FUSARIUM Oxysporum f. sp. Ciceris (FOC) is the causal agent of FUSARIUM WILT, a destructive and widespread disease of chickpea. Rapid and accurate identification and detection of plant pathogens are essential for timely Disease Management (DM) strategies with appropriate measures. This study aimed to quantitatively determine FOC by using Quantitative Real-Time Polymerase Chain Reaction (qPCR) technique with specific primer pairs [Histone (H3) and Ribosomal (J5)] in seed, root, and root collar, and to discriminate it from other pathogenic fungi [FUSARIUM Oxysporum formae speciales (FO f. sp.) and Ascocyhta rabiei]. Total RNAs were isolated, converted to cDNAs (limit of 5 ng/rxn.-0.05 pg/rxn.) and used as template for qPCR studies. The FOC was detected in plant samples starting from the first day after inoculation. The FOC was detected in root, root collar and seed samples and was differentiated by qPCR assay from other pathogenic fungi. Melting curves, in which no primer dimers and non-specific complementation were observed, presented a single peak. Quantification was successfully performed using specific H3 and J5 primer pairs (P< 0.05), and the FOC was distinguished from other pathogenic fungi with J5 primer (P< 0.05). The results of these studies may support the development of new biochemical and molecular methods that allow direct, faster and more accurate determination of pathogens. Thus, it will also enable us to reduce the losses caused by diseases and the costs of DM.

Goudarzi, A. (2023). FUSARIUM WILT disease of date palm. Plant Pathology Science, 12(1), 64-73.        Abstract FUSARIUM WILT disease is known as one of the most destructive and fatal diseases of date trees in many date-producing areas of the world, especially date-producing countries in Africa. Currently, a significant portion of date orchards in Morocco and the western and central Sahara of Algeria are affected by FUSARIUM WILT disease. The economic losses caused by FUSARIUM WILT disease of dates, especially in the incidence of severe epidemics, are significant. Over the past century, the disease has killed more than 12 million palm trees in Morocco and about three million palm trees in Algeria. FUSARIUM oxysporum (Schltdl.) f. sp. albedinis, a soil-borne vascular pathogen, is known to cause FUSARIUM WILT disease of dates. The economic losses caused by this disease are very significant, especially in the incidence of severe epidemics, and in some areas, the FUSARIUM WILT epidemic has destroyed more than 70% of the date fruits. Integrated disease management strategies include the use of resistant or tolerant cultivars, prevention of introduction of the disease causal agent into disease-free areas, eradication of the disease, biological control and chemical control. This disease has not been reported from date-producing areas of Iran, although similar symptoms to FUSARIUM WILT have been observed in some date orchards of Hormozgan province.

FUSARIUM WILT of lettuce caused by the fungus FUSARIUM oxysporum f.sp. lactucum is a destructive disease of lettuce in fields of Tehran province. A glasshouse experiment was conducted to evaluate resistance and susceptibility of 20 lettuce cultivars obtained from, Denmark, Italy, England, USA and Iran to an aggressive isolate of the pathogen. The Index of percentage of seed germination, number of healthy plants, fresh and dry weight of shoots and dry weight of roots were used for cultivar evaluation. Results indicated that Salinas and Grand Rapids were the most resisitant cultivars base on the mentioned index (P<0.05). In the field experiment nine cultivars were planted in soil with 30% infection during the previous year. A Randomized Complete Block Design with four replications was used. Plants were inoculated around the roots at the two leaf stage with spore suspension of the pathogen isolate. Results indicated that local cultivars curl Ahwazi and Esfahani with no infection were resistant. Black Shahre-Ray and King Crown with 18.42% and 26.2% of infection respectively, showed an intermediate reaction to the pathogen.

Biocontrol of FUSARIUM oxysporum f. sp. ciceris by six Bacillus species was evaluated. Also plant growth promoting potential of the bacteria was assessed. Results showed that four bacterial strains produced the chitinase and cellulase enzymes and all isolates produced indole acetic acid. Bacillus licheniformis proved to be the most productive of hydrogen cyanide and particularly, Bacillus firmus solubilized phosphorus on Pikovskaya solid and liquid media. The majority of strains were able to produce siderophore and three produced NH3. Results showed that the Flip05-156C chickpea variety was less susceptible to Foc isolates compared to Flip93-93C and there was a clear difference in pathogenicity of the Foc isolates. Thus, Foc1 and Foc2 isolates caused 31. 25% and 41. 66%, plant mortality, respectively. As regards PGPR effect, results showed that B. licheniformis gave the best branching number, stem length and root weight of both chickpea varieties. However, Bacillus lentus distinctly improved the root length while Bacillus amyloliquefaciens improved weight of the vegetative parts.

