are some large sporogenous cells, containing density cytoplasm, locating in the central of anthers.The cells of anther wall contain less cytoplasm which makes both distinguish.In the stage of microspore mother cell, the anther wall consists of 6 layers of cells, but the limit line is not clear between the cells.The tapetal cells begin to display the degeneration at this time.In tetrad stage, the tapetal cells degenerate further as the cells become small.The mature anther wall consists of four layers of cells: one layer of epidermis, two layers of endothecium, and one layer of middle layer.The special characters of tapetal cells of Impatiens balsamina were discussed.国家自然科学基金(31170289;31100225); 公益性行业(农业)科研专项课题(200903016)资

Calcium Distribution in the Stigma and Germinating Pollen of Lycium barbarum

用焦锑酸钾沉淀法对宁夏枸杞柱头和花粉中的钙离子分布进行了研究.结果显示,宁夏枸杞柱头表皮有一覆盖层,其中有许多含钙沉淀颗粒的小泡,当花粉落到柱头后从覆盖层中吸水,在萌发孔的表面上聚集了较多的钙沉淀颗粒.同时,花粉内部出现许多含钙的小液泡,使花粉体积增大,内部产生膨压,花粉萌发;生长在覆盖层中的花粉管顶端穿过覆盖层小泡时,附近聚集了较多的钙沉淀颗粒,在花粉管壁上也附着较多的细小钙沉淀颗粒.萌发的花粉粒中由大液泡占据,在其亚顶端的细胞质中,聚集较多钙沉淀颗粒的线粒体膨大形成了一些含钙沉淀颗粒的小液泡,由这些小液泡融合形成的大液泡,将花粉管细胞质挤到其顶端,使其极性生长.这是首次发现在植物柱头覆盖层中有钙离子的现象,从体内证明了钙离子在花粉萌发过程中的现象.讨论了枸杞柱头组织中钙的分布和花粉管的萌发与生长的关系.The stigma of Lycium barbarum is wet type and some material to be secreted to the surface of stigma to form a layer of tectum at anthesis.There were many vesicles in the tectum and numerous calcium precipitated grana were accumulated in the tectum vesicles.When pollen grains landed onto the stigma they began to hydrate and form many small vacuoles in which some calcium precipitated grana located,which made a swelling pressure in pollen.During pollen germinating in the tectum,the pollen tubes often traversed tectum vesicles and displayed many calcium precipitated grana on the tube wall,suggesting it to absorb calcium of the tectum vesicles.In the base of pollen tube some mitochondria with calcium precipitated grana swelled to form small vacuoles which fused to form a large vacuole.The pollen tube cytoplasm was pushed into the top of tube to make sure directional growth.This is first time to report that stigma tectum containing rich calcium in flowering plants.The calcium distribution in the stigma and pollen germination and tube growth were also discussed.国家自然科学基金项目(30670126

Big⁃data based research on correlation between mental flexibility and healthy personality of undergraduates

目的运用MMPI-399量表中的不同因子分和中国常模资料作为健康人格的相关指标来考察它们与AAQ-Ⅱ量表总分和CFQ-F量表总分的相关性,将MMPI-399与SCL-90量表总分的相关性程度的作为比较指标,探讨心理灵活性与MMPI-399各因子的相关程度及其对健康人格的预测作用。方法采用心理灵活性模型中的认知融合问卷(CFQ-F)、接纳与行动问卷第2版(AAQ-Ⅱ);以及MMPI-399人格量表、SCL-90量表、自编人口学调查表,对黄山学院全体在校大学生进行心理健康普测,按照MMPI量表中疑问量表原始分小于22分,谎言量表原始分小于10分的标准,收回有效问卷8 235份。结果大数据视域下,心理灵活性在不同年级和性别的人中有差异;相较于SCL-90总分,心理灵活性相关量表与MMPI-399各因子均有较高的相关性(P&lt;0.01);回归分析结果显示,心理灵活性对抑郁人格均有显著地预测作用(P&lt;0.01)。结论个体心理灵活性存在差异,与健康人格形成相关性显著,心理灵活性对健康人格有很好的预测作用。</p

