葱卵细胞的分离

将大葱(Allium fistulosum)胚珠置于酶液中30分钟可将其外珠被去掉。可清楚地看到由内珠被包裹的胚珠中胚囊的轮廓。将胚珠转移至不含酶的相同溶液中, 用解剖针从胚珠中部切割, 然后挤压胚珠的珠孔部位, 卵器细胞从胚珠的切口处逸出。再用显微操作仪将卵细胞和2个助细胞分开, 达到葱卵细胞分离的目的。酶对分离卵细胞具有重要的作用, 经0.2%果胶酶Y23、0.8%果胶酶、0.8%纤维素酶和0.5%半纤维素酶的处理, 可在2小时内从30 个胚珠中分离出18个卵细胞。随着胚囊的发育, 2个助细胞的体积出现明显差异。生活的葱卵细胞的成功分离, 为建立葱离体受精体系创造了条件。     

洋葱卵细胞的分离

将洋葱的胚珠置于酶液中酶解50-110 min后剥去其珠被,可清楚地看到珠心中的胚囊轮廓。用解剖针将珠心从中部横切,然后挤压其珠孔部位,卵器细胞从胚珠的切口处逸出。再用显微操作仪的玻璃针将卵细胞和两个助细胞分开,达到分离洋葱卵细胞的目的。酶对分离卵细胞具有重要作用,在最佳的酶液浓度[0.02%果胶酶Y23、0.08%果胶酶（Serva）、0.05%纤维素酶和0.05%半纤维素酶]下酶解胚珠110 min后,解剖1 h可从24个胚珠中分离出10个卵细胞（41.67%）。随着胚囊的发育,两个助细胞的体积出现明显的二形性。洋葱生活卵细胞的分离为开展洋葱离体受精建立了基础,也为研究洋葱卵器细胞的发育创造了条件。     

洋葱卵细胞的分离

将洋葱的胚珠置于酶液中酶解50~110 min后剥去其珠被,可清楚地看到珠心中的胚囊轮廓。用解剖针将珠心从中部横切,然后挤压其珠孔部位,卵器细胞从胚珠的切口处逸出。再用显微操作仪的玻璃针将卵细胞和两个助细胞分开,达到分离洋葱卵细胞的目的。酶对分离卵细胞具有重要作用,在最佳的酶液浓度[0.02%果胶酶Y23、0.08%果胶酶(Serva)、0.05%纤维素酶和0.05%半纤维素酶]下酶解胚珠110 min后,解剖1 h可从24个胚珠中分离出10个卵细胞(41.67%)。随着胚囊的发育,两个助细胞的体积出现明显的二形性。洋葱生活卵细胞的分离为开展洋葱离体受精建立了基础,也为研究洋葱卵器细胞的发育创造了条件。     

水稻卵细胞与合子的分离

水稻卵细胞与合子的分离韩红梅赵洁施华中杨弘远周嫦（武汉大学生命科学学院植物生殖生物学研究室武汉４３００７２）关键词水稻,卵细胞,合子,分离ＩＳＯＬＡＴＩＯＮＯＦＥＧＧＣＥＬＬＳＡＮＤＺＹＧＯＴＥＳＩＮＯＲＹＺＡＳＡＴＩＶＡＨＡＮＨｏｎｇＭｅｉＺ...     

分离烟草胚囊的新方法及诱导卵细胞与助细胞原生质体的原位融合

用渗透压冲击和酶解相结合的技术,成功地分离出大量形态完整、结构清晰的烟草生活胚囊。此法用酶浓度低,操作时间短、例于纯化,易于分离胚囊细胞原生质体,为植物雌性细胞操作提供了一种较实用的技术。此外,通过整溶液渗透压,可有效地控制及诱导卵细胞与助细胞原生质体的原位融合。     

蓝猪耳卵细胞和合子的分离

蓝猪耳(Torenia fournieri)胚囊部分裸露出胚珠,在光学显微镜下能清楚观察到卵细胞和助细胞的形态结构.用解剖和酶解-解剖两种方法都能分离出生活卵细胞.用前种方法机械分离出的卵细胞数量较少(5%),但避免了酶对配子识别研究的干扰.在后种方法中加入0.1%纤维素酶和0.1%果胶酶既能使分离更加容易操作,又对卵细胞没有致命伤害,能在短时间内分离出较多的卵细胞(18%).用酶解-解剖方法也可分离出授粉14 h后的合子细胞.     

