Progress in grafting watermelon to manage Verticillium wilt

Vegetable grafting for disease management was first used successfully when watermelon grafted onto a Cucurbita moschata rootstock overcame Fusarium wilt. Interspecific grafting has since been used effectively to mitigate several soilborne pathogens in a variety of solanaceous and cucurbitaceous cropping systems. Verticillium wilt caused by Verticillium dahliae is a significant disease in watermelon crops and is difficult to manage. Current management practices, including crop rotation, soil fumigation, and host resistance, are insufficient due to the ability of microsclerotia to persist in absence of a host, lack of efficacy of soil fumigants, and limited availability of resistant cultivars. Watermelon grafted onto commercial cucurbit rootstocks have increased tolerance to Verticillium wilt, although no cucurbit rootstocks are known to be completely resistant. Verticillium wilt incidence decreased on grafted plants grown in artificially and naturally infested soils, while scion health and growth as well as rootstock root mass and vigour increased. Commonly used rootstocks are Lagenaria siceraria, C. moschata, and C. maxima × C. moschata; of these, only C. maxima × C. moschata ‘Tetsukabuto’ reduced severity of Verticillium wilt across several scion cultivars, locations, years, and soil densities of V. dahliae. Although studies on Verticillium wilt resistance of grafted watermelon are few, their combined results suggest the threshold of V. dahliae soil density for watermelon may be around 5–12 cfu/g. This review summarizes available information on Verticillium wilt of watermelon and effects of different rootstock × scion combinations, assisting growers and breeding programmes in decisions to adopt watermelon grafting for management of Verticillium wilt.     

Characterization of resistance against the olive‐defoliating Verticillium dahliae pathotype in selected clones of wild olive

Verticillium wilt of olive is best managed by resistant cultivars, but those currently available show incomplete resistance to the defoliating (D) Verticillium dahliae pathotype. Moreover, these cultivars do not satisfy consumers' demand for high yields and oil quality. Highly resistant rootstocks would be of paramount importance for production of agronomically adapted and commercially desirable olive cultivars in D V. dahliae‐infested soils. In this work, resistance to D V. dahliae in wild olive clones Ac‐13, Ac‐18, OutVert and StopVert was assessed by quantifying the fungal DNA along the stem using a highly sensitive real‐time quantitative polymerase chain reaction (qPCR) protocol and a stem colonization index (SCI) based on isolation of V. dahliae following artificial inoculations under conditions highly conducive for verticillium wilt. Ac‐13, Ac‐18, OutVert and StopVert showed a symptomless reaction to D V. dahliae. The mean amount of D V. dahliaeDNA quantified in stems of the four clones ranged from 3.64 to 28.89 pg/100 ng olive DNA, which was 249 to 1537 times lower than that in susceptible Picual olive. The reduction in the quantitative stem colonization of wild olive clones by D V. dahliae was also indicated by a sharp decrease in the SCI. Overall, there was a pattern of decreasing SCI in acropetal progression along the plant axis, as well as correlation between positive reisolation and quantification of pathogen DNA. The results of this research show that wild olive clones Ac‐13, Ac‐18, OutVert and StopVert have a valuable potential as rootstocks for the management of verticillium wilt in olive.     

Present status of verticillium wilt of olive in Andalucía (southern Spain)1

Verticillium wilt caused by Verticillium dahliae is a disease highly prevalent in newly established olive orchards in Andalucía, southern Spain. Two syndromes of the disease occur in Andalucia, namely apoplexy and slow decline. Apoplexy is characterized by quick dieback of twigs and branches while slow decline consists of rapid drying out of inflorescences together with leaf chlorosis and necrosis. Systematic disease observations carried out in two experimental orchards planted with susceptible cv. Picual indicated that natural recovery of diseased trees occurred over time. Infection and vascular colonization of olive plants by V. dahliae were studied in susceptible (Picual) and resistant (Oblonga) cultivars inoculated with a mildly virulent or a highly virulent cotton-defoliating isolate of V. dahliae. Disease symptoms developed 24–32 days after inoculation in cv. Picual, but at that time plants of cv. Oblonga remained free from symptoms. However anatomical observations and isolations indicated that systemic infections by the two isolates had occurred to a large extent in both cultivars.     

