木立芦荟叶内芦荟素的超微细胞化学研究

使用醋酸铅溶液对木立芦荟叶的药用成分芦荟素进行细胞化学定位,在透射电镜下探讨芦荟素产生、转运和贮藏的过程。结果表明:芦荟素由同化薄壁组织产生,质体的类囊体为其合成部位。通过质体膜形成的小泡转移到周围的内质网,以后内质网小泡与质膜融合:或质体小泡直接与质膜融合。通过胞吐作用将芦荟素释放到质膜外,经质外体途径到达维管束的鞘细胞。在鞘细胞中, 芦荟素经内质网小泡转移至内切向壁,由胞间连丝运输到芦荟素细胞的细胞质,最终贮藏在芦荟素细胞的液泡中。     

木立芦荟不同叶龄叶的解剖学和组织化学及其植物化学研究

应用植物解剖学、组织化学、荧光显微观察与植物化学技术相结合,研究了木立芦荟不同叶龄叶的解剖结构、叶绿素和类胡萝卜的含量、芦荟素的含量及其合成和贮藏结构的特点。结果表明:芦荟素由同化薄壁组织产生,叶绿体的基质为其合成场所。芦荟素细胞可能是芦荟素早期贮存的场所,随着芦荟素细胞的逐渐萎缩老化,维管束鞘细胞成为贮藏芦荟素的代替场所。同一植株的叶随着叶龄的增长,维管束的密度降低,芦荟素细胞占维管束横切面的百分比减小,同化薄壁组织细胞中的叶绿体逐步衰老、解体,芦荟素含量逐渐降低,但不同叶龄叶中叶绿体色素的含量与芦荟素含量间没有明显的相关性。     

六种芦荟叶的解剖结构及其与芦荟素含量的相关性

应用半薄切片法、高效液相色谱法(HPLC)和荧光显微镜研究了6种芦荟叶的结构、芦荟素的含量和储藏芦荟素的组织。结果表明,6种芦荟叶均由表皮、光合组织、储水组织和维管束组成,都表现出明显的旱生植物肉质叶的结构特征,表皮由一层扁平的细胞组成,其外壁加厚,并覆盖着厚的角质膜,气孔器凹陷,储水组织发达。6种芦荟叶的结构存在显著差异。木立芦荟(Aloe arborescens Mill．)和易变芦荟(A．mutabilis Pillans)的光合组织细胞呈长柱状,类似栅栏薄壁组织。中华芦荟(A．vera L.var．chinensis Berg．)、库拉索芦荟(A．vera L．)、皂叶芦荟(A．saponaria Hawer)和绿芦荟(A．greenii Bak．)则为等直径薄壁细胞。木立芦荟、中华芦荟、易变芦荟和库拉索芦荟的维管束中有大型薄壁细胞,皂叶芦荟和绿芦荟的维管束中无大型薄壁细胞。木立芦荟、易变芦荟和库拉索芦荟在光合组织和储水组织之间有一层不含叶绿体的小型薄壁细胞,包围着储水薄壁组织,称之为储水组织鞘。中华芦荟、皂叶芦荟和绿芦荟则没有储水组织鞘。HPLC测量结果表明,木立芦荟叶芦荟素含量最高,库拉索芦荟和易变芦荟次之,中华芦荟、皂叶芦荟和绿芦荟含量最低。荧光显微镜观察结果表明,在紫外光和蓝光下,黄色和黄绿色小球体仅存在于维管束的大型薄壁细胞、维管束鞘和储水组织鞘中,而光合组织和储水组织中没有黄色和黄绿色小球体。因此,维管束中大型薄壁细胞、维管束鞘和储水组织鞘是芦荟素等蒽醌类物质的储藏场所。综上所述,芦荟素含量与维管束的大型薄壁细胞、维管束鞘和储水组织鞘的情况密切相关。     

