Haustorial Development Research Articles

An interolog-based barley interactome as an integration framework for immune signaling.

The barley MLA nucleotide-binding leucine-rich-repeat (NLR) receptor and its orthologs confer recognition specificity to many fungal diseases, including powdery mildew, stem-, and stripe rust. We used interolog inference to construct a barley protein interactome (Hordeum vulgare predicted interactome, HvInt) comprising 66,133 edges and 7,181 nodes, as a foundation to explore signaling networks associated with MLA.HvInt was compared with the experimentally validated Arabidopsis interactome of 11,253 proteins and 73,960 interactions, verifying that the 2 networks share scale-free properties, including a power-law distribution and small-world network. Then, by successive layering of defense-specific "omics" datasets,HvInt was customized to model cellular response to powdery mildew infection. Integration ofHvInt with expression quantitative trait loci (eQTL) enabled us to infer disease modules and responses associated with fungal penetration and haustorial development. Next, usingHvInt and infection-time-course RNA sequencing of immune signaling mutants, we assembled resistant and susceptible subnetworks. The resulting differentially coexpressed (resistant - susceptible) interactome is essential to barley immunity, facilitates the flow of signaling pathways and is linked to mildew resistance locus a (Mla) through trans eQTL associations. Lastly, we anchoredHvInt with new and previously identified interactors of the MLA coiled coli + nucleotide-binding domains and extended these to additional MLA alleles, orthologs, and NLR outgroups to predict receptor localization and conservation of signaling response. These results link genomic, transcriptomic, and physical interactions during MLA-specified immunity.

Defence responses of native and invasive plants to the native generalist vine parasite Cassytha pubescens – anatomical and functional studies

We investigated the responses of two invasive and two native host species to the parasitic vine Cassytha pubescens R.Br. using glasshouse experiments. We assessed growth of the parasite and its hosts, and anatomy and functionality of haustoria. Target hosts were infected using C. pubescens already established on a donor host. This enabled measurement of growth in target hosts that were detached (parasite connection severed) or not from the donor host. Haustorial connections to hosts were investigated using histological methods. We tested the functionality of haustoria in one invasive and one native host using radiolabelled phosphorus (32P). After it was severed from the donor host, C. pubescens grew poorly on the native host, Acacia myrtifolia (Sm.)Willd. This was likely due to a lack of effective functional haustorial development because although haustoria were firmly attached and morphologically alike those formed on the other hosts, their anatomy was different: their connections with the vascular system were not developed and there was no transfer of 32P from A. myrtifolia to the parasite. In contrast, the other three host species supported the growth of the parasite and had fully developed haustoria. Effective transfer of 32P from the invasive host to the parasite confirmed this. Our results suggest a range of defence mechanisms in C. pubescens hosts and are consistent with reports of strong detrimental effects on invasive hosts. Further, they amount to evidence for the potential use of a native parasite as biological control for invasive species.

Histological and cytological studies of plant infection by Erysiphe euonymi-japonici.

Powdery mildew caused by Erysiphe euonymi-japonici (Eej) is an increasingly serious fungal disease on Euonymus japonicus that is an important ornamental plant. However, little is currently known about infection and pathogenesis of Eej on E. japonicus. Here, we report plant infection by Eej at the histological and cytological levels. Eej caused severe disease symptoms with white and snow-like colonies on leaf surfaces of E. japonicus. Microscopic observations were conducted continuously to define infection process of Eej on E. japonicus. Eej conidia germinated to produce appressorial germ tubes on leaf surfaces and formed irregular haustoria in plant epidermal cells at 6h post-inoculation (hpi) and 12hpi, respectively. After uptaking nutrients from host cells by haustoria, Eej formed numerous hyphae and extensive colonization on leaf surfaces at 96hpi and finally produced abundant conidiophores and new conidia on leaf surfaces at 168hpi. In addition, there was consistently a single nucleus in different Eej infection structures and haustorial development could be divided into three major stages, including formation of penetration peg, formation of haustorial neck and initial haustorium, and maturation of haustorium. These results provide useful information for further determination of Eej pathogenesis and finally controlling the disease.

Interchromosomal Transfer of Immune Regulation During Infection of Barley with the Powdery Mildew Pathogen.

