Anatomical Studies of Haustorium Ontogeny and the Remarkable Mode of Penetration of the Haustorium in Nuytsiafloribunda (Labill.) R. Br.

The development and structure of secondary haustoria of Nuytsia floribunda are described and compared with other Santalalean haustoria. After establishing contact with the host root, cortical folds of the haustorium grow around the root in separate directions and fuse forming a ring around it. At an early stage of development, meristematic tissue differentiates in the interior proximal part of the haustorium. Zones of collapsed layers are present in the outer cortical region. Subsequently, in the proximal part, two vascular cores, two lysigenous cavities and extensive masses of sclerenchyma develop prior to penetration of the host root. The sclerenchymatous cells form a characteristic structure, described as the sclerenchyma prong. During penetration the intrusive part of the haustorium reaches not only the host xylem but continues growing downwards until it entirely splits the host root. Comparable to a guillotine, the sclerenchyma prong is directly involved in this remarkable process. The sclerenchyma prong finally lies in the distal part of the haustorium. Following this mechanical slicing of the host root, tube-like cells of the intrusive part actively penetrate the host xylem in an axial direction.     

Development of Stem Nodules in a Tropical Forage Legume, Aeschynomene afraspera

Rhizobial infection occurred on the stem of Aeschynomene afrasperaat the site of emergence of adventitious root primordia. Rhizobiainvaded cortical cells at the base of the root primordium. Thefirst infected cell enlarged and collapsed after rhizobia hadmultiplied in large numbers. At this time, a meristematic zonewas initiated some distance beneath the first infected cell.Rhizobial penetration into the deeper cortex was by progressivecollapse of infected cells towards the meristematic zone; rhizobiaentering the cortical cells by invagination of the host cellwall. At the entry point, rhizobia were embedded in digitatecell wall material. These infection structures were restricted,always originating from the cell wall of an adjacent infectedcell. Soon after infection, the cell collapsed progressivelyforming infection strand-like structures which developed upto the meristematic zone. When infection had reached the meristematiczone, invaded host cells ceased to collapse but divided repeatedlyto form the nodule. Key words: Aeschynomene afraspera, stem nodulation     

Initiation, Development and Structure of the Primary Haustorim in Striga gesnerioides (Scrophulariaceae)

A light-microscopic study is reported on the initiation, establishmentand structure of the primary haustorium of Striga gesnerioideson the host, cowpea (Vigna unguiculata). The radicular apexof the germinated parasite seed dissolves its way through thehost root cortex to the stele. Thus, it is converted into aprimary haustorium. Some of the haustorial front-line cellsin contact with the host endodermis penetrate into the steleand make contact with the xylem vessels. Differentiation ofthese haustorial cells into xylem vessels occurs and extendsbackwards through the median axial region of the haustorialtract in the host cortex to connect with the conductive xylemof the radicle outside the host root. Subsequently the parasite'splumule develops into a leafy shoot. On penetrating the steleof the host, the haustorium stimulates cell division in thehost pericycle whose triggered proliferation together with expansionof the parasite haustorial tissues result in the formation ofa large, tuberous primary haustorium. At various points of thehost-parasite interface, differentiation of xylem elements occurs,presumably maximizing nutrient transfer from host to parasite.In spite of this, many proliferated host cells at the interfaceremain apparently meristematic showing densely-stained cytoplasmand prominent nuclei.     

Development and Structure of the Haustorium of the Parasite Rhamphicarpa fistulosa (Scrophulariaceae)

Rhamphicarpa fistulosa (Hochst.) Benth. (Scrophulariaceae), a parasite of African cereals, develops secondary haustoria which penetrate the roots of the host plant. Light and electron microscopy have been used to study the structure and development of haustoria in this species, which, until now, have not been well characterized. Haustoria are initiated in the hypodermis of the parasite roots. A meristematic strand is developed between the parasite root stele and the host-parasite interface. From this strand, cells differentiate into xylem elements after penetration of the host root. Xylem differentiation follows an acropetal pattern. Mature haustoria are characterized by a continuous xylem bridge between water conducting elements of parasite and host. A detailed study of the hostparasite interface revealed the presence of collapsed and compressed host cells at the lateral interface (between parasite cells and host cortex), whereas the central interface between parasite cells and the host stele is almost devoid of host cell remnants. Implications of these observations for the penetration mechanisms are discussed.     

