Rhinanthus serotinus (Schönheit) Oborny (Scrophulariaceae): immunohistochemical and ultrastructural studies of endosperm chalazal haustorium development

Chalazal endosperm haustorium in Rhinanthus serotinus consists of a single large binucleate cell. It originates from the primary endosperm cell dividing transversely into two unequal cells: a smaller micropylar cell and a larger chalazal cell. The chalazal cell undergoes a single mitotic division, then lengthens significantly during development and functions as a chalazal endosperm haustorium. In this paper, immunofluorescent techniques, rhodamine phalloidin assay, and electron microscopy were used to examine the actin and tubulin cytoskeleton during the development of the chalazal haustorium. During the differentiation stage, numerous longitudinally oriented bundles of microfilaments ran along the axis of transvacuolar strands in haustorium. Microtubules formed intensely fluorescent areas near the nuclear envelope and also formed radial perinuclear microtubule arrays. In the fully differentiated haustorium cell, the actin cytoskeleton formed dense clusters of microfilaments on the chalazal and micropylar poles of the haustorium. Numerous microfilament bundles occurred near wall ingrowths on the chalazal wall. There were numerous clusters of microfilaments and microtubules around the huge lobed polytenic haustorial nuclei. The microfilaments were oriented longitudinally to the long axis of the haustorium cell and surrounded both nuclei. The microtubules formed radial perinuclear systems which were appeared to radiate from the surface of the nuclear envelope. The early stage of degeneration of the chalazal haustorium was accompanied by the degradation of microtubules and disruption of the parallel orientation of microtubules in the chalazal area of the cell. The degree of vacuolization increased, autophagous vacuoles appeared and the number of vesicles decreased.     

Influence of carbohydrates on the induction of haustoria of the cowpea rust fungusin vitro

Rust fungi are obligate plant parasites whose successful parasitism appears to depend on the formation of haustoria within living plant cells. To test the hypothesis that the lack of haustorium formation in the absence of a living cell reflects a need for a simple “signal” released during attempted invasion, various carbohydrates were applied toin vitro-formed infection structures ofUromyces vignae just prior to haustorial mother cell (HMC) formation. Up to 6% of HMCs formed haustoria after treatment with arabinose, mannose, xylose, sucrose, or xylan. Other monosaccharides, oligosaccharides, and polysaccharides were less or not effective. Sugar mixtures induced haustorium frequencies of up to 10%. Timing of the application of inducing chemicals was critical. The data indicate that haustorium formation does not require the presence of a living plant cell and may be triggered by plant products at the time of penetration peg development.     

不同抗病性小麦品种上白粉菌吸器发育超微结构研究

利用电镜技术对不同抗病性小麦品种上白粉菌吸器发育及相应寄主细胞变化的超微结构进行了研究,并对吸器的Ca2+－ATP酶活性及几丁质的分布进行了细胞化学定位分析。结果表明,小麦白粉菌（Blumeriagraminisf．sp.tritici）成熟吸器在内部结构上类似一个代谢活跃的菌丝细胞,有大量的线粒体和多聚核糖体;Ca2+－ATP酶主要被定位在寄主质膜及病菌核膜上;随吸器的不断发育,吸器外膜厚度增加,同时Ca2+-ATP酶活性增强。几丁质均匀地分布在吸器壁上,其含量随吸器的成熟而增加。在不同抗病性小麦品种上,吸器细胞核最先退化,然后是线粒体的液泡化和多聚核糖体的解聚。中抗寄主细胞内的吸器普遍退化较早,相当一部分在吸器中心体阶段已解体。此外,高感寄主表皮细胞与叶肉细胞之间有发达的胞间连丝;而且在吸器形成后,能比中抗寄主细胞更快地增殖和聚积大量与能量代谢、物质合成及分泌活动有关的细胞器。     

Occurrence of Endosperm Haustorium in Cannabis sativa L.: With thirteen Figures in the Text

The development and structure of the chalazal endosperm haustoriumin Cannabis sativa are described. The endosperm is nuclear anda haustorium is formed at the chalazal end. The latter remainsfree nuclear throughout. Enucleate vesicles appear in the upperpart of the endosperm but finally they merge with the cytoplasmof the haustorium. As the embryo reaches maturity it occupiesthe whole seed cavity, the haustorium collapses and the endospermpersists only as a thin layer.     