六种芦荟叶的解剖结构及其与芦荟素含量的相关性

应用半薄切片法、高效液相色谱法(HPLC)和荧光显微镜研究了6种芦荟叶的结构、芦荟素的含量和储藏芦荟素的组织.结果表明,6种芦荟叶均由表皮、光合组织、储水组织和维管束组成,都表现出明显的旱生植物肉质叶的结构特征,表皮由一层扁平的细胞组成,其外壁加厚,并覆盖着厚的角质膜,气孔器凹陷,储水组织发达.6 种芦荟叶的结构存在显著差异.木立芦荟(Aloe arborescens Mill.)和易变芦荟(A. mutabilis Pillans)的光合组织细胞呈长柱状,类似栅栏薄壁组织.中华芦荟(A. vera L. var. chinensis Berg.)、库拉索芦荟(A. vera L.)、皂叶芦荟(A. saponaria Hawer)和绿芦荟(A. greenii Bak.)则为等直径薄壁细胞.木立芦荟、中华芦荟、易变芦荟和库拉索芦荟的维管束中有大型薄壁细胞,皂叶芦荟和绿芦荟的维管束中无大型薄壁细胞.木立芦荟、易变芦荟和库拉索芦荟在光合组织和储水组织之间有一层不含叶绿体的小型薄壁细胞,包围着储水薄壁组织,称之为储水组织鞘.中华芦荟、皂叶芦荟和绿芦荟则没有储水组织鞘.HPLC测量结果表明,木立芦荟叶芦荟素含量最高,库拉索芦荟和易变芦荟次之,中华芦荟、皂叶芦荟和绿芦荟含量最低.荧光显微镜观察结果表明,在紫外光和蓝光下,黄色和黄绿色小球体仅存在于维管束的大型薄壁细胞、维管束鞘和储水组织鞘中,而光合组织和储水组织中没有黄色和黄绿色小球体.因此,维管束中大型薄壁细胞、维管束鞘和储水组织鞘是芦荟素等蒽醌类物质的储藏场所.综上所述,芦荟素含量与维管束的大型薄壁细胞、维管束鞘和储水组织鞘的情况密切相关.     

甘蔗叶不同部位ATP酶活性细胞化学定位

甘蔗叶片,叶鞘和肥厚带韧皮部 ATP 酶活性定位于筛管、伴胞的质膜、内质网和某些伴胞细胞基质、小囊泡和发育成熟的液泡上;叶片韧皮部薄壁细胞、厚壁细胞和厚壁通道细胞质膜及小囊泡中亦显示有 ATP 水解产物;维管束鞘细咆与厚壁细胞或厚壁通道细胞所构成的细胞间隙上也存在有 ATP 酶活性反应产物沉淀。甘蔗叶片大、中、小三种维管束,从小维管束到大维管束,面向细胞间隙的细胞表面上的 ATP 酶活性逐渐增强,而维管束鞘细胞质膜上的 ATP 酶活性则趋于减弱;同一维管束内则以韧皮部细胞的 ATP 酶活性最强。维管束鞘细胞与叶肉细胞之间存在很多的胞间连丝,并表现出高的 ATP 酶活性。讨论了 ATP 酶活性的分布状态与叶肉细胞的光合产物向韧皮部运输的关系。     

芦荟叶内芦荟素细胞的发育和蒽醌类物质的积累

应用石蜡切片、半薄切片、组织化学和荧光显微镜观察相结合的方法研究了木立芦荟叶内芦荟素细胞的发生、发育以及其蒽醌类物质的积累过程。结果表明,在叶内原形成层束分化成维管束初期,原形成层束外侧的一层细胞发育成维管束鞘。原生韧皮部筛管产生时,其外方尚保留1-2层原形成层细胞,当后生韧皮部和木质部开始分化时,此层细胞分裂。在后生韧皮部和木质部发育成熟过程中,这些细胞体积逐渐增大,并液泡化,发育成为大型薄壁细胞(芦荟素细胞),位于筛管外侧。据此,芦荟叶维管束内的大型薄壁细胞的来源与韧皮部相同,属于特化的韧皮部薄壁组织细胞。用醋酸铅处理过的上述材料的切片观察表明,芦荟素细胞在细胞体积增大,并液泡化时,在液泡内出现蒽醌类物质沉淀物,在成熟细胞的大液泡中充满沉淀物,此时,在荧光显微镜下芦荟素细胞发出桔黄色荧光。可见,此种芦荟素细胞是芦荟叶内蒽醌类物质的主要储存场所。     

