银杏精子细胞生毛体及其它细胞器的超微结构

生毛体与嗜锇颗粒是银杏 (Ginkgobiloba)精子细胞中最具有标志性的结构。生毛体是细胞质内一直径为 3～ 4μm的圆球形结构 ,它由一个电子致密的核心和由此向周边发散出的辐射状中心粒组成 ,致密核心上具有微管结构的弱电子染色区域 ,并有微管从生毛体延伸到细胞质。嗜锇颗粒直径为 1 0～ 2 0μm,呈圆球状 ,位于生毛体和细胞核之间 ,其相对的另一侧存在纤维颗粒体。在精子细胞质内 ,特别是嗜锇颗粒和生毛体周围线粒体、质体、内质网、高尔基体等细胞器丰富。银杏精子细胞核较大 ,在细胞核内 ,核仁结构呈球形 ,电子染色致密的颗粒区在周围 ,而纤维组分则在圆球的中间。在核膜表面布满了分布不均匀的核孔复合体。     

银杏精细胞结构及受精作用的细胞学研究

采用常规石蜡制片技术和环氧树脂半薄切片技术,对银杏的精细胞结构及受精过程进行了研究。结果表明,临近受精前,精原细胞分裂形成两个半球形的精细胞,每个精细胞内含液泡状结构、生毛体和纤维性颗粒体各一个。两精细胞的液泡状结构其位置可同时或分别位于近极面和远极面,这种位置的变动可能是鞭毛摆动导致精细胞的旋转所引起,这说明银杏的精细胞在花粉管内也许即可以旋转运动。在银杏受精前后,珠孔端颈卵器室附近的珠心组织表现出向上隆起、出现受精滴、皱褶等规律性变化,这些现象是判断银杏受精时期的良好形态特征。在银杏受精时带有鞭毛的完整精细胞进入颈卵器,随后鞭毛带及精细胞质遗留于颈卵器口下方、卵细胞上面,仅精核进入卵细胞。进入卵细胞的精核直径约30μm,小于成熟精细胞的精核直径（约40μm）。这些对探讨银杏的系统地位及裸子植物的生殖演化具有一定意义。     

一株银杏内生真菌菌株的抑菌活性成分研究

从银杏叶柄分离筛选到具抗菌活性的内生真菌Colletotrichum.SP NTB-2菌株,利用硅胶柱色谱、制备高效液相色谱等方法在其发酵产物中分离到抗枯草芽孢杆菌、鼠伤寒沙门氏菌等具有广谱抑菌活性的化合物,经MS、NMR等波谱数据确认该活性成分为芹菜素-8-C-葡萄糖苷(apigenin-8-C-β-D-glucopyranoside),该化合物首次从真菌中分离得到。     

银杏叶片形态研究

通过对银杏不同枝类、不同叶序的叶片形态进行调查,结果表明:银杏不同枝类的叶片形态差异较大,叶宽、叶柄长、叶基角:多年生鳞枝＞一年生鳞枝＞一年生长枝;叶长、叶形指数:一年生长枝＞一年生鳞枝＞多年生鳞枝;叶面积:一年生和多年生鳞枝＞一年生长枝;有缺刻叶比例:一年生长枝＞多年生鳞枝＞一年生鳞枝.一年生长枝叶片的叶宽、叶长、叶面积、叶柄长、叶基角均随叶序的增加逐渐减小,叶形指数和有缺刻叶的比例则增加.一年生和多年生鳞枝1～5叶的叶长、叶宽和叶面积随叶序增加而逐渐增加,第5～6叶达最大,以后随叶序增加而逐渐减小,叶形指数和叶柄长度随叶序增加而增加,叶基角随叶序增加而减小.一年生长枝的第2叶、一年和多年生鳞枝的第4叶可作为品种描述的标准叶.     