西瓜胚和胚乳的发育

应用显微技术对西瓜胚和胚乳的发育过程进行了观察并分析了西瓜胚珠败育的原因。西瓜胚发育属紫菀型。合子第一次分裂为不均等分裂 ,形成的基细胞体积明显较顶细胞大 ,两细胞均含有多个液泡。原胚发育过程中没有明显的胚柄。最外层的原胚细胞 ,与胚乳细胞相邻的壁上被胼胝质物质包围 ,且无外连丝存在 ;与胚囊壁相接的壁上无壁内突结构。胚的子叶体积增长的同时 ,子叶细胞内积累蛋白质和脂类物质 ,多糖物质的含量下降。胚乳发育属核型 ,在球形胚期开始自珠孔端向合点端细胞化 ,胚子叶分化出后开始自珠孔端向合点端退化。胚乳合点端在球形胚早期形成发达的胚乳吸器 ,开始呈游离核状态 ,后细胞化 ,在心型胚期之后退化。     

西瓜胚乳吸器的发育及ATP酶的超微细胞化学定位

报道了西瓜（Citrullus lanatus)胚乳吸器发育过程,并对胚乳吸器细胞中的ATP酶进行了超微细胞化学定位,球形胚早期,胚囊合点端的壁伸长发育成一管状胚乳吸器,进而吸器靠近乳本体端膨大为囊状,球形胚晚期吸器自珠孔端向合点端逐渐细胞化,胚分化出子叶时,胚乳吸器自合点端向珠孔端退化,在刚形成的胚乳吸器细胞中,ATP酶活性反应主要分布在细胞的核膜,内质网上,胞间连丝和吸器细胞壁内的小球状物上也有较强的ATP酶活性反应;在开始退化的吸器细胞中,核膜上的ATP酶性的反应减弱较早,内质网稍晚,进一步退化的胚乳吸器细胞中,ATP酶主要集中分布在细胞壁,细胞间隙内,核上几乎没有ATP酶性反应,内质网上仅有微弱的ATP酶反应。     

罗汉果胚、胚乳及胚乳吸器的发育

本文采用石蜡切片与酶解分离法对罗汉果Ｓｉｒａｉｔｉａｇｒｏｓｖｅｎｏｒｉ胚、胚乳及胚乳吸器的发育过程进行观察．ａ）罗汉果胚的发育是按Ｇｅｕｍｕｒｂａｎｕｍ的分裂程序进行的．属紫菀型．但在合子分裂成球胚过程中,胚芽原细胞分化明显．故属紫菀型的变异型。ｂ）胚乳发育属核型．在球形胚阶段,在合点端和珠孔端有发育的胚乳吸器形成并进行旺盛生长,最大长度达１４２０μｍ,心形胚期．吸器活动开始减退,合点端核型胚乳吸器转变成细胞型．由胚乳本体基部膨大细胞．充当补助吸器．ｃ）酶解分离法研究胚乳吸器的发生发育有较好的应用前景。     

Anatomy of the endophyte of Viscum album L. (Loranthaceae)

Anatomy of the endophyte of Viscum album L. (Loranthaceae). An anatomical investigation into the nature of the host-parasite interaction of V. album and several of its phanerogamic hosts using SEM and light microscopy was conducted. Three kinds of parasite cell (haustorial parenchyma cells, cells resembling transfer cells and haustorial tracheids) were identified at the host-parasite interface. The terms haustorial parenchyma and haustorial tracheid are defined. Haustorial tracheids were seen to have penetrated the walls of host vessel elements and it is suggested that V. album is able to establish on a wide range of hosts because of the anatomically plastic nature of its haustorium. The development of the haustorium depends to a large extent on the nature of the surrounding host tissues. Parasite-induced host abnormalities including hypertrophy, distorted xylem elements, vessel-wall penetration and tylosis-occluded vessels were observed. The macroanatomical features observed are discussed and interpreted by-proposing a new theory for the ontogenesis of the V. album haustorium. Cortical strands with 'chisel' and 'pencil' shaped apices were both found to be present at the same time on one plant and thus were not seasonally separated.     