芦荟叶内芦荟素细胞的发育和蒽醌类物质的积累

应用石蜡切片、半薄切片、组织化学和荧光显微镜观察相结合的方法研究了木立芦荟叶内芦荟素细胞的发生、发育以及其蒽醌类物质的积累过程。结果表明,在叶内原形成层束分化成维管束初期,原形成层束外侧的一层细胞发育成维管束鞘。原生韧皮部筛管产生时,其外方尚保留1—2层原形成层细胞,当后生韧皮部和木质部开始分化时,此层细胞分裂。在后生韧皮部和木质部发育成熟过程中,这些细胞体积逐渐增大,并液泡化,发育成为大型薄壁细胞(芦荟素细胞),位于筛管外侧。据此,芦荟叶维管束内的大型薄壁细胞的来源与韧皮部相同,属于特化的韧皮部薄壁组织细胞。用醋酸铅处理过的上述材料的切片观察表明,芦荟素细胞在细胞体积增大,并液泡化时,在液泡内出现蒽醌类物质沉淀物,在成熟细胞的大液泡中充满沉淀物,此时,在荧光显微镜下芦荟素细胞发出桔黄色荧光。可见,此种芦荟素细胞是芦荟叶内蒽醌类物质的主要储存场所。     

芦荟维管束的结构与芦荟素积累的相关性

应用半薄切片、组织化学、荧光显微镜观察和薄层层析 (TLC)相结合的方法研究了中华芦荟 (Aloeve-ra L.var.chinensis)、木立芦荟 (Aloe arborescens)叶和茎内维管束的结构及其与芦荟素积累的关系。结果表明 ,木立芦荟叶内维管束和中华芦荟叶内外轮的维管束中有大型韧皮薄壁细胞 ,而木立芦荟茎和中华芦荟叶中内轮维管束无大型韧皮薄壁细胞。组织化学结果表明 ,用醋酸铅处理过的上述材料 ,大型韧皮薄壁细胞内出现沉淀物 ;在荧光显微镜下经蓝光激发 ,大型韧皮薄壁细胞发出桔黄色荧光 ,都显示出芦荟素反应。薄层层析(TLC)结果证明 ,木立芦荟和中华芦荟叶含有大型韧皮薄壁细胞的维管束都含芦荟素 ,而木立芦荟茎及中华芦荟叶中内轮维管束都不含芦荟素。为此 ,维管束中的大型韧皮薄壁细胞与芦荟素的积累密切相关 ,维管束中是否有大型韧皮薄壁细胞可作为判断是否含有芦荟素的解剖学指标。     

Anatomical structure and distribution of secondary metabolites as a peripheral defence strategy in Aloe hereroensis leaves

Eight leaves from four different plants of Aloe hereroensis and 18 leaf parts of each leaf were tested by anatomic, fluorescence microscopy and TLC methods. Four phenolic secondary metabolites, homonataloin and three isomers of aloeresin, were found in the leaves. The highest content of these metabolites was found in the top third of a leaf and along the leaf margins. In the margins, the content of the four secondary metabolites in the adaxial was higher than in the abaxial direction. In the centre parts of the leaves, the metabolic content of the abaxial parts was higher than in the adaxial parts. The results indicate that homonataloin mainly accumulates in the inner bundle sheath cells (IBSC). The three isomers of aloeresin appear in the outer bundle sheath and in the boundary cells between the chlorenchyma and water-storage tissues. The density of the vascular bundles, the area ratio of the chlorenchyma to the tested counterpart, and the area ratio of the IBSC to a whole bundle are important structural factors to determine the differences in the content of these four secondary metabolites in all the leaf parts. The distribution according to the rosette leaf arrangement and the existence of the 'cocktail' of four phenolic secondary metabolites indicate a peripheral defence strategy of this plant. © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 138 , 107–116.     