黄花木本曼陀罗卵细胞分离(简报)

采用分离的精、卵细胞体外融合并诱导人工合子长成植株的离体受精方法可在没有其他组织影响的单细胞水平上探索受精事件的发生过程．为研究高等植物的受精机理、探索配子识别和合子激活等问题提供有效手段。分离的卵细胞不仅可以用来开展离体受精研究．也提供了用分子生物学方法研究被子植物卵细胞发育和合子发育机理的实验基础.     

胡萝卜精、卵细胞、助细胞和中央细胞的分离

用两个解剖针挤压胡萝卜花粉使其破裂释放出精细胞。用酶解-解剖方法分离胡萝卜胚囊中的卵细胞、助细胞和中央细胞。胡萝卜胚珠先在酶液中酶解40~50min,然后将其转移到不含酶的分离液中用解剖针解剖胚珠。将胚珠的合点端切破,轻轻挤压胚珠的珠孔,卵细胞、助细胞和中央细胞即可逸出。在最佳条件下,20min可从20个胚珠中分离出5个卵细胞。对分离胚囊细胞的渗透压和酶液成分进行了筛选。分离出的卵细胞用显微操作仪收集。胡萝卜精、卵细胞的成功分离为在双子叶植物中进行离体受精探索创造了条件。     

浅议仓鼠卵细胞的细胞周期

高中《生物》（必修本）第1册第35页有一个“不同细胞的细胞周期持续时间”表,表中列举了仓鼠卵细胞的细胞周期的具体时间（分裂问期为11．6t／h和分裂期0．8t／h）。不少教师和学生对此提出质疑：仓鼠卵细胞有细胞周期吗？下面就此问题谈谈个人看法。     

莴苣胚囊细胞分离

用酶解和解剖方法分离了莴苣的卵细胞,助细胞,中央细胞和合子。莴苣子房先在酶液中酶解40～50min,然后在不含酶的分离液中用解剖针解剖子房。在解剖出的胚囊中,可看到卵细胞,两个助细胞和中央细胞的轮廓。将胚囊的合点端切破,轻轻挤压胚囊的珠孔端,四个细胞即可逸出。在最佳条件下,90min可从40个子房中分离出29个胚囊,进一步从中分离出11个卵细胞。分离出的胚囊细胞用显微操作仪收集备用。莴苣卵细胞的成功分离为进行离体受精探索创造了条件。     

大葱卵器及受精后助细胞的超微结构

章丘大葱（Ａllium fistulosum L.cv.Zhangqiu)的卵器由１个卵细胞及２个助细胞组成,观察到不少卵器没有卵细胞,只有２个助细胞。卵细胞的核及大部分细胞质位于细胞的合点端,１个大液泡占据了细胞其他部位。卵细胞含有很多的核糖体及多聚核糖体、嵴明显的线粒体、粗面内质网、高尔基体具小泡,卵细胞似是一个活跃的细胞。细胞外被细胞壁,其合点端及侧方与助细胞共同壁不连续,助细胞有一较大的核,位于细胞膨大的部位,众多的小液泡遍布细胞中。核糖体及聚合核糖体、线粒体,粗面内质网及风心圆环状粗面内质丰富,高尔基体及小泡常见,反映了其活跃的代谢作用。助细胞合点端及侧方与卵细胞、中央细胞的共同壁不连续,与卵细胞共同壁含胞间连丝,壁不连续处,有不状多层膜结构伸入卵细胞质,显示助细胞可能对卵细胞提供营养,伟粉后,一个助细胞退化,宿存助细胞至随胚胚期尚存在,它经历了一个缓慢的退化过程,出现质壁分离,细胞质变稀,液泡扩大,细胞器逐渐减少,在椭形胚期,宿存助细胞核内的染色质及核仁消失,有细胞质侵入核内,因宿存助细胞壁变厚,细胞质出现现脂滴,宿存助细胞可能仍有合成功能,宿存助细胞壁出现若干无壁部位,细胞内的营养物质可能通过无壁部位向胚乳转运,供游离核胚乳及胚乳细胞化初期的发育。     