The parasitic mechanism of the holostemparasitic plant Cuscuta

Cuscuta is a stem holoparasitic plant without leaves or roots, which develops a haustorium and sucks nutrients from host plants. The genus Cuscuta comprises about 200 species, many of which can cause severe problems for certain crops. The parasitic process in Cuscuta begins in finding and attaching to a host plant and then developing a haustorium. The process does not always require any chemical signal, but does require a light signal. Finding a host involves detecting the lower red light:far-red light ratio near a potential host plant by phytochrome. A contact signal is also necessary for haustorium induction. Apparently, cytokinin increase is downstream of the light and contact signal and is critical for haustorium induction. This pathway, however, appears to be slightly different from a standard pathway. The direct connection between Cuscuta and its host involves both the xylem and phloem, and mRNA and proteins can translocate. Several features indicate that Cuscuta is a useful model plant for parasite plant research as well as plant–plant interaction research. These include the simple anatomical structure and seedling development, no chemical requirement for haustorium induction, and the wide range of host plants.     

In Vitro Haustorium Development in Roots and Root Cultures of the Hemiparasitic Plant Triphysaria Versicolor

Parasitic plants in the Orobanchaceae invade host plant roots through root organs called haustoria. Parasite roots initiate haustorium development when exposed to specific secondary metabolites that are released into the rhizosphere by host plant roots. While molecular approaches are increasingly being taken to understand the genetic mechanism underlying these events, a limitation has been the lack of a transformation system for parasitic plants. Since the haustorium development occurs in roots of Orobanchaceae, root cultures may be suitable material for transient or stable transformation experiments. To this end, root cultures were obtained from explants, and subsequently calluses, from the hemiparasitic plant Triphysaria versicolor. The cultured roots retained their competence to form haustoria when exposed to host roots, host root exudates, or purified haustorium-inducing factors. The root culture haustoria invaded host roots and initiated a vascular continuity between the parasite and host roots. The ontogeny of haustoria development on root cultures was indistinguishable from that on seedlings roots. Root cultures should provide useful material for molecular studies of haustorium development.     

肉苁蓉寄生生长形态发育

肉苁蓉(Cistanche deserticola)是一种沙生根寄生濒危药用植物。利用光镜和电镜手段, 采用人工培养和诱导等方法详细观察并研究了肉苁蓉寄生生长过程中种子萌发、吸器产生以及植物体形态发育的过程。结果表明: (1)人工可以诱导肉苁蓉的种子萌发。肉苁蓉种胚具有明显的极性, 珠孔端细胞小于合点端, 珠孔端细胞分化、生长并产生白色的类胚根状结构。(2)有些化学物质可以诱导初生吸器的产生。用2, 6二甲氧基-对苯醌培养肉苁蓉24-26小时后, 其先端膨大并产生突起, 形成类似根毛状的结构, 即初生吸器。(3) 肉苁蓉属于主动寄生植物, 其在初生吸器与寄主幼根黏连后产生次生吸器。肉苁蓉与寄主梭梭(Haloxylon ammodendron)共培养, 其初生吸器主动与寄主幼根(0.1 mm左右)黏连, 穿过寄主根表皮和皮层后与维管束连通形成次生吸器, 肉苁蓉植物体分化、发育的基本组织形成。被寄生后的寄主根横向发育加快, 同时肉苁蓉植物体开始分化和发育。(4)肉苁蓉可以寄生在寄主幼根(0.1 mm左右)的任意部位。     

梨胶锈菌性孢子和锈孢子阶段吸器的超微结构研究

本文对梨胶锈菌性子期和锈子期菌丝吸器的形成方式、吸器及其与寄主细胞界面的超微结构进行了研究。观察结果表明：梨胶锈菌性子期和锈子期寄主胞间菌丝吸器的形成方式有两种：一种是由寄主胞间菌丝直接形成吸器;另一种是由寄主胞间菌丝先形成吸器母细胞,然后由吸器母细胞形成吸器。吸器在开始形成时只是一个乳头状的侵入楔,以后逐渐形成囊状、镰刀状、指状及其它不规则形状的吸器。多数吸器分化为颈和吸器主体两部分,在颈部及部分吸器主体外有一个由类似寄主细胞壁物质形成的领圈。吸器内部的超微结构与寄主胞间菌丝基本相同,但吸器壁比胞间菌丝或吸器母细胞的壁薄。吸器鞘的厚度随着吸器伸长膨大 而逐渐增厚。     

