Phelipanche aegyptiaca management in tomato

Phelipanche and Orobanche species (broomrapes) are root holoparasitic plants that cause severe damage to economically important crops. Phelipanche and Orobanche spp. are widespread in Mediterranean areas, in Asia and in Southern and Eastern Europe, attacking dicotyledonous crops and depending entirely on their hosts for all nutritional requirements. Phelipanche aegyptiaca , Phelipanche ramosa and Orobanche cernua are extremely troublesome weeds on tomatoes. These noxious parasites exert their greatest damage prior to their shoot emergence and flowering; therefore, the majority of field loss may occur before diagnosis of infection. This review summarises the four main control measures for the weedy root parasites Phelipanche and Orobanche in tomato, namely chemical and biological control, resistant varieties and sanitation. Some of these methods are commercially widely used by farmers in Israel (chemical control), some are in the final stages of development towards commercialisation (resistant varieties and sanitation), and some still require further development and improvement before commercial implementation (biological control). The review presents an up-to-date summary of the available knowledge on their use for broomrape management in processing tomatoes.     

列当(Orobanche spp. and Phelipanche spp.)种子的采集与预处理方法

列当(Orobanche spp.and Phelipanche spp.)是一种在世界范围内危害严重的根寄生杂草,防除列当的研究也越来越多。正确地采集和预培养列当种子是获得正确防除列当研究结果的前提。列出了大田正确采集列当种子的方法,以及对采集到的种子进行过筛提纯、加洗洁精和吐温20清洗后,再表面消毒和预培养,并通过发芽试验检验此方法的可行性。结果表明,经过处理的向日葵列当、瓜列当种子的发芽率最高分别为54.4%、59.1%。此方法可以应用于列当的采集和预处理过程中,可为列当研究和防除提供参考。     

Evaluation of the pathogenicity of microorganisms isolated from Egyptian broomrape (Orobanche aegyptiaca) in Israel

Diseased Egyptian broomrape ( Orobanche aegyptiaca ) inflorescences were collected from a heavily broomrape-infested tomato ( Lycopersicon esculentum ) field in Israel. The microorganisms that were isolated from the diseased inflorescences were passed through Koch's postulates on Egyptian broomrape-parasitizing tomato roots in a polyethylene bag system and pots under greenhouse conditions. The fungi, Alternaria alternata , Macrophomina phaseolina , Rhizoctonia solani , and Fusarium solani , and the bacterium, Bacillus sp., were newly isolated from the diseased inflorescences of Egyptian broomrape and were found to be pathogenic to the parasite. Fusarium solani damaged all of the developmental stages of broomrape and prevented the damage that Egyptian broomrape causes to tomato plants. The level of pathogenicity and the damage of M. phaseolina , A. alternata , and Bacillus sp. to Egyptian broomrape in greenhouse experiments were relatively low. All the tested microorganisms are known as pathogens of tomato, yet none caused disease symptoms on the tomato plants grown in the inoculated polyethylene bags or in the pots. Fusarium solani demonstrated the highest potential for further development as a mycoherbicide for Egyptian broomrape control in tomato.     

烟草及向日葵上列当Orobanche cumana的发生及其生物防治

 近年,在河北省部分烟草和向日葵种植地发现大量寄生性杂草列当发生并造成严重为害。经鉴定为向日葵列当Orobanche cumana Wallr.。向日葵、烟草、番茄和黄瓜等作物的根分泌物能够刺激向日葵列当O. cumana种子萌发,而油葵根的分泌物则不能刺激向日葵列当种子萌发。利用列当致病镰刀菌L2菌株进行田间生物防治,防治效果达到92.4%。L2菌株对小麦、玉米、棉花、烟草和向日葵等作物生长无影响,L2菌株粗毒素能够抑制列当种子萌发,并造成列当种子萌芽管损伤。     

Influence of Root Exudates on Orobanche Seed Germination*

Broomrape (Orobanche ramosa L.) is a common root parasite of solanaceous, leguminous and other crops grown in the semi-arid regions of the world. The seeds germinate when root exudates from host plants are released in their immediate vicinity (Lindley, 1853; Koch, 1887; Chabrolin, 1934). Brown et al. (1951a) reported that non-host plants, such as flax (Linum usitatissimum L.) may stimulate Orobanche seed germination without being parasitized. The stimulating properties of flax exudate were studied by Brown et al. (1951b). They reported that the stimulant was unstable in alkaline solutions, but moderately stable in weakly acidic media, which may indicate the presence of an acidic (lactone) grouping. Nash & Wilhelm (1960) reported that gibberellic acid in agar media stimulated O. ramosa seed germination. Abu- Shakra, Miah & Saghir (1970) found that pre-treatment of 0. ramosa seeds with 100 ppm of gibberellic acid followed by incubation on a flax-root diffusate agar medium gave a high (81·7%) germination. The purpose of this study was to collect root exudates from three species of plants cultured under three experimental systems, namely (a) germ-free, (b) glasshouse (non-sterile), and (c) growth chamber (hydroponic, initially aseptic), and to evaluate their biological activity as germination stimulants for O. ramosa seeds. The plants used were tomato (Lycopersicon esculentum Mill.), sorghum (Sorghum vulgare Pers.) and flax. Exudate from marigold (Tagetes erecta L.) also was collected from germ-free culture.     

