Fenugreek root exudates show species-specific stimulation of Orobanche seed germination

Various Orobanche species are weedy and cause severe reduction in the yields of many important crops. The seeds of these parasitic weeds may remain dormant in the soil for many years until germination is stimulated by the release of a chemical signal from a host plant. In order to determine the effects of fenugreek root exudate on the induction of Orobanche crenata, Orobanche ramosa and Orobanche foetida seed germination, root exudate was collected from hydroponically grown fenugreek seedlings. Fractionation patterns obtained from column and thin layer chromatography of the fenugreek root exudate showed a set of metabolites differing in their polarity with stimulatory activity on Orobanche seed germination. The crude root exudate stimulated both O. ramosa and O. crenata seed germination to the same level caused by the synthetic germination stimulant GR24 at 10 mg L^?1. It also stimulated O. foetida seed germination which did not respond to GR24. Active fractions of root exudate stimulated the germination of Orobanche species differentially.     

Biology and management of the fast‐emerging threat of broomrape in rapeseed–mustard

Broomrapes (Phelipanche and Orobanche spp.) are obligate root parasites of the family Orobanchaceae. The natural variation in Orobanchaceae exists in plants involving Triphysaria versicolor (facultative hemiparasite), Striga hermonthica (obligate hemiparasite) and Phelipanche aegyptiaca, formerly Orobanche aegyptiaca (holoparasite). The family Orobanchaceae has 90 genera. The four major parasitic species of broomrape are Phelipanche ramosa in the Brassicas, especially in rapeseed, Phelipanche cernua in tobacco and sunflower, P. aegyptiaca in solanacious crops and in Brassica juncea and Phelipanche crenata in leguminous crops. They are prevalent, infesting nearly 1.2% of the world's arable land. In India, P. ramosa and P. aegyptiaca cause severe infestations of Brassica and have threatened its cultivation in major growing areas. In addition, a single broomrape plant can release >100 000 seeds that remain viable for decades in the soil. This provides the parasite with a great genetic adaptability to environmental changes, including host resistance, agronomical practices and herbicide treatments. Different control measures, such as manual weeding, herbicide applications, solarization, crop rotation and integrated broomrape management practices, have been proposed in order to reduce the broomrape menace and improve yields in farmers' fields. Breeding for broomrape resistance also could be an economic, feasible and environmental friendly control method. The present article reviews the current status of research on broomrape in India and abroad, as well as suggests strategies for its effective management.     

Characterization of the Orobanche–Medicago truncatula association for studying early stages of the parasite–host interaction

There is an increasing interest in the legume species Medicago truncatula as a model plant for structural and functional genomic studies that can be used to identify agronomically important genes in legumes. Field screening has shown very high levels of resistance to Orobanche crenata in M. truncatula. However, in vitro studies with O. crenata, Orobanche foetida, Orobanche ramosa and Orobanche minor showed useful variation among accessions at early stages of the parasite–host interaction. Significant differences were observed in the levels of germination of O. crenata and O. foetida seeds induced by different accessions of M. truncatula. Only limited germination was observed on accession SA‐4327. All accessions induced little O. ramosa and O. minor germination. Accessions also varied in the number of O. crenata and O. foetida attachments supported, with few developing on accession SA‐27774. The variation observed for induction of germination and of subsequent attachment will be useful to isolating and characterizing genes involved in the early stages of Orobanche–host plant interaction and for the study of the biosynthetic pathways of production for germination stimulants.     

Use of ITS nuclear sequences from Phelipanche aegyptiaca as a direct tool to detect single seeds of broomrape species in the soil