Powdery mildew pathogens colonize over 9500 plant species, causing critical yield loss. The Ascomycete fungus, Blumeria graminis f. sp. hordei (Bgh), causes powdery mildew disease in barley (Hordeum vulgare L.). Successful infection begins with penetration of host epidermal cells, culminating in haustorial feeding structures, facilitating delivery of fungal effectors to the plant and exchange of nutrients from host to pathogen. We used expression Quantitative Trait Locus (eQTL) analysis to dissect the temporal control of immunity-associated gene expression in a doubled haploid barley population challenged with Bgh. Two highly significant regions possessing trans eQTL were identified near the telomeric ends of chromosomes (Chr) 2HL and 1HS. Within these regions reside diverse resistance loci derived from barley landrace H. laevigatum (MlLa) and H. vulgare cv. Algerian (Mla1), which associate with the altered expression of 961 and 3296 genes during fungal penetration of the host and haustorial development, respectively. Regulatory control of transcript levels for 299 of the 961 genes is reprioritized from MlLa on 2HL to Mla1 on 1HS as infection progresses, with 292 of the 299 alternating the allele responsible for higher expression, including Adaptin Protein-2 subunit μ AP2M and Vesicle Associated Membrane Protein VAMP72 subfamily members VAMP721/722. AP2M mediates effector-triggered immunity (ETI) via endocytosis of plasma membrane receptor components. VAMP721/722 and SNAP33 form a Soluble N-ethylmaleimide-sensitive factor Attachment Protein REceptor (SNARE) complex with SYP121 (PEN1), which is engaged in pathogen associated molecular pattern (PAMP)-triggered immunity via exocytosis. We postulate that genes regulated by alternate chromosomal positions are repurposed as part of a conserved immune complex to respond to different pathogen attack scenarios.

Broomrape Weeds. Underground Mechanisms of Parasitism and Associated Strategies for their Control: A Review.

Broomrapes are plant-parasitic weeds which constitute one of the most difficult-to-control of all biotic constraints that affect crops in Mediterranean, central and eastern Europe, and Asia. Due to their physical and metabolic overlap with the crop, their underground parasitism, their achlorophyllous nature, and hardly destructible seed bank, broomrape weeds are usually not controlled by management strategies designed for non-parasitic weeds. Instead, broomrapes are in current state of intensification and spread due to lack of broomrape-specific control programs, unconscious introduction to new areas and may be decline of herbicide use and global warming to a lesser degree. We reviewed relevant facts about the biology and physiology of broomrape weeds and the major feasible control strategies. The points of vulnerability of some underground events, key for their parasitism such as crop-induced germination or haustorial development are reviewed as inhibition targets of the broomrape-crop association. Among the reviewed strategies are those aimed (1) to reduce broomrape seed bank viability, such as fumigation, herbigation, solarization and use of broomrape-specific pathogens; (2) diversion strategies to reduce the broomrape ability to timely detect the host such as those based on promotion of suicidal germination, on introduction of allelochemical interference, or on down-regulating host exudation of germination-inducing factors; (3) strategies to inhibit the capacity of the broomrape seedling to penetrate the crop and connect with the vascular system, such as biotic or abiotic inhibition of broomrape radicle growth and crop resistance to broomrape penetration either natural, genetically engineered or elicited by biotic- or abiotic-resistance-inducing agents; and (4) strategies acting once broomrape seedling has bridged its vascular system with that of the host, aimed to impede or to endure the parasitic sink such as those based on the delivery of herbicides via haustoria, use of resistant or tolerant varieties and implementation of cultural practices improving crop competitiveness.

