Role of temperature, moisture and Trichoderma species on the survival of Fusarium oxysporum ciceri in the rainfed areas of Pakistan

It has been observed in the present study that when spores of Trichoderma harzianum (Th-2) isolate were applied in the sandy clay loam soil and continuously incubated for 4 months at 25 degrees C and 35 degrees C and at three water potentials, -0.03 MPa, -0.3 MPa and <-50 MPa, it has resulted in significantly reduced (P<0.05), growth of Fusarium oxysporum ciceri (Foc) on branches of chickpea plant. The pathogen population was greatly reduced in the moist soil (43 MPa) when compared with the wet soil (-0.03 MPa) at both temperatures which was indicated by greater colonization and growth of T. harzanum-2 on the branch pieces of chickpea plants. The pathogen was completely eradicated from the chickpea branch pieces, after 6 months at 35 degrees C in the moist soil. In air-dry soil (<-50 MPa), Foc survived in 100% of the branch pieces even after 6 months at both temperatures. When chickpea plant branch pieces having pathogen was sprayed with Th-2 antagonistic isolates of Trichoderma spp., the Th-2 isolate killed the pathogen up to minimum level (10-12%) after 5 months at 35 degrees C in the sandy clay loam soil. It can be concluded that in chickpea growing rainfed areas of Pakistan having sandy clay loam soil, Foc can be controlled by using specific Trichoderma spp., especially in the summer season as after harvest of the crop the temperature increased up and there is rainfall during this period which makes the soil moist. This practice will be able to reduce the inoculum of Foc during this hot period as field remain fallow till next crop is sown in most of the chickpea growing rainfed areas of Pakistan.

Comparative effect of root-knot nematode on severity of Verticillium and Fusarium wilt in cotton

The effect of root-knot nematode (RKN) (Meloidogyne incognita) on Verticillium dahliae and Fusarium oxysporum f.sp. vasinfectum in cotton (Gossypium hirsutum) was investigated. Two different inoculation methods were used, one in which inoculum was added to the soil, so that nematode and fungal inoculum were in close proximity; the other, inoculation into the stem, whereby the two inocula were spatially separated. Invasion of the roots by RKN enhanced disease severity, as measured by the height of vascular browning in the stem, following inoculation with either wilt pathogen. The effect of RKN on Fusarium wilt was more pronounced than that on Verticillium wilt. Nematode-enhanced infection by F. oxysporum is a well known effect but there are few reports of enhanced infection by Verticillium due to RKN. Relative resistance of a number of cotton cultivars to both wilt diseases, as measured by height of vascular browning, was similar to the known field performance of the cultivars. The use of vascular browning as an estimate of disease severity was therefore validated.

Enhancement of germination of spores of Verticillium dahliae and Fusarium oxysporum esp vasinfectum in vascular fluid from cotton plants infected with the root-knot nematode

Root-knot nematode [RKN] (Meloidogyne incognita) can increase the severity of Verticillium (V dahliae) and Fusarium (F oxysporum f.sp. vasinfectum) wilt diseases in cotton (Gossypium hirsutum). This study was conducted to determine some of the physiological responses caused by nematode invasion that might decrease resistance to vascular wilt diseases. The effect of RKN was investigated on spore germination and protein, carbohydrate and peroxidase content in the xylem fluids extracted from nematode-infected plants. Two cotton cultivars were used with different levels of resistance to both of the wilt pathogens. Spore germination was greater in the xylem fluids from nematode-infected plants than from nematode-free plants. The effect on spore germination was greater in the Fusarium-resistant cultivar (51%). Analysis of these fluids showed a decrease in total protein and carbohydrate levels for both wilt-resistant cultivars, and an increase in peroxidase concentration. Fluids from nematode-free plants of the Verticillium-resistant cultivar contained 46% more peroxidase than the Fusarium-resistant cultivar. The results provide further evidence that the effect of RKN on vascular wilt resistance is systemic and not only local. Changes in metabolites in the xylem pass from the root to the stem, accelerating disease development.

