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Mediterranean-type ecosystems are recognized as critical hotspots for both biodiversity and climate change. Within these environments, plants often interact with diverse species, including holoparasitic plants, while simultaneously facing increasing episodes of precipitation shortages and rising temperatures. Here, we investigated the impact of Orobanche latisquama Reut. ex Boiss infestation on the Mediterranean shrub Salvia rosmarinus (L.) Spenn (rosemary) across three populations along an altitudinal gradient, focusing on its effects on host tolerance and resilience to severe summer drought in its natural habitat. Results showed no major physiological impact of the parasite on the host during spring but revealed an enhanced photo- and antioxidant-protective response during the summer drought in rosemary plants infested with O. latisquama. Infested plants showed elevated contents of α-tocopherol and a shift in the ascorbate ratio towards its oxidized state during the summer, particularly in upper and sun-exposed leaves. This was accompanied by elevated malondialdehyde content, indicating enhanced lipid peroxidation. However, despite the heightened photo-oxidative stress observed in leaves from infested plants, no damage to photosystem II was observed, indicating a good tolerance of rosemary to the interaction between parasitism and drought. By autumn, all plants displayed similar recovery patterns, and the differences between infested and non-infested plants disappeared, thus indicating a high resilience to the combination of these biotic and abiotic stresses. Overall, these findings underscore the great adaptive mechanisms S. rosmarinus plants have evolved to endure severe summer drought, even when challenged by holoparasitic plant infestation, and provide new insights into plant-parasite interactions in Mediterranean-type ecosystems., (© 2024 The Author(s). Physiologia Plantarum published by John Wiley & Sons Ltd on behalf of Scandinavian Plant Physiology Society.)

Key Message: This study provided a non-destructive detection method with Vis-NIR hyperspectral imaging combining with physio-biochemical parameters in Helianthus annuus in response to Orobanche cumana infection that took insights into the monitoring of sunflower weed. Sunflower broomrape (Orobanche cumana Wallr.) is an obligate weed that attaches to the host roots of sunflower (Helianthus annuus L.) leading to a significant reduction in yield worldwide. The emergence of O. cumana shoots after its underground life-cycle causes irreversible damage to the crop. In this study, a fast visual, non-invasive and precise method for monitoring changes in spectral characteristics using visible and near-infrared (Vis-NIR) hyperspectral imaging (HSI) was developed. By combining the bands sensitive to antioxidant enzymes (SOD, GR), non-antioxidant enzymes (GSH, GSH + GSSG), MDA, ROS (O 2 - , OH - ), PAL, and PPO activities obtained from the host leaves, we sought to establish an accurate means of assessing these changes and conducted imaging acquisition using hyperspectral cameras from both infested and non-infested sunflower cultivars, followed by physio-biochemical parameters measurement as well as analyzed the expression of defense related genes. Extreme learning machine (ELM) and convolutional neural network (CNN) models using 3-band images were built to classify infected or non-infected plants in three sunflower cultivars, achieving accuracies of 95.83% and 95.83% for the discrimination of infestation as well as 97.92% and 95.83% of varieties, respectively, indicating the potential of multi-spectral imaging systems for early detection of O. cumana in weed management., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Broomrape (Orobanche cumana) negatively affects sunflower, causing severe yield losses, and thus, there is a need to control O. cumana infestation. Brassinosteroids (BRs) play key roles in plant growth and provide resilience to weed infection. This study aims to evaluate the mechanisms by which BRs ameliorate O. cumana infection in sunflower (Helianthus annuus). Seeds were pretreated with BRs (1, 10, and 100 nM) and O. cumana inoculation for 4 weeks under soil conditions. O. cumana infection significantly reduced plant growth traits, photosynthesis, endogenous BRs and regulated the plant defence (POX, GST), BRs signalling (BAK1, BSK1 to BSK4) and synthesis (BRI1, BR6OX2) genes. O. cumana also elevated the levels of malondialdehyde (MDA), hydroxyl radical (OH - ), hydrogen peroxide (H 2 O 2 ) and superoxide (O 2 •- ) in leaves/roots by 77/112, 63/103, 56/97 and 54/89%, as well as caused ultrastructural cellular damages in both leaves and roots. In response, plants activated a few enzymes, superoxide dismutase (SOD), peroxidase (POD) and reduced glutathione but were unable to stimulate the activity of ascorbate peroxidase (APX) and catalase (CAT) enzymes. The addition of BRs (especially at 10 nM) notably recovered the ultrastructural cellular damages, lowered the production of oxidative stress, activated the key enzymatic antioxidants and induced the phenolic and lignin contents. The downregulation in the particular genes by BRs is attributed to the increased resilience of sunflower via a susceptible reaction. In a nutshell, BRs notably enhanced the sunflower resistance to O. cumana infection by escalating the plant immunity responses, inducing systemic acquired resistance, reducing oxidative or cellular damages, and modulating the expression of BR synthesis or signalling genes., (© 2024 Scandinavian Plant Physiology Society.)

Root parasitic weeds of the genera Orobanche and Phelipanche (commonly named broomrapes) are responsible for enormous yield losses of several crops all around the world. Traditional weed management methods, including among others the use of herbicides, soil fumigation and solarization, and mechanical, agronomic or physical methods, may have limits of use or can provide a modicum of control. Difficulties in controlling parasitic weeds are due to both the enormous number of seeds produced by each plant that can remain viable for many years, even in the absence of a host, and to the unique physiological and biological properties of the parasite. Although long considered a suitable and promising approach, biological control, in particular the use of microbial organisms or compounds stimulating or inhibiting seed germination, has had no commercial success and no products have reached the market. This article provides a quick overview of the bioherbicide approaches attempted until now, briefly discussing the causes of the failures and the possibility to improve biocontrol agents' effectiveness. Indeed, despite the failures, the 'bioherbicide' approach deserves renewed interest in light of the enormous scientific and technological progress made in past years, which offers new chances of success. © 2023 The Author. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2023 The Author. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Parasitic weeds such as broomrapes (Phelipanche ramosa and Orobanche cumana) cause severe damage to crops and their development must be controlled. Given that phloroglucinol compounds (PGCs) produced by environmental Pseudomonas could be toxic towards certain plants, we assessed the potential herbicidal effect of the bacterial model Pseudomonas ogarae F113, a PGCs-producing bacterium, on parasitic weed. By combining the use of a mutagenesis approach and of pure PGCs, we evaluated the in vitro effect of PGC-produced by P. ogarae F113 on broomrape germination and assessed the protective activity of a PGC-producing bacteria on oilseed rape (Brassica napus) against P. ramosa in non-sterile soils. We showed that the inhibition of the germination depends on the PGCs molecular structure and their concentrations as well as the broomrape species and pathovars. This inhibition caused by the PGCs is irreversible, causing a brown coloration of the broomrape seeds. The inoculation of PGCs-producing bacteria limited the broomrape infection of P. ramosa, without affecting the host growth. Moreover, elemental profiling analysis of oilseed rape revealed that neither F113 nor applied PGCs affected the nutrition capacity of the oilseed rape host. Our study expands the knowledge on plant-beneficial Pseudomonas as weed biocontrol agents and opens new avenues for the development of natural bioherbicides to enhance crop yield., (© 2023 The Authors. Microbial Biotechnology published by Applied Microbiology International and John Wiley & Sons Ltd.)

