Phelipanche Aegyptiaca Research Articles

A modelling approach for predicting the initial phase of Egyptian broomrape (Phelipanche aegyptiaca) parasitism in potato

Phelipanche aegyptiaca is one of the most important parasitic weeds in some crops including potato. P.aegyptiaca reduces potato tuber size and can produce severe yield losses. The development of predictive models can be useful to help managers to choose the best management options and times and, thereby, improve weed control.The objective of this study was to develop a predictive thermal time model of the P.aegyptiaca establishment in potato using field experiment data. The relationship between the cumulative attachments of P.aegyptiaca and air/soil thermal time was analysed using Gompertz and Weibull functions. The Weibull soil thermal time model produced the better fit and was the more plausible one. The latter model was successfully validated under field conditions and can be used as a predictive tool contributing to optimize the timing of P.aegyptiaca control. According to the weibull model and soil thermal time the lag time and 50% of P.aegyptiaca attachments occurred in 613.75 (124.8) and 999.49 (5.98) TT respectively.

Spatial Spread of the Root Parasitic Weed Phelipanche aegyptiaca in Processing Tomatoes by Using Ecoinformatics and Spatial Analysis.

Egyptian broomrape (Phelipanche aegyptiaca) is one of the main threats to tomato production in Israel. The seed bank of P. aegyptiaca rapidly develops and spreads in the field. Knowledge about the spatio-temporal distribution of such weeds is required in advance of emergence, as they emerge late in their life cycle when they have already caused major crop damage. The aim of this study is to reveal the effects of two major internal infestation sources: crop rotation and infestation history; and one external source: proximity to infested tomato fields; on infestation of P. aegyptiaca in processing tomatoes. Ecoinformatics, spatial analysis and geostatistics were used to examine these effects. A regional survey was conducted to collect data on field history from 238 tomato fields between 2000 and 2012, in a major tomato-growing region in Israel. Multivariate logistic regression in the framework of generalized linear models (GLM) has demonstrated the importance of all three variables in predicting infestation in tomato fields. The parameters of the overall model indicated a high specificity between tomatoes and P. aegyptiaca, which is potentially responsible for aggravating infestation. In addition, P. aegyptiaca infestation levels were intensively mapped in 43 of the 238 tomato fields in the years 2010–2012. Geostatistical measures showed that 40% of the fields had clustered infestation spatial patterns with infestation clusters located along the fields’ borders. Strong linear and negative relationships were found between infestation level and distance from a neighboring infested field, strengthening the role of infested tomato fields in P. aegyptiaca spread. An experiment specifically designed for this study showed that during harvest, P. aegyptiaca seeds are blown from an infested field to a distance of at least 90 m, and may initiate infestation in neighboring fields. Integrating current knowledge about the role of agricultural practices on the spread of P. aegyptiaca with the results of this study enabled us to propose a mechanism for the spread of P. aegyptiaca. Given the major effect of agricultural practices on infestation levels, it is assumed that the spread of this weed can be suppressed by implementing sanitation and using decision support tools for herbicide application.

Non-chemical Control of Root Parasitic Weeds with Biochar.

