玉米胚胎发育、萌发与胚的结构及子叶二型性

运用扫描电镜与半薄切片技术,观察了玉米(Zea mays L.)的胚发育过程,得到以下认识：第一、关于原胚。玉米合子细胞分裂形成的原胚分为胚柄、胚基与胚体三部分。胚柄短小,寿命短暂。胚基具有生长带,纵向伸长长度大,胚基的上部参与形成胚根鞘,其余部分干缩后附在胚根鞘末端。第二、玉米胚的背腹极性及二型子叶。原胚初期胚体出现背腹极性,腹面的细胞小,细胞质稠密,液泡较少;背面的细胞较大,细胞质稠密度略低,液泡较多。原胚后期胚体分化为腹部与背部,腹部从腹面的中央突起,背部在腹部的周围(从左至右侧)及整个胚体背面。进入幼胚时期,腹部分化为胚芽鞘、生长锥、胚轴、胚根和胚根鞘(大部分)。期间,胚芽鞘原基和根原始细胞的分化都从胚体的中轴部位开始,然后向两侧和四周扩展,表现出胚体腹面形态的两侧对称性。原胚的背部形成盾片原基,盾片原基经历向左、右、上、下的迅速扩展和加厚的生长,将整个腹部所分化形成的构件藏于盾片的纵沟之中,最后盾片从纵沟的边缘长出的左、右侧鳞均向胚体的中轴线生长,完整显示出玉米胚腹面的两侧对称。玉米胚由腹部顶端形成胚芽鞘和生长锥的情况与水稻胚的胚芽鞘(顶生子叶)和生长锥的形成相同,玉米的胚芽鞘也是顶生子叶,盾片则是侧生子叶。玉米异型子叶的由来在于胚体的背腹极性。第三、玉米胚的真实形态结构及胚胎发育时期的划分。玉米胚是一个胚轴,其顶部是具胚芽鞘的胚芽,中部是具侧生子叶(盾片)的胚轴,下部是具胚根鞘的胚根。盾片从背面到腹面包住整个胚轴系统,在胚的腹面上可见从盾片边缘衍生的左、右侧鳞的边缘相迭,只在接缝线的上、下端留下蝌蚪状的小孔,使胚芽鞘和胚根鞘的末端稍露出。胚胎发育分为4个时期：1．原胚期——从合子细胞分裂开始至分化背部与腹部为止;2．腹部迅速分化期;3．盾片快速生长期;4．侧鳞生长、胚套形成期。第四、获取垂盲于胚腹面正中央纵切面是正确认识玉米胚形态的关键。     

稻属植物胚的形态结构与二（异）型子叶

长久以来植物学界认定稻 (OryzasativaL .)是单子叶植物。作者从稻胚发育的研究中确认稻胚具二型子叶 ,并非单子叶。稻属其他种的胚胎形态与O .sativa是否相同 ?是否具二型子叶 ?根据扫描电子显微镜的观察结果 ,稻属 (Oryza) 2 2个种和亚种的胚的形态结构可以分为两种类型。O .sativa等 16个种胚具腹鳞和侧鳞 ,属第一类型 ;O .meyeriana (Zoll.etMor.exSteud .)Baill.ssp .tuberculataW .C .WuetY .G .Lu ,G .C .Wang等 6个种 (亚种 )胚缺腹鳞和侧鳞 ,属第二类型。O .sativa和O .meyerianassp .tuberculata的胚胎发育过程所出现的盾片原基、胚根鞘原基、胚芽鞘原基和生长锥均来自原胚 ,前二者发育成胚套 ,是外围子叶 ;胚芽鞘原基发育成围在生长锥外并盖住生长锥的空心的倒锥状胚芽鞘 ,是顶生子叶。第一类型与第二类型稻胚都具有二型子叶。第二类型稻胚在盾片原基发育过程中并不分化出腹鳞和侧鳞 ,因而造成第二类型稻胚缺腹鳞与侧鳞。稻的二型子叶源于原胚的背腹极性分化     

稻属植物胚的形态结构与二(异)型子叶

长久以来植物学界认定稻(Oryza sativa L.)是单子叶植物.作者从稻胚发育的研究中确认稻胚具二型子叶,并非单子叶.稻属其他种的胚胎形态与O.sativa是否相同?是否具二型子叶?根据扫描电子显微镜的观察结果,稻属(Oryza) 22个种和亚种的胚的形态结构可以分为两种类型.O.sativa等16个种胚具腹鳞和侧鳞,属第一类型;O. meyeriana (Zoll. et Mor. ex Steud.) Baill.ssp. tuberculata W. C. Wu et Y. G. Lu, G. C. Wang等6个种(亚种)胚缺腹鳞和侧鳞,属第二类型.O.sativa和O. meyeriana ssp. tuberculata的胚胎发育过程所出现的盾片原基、胚根鞘原基、胚芽鞘原基和生长锥均来自原胚,前二者发育成胚套,是外围子叶;胚芽鞘原基发育成围在生长锥外并盖住生长锥的空心的倒锥状胚芽鞘,是顶生子叶.第一类型与第二类型稻胚都具有二型子叶.第二类型稻胚在盾片原基发育过程中并不分化出腹鳞和侧鳞,因而造成第二类型稻胚缺腹鳞与侧鳞.稻的二型子叶源于原胚的背腹极性分化.     

