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.     

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.     

Phenology and growth of Orobanche crenata Forsk (broomrape) in four legume crops

Studies were conducted in the field in 2 years comparing the phenology and growth of Orobanche crenata (Forsk) (crenate broomrape) in lentils (Lens esculenta L.) cv. Castellana, peas (Pisum sativum L.) cv. Orix, vetch (Vicia sativa L.) cv. comun and broadbean (Vicia faba L. cv. Alameda. First attachment of O. crenata to these crops took place 9–14 weeks after mid-November planting and earlier after later plantings, Differences in the first O. crenata attachment dates and in the duration of the underground growth period of O. crenata were much greater between years for any given crop than between crops in a given year: both attributes were apparently affected more by seasonal climatic conditions than by crop species, Furthermore, there was no consistent relationship found between crop growth stages and time after first attachment of the parante. This occurred in lentils and vetch while they were vegetative, in peas at late vegetative-early flowering stages, and at widely varying growth stages in broadbean, depending on planting dates and years. The maximum number of O. crenata plants successfully attached to each individual crop plant decreased in the order: peas > broadbean > lentil > vetch, with 21, 14, 10 and 8 per plant, respectively, averaged over the two seasons. Similarly, plant parasites: host dry weight ratio were 1.0, 0.7, 0.3 and 0.2 For each of these crops, respectively.     

A Greenhouse Test for Screening Sugar Beet (Beta Vulgaris) for Resistance to Rhizoctonia Solani

Rhizoctonia solani Kühn is a serious plant pathogenic fungus, causing various types of damage to sugar beet (Beta vulgaris L.). In Europe, the disease is spreading and becoming a threat for the growing of this crop. Plant resistance seems to be the most practical and economical way to control the disease. Experiments were carried out to optimise a greenhouse procedure to screen plants of sugar beet for resistance to R. solani. In the first experiment, two susceptible accessions were evaluated for root and leaf symptoms, after being grown in seven different soil mixtures and inoculated with R. solani. The fungus infected all plants. It was concluded that leaf symptoms were not reliable for the rating of disease severity. Statistically significant differences between the soil mixtures were observed, and there were no significant differences between the two accessions. The two soil mixtures, showing the most severe disease symptoms, were selected for a second experiment, including both resistant and susceptible accessions. As in the first experiment, root symptoms were recorded using a 1–7 scale, and a significant expression of resistance was observed. The average severity of the disease in the greenhouse experiment generally was comparable with the infection in field experiments, and the ranking of the accessions was the same in the two types of experiments. It was concluded that evaluation procedures in the greenhouse could be used as a rapid assay to screen sugar beet plants for resistance to R. solani.     

Host status and reaction of <Emphasis Type="Italic">Medicago truncatula</Emphasis> accessions to infection by three major pathogens of pea (<Emphasis Type="Italic">Pisum sativum</Emphasis>) and alfalfa (<Emphasis Type="Italic">Medicago sativa</Emphasis>)

Ditylenchus dipsaci, the stem nematode of alfalfa (Medicago sativa), Mycosphaerella pinodes, cause of Ascochyta blight in pea (Pisum sativum) and Aphanomyces euteiches, cause of pea root rot, result in major yield losses in French alfalfa and pea crops. These diseases are difficult to control and the partial resistances currently available are not effective enough. Medicago truncatula, the barrel medic, is the legume model for genetic studies, which should lead to the identification and characterization of new resistance genes for pathogens. We evaluated a collection of 34 accessions of M. truncatula and nine accessions from three other species (two from M. italica, six from M. littoralis and one from M. polymorpha) for resistance to these three major diseases. We developed screening tests, including standard host references, for each pathogen. Most of the accessions tested were resistant to D. dipsaci, with only three accessions classified as susceptible. A very high level of resistance to M. pinodes was observed among the accessions, none of which was susceptible to this pathogen. Conversely, a high level of variation, from resistant to susceptible accessions, was identified in response to infection by A. euteiches.     