The structure and development of the haustorium in parasitic Scrophulariaceae*

The structure and development of roots and haustoria in 37 species of parasitic Scrophulariaceae was studied using light microscopy. The mature haustorium consists of two regions: the swollen “body” and the parent root, which resembles non-haustorial roots in structure. The body arises from the parent root and is composed of an epidermis, cortex, central region of xylem (the vascular core), a region of parenchyma (the central parenchymatous core), and the portion of the haustorium contained in the host tissue (the endophyte). The xylem of the vascular core is composed predominately of vessel elements. The central parenchymatous core is composed of parenchyma and col-lenchyma. Vessels extend from the vascular core through the central parenchymatous core to the endophyte. The endophyte is composed of parenchyma cells and vessel elements. No phloem is present in the body of the haustorium. Early stages in the development of the haustorium are exogenous. Initial periclinal divisions in the epidermis or outer cortex are followed by hypertrophy of cortical parenchyma. These events are followed by development of the vascular core from the pericycle, attachment of haustorium to the host by a specialized layer of cementing cells or root hairs, and penetration of the host by dissolution of host cells.     

檀香吸器维管组织的个体发育

为了解檀香吸器维管组织的发育过程,采用激光共聚焦显微镜、光学显微镜和透射电镜观察檀香吸器维管组织的个体发育。结果表明,檀香维管组织的分化分为两个时期:入侵前和入侵后。吸器维管组织发育始于盘状吸器时期,起源于吸器基部具有分生能力的细胞,后分为两束。侵入前无向顶的分化,处于吸器基部。侵入后随吸管深入寄主根与寄主根维管束连通,形成具有吸收功能的维管组织。成熟吸器维管组织呈倒烧瓶结构,仅处于吸器烧瓶核心两边,由木质部组成而无韧皮部。檀香的吸器维管组织发育有两个因素诱导,一个是遗传因素,另一个为寄主。这些为檀香半寄生性特性研究提供了形态解剖学基础。     

An anatomical study of the haustoria of Rhinanthus minor attached to roots of different hosts

Roots of a range of potential hosts responded differently when Rhinanthus minor attempted to form haustoria. Roots of Fabaceae show the weakest reaction as apart from slight lignification, no reaction was observed at the interface between the endophyte and the cortical tissue of the host root. Grass roots react with strong lignification of all cells within the stele with the exception of a small number of phloem cells whilst the endodermis fully enters the tertiary stage. In the case of Phleum bertolonii the cortical cells also become lignified. The lignification is even observed in the host root tissue in a distance of about 1 mm from the haustorium (both apically and basipetally). In the case of Leucanthemum vulgare, strong suberisation can be observed in the cell walls of the interface between endophyte (tip of the sucker) and host. Plantago lanceolata exhibits the strongest reactions against the haustorial tissues. Cells of the interface between the endophyte and the host cortex are completely destroyed, as well as a few cell layers outside the central xylem cylinder, even in some distance from the haustorium. Thus, host xylem is completely isolated from the haustorium in this case. Extraction of sap from xylem vessels is likely to be drastically impaired in such a situation.     

THE HOST-PARASITE INTERFACE OF ALBUGO CANDIDA ON RAPHANUS SATIVUS

The fine structure of the intercellular hyphae of the obligate parasite Albugo candida infecting radish does not differ markedly from that described previously for cells of Peronospora manshurica. The stalked, capitate haustoria do not contain nuclei and are packed with mitochondria and lomasomes. The fungal plasma membrane and cell wall are continuous from the intercellular hypha throughout the haustorium except that there is no evidence of fungal cell wall around a portion of the haustorial stalk proximal to the haustorial head. Within the vacuolate host mesophyll cell, the haustorium is always surrounded by host plasma membrane and with at least a thin layer of host cytoplasm. The host cell wall invaginates at the point of haustorial penetration to form a short sheath around the region of penetration, but normally there is no host cell wall around the balance of the haustorium. About 1% of the haustoria observed were necrotic, and these were invariably walled-off completely from host cytoplasm by host cell wall. An amorphous, moderately electron-dense encapsulation lies between the haustorium proper and the host plasma membrane and extends into the penetration region between the sheath and the fungal cell wall. Invaded host cells contain more ribosomal-rich ground cytoplasm than uninfected cells. Glandular-like systems of tubules and connecting vesicles are often numerous in host cytoplasm in the vicinity of haustorial heads. These tubules open into the encapsulation, their limiting unit membranes being continuous with the host plasma membrane. We suggest that these represent a secretory mechanism of the host specifically induced by the parasite.     