Evaluation of resistance of Spanish olive cultivars to Verticillium dahliae in inoculations conducted in greenhouse

The resistance of 28 Spanish olive cultivars to Verticillium dahliae was evaluated in an experiment conducted under greenhouse conditions, by impregnating plant roots with a semisolid fluid mass of a mixture of culture medium and the conidia and mycelium of the fungus. Five-month-old olive plants were inoculated with a cotton defoliating isolate of V. dahliae. ‘Frantoio’ and ‘Picual’ were used as resistant and susceptible reference cultivars, respectively. Cultivars were assessed on the basis of final values of the area under the disease progress curve, mean severity of symptoms, and mortality at 26 weeks following inoculation. Verticillium wilt disease developed more slowly and reached lower values of these parameters than those normally recorded in previous studies conducted in growth chambers, using root-dip inoculation in a conidial suspension of the pathogen. However, most of the evaluated cultivars exhibited susceptible or moderately susceptible reactions to the infections caused by V. dahliae. In particular, a group of eight cultivars, from the same group as ‘Picual’, such as ‘Manzanilla de Abla’, ‘Manzanilla del Centro’ and ‘Negrillo de Iznalloz’, were significantly more susceptible than ‘Frantoio’. Conversely, ‘Escarabajillo’, ‘Menya’ and ‘Sevillana de Abla’ exhibited a high level of resistance to the disease, no dead plants, and vegetative recovery. Field experiments are currently being carried out to confirm the level of resistance assigned to these last genotypes. If confirmed, these genotypes will act as potential resistant genitors for inclusion in current olive breeding programs or for use as resistant rootstocks.     

Prospects and strategies in controlling verticillium wilt of olive1

Control of verticillium wilt of olive currently depends on preventive measures. Since systemic fungicides are unable to prevent or control the disease, its control should primarily be based on cultural methods, including irrigation systems which restrict dissemination of Verticillium dahliae propagules by irrigation water and avoidance of intercropping with hosts susceptible to V. dahliae. Since leaves from affected olive trees contribute, through formation of microsclerotia, to the inoculum in the soil, pruning should be practised prior to branch defoliation. As for resistant olive rootstocks or cultivars, promising verticillium-wilt resistance has been found in two rootstocks selected in California (US). However, these have to be tested under local conditions before they can be released to Mediterranean growers, while further search for other resistant rootstocks is needed. Soil solarization applied to individual diseased trees in established olive groves could substantially contribute to recovery or long-lasting symptom remission in the treated trees. This effect is mainly attributed to the decrease or eradication of V. dahliae microsclerotia in the treated soil but also to heat-tolerant fungal antagonists of the pathogen. Using herbicides to control weeds, and limiting soil rotovation, can restrict symptom development. Biological control can also be considered as a promising trend in controlling the disease by searching, testing and exploiting potential fungal or bacterial antagonists.     

Unravelling the relationships among Verticillium wilt,irrigation, and susceptible and tolerant olive cultivars

Survival, germination, olive colonization, and water-use efficiency (WUE) impairments by Verticillium dahliae could be influenced by cultivar susceptibility or irrigation, and this could modify the irrigation–pathogen–disease relationship. In this study, the combined effects of irrigation and cultivar susceptibility on Verticillium wilt (VW) development were modelled by the temporary assessment of V. dahliae propagules (total inoculum density, density of micropropagules, and sclerotia in wet and air-dried soil; ID, MpD, S_wD, and S_dD, respectively), root (RCI) and shoot (SCI) colonization indexes, and WUE. The relationship of disease severity to the measured parameters was then explored. Under controlled conditions, plants of cultivars ‘Picual’ and ‘Frantoio’ were irrigated to a high and low rate by varying drip-irrigation frequencies: daily, twice weekly, and a combination of daily for 11 days and then twice weekly. Disease severity and colonization parameters were higher in ‘Picual’, while WUE was higher in ‘Frantoio’. However, high rate and twice weekly and combination treatments significantly increased disease incidence and reduced time-to-symptoms-onset only in ‘Picual’, while high rate reduced WUE and increased relative ID, MpD, and S_wD in both cultivars. Irrigation did not affect SCI, but a higher RCI was found at high rate during the development of symptoms in ‘Picual’. By using classification trees to examine parameters—disease severity relationships, it was possible to determine the degree to which VW was affected by irrigation and/or cultivar susceptibility. MpD was the best indicator for VW detection at any time, WUE was best before symptoms developed, and RCI, total ID, and S_dD after symptoms developed.     