Broomrape (Phelipanche and Orobanche spp.) are obligate holoparasites that attack roots of almost all economically-important crops in semiarid regions of the world. Broomrape seeds are extremely small (dust-like seeds), averaging 200 to 300?μm in size and because of the miniscule seed size it is difficult to detect and confirm via conventional methods. In this study our aim was to develop a PCR-based assay specific for broomrape soil-borne seeds and sensitive enough to detect a single or few broomrape seeds in a soil sample. For this purpose, we used complementary polymerase chain reaction (PCR) primers based upon unique sequences in the internal transcribed spacer (ITS) regions of the nuclear ribosomal DNA of Phelipanche aegyptiaca. Genomic DNA was extracted from soil samples artificially infested with broomrape seeds or tissue of Phelipanche aegyptiaca Pers., Orobanche cumana Wallr. and Phelipanche crenata Forsk. and subjected to PCR analysis. Using ITS-350 primers, a specific PCR product (350?bp) was amplified and detected in all samples containing broomrape species, but was not detected in soil sample free of broomrape seeds or tissues. Additionally, the PCR-based assay was sensitive enough to detect even a single broomrape seed in the soil. As expected the universal internal control primers amplified a PCR product (555?bp) of genomic DNA extracted from soil samples with or without broomrape tissues or seeds. This diagnostic method is simple, reliable and rapid and could help for assessment of broomrape seed contamination in a crop field.     

Biochemical factors involved in vetch resistance toOrobanche aegyptiaca

Experiments were performed to elucidate the biochemical nature of the differential response of resistantVicia atropurpureacv. Popany and susceptibleVicia sativacv. Yovel to the parasitic weedOrobanche aegyptiaca. Plant root material was obtained by growing vetch plants in association withOrobancheseeds on glass microfibre filter sheets inserted in polyethylene bags replenished by nutrient solution. Higher concentrations of bound phenolics, free phenolics and lignin, and higher activity of peroxidase were observed in cv. Popany roots as a result ofO. aegyptiacainfection than were found in cv. Yovel vetch. No differences in hydroxyproline-rich glycoproteins levels were detected between infected and non-infected roots of either vetch cultivar. These results suggest that the resistant vetch defense mechanism involves elevated induction of the phenylproponoid pathway uponOrobancheinfection, conferring mechanical and chemical barriers confronting the invading parasite.     

Broomrape (Orobanche spp.) control with ethylene dibromide and chloropicrin

Ethylene dibromide effectively controlled Orobanche crenata Forsk. and O. cernua Loefl. but was not effective against O. aegyptiaca Pers. and O. mutelii F. Schultz. Chloropicrin was considerably less effective than ethylene dibromide. Mixtures of both compounds did not result in improved broomrape control but nonetheless had a positive effect on crop yield. Destruction des Orobanches avec du dibromure d'ethylène et de la chloropicrine Le dibromure d'éthylène a été efficace contre Orobanche crenata Forsk et O. Cernua Loefl, mais non efficace contre O. aegyptiaca Pers, et O. mutelii F. Schultz. La chloropicrine est beaucoup moins efficace que le dibromure d'ethylène. Les mélanges des deux matières actives n'ont pas amélioré l'efficacité sur orobanches mais néanmoins ont eu un effet positif sur le rendement de la culture. Bekämpfung von Orobanche-Arten mit Ethylendi-bromid und Chlorpikrin Mit Ethylendibromid konnten Orobanche crenata Forsk. und O. cernua Loefl., aber nicht O. aegyptica Pers. und O. mutelii F. Schultz bekämpft werden. Chlorpikrin war beträchtlich weniger wirksam. Mischungen der beiden Wirkstoffe ergaben keine bessere Wirkung, beeinflussten aber den Ertrag der Kulturpflanzen positiv.     

Germination response of Orobanche seeds subjected to conditioning temperature, water potential and growth regulator treatments

Broomrapes (Orobanche spp.) are parasitic weeds that cause significant losses of crop yield. Experiments were conducted to investigate the seed response to the artificial germination stimulant GR₂₄ in three species of Orobanche subjected to preconditioning under various temperatures, water potentials and with plant growth regulators. The highest germination percentages were observed in Orobanche ramosa, Orobanche aegyptiaca and Orobanche minor seeds conditioned at 18°C for 7 days followed by germination stimulation at 18°C. With the increase of the conditioning period (7, 14, 21 and 28 days), the germination percentage of O. ramosa and O. aegyptiaca progressively decreased. When conditioned at −2 MPa, the germination percentage was lower than at 0 and −1 MPa, especially at 13 and 28°C. Orobanche minor seeds could retain relatively high germination if conditioned at 18, 23 or 28°C, even after significantly extended conditioning periods (up to 84 days). GA₃ (30–100 mg L⁻¹), norflurazon and fluridone (10–100 mg L⁻¹), and brassinolide (0.5–1.0 mg L⁻¹) increased seed germination, while 0.01 mg L⁻¹ uniconazole significantly reduced germination rates of all three Orobanche spp. The promotional effects of GA₃ and norflurazon and the inhibitory effect of uniconazole were evident, even when they were treated for 3 days. Germination of Orobanche seeds was much lower when the unconditioned seeds were directly exposed to GR₂₄ at 10⁻⁶ m . This early GR₂₄-induced inhibition was however alleviated or even eliminated by the inclusion of GA₃ or norflurazon (10–50 mg L⁻¹) in the conditioning medium. On the contrary, the inclusion of uniconazole increased the inhibitory effect of GR₂₄, particularly in the case of O. ramosa.     