银杏内生真菌的生态分布

2004—2005年春夏秋3季,在江西省九江地区4个地点分别采集健康的银杏(Ginkgo biloba)植株,分离得到内生真菌198株,经形态学分类鉴定为6目、7科、22属。研究结果表明,银杏内生真菌的分布存在季节差异、组织部位差异、年龄差异;优势菌群的组成与数量随采样地点、分离组织部位、不同生长时期内生真菌的不同而变化。与中国其他地区的银杏植株内生真菌进行比较,结果表明,植物内生真菌的分布与地域具有一定的相关性,体现了内生真菌生态分布的多样性。     

Ginkgo biloba: The Ovuliferous Leaf and Its Phylogenetic Implication

This paper reports the occurrence of the ovuliferous leaves on a Maidenhair tree (Ginkgo biloba)growing in Longdong, Yiyuan County, Shandong Province. This phenomenon is discussed in connection with the history of Ginkgoales. It is believed that this discovery serves as an illustration of the hypothesis that the secondary apical position of ovules (Ginkgoales) is a modification of the marginal position (Pteridospermidae). Again, it probably leads to the assumption that, in Ginkgo, “the foliage leaves” bear some features of “thesporophylls” and they are derived from telomes.     

银杏分子技术的研究进展

综述了近些年来分子技术在银杏研究中的应用情况,对银杏的遗传多样性、进化地位、品种鉴别、遗传连锁图谱、雌雄鉴别及其它相关研究进展进行了介绍,并对当前研究中存在的问题和今后的发展方向作了进一步探讨。     

银杏性别相关分子标记

利用RAPD技术寻找银杏(Ginkgo biloba L.)中与性别相关的分子标记。筛选了1200个10bp的随机引物,产生了8372个RAPD条带。只有S1478产生一条大小为682bp、雄性特异的分子标记,该分子标记被命名为S1478—682,出现在所有雄性植株中,而所有雌性植株都不具有该分子标记。通过在北京和沈阳种植的银杏植株的RAPD推广验证,说明该分子标记可以用来检测银杏植株的性别。     

HPLC法测定银杏叶征剂中决黄酮甙的含量

银杏 (ＧｉｎｋｇｏｂｉｌｏｂａＬｉｎｎ .)叶提取物 (ＧＢＥ)是银杏叶经提取、分离而得的干浸膏 ,是银杏叶制剂的主要原料。药理及临床表明 ,它能增加冠状动脉血流量 ,可用于动脉硬化及高血压所致的冠状动脉供血不全、心绞痛、心肌梗塞、脑血栓、脑血管痉挛等症[1] 。其活性成分主要是黄酮类和萜内酯[2 ] 。银杏叶黄酮类成分的测定方法 ,有紫外分光光度法和高效液相色谱法 ,因前者专属性较差 ,现多数厂家均采用后者。高效液相色谱法测定总黄酮甙的含量时 ,在对照品的使用上 ,既有单用槲皮素 (ｑｕｅｒｃｅｔｉｎ)为对照品的测定方法[3 ] ,…     

银杏药用物质的开发利用现状及其发展设想

本文概述了银杏叶的药理作用及其开发利用现状。目前,银杏药用物质的获取仍然是通过叶片提取这一途径,这同时存在地理环境、季节等诸多限制因素。而通过银杏组织、细胞培养获得其药用物质的技术研究刚刚起步,距离实际应用仍有相当的距离,但随着这项技术的不断发展,它不仅可用于生产银杏黄酮类和银杏内酯,而且有助于进一步研究银杏所含的特殊化学成分及其相关的代谢途径。     

银杏性别相关分子标记

利用RAPD技术寻找银杏(Ginkgo biloba L.)中与性别相关的分子标记.筛选了1 200个10 bp的随机引物,产生了8 372个RAPD条带.只有S1478产生一条大小为682 bp、雄性特异的分子标记,该分子标记被命名为S1478-682,出现在所有雄性植株中,而所有雌性植株都不具有该分子标记.通过在北京和沈阳种植的银杏植株的RAPD推广验证,说明该分子标记可以用来检测银杏植株的性别.     