This study tested whether soil-applied biochar can impact the seed germination and attachment of root parasitic weeds. Three hypotheses were evaluated: (i) biochar adsorbs host-exuded signaling molecules; (ii) biochar activates plants’ innate system-wide defenses against invasion by the parasite; and (iii) biochar has a systemic influence on the amount of seed germination stimulant produced or released by the host plant. Three types of experiments were performed: (I) pot trials with tomato (Solanum lycopersicum) infested with Phelipanche aegyptiaca PERS. (Egyptian broomrape) and three different types of biochar at concentrations ranging from 0 to 1.5% weight, wherein tomato plant biomass, P. aegyptiaca biomass, and number of P. aegyptiaca-tomato root attachments were quantified; (II) split-root biochar/no-biochar experiments under hydroponic growing conditions performed in polyethylene bags with tomato plant rootings, wherein P. aegyptiaca seed germination percentage and radicle attachment numbers were quantified; and (III) germination trials, wherein the effect of biochar adsorption of GR-24 (artificial germination stimulant) on P. aegyptiaca seed germination was quantified. Addition of biochar to the pot soil (Experiment I) resulted in lower levels of P. aegyptiaca infection in the tomato plants, mainly through a decrease in the number of P. aegyptiaca attachments. This led to improved tomato plant growth. In Experiment II, P. aegyptiaca seed germination percentage decreased in the biochar-treated root zone as compared with the no-biochar control root zone; P. aegyptiaca radicle attachment numbers decreased accordingly. This experiment showed that biochar did not induce a systemic change in the activity of the stimulant molecules exuded by the tomato roots, toxicity to the radicles, or a change in the ability of the radicles to penetrate the tomato roots. The major cause for the decrease in germination percentage was physical adsorption of the stimulant molecule by the biochar (Experiment III). Adding biochar to soil to reduce infections by root parasitic weeds is an innovative means of control with the potential to become an important strategy both for non-chemical treatment of this family of pests, and for enhancing the economic feasibility of the pyrolysis/biochar platform. This platform is often viewed as one of a handful of credible strategies for helping to mitigate climate change.

The Influence of the Plant Growth Regulator Maleic Hydrazide on Egyptian Broomrape Early Developmental Stages and Its Control Efficacy in Tomato under Greenhouse and Field Conditions.

Broomrapes (Phelipanche spp. and Orobanche spp.) are holoparasitic plants that cause tremendous losses of agricultural crops worldwide. Broomrape control is extremely difficult and only amino acid biosynthesis-inhibiting herbicides present an acceptable control level. It is expected that broomrape resistance to these herbicides is not long in coming. Our objective was to develop a broomrape control system in tomato (Solanum lycopersicum L.) based on the plant growth regulator maleic hydrazide (MH). Petri-dish and polyethylene-bag system experiments revealed that MH has a slight inhibitory effect on Phelipanche aegyptiaca seed germination but is a potent inhibitor of the first stages of parasitism, namely attachment and the tubercle stage. MH phytotoxicity toward tomato and its P. aegyptiaca-control efficacy were tested in greenhouse experiments. MH was applied at 25, 50, 75, 150, 300, and 600 g a.i. ha-1 to tomato foliage grown in P. aegyptiaca-infested soil at 200 growing degree days (GDD) and again at 400 GDD. The treatments had no influence on tomato foliage or root dry weight. The total number of P. aegyptiaca attachments counted on the roots of the treated plants was significantly lower at 75 g a.i. ha-1 and also at higher MH rates. Phelipanche aegyptiaca biomass was close to zero at rates of 150, 300, and 600 g a.i. ha-1 MH. Field experiments were conducted to optimize the rate, timing and number of MH applications. Two application sequences gave superior results, both with five split applications applied at 100, 200, 400, 700, and 1000 GDD: (a) constant rate of 400 g a.i. ha-1; (b) first two applications at 270 g a.i. ha-1 and the next three applications at 540 g a.i. ha-1. Based on the results of this study, MH was registered for use in Israel in 2013 with the specified protocol and today, it is widely used by most Israeli tomato growers.

The Effects of Herbicides Targeting Aromatic and Branched Chain Amino Acid Biosynthesis Support the Presence of Functional Pathways in Broomrape.

It is not clear why herbicides targeting aromatic and branched-chain amino acid biosynthesis successfully control broomrapes—obligate parasitic plants that obtain all of their nutritional requirements, including amino acids, from the host. Our objective was to reveal the mode of action of imazapic and glyphosate in controlling the broomrape Phelipanche aegyptiaca and clarify if this obligatory parasite has its own machinery for the amino acids biosynthesis. P. aegyptiaca callus was studied to exclude the indirect influence of the herbicides on the parasite through the host plant. Using HRT – tomato plants resistant to imidazolinone herbicides, it was shown that imazapic is translocated from the foliage of treated plants to broomrape attachments on its roots and controls the parasite. Both herbicides inhibited P. aegyptiaca callus growth and altered the free amino acid content. Blasting of Arabidopsis thaliana 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) and acetolactate synthase (ALS) cDNA against the genomic DNA of P. aegyptiaca yielded a single copy of each homolog in the latter, with about 78 and 75% similarity, respectively, to A. thaliana counterparts at the protein level. We also show for the first time that both EPSPS and ALS are active in P. aegyptiaca callus and flowering shoots and are inhibited by glyphosate and imazapic, respectively. Thus leading to deficiency of those amino acids in the parasite tissues and ultimately, death of the parasite, indicating the ability of P. aegyptiaca to synthesize branched-chain and aromatic amino acids through the activity of ALS and EPSPS, respectively.