Microtubule Cycle in Root Tip Cells of Allium fistulosum L.

Using immunofluorescent localization techniques and TEM methods, the organization of microtubule arrays during the cell cycle of root tip cells of Allium fistulosum L. was studied. There are four basic types of microtubule organization, namely, interphase cortical microtubule, pre-prophase band microtubule, spindle microtubule and phragmoplast microtubule, which constitute the typical microtubule cycle in dividing cells of higher plants. The fluorescent figures of microtubules observed under fluorescent microscope were explained and analysed by the ultrastractural informations of microtubules obtained from TEM.     

大葱小孢子母细胞至二胞早期花粉发育的超微结构观察

用电镜观察了章丘大葱 (AlliumfistulosumL .)从小孢子母细胞至二胞早期花粉发育的超微结构。终变期的花粉母细胞 ,胼胝壁外方的相邻初生壁间及胞间隙内 ,存在胞间物质 ,四分体期 ,此物质尚部分存在。小孢子母细胞减数分裂前 ,细胞质内含有脂滴 ,小孢子有丝分裂以后 ,脂滴增多增大。小孢子分裂后期 ,质体已积累淀粉粒 1至多个。二胞早期花粉之营养细胞质内 ,有些含淀粉质体亦含脂滴。各发育期 ,核糖体及多聚合糖体丰富 ,并有很多的粗面内质网、高尔基体及小泡、线粒体 ,显示蛋白质、糖类及其它物质合成及运输作用的活跃。小孢子缺中央大液泡。有丝分裂后期 ,细胞器集中于未来的营养细胞极。小孢子胞质分裂期 ,有些内质网贴近或与花粉质膜相连 ,它们或有可能互相融合 ,扩大质膜面积而适应花粉的生长。还讨论了不同时期高尔基体小泡的作用。     

A Comparative Study of Anther and Pollen Development in Male Fertile and Male. Sterile Green Onion (Allium fistulosum L.)

Anther and pollen development in male-fertile and male-sterile green onions was studied. In the male-fertile line, both meiotic microspore mother ceils and tetrads have a callose wall. Mature pollen grains are 2-celled. The elongated generative cell with two bended ends displays a PAS positive cell wall. The tapetum has the character of both secretory and invasive types. From microspore stage onwards, many oil bodies or masses accumulate in the cytoplasm of the tapetal cells. The tapetum degenerates at middle 2-celled pollen stage. In male-sterile line, meiosis in microspore mother cells proceeds normally to form the tetrads. Pollen abortion occurs at microspore with vacuole stage. Two types of pollen abortion were observed. In type I, the protoplasts of the microspores contract and gradually disintegrate. At the same time the cytoplasm of microspores accumulates oil bodies which remain in the empty pollen. The tapetal cells behave normally up to the microspore stage and early stage of microspore abortion, but contain fewer oil bodies or masses than those in the male-fertilt line. At late stage of microspore abortion, three forms of the tapetal ceils can be observed: (1) the tapetal cells with degenerating protoplasts become flattened, (2) the tapetal cells enlarge but protoplasts retractor, (3) the cells break down and tile middle layer enlarges. In type Ⅱ, the cytoplasm degenerates earlier than the nucleus of the microspores and no protoplast is found in the anther locule. There are fibrous thickenings iii the endothecium of both types. It is difficult to verify whether the tapetum behavior and pollen abortion is the cause or the effect.     