Improving the affinity of tomato grafted on Solanum torvum using an intermediate rootstock

Most of the interspecific rootstocks (Lycopersicum esculentum × L. hirsutum) used in grafted Spanish tomato crops are resistant to Meloidogyne nematodes, but the 'Mi' resistance gene does not work well at high soil temperatures. Ralstonia solanacearum is a bacterial disease usual in tropical areas, but recently identified with low incidence in several European countries. This disease could be controlled by grafting tomato on Solanum torvum, which is also resistant to Meloidogyne. However, S. torvum and tomato have low grafting affinity, which could be improved using an intermediate rootstock. Some cultivars of eggplant have a relatively good affinity with tomato and complete affinity with S. torvum. In this study we compared two tomato cultivars (one resistant to Verticillium dalihae, Fusarium oxysporum v. lycopersici race 2 and Meloidogyne spp., and one non-resistant) grafted onto 'Beaufort' (Lycopersicum esculentum × L. hirsutum), 'Torvum Vigor' (Solanum torvum) and also with an intermediate grafting of eggplant ('Cristal') between tomato and S. torvum, with nongrafted plants as controls. This arrangement was carried out in two cropping cycles (winter-spring and summer-autumn). In both cycles, plants grafted onto S. torvum, both single or double grafted, yielded less than those grafted onto 'Beaufort' or nongrafted plants. In the spring cycle, no differences were found between single and double-grafted plants using S. torvum rootstocks, but in the autumn cycle double grafted plants had higher yields than the single grafted plants. The severity of nematode infections, in terms of reducing yields, and/or hypothetical infections of Ralstonia, will determine the utility of this technique in tomato production.

Fusarium solani f. sp. passiflorae: A new forma specialis causing collar rot in yellow passion fruit

The aim of this study was to characterize a Fusarium population obtained from yellow passion fruit (YPF) with collar rot using pathogenicity, morphocultural characteristics and molecular tests. Pathogenicity and disease severity were assessed in six plant species: YPF, zucchini, tomato, bean, soya bean and cucumber. Potato dextrose agar medium (PDA) was used to determine mycelial growth at five temperatures (15-35°C). The colour produced by isolates was also determined on PDA at 25°C. Synthetic nutrient agar medium was used to evaluate: (i) type of mycelium and phialides; (ii) size, shape and number of septa from conidia; and (iii) production of chlamydospores and perithecia. Molecular tests consisted of sequencing the ITS-5·8S rDNA region and elongation factor 1α (EF-1α) gene. The isolates caused large lesions on YPF, zucchini and tomato, with YPF having the highest mean disease severity and being the only one that showed wilt symptoms and death of the plant. Thus the isolates showed host specificity. Maximum mycelial growth occurred at 25°C and the predominant colour was bluish-white. The isolates produced long phialides, dense aerial mycelium, oval microconidia with a mean size of 9·5 × 2·6 μm, macroconidia of 32·7 × 3·4 μm with 3·3 septa, and chlamydospores; only one isolate lacked perithecia. Phylogenetic trees of the ITS region and EF-1α gene showed that isolates from YPF formed a distinct group within the F. solani group and the formae speciales of F. solani. It is proposed to name all isolates from YPF as F. solani f. sp. passiflorae. © 2013 British Society for Plant Pathology.

Influência dos metabólitos secundários de Piper divaricatum da região amazônica no controle do Fusarium solani f. sp. piperis causador da fusariose em pimenta do reino

A cultura de pimenta do reino (Piper nigrum L.) é uma das principais atividades agrícolas no estado do Pará e sofre sérios danos ocasionados pela fusariose, doença restrita ao Brasil. O presente trabalho avaliou a atividade antifúngica in vitro de óleos essenciais de espécies de Piper ricos em fenilpropanóides frente a Fusarium solani f. sp. piperis, agente causal da fusariose. A inibição do crescimento micelial pelo método de difusão em Agar na concentração de 5 mg.Ml^-1 foi considerada: baixa para P. aduncum (20,3%) e P. krukoffii (31,4%); moderada para P. callosum (55,7%) e P. marginatum (70,3%) e alta para P. divaricatum (93,3%). Os componentes majoritários identificados por CG-EM foram: dilapiol (92,0%), safrol (78,0%), metileugenol (75,2%) e eugenol (7,9%), apiol (80,0%), Z-isoosmorizol (44,0%) e E-anetol (22,0%), respectivamente. O óleo de P. divaricatum e seus compostos majoritários apresentaram valores CIM de 0,75 mg.mL^-1. A avaliação dos efeitos combinados do eugenol e metileugenol apontou o eugenol como principal responsável pela atividade.