Morphology and Anatomy of the Haustorium of the Root Hemiparasite Olax phyllanthi (Olacaceae), with Special Reference to the Haustorial Interface

Gross morphology and internal structure of haustoria of Olaxphyllanthi are described in parasitism with a range of hosts,including roots of woody and herbaceous dicotyledons and certainmonocotyledons, and occasional instances of autoparasitism andhaustorial formation on monocotyledon rhizomes. Successful penetrationto xylem occurs on virtually all hosts across broad diameters,ages and anatomies of host root, but anatomical impedimentsto haustorial establishment and penetration are recorded forcertain host taxa. Each haustorium is a comparatively simpleand ephemeral structure. Its developing sucker (endophytic regionof the haustorium) spreads laterally around the surface of thehost xylem, yet never completely encircles the host stele. Damageto hosts is minimal and secondary thickening (of hosts) continueson the side of a host root opposite to a haustorium. The haustorialsucker lacks phloem and its interface with host xylem is comprisedalmost entirely (more than 98.7%) of parenchyma. The few terminatingtracheids at an interface lie in very close proximity to oroccasionally directly against exposed xylem vessels, but lumento lumen continuity between tracheary elements of the partnersis not achieved. Three dimensional reconstructions based onserial transverse sectioning indicate that well defined filesof tracheids connect back from an interface to the core of graniferoustracheary elements in the external body of the haustorium, andthence to the xylem of the parent parasite root. The findingsare discussed in relation to existing studies on haustorialanatomy. Root parasite, Olacaceae, haustorial anatomy, host specificity     

The infection process ofFusarium oxysporum in cotton root tips

Summary Conidia ofFusarium oxysporum f. sp.vasinfectum started to germinate on the roots of cotton (Gossypium barbadense L.) 6 h after inoculation and formed a compact mycelium covering the root surface. 18 h later, penetration hyphae branched off and infected the root. The number of penetration hyphae increased with the number of conidia used for inoculation. The optimal temperature for penetration was between 28 and 30 °C. The highest numbers of penetration hyphae were found in the meristematic zone, 40 percent less in the elongation and root hair zones, and none in the lateral root zone. The fine structure of the infection process was studied in protodermal cells of the meristematic zone and in rhizodermal cells of the elongation zone. The penetration hyphae were well preserved after freeze substitution and showed a Golgi equivalent consisting of three populations of smooth cisternae. Plant reactions were found already during fungal growth on the root surface. In the meristematic zone, a thickening of the plant cell wall due to an apposition of dark and lightly staining material below the hyphae occurred. This wall apposition increased in size around the hypha invading the plant cell and led to the formation of a prominent wall apposition with finger-like projections into the host cytoplasm. In the elongation zone, the deposits around the penetration hypha appeared less thick and the dark inclusions were less pronounced. High pressure freezing of infected cells revealed, thatF. oxysporum penetrates and grows within the host cells without inducing damages such as plasmolysis, cell degeneration or even host necrosis. We suggest thatF. oxysporum has an endophytic or biotrophic phase during colonization of the root tips.Abbreviation Ph penetration hyphae     

Green tissue within the haustorium of the dwarf mistletoe Korthalsella (Viscaceae). An ultrastructural comparison between chloroplasts of sucker and aerial stem tissues

