Quantitative aspects of Orobanche crenata infestation in faba beans as affected by abiotic factors and parasite soil seedbank

The interactions between the root parasitic weed Orobanche crenata Forsk. and its host plant faba bean ( Vicia faba L.) were quantified under controlled and field conditions at ICARDA's Tel Hadya research station. In the field experiments conducted in 1993–94 and 1994–95 faba beans were sown on two dates, in plots with 0, 50, 200 and 600 O. crenata seeds kg^–1 soil, under both limited and sufficient moisture supply. The effects of temperature on the duration of the early developmental stages of O. crenata were investigated in a growth chamber. The extent of O. crenata infestation was closely related to the number of parasite seeds in the soil. The seed-density treatment with 600 seeds kg^–1 soil resulted in complete crop failure. Furthermore, O. crenata infestation was higher under sufficient than under limiting water supply conditions, irrespective of sowing date. Only in the moderately infested plots, did shifting of the planting time of faba bean result in a significant decrease in parasite dry weight and an increase in crop seed yield. The timing of germination, attachment and further developmental stages of O. crenata was not related to faba bean growth stage and was affected primarily by soil temperature. The duration of O. crenata developmental stages was estimated using the thermal time concept. The relationship between total number of parasite attachments at the harvest of the faba bean crop and O. crenata seed density was dependent on maximum faba bean root-length density measured by the start of pod-filling in each treatment combination of sowing date and moisture supply. The results are discussed with reference to implications for the development of a dynamic simulation model for the prediction of faba bean yield losses caused by O. crenata .     

Effect of plant density on grain yield, root–length density and Orobanche crenata infestation in two faba bean genotypes

The effect of host plant cultivar and sowing density on Orobanche crenata Forsk. L. infestation in Vicia faba was studied in a field experiment in north–west Syria. Two faba bean genotypes. ILB 1814 (Syrian Local Large) and 402/29/84 (new breeding line from Egypt), were planted at four sowing densities. Whereas ILB 1814 was severely affected by O. crenata , 402/29/84 proved to be highly resistant. Sowing density did not have a significant effect on O. crenata dry weight in either genotype. However, the number of O. crenata attachments in ILB 1814 was positively correlated with plant density. The resistance of faba bean genotype 402/29/84 to O. crenata is due to: (a) less plant vigour and root–length density; (b) necrosis of host cells preventing O. crenata attack either before or just after penetration into the host root, or developing a barrier in the host root after the formation of a small tubercle: (c) early flowering and pod setting.     

Seed bank and other demographic parameters of broomrape (Orobanche crenata Forsk.) populations in faba bean (Vicia faba L.)

A study on the population dynamics of broom-rape (Orobanche crenata Forskal) in faba bean (Viciafaba L.) was conducted in two locations for 8 years. The O. crenata seed bank increased every year in the 20-cm depth arable layer until it reached approximately 4 million (M) seeds m^?2. Seed viability ranged between 53% and 68%, approximately half of the seeds remaining dormant. About 3 × 10^?3% of the seed bank became attached to the root system of the faba beans. Only 9% of the attached broomrapes developed and emerged from the soil, possibly reflecting high levels of intraspecific competition. Maximum broomrape seed production for a population of 53 emerged broomrapes per m² was approximately 4 M seeds m^?2. About 43% of the seeds produced were not incorporated into the soil, most probably as a result of their degradation, decomposition or dispersion. Stock semencier et autres paramètres démographiques de I'orobanche crénelée (Orobanche crenata Forsk.) dans des cultures de féverolle (Vicia faba L.) Une étude de la dynamique de populations d'orobanche crénelée (Orobanche crenata Forsk.) dans des cultures de féverolle (Viciafaba L.) a été menée pendant 8 années sur deux sites. Le stock semencier de O. crenata a augmenteéchaque année dans la couche (20 cm) arable du sol jusqu'à atteindre 4 millions (M) de graines m^?2. La viabilité des graines était comprise entre 53 et 68%, approximativement la moitié d'entre elles présentaient une dormance. Environ 3 × 10^?3% du stock semencier adhérait au système racinaire de la féverolle. Seulement 9% des orobanches adhérantes se développaient et levaient, ce qui pourrait refléter une compétition intraspécifique importante. La production maxi-male de graines par une population de 53 orobanches levées au m²était d'environ 4 M graines m^?2. Environ 43% des graines produites n'étaient pas incorporées au sol, très prob-ablement en raison de leur dégradation, de leur décomposition ou de leur dispersion. Bodensamenbank und andere populationsde-mographische Parameter der Sommerwurz Orobanche crenata Forsk. in Bestanden der Acker-Bohne (Vicia faba L.) Bei einer Untersuchung der Populationsdy-namik der Sommerwurz Orobanche crenata Forsk. in Beständen der Acker-Bohne (Vicia faba L.) an 2 Orten über 8 Jahre hin nahm die Bodensamenbank in der Pflugzone von 20 cm Tiefe jedes Jahr zu, bis sie rund 4 Millionen Samen m^?2 erreichte. Die Lebensfähigkeit der Samen lag bei 53 bis 68%, etwa die Hälfte war dormant. Ungefähr 3 × 10^?3% kamen mit den Bohnenwurzeln in Berührung. Nur 9% davon entwickelten sich und wuchsen heran, worin vielleicht der hohe Grad intraspezifischer Konkurrenz zum Ausdruck kommt. Die höchste Zahl gebildeter Samen eines Sommer-wurzbestands von 53 Pflanzen m^?2 war rund 4 Millionen. Etwa 43% der gebildeten Samen gelangten nicht in den Boden, wahrscheinlich weil sie zerstört oder verweht wurden.     