Influência de fungos micorrízicos arbusculares associa- dos ou não a Fusarium oxysporum Schecht. sobre plantas de alecrim (Rosmarinus officinalis L.) e manjericão (Ocimum basilicum L.)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Desenvolvimento de substrato supressivo à murcha do crisântemo causada por Fusarium oxysporum

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Bioactive extracts and chemical constituents of two endophytic strains of Fusarium oxysporum

Ethyl acetate extracts of cultures grown in liquid Czapek and on solid rice media of the fungal endophyte Fusarium oxysporum SS46 isolated from the medicinal plant Smallanthus sonchifolius (Poepp.) H. Rob., Asteraceae, exhibited considerable cytotoxic activity when tested in vitro against human cancer cells. Chromatographic separation yielded anhydrofusarubin (1) and beauvericin (2) that were identified based on their ¹H and 13C NMR data. Compounds 1 and 2 showed the strongest cytotoxic activity against different cancer cell lines. Compound 2 also showed promising activity against Leishmania braziliensis. Hexanic extract of F. oxysporum SS50 grown on solid rice media also afforded a mixture of compounds that displayed cytotoxic activity against different cancer cell lines. Chemical analysis of the mixture of compounds, investigated by gas chromatography-mass spectrometry (GC-MS), showed that there was a predominance of methyl esters of fatty acids and alkanes.

Mezzi innovativi nella difesa delle cucurbitacee da Fusarium solani f.sp. cucurbitae