Construction of a haustorium development associated SSH library in Thesium chinense and analysis of specific ESTs included by Imperata cylindrica

Thesium chinense Turcz. is a facultative hemiparasite that can invade host plants by inducing haustoria on neighboring host roots to access the water and nutrients of the host. Suppression subtractive hybridization (SSH) was performed to identify the differentially expressed genes during haustorial development. A total of 395 recombinants were randomly selected for sequencing. Two hundred seventy-six ESTs were retained after this screening procedure, and after DNA sequencing, 217 ESTs (78.6%) exhibited significant homology with previously identified genes in GenBank. Based on GO annotation, we selected 9 ESTs that may be associated with the development of haustorium and be induced by Imperata cylindrica (L.) Beauv.var.major (Nees)C．E．Hubb. for qRT-PCR. The results revealed that the relative quantities of the 4 ESTs from among the 9 ESTs that involved cell-wall modification (pectin methylesterase, TcPME), signal transduction (auxin-responsive aux/iaa gene, TcAux/IAA) and metabolism (class III peroxidase, TcPrx; phenylalanine ammonia-lyase, TcPAL) all increased simultaneously with the development of the roots of T. chinense and I. cylindrica. These results indicated that the relative quantities of TcPME, TcAux/IAA, TcPrx and TcPAL were affected by the secretions of the I. cylindrica root that T. chinense accreted and that these effects were meaningful to the development of the haustorium of T. chinense.

Growth, photosynthesis and haustorial development of semiparasitic Santalum album L. penetrating into roots of three hosts: a comparative study

Photosynthetic and morphological differences between suitable ( Dalbergia sissoo, Lonicera japonica ) and unsuitable ( Aquilaria sinensis ) hosts in their semiparasitic associations with Santalum album were assessed. Santalum album L. is an important semiparasitic tree whose roots join to host roots, through haustoria, to acquire water and nutrients. Yet little is known how different hosts affect S. album growth, photosynthetic performance and haustorial development. In this study, Dalbergia sissoo Roxb., a leguminous tree, and Lonicera japonica Thunb., a non-leguminous vine, were suitable hosts while Aquilaria sinensis (Lour.) Gilg, a non-leguminous tree, was not. The three host plants were cultivated with sandalwood seedlings for 1 year, inducing significant differences in net photosynthetic rate, stomatal conductance and transpiration rate, plant height, haustorial size and number of haustoria. S. album growth was strongly related to net photosynthetic rate, stomatal conductance and transpiration rate. Well-formed and frequently replaced haustoria in the good host D. sissoo likely enhanced metabolism and nutrient transport. Ultrastructural observations showed differences in the interfacial region and internal structure between finger parenchymal cells and host root cells, vascular tissue, the reaction of host root cells and penetration modes. Among them, the finger parenchymal cells in the haustoria that connected with hosts D. sissoo and L. japonica roots were rich in organelles, including mitochondria, endoplasmic reticulum and lysosomes. Numerous small vacuoles accumulated inside membranes of the host root cells and finger parenchymal cells. Many lysosomes were present at the interfacial region where large-scale digestion of the host root cells in D. sissoo and L. japonica occurred. In contrast, in the finger parenchymal cells of the haustoria that joined to A. sinensis roots, organelles were not that obvious and black-stained material at the interfacial regions was indistinct. The modes of invasion were also different. The finger parenchymal cells penetrated into the roots of hosts D. sissoo and L. japonica by cell membrane fusion, pressure and degrading enzymes while in A. sinensis, host root cells were surrounded by the finger parenchymal cells and were self-digested. Even though mature haustoria could penetrate into the root xylem of hosts D. sissoo and A. sinensis, the haustoria that connected with D. sissoo had a mature xylem while A. sinensis had not yet formed any vessel elements. In all S. album haustoria connected to the three host roots, the vascular system developed some xylem but no phloem was observed.

RNA-Seq analysis identifies key genes associated with haustorial development in the root hemiparasite Santalum album.

Santalum album (sandalwood) is one of the economically important plant species in the Santalaceae for its production of highly valued perfume oils. Sandalwood is also a hemiparasitic tree that obtains some of its water and simple nutrients by tapping into other plants through haustoria which are highly specialized organs in parasitic angiosperms. However, an understanding of the molecular mechanisms involved in haustorium development is limited. In this study, RNA sequencing (RNA-seq) analyses were performed to identify changes in gene expression and metabolic pathways associated with the development of the S. album haustorium. A total of 56,011 non-redundant contigs with a mean contig size of 618 bp were obtained by de novo assembly of the transcriptome of haustoria and non-haustorial seedling roots. A substantial number of the identified differentially expressed genes were involved in cell wall metabolism and protein metabolism, as well as mitochondrial electron transport functions. Phytohormone-mediated regulation might play an important role during haustorial development. Especially, auxin signaling is likely to be essential for haustorial initiation, and genes related to cytokinin and gibberellin biosynthesis and metabolism are involved in haustorial development. Our results suggest that genes encoding nodulin-like proteins may be important for haustorial morphogenesis in S. album. The obtained sequence data will become a rich resource for future research in this interesting species. This information improves our understanding of haustorium development in root hemiparasitic species and will allow further exploration of the detailed molecular mechanisms underlying plant parasitism.