Detection of distribution change and diffusion of suspended sediment concentration around Heini Bay

利用2004年冬季对黑泥湾近岸海域悬浮体调查资料及2个时相的遥感影像,探讨了该海域悬浮体的分布和扩散特征,分析了影响悬浮体分布的主要因素和悬浮体的主要物质来源.结果表明,影响本海域悬沙质量浓度及其分布的主要因素是风浪掀沙和潮流.夏季,近岸悬沙质量浓度高,向海逐渐减少;冬季,悬沙质量浓度近岸低,向海逐渐增大.由于本海域没有大的河流泥沙汇入,悬沙的主要物质来源是沿岸侵蚀、冲沟排沙和海底泥沙再悬浮

Histochemical Study of Polysaccharides and Lipids on the Developing Anthers of Impatiens balsamina

以不同发育时期的凤仙花花药为实验材料,采用组织化学方法,对花药发育中的结构变化及多糖和脂滴物质分布进行观察。结果表明:(1)凤仙花的花药壁由6层细胞组成,包括1层表皮细胞,2层药室内壁细胞,2层中层细胞和1层绒毡层细胞。其中绒毡层细胞的形态不明显,很难与造孢细胞区分,且在小孢子母细胞时期退化。(2)在小孢子母细胞中出现了一些淀粉粒,但减数分裂后,早期小孢子中的淀粉粒消失,又出现了一些小的脂滴;随着花粉的发育,小孢子形成大液泡,晚期小孢子中的脂滴也消失;小孢子分裂形成二胞花粉后,营养细胞中的大液泡降解、消失,二胞花粉中又开始积累淀粉;接近开花时,成熟花粉中充满细胞质,其中包含了较多的淀粉粒和脂滴。(3)在凤仙花的花药发育中,绒毡层细胞很早退化,为小孢子母细胞和四分体小孢子提供了营养物质;其后的中层细胞退化则为后期花粉发育提供了营养物质。The anther development of Impatiens balsamina displays some special characters:(1)Anther wall consists of 6 layers of cells:one epidermis,two endotheciums,two middle layers and one tapedum.Tapetal cells are not clear during sporogenous stage and difficult to distinguish with sporogenous cells.The tapetal cells degenerate during the stage of microspore mother cell(MMC).(2)Some starch polysaccharides appear in the microspore mother cells which had formed a callose wall.After meiosis of MMCs,starches in early microspores disappear,and lipids were accumulated in early microspores.With microspore developing,late microspores formed several large vacuoles and lipids disappeared.After microspore division,comparing with the big vacuoles in vegetative cell disappearing,starches again appeared in the 2-cellular pollen.Nearly anthesis,mature pollen grains of I.balsaminaaccumulated many lipids and starches as storey materials.(3) During anther development of I.balsamina,some cells of anther wall degenerated and their cytoplasm transformed lipids for pollen absorbing,the tapetal cells degenerated and offered lipids for MMCs and microspores,and middle layer cells for bicellular pollen grains.国家自然科学基金(31100225); 公益性行业(农业)科研专项课题(200903016); 浙江省自然科学基金(Y3110395