Metabolic Investigation of Phelipanche aegyptiaca Reveals Significant Changes during Developmental Stages and in Its Different Organs.

Phelipanche aegyptiaca Pers. is a root holoparasitic plant considered to be among the most destructive agricultural weeds worldwide. In order to gain more knowledge about the metabolic profile of the parasite during its developmental stages, we carried out primary metabolic and lipid profiling using GC-MS analysis. In addition, the levels of amino acids that incorporate into proteins, total protein in the albumin fraction, nitrogen, reduced sugars, and phenols were determined. For the assays, the whole plants from the four developmental stages—tubercle, pre-emergent shoot, post-emergent shoot, and mature flowering plants—were taken. Thirty-five metabolites out of 66 differed significantly between the various developmental stages. The results have shown that the first three developmental stages were distinguished in their profiles, but the latter two did not differ from the mature stage. Yet, 46% of the metabolites detected did not change significantly during the developmental stages. This is unlike other studies of non-parasitic plants showing that their metabolic levels tend to alter significantly during development. This implies that the parasite can control the levels of these metabolites. We further studied the metabolic nature of five organs (adventitious roots, lower and upper shoot, floral buds, and flowers) in mature plants. Similar to non-parasitic plants, the parasite exhibited significant differences between the vegetative and reproductive organs. Compared to other organs, floral buds had higher levels of free amino acids and total nitrogen, whereas flowers accumulated higher levels of simple sugars such as sucrose, and the putative precursors for nectar synthesis, color, and volatiles. This suggests that the reproductive organs have the ability to accumulate metabolites that are required for the production of seeds and as a source of energy for the reproductive processes. The data contribute to our knowledge about the metabolic behavior of parasites that rely on their host for its basic nutrients.

Host-Parasite-Bacteria Triangle: The Microbiome of the Parasitic Weed Phelipanche aegyptiaca and Tomato-Solanum lycopersicum (Mill.) as a Host.

Broomrapes (Phelipanche/Orobanche spp.) are holoparasitic plants that subsist on the roots of a variety of agricultural crops, establishing direct connections with the host vascular system. This connection allows for the exchange of various substances and a possible exchange of endophytic microorganisms that inhabit the internal tissues of both plants. To shed some light on bacterial interactions occurring between the parasitic Phelipanche aegyptiaca and its host tomato, we characterized the endophytic composition in the parasite during the parasitization process and ascertained if these changes were accompanied by changes to endophytes in the host root. Endophyte communities of the parasitic weed were significantly different from that of the non-parasitized tomato root but no significant differences were observed between the parasite and its host after parasitization, suggesting the occurrence of bacterial exchange between these two plants. Moreover, the P. aegyptiaca endophytic community composition showed a clear shift from gram negative to gram-positive bacteria at different developmental stages of the parasite life cycle. To examine possible functions of the endophytic bacteria in both the host and the parasite plants, a number of unique bacterial candidates were isolated and characterized. Results showed that a Pseudomonas strain PhelS10, originating from the tomato roots, suppressed approximately 80% of P. aegyptiaca seed germination and significantly reduced P. aegyptiaca parasitism. The information gleaned in the present study regarding the endophytic microbial communities in this unique ecological system of two plants connected by their vascular system, highlights the potential of exploiting alternative environmentally friendly approaches for parasitic weed control.