Branched broomrape (Phelipanche ramosa (L.) Pomel) is an achlorophyllous root parasitic plant with a wide host range. Its complex management is leading to the abandonment of tobacco or oilseed rape cultivation in the most affected regions in France. Among broomrape regulation factors, soil microorganisms such as fungi seem to be a relevant biocontrol lever. The aim of this work was to detect potential mycoherbicides among fungal endophytic colonizers of P. ramosa parasitizing tobacco. Our hypothesis was that both the inhibitory of broomrape seed germination and the necrotic activities are characteristic of the fungal isolates whatever their taxonomic position. To test this hypothesis, we analysed the taxonomic and functional diversity of fungal isolates of symptomatic P. ramosa collected from infested tobacco-growing regions in France in order to identify one or more fungal strains for future biocontrol. The fungal isolates were characterized using morphological and molecular identification tools and tested for their ability to inhibit the germination of P. ramosa seeds, their necrotic activity on the stems of the pest and their non-pathogenicity to the host plant. We highlighted the specific richness of fungal colonizers associated with symptomatic P. ramosa. Among the 374 collected isolates, nearly 80% belonged to 19 Fusarium species. Eighty-seven isolates representative of this diversity also showed functional diversity by inhibiting seed germination of the parasite. The 20 best-performing isolates showed differences in germination inhibition of P. ramosa at the intraspecific level. Among these 20 fungal isolates, a set of 15 randomly selected isolates was tested for their necrotic activity on the parasite stems. Fusarium venenatum isolates showed dual competence, i.e. germination inhibition and necrotic activity, and were non-pathogenic to tobacco. This led us to discuss the potential mycoherbicidal effect of this fungal species on P. ramosa., (© The Author(s) 2022. Published by Oxford University Press on behalf of FEMS.)

The basic helix-loop-helix (bHLH) transcription factors play important roles in many processes such as plant growth, metabolism and response to biotic/abiotic stresses. Sunflower (Helianthus annuus) is a major oil crop, cultivated throughout the world. However, no systematic characterization of bHLH gene members in sunflower (HabHLH) and their functions involved in drought, cadmium tolerance and Orobanche cumana resistance has been reported yet. In this study, 183 HabHLH genes were identified and named according to their chromosomal locations. We classified these proteins into 21 subfamilies by phylogenetic tree analysis. Subsequently, DNA-binding patterns, sequence analysis, duplication analysis and gene structures were analyzed. All of the HabHLH genes were randomly distributed on 17 chromosomes, and 10 pairs of tandem duplicated genes and one pair of segmental duplicated genes were detected in the HabHLH family. Among the duplicated gene pairs, eight pairs of HabHLH genes suffer from positive selection. Moreover, qRT-PCR results revealed significant up-regulated expression of HabHLH024 gene in response to both abiotic (cadmium, drought) and biotic (Orobanche cumana) stresses, suggesting its important functions in response to different stresses. Therefore, HabHLH024 would be the potential candidate gene for the sunflower tolerance breeding., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Orobanche cumana is a holoparasitic plant that attaches to host-plant roots and seriously reduces the yield of sunflower (Helianthus annuus L.). Effective control methods are lacking with only a few known sources of genetic resistance. In this study, a seed-soak agroinoculation (SSA) method was established, and recombinant tobacco rattle virus vectors were constructed to express RNA interference (RNAi) inducers to cause virus-induced gene silencing (VIGS) in sunflower. A host target gene HaTubulin was systemically silenced in both leaf and root tissues by the SSA-VIGS approach. Trans-species silencing of O. cumana genes were confirmed for 10 out of 11 target genes with silencing efficiency of 23.43%-92.67%. Knockdown of target OcQR1, OcCKX5, and OcWRI1 genes reduced the haustoria number, and silencing of OcEXPA6 caused further phenotypic abnormalities such as shorter tubercles and necrosis. Overexpression of OcEXPA6 caused retarded root growth in alfalfa (Medicago sativa). The results demonstrate that these genes play an important role in the processes of O. cumana parasitism. High-throughput small RNA (sRNA) sequencing and bioinformatics analyses unveiled the distinct features of target gene-derived siRNAs in O. cumana such as siRNA transitivity, strand polarity, hotspot region, and 21/22-nt siRNA predominance, the latter of which was confirmed by Northern blot experiments. The possible RNAi mechanism is also discussed by analyzing RNAi machinery genes in O. cumana. Taken together, we established an efficient host-induced gene silencing technology for both functional genetics studies and potential control of O. cumana. The ease and effectiveness of this strategy could potentially be useful for other species provided they are amenable to SSA., (© American Society of Plant Biologists 2020. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Although it is well known that parasitic weeds such as Orobanche (broomrape) significantly reduce the yield of economically important crops, their infection-induced oxidative changes need more exploration in their host plants. Moreover, applying an eco-friendly approach to minimize the infection is not yet available. This study was conducted to understand the effect of Orobanche ramosa infection on oxidative and redox status of tomato plants and the impact of hormonal (indole acetic acid (IAA); 0.09 mM and salicylic acid (SA); 1.0 mM) seed-priming upon mitigating the infection threats. Although Orobanche invades tomato roots, its inhibitory effects on shoot biomass were also indicted. Orobanche infection usually induces oxidative damage i.e., high lipid peroxidation, lipoxygenase activity and H 2 O 2 levels, particularly for roots. Interestingly, hormonal seed-priming significantly enhanced tomato shoots and roots growth under both healthy and infected conditions. Also, IAA and SA treatment significantly reduced Orobanche infection-induced oxidative damage. The protective effect of seed-priming was explained by increasing the antioxidant defense markers including the antioxidant metabolites (i.e., total antioxidant capacity, carotenoids, phenolics, flavonoids, ASC, GSH, tocopherols) and enzymes (CAT, POX, GPX, SOD, GR, APX, MDHAR, DHAR), particularly in infected tomato seedlings. Additionally, cluster analysis indicated the differential impact of IAA- and SA-seed-priming, whereas lower oxidative damage and higher antioxidant enzymes' activities in tomato root were particularly reported for IAA treatment. The principal component analysis (PCA) also proclaimed an organ specificity depending on their response to Orobanche infection. Collectively, here and for the first time, we shed the light on the potential of seed-priming with either IAA or SA to mitigate the adverse effect of O. ramosa stress in tomato plants, especially at oxidative stress levels., Competing Interests: Declaration of Competing Interest The authors have declared no conflict of interest., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

It is well known that parasitic weeds such as Orobanche (broomrape) significantly decrease crop growth and yield. Although hormonal priming is a well-known inducer of plant resistance against broomrapes (Orobanche spp.), the metabolic events associated with such resistance are poorly understood. Therefore, the current work was undertaken to elucidate the role of SA in inducing tomato resistance against Orobanche, considering its impact on carbon and nitrogen metabolism of the host. Total carbon and nitrogen and levels of carbon (sugars, organic acids and fatty acids) and nitrogen (amino acids and polyamines)-containing metabolites as well as the activities of some key enzymes involved in their metabolic pathways were evaluated. Broomrape infection significantly disrupted C/N ratio in the host roots. On contrary, SA treatment markedly induced accumulation of sugars, organic acids, fatty acids, amino acids as well as polyamines in healthy plants. Under broomrape challenge, SA mitigated the infection-induced growth inhibition by improving the level of nitrogen-containing osmoprotectants (proline, arginine and some polyamines). However, a decrease was observed in some C and N assimilates which are well known to be potentially transferred to the parasite, such as sucrose, asparagine, alanine, serine and glutamate. Interestingly, SA treatment induced the catapolism of polyamines and fatty acids in the host root. Accordingly, our study suggests that SA-induced resistance against broomrape relies on the rational utilization of C and N assimilates in a manner that disturbs the sink strength of the parasite and/or activates the defense pool of the host., Competing Interests: Declaration of competing interest The authors have declared no conflict of interest., (Copyright © 2019. Published by Elsevier Masson SAS.)