Cotton (Gossypium hirsutumL. ) is one of the most significant cash crops as it is the backbone of the global textile industry. Although conventional breeding is already being used to solve the problems related to cotton, there is a rampant reduction in the yield and quality of cotton due to insect pests and fungal pathogens. The diseases that pose the major threat to cotton production are FUSARIUM and Verticillium WILTs. NaD1, a plant defense protein from Nicotian aalata, was found to have effective antifungal activity against many filamentous fungi including two of the major cotton pathogens, FUSARIUM oxysporum f. sp. Vasinfectum (Fov) and Verticillium dahlia. In the present study synthetic Nicotiana alata Defense 1 (NaD1) gene, under the control of a CaMV 35S constitutive promoter, was transferred into cotton, the transformation of shoot tip explants was achieved via Agrobacterium tumefaciens LBA4404. PCR and RT-PCR analyses showed that NaD1 was successfully incorporated into the cotton genome and expressed in transgenic lines. Comparing to the wildtype, NaD1-transgene cotton plants showed enhanced protection against V. dahlia. The success of this work may act as a stepping stone in the development of new cotton cultivars with improved fungal disease resistance and thus can play a major role in reducing loss due to the widely prevalent fungal diseases in cotton.

Introduction: WILT and root rot diseases caused by FUSARIUM oxysporum and FUSARIUM solani are among the most important diseases of chickpea throughout the world and Iran. However, there is not effective chemical fungicide against these soil-borne pathogenic fungi. In recent years, rhizospheric bacteria and fungi developed as promising biofungicides against soil-borne plant pathogens. These microbes exploit several mechanisms such as antibiotics, volatile organic compounds, sidereophore and induced systemic resistance to suppress pathogenic fungi. Recently, the cost of agrochemical innovation and the period of time for their registration have increased rapidly due to stringent legislation, both of these reasons favor investment in the manufacture of biopesticides. The annual progress rate of the biopesticide bazaar is more than 16%, but that of synthetic pesticides has been decreasing by 1. 5% annually. Biopesticide companies such as Agroquest, Gustafson, Marrone Bio Innovation, Certis, BioWorks, Becker Underwood, ABiTEP GmbH, and Prophyta have released effective biopesticides to the market. Agrochemical great-companies such as Bayer CropScience have also been engrossed in the bio-pesticide market. Bayer CropScience bought Agroquest in 2012 at a price of 425 million US dollars. Overall, biological control of plant pathogens is promising technology in management of plant disease in sustainable agriculture. Materials & Methods: In this study, potential of plant growth promoting rhizobacteria (PGPRs) were investigated on biological control of this diseases in laboratory and greenhouse condition. Soil samples were collected from chickpea fields in Kermanshah and Kurdestan provinces. The effect of isolated bacteria were assessed on mycelial growth of F. oxysporum and F. solani in dual culture test. The chickpea seeds were inoculated by 109 CFU/ml of each bacterial isolates and sown in pots. The effect of bacterial isolates have been investigated on plant growth factors in greenhouse situation. The same experiment was conducted to assess the biocontrol ability of selected isolates against FUSARIUM WILT and root rot disease of chickpea. The greenhouse experiments were conducted based on completely randomized design with four replicates. Data were subjected to analysis of variance procedure in SAS ver. 9. 1 software and means compression analysis was done by Fisher protected LSD. Finally, the 16S rDNA of the four selected isolates, B2, B3, B6 and B13 were sequenced and identified based on Genebank data. Results & Discussion: Sixteen out of 100 isolated inhibited the growth of both fungi in vitro. Isolates B13 with 11 mm inhibition zone had the highest growth inhibition activity against F. oxysporum. In absence of pathogens, B2 increased aerial part dry weight from 1. 08 to 3. 69 g in greenhouse condition. B3 and B4 were the best isolates in improving root growth. They increase root dry weight to 1. 42 and 1. 36 g, respectively. B2 and B13 isolates had the greatest effect on plant health and reduced disease severity up to 90% in F. oxysporum inoculated plants. The lowest biocontrol activity against F. oxysporum was recorded for isolate B6. B2 increased aerial part fresh weight from 0. 56 to 2. 49 g, root fresh weight from 0. 24 to 1. 31 g. B6 was the best isolate for suppression of F. solani and reduced the disease index by 73%. This isolate increased aerial organs fresh weight as up to 3. 2 folds and root fresh weight up to 2. 9 folds. However, all of bacteria isolates were able to reduce FUSARIUM root rot disease, significantly. Isolates B2, B3, B6 and B13 were characterized as Bacillus sp., Achromobacter sp., Bacillus pumilus and Burkholderia sp. based on 16S rDNA sequencing, respectively. Bacillus spp. strains exploit several mechanisms such as antibiosis, volatile organic compound production and induced systemic resistance against plant pathogens. These bacteria are good candidate to be formulated in spore suspension form. Here, Achromobacter introduced as good candidate for biological control of FUSARIUM diseases of chickpea for first time. Overall, plant probiotic bacteria could be consider as promising approach in management of chickpea FUSARIUM diseases. Conclusion: Bacterial isolates have different ability in plant growth improvement and suppression of plant pathogens. Bacillus sp. was the best isolate for enhancing the shoot dry weight while Achromobacter sp. was the best to improving root dry weight. Indeed, bacteria have several mechanisms for promoting plant growth. Auxin induction by volatiles of Bacillus spp. increase root while decreasing the shoot accumulation of auxin. Overall, consortia of bacteria strains seems to be promising approach to suppression of both diseases in chickpea.