Secretory tissues in the East African shrubby aloes

Examination of Aloe leaf sections revealed the presence of three types of cells at the phloem pole of the vascular bundles, aloin cells, outer bundle sheath cells and fibres. Three species contain fibres alone and produce a sparse exudate with few components staining purple with Fast Blue B on thin-layer chromatograms. The majority of species have aloin cells of various sizes and in the tetraploid species these produce a copious exudate containing anthraquinone and chromone derivatives. It is suggested that the aloin cells act as storage tissue and that the compounds are synthesized in the surrounding layer of cells of smaller diameter, many of which can be seen to contain globules of unknown constitution. Anatomical observations support the idea that the shrubby tetraploid Aloe species are derived from a form similar to the diploid Aloe morijensis. Two forms of this species have been described, one with fibres only and few exudate components and another with both fibres and aloin cells and an exudate containing compounds of the tetraploid species. A line of diploid plants similar in anatomy and chemistry to the first form is represented by A. fibrosa and A. babatiensis while a line of tetraploids with affinities to the second form is represented by A. nyeriensis, A. cheranganiensis, A. elgonica, A. dawei and A.yavellana.     

Aloin Protects Skin Fibroblasts from Heat Stress-Induced Oxidative Stress Damage by Regulating the Oxidative Defense System

Oxidative stress is commonly involved in the pathogenesis of skin damage induced by environmental factors, such as heat stress. Skin fibroblasts are responsible for the connective tissue regeneration and the skin recovery from injury. Aloin, a bioactive compound in Aloe vera, has been reported to have various pharmacological activities, such as anti-inflammatory effects. The aim of this study was to investigate the protective effect of aloin against heat stress-mediated oxidative stress in human skin fibroblast Hs68 cells. Hs68 cells were first incubated at 43°C for 30 min to mimic heat stress. The study was further examined if aloin has any effect on heat stress-induced oxidative stress. We found that aloin protected Hs68 cells against heat stress-induced damage, as assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and lactate dehydrogenase assay. Aloin protected Hs68 cells by regulating reactive oxygen species production and increasing the levels of glutathione, cytosolic and mitochondrial superoxide dismutase. Aloin also prevented the elevation of thiobarbituric acid reactive substances and the reduction of 8-OH-dG induced by heat stress. These results indicated that aloin protected human skin fibroblasts from heat stress-induced oxidative stress damage by regulating the oxidative defense system.     

Development and ultrastructure of the megagametophyte in Passiflora caerulea L. (Passifloraceae)

Megasporogenesis and megagametogenesis of Passiflora caerulea L. were studied using light and transmission electron microscopy. The archesporial tissue is generally formed by one cell. The megaspore mother cell gives rise to a linear tetrad of megaspores. The chalazal megaspore is the functional one, and originates a Polygonum -type female gametophyte. The antipodals are ephemeral. Abundant starch is found in the nucellar cells, specially the ones adjacent to the megagametophyte. The two synergids show ultrastructural differences, involving the filiform apparatus, the nucleolus and the endoplasmic reticulum; these differences suggest a functional differentiation, probably related to the reception of the pollen tube. This is the first report in angiosperms of substantial morphological differences between the two synergids.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society , 2003, 142 , 73–81.     

Anthraquinones and β-polysaccharides content and distribution in Aloe plants grown under different light intensities

The curative and therapeutic effects of Aloe plants have mostly been ascribed to anthraquinones such as aloin, and to some characteristic β-polysaccharides. Although the actual concentration of these bioactives in Aloe plants has not yet been fully clarified, it was expected that plant species, age and growth conditions would play an important role. The aim of this work was to investigate the relationship between species, light intensity and the content of bioactives in Aloe arborescens Mill. and Aloe vera (L.) Burm.f.Aloin was determined by liquid chromatography tandem mass spectrometry: Its concentration was higher in the leaves of younger plants and there was more in A. vera than in A. arborescens. The content of β-polysaccharides was determined colorimetrically after binding with Congo Red dye. The results were not affected by plant age, and concentrations were higher in A. vera than in A. arborescens.Finally, even though the type of tunnel (and therefore light spectrum) under which plants were grown seemed to have no effect on the content of bioactives, the plants grown under reduced light intensities had significantly lower aloin and β-polysaccharides concentrations.     