Ausbreitung des parasitischen Unkrauts Phelipanche ramosa in der deutschen Landwirtschaft

The root parasitic weeds Phelipanche ramosa (branched broomrape) and P. aegyptiaca have the widest host range among Orobanche and Phelipanche species. In Western Europe, P. ramosa attacks, with increasing aggressiveness, crops such as oilseed rape, tobacco, hemp, and tomato. The unique biology of root parasites, establishes a closed link with their host plant, thus reducing the possibility to successfully control them. Control measures include (a) physical processes (such as weeding, solarization, deep ploughing, burning off, flooding), (b) chemical (like soil fumigation, use of herbicides, germination stimulants) and (c) biological methods (e.g. resistant varieties, use of fungi and insects as antagonists, trap and catch crops). German tobacco growers rely mostly on the herbicide method. They apply glyphosate in very low concentrations, when the first tubercles are formed. Also a fungal antagonist against the parasitic weed on tobacco was found in Germany, but until now has not been developed into a commercial mycoherbicide. After hemp production lost its significance as a crop in Germany, tobacco remained as the main host for P. ramosa. In the past 10 years, branched broomrape has spread out and currently it can be found in areas where previously were free of it. Since the elimination of EU subsidies, some tobacco growers began to cultivate on their land parsley instead of tobacco. As a result, parsley has now been infected with P. ramosa. When used 10 years ago as catch crop, parsley had a rather small effect on branched broomrape. This potential danger, especially by other potential host plants, such as oilseed rape, tomato and potato or even weeds should not be underestimated. Spread and expansion of the host plant spectrum of branched broomrape in Germany might be reduced by the introduction of appropriate phytosanitary measures and improved information policies.     

Stimulating<Emphasis Type="Italic">Orobanche ramosa</Emphasis> seed germination with an<Emphasis Type="Italic">Ascophyllum nodosum</Emphasis> extract

Algit Super®, an extract ofAscophyllum nodosum, was found to be an effective stimulant for the germination ofOrobanche ramosa seeds. The biological activity of Algit Super® was tested at a wide range of concentrations from 2.5 v/v to 1.2 × 10^?3 v/v. The seed germination in all studiedO. ramosa populations showed a concentration dependence, with an inhibitory effect at higher concentrations. The response ofO. ramosa to Algit Super® resembled its response to the reference stimulant GR24. VariousO. ramosa populations demonstrated differential response to the stimulant.     

Identification,characterisation and discriminatory power of microsatellite markers in the parasitic weed Orobanche cumana

Orobanche cumana is an obligate root parasite of sunflower. It represents a major agricultural problem in many countries of southern and eastern Europe. Information on O. cumana population genetics, structure and dynamics is scarce, particularly due to the lack of suitable molecular markers for such studies. The objective of this study was to identify and characterise simple sequence repeat (SSR) markers for O. cumana. Four thousand two hundred SSR‐containing candidate sequences were obtained from O. cumana using next‐generation sequencing, from which 298 SSR primer pairs were designed and 217 of them used for validation. Seventy nine SSR primers produced reproducible, high quality amplicons of the expected size that were polymorphic among 18 O. cumana populations from different geographical locations and hosts (sunflower, wild hosts from the Compositae family). The number of alleles per locus ranged from 2 to 10, with an average polymorphism information content value of 0.37. The O. cumana SSR markers were highly transferable to the closely related species Orobanche cernua. SSR markers showed high resolving power; UPGMA cluster analysis allowed proper classification of Orobanche spp. samples into species (O. cumana and O. cernua), geographical origin and host. The functional SSR markers reported in this study constitute a valuable tool for genetic analyses in O. cumana and related species and will contribute insights into the biology and genetics of this parasitic weed.     