Broadly Conserved Fungal Effector BEC1019 Suppresses Host Cell Death and Enhances Pathogen Virulence in Powdery Mildew of Barley (Hordeum vulgare L.).

The interaction of barley, Hordeum vulgare L., with the powdery mildew fungus Blumeria graminis f. sp. hordei is a well-developed model to investigate resistance and susceptibility to obligate biotrophic pathogens. The 130-Mb Blumeria genome encodes approximately 540 predicted effectors that are hypothesized to suppress or induce host processes to promote colonization. Blumeria effector candidate (BEC)1019, a single-copy gene encoding a putative, secreted metalloprotease, is expressed in haustorial feeding structures, and host-induced gene silencing of BEC1019 restricts haustorial development in compatible interactions. Here, we show that Barley stripe mosaic virus-induced gene silencing of BEC1019 significantly reduces fungal colonization of barley epidermal cells, demonstrating that BEC1019 plays a central role in virulence. In addition, delivery of BEC1019 to the host cytoplasm via Xanthomonas type III secretion suppresses cultivar nonspecific hypersensitive reaction (HR) induced by Xanthomonas oryzae pv. oryzicola, as well as cultivar-specific HR induced by AvrPphB from Pseudomonas syringae pv. phaseolicola. BEC1019 homologs are present in 96 of 241 sequenced fungal genomes, including plant pathogens, human pathogens, and free-living nonpathogens. Comparative analysis revealed variation at several amino acid positions that correlate with fungal lifestyle and several highly conserved, noncorrelated motifs. Site-directed mutagenesis of one of these, ETVIC, compromises the HR-suppressing activity of BEC1019. We postulate that BEC1019 represents an ancient, broadly important fungal protein family, members of which have evolved to function as effectors in plant and animal hosts.

Comparative transcriptome analyses reveal core parasitism genes and suggest gene duplication and repurposing as sources of structural novelty.

The origin of novel traits is recognized as an important process underlying many major evolutionary radiations. We studied the genetic basis for the evolution of haustoria, the novel feeding organs of parasitic flowering plants, using comparative transcriptome sequencing in three species of Orobanchaceae. Around 180 genes are upregulated during haustorial development following host attachment in at least two species, and these are enriched in proteases, cell wall modifying enzymes, and extracellular secretion proteins. Additionally, about 100 shared genes are upregulated in response to haustorium inducing factors prior to host attachment. Collectively, we refer to these newly identified genes as putative “parasitism genes.” Most of these parasitism genes are derived from gene duplications in a common ancestor of Orobanchaceae and Mimulus guttatus, a related nonparasitic plant. Additionally, the signature of relaxed purifying selection and/or adaptive evolution at specific sites was detected in many haustorial genes, and may play an important role in parasite evolution. Comparative analysis of gene expression patterns in parasitic and nonparasitic angiosperms suggests that parasitism genes are derived primarily from root and floral tissues, but with some genes co-opted from other tissues. Gene duplication, often taking place in a nonparasitic ancestor of Orobanchaceae, followed by regulatory neofunctionalization, was an important process in the origin of parasitic haustoria.