The Differentiation of Infection Structures as a Result of Recognition Events between some Biotrophic Parasites and their Hosts

Most uredospores of rust fungi develop infection structures in a typical pattern so that they can infect the host plant. The function of these infection structures is divided into the following three phases:

• 1 In the recognition phase, the germ tube recognizes the cuticle and the stoma. This process may occur independently from the host plant since copies of the cuticle induce similar reactions of the fungus. During fungal growth on the epidermis, unspecific stress responses of the plant are triggered.
• 2 In the signal phase, the fungal substomatal vesicle and infection hypha(e) contact the host cells within the leaf parenchyma. A signal from the host induces further development of the fungus. Haustorium mother cell differentiation is effected and haustorium formation is initiated. At the same time, the fungus suppresses the synthesis of stress metabolites by the plant.
• 3 In the parasitic phase, the fungus penetrates the host cell and complex interactions between host and parasite begin. A highly specialized interface around the haustorium develops presumably in order to allow a more efficient nutrient transfer from host to parasite. Eventual defence reactions of the plant, generally on the race-cultivar level, fail to be evoked or are suppressed in compatible combinations.

     

Inoculation with arbuscular mycorrhizal fungi suppresses initiation of haustoria in the root hemiparasite Pedicularis tricolor

Background and Aims

Plant parasitism and arbuscular mycorrhizal (AM) associations have many parallels and share a number of regulatory pathways. Despite a rapid increase in investigations addressing the roles of AM fungi in regulating interactions between parasitic plants and their hosts, few studies have tested the effect of AM fungi on the initiation and differentiation of haustoria, the parasite-specific structures exclusively responsible for host attachment and nutrient transfer. In this study, we tested the influence of AM fungi on haustorium formation in a root hemiparasitic plant.

Methods

Using a facultative root hemiparasitic species (Pedicularis tricolor) with the potential to form AM associations, the effects of inoculation were tested with two AM fungal species, Glomus mosseae and Glomus intraradices, on haustorium initiation in P. tricolor grown alone or with Hordeum vulgare ‘Fleet’ (barley) as the host plant. This study consisted of two greenhouse pot experiments.

Key Results

Both AM fungal species dramatically suppressed intraspecific haustorium initiation in P. tricolor at a very low colonization level. The suppression over-rode inductive effects of the parasite''s host plant on haustoria production and caused significant growth depression of P. tricolor.

Conclusions

AM fungi had strong and direct suppressive effects on haustorium formation in the root hemiparasite. The significant role of AM fungi in haustorium initiation of parasitic plants was demonstrated for the first time. This study provides new clues for the regulation of haustorium formation and a route to development of new biocontrol strategies in management of parasitic weeds.     

Organisation of the endosperm and endosperm–placenta syncytia in bladderworts (Utricularia, Lentibulariaceae) with emphasis on the microtubule arrangement

Multinucleate cells play an important role in higher plants, especially during reproduction; however, the configurations of their cytoskeletons, which are formed as a result of mitosis without cytokinesis, have mainly been studied in coenocytes. Previous authors have proposed that in spite of their developmental origin (cell fusion or mitosis without cytokinesis), in multinucleate plant cells, radiating microtubules determine the regular spacing of individual nuclei. However, with the exception of specific syncytia induced by parasitic nematodes, there is no information about the microtubular cytoskeleton in plant heterokaryotic syncytia, i.e. when the nuclei of fused cells come from different cell pools. In this paper, we describe the arrangement of microtubules in the endosperm and special endosperm–placenta syncytia in two Utricularia species. These syncytia arise from different progenitor cells, i.e. cells of the maternal sporophytic nutritive tissue and the micropylar endosperm haustorium (both maternal and paternal genetic material). The development of the endosperm in the two species studied was very similar. We describe microtubule configurations in the three functional endosperm domains: the micropylar syncytium, the endosperm proper and the chalazal haustorium. In contrast to plant syncytia that are induced by parasitic nematodes, the syncytia of Utricularia had an extensive microtubular network. Within each syncytium, two giant nuclei, coming from endosperm cells, were surrounded by a three-dimensional cage of microtubules, which formed a huge cytoplasmic domain. At the periphery of the syncytium, where new protoplasts of the nutritive cells join the syncytium, the microtubules formed a network which surrounded small nuclei from nutritive tissue cells and were also distributed through the cytoplasm. Thus, in the Utricularia syncytium, there were different sized cytoplasmic domains, whose architecture depended on the source and size of the nuclei. The endosperm proper was isolated from maternal (ovule) tissues by a cuticle layer, so the syncytium and chalazal haustorium were the only way for nutrients to be transported from the maternal tissue towards the developing embryo.     