Uptake and translocation of imazethapyr in peas as affected by parasitism of Orobanche crenata and herbicide application methods

[¹⁴C]-Imazethapyr was applied as a seed treatment and at plant pre-emergence and post-emergence to peas ( Pisum sativum L.) parasitized by Orobanche crenata Forsk. Herbicide uptake increased with time regardless of the application method. Uptake reached about 98%, 89%, 81% and 94% of the total herbicide applied for the seed coating, seed soaking, pre-emergence and post-emergence treatments respectively. Herbicide translocation within the host plants consistently differed between O. crenata -infected and non-infected plants. High levels of ¹⁴C activity were accumulated by parasitic plants from the host. In non-infected pea plants, pods were stronger sinks for imazethapyr than the other parts of the plant, regardless of the application method. The herbicide distribution in the pea plant: O. crenata complex showed the same pattern regardless of the application methods. However, accumulation of radioactivity in the parasite was lower with pre-emergence and post-emergence application than with the seed treatments. In addition, radioactivity concentration in O. crenata plants was slightly higher when [¹⁴C]-imazethapyr was applied to pea seeds by coating than by soaking.     

Variation Among and Within Populations of the Parasitic Weed Orobanche crenata from Spain and Israel Revealed by Inter Simple Sequence Repeat Markers

ABSTRACT The patterns of genetic variation among Orobanche crenata populations from Spain and Israel were studied using radiolabeled inter simple sequence repeat amplification products that were separated in sequencing polyacrylamide gels. The analysis of molecular variance indicated that most of the genetic diversity was attributable to differences among individuals within a population although significant divergences were found between regions. The Jaccard's similarity matrix was analyzed by unweighted pair-group method with arithmetic average and the resultant dendrogram clearly divided six populations by region, with the Spanish populations being more similar to each other than the Israeli populations. These results are consistent with the predominantly allogamous behavior of O. crenata and the extremely efficient dispersal of its seeds.     

Disease resistance in Vicia faba and Phaseolus vulgaris

Droplets of spore suspensions of each of four isolates ofBotrytis cinerea but not those of each of four isolates ofB. fabae proved to contain an antifungal compound 24 h after application on pod tissue ofVicia faba. Partial inhibition of germ-tube growth of three highly pathogenic isolates ofB. fabae was caused at 2.5 times the concentration of inhibitor needed to cause similar inhibition of each isolate ofB. cinerea and a weakly pathogenic isolate ofB. fabae. After extraction, concentration and chromatographic separation, 5–10 times more inhibitor was obtained from lesions in pods caused byB. cinerea than from those caused byB. fabae. However, the amounts of inhibitor extracted from whole leaves bearing either large lesions caused byB. fabae or small lesions caused byB. cinerea were almost the same. It is suggested that infection by either fungus induces inhibitor formation, but thatB. fabae metabolizes the inhibitor to an inactive form.No relation was found between amounts of an inhibitor produced in droplets of spore suspensions 24 h after application on pods of differential varieties ofPhaseolus vulgaris and the disease reactions caused by races ofColletotrichum lindemuthianum. Each race appeared to have a similar sensitivity to the inhibitor. Anatomical studies showed that only superficial growth of germ-tubes occurred in seed cavities in the first two days, after which penetration took place. Resistant or susceptible reactions were distinguished after 6 days in young pods, and even later in old pods. Before rejecting the hypothesis that the inhibitor may have a role in the mechanism of disease resistance, amounts of inhibitor in and around infection sites on leaves or stems should be measured. Apparent protection of leaf areas against infection was caused by prior inoculation with a race which was avirulent on the leaf. This phenomenon is consistent with the action of an inhibitor of the type found in pod tissues, but could be caused by reactions as yet unknown.     