The Podosphaera xanthii haustorium, the fungal Trojan horse of cucurbit-powdery mildew interactions

The powdery mildew fungi are obligate biotrophic plant pathogens that develop a specialized structure for parasitism termed haustorium, which is responsible for nutrient uptake and factor exchange with the plant. In this work, we present a detailed microscopy analysis of the haustoria of the cucurbit powdery mildew fungus Podosphaera xanthii, a major limiting factor for cucurbit production worldwide. Despite being located inside plant epidermal cells, transmission electron microscopy (TEM) analysis showed the characteristic highly irregular outline of the extrahaustorial membrane that separates the extrahaustorial matrix of haustoria from the cytoplasm of the plant cell. TEM analysis also revealed the presence of some vesicles and electron-dense plaques of material surrounding the haustoria. In confocal microscopy analysis and aniline blue staining we found a positive correlation between haustorial development and deposition of callose, which is distributed as plaques around haustorial complex. In this study, a method for the isolation of P. xanthii haustoria was also adapted, which permitted the analysis of the formation of haustorial lobes and the visualization of vacuoles and the pool of vesicles inside the haustorial complex. Our findings suggested that the haustorial lobes were responsible for vesicular trafficking and most likely act as the main mediators of the fungus-plant dialogue. All of these findings were integrated into a model of the P. xanthii-host cellular interactions.

The powdery mildew resistance protein RPW8.2 is carried on VAMP721/722 vesicles to the extrahaustorial membrane of haustorial complexes.

Plants employ multiple cell-autonomous defense mechanisms to impede pathogenesis of microbial intruders. Previously we identified an exocytosis defense mechanism in Arabidopsis against pathogenic powdery mildew fungi. This pre-invasive defense mechanism depends on the formation of ternary protein complexes consisting of the plasma membrane-localized PEN1 syntaxin, the adaptor protein SNAP33 and closely sequence-related vesicle-resident VAMP721 or VAMP722 proteins. The Arabidopsis thaliana resistance to powdery mildew 8.2 protein (RPW8.2) confers disease resistance against powdery mildews upon fungal entry into host cells and is specifically targeted to the extrahaustorial membrane (EHM), which envelops the haustorial complex of the fungus. However, the secretory machinery involved in trafficking RPW8.2 to the EHM is unknown. Here we report that RPW8.2 is transiently located on VAMP721/722 vesicles, and later incorporated into the EHM of mature haustoria. Resistance activity of RPW8.2 against the powdery mildew Golovinomyces orontii is greatly diminished in the absence of VAMP721 but only slightly so in the absence of VAMP722. Consistent with this result, trafficking of RPW8.2 to the EHM is delayed in the absence of VAMP721. These findings implicate VAMP721/722 vesicles as key components of the secretory machinery for carrying RPW8.2 to the plant-fungal interface. Quantitative fluorescence recovery after photobleaching suggests that vesicle-mediated trafficking of RPW8.2-yellow fluorescent protein (YFP) to the EHM occurs transiently during early haustorial development and that lateral diffusion of RPW8.2-YFP within the EHM exceeds vesicle-mediated replenishment of RPW8.2-YFP in mature haustoria. Our findings imply the engagement of VAMP721/722 in a bifurcated trafficking pathway for pre-invasive defense at the cell periphery and post-invasive defense at the EHM.

Ontogenesis of the collapsed layer during haustorium development in the root hemi‐parasite Santalum album Linn

The structure and development of collapsed layers of the haustorium were studied in Santalum album Linn. Through light and transmission electron microscopy, it was shown that the collapsed layers originated from starch-containing cells when the haustorium developed an internal gland, thickened gradually and ultimately developed into the mantle, which, combined with the sucker, buckled the host root. We report on the presence of inter-collapsed layers for the first time. These layers develop after penetration into the host and are located between the intrusive tissues and the vascular meristematic region, gradually linking the collapsed layers and remains around the sucker. The proliferation of cells in the meristematic region and the 'host tropism' of cortical layers contribute to pressure within the haustorium and result in development of the collapsed layers. Besides, starch-containing cells that turn into collapsed layers are vulnerable to pressure as they lack a large vacuole, have uneven cell wall thickness and a loose cell arrangement. We proposed that the functions of collapsed layers are to efficiently assure that cell inclusion and energy concentrate at the inner meristematic region and are recycled to affect penetration, reinforce the physical connection between the sandalwood haustorium and host root, and supply space for haustorial development.