Prediction of yield loss caused by Orobanche spp. in carrot and pea crops based on the soil seedbank

Root parasites of the genus Orobanche cause serious losses in many subtropical crops. Direct control options are very limited and crop yield loss can reach 100%. Prediction of potential damage in a crop before sowing or planting would support farmers in their choice of crop. This paper discusses the relationship between the number of Orobanche spp. seeds in the field and yield loss in peas (Pisum sativum L.) and carrots (Daucus carota L.) in Israel. Yield loss due to Orobanche crenata Forsk. in peas was 100% at high infestations, whereas in carrots when O. crenata and O. aegyptiaca were present it stabilized at about 50% for moderate infestations of 200 seeds per kg of soil. Statistical analyses related the yield loss from parasitism in peas and carrots to the numbers of Orobanche seeds remaining in the soil. A rectangular hyperbolic model, previously applied to competition data, fitted the data well. Confidence intervals for per cent yield loss were calculated using the bootstrap method. The practical applications of these models in predicting yield loss are discussed.     

Nonspecific toxins as components of a host‐specific culture filtrate from Fusarium oxysporum f. sp. cubense race 1

Bananas and plantains (Musa spp.) are among the most important crops in the world providing staple food for hundreds of millions of people. However, banana production has been devastated by fungal infestations caused by Fusarium oxysporum f. sp. cubense (Foc). Despite the fact that there is very little known on the role of microbial metabolites in the molecular mechanism of Foc infections, it has been proposed that the toxins fusaric acid and beauvericin produced by Foc play an important role during pathogenesis. The aim of this contribution was to study the toxic components of culture filtrates (CF) of Foc and to isolate the extracellular microbial metabolites involved in the plant response. An in vitro bioassay was used to evaluate the production of phytotoxic metabolites as well as the specificity of culture from a strain of Foc belonging to VCG 01210 (race 1). A host‐specific CF was obtained and the phytotoxic compounds characterized as fusaric acid, beauvericin and fumonisin B1. Despite the presence of these nonspecific toxins, a water‐soluble extract from the CF induced protection to the main phytotoxic fraction, measured by lesion area. This hydrophilic fraction induced a fast and strong response of just jasmonic acid (JA)‐dependent defence genes rather than salicylic acid (SA)‐ and ethylene (ET)‐response genes in resistant cultivars. Extracellular proteins isolated from CF of Foc provide an important source for further investigations on the molecular basis of the interaction between Foc and banana.     

Control of morningglory species using Fusarium solani and its extracts

Abstract

Natural products of an isolate of Fusarium solani App. & Wr. NRRL 18883 grown on rice medium were discovered to be phytotoxic to morningglory. This isolate was evaluated for biocontrol potential on morningglory species including ivyleaf [Ipomoea hederacea (L.) Jacq.], multicoloured (I. tricolor Cav.), paimleaf (I. wrightii Gray), pitted (I. lacunosa L.), purple moonflower (I. turbinata Lag.), red (I. coccinea L.), sharppod (I. cordatotriloba Dennstedt), smallflower (Jacquemontia tamnifolia (L.) Griseb], and tall [I. purpurea (L.) Roth] morningglory. When sprayed at a concentration of 10 g fungus‐infested rice per 50 ml of water, this isolate caused phytotoxic damage including necrosis, chlorosis, growth inhibition, and mortality. Deleterious effects were recorded in all species, ranging from necrotic spots to death, depending on the species. Mortality ranged from 0% at 3 weeks for purple moonflower to 89% for smallflower morningglory. Soil‐drench application (10–20 ml fungal material per 150 g soil) caused wilting and death. Root lengths of 7‐ and 10‐day‐old plants were reduced between 19 and 89% by soil‐drench application and 17–84% by spray application. Whole plant length reduction of 7‐ and 10‐day‐old plants ranged from 0 to 96%, and 39 to 96% for soil‐drench and spray applications, respectively. Testing of F. solani on weed and crop species showed that most broadleaved species were sensitive but monocotyledons were immune. Tests of the phytotoxic extract for known phytotoxins such as common trichothecene (deoxynivalenol) and non‐trichothecene compounds (fusaric acid, moniliformin, fumonisins) were negative. This is the first report of the use of F. solani against morningglory.     