Secondary Effects of Glyphosate Action in Phelipanche aegyptiaca: Inhibition of Solute Transport from the Host Plant to the Parasite

It is currently held that glyphosate efficiently controls the obligate holoparasite Phelipanche aegyptiaca (Egyptian broomrape) by inhibiting its endogenous shikimate pathway, thereby causing a deficiency in aromatic amino acids (AAA). While there is no argument regarding the shikimate pathway being the primary site of the herbicide's action, the fact that the parasite receives a constant supply of nutrients, including proteins and amino acids, from the host does not fit with an AAA deficiency. This apparent contradiction implies that glyphosate mechanism of action in P. aegyptiaca is probably more complex and does not end with the inhibition of the AAA biosynthetic pathway alone. A possible explanation would lie in a limitation of the translocation of solutes from the host as a secondary effect. We examined the following hypotheses: (a) glyphosate does not affects P. aegyptiaca during its independent phase and (b) glyphosate has a secondary effect on the ability of P. aegyptiaca to attract nutrients, limiting the translocation to the parasite. By using a glyphosate-resistant host plant expressing the “phloem-mobile” green fluorescent protein (GFP), it was shown that glyphosate interacts specifically with P. aegyptiaca, initiating a deceleration of GFP translocation to the parasite within 24 h of treatment. Additionally, changes in the entire sugars profile (together with that of other metabolites) of P. aegyptiaca were induced by glyphosate. In addition, glyphosate did not impair germination or seedling development of P. aegyptiaca but begun to exert its action only after the parasite has established a connection to the host vascular system and became exposed to the herbicide. Our findings thus indicate that glyphosate does indeed have a secondary effect in P. aegyptiaca, probably as a consequence of its primary target inhibition—via inhibition of the translocation of phloem-mobile solutes to the parasite, as was simulated by the mobile GFP. The observed disruption in the metabolism of major sugars that are abundant in P. aegyptiaca within 48 h after glyphosate treatment provides a possible explanation for this inhibition of translocation and might reflect a critical secondary effect of the herbicide's primary action that results in loss of the parasite's superior sink for solutes.

Molecular Identification of Broomrape Species from a Single Seed by High Resolution Melting Analysis.

Broomrapes are holoparasitic plants spreading through seeds. Each plant produces hundreds of thousands of seeds which remain viable in the soils for decades. To limit their spread, drastic measures are being taken and the contamination of a commercial seed lot by a single broomrape seed can lead to its rejection. Considering that broomrapes species identification from a single seed is extremely difficult even for trained botanists and that among all the described species, only a few are really noxious for the crops, numerous seed lots are rejected because of the contamination by seeds of non-noxious broomrape species. The aim of this study was to develop and evaluate a High Resolution Melting assay identifying the eight most noxious and common broomrape species (Phelipanche aegyptiaca, Orobanche cernua, O. crenata, O. cumana, O. foetida, O. hederae, O. minor, and P. ramosa) from a single seed. Based on trnL and rbcL plastidial genes amplification, the designed assay successfully identifies O. cumana, O. cernua, O. crenata, O. minor, O. hederae, and O. foetida; P. ramosa, and P. aegyptiaca can be differentiated from other species but not from each other. Tested on 50 seed lots, obtained results perfectly matched identifications performed by sequencing. Through the analysis of common seed lots by different analysts, the reproducibility of the assay was evaluated at 90%. Despite an original sample preparation process it was not possible to extract enough DNA from some seeds (10% of the samples). The described assay fulfills its objectives and allows an accurate identification of the targeted broomrape species. It can be used to identify contaminants in commercial seed lots or for any other purpose. The assay might be extended to vegetative material.

Thermal Time Model for Egyptian Broomrape (Phelipanche aegyptiaca) Parasitism Dynamics in Carrot (Daucus carota L.): Field Validation.