Summary Korthalsella (Viscaceae) is a dwarf mistletoe attached to its host branch by a single haustorium. Plants are leafless with flattened or cylindrical stems that function in photosynthesis. When a fresh haustorium is cut the sucker within the host appears bright green. Transmission electron microscopy reveals that this greening is due to chloroplasts, but that their organization differs from those of the aerial stem. The three representatives of Korthalsella endemic to New Zealand were the main species investigated. In the stem, chloroplasts have short stacks of cylindrical grana interconnected by stroma thylakoids typical of normal chloroplasts. Sucker chloroplasts have a more variable organization, with most containing extensive granal stacks and poorly differentiated stroma thylakoids. These granal thylakoids exhibit extensive partitions formed by appression of adjacent membranes. Some sucker plastids also approach etioplasts in having a prominent prolamellar body from which radiate thylakoids with short partitions. Sucker chloroplasts usually contain a few large starch grains, plastoglobuli, and sometimes also a stroma centre. The extensive granal thylakoids in sucker chloroplasts of Korthalsella resemble that found in certain shade plants and tissue grown under low light conditions. Sucker chloroplasts probably have a low level of photosynthesis. This activity might provide a local source of osmotically active material used to assist transport between host and parasite.     

Root histogenesis from tobacco thin cell layers

Summary Internode stem expiants ofNicotiana tabacum cv. Samsun, consisting of eight cell layers: epidermis, subepidermal chlorenchyma, collenchyma and cortical parenchyma (i.e., thin cell layers), were cultured under conditions inducing rhizogenesis. The aim was to investigate the histological sequence of adventitious root formation in this system. The earliest cytological events in culture (12 h) were nucleolar extrusions and amitotic nuclear divisions. Though not restricted to a specific cell layer, the two phenomena were more frequent in the subepidermal chlorenchyma, and characterized the first phases (12-96 h) of cell proliferation mainly occurring in this layer. Amitoses were followed by the formation of thin walls within the original cells, resulting in the formation of intracellular clusters. These subepidermal clusters were separated by enlarged cells of the parent tissue, whose nuclei showed nucleolar extrusion. At day 3 the first mitoses were observed in cells having abundant starch inclusions. Amitotic divisions also continued, but less frequently. The increasing frequency of mitoses in the subepidermal chlorenchyma (day 4), as well as in the two underlying collenchymatous layers, contributed to the growth of the superficial clusters, in which small clumps of meristematic cells were formed; these, later (day 9), gave rise to root domes. The 5th cell layer remained undivided for a relatively long time (two weeks). The 6th and 7th layers proliferated mitotically later (from day 8 onwards) than the superficial layers and formed root domes following the same histological sequence. Wound callus, generated by the innermost layer, increased markedly in the last two weeks of culture and concomitantly formed vascular clumps surrounded by meristematic layers; these produced root primordia which were frequently anomalous (day 26–27). Regardless of its origin (i.e., superficial or deep layers of the expiant, or wound callus cells), root tip formation was always preceded by the differentiation of a sheath of starch-containing cells, from which the root cap developed.Abbreviations LS longitudinal section - S.E. standard error - TVS transverse section     

ANATOMY AND FINE STRUCTURE OF THE MISTLETOE HAUSTORIUM (PHTHIRUSA PYRIFOLIA). I. DEVELOPMENT OF THE YOUNG HAUSTORIUM

The anatomy and fine structure of the young primary haustorium of Phthirusa pyrifolia (H.B.K.) Eichl. were studied before penetration into the host. The simple internal organization (epidermis, hypodermis, and core parenchyma) which characterizes the radicular disc at germination becomes extremely complex, especially at the distal end of the disc during haustorial development. The epidermis in the area of contact with the host surface develops into an intricate cell zone consisting of lobed and tubular portions. The tubular portions consist of finger-like projections that entwine and form bulbous tips at the contact surface. The tubular portions have unusual wall thickenings while the bulbous tips have exceedingly thin distal walls which possibly break, releasing their contents onto the host's surface. The collapsed layers characteristic of Santalalean haustoria seem to be a result of internal pressures caused by division and expansion of epidermal cells and core parenchyma. Various unusual ultrastructural features are described from the hypodermis, core parenchyma, and contact zone. Particularly striking, but yet unidentified, is a fibrillar material which often completely fills the cells of the core parenchyma in later stages of development.     

Ultrastructure of graniferous tracheary elements in the haustorium of Exocarpus bidwillii, a root hemi-parasite of the Santalaceae.