Interactions between Trichoderma harzianum and defoliating Verticillium dahliae in resistant and susceptible wild olive clones

Verticillium wilt (VW) in olive is best managed by an integrated disease management strategy, of which use of host resistance is a key element. The widespread occurrence of a highly virulent defoliating (D) Verticillium dahliae pathotype has jeopardized the use of commercial olive cultivars lacking sufficient resistance to this pathogen. However, the combined use of resistant wild olive rootstocks and Trichoderma spp. effective in the biocontrol of VW can improve the management of VW in olive. In vivo interactions between D V. dahliae and Trichoderma harzianum were studied in olive and wild olive plants displaying different degrees of resistance against this pathogen using confocal microscopy. This multitrophic system included wild olive clones Ac‐4 and Ac‐15, olive cv. Picual, and the fungal fluorescent transformants T. harzianum GFP22 and V. dahliae V138I‐YFP, the latter being obtained in this study. In planta observations indicated that V138I‐YFP colonizes the roots and stems of the olive and wild olive genotypes, and that GFP22 grows endophytically within the roots of them all. YFP fluorescence signal quantifications showed that: (i) the degree of root and stem colonization by the pathogen varied depending upon the susceptibility of the tested wild olive genotype, being higher in Ac‐15 than in Ac‐4 plants; and (ii) treatment with T. harzianum GFP22 reduced the extent of pathogen growth in both clones. Moreover, root colonization by strain GFP22 reduced the percentage of pathogen colonies recovered from stems of olive and wild olive plants.     

Evaluation of Olive Cultivars for Resistance to Verticillium dahliae

Resistance of 23 important olive cultivars to Verticillium dahliae has been evaluated in four experiments under controlled conditions. Nine-month-old nursery olive plants were inoculated with a cotton non-defoliating (ND) (V4) or a cotton defoliating (D) (V117) isolate of V. dahliae. Resistance was evaluated by assessing symptom severity using a 0–4 rating scale and estimating the area under disease progress curves. The percentage of plants killed and of those which recovered from the disease were used as additional parameters for including a particular cultivar into a defined category. Most of the evaluated cultivars were susceptible, although at different levels, to both isolates of V. dahliae. All cultivars were more susceptible to the D pathotype than to the ND one. A group of 11 cultivars, including several important Spanish cultivars, were susceptible or extremely susceptible to both pathotypes of V. dahliae. A second group showed differences of resistance depending on the pathotype used. They were susceptible or extremely susceptible to the D pathotype but resistant or moderately susceptible to the ND one. Finally, 'Frantoio', 'Oblonga' and 'Empeltre' were moderately susceptible to the D isolate of V. dahliae and resistant to the ND one. The resistance of 'Empeltre' was evident by the plant ability to recover from infection with either isolates. 'Empeltre' is considered to be a valuable cultivar for inclusion in breeding programmes for resistance to Verticillium wilt.     

Symptom development, pathogen isolation and Real-Time QPCR quantification as factors for evaluating the resistance of olive cultivars to Verticillium pathotypes

Verticillium wilt is the most serious olive disease in the Mediterranean countries and worldwide. The most effective control strategy is the use of resistant cultivars. However, limited information is available about the level and source of resistance in most of the olive cultivars and there are no published data using microsclerotia, the resting structures of Verticillium dahliae, as the infective inoculum. In the present study, we correlated symptomatology and the presence of the fungus along with the DNA relative amount (molecules μl⁻¹) of a defoliating (D) and a non-defoliating (ND) V. dahliae strain in the susceptible cv. Amfissis and the tolerant cvs Kalamon and Koroneiki, as quantified by the Real-Time QPCR technology. The viability of the pathogen in the plant tissues was confirmed by isolating the fungus on PDA plates, while symptom assessment proved the correlation between the DNA relative amount of V. dahliae in plant tissues and cultivar susceptibility. It was further demonstrated that the D and ND strains were present at a significantly higher level in cv. Amfissis than in cvs Kalamon and Koroneiki. It was finally observed that the relative amount of the pathogen in roots was lower than in stems and shoots and declined in plant tissues over time. These data constitute a valuable contribution in evaluating resistance of olive cultivars or olive root-stocks to V. dahliae pathotypes.     