Lo studio “Lotta biologica a Fusarium solani f.sp. cucurbitae su zucchino” si colloca nell’ambito della difesa integrata delle colture orticole dalle fitopatie fungine, in particolare quelle causate da patogeni ad habitat terricolo nei confronti dei quali è sempre più frequente il ricorso a mezzi di lotta diversi dai prodotti chimici. Interessante e innovativa appare la prospettiva di utilizzare microrganismi adatti a svilupparsi nel suolo, competenti per la rizosfera delle piante e con spiccate caratteristiche d’antagonismo verso i patogeni tellurici e di stimolazione delle difese sistemiche della pianta. Il marciume del colletto delle cucurbitacee, causato da diversi patogeni tra cui Fusarium solani f.sp. cucurbitae, rappresenta la principale malattia fungina di tipo tellurica che colpisce lo zucchino ed il melone nella Pianura Padana e che può portare a consistenti perdite produttive. Indagini condotte dal 2004 da parte del Diproval nell’areale bolognese, hanno evidenziato un’elevata frequenza del patogeno su zucchino coltivato soprattutto in tunnel. Considerata la carenza di conoscenze locali di F. solani f.sp. cucurbitae e di mezzi chimici di lotta efficaci, la ricerca svolta ha inteso approfondire la diagnosi della malattia e le caratteristiche biologiche degli isolati locali, e valutare la possibilità di utilizzare metodi biologici di lotta contro questo patogeno, nonché di studiare alcune delle possibili modalità d’azione di microrganismi antagonisti. Sono state pertanto prelevate, da zone diverse del Bolognese, campioni di piante di zucchino che presentavano sintomi di marciume del colletto ed è stato isolato il patogeno, che in base alle caratteristiche morfologiche macro e microscopiche, alle prove di patogenicità su diversi ospiti e a saggi biomolecolari, è stato identificato come Fusarium solani f. sp. cucurbitae W.C. Snyder & H.N. Hansen razza 1. Dagli isolati di campo sono state realizzate un centinaio di colture monosporiche venti delle quali sono state utilizzate per la prosecuzione delle prove. I venti ceppi sono stati saggiati per la loro patogenicità inoculandoli in terriccio sterile e con trapianto di giovani piante di zucchino. E’ risultata un’elevata variabilità del livello di virulenza tra i ceppi, stimata da 39% a 83% riguardo la gravità della malattia e da 61 a 96% per la frequenza di malattia. Sono state condotte prove di accrescimento miceliare che hanno evidenziato differenze tra i ceppi e tra gli esperimenti condotti a tre diverse temperature (17, 23 e 28°C) alla luce ed al buio. La crescita maggiore complessivamente è stata ottenuta a 23°C. I venti ceppi hanno anche mostrato di produrre, in vitro, vari livelli di enzimi di patogenesi quali cellulasi, poligalatturonasi, pectin liasi e proteasi. E’ stata evidenziata unan correlazione significativa tra attività cellulasica e pectin liasica con frequenza e gravità della malattia dei venti ceppi del patogeno. Le prove relative al contenimento della malattia sono state condotte in cella climatica. Sono stati considerati prodotti commerciali (Remedier, Rootshield, Cedomon, Mycostop, Proradix, Clonotry) a base rispettivamente dei seguenti microrganismi: Trichoderma harzianum ICC012 + T. viride ICC080, T. harzianum T22, Pseudomonas chlororaphis MA342, Streptomyces griseoviridis K61, P. fluorescens proradix DSM13134 e T. harzianum + Clonostachys rosea). I prodotti sono stati somministrati sul seme, al terreno e su seme+terreno (esperimenti 1 e 2) e in vivaio, al trapianto e vivaio+trapianto (esperimenti 3 e 4), riproducendo situazioni di pratico impiego. L’inoculazione del patogeno (un ceppo ad elevata patogenicità, Fs7 ed uno a bassa patogenicità, Fs37) è stata effettuata nel terreno distribuendo uno sfarinato secco di semi di miglio e cereali colonizzati dal patogeno. La malattia è stata valutata come intensità e gravità. I prodotti sono stati impiegati in situazioni di particolare stress, avendo favorito particolarmente la crescita del patogeno. Complessivamente i risultati hanno evidenziato effetti di contenimento maggiore della malattia nel caso del ceppo Fs37, meno virulento. La malattia è stata ridotta quasi sempre da tutti i formulati e quello che l’ha ridotta maggiormente è stato Cedomon. Il contenimento della malattia somministrando i prodotti solo nel terreno di semina o di trapianto è stato in generale quello più basso. Il contenimento più elevato è stato ottenuto con la combinazione di due tipologie di trattamento, seme+terreno e vivaio+trapianto. Le differenze tra i prodotti sono risultate più evidenti nel caso del ceppo Fs7. Per quanto riguarda lo studio di alcune delle modalità d’azione dei microrganismi contenuti nei formulati più efficaci, è stato verificato che tutti sono stati in grado di inibire, se pur in vario modo, la crescita del patogeno in vitro. Gli antagonisti più efficaci sono stati S. griseoviridis K61 (Mycostop) e P. fluorescens proradix DSM13134). I ceppi di Trichoderma, ed in particolare T.harzianum T22 (Rootshield), sono risultati i più attivi colonizzatori del substrato. Riguardo il fenomeno dell’antibiosi, il batterio P. fluorescens proradix DSM13134 ha mostrato di produrre i metaboliti non volatili più efficaci nel ridurre lo sviluppo del patogeno. Nelle condizioni sperimentali adottate anche i due ceppi di T. viride ICC080 e T. harzianum ICC012 hanno dimostrato di produrre metaboliti efficaci. Tali risultati, anche se relativi a prove in vitro, possono contribuire alla comprensione dei meccanismi dei microrganismi sul contenimento dell’infezione relativamente al rapporto diretto sul patogeno. E’ stato inoltre verificato che tutti i microrganismi saggiati sono dotati di competenza rizosferica e solo i batteri di endofitismo. Si conclude che, nonostante l’elevata pressione infettiva del patogeno che ha certamente influito negativamente sull’efficacia dei microrganismi studiati, i microrganismi antagonisti possono avere un ruolo nel ridurre l’infezione di F. solani f.sp. cucurbitae razza 1.