The xylem in the body of the haustorium of E. bidwillii has the shape of an inverted conical flask with the expanded portion being known as the vascular core. The tracheary elements of the vascular core are notable for the occurrence of numerous granules within their lumina and the presence of mostly imperforate walls. Elsewhere in the haustorium graniferous tracheary elements are absent and the cells are usually ordinary vessel elements. Thin sections for transmission electron microscopy, post-stained in potassium permanganate, show that the secondary wall thickenings of the graniferous tracheary elements consist of eccentric layers in which the microfibrils of each successive layer run alternately longitudinally and transversely. The granules of the tracheary elements average 2 micrometer in diameter and consist of a homogeneous matrix which shows a fine fibrillar structure on high resolution. The granules are naked and mostly remain as separate structures within the lumen of the cell, but occasionally they fuse into small groups or irregular masses. In some cells the granules become transformed into fibrillar material that disperses throughout the lumen. This dispersed material may accumulate in vessels of the interrupted zone proximal to the vascular core. Occasionally, the granules also change into compacted amorphous masses that adhere to the walls of the cell. Ultrastructural cytochemistry confirms that the granules are protein and not starch as was originally believed for the Santalaceae. The function of the vascular core and its graniferous tracheary elements is discussed and we suggest that it might help regulate the pressure and flow of xylem sap entering the parasite from the host. Graniferous tracheary elements in the Santalaceae and in root parasites of the Serophulariaceae are compared and it is concluded that they represent examples of convergent evolution.     

Light and Electron Microscopy of the Compatible Interaction Between Arabidopsis and the Downy Mildew Pathogen Peronospora parasitica

In this study, we focused on compatible interactions between Peronospora parasitica isolate Emoy‐2 and wild‐type (Oy‐0) and mutant (Ws‐eds1) Arabidopsis thaliana accessions by using light and transmission electron microscopy (TEM). Light microscopy of compatible interactions revealed that conidia germinated and penetrated through the anticlinal cell walls of two epidermal cells. Rapid spreading of the hyphal growth with formation of numerous haustoria within the mesophyll cells was subsequently followed by profuse sporulation in the absence of host cell necrosis on both wild‐type and mutant accessions. TEM observations revealed that coenocytic intercellular hyphae ramified and spread intercellularly throughout the host tissue forming several haustoria in host mesophyll cells. Intracellular haustoria were lobed with the diameter of 6–7 μm. Each haustorium was connected to intercellular hyphae in the absence of apparent haustorial neck. The cytoplasm of the haustorium included the organelles characteristic of the pathogen. Callose‐like deposits were frequently observed at sites of penetration around the proximal region of the haustorial neck. Apart from a few callose ensheatments, no obvious response was observed in host cells following formation of haustoria. Most of mesophyll cells contained normal haustoria and the host cytoplasm displayed a high degree of structural integrity. Absence of host cell wall alteration and cell death in penetrated host cell of both accessions suggest that the pathogen exerts considerable control over basic cellular processes and in this respect, response to this biotroph oomycete differs considerably from responses to other pathogens such as necrotrophs.     

Untersuchungen zur Anfälligkeit und Resistenz von Kopfsalat (Lactuca sativa) gegen falschen Mehltau (Bremia lactucae): II. Licht- und elektronenmikroskopische Untersuchungen zur Morphologie des Wirt-Parasit-Verhältnisses

Investigations on the susceptibility and resistance of head lettuce (Lactuca sativa) to downy mildew (Bremia lactucae) II. Light and electron microscopic examinations of the host-parasite interface Infected leaves of lettuce varieties susceptible and incompletely resistant to Bremia lactucae were observed by light and electron microscopy. Primary infection structures in the epidermal cells as well as intercellular hyphae with the adjacent haustoria could be seen by differential interference contrast microscopy. The haustoria in host cells of susceptible varieties collapsed before degeneration of the invaded host cell. On the contrary, host cells of incompletely resistant varieties died before the haustoria in these cells showed any sign of degeneration. Electron microscopic investigations confirmed the observations with light microscopy. In incompletely resistant varieties, an electron transparent sheath enveloped the haustorium. In the sheath fragments of membranes are localized. These membrane particles as seen by using the goniometer in electron microscopic work were flat faced. The sheath material consists of transformed host cell wall material and involves fragments of the host plasmalemma as well as fragments of the unit membrane separating the sheath from extrahaustorial matrix. The sheath has an important role as a special filter to prevent the passage of nutrients from the host cell into the haustorium. Thus the incomplete resistance is based not only on an impeded penetration of the parasite into the epidermal cells and their hypersensitive reactions in case of a successful penetration but also on hypersensitivity of mesophyll cells which does not necessarily lead to death of the parasite but does impede the absorption of nutrients.     