Metabolite changes with induction of Cuscuta haustorium and translocation from host plants

Cuscuta is a stem holoparasitic plant without leaves or roots, parasitizing various types of host plants and causing major problems for certain crops. Cuscuta is known as a generalist and, thus, must have unique parasite strategies to cope with different host plants. For elucidating metabolic responses and mechanisms of parasitization, metabolomic approaches using GC/MS were applied. We compared five stages of Cuscuta japonica: early stage seedlings, with far red light (FR) cue, with contact signal, haustorium induced seedlings by both signals and adult plant parasites on host plants. Sugars, amino acids, organic acids, nucleic acids, and polyols were identified from the polar phase fraction. The apical part contained metabolite profiles different from the haustorium induced part or the basal part. Amino acid and some organic acids were up-regulated for haustorium induction but decreased after parasitization. After attachment to different host plants, metabolite profiles of Cuscuta japonica changed dramatically due to the absorption of specific host plant metabolites such as pinitol. Cuscuta seedlings attached to pinitol rich host plants contained more pinitol and showed different profiles from those attached to plants having less or lacking pinitol.     

Specific Targeting of the Arabidopsis Resistance Protein RPW8.2 to the Interfacial Membrane Encasing the Fungal Haustorium Renders Broad-Spectrum Resistance to Powdery Mildew

Powdery mildew fungal pathogens penetrate the plant cell wall and develop a feeding structure called the haustorium to steal photosynthetate from the host cell. Here, we report that the broad-spectrum mildew resistance protein RPW8.2 from Arabidopsis thaliana is induced and specifically targeted to the extrahaustorial membrane (EHM), an enigmatic interfacial membrane believed to be derived from the host cell plasma membrane. There, RPW8.2 activates a salicylic acid (SA) signaling-dependent defense strategy that concomitantly enhances the encasement of the haustorial complex and onsite accumulation of H₂O₂, presumably for constraining the haustorium while reducing oxidative damage to the host cell. Targeting of RPW8.2 to the EHM, however, is SA independent and requires function of the actin cytoskeleton. Natural mutations that impair either defense activation or EHM targeting of RPW8.2 compromise the efficacy of RPW8.2-mediated resistance. Thus, the interception of haustoria is key for RPW8-mediated broad-spectrum mildew resistance.     

Ultrastructural Studies of the Intercellular Hypha aud Haustorium of Puccinia recondita f.sp. tritici

The ultrastructure of intercellular hyphae and D-hati-storia of P. recondita f.sp. tritici, and the host response to haustorial invasion, was investigated. The intercellular hyphae share common characteristics with those of other uredial stage rust fungi. Anastomosis was observed between intercellular hyphae. Two nucleoli were frequently observed in a single nucleus in the haustorium, indicating possible nuclear fusion between the two nuclei in D-haustoria of this fungus. The close association of host organelles, such as the nucleus, Golgi bodies, endo-plasmic reticulum, vesicles and mitochondria, with the developing haustorium, is described.     

Development of appressoria of Erysiphe graminis f.sp. hordei in response to host plant age and water stress

Appressoria were observed to develop in sequence up to three lobes, each of which may establish a penetration of the underlying host cell and, ultimately, give rise to a haustorium and elongating secondary hyphae (ESH). Upon fully susceptible, well-watered seedling plants increased appressorium development in terms of lobe number led to reduced penetration success, as judged by ESH formation. A concomitant reduction in the initiation of further penetration attempts at lobes newly formed after earlier unsuccessful penetration attempts was also observed. The effects of increased plant age and water stress upon this interaction were examined.