Carrot, a highly profitable crop in Israel, is severely damaged by Phelipanche aegyptiaca parasitism. Herbicides can effectively control the parasite and prevent damage, but for optimal results, knowledge about the soil–subsurface phenological stage of the parasite is essential. Parasitism dynamics models have been successfully developed for the parasites P. aegyptiaca, Orobanche cumana, and Orobanche minor in the summer crops, tomato, sunflower, and red clover, respectively. However, these models, which are based on a linear relationship between thermal time and the parasitism dynamics, may not necessarily be directly applicable to the P. aegyptiaca–carrot system. The objective of the current study was to develop a thermal time model to predict the effect of P. aegyptiaca parasitism dynamics on carrot growth. For development and validation of the models, data was collected from a temperature-controlled growth experiment and from 13 plots naturally infested with P. aegyptiaca in commercial carrot fields. Our results revealed that P. aegyptiaca development is related to soil temperature. Moreover, unlike P. aegyptiaca parasitism in sunflower and tomato, which could be predicted both a linear model, P. aegyptiaca parasitism dynamics on carrot roots required a nonlinear model, due to the wider range of growth temperatures of both the carrot and the parasite. Hence, two different nonlinear models were developed for optimizing the prediction of P. aegyptiaca parasitism dynamics. Both models, a beta function model and combined model composed of a beta function and a sigmoid curve, were able to predict first P. aegyptiaca attachment. However, overall P. aegyptiaca dynamics was described more accurately by the combined model (RMSE = 14.58 and 10.79, respectively). The results of this study will complement previous studies on P. aegyptiaca management by herbicides to facilitate optimal carrot growth and handling in fields infested with P. aegyptiaca.

Characterization of Resistance Mechanisms in Faba Bean (Vicia faba) against Broomrape Species (Orobanche and Phelipanche spp.).

Faba bean (Vicia faba) production in Mediterranean and Near East agriculture is severely constrained by broomrape infection. The most widely distributed broomrape species affecting faba bean is Orobanche crenata, although O. foetida and Phelipanche aegyptiaca are of local importance. Only moderately resistant cultivars are available to farmers. Rizotrons studies allowed the dissection of resistance components in faba bean accessions against the very infective species O. crenata, O. foetida var. broteri and P. aegyptiaca, and to the inappropriate P. ramosa and O. foetida var. foetida. Results confirm that some levels of incomplete resistance are available, resulting in a reduced number of broomrape tubercles successfully formed per faba bean plant. Interestingly, the intermediate levels of resistance of cv. Baraca were operative against all broomrape populations and species studied, confirming previous reports on the stability of resistance of Baraca in field trials in different countries. Low induction of seed germination played a major role in the resistance against the inappropriate O. foetida var. foetida but not against the also inappropriate P. ramosa, neither to the infective species O. crenata, O. foetida var. broteri, or P. aegyptiaca. Negative tropism of germinated seeds with radicles growing away from faba bean roots was marked for both inappropriate species but was not observed in any of the infective species. Also, a proportion of radicles that had successfully contacted faba bean roots became necrotic, failing in starting tubercle development, particularly frequent for the two inappropriate species. Such necrosis was significant also on radicles contacting resistant faba bean accessions, being particularly relevant for Spanish O. crenata population, and lower although still significant in some accessions against Syrian O. crenata and P. aegyptiaca, suggesting that this might also be an operative mechanism to be selected and further exploited in faba bean resistance breeding. Even formed broomrape tubercles might later become necrotic, particularly in the case of some of the resistant faba bean accessions to the Spanish O. crenata and to P. aegyptiaca but not to the very infective Syrian O. crenata or O. foetida var. broteri.

Primary metabolic profiling of Egyptian broomrape (Phelipanche aegyptiaca) compared to its host tomato roots

Broomrape (Phelipanche aegyptiaca) is a root holoparasitic plant considered among the most destructive agricultural weeds worldwide. In order to acquire more knowledge about the metabolism of broomrape and its interaction with its tomato host, we performed primary metabolic profiling using GCMS analysis for the early developmental stage of the parasite and of infected and non-infected roots. The analysis revealed that out of 59 metabolites detected, the levels of 37 significantly increased in the parasite while the levels of 10 significantly decreased compared to the infected roots. In addition, the analysis showed that the levels of total protein in the albumin fraction, reducing sugars (representing starch) and total phenols increased by 9.8-, 4.6- and 3.3-fold, respectively, in the parasite compared to the roots. These changes suggest that P. aegyptiaca has its own metabolism that differs significantly in its regulation from those found in their host. In addition, the results have shown that the levels of most of the metabolites in the infected roots were similar to levels detected in the non-infected roots, except for seven metabolites whose levels increased in the infected versus the non-infected roots. This suggests that the parasite did not significantly affect the host primary metabolic pathways.