Broomrape is a root parasitic plant causing yield losses in sunflower production. Since sunflower is an important oil crop, the development of broomrape-resistant hybrids is the prime breeding objective. Using conventional plant breeding methods, breeders have identified resistant genes and developed a number of hybrids resistant to broomrape, adapted to different growing regions worldwide. However, the spread of broomrape into new countries and the development of new and more virulent races have been noted intensively. Recent advances in sunflower genomics provide additional tools for plant breeders to improve resistance and find durable solutions for broomrape spread and virulence. This review describes the structure and distribution of new, virulent physiological broomrape races, sources of resistance for introduction into susceptible cultivated sunflower, qualitative and quantitative resistance genes along with gene pyramiding and marker assisted selection (MAS) strategies applied in the process of increasing sunflower resistance. In addition, it presents an overview of underutilized biotechnological tools, such as phenotyping, -omics, and genome editing techniques, which need to be introduced in the study of sunflower resistance to broomrape in order to achieve durable resistance.

Six faba bean parents and their F 1 and F 2 generations were used in this investigation to study the genetic system controlling resistance of faba bean (Vicia faba L.) to broomrape (Orobanche crenata). Most of the F 1 hybrids were tolerant to broomrape. In the F 2 generation, the population P 5  × P 6 (Assiut 125 × Romy 12) gave the highest value of relative yield and tolerance index. Heterosis and inbreeding depression were only positive in number of tillers/plant and seed yield/plant characters. The results indicated that the additive effect was more important than the dominance one (D > H 1 ) only for No. of pods/plant in the F 1 generation. Moreover, the narrow-sense heritability was low for most of the studied traits. Three molecular marker systems, namely RAPD, ISSR and SRAP were used for identification and estimation of the genetic diversity among the six faba bean genotypes. The three molecular markers generated DNA unique bands for all genotypes. Only, eight DNA fragments were related to Orobanche tolerance. Clearly and reproducible polymorphic markers were subjected to QTL analysis. The linkage analysis showed that, out of 34 marker loci segregated in the F 2 population, 29 (85.29%) were mapped on three linkage groups. QTL analysis using SIM method performed for the 29 markers assigned to LG-1, LG-2 and LG-3 with the eight traits, number of tillers/plant, plant height, number of pods/plant, seed yield/plant, number of broomrape spikes per plant, height of broomrape spikes, relative yield and tolerance index, showing 12 putative QTLs for all traits except number of tillers/plant. From this study, it is clear that P 5  × P 6 (Assiut 125 × Romy12) population could be considered promising for selection for resistance to broomrape infestation.

Background: Strigolactones (SLs) have a vast number of ecological implications because of the broad spectrum of their biological activities. Unfortunately, the limited availability of SLs restricts their applicability for the benefit of humanity and renders synthesis the only option for their production. However, the structural complexity of SLs impedes their economical synthesis, which is unfeasible on a large scale. Synthesis of SL analogues and mimics with a simpler structure, but with retention of bioactivity, is the solution to this problem., Results: Here, we present eight new hybrid-type SL analogues derived from auxin, synthesized via coupling of auxin ester [ethyl 2-(1H-indol-3-yl)acetate] and of ethyl 2-phenylacetate with four D-rings (mono-, two di- and trimethylated). The new hybrid-type SL analogues were bioassayed to assess the germination activity of seeds of the parasitic weeds Striga hermonthica, Orobanche minor and Phelipanche ramosa using the classical method of counting germinated seeds and a colorimetric method. The bioassays revealed that analogues with a natural monomethylated D-ring had appreciable to good activity towards the three species and were the most active derivatives. By contrast, derivatives with the trimethylated D-ring showed no activity. The dimethylated derivatives (2,4-dimethyl and 3,4-dimethyl) were slightly active, especially towards P. ramosa., Conclusions: New hybrid-type analogues derived from auxins have been prepared. These analogues may be attractive as potential suicidal germination agents for parasitic weed control because of their ease of preparation and relevant bioactivity. © 2019 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2019 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Industrial waste sites, although extremely difficult to revegetate, may be suitable for rare plants such as Orobanche lutea that are condemned to extinction due to their low ability to compete in their natural habitats. The presence of potentially toxic metals seems to facilitate the expansion of O. lutea (parasitizing Medicago falcata) and was found in hundreds of exemplars per m 2 in south Poland and potentially could spread to other localities, causing yield loss in agricultural plants. The main aim of this research was to characterize the interaction between the host, the parasitic plant and symbiotic microbes under different metal concentration in the substratum. The parasite was more common on more polluted soil and when the parasite was connected to the host, potentially toxic metals (Zn, Cd and Pb) were shared by both plants; thus, the content and concentration of these potentially toxic metals in the host were lower than those in plants without parasites. While the performance index (PI ABS ) of photosynthesis was lower in parasitized plants on control soil, on metal-rich industrial waste soil, PI ABS was higher in the parasitized plants than in cases where M. falcata grew alone. This result suggests a role of this parasite in toxicity attenuation, although the biomass of parasitized plants and those growing on polluted sites was lower than that in control sites. In the described case, mycorrhizal colonization and arbuscular richness in M. falcata were even more highly developed on polluted sites than in control ones. The data presented support the hypothesis that the expansion of O. lutea is most likely supported by the increased concentrations of Zn and Cd in areas connected with industrial waste. Although, on industrial wastes the host yield was decreased in the parasite presence, its photosynthetic capacity was even increased., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Broomrapes (Orobanche sp.) are root holoparasitic plants causing severe damage to various families of horticultural crops. A Preliminary survey was carried in tomato fields infested with orobanche in Potohar plateau of Pakistan. Results indicated that each planting bed comprising10-12 tomato plants contained 7-8 parasitic weeds. Overall survey indicated the 65-70% plants suffered partial or complete yield loss. Further, it was estimated that the number of weeds inhabiting plant-1 was ranged from 2-4. Yield loss was estimated at 57.17 tons/ha while in terms of financial loss, it was 2579.87 $/ha. Hence total estimated loss on the total surveyed area of 16 hectares was 41395.85 $. The high potential of tomato cultivation in Pakistan reinforces the urgent protection measures to resolve this emerging issue.