Rasouli, L., Sadravi, M., & Keshavarz, K. (2023). Screening eight tomato varieties for resistance to FUSARIUM WILT disease. Plant Pathology Science, 12(1), 46-52.      Introduction: FUSARIUM WILT caused by soil-borne fungus FUSARIUM oxysporum f. sp. lycopersici (Fol) is one of the most important diseases of tomatoes in the world. Identification and cultivation of resistant varieties is an environmentally friendly method of disease management that leads to the production of a chemical-free and clean yield. This research was conducted to evaluate the reactions of eight tomato varieties to the disease and their correlation with peroxidase enzyme. Materials and Methods: WILTed tomato plants were sampled in the greenhouses of southwestern Iran and after isolation and purification and studying the morphological characteristics, two Fol  isolates were identified. The reactions of eight varieties vs. Kingston, Dafnis, Super chef, Karnak, CHpooya, Earlypooya, Superpooya, and CH to these two isolates were investigated in a factorial experiment with a completely randomized statistical design with four replications for each treatment under greenhouse conditions. The disease severity index and the vegetative and reproductive indices of the plant were determined. The level of peroxidase enzyme in the leaves of plants of each treatment was measured by spectrophotometric method at 470 nm wavelength and its correlation coefficient was determined with the disease severity index. Results: Analysis of the variance of these experimental data revealed a significant difference between Fol isolates in pathogenicity and interaction between Fol isolates and varieties. Fol2 isolate was more aggressive than Fol1 and CH was moderately susceptible, and  Earlypooya, Karnak, Superpooya, Dafnis, CHpooya, were moderately resistant, while Kingston and Super chef  were completely resistant to this hyper-virulent isolate of  the pathogen and disease. The peroxidase level had negatively correlated with the disease severity index. Conclusion: Among these varieties, Kingstone and Super chef are resistant to the disease. Peroxidase levels can be used as a resistance marker to assess how different tomato varieties react to the disease.

The FUSARIUM WILT of tomato which is caused by FUSARIUM oxysporumf.sp. lycopersici, is one of the most destractive diseases of tomato. In this research the effects of some fungicides, Ipredion +Carbendazim (Rovral TS), Thiobendazol (Tecto), Benlate (Benomyl), Topsin+ Thiram (Homai) and Thiophenate methyl (Topsin-M) on radial growth of FUSARIUM WILT of tomato in vitro and its pathogenicity in pot culture were studied under laboratory and greenhouse conditions. These fungicides showed 78-100 percent reduce growth of the pathogen when used in 30 ppm concentration in vitro Potato, Dextrose and Agar (PDA) after 7 days. The above fungicides in the concentrations of 100 and 1000 ppm were used in infested soil and as root drenched for seedlings respectively, in a completely randomized design with three replications. Results indicated that Rovral T-S decreased FUSARIUM WILT of tomato by 81.6% and 64.85% in pots of infested soil and seedling root dip respectively. Application of fungicides as seedling root dip and in pot of infested soil increased height and fresh weight of tomato shoots compared with the infested control. Rovral T-S fungicide with Ec50= 0.79 was recognized more suitable than the other fungicides in controlling the disease.  

Five isolates of Trichoderma (T. virens DAR 74290, T. harzianum T39, T. harzianum M,T. viride and Trichoderma sp.96) were evaluated for biocontrol of FUSARIUM WILT of melon (FUSARIUM oxysprum f.sp melonis). Dual culture and cellophane overlays techniques were used in vitro assay. Colony area was recorded, compared with controls and percentage inhibition of growth was calculated. Greenhouse experiments were performed to test the efficacy of Trichderma isolates for biological control of FUSARIUM WILT of melon. Colonized wheat bran with Trichoderma and infested wheat grain with pathogen were incorporated to potting mix as a rate of 10 and 14g per Kg respectively. Percentage of plants surviving determined 30 days after planting. Percentage of growth inhibition varied significantly among the antagonist isolate (P<0.01) and ranged from 0 to 100%. Cell free metabolite of T.virens DAR 74290 completely inhibited growth of the pathogen in vitro and appeared to be fungicidal. Percent of plants survival in pots treated with antagonist or healthy control was 100 and those of Trichoderma + pathogen varied from 5.3 to 61.06%. Population (cfu/g) of pathogen was stable and population of Trichoderma slightly decreased during the greenhouse experiments.