Vavilov wheat accessions provide useful sources of resistance to tan spot (syn. yellow spot) of wheat

Host genetic resistance is the most effective and sustainable means of managing tan spot or yellow spot of wheat. The disease is becoming increasingly problematic due to the adoption of minimum tillage practices, evolution of effector‐mediated pathogenicity, and widespread cultivation of susceptible cultivars from a narrow genetic base. This highlights the importance of broadening the diversity of resistance factors in modern breeding germplasm. This study explored 300 genetically diverse wheat accessions, originally sourced from the N. I. Vavilov Institute of Plant Genetic Resources (VIR), St Petersburg, Russia. The collection was screened for resistance to tan spot at seedling and adult stage under controlled conditions, and in the field across 2 years. The phenotypic datasets, coupled with ToxA bioassay screening, identified a number of accessions with useful sources of resistance. Seedling disease response corresponded well with ToxA sensitivity (r = 0.49, P < 0.000), but not adult responses (r = ?0.02 to ?0.19, P < 0.002), and overall reactions to ToxA appeared to show poor correspondence with disease response at the adult stage. ToxA‐insensitive accessions were generally found resistant across different growth stages (all‐stage resistance, ASR) in all experiments (seedling and adult stage under controlled conditions and field). ToxA‐sensitive accessions that were susceptible at seedling stage, but resistant at both adult‐plant stages, were deemed to carry adult‐plant resistance (APR). This study provides detailed information on the degree of tan spot resistance in the Vavilov wheat collection and discusses strategies to harness these sources to boost the diversity of resistance factors in modern wheat breeding germplasm.     

Evaluation of cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis>) cultivars grown in Eastern Europe and progress in breeding for resistance to angular leaf spot (<Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv. <Emphasis Type="Italic">lachrymans</Emphasis>)

Increased occurrence of cucumber angular leaf spot, Pseudomonas syringae pv. lachrymans, has caused significant losses in cucumber, Cucumis sativus, yield in Poland in recent years. These losses necessitated evaluation of the level of resistance in cucumber cultivars of mainly Polish breeding, cultivated in Eastern Europe, and initiation of a breeding programme for resistance to this disease. Screening for resistance was performed on 84 cucumber accessions under growth chamber conditions using a highly aggressive strain of P. syringae pv. lachrymans. Most of the screened accessions were either susceptible or displayed intermediate resistance. The screening resulted in the identification of five F₁ hybrid cultivars moderately resistant to angular leaf spot. The identified F₁ hybrids were self-pollinated up to the F₄ generation. Individuals resistant to angular leaf spot were identified. These individuals can be used as a source of resistance to angular leaf spot in future breeding efforts.     

Resistance to blast (Magnaporthe grisea) in a mini-core collection of finger millet germplasm

Blast caused by Pyricularia grisea [teleomorph: Magnaporthe grisea] is an economically important and widespread disease of finger millet in the world. Host resistance is the most economical and effective means of combating this disease as finger millet is predominantly grown by resource-poor and marginal farmers. At the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), we evaluated a finger millet mini-core collection of 80 germplasm accessions (about 1 % of the total germplasm collection representing major trait variability) for blast resistance both in the field and greenhouse. Field evaluation was done using a refined screening technique that included new improved rating scales for leaf, neck and finger infection. Sixty six of the 80 accessions showed combined resistance to leaf, neck and finger blast in two seasons (2009 and 2010) of field screening. A highly significant and positive correlation was found between neck and finger blast ratings (r?=?0.92), whereas small but significant correlations were found between leaf blast and neck blast (r?=?0.25) and between leaf blast and finger blast (r?=?0.30). These accessions were also screened for leaf blast resistance in the greenhouse by artificial inoculation of seedlings to confirm field observations. Fifty-eight of the 80 accessions were resistant to leaf blast in the greenhouse screen as well. These resistant accessions represented one wild (africana) and four cultivated races (vulgaris, plana, elongate and compacta) of finger millet that originated from 13 countries in Asia and Africa and exhibited considerable diversity for agronomic traits, such as maturity period, plant height and panicle type. These blast resistant accessions from the mini-core collection would be useful in finger millet disease resistance breeding programs.     

