Revisiting strategies for reducing the seedbank of Orobanche and Phelipanche spp.

Weedy Orobanche and Phelipanche species are important in Southern and Eastern Europe, the Middle East and North Africa, and have recently been reported in the USA, Australia and some countries in South America. Rather than being controlled, the Orobanche and Phelipanche problem is increasing, both in intensity and in acreage. Large areas of new territory are at risk of invasion, if care is not immediately taken to limit the introduction of parasitic seeds and to educate farmers and others to be alert for new infestations. There is an urgent need to re-evaluate novel integrated Orobanche and Phelipanche management programmes that will allow a better control of the parasite species and limit their distribution. The main obstacle in the long-term management of Orobanche and Phelipanche infested fields is the durable seedbank, which may remain viable for decades in the field. Large quantities of long-lived seeds give the parasite genetic adaptability to changes in host resistance and cultural practices. As long as the seedbank is not controlled, the need to control the parasite will persist whenever a susceptible host is grown in the infested field. We discuss strategies for seedbank reduction, paying particular attention to cultural practices, whereas chemical and biological control methods are covered by other reviews in this issue.     

Temporal and spatial progress of the diseases caused by the crinivirus tomato chlorosis virus and the begomovirus tomato severe rugose virus in tomatoes in Brazil

Efficient management of whitefly-borne diseases remains a challenge due to the lack of a comprehensive understanding of their epidemiology, particularly of the diseases tomato golden mosaic and tomato yellowing. Here, by monitoring 16 plots in four commercial fields, the temporal and spatial distribution of these two diseases were studied in tomato fields in Brazil. In the experimental plots these diseases were caused by tomato severe rugose virus (ToSRV) and tomato chlorosis virus (ToCV), respectively. The incidence of each virus was similar in the plots within a field but varied greatly among fields. Plants with symptoms for both diseases were randomly distributed in three of four spatial analyses. The curves representing the progress of both diseases were similar and contained small fluctuations, indicating that the spread of both viruses was similar under field conditions. In transmission experiments of ToSRV and ToCV by Bemisia tabaci MEAM1 (former biotype B), these viruses had a similar transmission rate in single or mixed infections. It was then shown that primary and secondary spread of ToCV were not efficiently controlled by insecticide applications. Finally, in a typical monomolecular model of disease progress, simulation of the primary dissemination of ToSRV and ToCV showed that infected plants were predominantly randomly distributed. It is concluded that, although the manner of vector transmission differs between ToSRV (persistent) and ToCV (semipersistent), the main dispersal mechanisms are most probably similar for these two diseases: primary spread is the predominant mechanism, and epidemics of these diseases have been caused by several influxes of viruliferous whiteflies.     

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.     

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.     

Resistance of tomato accessions from the horticulture germplasm bank to red spider mites: exploring mite preference patterns and antixenosis mechanism

The red spider mite Tetranychus evansi can cause up to 90% yield losses to tomato crops. Mechanisms and causes of plant resistance to this pest could contribute to improved planning of plant breeding programs that prevent damage by this pest. It is known that the morphology and chemical compounds present in tomato plants are important to Tetranychus spp. success. This study aims at evaluating the resistance of 84 arbitrarily chosen tomato accessions – obtained from the Horticulture Germplasm of the Universidade Federal de Viçosa (HGB-UFV), Minas Gerais State, Brazil – to T. evansi. We evaluated the number of spider mites per leaf disk, number of trichomes/mm2, and the concentration of hydrocarbons of leaf disk. A significant difference in the number of T. evansi adults/leaf disk was found among accessions. The resistance mechanism of the HGB-UFV accessions was determined to be antixenosis. Accessions 2004, 2098, 2121, and 2100 had higher trichome density and the lowest adult T. evansi per tomato leaf disk. For this reason, these accessions should be chosen as sources of resistance in plant breeding programs. The hydrocarbons C₁₁, C₁₃, and C₂₂ had a negative correlation with T. evansi adults per leaf disk.     

