白鲜营养器官解剖结构及其与白鲜碱的积累关系

应用植物解剖学、组织化学及植物化学方法对白鲜营养器官根、茎、叶的结构及其生物碱的积累进行了研究。结果显示:(1)白鲜根的次生结构以及茎和叶的结构类似一般双子叶植物;白鲜多年生根主要由周皮、次生韧皮部、维管形成层以及次生木质部组成,根次生韧皮部中可见大量的淀粉、草酸钙簇晶、韧皮纤维以及油细胞;茎由表皮、皮层、维管组织和髓组成;叶由表皮、栅栏组织、海绵组织和叶脉组成;在茎和叶初生韧皮部的位置均分布有韧皮纤维,在叶表皮上分布有头状腺毛和非腺毛;在茎和叶紧贴表皮处分布有分泌囊。(2)组织化学分析结果显示:在白鲜多年生根中,生物碱类物质主要分布在周皮、次生韧皮部、维管形成层和木薄壁细胞中;在茎中,生物碱主要分布在表皮、皮层、韧皮部、木薄壁细胞及髓周围薄壁细胞中;在叶中,生物碱主要分布在表皮细胞、叶肉组织和维管组织的薄壁细胞;此外在分泌囊和头状腺毛中亦含有生物碱类物质。(3)植物化学结果显示,秦岭产白鲜根皮/白鲜皮、根木质部、茎和叶中白鲜碱含量分别为0.041%、0.012%、0.004%和0.002%,其中木质部中白鲜碱含量和其他部分地区白鲜皮中白鲜碱含量类似。研究表明,在秦岭产白鲜营养器官中,除根皮/白鲜皮外,在根木质部亦含有大量的白鲜碱,且在茎和叶中亦含有一定的白鲜碱,具有潜在的开发利用价值。     

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。     

Distribution and frequency of plasmodesmata in relation to photoassimilate pathways and phloem loading in the barley leaf

Large, intermediate, and small bundles and contiguous tissues of the leaf blade of Hordeum tvulgare L. ‘Morex’ were examined with the transmission electron microscope to determine their cellular composition and the distribution and frequency of the plasmodesmata between the various cell combinations. Plasmodesmata are abundant at the mesophyll/parenchymatous bundle sheath, parenchymatous bundle sheath/mestome sheath, and mestome sheath/vascular parenchyma cell interfaces. Within the bundles, plasmodesmata are also abundant between vascular parenchyma cells, which occupy most of the interface between the sieve tube-companion cell complexes and the mestome sheath. Other vascular parenchyma cells commonly separate the thick-walled sieve tubes from the sieve tube-companion cell complexes. Plasmodesmatal frequencies between all remaining cell combinations of the vascular tissues are very low, even between the thin-walled sieve tubes and their associated companion cells. Both the sieve tube-companion cell complexes and the thick-walled sieve tubes, which lack companion cells, are virtually isolated symplastically from the rest of the leaf. Data on plamodesmatal frequency between protophloem sieve tubes and other cell types in intermediate and large bundles indicate that they (and their associated companion cells, when present) are also isolated symplastically from the rest of the leaf. Collectively, these data indicate that both phloem loading and unloading in the barley leaf involve apoplastic mechanisms.     

Degradation of the S. frugiperda peritrophic matrix by an inducible maize cysteine protease