RAPDs and noncoding chloroplast DNA reveal a single origin of the cultivated Allium fistulosum from A. altaicum (Alliaceae)

The origin of the crop species Allium fistulosum (bunching onion) and its relation to its wild relative A. altaicum were surveyed with a restriction fragment length polymorphism (RFLP) analysis of five noncoding cpDNA regions and with a random amplified polymorhic DNA (RAPD) analysis of nuclear DNA. Sixteen accessions of A. altaicum, 14 accessions of A. fistulosum, representing the morphological variability of the species, and five additional outgroup species from Allium section Cepa were included in this study. The RFLP analysis detected 14 phylogenetically informative character transformations, whereas RAPD revealed 126 polymorphic fragments. Generalized parsimony, neighbor-joining analysis of genetic distances, and a principal co-ordinate analysis were able to distinguish the two species, but only RAPD data allowed clarification of the interrelationship of the two taxa. The main results of this investigation were: (1) A. fistulosum is of monophyletic origin, and (2) A. fistulosum originated from an A. altaicum progenitor, making A. altaicum a paraphyletic species. Compared with A. altaicum the cultivated accessions of the bunching onion show less genetic variability, a phenomenon that often occurs in crop species due to the severe genetic bottleneck of domestication. Allium altaicum and A. fistulosum easily hybridize when grown together, and most garden-grown material is of recent hybrid origin.     

The Development of Green Onion (Allium Fistulosum L. )Embryo and Endosperm

Embryo development of Zhangqiu green onion conforms to the Asterad type and goes through the following stages: proembryo, globular, ellipsoidal, laterally concave, stick-shaped, and curved and mature. The persistent synergid is present until the late globular stage of embryogenesis. Endosperm development of Zhangqiu green onion follows the nuclear pattern. Endosperm cell formation begins at both the micropylar end and the chalazai end of the embryo sac when the embryo is in the late globular stage. Due to the anticlinal wall formation, a layer of free nuclei becomes a layer of “open cells” which lack the inner periclinat wall. The open cells undergo cell division periclinally, and a layer of complete cells is cut off outside and a new layer of open cells inside. The subsequent cell divisions give rise to the endosperm cells centripetally until those from the opposite of the embryo sac meet. The first anticlinal walls arise from the cell plates without phragmoplasts between the free nuclei in interphase. The first periclinal walls are formed by normal cytokinesis. When a few layers of endosperm cells are formed at the micropylar end and the chalazal end of the embryo sac, free cells are present in the central vacuole.     

Isolation and characterization of lectins and lectin-alliinase complexes from bulbs of garlic (Allium sativum) and ramsons (Allium ursinum)

A procedure developed to separate the homodimeric and heterodimeric mannose-binding lectins from bulbs of garlic (Allium sativum L.) and ramsons (Allium ursinum L.) also enabled the isolation of stable lectin-alliinase complexes. Characterization of the individual lectins indicated that, in spite of their different molecular structure, the homomeric and heteromeric lectins resemble each other reasonably well with respect to their agglutination properties and carbohydrate-binding specificity. However, a detailed analysis of the lectin-alliinase complexes from garlic and ramsons bulbs demonstrated that only the heterodimeric lectins are capable of binding to the glycan chains of the alliinase molecules (EC 4.4.1.4). Moreover, it appears that only a subpopulation of the alliinase molecules is involved in the formation of lectin-alliinase complexes and that the complexed alliinase contains more glycan chains than the free enzyme. Finally, some arguments are given that the lectin-alliinase complexes do not occur in vivo but are formed in vitro after homogenization of the tissue. This revised version was published online in November 2006 with corrections to the Cover Date.