Strigolactones (SLs), a group of plant hormones, induce germination of root-parasitic plants and inhibit shoot branching in many plants. Shoot branching is an important trait that affects the number and quality of flowers and fruits. Root-parasitic plants, such as Phelipanche spp., infect tomato roots and cause economic damage in Europe and North Africa-hence why resistant tomato cultivars are needed. In this study, we found carotenoid cleavage dioxygenase 8- defective mutants of Micro-Tom tomato ( slccd8 ) by the "targeting induced local lesions in genomes" (TILLING) method. The mutants showed excess branching, which was suppressed by exogenously applied SL. Grafting shoot scions of the slccd8 mutants onto wild-type (WT) rootstocks restored normal branching in the scions. The levels of endogenous orobanchol and solanacol in WT were enough detectable, whereas that in the slccd8 mutants were below the detection limit of quantification analysis. Accordingly, root exudates of the slccd8 mutants hardly stimulated seed germination of root parasitic plants. In addition, SL deficiency did not critically affect the fruit traits of Micro-Tom. Using a rhizotron system, we also found that Phelipanche aegyptiaca infection was lower in the slccd8 mutants than in wild-type Micro-Tom because of the low germination. We propose that the slccd8 mutants might be useful as new tomato lines resistant to P. aegyptiaca .

The obligate root parasitic weeds commonly known as broomrape (Orobanche and Phelipanche spp.) cause severe damage to vegetable and field crops worldwide. Efficient control of these parasites is difficult due to their development and attachment to the host plant (via a specialized organ, the haustorium) under the soil surface and to their unique biological traits of massive seed production, facile seed dispersal, germination only under specific conditions, and seed longevity. The major damage inflicted by the parasites takes place underground, making control extremely challenging. Egyptian broomrape (Phelipanche aegyptiaca) is a devastating pest in the Mediterranean basin, parasitizing a wide host crop range, including tomato, sunflower, legumes, and carrot, resulting in severe crop losses. Twenty years of research have led to the development of integrated smart management strategies for combating this parasite in processing tomato fields. In particular, an explicit decision support system (DSS) designated PICKIT has been developed; this DSS is based on predicting parasitism dynamics and employing a range of selective targeted chemical applications (preplanting incorporation, foliar application, and herbigation). In this feature article, we describe the evolution of this research from the laboratory, through greenhouse and experimental field trials, to large scale commercial fields and the successful assimilation of PICKIT into agricultural practice. The use of PICKIT in fields of processing tomatoes in northern Israel has led to effective control of Egyptian broomrape, even in fields with high infestation levels, resulting in a tomato yield increase of an average of 40 tons ha -1 compared with nontreated plots. In 2016, PICKIT was commercially implemented in 33 fields, totaling 400 ha, giving 95% Egyptian broomrape control and tomato yields of 115 to 145 tons ha -1 . The outcome of this research is now enabling farmers to grow tomatoes in Egyptian broomrape-infested fields with assured increased yields and hence high profits.

Purpose: Broomrape produces serious damage to many legume crops and, particularly, becomes a limiting factor for faba bean (Vicia faba L.) production in the Mediterranean basin. Currently, several traditional methods of control have been developed, but none has proved to be effective for this parasite. However, breeding for resistance to this pest remains as one of the most feasible and environment-friendly methods for managing broomrape, but the mechanisms governing the interaction between the parasite and the host are not yet well understood. Therefore, we studied the behavior and molecular and enzymatic changes associated with the resistance to Orobanche foetida in faba bean mutants, which were obtained through radiation mutagenesis., Materials and Methods: Three faba bean genotypes were used in this study, the variety 'Badï', characterized by high productivity in Orobanche-free soils and susceptibility to O. foetida, and two mutant lines P2M3 and P7M3 (derived from radio mutagenesis program), selected for their higher resistance to O. foetida in a field evaluation. The infection progress and the relative changes in the co-culture response, the enzymatic activities changes and the efficiency of the root extract stimulants from the host plant were followed and evaluated in all the genotypes., Results: Experiments showed that low induction of seed germination is a major component of resistance in these lines against O. foetida. This is confirmed by the in vitro experiments with root exudates. The parallel reduction in infection was accompanied by the continuous enhancement of the peroxidase activity, the polyphenol oxidase activity and the phenylalanine ammonia lyase activity in faba bean roots., Conclusion: These data suggest the contribution of these enzymes in faba bean resistance to O. foetida broomrape induced by the use of gamma rays. Management of Orobanche by way of crop selection, based on these enzyme systems is a possible option.

The heterotrophic lifestyle of parasitic plants relies on the development of the haustorium, a specific infectious organ required for attachment to host roots. While haustorium development is initiated upon chemodetection of host-derived molecules in hemiparasitic plants, the induction of haustorium formation remains largely unknown in holoparasitic species such as Phelipanche ramosa. This work demonstrates that the root exudates of the host plant Brassica napus contain allelochemicals displaying haustorium-inducing activity on P. ramosa germinating seeds, which increases the parasite aggressiveness. A de novo assembled transcriptome and microarray approach with P. ramosa during early haustorium formation upon treatment with B. napus root exudates allowed the identification of differentially expressed genes involved in hormone signaling. Bioassays using exogenous cytokinins and the specific cytokinin receptor inhibitor PI-55 showed that cytokinins induced haustorium formation and increased parasite aggressiveness. Root exudates triggered the expression of cytokinin-responsive genes during early haustorium development in germinated seeds, and bio-guided UPLC-ESI(+)-/MS/MS analysis showed that these exudates contain a cytokinin with dihydrozeatin characteristics. These results suggest that cytokinins constitutively exudated from host roots play a major role in haustorium formation and aggressiveness in P. ramosa., (© The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.)

Background: Orobanche crenata represents a major biotic constraint to production of faba bean and lentil in Morocco. While this parasitic plant attacks both of these crops, the extent to which Orobanche biotypes specialise in parasitising specific crops is unknown. To address this question, we studied O. crenata that grew on different hosts and quantified their host specificity to faba bean and lentil. The virulence of O. crenata populations on each host was investigated through field trials, pot and Petri dishes assays. Genetic diversity of the parasite populations was also assessed through molecular analyses., Results: The two legume species showed distinct patterns of specificity. Faba bean was more susceptible to both O. crenata populations, while the specificity for lentil by lentil-grown O. crenata was evident at the final stage of the parasite life cycle as shown by correspondence factorial analyses. Considerable internal variation (81%) within O. crenata populations parasitising both legume species was observed by molecular analyses, but significant divergence (19%; Ø = 0.189; P = 0.010) among the populations was detected., Conclusion: These results indicate that O. crenata can adapt to specific host species, which is important knowledge when developing integrated pest management practices for parasitic weed control. © 2017 Society of Chemical Industry., (© 2017 Society of Chemical Industry.)