Two hAT transposon genes were transferred from Brassicaceae to broomrapes and are actively expressed in some recipients.

A growing body of evidence is pointing to an important role of horizontal gene transfer (HGT) in the evolution of higher plants. However, reports of HGTs of transposable elements (TEs) in plants are still scarce, and only one case is known of a class II transposon horizontally transferred between grasses. To investigate possible TE transfers in dicots, we performed transcriptome screening in the obligate root parasite Phelipanche aegyptiaca (Orobanchaceae), data-mining in the draft genome assemblies of four other Orobanchaceae, gene cloning, gene annotation in species with genomic information, and a molecular phylogenetic analysis. We discovered that the broomrape genera Phelipanche and Orobanche acquired two related nuclear genes (christened BO transposase genes), a new group of the hAT superfamily of class II transposons, from Asian Sisymbrieae or a closely related tribe of Brassicaceae, by HGT. The collinearity of the flanking genes, lack of a classic border structure, and low expression levels suggest that BO transposase genes cannot transpose in Brassicaceae, whereas they are highly expressed in P. aegyptiaca.

Mechanism of glyphosate control of Phelipanche aegyptiaca.

Despite its total reliance on its host plant, the holoparasite Phelipanche aegyptiaca suffers from a deficiency of aromatic amino acids upon exposure to glyphosate. The herbicide glyphosate inhibits 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), a key enzyme in the biosynthesis of aromatic amino acids. However, the functionality of the EPSPS pathway in the obligate root holoparasite Phelipanche aegyptiaca is not straightforward because of the parasite's total dependence on the host plant. Despite the importance of glyphosate as a means of controlling P. aegyptiaca, the mechanism of action of the herbicide in this parasite is not clearly understood. We characterized glyphosate control of P. aegyptiaca by using a glyphosate-resistant tomato (GRT) genotype as the host plant and evaluating the activity of EPSPS and the levels of free aromatic amino acids in the parasite. The viability of the parasite's tissues deteriorated within the first 40h after treatment (HAT) with glyphosate. In parallel, shikimate accumulation in the parasite was first detected at 24 HAT and increased over time. However, shikimate levels in the GRT host did not increase, indicating that the host was indeed glyphosate tolerant. Free phenylalanine and tyrosine levels decreased by 48 HAT in the parasite, indicating a deficiency of aromatic amino acids. The use of GRT as the host enabled us to observe, in an in situ experimental system, both endogenous EPSPS inhibition and a deficiency of aromatic amino acids in the parasite. We thus provided evidence for the presence of an active EPSPS and aromatic amino acid biosynthesis pathway in P. aegyptiaca and pinpointed this pathway as the target of glyphosate action in this parasite.

Conditioning duration and agents involved in broomrape seeds responding to germination stimulants

Weedy broomrape species, such as sunflower broomrape (Orobanche cumana Wallr.) and Egyptian broomrape [Phelipanche aegyptiaca Pers. (syn. O. aegyptiaca)], require a period of pre-conditioning before they can respond to germination stimulants. Thus, the sensitivity of weedy broomrape seeds to germination stimulants could be an important factor for broomrape control. In this study, the influence of conditioning agents, conditioning period (0–21 days) and germination stimulants on the germination of sunflower broomrape and Egyptian broomrape seeds was analyzed. Without conditioning, the sunflower and Egyptian broomrape seeds exhibited negligible germination responses to the stimulants. The germination rate of the broomrape seeds increased rapidly with conditioning period and reached a maximum under a conditioning period of 4–10 days; further prolonged conditioning resulted in a decrease in the germination rate. Gibberellic acid (GA3) could not only break the dormancy of the sunflower and Egyptian broomrape seeds but also maintained the high sensitivity of these seeds even after 21 days of conditioning. Furthermore, 100 µM of GA3 induced the germination of the Egyptian broomrape seeds. The stimulants that induced Egyptian broomrape germination were ranked in decreasing order as GR24 (76.8 %), strigol (76.1 %), tobacco root exudates (49.5 %), dehydrocostus lactones (DCL, 39.2 %), and maize root exudates (18 %). In contrast, GA3 did not directly induce sunflower broomrape seed germination, which responded to strigol (62.8 %) > maize root exudates (58.2 %) > GR24 (57.9 %) > tobacco root exudates (41.6 %) > DCL (41.3 %). These results indicate specialized recognition of germination stimulants by sunflower and Egyptian broomrape. This study may contribute to a better understanding of parasitic weed germination and may lead to improved control strategies.