A unique 33-kDa cysteine protease (Mir1-CP) rapidly accumulates at the feeding site in the whorls of maize (Zea mays L.) lines that are resistant to herbivory by Spodoptera frugiperda and other lepidopteran species. When larvae were reared on resistant plants, larval growth was reduced due to impaired nutrient utilization. Scanning electron microscopy (SEM) indicated that the peritrophic matrix (PM) was damaged when larvae fed on resistant plants or transgenic maize callus expressing Mir1-CP. To directly determine the effects of Mir1-CP on the PM in vitro, dissected PMs were treated with purified, recombinant Mir1-CP and the movement of Blue Dextran 2000 across the PM was measured. Mir1-CP completely permeabilized the PM and the time required to reach full permeability was inversely proportional to the concentration of Mir1-CP. Inclusion of E64, a specific cysteine protease inhibitor prevented the damage. The lumen side of the PM was more vulnerable to Mir1-CP attack than the epithelial side. Mir1-CP damaged the PM at pH values as high as 8.5 and more actively permeabilized the PM than equivalent concentrations of the cysteine proteases papain, bromelain and ficin. The effect of Mir1-CP on the PMs of Helicoverpa zea, Danaus plexippus, Ostrinia nubilalis, Periplaneta americana and Tenebrio molitor also was tested, but the greatest effect was on the S. frugiperda PM. These results demonstrate that the insect-inducible Mir1-CP directly damages the PM in vitro and is critical to insect defense in maize.     

DEVELOPMENT AND STRUCTURE OF THE VASCULAR CONNECTION BETWEEN THE PRIMARY AND SECONDARY ROOT OF GLYCINE MAX (L.) MERR.

The primary vascular connection between primary and secondary root of Glycine max (L.) Merr. was derived from stelar parenchyma and pericycle. Inner stelar parenchyma, associated with the parent metaxylem and outer stelar parenchyma adjacent to the pericycle, were resonsible for the histogenesis of the primary xylem connection. Acropetal maturation of the diarch xylem connection occurred after the lateral root emerged from the parent root. Development of tetrarchy occurred distal to the diarch xylem connection. The concentric primary phloem connection was derived from the pericycle and outer stelar parenchyma. Acropetal maturation of the primary phloem connection occurred prior to lateral root emergence from the parent root. Secondary growth quickly augmented the primary vascular connection. A substantial amount of mature secondary xylem formed prior to maturation of the secondary phloem. The structure of the primary and secondary vascular connections is described.     

Rice sucrose transporter1 (OsSUT1) up‐regulation in xylem parenchyma is caused by aphid feeding on rice leaf blade vascular bundles

The role of the sucrose transporter OsSUT1 in assimilate retrieval via the xylem, as a result of damage to and leakage from punctured phloem was examined after rusty plum aphid (Hysteroneura setariae, Thomas) infestation on leaves from 3‐week‐old rice (Oryza sativa L. cv Nipponbare) plants. Leaves were examined over a 1‐ to 10‐day infestation time course, using a combination of gene expression and β‐glucuronidase (GUS) reporter gene analyses. qPCR and Western blot analyses revealed differential expression of OsSUT1 during aphid infestation. Wide‐field fluorescence microscopy was used to confirm the expression of OsSUT1‐promoter::GUS reporter gene in vascular parenchyma associated with xylem elements, as well as in companion cells associated with phloem sieve tubes of large, intermediate and small vascular bundles within the leaf blade, in regions where the aphids had settled and were feeding. Of great interest was up‐regulation of OsSUT1 expression associated with the xylem parenchyma cells, abutting the metaxylem vessels, which confirmed that OsSUT1 was not only involved in loading of sugars into the phloem under normal physiological conditions, but was apparently involved in the retrieval of sucrose leaked into the xylem conduits, which occurred as a direct result of aphid feeding, probing and puncturing of vascular bundles. The up‐regulation of OsSUT1 in xylem vascular parenchyma thus provides evidence in support of the location within the xylem parenchyma cells of an efficient mechanism to ensure sucrose recovery after loss to the apoplast (xylem) after aphid‐related feeding damage and its transfer back to the symplast (phloem) in O. sativa leaves.     