稻胚发育的三维形态研究兼论胚各部分的形态本质

运用扫描电镜及塑料半薄切片技术,从水稻（ＯｒｙｚａｓａｔｉｖａＬ．）授粉后２ｄ开始至种子成熟,追踪观察了稻胚的三维形态发育,根据结果,对胚各部分的形态本质提出一些新的见解。（１）授粉后２ｄ的胚由胚柄、胚基和胚体组成。胚基为胚柄和胚体之间的过渡区,呈喇叭状,胚基与胚柄不能等同。２ｄ的胚未出现器官分化,属原胚;但胚背腹分化明显,即存在背腹极性。（２）授粉后第３至第５天幼胚的形态变化及器官分化至关重要。盾片和胚芽鞘在授粉后３ｄ的幼胚上同时出现,两者均直接由原胚分化,并非胚芽鞘从盾片发生。胚芽鞘原基经历这３ｄ的特殊形态演变,形成空心倒锥状的胚芽鞘,展现了禾本科特有的胚芽鞘的形态形成机理。３ｄ幼胚胚根的原形成层、基本分生组织及根冠分化;４ｄ幼胚小丘状生长锥形成,胚根的原表皮分化,茎根轴形成;５ｄ幼胚胚芽、胚轴与胚根初步形成。（３）稻胚具有二型子叶,胚套是胚的外围子叶,盾片是此子叶的一个主要部分（侧生子叶）,胚芽鞘是顶生子叶。     

Oxygen Dependence of Elongation Growth and Oxygen Uptake in Maize Coleoptiles

Auxin-mediated elongation growth of maize coleoptile segments is inhibited by reducing the O₂ concentration in the incubation medium to GT 100 μmol . 1^?1. The half-maximal elongation rate is reached at 40 μmol . 1^?1 O₂, i.e. about two orders of magnitude higher than with mitochondrial respiration. O₂ uptake of the segments measured under similar conditions with an O₂ electrode shows a very similar dependence on O₂ concentration. Auxin increases O₂ uptake by 5–10% when it induces growth. About 40% of the O₂ uptake is insensitive to inhibition by KCN. Auxin has no effect on O₂ uptake in the presence of KCN. The possibility that auxin-mediated elongation growth depends on a KCN-sensitive oxidative process, other than cytochrome c oxidase-catalyzed respiration, is discussed.     

Reorientation of Microfibrils and Microtubules at the Outer Epidermal Wall of Maize Coleoptiles During Auxin-Mediated Growth

Auxin-mediated elongation growth of isolated subapical coleoptile segments of maize (Zea mays L.) is controlled by the extensibility of the outer cell wall of the outer epidermis (Kutschera et al., 1987). Here we investigate the hypothesis that auxin controls the extensibility of this wall by changing the orientation of newly deposited microfibrils through a corresponding change in the orientation of cortical microtubules. On the basis of electron micrographs it is shown that cessation of growth after removal of the endogenous source of auxin is correlated with a relative increase of longitudinally orientated microfibrils and microtubules at the inner wall surface. Conversely, reinduction of growth by exogenous auxin is correlated with a relative increase of transversely orientated microfibrils and microtubules at the inner wall surface. These changes can be detected 30–60 min after the removal and addition of auxin, respectively. The functional significance of directional changes of newly desposited wall microfibrils for the control of elongation growth is discussed.     

Extracellular matrix surface network of embryogenic units of friable maize callus contains arabinogalactan-proteins recognized by monoclonal antibody JIM4