A comparative light microscopic study of two types of root nodule bacteria dispersal into the bacteroid zone cells

Root nodule bacterial dispersal into the cells and formation of bacteroid zone cells of root nodules have been observed in several species of Leguminosae. Two different types of bacterial dispersal were recognized comparable to those briefly mentioned by a few authors. Cell division type: From the very beginning of nodule development there exist two different kinds of cell in the meristematic region. In bacteria-containing cells the bacteria are distributed into two new cells with host cell division, the bacteria being located outside the spindle region during mitosis. In those cells not containing bacteria, amyloplasts develop in the cytoplasm. These cells also divide mitotically, increasing the numbers of amyloplast-containing cells. These two different cell populations compose the bacteroid zone. This type was seen inSophora flavescens andCytisus scoparius. Infection thread type: In the transitional zone from the nodule meristem to the bacteroid zone, some cells are penetrated by infection threads and others are not. Infected cells become bacteroid-filled cells, no mitosis taking place after penetration of the infection thread. Uninfected cells become amyloplast-containing cells, increasing their numbers by mitosis. These two cell populations compose the bacteroid zone. This type was seen inVicia sativa, V. faba, Trifolium repens andAstragalus sinicus.     

The Initiation of the Secondary Haustorium in Alectra vogelii Benth

Anatomical studies were carried out on initiation of the secondaryhaustorium in Alectra vogelii, a root parasite of leguminouscrops in Nigeria. In both the normal and self-haustorium, theformation of the haustorial initial on the parasite root soonafter initial contact between the host and parasite roots isfollowed by the penetration of the host root by the haustorium.Specialized penetrating cells (intrusive cells) at the haustorialfront prise apart and loosen the host root cortical cells, whichlater become digested. Through the same processes, a few ofthese columnar intrusive cells at the haustorial front piercethe endodermis to make contact with the xylem of the host root.Thereafter, a true conductive bridge consisting of short, isodiametric,reticulate vessel elements is established between the parasiteand host roots through the secondary haustorium. No pholem tissuewas observed in the connection. There is a close similaritybetween the mode of initiation of the secondary haustorium ofAlectra vogelii and that previously described for its primaryhaustorium. Alectra vogelii Benth, haustorium, self-haustorium, root parasite, hemiparasitism, Vigna unguiculata, Arachis hypogaea     

Epidermal derivatives as xylem elements and transfer cells: a study of the host-parasite interface in two species of Triphysaria (Scrophulariaceae)

Summary Haustoria ofTriphysaria pusilla andT. versicolor subsp.faucibarbata from a natural habitat were analysed by light and electron microscopy. The keel-shaped edge of the secondary haustorium generally splits the epidermis and cortex of the host root parallel to the root axis, and penetrates to the host vascular tissue. Anticlinally elongated epidermal cells of the haustorium constitute most of the host/parasite interface. Some of these epidermal cells are divided by oblique cell walls. Some of their oblique daughter cells as well as some undivided epidermal cells differentiate into xylem elements. Single epidermal cells occasionally intrude into the vascular tissue of the host and individual host cells can be invaded. The surface area of the plasmalemma in parasitic parenchymatous interface cells is increased by the differentiation of wall labyrinths characteristic of transfer cells and by the development of membrane-lined cytoplasmic tubules or flattened sacs which become embedded in the partly lignified interface cell-wall. Mycorrhizal fungal hyphae enter the xylem bridge in some haustoria. Implications of these observations for the function of the haustorium are discussed.     

Changes of Plastids During Xylem Differentiation in Barley Root

Plastids (eoplasts) are present in meristematic cells of prospectivecentral metaxylem in the barley root. Starch starts to be formedin plastids precisely after the cessation of mitotic activityand at the beginning of endomitotic growth. During secondarywall formation, the starch is gradually lost. Cavities are formedin plastids and signs of plastid degeneration are present fromthis stage of cell development. However, some intact globularplastids without starch are present until shortly before thefinal step of ontogeny, i.e. total destruction of protoplast. Hordeum distichum L., root, xylem, plastids, endomitotic growth, starch