Orobanche cumana is an obligate root parasite causing severe damage to many economically important crops, including sunflowers worldwide. For efficient control measures, it is necessary to understand the resistant mechanism during interaction at molecular level. The present study emphasizes on comparative proteomics to investigate the mechanistic basis of compatible and incompatible interaction of O. cumana with resistant (JY207) and susceptible (TK0409) sunflowers. More than 3500 proteins were identified from two cultivars by iTRAQ analysis. Identified proteins associated with general functions, posttranslational modification, energy production and conversion, carbohydrate transport and metabolism, and signal transduction mechanisms were the most represented category of induced proteins in both cultivars. The resistant interaction was characterized by alteration of defense-related proteins involved in recognition of parasites, accumulation of pathogenesis-related proteins, biosynthesis of lignin, and detoxification of toxic metabolites in JY207 after inoculation. The susceptible interaction was characterized by decreased abundance of proteins involved in biosynthesis and signaling of plant growth regulators including auxin, gibberellin, brassinosteroid, and ethylene in TK0409 after inoculation. The present study provides comprehensive details of proteins and differential modulation of pathways regulated under compatible and incompatible interaction, allowing the identification of important molecular components for development of sustainable resistance against this parasite., (© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background: Some radiation-mutagenised chickpea mutants potentially resistant to the broomrape, Orobanche foetida Poir., were selected through field trials. The objectives of this work were to confirm resistance under artificial infestation, in pots and mini-rhizotron systems, and to determine the developmental stages of broomrape affected by resistance and the relevant resistance mechanisms induced by radiation mutagenesis., Results: Among 30 mutants tested for resistance to O. foetida, five shared strong resistance in both pot experiments and mini-rhizotron systems. Resistance was not complete, but the few individuals that escaped resistance displayed high disorders of shoot development. Results demonstrated a 2-3-fold decrease in stimulatory activity of root exudates towards broomrape seed germination in resistant mutants in comparison with non-irradiated control plants and susceptible mutants. Resistance was associated with an induction of broomrape necrosis early during infection. When infested, most of the resistant mutants shared enhanced levels of soluble phenolic contents, phenylalanine ammonia lyase activity, guaiacol peroxidase activity and polyphenol oxidase activity, in addition to glutathione and notably ascorbate peroxidase gene expression in roots., Conclusion: Results confirmed enhanced resistance in chickpea radiation-mutagenised mutants, and demonstrated that resistance is based on alteration of root exudation, presumed cell-wall reinforcement and change in root oxidative status in response to infection. © 2016 Society of Chemical Industry., (© 2016 Society of Chemical Industry.)

Background and Aims: The broomrapes, Orobanche sensu lato (Orobanchaceae), are common root parasites found across Eurasia, Africa and the Americas. All species native to the western hemisphere, recognized as Orobanche sections Gymnocaulis and Nothaphyllon, form a clade that has a centre of diversity in western North America, but also includes four disjunct species in central and southern South America. The wide ecological distribution coupled with moderate taxonomic diversity make this clade a valuable model system for studying the role, if any, of host-switching in driving the diversification of plant parasites., Methods: Two spacer regions of ribosomal nuclear DNA (ITS + ETS), three plastid regions and one low-copy nuclear gene were sampled from 163 exemplars of Orobanche from across the native geographic range in order to infer a detailed phylogeny. Together with comprehensive data on the parasites' native host ranges, associations between phylogenetic lineages and host specificity are tested., Key Results: Within the two currently recognized species of O. sect. Gymnocaulis, seven strongly supported clades were found. While commonly sympatric, members of these clades each had unique host associations. Strong support for cryptic host-specific diversity was also found in sect. Nothaphyllon, while other taxonomic species were well supported. We also find strong evidence for multiple amphitropical dispersals from central North America into South America., Conclusions: Host-switching is an important driver of diversification in western hemisphere broomrapes, where host specificity has been grossly underestimated. More broadly, host specificity and host-switching probably play fundamental roles in the speciation of parasitic plants., (© The Author 2016. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Crenate broomrape (Orobanche crenata Forsk.) is a devastating parasitic weed threatening the cultivation of legumes around the Mediterranean and in the Middle East. So far, only moderate levels of resistance were reported to occur in pea (Pisum sativum L.) natural germplasm, and most commercial cultivars are prone to severe infestation. Here, we describe the selection of a pea line highly resistant to O. crenata, following the screening of local genetic resources. Time series observations show that delayed emergence of the parasite is an important parameter associated with broomrape resistance. High performance liquid chromatography connected to tandem mass spectrometry analysis and in vitro broomrape germination bioassays suggest that the resistance mechanism might involve the reduced secretion of strigolactones, plant hormones exuded by roots and acting as signaling molecules for the germination of parasitic weeds. Two years of replicated trials in noninfested fields indicate that the resistance is devoid of pleiotropic effects on yield, in contrast to pea experimental mutants impaired in strigolactone biosynthesis and, thus, is suitable for use in breeding programs.

Previous studies in Arabidopsis reported that the MAX2 (more axillary growth 2) gene is a component of the strigolactone (SL) signaling pathway, which regulates a wide range of biological processes, from plant growth and development to environmental stress responses. Orobanche aegyptiaca is a harmful parasitic plant for many economically important crops. Seed germination of O. aegyptiaca is very sensitive to SLs, suggesting that O. aegyptiaca may contain components of the SL signaling pathway. To investigate this hypothesis, we identified and cloned a MAX2 ortholog from O. aegyptiaca for complementation analyses using the Arabidopsis Atmax2 mutant. The so-called OaMAX2 gene could rescue phenotypes of the Atmax2 mutant in various tested developmental aspects, including seed germination, shoot branching, leaf senescence and growth and development of hypocotyl, root hair, primary root and lateral root. More importantly, OaMAX2 could enhance the drought tolerance of Atmax2 mutant, suggesting its ability to restore the drought-tolerant phenotype of mutant plants defected in AtMAX2 function. Thus, this study provides genetic evidence that the functions of the MAX2 orthologs, and perhaps the MAX2 signaling pathways, are conserved in parasitic and non-parasitic plants. Furthermore, the results of our study enable us to develop a strategy to fight against parasitic plants by suppressing the MAX signaling, which ultimately leads to enhanced productivity of crop plants., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Seed dormancy release of the obligate root parasitic plant, Phelipanche ramosa, requires a minimum 4-day conditioning period followed by stimulation by host-derived germination stimulants, such as strigolactones. Germination is then mediated by germination stimulant-dependent activation of PrCYP707A1, an abscisic acid catabolic gene. The molecular mechanisms occurring during the conditioning period that silence PrCYP707A1 expression and regulate germination stimulant response are almost unknown. Here, global DNA methylation quantification associated with pharmacological approaches and cytosine methylation analysis of the PrCYP707A1 promoter were used to investigate the modulation and possible role of DNA methylation during the conditioning period and in the PrCYP707A1 response to GR24, a synthetic strigolactone analogue. Active global DNA demethylation occurs during the conditioning period and is required for PrCYP707A1 activation by GR24 and for subsequent seed germination. Treatment with 5-azacytidine, a DNA-hypomethylating molecule, reduces the length of the conditioning period. Conversely, hydroxyurea, a hypermethylating agent, inhibits PrCYP707A1 expression and seed germination. Methylated DNA immunoprecipitation followed by PCR experiments and bisulfite sequencing revealed that DNA demethylation particularly impacts a 78-nucleotide sequence in the PrCYP707A1 promoter. The results here demonstrate that the DNA methylation status during the conditioning period plays a crucial role independently of abscisic acid in the regulation of P. ramosa seed germination by controlling the strigolactone-dependent expression of PrCYP707A1., (© The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology.)