Race 2 of Verticillium dahliae infecting tomato in Japan can be split into two races with differential pathogenicity on resistant rootstocks

Verticillium dahliae infecting tomato can be differentiated into races 1 and 2 based on differential pathogenicity on tomato cultivars carrying resistance gene Ve1. Although no commercial cultivars resistant to race 2 are available, race 2‐resistant rootstock cultivars Aibou and Ganbarune‐Karis have been bred in Japan. Nevertheless, the resistance of these rootstocks appears to be unstable in commercial tomato fields. Pathogenicity assays conducted under controlled conditions revealed that these rootstock cultivars are resistant to some isolates of race 2; this resistance is controlled by a single dominant locus, denoted by V2, based on segregation of resistance in F₂ populations from selfed rootstock cultivars. However, some other isolates of race 2 can overcome this resistance. Therefore it is proposed that the current race 2 of V. dahliae should be divided into two races, i.e. ‘race 2’ (nonpathogenic on Aibou) and ‘race 3’ (pathogenic on Aibou). The distribution of these races was surveyed in 70 commercial tomato fields in Hida, Gifu Prefecture, Japan. Race 3 was found in 45 fields, indicating that race 3 had already spread throughout the region. On the other hand, 25 fields had only race 2, and thus race 2‐resistant rootstocks would be effective for disease management in these fields. Races 2 and 3 could not be identified by genomic Southern hybridization probed with a telomere sequence, nor with previously reported race‐specific PCR assays. Elucidation of race‐determining mechanisms and development of methods for quick race identification should be made in future studies.     

Screening for resistance of Turkish olive cultivars and clonal rootstocks to Verticillium wilt

Verticillium wilt resistance of 77 olive cultivars including 71 domestic and six foreign ones, and four clonal rootstocks available in the olive gene bank, were tested using a highly virulent isolate (D pathotype) of Verticillium dahliae. The pathogen was stem-inoculated into the own-rooted saplings. Most cultivars and rootstocks were found to be extremely susceptible to the disease. ‘Sinop No. 1’, ‘E?riburun Nizip’, ‘Erkence’, ‘E?riburun Tatayn’, ‘Girit Zeytini’ and ‘Marantelli’ were highly resistant, as their disease severities did not exceed 10%. Additionally, 11 domestic cultivars (‘Sar? Habe?i’, ‘Ya?l?k Çelebi’, ‘Zoncuk’, ‘Dilmit’, ‘?am’, ‘Hurma Karaca’, ‘Erdek Ya?l?k’, ‘Melkabaz?’, ‘Yün Çelebi’, ‘Kan Çelebi’ and ‘Siyah Salamural?k’), two foreign cultivars (‘Arbequina’ and ‘Frantoio’) and one wild clonal rootstock (‘D36’) were found to be resistant, with disease severities less than 30%. On the other hand, the moderately susceptible group comprised ten domestic cultivars (‘Ak Zeytin’, ‘Ya? Çelebi’, ‘Saurani’, ‘Butko’, ‘Gemlik’, ‘Otur’, ‘Ya? Zeytini’, ‘Belluti’, ‘Sinop No. 2’ and ‘Samanl?’), three foreign cultivars (‘Leccino’, ‘Chemlali’ and ‘Ascolana’) and one wild clonal rootstock (‘D9’). The number of cultivars within highly resistant and resistant groups was 17 out of the 71 domestic cultivars from all regions (four from Aegean, seven from southeastern Anatolia, two from Black Sea and three from Marmara).     

Verticillium wilt of olive in Syria1

Verticillium wilt of olive was first recorded in Syria in mid-1978 and confirmed as due to Verticillium dahliae. This article reports observations made in a comprehensive survey of the disease in nine provinces over 7 years. Percent infection varied from 0.85 to 4.5 in different provinces. Newly planted groves in lowland areas showed more infection than older groves in hilly areas. Isolation of V. dahliae was possible at all times of year. Young trees were more susceptible to wilt than older ones. The performance of 13 local cultivars against wilt was studied under natural field conditions and found to vary greatly from susceptible to resistant. Agricultural practices greatly affect spread of the disease. High disease incidence was observed in irrigated groves compared with non-irrigated, and correlated positively with number of ploughings. Verticillium wilt causes a loss between 1 and 2.3% of total olive production annually.     