Embryogenic units of friable maize callus are formed as globular or oblong packets of tightly associated meristematic cells. These units are surrounded by conspicuous cell walls visible in light microscopy after staining with basic fuchsin. Transmission electron microscopy revealed that embryogenic cells are rich in endoplasmic reticulum, polysomes and small protein bodies, and that the outermost layer of their cell walls is composed of fibrillar material. Electron microscopy has also shown that this material covers the surface of embryogenic cells as a distinct layer which we denote as extracellular matrix surface network (ECMSN). Employing histochemical staining with β-glucosyl Yariv phenylglycoside, we localized arabinogalactan-proteins (AGPs) to the outer cell walls of embryogenic units including ECMSN. The most prominent staining was found in cell-cell junction domains. Large non-embryogenic callus cells were not stained with this AGP-specific dye. Immunofluorescence and silver-enhanced immunogold labelling using monoclonal antibody JIM4 has shown that the ECMSN of embryogenic cells is equipped with JIM4 epitope, while non-embryogenic callus cells are devoid of this epitope. We propose that some specific AGPs of the ECMSN might be relevant for cell-cell adhesion and recognition of embryogenic cells during early embryogenic stages, and that the JIM4 antibody can serve as an early marker of embryogenic competence in maize callus culture. Received: 13 March 1998 / Revision received: 6 June 1998 / Accepted: 1 July 1998     

Comparative analysis of callus formation and regeneration on cultured immature maize embryos of the inbred lines A188 and A632

Induction, maintenance, differentiation and embryogenic capacity of callus obtained from immature embryos by culture on induction medium, proliferation medium, maturation medium and regeneration medium, respectively, were compared for two inbred lines of maize, i.e. A188 and A632. The callus of inbred line A188 was embryogenic and maintained embryogenic capacity for at least 1 year. Immature embryos of inbred line A632 formed callus that was not embryogenic. It only produced roots. When sucrose was replaced by sorbitol to induce or improve embryogenesis, again only A188 formed embryogenic callus. Subculture of this callus, however, allowed 4 week intervals in stead of 2 week intervals without loss of embryogenic capacity. When A188 was pollinated with A632 pollen, embryogenic callus was obtained from cultured immature "F₁" embryos, showing that embryogenic capacity was inherited, maternally. The callus did not differ from the embryogenic callus generated on selfed A188 embryos. When A632 was pollinated with A188 pollen, embryogenic callus was obtained too, showing that embryogenic capacity was also inherited paternally, though the embryogenic capacity diminished quickly, and the stability of the callus was lower than in the reciprocal cross. This revised version was published online in June 2006 with corrections to the Cover Date.     

Induction of elongation in maize coleoptiles by hexachloroiridate and its interrelation with auxin and fusicoccin action

The demonstration of an auxin-stimulated NADH-oxidase in the plasma membrane (Brightman et al. 1988. Plant Physiol. 86: 1264–1269) has led to the suggestion that the plasma membrane redox system is involved in the mechanism of auxin action. To evaluate the relevance of this concept in vivo, the influence of micromolar concentrations of hexachloroiridate (IV), an impermeable electron acceptor for the plant plasma membrane redox system, on elongation growth of excised, abraded maize coleoptile ( Zea mays L. cv. Golden Bantam) segments was studied. It was found that the substance induced a rapid growth response if the experiment was carried out in an unbuffered solution. This effect was entirely prevented by a 2 m M phosphate buffer. Nevertheless, the acid-growth-theory does not seem sufficient to explain this effect, since proton extrusion is induced without a lag, whereas increased growth rates commence after a lag phase of 40 min.
If growth is stimulated by a pretreatment with fusicoccin or auxin, hexachloroiridate IV transiently inhibits growth. The kinetics of the response are then determined by the concentrations of hexachloroiridate and auxin or fusicoccin. These results are compatible with the view that the plasma membrane redox system is somehow involved in the control of elongation growth.     

Gravitropic microtubule reorientation can be uncoupled from growth

The causal relationship between gravitropic growth responses and microtubule reorientation has been studied. Growth and microtubule reorientation have been uncoupled during the gravitropic response of maize (Zea mays L.) coleoptiles. Microtubule orientation and growth were measured under three different conditions: (i) a gravitropic stimulation where the growth response was allowed to be expressed (intact seedlings were displaced from the vertical position by 90°), (ii) a gravitropic stimulation where the growth response was suppressed (coleoptiles were attached to microscope slides and kept in a horizontal position), (iii) suppression of growth in the absence of gravitropic stimulation (coleoptiles were attached to microscope slides and kept in a vertical position). It was found that (i) gravitropic stimulation can induce a microtubular reorientation from transverse to longitudinal in the upper (slower growing) flank of the coleoptile, and an inhibition of growth; (ii) the reorientation of microtubules precedes the inhibition of growth; (iii) the gravitropic response of microtubules is weaker, not elevated, when the inhibition of growth is artificially enhanced by attaching the coleoptiles to a slide; and (iv) artificial inhibition of growth in the absence of gravitropic stimulation cannot induce a microtubular response. Thus, the extent of microtubule reorientation is not correlated with the extent of growth inhibition. Moreover, these findings demonstrate that microtubules do not reorient passively after growth changes, but actively in response to gravitropic stimulation. Received: 23 November 1999 / Accepted: 10 May 2000