Endogenous hormone levels and anatomical characters of haustoria in Santalum album L. seedlings before and after attachment to the host

The physiological and anatomical attributes of haustoria tissues in hemi-parasitic Santalum album L. seedlings, growing on the potential host, Kuhnia rosmarnifolia Vent., were investigated before and after attachment to the host. Quantization of endogenous levels of indole-3-acetic acid (IAA), zeatin (Z), zeatin riboside (ZR), GA-like substances (GAs) and abscisic acid (ABA) was performed by HPLC. Histological preparations were used to characterize structural differences between pre- and post-attachment haustoria. The contents of GAs and ABA were higher in attached haustoria, with 3.61 and 3.50μgg−1 fresh weight, respectively, and three times higher than in non-attached haustoria. Cytokinins, Z, ZR and IAA levels were also high, and their contents in attached haustoria increased 2.04-, 2.17-, and 2.82-fold more, respectively, than in non-attached haustoria. A high auxin-to-cytokinin ratio contributed to haustorial development of S. album. A numerous amount of starch in parenchyma cells around the meristematic region above the haustorial gland and the endophyte tissue of the post-attachment haustoria were reported in a Santalaceae member for the first time. Many lysosomes were present and large-scale digestion of host cells occurred at the interface between the parasite and host. The haustorial penetration in S. album into the host stele was suggested to be a function of mechanical force and enzymatic activity. Analysis of the endogenous hormone levels and the structural characters in S. album haustoria indicated that the haustoria were able to synthesize phytohormones, which appeared to be necessary for cell division and differentiation during haustorial development. These results suggest that endogenous hormones are involved in the haustorial development of S. album and in water and nutrient transport in the host–parasite association.

Parasitic angiosperms, semagenesis and general strategies for plant–plant signaling in the rhizosphere

In addition to their roles in eukaryotic defense and development, reactive oxygen species (ROS) have recently been identified as critical for host attachment by the parasitic angiosperms. In a process known as semagenesis, ROS generated at the root tip of Striga asiatica (L.) Kuntze (Scrophulariaceae) function together with host peroxidases to oxidize monolignols at the host root surface. As a result, para-benzoquinone products accumulate as both necessary and sufficient components for inducing development of the host attachment organ, the haustorium. This event constitutes the critical vegetative/pathogenic transition in the parasite. New evidence is presented that semagenesis occurs broadly in plant-plant signaling. Eudicotyledenous seedlings are more sensitive to the xenognostic benzoquinones than monocots, but general root development, including root elongation, root hair initiation and root hair growth, is impacted in both clades. Specific inhibitors of haustorial development in S. asiatica also inhibit benzoquinone-mediated root development in the non-parasites. These results suggest a common mechanism for benzoquinone perception. Semagenesis enriches our understanding of the mechanisms available for small-molecule underground information exchange among plants. Critical differences in this process, as used by parasitic plants, are beginning to emerge and point towards new strategies for managing parasitic angiosperms in agricultural settings.

Resistance to Striga hermonthica in a maize inbred line derived from Zea diploperennis

Breeding for resistance to Striga in maize (Zea mays), with paucity of donor source and known mechanisms of resistance, has been challenging. Here, post-attachment development of S. hermonthica was monitored on two maize inbreds selected for field resistance and susceptibility reactions to Striga at the International Institute of Tropical Agriculture. Haustorial invasion of the parasite into roots of these inbreds was examined histologically. Morphological differences were observed between roots of the susceptible and the resistant inbreds. The resistant maize had fewer Striga attachments, delayed parasitic development and higher mortality of attached parasites compared with the susceptible inbred. Striga on the susceptible inbred usually penetrated the xylem and showed substantial internal haustorial development. Haustorial ingress on the resistant inbred was often stopped at the endodermis. Parasites able to reach resistant host xylem vessels showed diminished haustorial development relative to those invading susceptible roots. These results suggest that the resistant inbred expresses a developmental barrier and incompatible response against Striga parasitism.

Identification and characterization of sources of resistance to Erysiphe pisi Syd. in Pisum spp.