Novel methodologies in screening and selecting olive varieties and root-stocks for resistance to <Emphasis Type="Italic">Verticillium dahliae</Emphasis>

An innovative inoculation process, involving the drilling of a trunk hole in 3 year-old olive trees and injecting a dense conidial suspension of Verticillium dahliae, was developed to study differentiation in foliar symptom expression between olive cultivars tolerant or susceptible to the pathogen. It was demonstrated that V. dahliae conidia could be translocated and colonize the xylem at the same distance above and below the point of trunk injection in both cultivars. However, the pathogen could be subsequently isolated at statistically significant percentages in susceptible cv. Amphissis compared to the tolerant cv. Kalamon, indicating operation of resistance mechanisms in the vascular phase of the disease. Consequently symptom development in the susceptible cultivar was at least sixfold more intensive compared to the tolerant cultivar, 6–11 months after trunk inoculation. Perennial olive orchard experiments, aimed at selecting Verticillium-resistant root-stocks, were conducted by applying the novel method in 2–3 year-old root-stock suckers of Amphissis olive trees and in the tolerant cvs Lianolia of Corfu and Koroneiki. It was indicated that potentially resistant root-stocks could be obtained following the trunk drilling technique. Resistance differentiation between cvs Amphissis and Kalamon was further verified through root inoculation by various V. dahliae microsclerotial concentrations and demonstrated that the trunk drilling inoculation procedure is equally efficient in resistance evaluation of olives to Verticillium wilt. The trunk inoculation procedure could be useful in selecting and screening root-stocks for resistance to V. dahliae and other vascular pathogens and could elucidate resistance mechanisms in woody plants against vascular wilt diseases.     

Colonization in cotton plants by a green fluorescent protein labelled strain of Verticillium dahliae

Verticillium wilt of cotton (Gossypium hirsutum) is a widespread and destructive disease caused by the soil-borne fungal pathogen Verticillium dahliae. In this study, a green fluorescent protein (GFP) labelled V. dahliae strain (TV7) was obtained by transforming gfp into defoliating strain V991. Strain TV7 was used to study infection and colonization of wilt resistant cotton cultivar Zhongzhimian KV1 and susceptible cultivar 861 with the aid of confocal laser scanning microscopy. The results showed that initial infection and colonization of V. dahliae in Zhongzhimian KV1 and 861 were similar. Conidia and hyphal colonies formed and penetrated in the root meristematic and elongation zones and in the conjunction of the lateral and main roots. The invaded conidia started to germinate by 2 hpi (hours post-inoculation), penetrated into the root cortex and vascular bundles, eventually colonized in the stem xylem vessels and grew restrictedly in the individual tracheae of both resistant and susceptible cultivars. Moreover, pathogen DNA could be detected by qPCR in roots and stems of both cultivars, but its content in the wilt susceptible cultivar 861 was much higher than that in the wilt resistant cultivar Zhongzhimian KV1. The results indicated that the resistant cultivar has ability to suppress V. dahliae reproduction.     

Agricultural factors affecting Verticillium wilt in olive orchards in Spain

In recent years, the spread of Verticillium wilt in olive orchards, caused by the soil-borne pathogen Verticillium dahliae, is often related to intensive modern farming of highly productive cultivars, planted at high densities, usually irrigated, and under a mechanised system. The effects of agricultural factors associated with olive orchards were investigated in an important olive-growing area in southern Spain, as tools in predicting outbreaks of the disease. A stratified double-sampling technique was designed to determine the number of olive orchards needed to survey. A sampling survey was conducted from 2002 to 2005 in 873 olive orchards randomly selected, the owners were interviewed for details of agronomic factors, and orchards were inspected for the presence or absence of the disease. Polymerase chain reaction assays were carried out for identifying V. dahliae pathotypes. Pathogen prevalence showed a significant linear correlation with the mean plant density (r ² = 0.93), associated predominantly with a less virulent non-defoliating pathotype (r ² = 0.96). Overall, irrigation × high density caused disease incidence to peak in super-high-density olive-tree-planting systems. Olive orchards that had V. dahliae, however, did not differ in pathogen prevalence regardless of the olive cultivars. Young olive orchards were significantly more affected by V. dahliae than were old ones, particularly orchards with trees 8 to 12 years old. Irrigation increased pathogen prevalence and disease incidence in very young orchards (<7 years old). The prevalence of the non-defoliating pathotype was statistically high in young orchards whereas the prevalence of a highly virulent defoliating pathotype was high in old orchards.     