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.     

In vitro culture of Orobanche ramosa

Orobanche spp. (broomrapes) are holoparasites that subsist on the roots of many important crops and can considerably reduce yield. The control of Orobanche spp. includes physical, chemical and biological methods. Interactions between parasitic angiosperms and their hosts first occur at the level of parasite seed germination. The seeds of all Orobanchaceae germinate in soil under natural conditions only in response to specific chemical exudates from the host plant. This study describes the influence of different plant growth regulators and host plant root exudates on germination and development of calli from Orobanche seeds in vitro . The effect of indole-3-acetic acid, gibberellic acid and kinetin on the germination of Orobanche seeds varied with concentration. These plant growth regulators also affected the period of germination and the structure of calli and protrusions. An in vitro system for the collection of tobacco root exudates was established. Compounds released from the host roots of three different tobacco cultivars were found to provoke high levels of germination of the Orobanche seeds without any period of pre-conditioning. This study developed methods for the investigation of host–parasite interactions and the effect of germination stimulants in Orobanche spp.     

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.     

Investigation of wheat as a trap crop for control of Orobanche minor

Orobanche minor is a parasitic weed that attaches to the roots of red clover (Trifolium pratense) and a number of other broad‐leaved plant species in the Pacific Northwest USA. Orobanche minor seed must be stimulated by host plant exudates for germination and attachment to occur. However, plant species called false‐hosts can stimulate parasitic seed germination without attachment. These species could be utilized as trap crops to reduce the amount of parasitic seed in infested soil. Wheat (Triticum aestivum), was found to be a false‐host of O. minor; therefore, growth chamber, glasshouse and field soil experiments were conducted to evaluate the effect of six soft white winter wheats (T. aestivum), one durum wheat (Triticum turgidum), and one triticale (Triticale hexaploide) on O. minor germination. In growth chamber experiments, wheat and triticale induced 20–70% of O. minor seeds to germinate. In glasshouse studies, O. minor attachment was minimal on red clover plants grown in pots previously planted to wheat or triticale. In pots that did not receive a false‐host treatment, red clover plants averaged 4.2 O. minor attachments per plant. Red clover plants also had fewer O. minor attachments when grown in field soil taken from the plots where wheat or triticale were grown compared with plants grown in soil where no wheat or triticale were previously grown. Our results demonstrate that wheat may have the potential to be effectively integrated into an O. minor management system.     

Variation in resistance to Orobanche crenata in species of Cicer

Crenate broomrape (Orobanche crenata) is a major constraint for legume cultivation in Mediterranean agriculture. Field trials, pot and in vitro experiments demonstrated that resistance to O. crenata is present in chickpea and wild species of Cicer. The resistance is the result of the combination of several mechanisms, including low induction of parasite seed germination and in some accessions, either a darkening at the infection site on the host root that prevents establishment, or a reduced development of established parasite tubercles.     

A novel strain of Fusarium oxysporum from Germany and its potential for biocontrol of Orobanche ramosa

A newly isolated Fusarium oxysporum strain was investigated for its biocontrol potential against the root parasitic weed Orobanche ramosa (branched broomrape). The fungus was found to affect all developmental stages of the parasite. Orobanche seed germination was reduced by 40% in the presence of fungal conidia in vitro . The number of underground tubercles and shoots of the weed was reduced by the fungus by 55% compared with the control and 92% of tubercles were recorded as diseased in root chambers. In pot experiments, soil application of a granular formulation of the fungus resulted in a reduction of number and dry matter of Orobanche shoots by more than 90%. Spraying of a conidial suspension on aboveground Orobanche shoots caused the death of 75% of them within 2 weeks. Data from initial host-range experiments indicate that the isolate is very host-specific, not even attacking shoots of other Orobanche species. Because of these promising results, we conclude that the fungal isolate should be investigated under field conditions and be compared with other Fusarium isolates proposed for biocontrol of O. ramosa .     