An in-vitro growth system for studying the parasitism of cowpea (Vigna unguiculata) by Striga gesnerioides

An in-vitro growth system for investigating the behaviour of S. gesnerioides on cowpea has been developed. The roots of young cowpea plants were spread over glass fibre filter paper in a shallow plastic tray, and a known number of germinated Striga seeds were placed on the cowpea root surface. This allowed the infection process of the parasite to be quantified. Good access to the host-parasite association enabled the entire infection process to be easily monitored and tissue removed for cytological studies. Observations revealed for the first time that contact of S. gesnerioides radicles with host roots stimulated the development of radicular hairs and swelling of the tip of these radicles. Furthermore, penetration of cowpea roots by Striga was shown to be localized and to cause very little damage to the host roots. Swollen parasite tubercles developed on the surface of the roots only after the host stele had been penetrated by invading Striga cells. This growth system is suitable for in-vestigating the mechanisms of host resistance. and also for rapidly screening cowpea germplasm for new sources of resistance to the Parasite. Resistant cowpea cultivars offer the best potential solution for control of this Parasite.     

Root hair infection by Plasmodiophora brassicae in clubrootresistant and susceptible genotypes of Brassica oleracea,B. rapa and B. napus

The pathogenesis of clubroot, a disease of cruciferous crops caused by the fungusPlasmodiophora brassicae, starts with infection of the root hairs. This process was studied in 13 accessions ofBrassica oleracea, B. napus and B. rapa with varying levels of plant resitance toP. brassicae. Seedlings were grown in a mineral solution, inoculated with resting spores ofP. brassicae, and the number of plasmodia developing in root hairs was recorded. When compared with the standard susceptible cultivar Septa, both higher and lower resistance to root hair infection was found in the accessions of the differentBrassica species. No complete resistance to root hair infection was found. Over the accessions studied, there was no correlation between the plant resistance estimated from greenhouse tests and the resistance to root hair infection of seedlings. The resistance of all accessions must at least partly be caused by other mechanisms which operate after the root hair plasmodia are formed.     

Indigenous highly virulent accessions of the sunflower root parasitic weed Orobanche cumana

Orobanche cumana (broomrape) is a severe constraint to sunflower production in southern and eastern Europe and the Middle East. Races A to E of this parasitic weed controlled by genes Or₁ to Or₅ have been described. A study of 38 seed accessions of O. cumana collected from different locations in Spain between 1983 and 2003 investigated the effect of long‐term storage in the laboratory on germination and infectivity and assessed their virulence on a number of sunflower cultivars. Only 18 infected the susceptible cultivar B117. Infectivity was maintained for up to 17 years of storage, but with a greatly decreased vigour as compared with that of recently collected seed. The 12 oldest viable accessions overcame the resistance of the gene Or₅ (in resistant line NR5). Seven out of them, in particular those collected in 1988 and 1989, were identified as race F. Three accessions were identified as race E allegedly holding components of higher virulence. Our results show evidence of the occurrence of race F prior to the use of sunflower hybrids resistant to race E, suggesting the former as indigenous to the country. This finding suggests the necessity of a continuous breeding of sunflower for resistance to O. cumana. The effectiveness and sustainability of genetic resistance must rely on the knowledge of the diverse virulence characteristics of O. cumana accessions.     

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.     