ATPase DISTRIBUTION IN FERTILE AND STERILE ANTHER OF A GENIC MALE STERILE CHINESE CABBAGE

对白菜核雄性不育两用系可育花药和不育花药的ATPase做了定位分析。可育花药的花粉母细胞核中积累了大量的ATPase反应颗粒,而细胞质中ATPase反应颗粒较少,但在线粒体中特异地聚集了一些大的ATPase反应颗粒。减数分裂后,小孢子细胞质中ATPase反应颗粒明显增加。随着小孢子发育,其细胞质中ATPase反应颗粒逐渐减少,但在线粒体中又特异性地聚集了较多的AT-Pase反应颗粒。当花药发育到二胞花粉时期,花粉和绒毡层细胞中的ATPase反应颗粒已很少了。不育花药的花粉母细胞中呈现较多的ATPase,然而在线粒体中很少。异常四分体小孢子细胞质中虽然有较多的ATPase反应颗粒,但还是通过细胞质收缩和质壁分离方式退化。对可育花药的花粉母细胞线粒体中特异出现的簇状ATPase分布现象进行了分析,讨论了不育花药中花粉母细胞线粒体ATPase的异常与花粉败育的可能关系。Lead precipitation technique was used to locate Adenosine Triphosphatase (ATPase)in the fertile and sterile anthers of a genic male sterile Chinese cabbage (Brassica campestris L. ssp. Chinensis Makino var. communis Tsen et Lee),which would help us to understand the relationship between ATPase and sterility of anthers of the cabbage. At megaspore mother cell (MMC) of fertile anther many ATPase reactive precipitates were located in nucleus but few of the precipitates in cytoplasm of the cell. Meantime, some ATPase reactive precipitates also specially appeared in mitochondria of the MMC. After meiosis of MMC, the precipitates in cytoplasm of early microspores increased evidently and then decreased step by step with development. The ATPase reactive precipitates in tapetal cell also increased ultimately in early microspore stage and then decreased with development of anther. When microspore formed a large vacuole, which is late stage of microspore, the ATPase reactive precipitates were located in its mitochondria. After microspores mitosis a few of the ATPase reactive precipitates appeared in pollen grains and tapetal cells. More ATPase reactive precipitates appear in MMC of sterile anther than in fertile anther but fewer of them in mitochondria. Although more ATPase granules appear in abnormal tetrad microspores which degenerate by cytoplasm shrinkage and plasmolysis. The relation between ATPase and male sterility of the cabbage was discussed.国家自然科学基金(30570104

CALCIUM DISTRIBUTION IN THE DEVELOPING ANTHER OF LYCIUM BARBARURN L.

在枸杞花药发育过程中,用焦锑酸钾沉淀的钙颗粒显示出了一个与花药发育事件有关的分布特征：在孢原细胞时期的花药中钙颗粒很少。在造孢细胞到小孢子母细胞时期,花药中钙颗粒增加。当花粉母细胞进行减数分裂时,花药中的钙颗粒进一步增加,尤其是在小孢子母细胞的胼胝质壁中。在小孢子发育早期,花药药隔部位的绒毡层细胞质中钙颗粒也明显增加并特异性地分布在其内切向壁上。当小孢子被释放出后,钙颗粒开始特异性积累在正在形成的花粉外壁中,尤其在萌发孔的部位聚集了大量的钙颗粒。当小孢子形成大液泡时,其细胞质中的钙颗粒明显减少。在小孢子分裂形成二胞花粉后,在二胞花粉的大液泡中又特异性地出现许多细小钙颗粒。随着二胞花粉的大液泡完全消失,其细胞质中又出现了许多钙颗粒。接近开花时的成熟花粉粒细胞质中,细小的钙颗粒主要分布在营养细胞和生殖细胞中。枸杞花药发育过程中钙的分布特征反映了其参与调控花粉发育过程。We used potassium antimonate to precipitate“exchangeable cellular Ca~(2+)”-calcium that is sufficiently loosely bound to combine with antimonite,to investigate the feature of calcium distribution during anther development of Lycium barbarurn L.Before the stage of microspore mother cell,few calcium-induced precipitates were found in sporogenous cells and the somatic cells of anther wall.When microspore mother cell (MMC) preparing meiosis,calcium precipitates appeared in the cytoplasm of tapetal cells and callus wall surrounding MMC.After the meiosis of MMC,abundant calcium precipitates were accumulated in the cytoplasm of early microspores,and then in pollen wall,especially in the part of germ-pores.During the late microspore stage,a big vacuole formed and the nucleus was forced to move to peripheral region.Calcium precipitates de- creased sharply and might dissolve in the large vacuole.After microspore mitosis,calcium precipi- tates appeared in the big vacuole of 2-cellular pollen,and then the vacuole disappeared.After that,the calcium precipitates again appeared in the cytoplasm of 2-cellular pollen,and the cyto- plasm became densely and storage materials like starches accumulated inside the pollen grains. When pollen maturating,many small calcium precipitates distributed in its cytoplasm,especially in nucleus.The feature of calcium distribution in the anther of Lycium barbarurn L.means that it plays some biological roles during microspore development.国家自然科学基金(30670126)