AbstractA collection of 67 accessions of Pisum species originating from different countries was screened in a glasshouse test for resistance to Erysiphe pisi. All Pisum fulvum accessions were completely resistant. Incomplete resistance was identified in some accessions of P. sativum subsp. sativum var. arvense and P. sativum subsp. elatius and abyssinicum. Microscopy revealed several distinct cellular mechanisms governing resistance. In P. fulvum, it was mainly due to a high frequency of cell death that occurred both as a rapid response to attempted infection and a delayed response that followed colony establishment. Cell death following colony establishment was also key to the incomplete resistance in some accessions of P. sativum subsp. sativum var. arvense. In addition, impaired spore germination, and to a lesser extent appressorium formation, contributed to pre‐penetration resistance in some accessions. In some cases, resistance also retarded colony growth, possibly through effects on haustorial development or function in cells that survived the attack. Thus, these wild pea accessions offer diverse resistances that could be introduced to cultivated peas to increase the efficacy of powdery mildew resistance.

Novel sources of resistance to Striga hermonthica in Tripsacum dactyloides, a wild relative of maize.

• The parasitic weed Striga hermonthica lowers cereal yield in small-holder farms in Africa. Complete resistance in maize to S. hermonthica infection has not been identified. A valuable source of resistance to S. hermonthica may lie in the genetic potential of wild germplasm. • The susceptibility of a wild relative of maize, Tripsacum dactyloides and a Zea mays-T. dactyloides hybrid to S. hermonthica infection was determined. Striga hermonthica development was arrested after attachment to T. dactyloides. Vascular continuity was established between parasite and host but there was poor primary haustorial tissue differentiation on T. dactyloides compared with Z. mays. Partial resistance was inherited in the hybrid. • Striga hermonthica attached to Z. mays was manipulated such that different secondary haustoria could attach to different hosts. Secondary haustoria formation was inhibited on T. dactyloides, moreover, subsequent haustoria formation on Z. mays was also impaired. • Results suggest that T. dactyloides produces a signal that inhibits haustorial development: this signal may be mobile within the parasite haustorial root system.

Differential effects of D -mannose and 2-deoxy- D -glucose on attempted powdery mildew fungal infection of inappropriate and appropriate G ramineae

Barley, oat and wheat were used as both inappropriate hosts (IH) and appropriate hosts (AH) for three formae speciales of the fungus Blumeria graminis, the causal agent of powdery mildew disease. Treatment with either the glucose analog 2-deoxy- D -glucose (DDG) or with D -mannose dramatically suppressed penetration resistance in IH and to a much lesser extent in AH combinations. Other effects of DDG andD -mannose were strikingly dissimilar. DDG greatly reduced localized autofluorescence at fungal attack sites on epidermal cells, and prevented hypersensitive epidermal cell death (HR). D -mannose had little effect on autofluorescence or HR. DDG arrested the development of fungal haustoria and apparently prohibited biotrophy leading to secondary hyphae. D -mannose allowed haustorial development and functional biotrophy leading to the production of elongating secondary hyphae. This suggests that B. graminis is in some way capable of utilizing D -mannose as a carbon substrate. Results with IH combinations paralleled those of known mlo -barley responses to DDG and D -mannose. Results are discussed in relation to specific physiological processes known to be influenced by either DDG or by D -mannose, or by both compounds.

Control of witchweed Striga hermonthica by intercropping with Desmodium spp., and the mechanism defined as allelopathic.

During investigations into the control of insect damage to maize crops in subsistence farming in Kenya, which involved intercropping with repellent plants, the fodder legumes silverleaf (Desmodium uncinatum) and greenleaf (D. intortum) were also found to reduce dramatically the infestation of maize by parasitic witchweeds such as Striga hermonthica. This effect was confirmed by further field testing and shown to be significantly greater than that observed with other legumes, e.g., cowpea, as were the concomitant yield increases. The mechanism was investigated, and although soil shading and addition of nitrogen fertilizer showed some benefits against S. hermonthica infestation, a putative allelopathic mechanism for D. uncinatum was observed. In screenhouse studies, a highly significant reduction in S. hermonthica infestation was obtained when an aqueous solution, eluting from pots in which D. uncinatum plants were growing, was used to irrigate pots of maize planted in soil seeded with high levels of S. hermonthica. Growth of the parasitic weed was almost completely suppressed. whereas extensive infestation occurred with the control eluate. Laboratory investigations into the allelopathic effect of D. uncinatum, using samples of water-soluble chemical components exuded from cleaned roots, demonstrated that this involved a germination stimulant for S. hermonthica and also an inhibitor for haustorial development.