Domestication affected the basal and induced disease resistance in common bean (Phaseolus vulgaris)

Crop plants exhibit reduced levels of disease resistance, but little is known about the specific resistance mechanisms that are affected by breeding for increased yields. We investigated basal and chemically induced resistance of two wild accessions and four cultivars (including one landrace and three ??modern??, yield-improved cultivars that have been produced by hybridisation and pedigree breeding) of common bean (Phaseolus vulgaris) under greenhouse and field conditions. After treatment with benzothiadiazole, a widely used inducer of systemic acquired resistance, plants were challenged with one of two bacterial pathogens (Pseudomonas syringae pv. syringae and Enterobacter sp. strain FCB1). Basal resistance to Pseudomonas in the wild accessions was significantly higher than in the cultivars. Moreover, benzothiadiazole-treatment elevated resistance to the same pathogen in a wild accession and the landrace, but not in the yield-improved cultivars. Similarly, benzothiadiazole-induced resistance to Enterobacter FCB1 was detected in both wild accessions and the landrace, whereas the same treatment enhanced susceptibility to Enterobacter FCB1 in two of the yield-improved cultivars. Basal resistance to Pseudomonas was highly (but negatively) correlated to induced resistance over all accessions, and basal and inducible resistance to Enterobacter FCB1 were negatively correlated for the cultivars, but not when considering all six accessions. Benzothiadiazole-treatment increased growth rates under pathogen pressure of the wild accessions but not the cultivars. Apparently, the yield-improved cultivars investigated here have lost a considerable part of the basal and induced broad-spectrum disease resistance that characterises their wild relatives and to some degree also the landrace. Two of the yield-improved cultivars even became highly susceptible to infection by an Enterobacter strain that has not yet been described as a pathogen of bean and that is likely to represent a common environmental or phyllosphere bacterium. Future studies should disentangle the effects of domestication on the various layers of plant resistance to pathogens and consider the potential of wild accessions and landraces for future breeding programmes.     

Resistance to powdery mildew (Blumeria graminis f.sp. avenae) in oat seedlings and adult plants

In this work, 165 Avena sativa and Avena byzantina accessions were screened for resistance to powdery mildew caused by Blumeria graminis f.sp. avenae and the defence mechanisms of resistant plants were further characterized. Ten resistant and moderately resistant accessions were selected according to macroscopic assessment. A detailed histological study of selected genotypes showed a range of defence mechanisms, acting alone or in combination, that impeded fungal development at different stages. Since the resistance observed in the collection was scarce, a study of adult plant resistance was carried out in 45 genotypes selected from field trials. Nine oat landraces and two commercial varieties showed very high levels of adult plant resistance. A detailed study of the components of the adult plant resistance revealed a high increase of penetration and post‐haustorial resistance in the fifth compared to the first leaves. Identification of the resistance sources and characterization of underlying defence mechanisms will be useful for future breeding programmes and for further cellular and molecular studies to unravel the genetic basis of resistance, in this species in particular and in cereal–powdery mildew interactions in general.     

New sources of resistance to Meloidogyne incognita race 3 in wild cotton accessions and histological characterization of the defence mechanisms

Accessions of Gossypium spp., some of them never previously tested, were evaluated for resistance to a local isolate of Meloidogyne incognita race 3 in greenhouse experiments. Nematode infection was characterized based on the galling and egg mass indexes and the reproduction factors (RF). Root‐knot nematode reproduction among the newly tested accessions varied from highly susceptible (AS0188, AS0189) to moderately resistant (MT123 no. 3), and some accessions showed highly reduced nematode reproduction (CIR1343, CIR1348, Fai Mui). Histological observations of two resistant accessions (G. barbadense CIR1348 and G. hirsutum TX‐25, respectively) showed that resistance occurs through a two‐stage mechanism in the first accession and through a single‐stage mechanism in the second. Parasitism is blocked early after second‐stage juvenile (J2) penetration or during its initial tissue migration (CIR1348) and the development of later‐stage juveniles into female adults is suppressed at a later stage (TX‐25 and CIR1348). Fluorescence and bright light microscopy showed that root cells surrounding nematodes exhibit a hypersensitivity‐like reaction, with the accumulation of presumably phenolic compounds and the presence of necrotic cells that limit the development of nematodes and the formation of giant cells. Underdeveloped giant cells with degenerated cytoplasmic content were found in small numbers in CIR1348 and in large numbers in TX‐25, along with deformed nematodes. The full characterization of the defence mechanisms of novel sources of resistance to the root‐knot nematode in cotton constitutes a first step towards their use in crop improvement.     