Antiestrogenic activities of Ginkgo biloba extracts

Most climacteric and postmenopausal women appear to have vasomotor symptoms as well as a high risk of osteoporosis and cardiovascular disease. Although exogenous estrogens can reduce these symptoms, women are reluctant to use hormone replacement therapy (HRT) due to its undesirable side effects, such as irregular bleeding and an increased risk of breast cancer. A previous study suggested that Ginkgo biloba extracts (GBE) have estrogenic activity and might be suitable as an alternative to HRT. However, there are no reports of the preventive effect of GBE on breast cancer, which is the side effect of classical HRT. In this study, it was confirmed that GBE exhibits estrogenic and antiestrogenic activity depending on the E₂ and GBE concentration, via estrogen receptor (ER)-dependent and ER-independent pathways. In addition, GBE reduced the E₂ levels by stimulating the E₂ metabolism and inhibiting E₂ synthesis, which indicates that GBE can induce antiestrogenic activity via the depletion of E₂. Furthermore, GBE might have similar action to selective arylhydrocarbon receptor modulators (SAhRMs), which induce antiestrogenic activity through cross-talk between the arylhydrocarbon receptor (AhR) and ER. In conclusion, GBE has a biphasic effect on estrogen, and can be considered as a potential alternative to HRT with chemopreventive effects on breast cancer. However, further studies on animals and humans will be required.     

银杏(Ginkgo biloba)叶表皮特征及其气孔的发育

通过光学显微镜和扫描电子显微镜,对离析和未离析的、成熟的和刚开始发育的银杏叶表皮进行了研究.表皮基本特征:两面生气孔型,但气孔主要分布在下表皮,上表皮气孔的数目和分布因个体而异;气孔器为单唇单环式,围绕每个气孔的副卫细胞数目多为5～6个;副卫细胞的平周壁明显加厚,具乳突状突起,拱盖着下陷的保卫细胞;偶有双气孔发生;上表皮厚于下表皮;表皮细胞可明显分为叶脉区和脉间区.角质外蜡质层:在高倍扫描电子显微镜下,管状的蜡质晶体交错覆盖在叶片的外表面,还观察到有些管状的蜡质晶体呈分叉状态.气孔的发育:通过我们的观察,认为银杏气孔的发生属于典型的周源型, 即副卫细胞与保卫细胞各自发生于不同的母细胞,而不是前人报道的中--周型起源.     

Cytoskeleton in Pollen and Pollen Tubes of Ginkgo biloba L.

The distribution of F-actin and microtubules was investigated in pollen and pollen tubes of Ginkgo biloba L. using a confocal laser scanning microscope after fluorescence and immunofluorescence labeling. A dense F-actin network was found in hydrated Ginkgo pollen. When Ginkgo pollen was germinating,F-actin mesh was found under the plasma membrane from which the pollen tube would emerge. After pollen germination, F-actin bundles were distributed axially in long pollen tubes of G. biloba. Thick F-actin bundles and network were found in the tip of the Ginkgo pollen tube, which is opposite to the results reported for the pollen tubes of some angiosperms and conifers. In addition, a few circular F-actin bundles were found in Ginkgo pollen tubes. Using immunofluorescence labeling, a dense microtubule network was found in hydrated Ginkgo pollen under confocal microscope. In the Ginkgo pollen tube, the microtubules were distributed along the longitudinal axis and extended to the tip. These results suggest that the cytoskeleton may have an essential role in the germination of Ginkgo pollen and tube growth.     

Callus Formation at Graft Interface in Ginkgo biloba L.

The early stages of graft union, when male branch was grafted onto female branch in Ginkgo biloba L. by cleft graft, have been observed under light microscope in order to determine the origin of callus cells between the stock and scion. Pith parenchyma cells near the graft interface were the earliest cells in response to such method of grafting. These cells dedifferentiated and then divided within 7～ 12 days after grafting. A large number of callus cells extended from the pith into the space between the graft interface linking the stock with the scion about 18～ 20 days after grafting; and then continued to proliferate and extend outwards along the space. Cambium cells and immature vascular tissue near the graft interface dedifferentiated into callus rather late. Theover all link between the stock and scion was completed in the sites 30 days after grafting. Callus cells were also produced from corticai parenchyma cells, but they were much limited in quantity. In conclusion, the graft interface may be considered as a "natural culture bed" after grafting, in which all undamaged, living cells are capable of dedifferentiation and producing callus cells for compatible graft union. In the case of G. biloba (male/female) it were the pith parenchyma cells that appeared first to form the callus cells and later extend to link the stock with the scion.     

Development of microsatellite markers in ginkgo (Ginkgo biloba L.)

Ginkgo biloba L. is one of the oldest unevolved tree species on Earth. We isolated five polymorphic microsatellite loci from G. biloba using a dual‐suppression polymerase chain reaction technique. These loci provided microsatellite markers with high polymorphism ranging from three to 13 alleles per locus. The observed and expected heterozygosities ranged from 0.667 to 0.952 and from 0.640 to 0.897, respectively. The markers will contribute to research on the conservation, genetic diversity and mating patterns of G. biloba.     