Hormetic UV-C seed treatments for the control of tomato diseases

Hormesis is a dose response phenomenon in which low, non-damaging doses of a stressor bring about a positive response in the organism undergoing treatment. Evidence is provided here that hormetic UV-C treatments of tomato seed can control disease caused by Botrytis cinerea, Fusarium oxysporum f. sp. lycopersici (FOL) and f. sp. radicis-lycopersici (FORL) on tomato (Solanum lycopersicum). Treating seeds with a 4 kJ m⁻² dose of UV-C significantly reduced both the disease incidence and progression of B. cinerea, with approximately 10% reductions in both on cv. Shirley. Disease severity assays for FOL and FORL on cv. Moneymaker showed dose-dependent responses: UV-C treatments of 4 and 6 kJ m⁻² significantly reduced the disease severity scores of FOL, whilst only the 6 kJ m⁻² showed significant reductions for FORL. To determine the effects of treatment on germination and seedling growth, UV-C doses of 4, 8 and 12 kJ m⁻² were performed on cv. Shirley. No negative impacts on germination or seedling growth were observed for any of the treatments. However, the 8 kJ m⁻² treatment showed significant biostimulation, with increases in seedling, root and hypocotyl dry weight of 11.4%, 23.1% and 12.0%, respectively, when compared to the control. Furthermore, significant increases in the root-mass fraction (10.6%) and root:shoot ratio (13.1%) along with a decrease in shoot-mass fraction (2.0%) indicates that the 8 kJ m⁻² treatment stimulated root growth to the greatest extent. There was no effect on hypocotyl and primary root length or the number of lateral roots, indicating no adverse effects to basic root architecture or seedling growth.     

Genetic differentiation and host preference reveal non‐exclusive host races in the generalist parasitic weed Phelipanche ramosa

We developed 20 microsatellite markers to genotype over 100 populations of the parasitic weed Phelipanche ramosa, which covers a wide host crop and geographic range. A representative core collection of 15 populations was also used in cross‐infestation assays to study host preference during germination, attachment and shoot formation. We observed low genetic differentiation within most of the populations, but high genetic differentiation between populations partitioned into 3 genetic groups with different host preferences and geographic distributions. Genetic group 1 is detected exclusively in western France and on various host crops, notably winter oilseed rape (WOSR) and not hemp. Cross‐infection assays confirmed its incompatibility with hemp and showed its preference for WOSR and tobacco in terms of germination and attachment success. The group 2 populations share a large geographic distribution in France and Europe, low germination success with WOSR and high germination success, attachment success and shoot formation with hemp, tobacco or tomato. The subclades 2a and 2b include most of the French populations in hemp crops in eastern France and in tobacco fields in several European countries respectively. The genetic analyses revealed the potential of the three groups to increase their geographic range in the future. Intermediate genetic groups showed higher intrapopulation diversity and represent potential stocks for new host race emergence. Those findings argue in favour of the existence of host races in P. ramosa and should be considered for appropriate management strategies, notably in breeding programmes for resistance against this parasitic weed.     

Pepino mosaic virus isolates and differential symptomatology in tomato

Based on a survey conducted in commercial tomato production in Belgium in 2006, four Pepino mosaic virus (PepMV) isolates that differed in symptom expression in the crop of origin were selected for greenhouse trials. The selected isolates were inoculated onto tomato plants grown in four separate plastic tunnels. PepMV symptom development was assessed regularly and extensive sampling followed by ELISA analyses, genotyping and sequencing was performed to study viral presence and variation in PepMV sequences throughout the trial period. Two isolates (EU-mild and CH2-mild) that were selected based on mild symptom expression in the crop of origin caused only mild symptoms in the trial, while two other isolates (CH2-aggressive and EU + CH2) that were selected for severe symptom display, caused considerably more severe symptoms. Sequence homology between CH2-mild and CH2-aggressive was as high as 99·4%. Results of this study show that differential symptom expression can, at least partially, be attributed to the PepMV isolate, which may be related to minor differences at the nucleotide level between isolates.     

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.     

Comparison of three diagrammatic keys for the quantification of late blight in tomato leaves

Three diagrammatic grading keys were designed for the assessment of the severity of late blight (caused by Phytophthora infestans ) in tomato leaves. Simplified and broad keys considered, respectively, six (3, 12, 22, 40, 60 and 77%) and eight (3, 6, 12, 22, 40, 60, 77 and 90%) levels of disease severity, whilst a modified key based on a previous proposal for potato late blight considered six levels (1, 5, 10, 16, 32 and 50%). The keys were validated by 24 evaluators who assessed digital images of tomato leaves exhibiting different areas with lesions. Evaluator errors were compared using a mixed model in which evaluators were considered as random effects and the keys and evaluations as fixed effects. The accuracy and precision of the evaluators were compared by simple linear regression between the estimated and actual values of disease severity. The repeatability of evaluators was assessed using Pearson's correlation coefficient. There was significant ( P  ≤   0·001) variability amongst the errors made by evaluators, although the precision of each of the three keys was high with a coefficient of determination (R²) of 0·96, 0·93 and 0·83 for the simplified, broad and modified key, respectively. Repeatability of estimations amongst the evaluators was adequate (correlation coefficients of 0·91, 0·91 and 0·90 for the three keys, respectively). The simplified and broad keys resulted in higher precision and accuracy for the estimation of severity than did the modified key. Since the simplified key considers a smaller number of disease severity levels, its use is recommended in the assessment of late blight in tomato leaves.     