Global Transcriptomic Analysis Reveals the Mechanism of Phelipanche aegyptiaca Seed Germination.

Phelipanche aegyptiaca is one of the most destructive root parasitic plants of Orobanchaceae. This plant has significant impacts on crop yields worldwide. Conditioned and host root stimulants, in particular, strigolactones, are needed for unique seed germination. However, no extensive study on this phenomenon has been conducted because of insufficient genomic information. Deep RNA sequencing, including de novo assembly and functional annotation was performed on P. aegyptiaca germinating seeds. The assembled transcriptome was used to analyze transcriptional dynamics during seed germination. Key gene categories involved were identified. A total of 274,964 transcripts were determined, and 53,921 unigenes were annotated according to the NR, GO, COG, KOG, and KEGG databases. Overall, 5324 differentially expressed genes among dormant, conditioned, and GR24-treated seeds were identified. GO and KEGG enrichment analyses demonstrated numerous DEGs related to DNA, RNA, and protein repair and biosynthesis, as well as carbohydrate and energy metabolism. Moreover, ABA and ethylene were found to play important roles in this process. GR24 application resulted in dramatic changes in ABA and ethylene-associated genes. Fluridone, a carotenoid biosynthesis inhibitor, alone could induce P. aegyptiaca seed germination. In addition, conditioning was probably not the indispensable stage for P. aegyptiaca, because the transcript level variation of MAX2 and KAI2 genes (relate to strigolactone signaling) was not up-regulated by conditioning treatment.

Sesame genotype influences growth and phenology ofPhelipanche aegyptiaca

Egyptian broomrape (Phelipanche aegyptiaca) is a parasitic weed causing major yield loss in many field and vegetable crops. Recent observations in Iran suggest that sesame (Sesamum indicum) may serve either as a trap crop or as a host for broomrape, depending on genotype. The central objective of this research was to identify sesame genotypes that could act either as potential trap crops, or sources of genetic material for development of broomrape resistant cultivars. Seeds of 11 Iranian sesame genotypes (Ardestan, Esfarayen, Oltan, Ahvaz, Borazjan, Darab-14, Ferdous, Kalat, Naz Tak Shakhe, Varamin and Yekta) and 10 foreign genotypes (S16, S21, S23, S25, S4, S5, S6, S7, S8 and S9) were planted with broomrape seeds in pots. At the flowering stage of sesame, tubercle number and dry weight, shoot number and dry weight and shoot height of broomrape were determined. Shoot dry weight, root dry weight and seed yield of sesame were determined at the physiological maturity. The results showed that broomrape growth varied with sesame genotype. Egyptian broomrape dry weight measured from 1.6% (S23) to 35.4% (Kalat) of the dry weight of the sesame on which it grew. Broomrape total dry weight had positive and highly correlation with the root dry weight of the sesame genotype on which it grew. When broomrape planted with the S7, S8, S9, S21 and S23 genotypes produced tubercles, but no shoots on these genotypes. Such genotypes could be used as both trap crops and as breeding-sources for development of resistant sesame genotypes.