Germination of Fusarium oxysporum in root exudates from tomato plants challenged with different Fusarium oxysporum strains

The response of microconidia from pathogenic and non-pathogenic Fusarium oxysporum to root exudates from tomato plants inoculated with different pathogenic and non-pathogenic F. oxysporum strains was studied. Root exudates from non-inoculated tomatoes highly stimulated the microconidial germination of the two tomato pathogens, F. oxysporum f.sp. lycopersici strain Fol 007 and F. oxysporum f.sp. radicis-lycopersici strain Forl 101587. In root exudates from tomato plants challenged with the pathogen Fol 007 the microconidial germination of Fol 007 was increased, whereas in root exudates from plants challenged with Forl 101587 the microconidial germination of Fol 007 was reduced. Root exudates of tomato plants challenged with the non-pathogenic unspecific F. oxysporum strain Fo 135 and the biocontrol strain Fo 47 clearly reduced microconidial germination of the pathogenic strain Forl 101587. Moreover, the microconidial germination rate of the biocontrol strain Fo 47 was increased in the presence of root exudates of tomato plants challenged with the tomato wilt pathogen Fol 007. These results indicate that pathogenic and non-pathogenic F. oxysporum strains alter the root exudation of tomato plants differently and consequently the fungal propagation of pathogenic and non-pathogenic F. oxysporum strains in the rhizosphere is affected differently.     

Effects of arbuscular mycorrhizal fungal symbiosis for control of Egyptian broomrape (Orobanche aegyptiaca Pers.) in tomato (Lycopersicon esculentum L.) cultivation

The present study assessed the effect of arbuscular mycorrhizal fungi (AMF) on broomrape (Orobanche aegyptiaca Pers.) control to determine its effects on tomato (Lycopersicon esculentum L.) growth indices. This experiment was carried out using a randomized complete block design with four replications. The treatments included three strains of AMF (Glomus mosseae, Glomus intraradices and Glomuse hoe ) at three levels (100, 200 and 300 kg ha^?1). The control treatments (without mycorrhiza) were with and without broomrape (weed infest and weed free). The results showed that the use of G. intraradices and G. mosseae decreased broomrape seed germination, the number of nodules and the dry weight of the broomrape and increased root area and dry weight of the tomato plant when compared to G. hoe and the control treatments without broomrape. The tomato yield in G. intraradices 300 kg ha^?1 treatment increased by 10 and 205% than weed free and weed infest treatments, respectively. Finally, G. intraradices and G. mosseae at, respectively, 200 and 300 kg ha^?1 are recommended for land under tomato cultivation that has been contaminated with broomrape.     

Seed viability determination in parasitic broomrapes (Orobanche and Phelipanche) using fluorescein diacetate staining

Fluorescein diacetate (FDA) was investigated as a potential viability stain for seeds of holoparasitic broomrapes Orobanche and Phelipanche (Orobanchaceae), using four weedy and two non-weedy taxa. FDA viabilities were compared with the currently used 2,3,5-triphenyl tetrazolium chloride (TTC) test, and in vitro germination induced by the strigolactone analogue GR-24. Viable FDA-stained seeds were characterised by bright yellow–green fluorescence under 450–490 nm blue light. These viable seeds could be discriminated from non-viable seeds, in which low levels of fluorescence, similar to non-stained control seeds were observed. FDA viabilities were not significantly different from those obtained using the TTC stain; however, viability values for both stains were consistently higher than levels of artificially induced germination. Positive TTC-staining showed continuous variation that was difficult to interpret. Nevertheless, the TTC test does not involve grinding seeds, and therefore probably remains a practical alternative to FDA for screening soil samples contaminated with Orobanche and Phelipanche seeds, which indicates different applications for the two viability tests. Interestingly, the strigolactone analogue GR-24 only induced germination in O. crenata , P. ramosa and O. minor , suggesting a high specificity towards this germination inducer among the species investigated. We suggest that FDA-staining provides a potential alternative to the currently used TTC test for seed viability assays for Orobanche and Phelipanche , and other parasitic weeds which are an obstacle to crop cultivation.     