Serological and molecular detection of Tomato chlorosis virus and Tomato infectious chlorosis virus in tomato

Tomato chlorosis virus (ToCV) and Tomato infectious chlorosis virus (TICV) are two criniviruses inducing similar yellowing symptoms in tomato. An approximately 4 kb central region of the genomic RNA2 of French ToCV and TICV isolates was sequenced. TICV, for which no other sequences were available, appeared as a distant species in the genus, being close only to LIYV ( Lettuce infectious yellows virus ) for some, but not all, proteins. ToCV has more than 98% nucleotide identity with isolates from the US and Spain, and sequencing the CP gene of several isolates collected in different regions in southern France during 2 years suggested a unique origin. Polyclonal antisera were produced using capsid proteins of both viruses expressed in Escherichia coli . DAS-ELISA assays were developed for routine diagnosis and conditions for preparing samples for an optimized detection were determined. No cross-reactions were observed. However, some false-negative results, corresponding to samples giving ELISA readings close to the detection limit were regularly detected, particularly for ToCV (approximately 5% of the samples). A triplex RT-PCR assay was thus developed, which allowed detection of both viruses in a one-step protocol. An internal PCR control was included, which in addition showed that it could be used as a control for the entire RT-PCR procedure. Finally, combining DAS-ELISA in a first round, and triplex RT-PCR for doubtful samples, appeared the best way to achieve a reliable diagnosis of these viruses.     

Preventive and post‐infection control of Botrytis cinerea in tomato plants by hexanoic acid

The antifungal activity of hexanoic acid on the phytopathogen Botrytis cinerea was studied. This chemical inhibited both spore germination and mycelial growth in vitro in a concentration‐ and pH‐dependent manner, and stopped spore germination at a very early stage, preventing germ‐tube development. The minimum fungicidal concentration (MFC) for in vitro spore germination was 16 mm . Hexanoic acid also inhibited in vitro mycelial growth of germinated spores at an MFC of 12 mm . Studies performed to characterize the mechanisms underlying the antimicrobial effect of hexanoic acid showed that it alters fungal membrane permeability. In addition, hexanoic acid treatment increased the levels of spermine, spermidine, putrescine and cadaverine in B. cinerea mycelia. Spray application of hexanoic acid at fungicidal concentrations on 4‐week‐old tomato plants prior to fungal inoculation reduced necrosis diameter by approximately 60%. Application of the same hexanoic acid concentrations on previously infected plants reduced further necrosis expansion by around 30%. The results suggest that this chemical acts as a preventive and curative fungicide. Interestingly, treatments with hexanoic acid at concentrations below the MFC in hydroponic solution prior to fungal inoculation significantly reduced necrosis area. These results suggest an inducer effect of plant responses for hexanoic acid treatments at these concentrations. Hexanoic acid is a good candidate for safe antifungal treatments for the control of B. cinerea, which is responsible for many economic losses on fruits, vegetables and flowers.     

中国特色蝗灾治理技术体系及应用成效

中国有数千年蝗灾发生为害的历史,建国初期在政府的组织领导下,在科研人员和广大群众的努力下,提出并实施了“改治并举”的策略,改造东亚飞蝗发生基地近2/3,同时大规模使用化学农药进行防治,初步形成了以改造蝗虫发生基地为特色的蝗灾治理技术体系,经过长期实践,蝗灾得到有效遏制,保障了我国粮食生产安全和社会稳定,取得了世界瞩目的成绩。自20世纪80年代起,随着利用蝗虫微孢子虫Nosema locustae和绿僵菌Metarhizium spp.等微生物的治蝗技术获得突破性进展,我国已经形成了以生态控制（改造蝗虫发生基地）为基础,以生物防治为核心,以信息技术为辅助的具有中国特色的蝗灾治理技术体系,并走在国际前列。该技术体系的长期大规模应用,不但在有效防治蝗灾保障我国粮食安全生产、避免大规模使用化学农药造成环境污染和生态失衡等方面发挥了重大作用,而且也在支援国际蝗灾治理中做出了贡献。最后对未来蝗灾治理的关键问题和发展方向进行了探讨。