Crenate broomrape control in pea by foliar application of benzothiadiazole (BTH)

Parasitic plants are becoming a severe constraint on major agricultural crops in Mediterranean and tropical countries and the efficacy of available means of control is minimal. The problem is particularly severe in field pea, which is very sensitive to standard glyphosate treatments and in which little resistance has been identified. Systemic Acquired Resistance (SAR) has proven to be effective as a tool for controlling plant pathogens, including fungi, bacteria and viruses, but only recently has this phenomenon started to be evaluated as a control strategy against parasitic weeds. The present studies were conducted to evaluate the potential of SAR activation for broomrape control in pea. The effect of salicylic acid, glutathione and benzothiadiazole (BTH) in three different application methods was studied. Foliar application of 0.6–1.0 mM BTH, in the form of Bion 50 (50% a.i.), reduced broomrape infection under controlled conditions (growth chamber and greenhouse) by limiting the success in attachment and retarding the development of established tubercles. http://www.phytoparasitica.org posting Dec. 1, 2003.     

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.     

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.     

Greenhouse and field screening of wild<Emphasis Type="Italic">Lycopersicon</Emphasis> germplasm for resistance to the whitefly<Emphasis Type="Italic">Bemisia argentifolii</Emphasis>

Thirty-two accessions of wild tomato (Lycopersicon spp.) germplasm were evaluated for resistance to the whiteflyBemisia argentifolii Bellows ⇐p; Perring in a greenhouse choice bioassay. Density data were recorded for the adaxial and abaxial leaf surfaces for (i) all life stages ofB. argentifolii and (ii) types I, IV, V and VI trichomes. Individual plant selections (33 from 22 wild tomato accessions) with high resistance were subsequently tested in the field to verify the resistance found in the greenhouse screening. Resistance was defined by the density of all life stages of the whitefly observed on the eight leaflets sampled at nodes 5 and 7. Only type IV trichomes had a consistent (but low) and significant negative correlation between trichome density and whitefly density for various life stages. The highest whitefly resistance was observed inLycopersicon pennellii accessions LA 716, LA 1340 and LA 2560. The most resistantL. hirsutum f.typicum accessions were LA 1777 and LA 1353. http://www.phytoparasitica.org posting Dec. 9, 2002.     

The potential for the Rapid Screening of Potato Cultivars (Solanum tuberosum) for Resistance to Powdery Scab (Spongospora subterranea) using a Laboratory Bioassay

Powdery scab of potato, once established in a field, is difficult to control because of the longevity of the resting spores (cystosori) of the causal organism, Spongospora subterranea f.sp. subterranea. Host resistance is likely to be the most efficient in a long-term control strategy for preventing build-up of field inoculum and spread of the disease. Resistance screening of potato cultivars is mostly done in laborious field trials where disease development is likely to be unpredictable. A bioassay with potato tissue cultured plantlets and cystosori as inoculum is described and was tested for its potential to screen potato cultivars at an early stage for their relative susceptibility to powdery scab by comparing the lab results with field data. With cystosori inoculum of Swiss origin, the laboratory test showed clear differences between the potato cultivars in the severity of zoosporangial root infection which correlated better with ranked tuber infection data, compared to root galling. There are apparent differences in the relative trends in susceptibility between roots and tubers of five selected cultivars when using naturally infested soil instead of prepared cystosori as inoculum in the lab bioassay. Furthermore, differences in the severity of zoosporangial root infection of two selected cultivars were found when cystosori from different countries where used as inoculum. A possible host genotype × pathogen interaction is discussed. The bioassay has the potential to screen and select for resistant material at an early breeding stage thus making field trials not unnecessary but more economical. It will allow the use of a standard set of pathogen collections and facilitate testing for inoculum virulence in infested soils.