Some compatible Rhizobium leguminosarum strains in peas decrease infections when parasitised by Orobanche crenata

Orobanche crenata (crenate broomrape) produces serious damage to many legume crops and particularly becomes a limiting factor for pea production in the Mediterranean basin. Nodulation effects on pea–broomrape relationships were studied using the commercial pea cultivar Douce de Provence and different Rhizobium strains using pot and Petri dish experiments. First, the benefit of bacterial inoculation on plant growth and efficiency of N incorporation were demonstrated for two isolates, P.SOM and P.1236. These isolates did not influence parasite germination induced by the artificial stimulant, GR24. In contrast, pea root inoculation with P.SOM and P.1236 isolates led to a reduced root infection by O. crenata , resulting from a lower Orobanche germination rate close to pea roots and a limited capacity of the parasitic seedlings to develop tubercles. Broomrape necrosis was observed both before and after parasite attachment to inoculated pea roots. Concomitantly, reduction in infection was accompanied by enhanced peroxidase activity and constantly high phenylalanine ammonia lyase activity in pea roots. These data suggest the involvement of these enzymes in pea resistance to crenate broomrape induced by the compatible rhizobia. Management of Orobanche via crop selection based on these enzyme systems is a viable option.     

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.     

Identification of sources of resistance to crenate broomrape (Orobanche crenata) in Spanish lentil (Lens culinaris) germplasm

The weedy root parasite Orobanche crenata constitutes a serious threat to lentils and other legumes grown in the Mediterranean and western Asia. Control strategies have centred on agronomic practices and the use of herbicides. Resistance breeding is hampered by scarcity of proper sources of resistance and of a reliable and practical screening procedure. A stepwise procedure was used here. A Spanish germplasm collection of 234 accessions of lentils (Lens culinaris) was screened for resistance to crenate broomrape under field conditions. A wide range of responses was observed, however, complete resistance was not detected. Some accessions exhibited a substantial reduction in broomrape infection. Thirty‐five lentil accessions with reduced broomrape infection were selected for further screening in the field and to determine the components of resistance in pot and in mini‐rhizotron experiments. Low infection seemed to be based on a combination of various escape and resistance mechanisms: (i) a lower root density, (ii) a lower induction of Orobanche seed germination, (iii) the reduced establishment of broomrape radicles and (iv) a limited development of established tubercles. In addition, necrosis of tubercles was observed in some accessions. This study shows how complementary in vitro screening methods may be used to rank and identify lentil accessions with potential broomrape resistance, based on different resistance mechanisms which may in the future be combined into a single cultivar by breeding.     

Intraspecific seasonal variation of dormancy and mortality of Phelipanche ramosa seeds

Phelipanche ramosa (Branched broomrape) is an obligate root parasitic plant that is a major pest of oilseed rape in France. Knowledge on seed viability and dormancy under field conditions is crucial to understand how to control P. ramosa, but is as yet unknown. Our study aimed to quantify these processes with a 2‐year seed burial experiment. Two genetically distinct populations of P. ramosa were studied, collected on winter oilseed rape (population O) and hemp (population H). Seed mortality was very low in both populations (4–7% per year). Although obligate parasitic seeds are assumed to germinate only after exposure to germination stimulants from host root exudates, a high proportion of population H seeds germinated spontaneously (up to 90%). Seeds of both populations displayed seasonal dormancy, with timing and magnitude depending on the population. Dormancy was low at the time each native host crop is usually sown. Populations differed in germination dynamics, with seeds of population H germinating faster. The difference in behaviour that we observed between populations is consistent with reported adaptations of pathovars to their preferred hosts. The results indicate that the parasitic plant management requires targeting at the populations concerned. For example, delayed sowing is more promising against population O than against population H.     

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.     

Increased rhizosphere populations of Trichoderma asperellum strain T34 caused by secretion pattern of root exudates in tomato plants inoculated with Botrytis cinerea

Root exudates secreted from plants can modify rhizosphere microbiota by enhancing or inhibiting the growth of biological control agents (BCAs) and/or pathogens. Similarly, microorganisms can modify the secretion of plant root exudates. The aim of this study was to analyse the effect of a Botrytis cinerea leaf infection on the secretion of tomato root exudates and on the populations of the BCA Trichoderma asperellum strain T34 (T34). This study found that the secretion pattern of root exudates in tomato plants was influenced by B. cinerea infection in plant leaves. An increase in the levels of gluconic acid was observed, while levels of sucrose and inositol decreased. A decrease in the severity of B. cinerea by the induction of systemic resistance triggered by T34 was also observed. Tomato plants infected with B. cinerea maintained the populations of T34 in the roots, while populations of T34 decreased in plants not inoculated with the pathogen. Samples exposed to media containing gluconic acid (as the only carbon source or at the same concentration found in roots exudates) saw an increase in the in vitro growth of T34 compared to media without gluconic acid. In conclusion, a change in the secretion pattern of root exudates caused by B. cinerea, together with the enhanced growth of T34 in the presence of gluconic acid, indicates the existence of leaf to root communication. The result of this is enhanced populations of T34, and in turn induced disease resistance and a consequential reduction in disease severity.