草苁蓉瘤状体的形态解剖学研究

本文详细地观察了草苁蓉瘤状体的形态与结构,结果如下：草苁蓉瘤状体白色或深褐色,大小不等,最大直径约３０ｍｍ幼小时为球形,长大后为夫规则的块状;其解剖学特点是：瘤状体与寄主根连接的部分形成呼器,大量的薄壁细胞和螺纹状管胞伸入木质与根的木质部相连。吸器的真心皮部也由筛管伴胞及薄壁细胞与根的韧皮部相连,两者均缺乏纤维。     

DEVELOPMENT AND STRUCTURE OF THE VASCULAR CONNECTION BETWEEN THE PRIMARY AND LATERAL ROOT OF LYCOPERSICON ESCULENTUM

The primary xylem connection between the diarch parent root and the diarch lateral root was derived from the pericycle and stelar parenchyma. Early in lateral root development stelar parenchyma that was positioned between the parent xylem and the primordium divided transversely. These transverse divisions produced a plate of cells, most of which subsequently differentiated into vessel element connectors. After emergence of the lateral root, xylem maturation began in the stelar vessel element connectors and maturation proceeded acropetally into the lateral root. Protoxylem of the lateral root was connected to the metaxylem of the parent root via stelar vessel element connectors. The circular phloem connection was pericyclic in origin. Axial phloem connections which vascularized the lateral root were established with sieve tube elements of both parent phloem poles. Maturation of the phloem connection occurred prior to lateral root emergence. Transaxial phloem, positioned in arches above and below the lateral root vascular cylinder, was derived from the pericycle; and each arch consisted of three to four sieve tube elements. No transfer cells were found in the transaxial phloem.     

Expression patterns of vascular-specific promoters RolC and Sh in transgenic potatoes and their use in engineering PLRV-resistant plants

The expression patterns of GUS fusion constructs driven by the Agrobacterium rhizogenes RolC and the maize Sh (Shrunken; sucrose synthase-1) promoters were examined in transgenic potatoes (cv. Atlantic). RolC drove high-level gene expression in phloem tissue, bundle sheath cells and vascular parenchyma, but not in xylem or non-vascular tissues. Sh expression was exclusively confined to phloem tissue. Potato leafroll luteovirus (PLRV) replicates only in phloem tissues, and we show that when RolC is used to drive expression of the PLRV coat protein gene, virus-resistant lines can be obtained. In contrast, no significant resistance was observed when the Sh promoter was used.     

Histochemical localization of nucleoside triphosphatase activity in assimilate conducting plant tissue

Summary For the histochemical localization of nucleoside triphosphatases at the electron microscopic level, prefixed tissues were incubated with lead nitrate in addition to substrate (GOMORI reaction). While ATP and UTP as substrates gave electron-dense reaction products at the plasmalemma of sieve tubes, companion cells and phloem parenchyma cells, and at plasmodesmata in primary pitfields, AMP gave reaction products only at the tonoplast of parenchyma cells. Since electron-dense deposits also occur in cell walls and vacuoles, energy dispersive X-ray microanalysis was used to distinguish between lead deposits and lead-phosphate deposits. The latter were restricted to the symplast. Among the three plant species used, the leaf bundle phloem ofHordeum distichon showed ATPase activity largely restricted to the phloem cells, except for the thickwalled sieve tubes. Some activity also bordered the chloroplasts of the bundle sheath cells. In the C₄ plantGomphrena globosa, ATPase and UTPase activities appeared to be the greater in phloem parenchyma cells than in sieve tubes. In the phloem of youngMonstera deliciosa roots, ATPase occurred not only at the plasmalemma of sieve tubes, but also around sieve-tube plastids. When compared with AMP as substrate, it appears that nucleoside triphosphates are the natural substrates of the enzyme(s) in the plasmalemma of sieve tubes and phloem parenchyma cells.     

Histochemical IAA-oxidase localization in the shoot of wheat

A histochemical method for the determination of IAA-oxidase has been used in sections of various aerial parts of winter wheat plants. High IAA-oxidase activity was localized in the cell walls of sclerenchyma near the periphery of the stem, in the vascular bundle sheath of sclerenchyma and in xylem, both in the stem and in the leaf. The cell wall—bound IAA-oxidase activity therefore appeared in lignifying tissues. The staining was very weak or absent in the cell walls of parenchyma tissues and phloem. The positive reaction of the cytosol at the bulbous ends of guard cells and in the leaf primordia is presumed to be due to cytosolic IAA-oxidase. These results are discussed in relation to peroxidase localization and to our previousin vitro studies.     