Comparative vegetative anatomy and systematics of the Oncidiinae (Maxillarieae, Orchidaceae)

Subtribe Oncidiinae comprises a vegetatively heterogeneous assemblage of species that has persistently been incapable of organization. Anatomy was considered to be a possible means to resolve the perplexity of relationships amongst the constituent taxa. The consistent occurrence of a foliar hypodermis, homogeneous mesophyll, conical silica bodies in stegmata, and ubiquitous fibre bundles in leaves provides a matrix for linking the taxa, as do the parenchymatous pith and O-thickened endodermal cell walls in roots. However, the strict consensus of the 40 genera studied was completely unresolved, suggesting that vegetative characters alone are insufficient to assess the relationships amongst these taxa, a conclusion also reached for the remainder of Maxillarieae.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 152 , 91–107.     

Biomechanics and development of rattans: what is special about Plectocomia himalayana Griff. (Calamoideae,Plectocomiinae)?

Mechanical and morphological studies of Plectocomia himalayana (subtribe Plectocomiinae) revealed characteristics that differ strongly from species of subtribe Calaminae ( Calamus and Daemonorops ). In species of Calaminae tested previously, the contribution of the leaf sheath drastically increases stiffness in juvenile axes and towards the apex of older plants. In P. himalayana the relative contribution of the leaf sheath to axis stiffness is less and leaf sheath senescence does not strongly reduce axial stiffness as observed in Calamus and Daemonorops . Natural aerial branching, only described in Korthalsia and Laccosperma among rattans, is common in P. himalayana . Aerial branching and adventitious roots occur frequently along old stems allowing autonomy of stems, following mechanical injury and promoting vegetative propagation. The climbing habit is known to have evolved at least twice within the Calamoideae. The results observed here suggest that climbing habits may differ in detail and that different 'climbing strategies' may have evolved within the subfamily Calamoideae resulting from: (1) variable stem flexibility, (2) the variable mechanical role of the leaf sheath ( Calamus–Daemonorops ) and (3) production of branches and aerial roots conferring a higher degree of architectural plasticity ( Plectocomia ).  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 151 , 83–91.     

Aloin enhances cisplatin antineoplastic activity in B16-F10 melanoma cells by transglutaminase-induced differentiation

Aloin, a natural anthracycline from aloe plant, is a hydroxyanthraquinone derivative shown to have antitumor properties. This study demonstrated that aloin exerted inhibition of cell proliferation, adhesion and invasion abilities of B16-F10 melanoma cells under non-cytotoxic concentrations. Furthermore, aloin induced melanoma cell differentiation through the enhancement of melanogenesis and transglutaminase activity. To improve the growth-inhibiting effect of anticancer agents, we found that the combined treatment of cells with aloin and low doses of cisplatin increases the antiproliferative activity of aloin. The results suggest that aloin possesses antineoplastic and antimetastatic properties, exerted likely through the induction of melanoma cell differentiation.     

Morphometric, AFLP and plastid microsatellite variation in populations of Scalesia divisa and S. incisa (Asteraceae) from the Galápagos Islands

Analysis of the stem vasculature of the American climbing palm Desmoncus reveals structural features differing significantly from the Old World rattan genus Calamus . Desmoncus has a more directly continuous vascular system but nevertheless shows a vessel distribution that makes for high hydraulic resistance in the axial xylem. Desmoncus is like Calamus in having a single very wide metaxylem vessel in each central axial bundle and is also without direct vascular contact between protoxylem and metaxylem tracheary elements. However, in Desmoncus the stem vascular bundle system resembles that in tree palms (as has been described in the model palm Rhapis excelsa ) in having a continuing axial bundle that branches from each outgoing leaf trace together with a large number of bridge connections between leaf traces and peripheral axial bundles. Resistance to axial water transport is, however, evident in the narrowness of the continuing metaxylem elements in the peripheral stem vascular region. Desmoncus has scalariform perforation plates with few thickening bars in the metaxylem vessels, unlike the simple perforation plates found in Calamus . Thus, Desmoncus shows only limited convergence in stem vascular architecture toward the extreme modifications found in Calamus . This is not unexpected since it is clear that the climbing habit evolved independently in the two genera.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society, 2003, 142 , 243−254.     