Strigolactones, first discovered as germination stimulants for parasitic weeds [1], are carotenoid-derived phytohormones that play major roles in inhibiting lateral bud outgrowth and promoting plant-mycorrhizal symbiosis [2-4]. Furthermore, strigolactones are involved in the regulation of lateral and adventitious root development, root cell division [5, 6], secondary growth [7], and leaf senescence [8]. Recently, we discovered the strigolactone transporter Petunia axillaris PLEIOTROPIC DRUG RESISTANCE 1 (PaPDR1), which is required for efficient mycorrhizal colonization and inhibition of lateral bud outgrowth [9]. However, how strigolactones are transported through the plant remained unknown. Here we show that PaPDR1 exhibits a cell-type-specific asymmetric localization in different root tissues. In root tips, PaPDR1 is co-expressed with the strigolactone biosynthetic gene DAD1 (CCD8), and it is localized at the apical membrane of root hypodermal cells, presumably mediating the shootward transport of strigolactone. Above the root tip, in the hypodermal passage cells that form gates for the entry of mycorrhizal fungi, PaPDR1 is present in the outer-lateral membrane, compatible with its postulated function as strigolactone exporter from root to soil. Transport studies are in line with our localization studies since (1) a papdr1 mutant displays impaired transport of strigolactones out of the root tip to the shoot as well as into the rhizosphere and (2) DAD1 expression and PIN1/PIN2 levels change in plants deregulated for PDR1 expression, suggestive of variations in endogenous strigolactone contents. In conclusion, our results indicate that the polar localizations of PaPDR1 mediate directional shootward strigolactone transport as well as localized exudation into the soil., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Faba bean yield is severely constrained in the Mediterranean region and Middle East by the parasitic weeds Orobanche crenata, O. foetida, and Phelipanche aegyptiaca. Seed germination of these weeds is triggered upon recognition of host root exudates. Only recently faba bean accessions have been identified with resistance based in low induction of parasitic seed germination, but the underlying mechanism was not identified. Strigolactones are a group of terpenoid lactones involved in the host recognition by parasitic plants. Our LC-MS/MS analysis of root exudates of the susceptible accession Prothabon detected orobanchol, orobanchyl acetate, and a novel germination stimulant. A time course analysis indicated that their concentration increased with plant age. However, low or undetectable amounts of these germination stimulants were detected in root exudates of the resistant lines Quijote and Navio at all plant ages. A time course analysis of seed germination induced by root exudates of each faba bean accession indicated important differences in the ability to stimulate parasitic germination. Results presented here show that resistance to parasitic weeds based on low strigolactone exudation does exist within faba bean germplasm. Therefore, selection for this trait is feasible in a breeding program. The remarkable fact that low induction of germination is similarly operative against O. crenata, O. foetida, and P. aegyptiaca reinforces the value of this resistance.

Cucumber Mosaic Virus (CMV) is a highly infectious cucumovirus, which infects more than 800 plant species and causes major diseases in greenhouse and field crops worldwide. Parasitic weeds such as Phelipanche aegyptiaca are a major constraint to the production of many crops in the world and the parasite's lifestyle makes control extremely difficult. The parasite seeds can germinate after conditioning and perceiving strigolactones secreted by the host roots. Strigolactones are rhizosphere signaling molecules in plants that are biosynthesized through carotenoid cleavage. In the present study we investigated the possibility of reducing β-carotene and then strigolactone production in the host roots by blocking carotenoid biosynthesis using CMV-infected tobacco. It was found that CMV downregulated the enzyme phytoene desaturase(PDS) and reduced significantly both carotenoid production and Phelipanche infection in tobacco host roots infected with both CMV and P. aegyptiaca. Based on our results (decrease of β-carotene and repression of PDS transcripts in tobacco roots), we hypothesized that the reduction of Phelipanche tubercles and shoots occurred due to an effect of CMV on secondary metabolite stimulators such as strigolacetones. Our study indicated that mass production of the host roots was not affected by CMV; however, most inflorescences of Phelipanche grown on CMV-infected tobacco developed abnormally (deformed shoots and short nodes). Carotenoid biosynthesis inhibitors such as CMV can be used to reduce the production of strigolactones, which will lead to decreased Phelipanche attachment. Interestingly, attenuated CMV strains may provide a safe means for enhancing crop resistance against parasitic weeds in a future plan.

Background and Aims: A fundamental element in the evolution of obligate root-parasitic angiosperms is their ability to germinate only in response to chemical stimulation by roots, to ensure contact with a nearby nourishing host. The aim of this study was to explore inheritance of the unique germination control in this group of plants., Methods: Analysis was made of the segregation of spontaneous (non-induced) germination that appeared in hybrid progenies derived from crosses between Orobanche cernua and O. cumana, which, like all other Orobanche species, are totally dependent on chemical stimulation for the onset of germination, and show negligible spontaneous germination in their natural seed populations., Key Results and Conclusions: F(1) and F(2) seeds did not germinate in the absence of chemical stimulation, but significant spontaneous germination was found in some F(3) seed families. This indicates that the prevention of non-induced germination in Orobanche seeds, i.e. dependence on an external chemical stimulation for seed germination, is genetically controlled, that this genetic control is expressed in a seed tissue with maternal origin (presumably the perisperm that originates from the nucellus) and that genetic variation for this trait exists in Orobanche species. Similar segregation results were obtained in reciprocal crosses, suggesting that stimulated germination is controlled by nuclear genes.

The germination of the obligate root parasites of the Orobanchaceae depends on the perception of chemical stimuli from host roots. Several compounds, collectively termed strigolactones, stimulate the germination of the various Orobanche species, but do not significantly elicit germination of Orobanche cumana, a specific parasite of sunflower. Phosphate starvation markedly decreased the stimulatory activity of sunflower root exudates toward O. cumana, and fluridone - an inhibitor of the carotenoid biosynthesis pathway - did not inhibit the production of the germination stimulant in both shoots and roots of young sunflower plants, indicating that the stimulant is not a strigolactone. We identified the natural germination stimulant from sunflower root exudates by bioassay-driven purification. Its chemical structure was elucidated as the guaianolide sesquiterpene lactone dehydrocostus lactone (DCL). Low DCL concentrations effectively stimulate the germination of O. cumana seeds but not of Phelipanche aegyptiaca (syn. Orobanche aegyptiaca). DCL and other sesquiterpene lactones were found in various plant organs, but were previously not known to be exuded to the rhizosphere where they can interact with other organisms., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The parasitic flowering plants of the genera Orobanche and Phelipanche (broomrape species) are obligatory chlorophyll-lacking root-parasitic weeds that infect dicotyledonous plants and cause heavy economic losses in a wide variety of plant species in warm-temperate and subtropical regions. One of the most effective strategies for broomrape control is crop breeding for broomrape resistance. Previous efforts to find natural broomrape-resistant tomato (Solanum lycopersicon) genotypes were unsuccessful, and no broomrape resistance was found in any wild tomato species. Recently, however, the fast-neutron-mutagenized tomato mutant SL-ORT1 was found to be highly resistant to various Phelipanche and Orobanche spp. Nevertheless, SL-ORT1 plants were parasitized by Phelipanche aegyptiaca if grown in pots together with the susceptible tomato cv. M-82. In the present study, no toxic activity or inhibition of Phelipanche seed germination could be detected in the SL-ORT1 root extracts. SL-ORT1 roots did not induce Phelipanche seed germination in pots but they were parasitized, at the same level as M-82, after application of the synthetic germination stimulant GR24 to the rhizosphere. Whereas liquid chromatography coupled to tandem mass spectrometry analysis of root exudates of M-82 revealed the presence of the strigolactones orobanchol, solanacol, and didehydro-orobanchol isomer, these compounds were not found in the exudates of SL-ORT1. It can be concluded that SL-ORT1 resistance results from its inability to produce and secrete natural germination stimulants to the rhizosphere.