六盘山鸡爪大黄蒽醌类化合物积累特征的研究

采用多种组织化学方法研究了六盘山鸡爪大黄营养器官中蒽醌类化合物的积累特征.结果显示:蒽醌类化合物在根中分布于周皮的木栓层和栓内层、次生维管组织的维管射线和根中央的部分木薄壁细胞内,且维管射线是根中贮藏和积累蒽醌类化合物的主要组织;在根茎中分布于周皮的木栓层和栓内层、次生维管组织的形成层和维管射线,以及髓的异常维管束射线中,且维管射线是根茎中贮藏和积累蒽醌类化合物的主要组织;在茎中主要分布于表皮、近表皮皮层和维管束的维管束鞘及其薄壁细胞,大型和小型维管束之间和周围的部分薄壁细胞,以及髓射线中有不同程度的分布;在叶中主要积累在叶柄的表皮、叶柄和大叶脉的部分基本组织、维管束的部分薄壁细胞等部位.结果表明,六盘山鸡爪大黄的根和根茎是蒽醌类化合物贮藏和积累的主要器官,维管射线是其贮藏和积累的主要组织,而且各营养器官中蒽醌类化合物积累的数量与植物各相关器官组织的发育程度、细胞中含淀粉粒的多少存在着一定的相关性.     

珊瑚菜植株分泌道发育和分布的解剖学观察

利用植物解剖学方法对珊瑚菜（Glehnia littoralis Fr．Schmidt ex Miq．）体内分泌道的发育和分布进行了观察。结果表明,珊瑚菜的分泌道有分枝,为溶生型,由1层分泌细胞围绕腔道而成。珊瑚菜叶片的分泌道发育较早,在幼叶阶段即发育成形。在根的次生韧皮部、根状茎的皮层和靠近初生木质部的髓部、叶脉的薄壁组织、叶柄维管束周围和厚角组织内侧的薄壁组织、花序轴正对维管束的皮层薄壁组织中以及果实的果壁维管束内外侧的薄壁组织中均分布有分泌道,分泌道在珊瑚菜体内分布广泛。     

The development of phloem anastomoses between vascular bundles and their role in xylem regeneration after wounding in Cucurbita and Dahlia

The differentiation of phloem anastomoses linking the longitudinal vascular bundles has been studied in stem internodes of Cucurbita maxima Duchesne, C. pepo L. and Dahlia pinnata Cav. These anastomoses comprise naturally occurring regenerative sieve tubes which redifferentiate from interfascicular parenchyma cells in the young internodes. In all three species, severing a vascular bundle in a young internode resulted in regeneration of xylem to form a curved by-pass immediately around the wound. The numerous phloem anastomoses in these young internodes were not involved in this process, the regenerated vessels originating from interfascicular parenchyma alone. Conversely, in mature internodes of Dahlia, the regenerated vessels originated from initials of the interfascicular cambia, and their phloem anastomoses did not influence the pattern of xylogenesis. On the other hand, in old internodes of Cucurbita, in which an interfascicular cambium was not yet developed, the parenchyma cells between the bundles had lost the ability to redifferentiate into vessel elements, and instead, regenerated vessels were produced in the phloem anastomoses. Thus, the wounded region of the vascular bundle was not bypassed via the shortest, curved pathway, but by more circuitous routes further away from the wound. Some of the regenerated vessels produced in the phloem anastomoses were extremely wide, and presumably efficient conductors of water. It is proposed that the dense network of phloem anastomoses developed during evolution as a mechanism of adaptation to possible damage in mature internodes by providing flexible alternative pathways for efficient xylem regeneration in plants with limited or no interfascicular cambium.This paper is dedicated to the memory of the late Isaac Blachmann (deceased 19 November 1995), father-in-law of the senior author, for encouragement and advice throughout the yearsThis research was supported by an International Scientific Exchange Award to R.A. from the Israel Academy of Sciences and The Royal Society.     