Salicylic acid mediates resistance to the vascular wilt pathogen Fusarium oxysporum in the model host Arabidopsis thaliana

Fusarium oxysporum is a soilborne fungal pathogen that causes major economic losses by inducing necrosis and wilting symptoms in many crop plants. In this study, the interaction between F. oxysporum and the model plant Arabidopsis thaliana has been investigated to better understand the nature of host defences that are effective against the Fusarium wilt pathogen. The expression of salicylate- and jasmonate-responsive defence genes in F. oxysporum-challenged roots of A. thaliana plants as well as in the roots of plants whose leaves were treated with salicylate or jasmonate was analysed. Unexpectedly, genes (e.g. PR1, PDF1.2, and CHIB) encoding proteins with defensive functions or transcription factors (e.g. ERF1, AtERF2, AtERF4 and AtMYC2) known to positively or negatively regulate defences against F. oxysporum were not activated in F. oxysporum-inoculated roots. In contrast, the jasmonate-responsive defence gene PDF1.2 was induced in the leaves of plants whose roots were challenged with F. oxysporum, but the salicylate- responsive PR1 gene was not induced in the leaves of inoculated plants. Exogenous salicylic acid treatment prior to inoculation, however, activated PR1 and BGL2 defence gene expression in leaves and provided increased F. oxysporum resistance as evidenced by reduced foliar necrosis and plant death. Exogenous salicylic acid treatment of the foliar tissue did not activate defence gene expression in the roots of plants. This suggests that salicylate- dependent defences may function in foliar tissue to reduce the development of pathogen-induced wilting and necrosis. Despite the induction of defence gene expression in the leaves by jasmonate, this treatment did not lead to increased resistance to F. oxysporum. Overall, the results presented here suggest that the genetic manipulation of plant defence signalling pathways is a useful strategy to provide increased Fusarium wilt resistance.

Efeito direto de fosfitos de potássio a Fusarium solani f. sp. cucurbitae.

O presente trabalho objetivou avaliar o efeito de doses (1,25; 2,5; 5,0 e 10 mL L-1) de fosfito de potássio no crescimento micelial e na germinação de conídios de Fusarium solani f. sp. cucurbitae, agente causal da podridão-do-colo e raízes do meloeiro. Foram avaliadas as doses 1,25; 2,5; 5,0 e 10 mL L-1 dos fosfitos, uma testemunha apenas com água (dose ?0?) e o fungicida tebuconazol + trifloxistrobina (1,0 mL L-1). As formulações de fosfito de potássio avaliadas proporcionaram toxidez direta a F. solani f. sp. cucurbitae, apresentando efeito significativo de doses no crescimento micelial e germinação de conídios do fungo. Contudo, observou-se maior efeito das formulações de fosfitos na inibição da germinação (DL50 =1,7 e 1,1 mL-1) do que na inibição do crescimento micelial (DL50 >10 mL-1 para os dois fosfitos). Na maior dose avaliada (10 mL L-1), a inibição do crescimento micelial foi de 33% e 18%, na dose de 5 mL L-1 as formulações inibiram a germinação em 88% e 89%. O fungicida inibiu totalmente o crescimento micelial e em inibiu a germinação dos conídios em torno de 99%, diferindo dos demais tratamentos.

Global banana disease management - getting serious with sustainability and food security

Much research in understanding plant diseases has been undertaken, but there has been insufficient attention given to dealing with coordinated approaches to preventing and managing diseases. A global management approach is essential to the long-term sustainability of banana production. This approach would involve coordinated surveys, capacity building in developing countries, development of disease outbreak contingency plans and coordinated quarantine awareness, including on-line training in impact risk assessment and web-based diagnostic software. Free movement of banana plants and products between some banana-producing countries is causing significant pressure on the ability to manage diseases in banana. The rapid spread of Fusarium oxysporum f. sp. cubense 'tropical race 4' in Asia, bacterial wilts in Africa and Asia and black leaf streak [Mycosphaerella fijiensis] in Brazil and elsewhere are cases in point. The impact of these diseases is devastating, severely cutting family incomes and jeopardising food security around the globe. Agreements urgently need to be reached between governments to halt the movement of banana plants and products between banana-producing countries before it is too late and global food security is irreparably harmed. Black leaf streak, arguably the most serious banana disease, has become extremely difficult to control in commercial plantations in various parts of the world. Sometimes in excess of 50 fungicide sprays have to be applied each year. Disease eradication and effective disease control is not possible because there is no control of disease inoculum in non-commercial plantings in these locations. Additionally, there have been enormous sums of money invested in international banana breeding programmes over many years only to see the value of hybrid products lost too soon. 'Goldfinger' (AAAB, syn. 'FHIA-01'), for example, has recently been observed severely affected by black leaf streak in Samoa. Resistant cultivars alone cannot be relied upon in the fight against this disease. Real progress in control may only come when the local communities are engaged and become actively involved in regional programmes. Global recommendations are long overdue and urgently needed to help ensure the long-term sustainable utilisation of the products of the breeding programmes.