Orobanche crenata (Forsk) control in Vicia faba (L.) with glyphosate as affected by herbicide rates and parasite growth stages

The broomrape (Orobanche crenata Forsk) susceptibility to glyphosate applied on faba-bean (Vicia faba L.) as affected by the parasite growth stages at the time of application was studied under field conditions. Glyphosate treatments delayed O. crenata emergence. Single glyphosate application to faba-bean infected with O. crenata predominantly at the stage (a) (small nodule) and (b) (nodules with initial vestigial roots) resulted in a moderate to low control. When the stages (c) (shoot bud already visible) or (d) (shoot and vestigial roots well developed) were the predominant stages, an excellent control was achieved with a single glyphosate application at 60 g ha^?1. Increased development stages, with the shoot emerged from the shoot bud, decreased its susceptibility to glyphosate. La lutte contre Orobanche crenata en culture de férerole avec le glyphosate; influence des doses d'herbicide et du stade de développement du parasite     

Gene expression profiling of Medicago truncatula roots in response to the parasitic plant Orobanche crenata

Orobanche crenata is a root parasitic weed that is a major constraint for grain and forage legume cultivation in Mediterranean and West Asia. Only moderate to low levels of incomplete resistance of complex inheritance has been identified so far in legume crops, which has hampered genetic and genomic analysis. In the present study, we provide a gene expression profile of roots of the model legume Medicago truncatula in response to infection by O. crenata . M. truncatula accessions SA27774 (complete resistance acting at early penetration stages) and SA4087 (incomplete late acting resistance mediated by necrosis of parasite tubercle) were inoculated with O. crenata seeds in a semi-sterile dish system. Roots were harvested at 15 (first contacts of the parasitism structures with the host roots), 21 (initial stage of parasite tubercle formation on SA4087) and 35 (prior necrosis of well-developed parasite tubercle of on SA4087) days post-inoculation. Array hybridisations revealed several hundred genes up-regulated in response to O. crenata infection. Gene expression patterns suggest that resistance mechanisms activated in both genotypes are temporally and spatially different and resemble those associated with plant resistance to microbial pathogens. Regulated genes identified here represent a comprehensive resource that can be used as a support to breeding strategies for resistance.     

Parasitic plant infection is partially controlled through symbiotic pathways

Legumes are unique in interacting with Rhizobium , arbuscular mycorrhizal (AM) fungi, and parasitic plants. To dissect common parts of these three plant–organism interactions, infection by Orobanche crenata was studied in mutants with altered symbiotic phenotypes of Medicago truncatula and Pisum sativum . Orobanche crenata inoculation of mutant lines carrying defective mutation in the genes dmi2 / sym19 and dmi3 resulted in an increase in O. crenata establishment. Similarly, inoculation of mutants carrying mutation in the gene sunn / sym29 that controls the autoregulation mechanism of the symbiosis, also lead to a significant increase in haustoria formation. Altogether, our results suggest that parasitic plant infection is partly controlled by both the conserved symbiotic pathway that mediates symbiont recognition and establishment and the autoregulation mechanism that regulates the extent of colonisation by Rhizobium and AM fungi.     

蚕豆抗锈病鉴定方法的改进及资源筛选

 蚕豆锈病是由蚕豆单胞锈菌 [Uromyces viciae-fabae (Pers.) Schroet ] 引起的蚕豆最重要的病害之一。为提高蚕豆锈病抗性种质筛选的效率及准确性,本文采用混合菌株在温室条件下苗期接种、成株期评价的方法进行抗性鉴定。以抗病材料K0772和感病材料K90315为对照,对1416份材料(3个年度鉴定材料数量分别为290、517和609份)进行鉴定试验,其中重复鉴定材料77份。抗病对照K0772历年均表现为抗病(R);感病对照K90315分别表现为感病(S)、感病(S)和高感(HS)。不同年份间的病情指数T检验结果显示,对照品种K0772和K90315及重复鉴定材料的3年重复鉴定结果间差异并不显著,具有较好的稳定性。同时,重复鉴定材料组间分别呈极显著、显著、极显著正相关。研究结果表明年度间的重复鉴定结果高度一致,具有较好的稳定性和重复性。通过3年重复鉴定,获得高抗(HR)材料12份,抗病(R)材料47份,中抗(MR)材料18份。因此,利用混合菌源进行苗期接种和在成株期对蚕豆品种锈病抗性进行评价的方法具有较好的稳定性和准确性,该方法可以用于蚕豆锈病综合抗性材料的筛选和抗病育种研究。     