Sanitation of olive plants infected by Verticillium dahliae using heat treatments

Verticillium wilt, caused by the fungus Verticillium dahliae, is currently the most important disease affecting olive in the Mediterranean basin. There are no effective treatments for controlling this disease. The use of infected nursery stocks has largely contributed to the spread of the pathogen, and therefore the development of treatments to preventively sanitize the propagation stock is critical in the nursery industry. This study describes novel techniques to achieve this aim. The effects of several temperature–exposure time combinations were evaluated: (i) the survival of pathogen on culture medium (PDA); (ii) the pathogen viability on infected shoots and plants; (iii) the vegetative growth of plants of several cultivars; and (iv) the rooting ability of cuttings. The colonies of the pathogen growing in PDA were killed after 8 h and 60 min of exposure at 40 and 47°C, respectively. Temperatures ≥42°C for at least 2 h were lethal for the pathogen infecting the shoots. Likewise, moist hot air treatments at 42–44°C for 6–12 h eradicated the pathogen, without compromising the viability of the plants. Five olive cultivars were also evaluated and classified according to their thermotolerance as follows: sensitive (Chiquitita), moderately sensitive (Koroneiki, Frantoio and Picual) and heat tolerant (Arbequina). However, the optimized sanitation methods were applicable to all of the cultivars. Finally, heat treatments were applied to unrooted cuttings, which severely affected their rooting ability. Thus, this study developed a hot air treatment to produce V. dahliae‐free olive nursery plants.     

Assessment of the effect of surface drip irrigation on Verticillium dahliae propagules differing in persistence in soil and on verticillium wilt of olive

For efficient integrated management of verticillium wilt in olive (VWO), it is important to establish whether irrigation treatments (with Verticillium dahliae‐free water) that mitigate the disease in V. dahliae‐infested soil, also reduce the levels of more and less persistent propagules of the pathogen in the soil. Effects of irrigation on VWO and V. dahliae propagules were evaluated under natural environmental conditions. Potted plants were irrigated (pathogen‐free water) to two ranges of soil water content (RWC; high and low) at three surface drip‐irrigation frequencies (daily, weekly, and daily during some periods and otherwise weekly). VWO and total inoculum density (ID), density of less persistent micropropagules (MpD) and more persistent sclerotia in wet soil (S_wD), and sclerotia density for air‐dried soil (S_dD) were monitored. A logistic model (multiple sigmoid) of disease incidence revealed the lowest parameter values in treatments irrigated daily. Daily frequency of irrigation showed significantly lower disease incidence (39.2%) and disease intensity index (43.9%) and MpD (88.0%) values as areas compared with other frequencies, regardless of the RWC. High RWC significantly reduced (70.8–84.9%) ID, S_wD and S_dD as areas, but significantly increased (18.0%) the incidence of infected plants (IIP), regardless of the irrigation frequency. The disease incidence was not correlated with temperature. Daily irrigation to low RWC mitigated the VWO and the IIP, kept soil to the lowest MpD and resulted in the lowest S_dD level at the end of the trial. Results suggested that less persistent propagules could have played a part in the disease development.     

Development of a robust,VdNEP gene-based molecular marker to differentiate between pathotypes of Verticillium dahliae

Verticillium dahliae is a soilborne fungus that causes Verticillium wilt disease in a plethora of crops. Based on symptoms that develop on cotton, olive and okra, V. dahliae isolates are categorized into two pathotypes, namely defoliating and nondefoliating, with the former showing increased virulence and causing severe defoliation. Reliable differentiation between V. dahliae pathotypes is crucial for the management of Verticillium wilt in cotton and olive. In the present study, a polymorphism was detected among isolates of defoliating and nondefoliating pathotypes in Southern blots using the VdNEP gene as a probe. The regions flanking this gene were isolated by inverse PCR and sequence differences in the 3′ untranslated region (3′-UTR) of the VdNEP gene were detected between the two pathotypes. Based on these sequences, primers were designed and assessed to develop a multiplex PCR detection assay. Using this assay, a collection of cotton and olive V. dahliae isolates from Greece and Cyprus was screened, revealing that the defoliating pathotype is present in several regional units of Greece. Thus, this work presents a new, sensitive molecular marker for the differentiation between V. dahliae pathotypes based on the VdNEP gene. Because the 3′-UTR is involved in the phenotypes displayed by the pathotypes, an expression experiment was conducted under conditions simulating the xylem of a host plant. Expression of the VdNEP gene was elevated at all time points in the defoliating compared to the nondefoliating strain, suggesting a possible involvement of VdNEP expression in the defoliation process.     