Background: Parasitic angiosperm Orobanche crenata infection represents a major constraint for the cultivation of legumes worldwide. The level of protection achieved to date is either incomplete or ephemeral. Hence, an efficient control of the parasite requires a better understanding of its interaction and associated resistance mechanisms at molecular levels., Results: In order to study the plant response to this parasitic plant and the molecular basis of the resistance we have used a proteomic approach. The root proteome of two accessions of the model legume Medicago truncatula displaying differences in their resistance phenotype, in control as well as in inoculated plants, over two time points (21 and 25 days post infection), has been compared. We report quantitative as well as qualitative differences in the 2-DE maps between early- (SA 27774) and late-resistant (SA 4087) genotypes after Coomassie and silver-staining: 69 differential spots were observed between non-inoculated genotypes, and 42 and 25 spots for SA 4087 and SA 27774 non-inoculated and inoculated plants, respectively. In all, 49 differential spots were identified by peptide mass fingerprinting (PMF) following MALDI-TOF/TOF mass spectrometry. Many of the proteins showing significant differences between genotypes and after parasitic infection belong to the functional category of defense and stress-related proteins. A number of spots correspond to proteins with the same function, and might represent members of a multigenic family or post-transcriptional forms of the same protein., Conclusion: The results obtained suggest the existence of a generic defense mechanism operating during the early stages of infection and differing in both genotypes. The faster response to the infection observed in the SA 27774 genotype might be due to the action of proteins targeted against key elements needed for the parasite's successful infection, such as protease inhibitors. Our data are discussed and compared with those previously obtained with pea 1 and transcriptomic analysis of other plant-pathogen and plant-parasitic plant systems.

The field of nanotechnology opens up novel potential applications for agriculture. Nanotechnology applications are already being explored and used in medicine and pharmacology, but interest for use in crop protection is just starting. The development of nanodevices as smart delivery systems to target specific sites and nanocarriers for controlled chemical release is discussed. Some nanotechnologies can improve existing crop management techniques in the short to medium term. Nanocapsules would help to avoid phytotoxicity on the crop by using systemic herbicides against parasitic weeds. Nanoencapsulation can also improve herbicide application, providing better penetration through cuticles and tissues, and allowing slow and constant release of the active substances. On the other hand, new crop management tools could be developed on the basis of medical applications. Nanoparticles have a great potential as 'magic bullets', loaded with herbicides, chemicals or nucleic acids, and targeting specific plant tissues or areas to release their charge. Viral capsids can be altered by mutagenesis to achieve different configurations and deliver specific nucleic acids, enzymes or antimicrobial peptides acting against the parasites. Many issues are still to be addressed, such as increasing the scale of production processes and lowering costs, as well as toxicological issues, but the foundations of a new plant treatment concept have been laid, and applications in the field of parasitic plant control can be started.

It is necessary to control root parasitic weeds before or as they attach to the crop. This can only be easily achieved chemically with herbicides that are systemic, or with herbicides that are active in soil. Long-term control can only be attained if the crops do not metabolise the herbicide, i.e. have target-site resistance. Such target-site resistances have allowed foliar applications of herbicides inhibiting enol-pyruvylshikimate phosphate synthase (EPSPS) (glyphosate), acetolactate synthase (ALS) (e.g. chlorsulfuron, imazapyr) and dihydropteroate synthase (asulam) for Orobanche control in experimental conditions with various crops. Large-scale use of imazapyr as a seed dressing of imidazolinone-resistant maize has been commercialised for Striga control. Crops with two target-site resistances will be more resilient to the evolution of resistance in the parasite, if well managed.

Strigolactones released from plant roots induce seed germination of root parasitic weeds, witchweeds (Striga spp.) and broomrapes (Orobanche spp.), and hyphal branching of symbiotic arbuscular mycorrhizal (AM) fungi. In addition to these functions in the rhizosphere, strigolactones have recently been shown to be a novel class of plant hormones regulating shoot outgrowth. The natural strigolactones identified so far have the common C-D ring moiety, which is thought to be the essential structure for exhibiting biological activity. The introduction of substitutions on the A-B ring moiety of 5-deoxystrigol, the basic strigolactone, affords various strigolactones, e.g. hydroxylation on C-4, C-5 and C-9 leads to orobanchol, strigol and sorgomol respectively. Then, acetylation and probably other derivatisations of these hydroxy-strigolactones would occur. Although the C-2'-(R) stereochemistry was thought to be an important structural feature for potent germination stimulation activity, 2'-epi-strigolactones were found in root exudates of tobacco, rice, pea and other plant species, indicating that at least some plants produce both epimers.

Background: Pea cultivation is strongly hampered in Mediterranean and Middle East farming systems by the occurrence of Orobanche crenata Forsk. Strategies of control have been developed, but only marginal successes have been achieved. Most control methods are either unfeasible, uneconomical, hard to achieve or result in incomplete protection. The integration of several control measures is the most desirable strategy., Results: [corrected] Recent developments in control are presented and re-evaluated in light of recent developments in crop breeding and molecular genetics. These developments are placed within a framework that is compatible with current agronomic practices., Conclusion: The current focus in applied breeding is leveraging biotechnological tools to develop more and better markers to speed up the delivery of improved cultivars to the farmer. To date, however, progress in marker development and delivery of useful markers has been slow. The application of knowledge gained from basic genomic research and genetic engineering will contribute to more rapid pea improvement for resistance against O. crenata and/or the herbicide.

The structures of naturally occurring germination stimulants for seeds of the parasitic weeds Striga spp. and Orobanche spp. are described. The bioactiphore in this strigolactone family of stimulants is deduced from a structure-activity relationship and shown to reside in the CD part of the stimulant molecule. A molecular mechanism for the initial stages of seed germination is proposed. The influence of stereochemistry on the stimulant activity is significant. Combining this molecular information leads to a model for the design of synthetic strigolactones. Nijmegen-1 is a typical example of a highly active, newly designed synthetic stimulant. The occurrence of natural stimulants not belonging to the strigolactone family, such as cotylenin and parthenolide, is briefly described. The biosynthesis of natural strigolactones from beta-carotene is analysed in terms of isolated and predicted stimulants. This scheme will be helpful in the search for new strigolactones from root exudates. Protein fishing experiments to isolate and characterise the receptor protein using biotin-labelled GR 24 are described. A receptor protein of 60 kD was identified by this method. Nijmegen-1 has been tested as a suicidal germination agent in field trials on tobacco infested by Orobanche ramosa L. The preliminary results are highly rewarding. Finally, some future challenges in synthesis are described. These include synthesising new natural and synthetic stimulants and establishing the molecular connection between strigolactones as germination stimulants, as the branching factor for arbuscular mycorrhizal fungi and as an inhibitor of shoot branching.