Wheat/faba bean intercropping improves physiological and structural resistance of faba bean to fusaric acid stress

Fusarium oxysporum f. sp. fabae is the causal agent of fusarium wilt. Fusaric acid (FA), produced by F. oxysporum, plays an important role in the occurrence of disease, and intercropping is an effective measure for control of disease and for improving host resistance in plants. The objective of this study was to investigate the physiological and biochemical responses, and mechanisms of tissue structure resistance, of intercropped faba beans following exposure to different concentrations of FA. Results demonstrated that intercropping reduced the occurrence of fusarium wilt, and improved faba bean growth and yield. In addition, wheat intercropping significantly reduced red ink absorption of faba bean (33.2%), increased water content (3.1%), and increased activity of the root antioxidant enzymes peroxidase (POD) and catalase (CAT) (26.3% and 2.2.%, respectively). Furthermore, increased lignin content and callose deposition in plant vessels were observed (12.5% and 42.7%, respectively) when subjected to the highest concentration of FA stress (200 mg/L). Intercropping resulted in more intact root cell morphology, increased occurrence of intracellular vacuoles, increased cell wall thickness, and an increase in the number of mitochondria and rough endoplasmic reticulum. Intercropping alleviated the wilting effect of FA on faba bean via enhanced physiological, biochemical, and tissue structure resistance of faba bean root.     

蚕豆抗锈病鉴定方法的改进及资源筛选

 蚕豆锈病是由蚕豆单胞锈菌 [Uromyces viciae-fabae (Pers.) Schroet ] 引起的蚕豆最重要的病害之一。为提高蚕豆锈病抗性种质筛选的效率及准确性,本文采用混合菌株在温室条件下苗期接种、成株期评价的方法进行抗性鉴定。以抗病材料K0772和感病材料K90315为对照,对1416份材料(3个年度鉴定材料数量分别为290、517和609份)进行鉴定试验,其中重复鉴定材料77份。抗病对照K0772历年均表现为抗病(R);感病对照K90315分别表现为感病(S)、感病(S)和高感(HS)。不同年份间的病情指数T检验结果显示,对照品种K0772和K90315及重复鉴定材料的3年重复鉴定结果间差异并不显著,具有较好的稳定性。同时,重复鉴定材料组间分别呈极显著、显著、极显著正相关。研究结果表明年度间的重复鉴定结果高度一致,具有较好的稳定性和重复性。通过3年重复鉴定,获得高抗(HR)材料12份,抗病(R)材料47份,中抗(MR)材料18份。因此,利用混合菌源进行苗期接种和在成株期对蚕豆品种锈病抗性进行评价的方法具有较好的稳定性和准确性,该方法可以用于蚕豆锈病综合抗性材料的筛选和抗病育种研究。     

Effect of temperature and humidity on the longevity of Orobanche seeds

The effects of moisture and temperature on seed longevity were studied over periods of up to 426 days in three species of Orobanche . Saturated salt solutions were used to achieve equilibrium relative humidities (e.r.h.) between 11% and 85% at temperatures of 20–60 °C. The viability equation describing loss of viability in these conditions applied to the three species such that the lower the temperature and lower the equilibrium relative humidity the greater the longevity. The initial viability differed between species with the seed lot of Orobanche crenata having the lowest viability. However, the relative effect of temperature on longevity and the relative sensitivity of seed longevity to changes in equilibrium relative humidity were the same in all species. Longevity, estimated by the standard deviation of the seed survival curves was greater in Orobanche aegyptiaca and O. crenata than in Orobanche minor , being for example about 100, 103 and 49 days, respectively, at 40 °C, 50% e.r.h. Accurate prediction of longevity is necessary to understand and quantify the population dynamics of weeds and will help the farmer to optimize control measures such as solarization. Taken in conjunction with published data on loss of viability of imbibed seeds, the annual loss of viability on a typical Eritrean farm is predicted to be about 38%.     