Verticillium wilt of olive in Turkey: a survey on disease importance,pathogen diversity and susceptibility of relevant olive cultivars

A comprehensive survey on the prevalence and incidence of Verticillium wilt of olive in Turkey has been conducted over 6 years (2003–2008). Vegetative compatibility group (VCG) assessment and PCR-based molecular pathotyping were used to evaluate the distribution of the defoliating (D) and nondefoliating (ND) pathotypes of Verticillium dahliae in surveyed areas. Pathogen prevalence was 35% of all olive orchards inspected and incidence of the disease reached 3.1%. VCG1A was predominant (29.3%) and infected all major cultivars grown in Turkey. The other two VCGs detected (2A and 4B) were of minor relevance (4.9% and 0.9%, respectively). Disease incidence caused by VCG1A infections was higher (ranging from 1.1% to 6.9%) than that caused by VCG2A and VCG4B in 10 provinces (Manisa, Aydin, Kahramanmaras, Izmir, Mugla, Kilis, Denizli, Gaziantep, Mardin and Balikesir). However, VCG2A and 4B were more prevalent (and responsible for higher disease incidence) than VCG1A in three provinces (Hatay, Osmaniye and Bursa). Finally, VCG1A isolates were found in all provinces except Canakkale, and simultaneous presence of the three VCGs was only verified in Hatay province. An artificial inoculation bioassay (19 representative V. dahliae isolates included) revealed that VCG1A (13) isolates as a group were more aggressive and caused defoliation, whereas VCG2A (5) and VCG4B (1) isolates induced milder symptoms. Within a VCG group, virulence varied among isolates infecting the same olive cultivar and this virulence was also related to the differential susceptibility of the cultivars (‘Manzanilla’, ‘Ayvalik’ and ‘Gemlik’) tested. Molecular pathotyping allowed the identification of D (VCG1A) and ND (VCG2A/4B) pathotypes, which correlated with results from pathogenicity tests. Remarkably, the V. dahliae VCG1A/D pathotype population infecting olive in Turkey was molecularly different from that one previously identified in Spain.     

Cross-pathogenicity of <Emphasis Type="Italic">Verticillium dahliae</Emphasis> between potato and sunflower

This study examined cross-pathogenicity of the soilborne pathogen Verticillium dahliae between potato and sunflower. Four week-old potato and sunflower seedlings were inoculated with ten isolates from each of the two host species. Potato cultivars (Kennebec, susceptible, and Ranger Russet, moderately resistant) and sunflower hybrids (IS8048, susceptible, and 6946, moderately resistant) were assessed for disease severity and percent infection at 2 weeks, 3 weeks, 4 weeks, 5 weeks, and 6 weeks after inoculation (w.a.i), and for vascular discolouration at 6 w.a.i., using visual scales developed for each host species. The experiments were conducted in 2006 and repeated in 2007. Based on percent infection and disease severity, most V. dahliae isolates were highly aggressive on both host species. The tested isolates caused higher disease levels in the susceptible than in the moderately resistant phenotypes. They also caused more vascular discolouration in their original than in the alternative host. However, the isolates originating from sunflower caused less infection and disease severity on both hosts, compared to their potato counterparts. Cluster analysis based on all of the criteria used to assess pathogenicity led to three groups of isolates: (i) most V. dahliae potato isolates, which ranged with the highly aggressive control isolates, (ii) one V. dahliae sunflower isolate, which showed a similar pathogenicity level to the weakly-aggressive V. albo-atrum sub-group II control isolate, with no more symptoms than in the non-inoculated plants, and (iii) most V. dahliae sunflower isolates with mildly- to weakly-aggressive levels. Based on these results, V. dahliae cross-pathogenicity is very effective between potato and sunflower. Therefore, rotations involving these species should be avoided, especially where sunflower follows potato.