Perturbation of polyamine catabolism can strongly affect root development and xylem differentiation

Spermidine (Spd) treatment inhibited root cell elongation, promoted deposition of phenolics in cell walls of rhizodermis, xylem elements, and vascular parenchyma, and resulted in a higher number of cells resting in G(1) and G(2) phases in the maize (Zea mays) primary root apex. Furthermore, Spd treatment induced nuclear condensation and DNA fragmentation as well as precocious differentiation and cell death in both early metaxylem and late metaxylem precursors. Treatment with either N-prenylagmatine, a selective inhibitor of polyamine oxidase (PAO) enzyme activity, or N,N(1)-dimethylthiourea, a hydrogen peroxide (H(2)O(2)) scavenger, reverted Spd-induced autofluorescence intensification, DNA fragmentation, inhibition of root cell elongation, as well as reduction of percentage of nuclei in S phase. Transmission electron microscopy showed that N-prenylagmatine inhibited the differentiation of the secondary wall of early and late metaxylem elements, and xylem parenchymal cells. Moreover, although root growth and xylem differentiation in antisense PAO tobacco (Nicotiana tabacum) plants were unaltered, overexpression of maize PAO (S-ZmPAO) as well as down-regulation of the gene encoding S-adenosyl-l-methionine decarboxylase via RNAi in tobacco plants promoted vascular cell differentiation and induced programmed cell death in root cap cells. Furthermore, following Spd treatment in maize and ZmPAO overexpression in tobacco, the in vivo H(2)O(2) production was enhanced in xylem tissues. Overall, our results suggest that, after Spd supply or PAO overexpression, H(2)O(2) derived from polyamine catabolism behaves as a signal for secondary wall deposition and for induction of developmental programmed cell death.     

Cell differentiation in the longitudinal veins and formation of commissural veins in rice (Oryza sativa) and maize (Zea mays)

Vascular development is a central theme in plant science. However, little is known about the mechanism of vascular development in monocotyledons (compared with dicotyledons). Therefore, we investigated sequential processes of differentiation into various different vascular cells by carrying out detailed observations using serial sections of the bases of developing leaves of rice and maize. The developmental process of the longitudinal vascular bundles was divided into six stages in rice and five stages in maize. The initiation of differentiation into procambial progenitor cells forming the commissural vein arose in a circular layer cell that was adjacent to both a metaxylem vessel and one or a few phloem cells in stage V longitudinal vascular bundles. In most cases the differentiation of ground meristem cells into procambial progenitor cells extended in one direction, toward the next longitudinal vascular bundle, and subsequent periclinal divisions and further differentiation produced a vessel element, two companion cells and a sieve element to form a commissural vein. These results suggest the presence of an intercellular signal(s) that induces differentiation of the circular layer cell and the ground meristem cells into procambial progenitor cells, forming a commissural vein sequentially.     

Sites of accumulation in excised Phloem and vascular tissues

Excised pieces of vascular bundle and phloem tissue were allowed to accumulate radioactive phosphate and sulfate, and were then sectioned and autoradiographed so as to detect the sites of accumulation. Special methods were needed to prevent any diffusion of the radioisotope. Some autoradiographs obtained are presented. In excised celery vascular bundles, the most radioactive area and hence the most actively accumulating tissue was the young secondary phloem at the sides of the bundle. In intact plants, the same tissue was the most active in translocating. In excised apple phloem there was some variation in behavior, but again the young secondary phloem was generally the most actively accumulating tissue. Accumulation activities of individual cells in the phloem and vascular tissue were compared. It appeared that all cell types, ray, phloem and xylem parenchyma, cambial cells and sieve tubes, accumulated at least 5 times more actively than did the cortical parenchyma cells. The sieve tubes were among the most actively accumulating cells present, accumulating 20 times more actively than the cortical parenchyma cells. It is concluded that accumulation processes have a primary role to play in the mechanism of phloem transport.