A large-scale analysis of soil and bioclimatic factors affecting the infestation level of tobacco (Nicotiana tabacum L.) by Phelipanche species

Soil and bioclimatic factors constitute key factors that influence the interaction between host plants and broomrape. The identification and quantification of the relationship between environmental variables and the level of infestation with Phelipanche aegyptiaca and Phelipanche ramosa was examined for 144 tobacco fields in surveys during two years 2003–2004 in Greece. The distribution of Phelipanche species was mapped using a Geographical Information System to reveal variability in the infestation level between the species, the cultivated types and the sampling areas. To assess the effect of abiotic factors on the level of infestation with Phelipanche species, soil and bioclimatic parameters were taken into account: soil texture, pH, organic matter content (OM), total humidity index (THI) and growing degree days (GDD). Univariate and multivariate statistics were used to examine the influence of the above parameters on Phelipanche intensity. ANOVA showed that the infestation varied significantly between the sampling areas and Phelipanche species, while significant differences were also detected among the regions in terms of the studied abiotic parameters. Correlation analysis demonstrated that the level of Phelipanche infestation correlated negatively with pH, THI and positively with OM. Multiple regression analysis suggested that soil and climatic variables together explained 63% of the infestation variance. Discriminant functions accounted for 90% of the total variation in the dataset. All statistical analyses demonstrated that pH, THI and OM were the most decisive variables for the severity of infestation. The two Phelipanche species were clearly discriminated, with P. ramosa being found at higher and P. aegyptiaca at a lower level of infestation. A parasitism specification was also observed between the two Phelipanche species and tobacco types. The findings provide constructive information for modelling broomrape infestation based on abiotic factors and create a baseline for future monitoring of broomrape distribution.

Effectiveness of ten commercial maize cultivars in inducing Egyptian broomrape germination

Egyptian broomrape (EB), Phelipanche aegyptiaca, is a devastating root parasite, causing enormous crop losses around the world. Maize has the potential to influence the growth of other plants through releasing certain allelochemicals and is able to induce germination of at least three broomrape species. To determine whether maize could be used as a trap crop for EB, 10 maize cultivars were tested for their ability to induce EB germination. The results showed that maize cultivars can induce EB germination, and that germination rates in a cut-root experiment and a hydroponic experi- ment were consistent. Maize cvs Changcheng 799 and Zhengdan 958 induced the highest EB germination rates, while cvs Luyu 13 and Zhengyu 203 were the least effective. These four maize cultivars were further studied in a pot experiment. Rhizosphere soil, rhizosphere soil extracts, root extracts and shoot extracts from these cultivars were all able to induce EB germination, with cv. Changcheng 799 inducing the highest germination rates. Root extracts generally induced higher germination rates than shoot extracts. It is suggested that Changcheng 799 could be planted as a trap crop for control of EB.

Germination of Phelipanche aegyptiaca and Cuscuta campestris seeds in composted farm manure

The seed vitality of the parasitic weeds Phelipanche aegyptiaca (Egyptian broomrape) and Cuscuta campestris (field dodder) is a highly important factor in their dissemination to nearby and distant habitats. Fresh manure composting is an essential process as a ‘good agricultural practice’ of using manure safely as a fertilizer without dissemination of plant diseases, propagules or weed seeds. The aims of this research were a) to determine whether composting is effective in eliminating P. aegyptiaca and C. campestris seed vitality b) to evaluate the temperatures needed to eliminate seed viability of these species. Seeds of P. aegyptiaca and C. campestris were placed in various depths of compost piles and removed at different time intervals for germination rate determination. In a parallel experiment in the lab, P. aegyptiaca seeds were placed in incubators at fixed temperatures and removed periodically, to determine their germination rate. We found that ensuring compost temperatures above 55 °C for five to 6 h is sufficient to eradicate P. aegyptiaca seeds. This data was confirmed by the laboratory incubator experiments. Temperatures of 50–60 °C in the compost pile reduced C. campestris seed viability exponentially, but even 42 days of incubation did not completely eradicate their viability. Similar results were attained in a controlled temperature experiment in the laboratory, at which C. campestris seeds at 60 °C retained viability for 28 days. The results of this study stress the importance of maintaining high temperatures during manure composting procedures for eradication of P. aegyptiaca and C. campestris seeds and that. C. campestris seeds require a longer period of composting for eradication.