Species of Orobanche and Striga are among the most damaging parasitic weed species worldwide, but there are few reliable statistics on the full extent of the economic losses they cause. The distribution, host range and economic importance of the major species of Orobanche and Striga are briefly summarised. A review of literature over the period since 1991 suggests that many million hectares are infested and that the losses amount to $ US billions annually. Unfortunately there are almost no fully reliable figures on which to base these figures precisely. Meanwhile, there is little evidence of any significant change in intensity, range or losses caused over this period. Any reduction in the importance of these damaging weeds is sporadic, and alleviation of the problems is mostly localised. Furthermore, while the importance of Orobanche species may be broadly static, Striga species on cereals continue to become more serious in many countries owing to continued loss of soil fertility. It is suggested that new techniques may be needed for measurement of the extent of losses caused by these genera and their economic impact. There is continued urgency to develop control measures appropriate to the farming systems involved, and to reduce the risk of spread of both groups of parasite to new areas.

Background: Parasitic Orobanchaceae germinate only after receiving a chemical stimulus from roots of potential host plants. A preparatory phase of several days that follows seed imbibition, termed conditioning, is known to be required; thereafter the seeds can respond to germination stimulants. The aim of this study was to examine whether conditioning is essential for stimulant receptivity., Results: Non-conditioned seeds of both Orobanche cumana Wallr. and O. aegyptiaca Pers. [syn. Phelipanche aegyptiaca (Pers.) Pomel] were able to germinate in response to chemical stimulation by GR24 even without prior conditioning. Stimulated seeds reached maximal germination rates about 2 weeks after the onset of imbibition, no matter whether the seeds had or had not been conditioned before stimulation. Whereas the lag time between stimulation and germination response of non-conditioned seeds was longer than for conditioned seeds, the total time between imbibition and germination was shorter for the non-conditioned seeds. Unlike the above two species, O. crenata Forsk. was found to require conditioning prior to stimulation., Conclusions: Seeds of O. cumana and O. aegyptiaca are already receptive before conditioning. Thus, conditioning is not involved in stimulant receptivity. A hypothesis is put forward, suggesting that conditioning includes (a) a parasite-specific early phase that allows the imbibed seeds to overcome the stress caused by failing to receive an immediate germination stimulus, and (b) a non-specific later phase that is identical to the pregermination phase between seed imbibition and actual germination that is typical for all higher plants.

Witchweeds (Striga spp.) and broomrapes (Orobanche spp.) are obligate root parasitic plants on economically important field and horticultural crops. The parasites' seeds are induced to germinate by root-derived chemical signals. The radicular end is transformed into a haustorium which attaches, penetrates the host root and establishes connection with the vascular system of the host. Reactions of Lotus japonicus, a model legume for functional genomics, were studied for furthering the understanding of host-parasite interactions. Lotus japonicus was compatible with Orobanche aegyptiaca, but not with Orobanche minor, Striga hermonthica and Striga gesnerioides. Orobanche minor successfully penetrated Lotus japonicus roots, but failed to establish connections with the vascular system. Haustoria in Striga hermonthica attached to the roots, but penetration and subsequent growth of the endophyte in the cortex were restricted. Striga gesnerioides did not parasitize Lotus japonicus. Among seven mutants of Lotus japonicus (castor-5, har1-5, alb1-1, ccamk-3, nup85-3, nfr1-3 and nsp2-1) with altered characteristics in relation to rhizobial nodulation and mycorrhizal colonization, castor-5 and har1-5 were parasitized by Orobanche aegyptiaca with higher frequency than the wild type. In contrast, Orobanche aegyptiaca tubercle development was delayed on the mutants nup85-3, nfr1-3 and nsp2-1. These results suggest that nodulation, mycorrhizal colonization and infection by root parasitic plants in Lotus japonicus may be modulated by similar mechanisms and that Lotus japonicus is a potential model legume for studying plant-plant parasitism.

Lotus japonicus genes responsive to parasitism by the compatible species Orobanche aegyptiaca and the incompatible species Striga hermonthica were isolated by using the suppression subtractive hybridization (SSH) strategy. O. aegyptiaca and S. hermonthica parasitism specifically induced the expression of genes involved in jasmonic acid (JA) biosynthesis and phytoalexin biosynthesis, respectively. Nodulation-related genes were almost exclusively found among the Orobanche-induced genes. Temporal gene expression analyses revealed that 19 out of the 48 Orobanche-induced genes and 5 out of the 48 Striga-induced genes were up-regulated at 1 dai. Four genes, including putative trypsin protease inhibitor genes, exhibited systemic up-regulation in the host plant parasitized by O. aegyptiaca. On the other hand, S. hermonthica attachment did not induce systemic gene expression.

Background: Vascular continuity is established between a host plant and the root parasite broomrape. It is generally accepted that the direction of vascular continuity results from polar flow of auxin. Our hypothesis was that chemical disruptions of auxin transport and activity could influence the infection of the host by the parasite., Methods: A sterile system for the routine infection of Arabidopsis thaliana seedlings in Nunc cell culture plates by germinated seeds of Orobanche aegyptiaca was developed. This method permitted a quantitative assay of the rate of host infection. The three-dimensional structure of the vascular contacts was followed in cleared tissue. IAA (indole acetic acid) or substances that influence its activity and transport were applied locally to the host root., Results: The orientation of the xylem contacts showed that broomrape grafts itself upon the host by acting hormonally as a root rather than a shoot. Local applications of IAA, PCIB (p-chlorophenoxyisobutyric acid) or NPA (naphthylphthalamic acid) all resulted in drastic reductions of Orobanche infection, Conclusions: Broomrape manipulates the host by acting as a sink for auxin. Disruption of auxin action or auxin flow at the contact site could be a novel basis for controlling infection by Orobanche.

Orobanche crenata is a major threat to grain legume production. Fenugreek (Trigonella foenum-graecum) is an annual legume that has been shown to effectively reduce O. crenata infection when intercropped with grain legumes. In this paper, we point that this can be attributed to allelopathy, through inhibition of the germination of O. crenata by fenugreek root exudates. The main inhibitory metabolite was isolated and characterized. Allelopathy was demonstrated in different bioassays, by inhibition of O. crenata seeds germination both by growing fenugreek and pea plants together (intercropped), and by application of fenugreek root exudates. Fenugreek root exudates were extracted with organic solvent and fractionated giving several fractions, two of which showed moderate (27%) and strong (54%) inhibition of O. crenata seed germination, respectively. The most active metabolite is a new monosubstituted trioxazonane, characterized by spectroscopic methods as the 2-butyl-[1,4,7,2]trioxazonane and named trigoxazonane.

Crenate broomrape (Orobanche crenata) is a root parasitic weed that represents a major constraint for grain legume production in Mediterranean and West Asian countries. Medicago truncatula has emerged as an important model plant species for structural and functional genomics. The close phylogenic relationship of M. truncatula with crop legumes increases its value as a resource for understanding resistance against Orobanche spp. Different cytological methods were used to study the mechanisms of resistance against crenate broomrape of two accessions of M. truncatula, showing early and late acting resistance. In the early resistance accession (SA27774) we found that the parasite died before a tubercle had formed. In the late resistance accession (SA4327) the parasite became attached without apparent problems to the host roots but most of the established tubercles turned dark and died before emergence. The results suggest that there are defensive mechanisms acting in both accessions but with a time gap that is crucial for a higher success avoiding parasite infection.