Organic inputs, tillage and rotation practices influence soil health and suppressiveness to soilborne pests and pathogens of ginger.

A field experiment was established in which an amendment of poultry manure and sawdust (200 t/ha) was incorporated into some plots but not others and then a permanent pasture or a sequence of biomass-producing crops was grown with and without tillage, with all biomass being returned to the soil. After 4 years, soil C levels were highest in amended plots, particularly those that had been cropped using minimum tillage, and lowest in non-amended and fallowed plots, regardless of how they had been tilled. When ginger was planted, symphylans caused severe damage to all treatments, indicating that cropping, tillage and organic matter management practices commonly used to improve soil health are not necessarily effective for all crops or soils. During the rotational phase of the experiment, the development of suppressiveness to three key pathogens of ginger was monitored using bioassays. Results for root-knot nematode (Meloidogyne javanica) indicated that for the first 2 years, amended soil was more suppressive than non-amended soil from the same cropping and tillage treatment, whereas under pasture, the amendment only enhanced suppressiveness in the first year. Suppressiveness was generally associated with higher C levels and enhanced biological activity (as measured by the rate of fluorescein diacetate (FDA) hydrolysis and numbers of free-living nematodes). Reduced tillage also enhanced suppressiveness, as gall ratings and egg counts in the second and third years were usually significantly lower in cropped soils under minimum rather than conventional tillage. Additionally, soil that was not disturbed during the process of setting up bioassays was more suppressive than soil which had been gently mixed by hand. Results of bioassays with Fusarium oxysporum f. sp. zingiberi were too inconsistent to draw firm conclusions, but the severity of fusarium yellows was generally higher in fumigated fallow soil than in other treatments, with soil management practices having little impact on disease severity. With regard to Pythium myriotylum, biological factors capable of reducing rhizome rot were present, but were not effective enough to suppress the disease under environmental conditions that were ideal for disease development.

Ground cover management alters development of Fusarium wilt symptoms in Ducasse bananas

Many banana producing regions around the world experience climate variability as a result of seasonal rainfall and temperature conditions, which result in sub-optimal conditions for banana production. This can create periods of plant stress which impact on plant growth, development and yields. Furthermore, diseases such as Fusarium wilt caused by Fusarium oxysporum f. sp. cubense, can become more predominant following periods of environmental stress, particularly for many culturally significant cultivars such as Ducasse (synonym Pisang Awak) (Musa ABB). The aim of this experiment was to determine if expression of symptoms of Fusarium wilt of bananas in a susceptible cultivar could be explained by environmental conditions, and if soil management could reduce the impact of the disease and increase production. An experiment was established in an abandoned commercial field of Ducasse bananas with a high incidence of Fusarium wilt. Vegetated ground cover was maintained around the base of banana plants and compared with plants grown in bare soil for changes in growth, production and disease symptoms. Expression of Fusarium wilt was found to be a function of water stress potential and the heat unit requirement for bananas. The inclusion of vegetative ground cover around the base of the banana plants significantly reduced the severity and incidence of Fusarium wilt by 20 % and altered the periods of symptom development. The growth of bananas and development of the bunch followed the accumulated heat units, with a greater number of bunched plants evident during warmer periods of the year. The weight of bunches harvested in a second crop cycle was increased when banana plants were grown in areas with vegetative ground cover, with fewer losses of plants due to Fusarium wilt.