The influence of growth conditions on biomass, toxins and pathogenicity of Fusarium oxysporum f. sp. orthoceras, a potential agent for broomrape biocontrol

The influence of different combinations of illumination and shaking on the growth dynamics, pathogenicity and toxin production of the fungus Fusarium oxysporum f. sp. orthoceras, a biocontrol agent of Orobanche cumana, was studied. The fastest biomass accumulation was obtained under shaking, with or without illumination, with the highest biomass obtained after 3–4 weeks of growth. The biological activity of chloroform extracts of the culture filtrate was characterised: it contained at least two main toxic metabolites that caused necrosis and wilting of various plants and led to mortality of germinating seeds of O. cernua, O. aegyptiaca, O. ramosa and O. cumana. The highest toxic activity of the chloroform extract was obtained under illumination without shaking after 3–4 weeks of growth. The two toxic metabolites were purified and identified as fusaric acid (FA) and 9,10‐dehydrofusaric acid (DFA). Both FA and DFA production began in the first week of growth, increasing gradually to their maxima after 4 weeks. The highest level of pathogenic activity of the fungus was obtained after three or more weeks of fungal growth. It can be concluded that in order to produce high levels of toxin and pathogenic activity, the fungus should be grown under illumination without shaking for 4 weeks.     

The role of peroxidase in the resistance of sunflower against Orobanche cumana in Russia

We present the results of a histochemical study comparing seedlings of races C and D of Orobanche cumana Wallr. (syn. O. cernua Loefl.) attacking sunflower (Helianthus annuus L.) in southern Russia. Three groups of O. cumana seedlings were distinguished according to the peroxidase content of the cells in the radicles: (1) those with neither extracellular nor intracellubr peroxidase and whose radicles have a smooth apex (these were classified as non-infective): (2) those with a high peroxidase content of the nuclei and the cytoplasm layer adjacent to the cell wall, as well as excretion of peroxidase from the apex of the radicles: (3) those with a similarly high peroxidase activity in the parasite cells, but without extracellular excretion. The apices of the radicles of the last two groups are swollen. It is suggested that these belong to O. cumana races C and D respectively. The extracellular peroxidase in O. cumana race C reacts with phenolic compounds, which are lignin precursors of the host, resulting in host resistance due to the formation of lignin layers in sunflowers possessing the Or3 gene for resistance. The absence of extracellular peroxidase in O. cumana race D prevents lignin formation and enables the parasite to attach to the host vascular system. Comparison of these data with the information on the earlier O. cumana races A and B, and older sunflower cultivars, points to a crucial role of peroxidase in the process of breeding new sunflower cultivars and the evolution of new O. cumana races.     

A dominant avirulence gene in Orobanche cumana triggers Or5 resistance in sunflower

Orobanche cumana is a weed that grows as a root parasite on sunflower. In general, the O. cumana–sunflower parasitic system is regarded to follow the gene‐for‐gene model, although this has never been demonstrated at the genetic level in O. cumana. The Or5 dominant gene in sunflower confers resistance to O. cumana race E, but not to race F. The objective of this research was to study the inheritance of avirulence/virulence in crosses between plants of O. cumana lines classified as races E and F. Four race E and three race F lines were developed, from which four race E × race F cross‐combinations were made, in three cases including reciprocals. In all cases, F₁ seeds did not have the ability to parasitise sunflower line P‐1380 carrying the Or5 gene, indicating dominance of race E avirulence allele(s). Five F₂ populations comprising a total of 387 F_2:3 families were evaluated on sunflower line P‐1380. In all cases, one‐fourth of the F_2:3 families did not possess the ability to parasitise P‐1380 plants, suggesting that race E avirulence and race F virulence on P‐1380 are allelic and controlled by a single locus. This study demonstrated the gene‐for‐gene interaction in the O. cumana–sunflower parasite system and provided useful information to identify genes involved in O. cumana virulence. The approach followed in this research can contribute to define precisely races of the parasite on the basis of the presence of avirulence genes.     