芦荟属植物叶内蒽醌类物质的组织化学定位研究

蒽醌类物质是芦荟属（Aloe)植物叶内贮存的重要次生代谢物质,主要是芦荟素,高那特芦荟素,芦荟苦素和芦荟宁等。根据蒽醌类物质的理化性质,用2％－5％NaOH和5％Pb(CH3COO)2溶液对材料进行处理,再辅以荧光和超薄切片观察等多种方法,对芦荟叶内的4种主要蒽醌类物质进行了组织化学定位研究。结果表明,蒽醌类物质在芦荟叶内的贮藏是多位点的。芦荟素和高那特芦荟素主要位于韧皮部的大型薄壁组织细胞之中;而芦荟苦素和芦荟宁主要位于维管束鞘细胞以及贮水组织与同化薄壁组织之间的1圈薄壁组织细胞之中,贮水组织细胞中不贮存蒽醌类物质。     

Plasmodesmatal frequency in relation to short-distance transport and phloem loading in leaves of barley (Hordeum vulgare). Phloem is not loaded directly from the symplast

We investigated the phloem loading pathway in barley, by determining plasmodesmatal frequencies at the electron microscope level for both intermediate and small blade bundles of mature barley leaves. Lucifer yellow was injected intercellularly into bundle sheath, vascular parenchyma, and thin-walled sieve tubes. Passage of this symplastically transported dye was monitored with an epifluorescence microscope under blue light. Low plasmodesmatal frequencies endarch to the bundle sheath cells are relatively low for most interfaces terminating at the thin- and thick-walled sieve tubes within this C₃ species. Lack of connections between vascular parenchyma and sieve tubes, and low frequencies (0.5% plasmodesmata per μm cell wall interface) of connections between vascular parenchyma and companion cells, as well as the very low frequency of pore-plasmodesmatal connections between companion cells and sieve tubes in small bundles (0.2% plasmodesmata per μm cell wall interface), suggest that the companion cell-sieve tube complex is symplastically isolated from other vascular parenchyma cells in small bundles. The degree of cellular connectivity and the potential isolation of the companion cell-sieve tube complex was determined electrophysiologically, using an electrometer coupled to microcapillary electrodes. The less negative cell potential (average –52 mV) from mesophyll to the vascular parenchyma cells contrasted sharply with the more negative potential (–122.5 mV) recorded for the companion cell-thin-walled sieve tube complex. Although intercellular injection of lucifer yellow clearly demonstrated rapid (0.75 μm s^-1) longitudinal and radial transport in the bundle sheath-vascular parenchyma complex, as well as from the bundle sheath through transverse veins to adjacent longitudinal veins, we were neither able to detect nor present unequivocal evidence in support of the symplastic connectivity of the sieve tubes to the vascular parenchyma. Injection of the companion cell-sieve tube complex, did not demonstrate backward connectivity to the bundle sheath. We conclude that the low plasmodesmatal frequencies, coupled with a two-domain electropotential zonation configuration, and the negative transport experiments using lucifer yellow, precludes symplastic phloem loading in barley leaves.     

Latitudinal and ecological correlates of morphological variation in Galaxias platei (Pisces, Galaxiidae) in Patagonia

We examined the morphological variability (23 morphometric traits) among individual Galaxias platei ( N  = 380) collected from 20 postglacial lakes in the southern Andes. The lakes were chosen to cover the latitudinal range of the species in Patagonia. Diet examined for a subset of these fish ( N  = 261) collected during the summer months (January through March), differed among river basins but there were no clear latitudinal trends in the consumption of any prey type. Diet may, however, have partially shaped morphology, as pelvic measures were negatively correlated with consumption of amphipods. Substantial differentiation among populations was observed, primarily in the shape and dimensions of the head, in caudal morphology and in fin length. Our results indicate that the morphology of G. platei varies with latitude, and may be related to risk of predation and diet.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 87 , 69–82.