Effects of phenylpropanoid and energetic metabolism inhibition on faba bean resistance mechanisms to rust

ABSTRACT Effects on penetration and hypersensitive resistance of the cinnamyl acid dehydrogenase (CAD) suicide inhibitor ([(2-hydroxyphenyl) amino] sulphinyl) acetic acid, 1.1 dimethyl ester, which suppresses phenylpro-panoid biosynthesis, and of D-mannose, which sequesters phosphate and reduces energy available in host cells, were studied in faba bean (Vicia faba) genotypes with differing resistance mechanisms to faba bean rust (Uromyces viciae-fabae). Inhibition of CAD reduced penetration resistance in lines 2N-34, 2N-52, V-1271, and V-1272, revealing an important role for phenylpropanoid biosynthesis in the resistance of these lines. Inhibition of CAD also inhibited hypersensitive cell death in these lines. D-mannose had little or no effect on resistance. By contrast, CAD inhibition did not affect penetration resistance of line BPL-261, which has a high degree of penetration resistance not associated with hypersensitive cell death. In BPL-261, D-mannose inhibited penetration resistance. The parallelism between the faba bean genotype responses to rust observed here and the response of barley genotypes with differing resistance mechanisms to powdery mildew after similar inhibitor treatments is analyzed and discussed.     

龟甲冬青枝枯病及其病原菌鉴定

于2002-2005年连续3年观察了杭州龟甲冬青枝枯病的症状,该病害先在嫩枝枝条发病,病斑初为水渍状斑点,后扩大成暗褐色的不规则形病斑。从发病部位分离培养出病原菌,根据其分生孢子的形态、大小和核糖体DNA内转录间隔区的序列,鉴定为褐纹拟茎点霉［Phomopsis vexans (Sacc.et Syd.) Harter］。本文对该病害症状和病原形态特征进行了描述。     

Induction of systemic resistance to rust in Vicia faba by phosphate and EDTA: effects of calcium

Resistance in the upper leaves of broad bean ( Vicia faba ) plants to infection by the rust fungus, Uromyces viciae-fabae, was increased following treatment of the lower leaves with 10 m m potassium phosphate or 5 m m EDTA. Increasing the interval between treatment of the lower leaves and inoculation of the upper leaves had little effect on rust infection. Thus rust infection was reduced by 50 and 34% if the upper leaves were inoculated 1 day after treatment of the lower leaves with potassium phosphate or EDTA, respectively, while there was a 75% reduction in infection if the interval between treatment and inoculation was increased to 12 days. Application of calcium nitrate (10 m m ) after the phosphate or EDTA treatments prevented the induction of systemic resistance. Calcium nitrate applied alone to the lower leaves had no significant effect on rust infection of the upper leaves.     

Characterization of new sources of resistance to Uromyces viciae-fabae in a germplasm collection of Vicia faba

A collection of 648 accessions of Vicia faba was screened for resistance to faba bean rust ( Uromyces viciae-fabae ). Two distinct types of resistance were identified, both resulting in reduced disease severity (DS) and area under the disease progress curve (AUDPC), but differing in the expression of hypersensitivity. One should be regarded as incomplete nonhypersensitive resistance and the other as incomplete resistance with late hypersensitivity. The hypersensitive resistance, which has not been reported before, was not dependent on temperature or plant age. These two types of resistance were characterized by three macroscopic components of resistance: increased latent period (LP), decreased colony size (CS) and a relatively reduced infection frequency (IF), both on seedlings and on adult plants. LP and CS were the components of nonhypersensitive resistance most highly correlated with DS and AUDPC measured under field conditions. The presence of necrosis was an additional component in the hypersensitive resistant response.     

Screening faba bean for chocolate spot resistance: evaluation methods and effects of age of host tissue and temperature

Chocolate spot is an important disease of faba bean (Vicia faba) caused by the necrotrophic fungus Botrytis fabae. The aims of this work were: i) to compare different methods of screening for resistance; ii) to assess the influence of the age of host tissue and temperature on this pathosystem. To this effect, a collection of 42 faba bean accessions was evaluated in mature plant stage in the field in Cordoba (Southern Spain) and in detached leaflet and whole plant tests under controlled conditions. Field results correlated better with those of the whole plant test than with those of the detached leaflet assay. Integration of results from the field and whole plant experiments resulted in the selection of six accessions of interest as sources of resistance. Influence of leaf age on disease development was found to be genotype dependent. Older leaves were more susceptible than younger ones in 23 accessions, while no difference between leaf ages was detected in the remaining accessions. The effects of plant age and temperature were assessed by a whole plant test on seven accessions at two plant ages (4 and 7 weeks) and three temperatures (13, 20, and 25°C). Results showed that the differential genotypic responses to B. fabae were not significantly influenced by either plant age or temperature, although there was a tendency towards lower susceptibility to chocolate spot in faba bean plants as they become older. Further, a partial high-temperature, young-plant resistance was detected.