Orobanche crenata resistance and avoidance in pea (Pisum spp.) operate at different developmental stages of the parasite

Orobanche crenata (broomrape) is an important constraint to pea (Pisum sativum) cultivation in the Mediterranean area, because little resistance is available in commercial crop varieties. Field experiments have demonstrated that some resistance is present in a number of P. sativum and P. fulvum accessions. The goal of this work was to characterize such resistance. The Pisum–O. crenata interaction and the resistance symptoms were studied under controlled conditions by using Petri dish and polyethylene bag assays. The content of phenolics and peroxidase activity in host tissue from infected and non-infected plants were also measured. Resistance and avoidance mechanisms, acting at different developmental stages of the parasite, have been identified, including low stimulation of O. crenata seed germination, unsuccessful penetration of host roots, delay in post-attachment tubercle development and necrosis of the attached tubercles. Infection caused an increase in the content of total soluble phenolics in some Pisum genotypes. Peroxidase activity was higher in resistant than in susceptible accessions. Results obtained with different Pisum genotypes showed that resistance is the result of several mechanisms acting at different stages of the infection process. Resistance is also related to increased levels of peroxidase activity in host roots.     

A competition index for Orobanche crenata Forsk effects on broad bean (Vicia faba L.)

The influence of broomrape (Orobanche crenata Forsk) infection levels on commercial broad bean (Vicia faba L.) crops was investigated and an equation relating yield losses to O. crenata density was derived. O. crenata growth was weakly and negatively related to the final height or shoot number of the broad bean plant. O. crenata decreased seed yield mainly by reducing the number of pods. The number of seeds per pod was also decreased by O. crenata infection when competition occurred at late-crop growth stages when most of the pods had already developed. Bean unit seed weight was only weakly affected by infection. Correlation coefficients between O. crenata number and broad-bean seed yield per plant were significant at P=0·05 and greater than 0·70 in most locations. On average, about four O. crenata per broad-bean plant decreased seed yield by half. An estimation of the percentage of losses in any field can be calculated from the equation: %loss = 100 × 0·124 × OcN. where OcN is the average final number of emerged O. crenata per plant.     

Broomrape control in faba bean (Vicia faba L.) with glyphosate and imazaquin

Field studies were carried out to evaluate the efficacy of imazaquin and glyphosate in controlling broomrape (Orobanche crenata Forsk. and O. aegyptiaca Pers.) in faba bean (Vicia faba L.) in Northern Syria. Several rates and times of application of the herbicides were tested. Nearly complete control of broomrape was achieved with both the herbicides when they were sprayed twice (when the broomrape attachments to the faba bean roots were at tubercle stage of develoment and 15 days later) at a rate of application of 80 g a.i. ha^?1 for glyphosate and 10 g a.i. ha^?1 for imazaquin. However, the yield of faba bean did not increase with these treatments, suggesting that the herbicides at these rates were still causing some physiological disorder in the plant that counteracted the beneficial effect of broomrape control on the performance of faba bean plants.     

Granular formulation of Fusarium oxysporum for biological control of faba bean and tomato Orobanche

BACKGROUND: Orobanche spp. represent a serious threat to a wide range of crops. They are difficult targets for herbicides, and biological control could provide a possible solution. This work therefore aimed to formulate mycoherbicides of Fusarium with adequate shelf life and virulence against Orobanche but safe to faba bean and tomato. RESULTS: Only two isolates of Fusarium oxysporum Schlecht. (Foxy I and Foxy II) obtained from diseased Orobanche shoots were found to be pathogenic to Orobanche crenata Forsk. and Orobanche ramosa L. Conidial suspension of both isolates significantly decreased germination, attachments and tubercles of Orobanche. Microconidia and chlamydospores of both isolates were formulated as mycoherbicides encapsulated in a wheat flour–kaolin matrix (four different formulations). All formulations greatly diminished Orobanche emerged shoots, total shoot number, shoot height, attachment of emerged shoots, the germinated seeds that succeeded in emerging above the soil surface and dry weight. Meanwhile, disease incidence and disease severity of emerged shoots were enhanced. The shelf life was adequate, particularly for coarse, freshly prepared, low‐temperature‐stored, microconidia‐rich formulations. The induced growth reduction of Orobanche‐infected host plants seemed to be nullified by formulations, particularly at the highest dose. CONCLUSION: These formulations seemed to destroy Orobanche but appeared harmless to host plants. Hence, they could be efficiently used as mycoherbicides for biological control of Orobanche in faba bean and tomato. Copyright © 2008 Society of Chemical Industry