银杏套细胞发育的解剖学研究

银杏套细胞起源于近雄配子体表达的颈卵器母细胞周围细胞,中央细胞形成时,套细胞呈明显的一层,围绕颈卵器紧密排列,随着颈卵器的发育,套细胞体积逐渐地大,细胞质变浓厚,细胞质中脂滴增多,套细胞和中央细胞的接触壁开始出现局部加厚,在颈卵器发育的泡沫化阶段,套细胞和中央细胞的接触壁不均匀加厚较为显著,在较薄的区域可见胞间连丝,受精前,套细胞中液泡增多,脂滴迅速减少,造淀粉体增大,套细胞与卵细胞的接触壁的不均匀加厚非常明显,精核进入颈卵器以后,卵细胞与套细胞的接触壁和卵细胞的质膜之间形成一个薄厚不均的间隔层,受精卵分裂时,受精卵细胞与套细胞接触壁的凹陷处可见许多小泡和内质网,游离核期时,套细胞内出现大量小液泡,细胞内含物迅速消失,套细胞外形变长,胚胎长出颈卵器后,套细胞逐渐解体消失。     

银杏种子和叶的蛋白质分析

采用分级提最方法提取银杏（Ginkgo biloba L.）种子和银杏叶蛋白质,并进行各组分蛋白质含量测定和银杏种子后熟过程中蛋白质含量动态变化的分析。结果表明,银杏种子以水溶性和盐溶性蛋白质为主,银杏叶以醇溶性蛋白质为主。银杏叶中主要蛋白质在HPLC柱中的保留时间为3.457min,相对含量达70%以上。银杏叶蛋白质含有丰富的亮氨酸、缬氨酸、赖氨酸和色氨酸,其含量分别为20.23%、13.35%、4.81%和3.73%。萌发种子胚体中的蛋白质主要是醇溶性蛋白质和谷蛋白类蛋白质。在种子萌发过程中,胚乳蛋白质含量明显增加,播种后第3周和萌发时总蛋白质含量达到高峰。     

银杏雌株染色体Ag-NOR的研究

本文以银杏雌株的幼小胚珠为材料,对其体细胞染色体进行了Ag-NOR染色研究。发现其Ag-NOR(随体)的数目均为4个,大小则有同形和异形两种类型。认为NOR(随体)的数目和形态不能作为性染色体的识别特征,并对NOR的杂合性和多态性进行了讨论。 Abstract:In this paper,the Ag-NOR of somatic chromosomes of young ovulum in Ginkgo biloba was studied.It was found that the number of Ag-NORs(satellite)were four constantly,though there were two kinds in size,homoeotype and heterotype.So we considered that the number and morphology of NOR(satellite)can not be used as identification markers of sex chromosome in Ginkgo biloba.In addition,the heterozygosity and polymorphism of NOR are discussed.     

An embryological study and systematic significance of the primitive gymnosperm Ginkgo biloba

Ginkgo biloba L.is considered one of the most ancient seed plants,with several primitive features of plant reproductive process.However,the phylogenetic position of Ginkgo and its relationship with other extant seed plants remain unclear.To gain a better understanding of these issues,we observed the embryological development of G.biloba using semi-thin sections and scanning electron microscopy.In late August,the zygote moved from the end of the micropylar to the middle of the archegonium,and mitosis resulted in many free nuclei distributed randomly in the archegonium.Afterwards,the cell wall was formed and the proembryo began to differentiate into the embryonal region and the underdeveloped presuspensor region.In early October,the embryo differentiated into two cotyledons,plumule,hypocotyl,radicle,and suspensor tissues.Subsequently,the two cotyledons grew rapidly,but the undeveloped suspensor began to degenerate and gradually disappear,indicating that the embryo had begun to mature.During early embryo development,the main supply of nutrients was carbohydrate in the cells of the jacket,tentpole,and surrounding endosperm,whereas endosperm provided nutrients during embryo maturation.Our results indicate that Ginkgo is extremely similar to cycads in terms of embryology but more similar to conifers in macromorphology and vegetative anatomy,suggesting that the Ginkgo lineage may have an intermediate phylogenetic position between cycads and conifers.