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Pathogenicity of the entomopathogenic fungus, Lecanicillium muscarium, against the sweetpotato whitefly Bemisia tabaci under laboratory and glasshouse conditions.

The potential for using the entomopathogenic fungus Lecanicillium muscarium to control the sweetpotato whitefly, Bemisia tabaci has been established in the laboratory by other studies. Laboratory studies however frequently overestimate the level of control achieved by biological control agents in the glasshouse. Before full-scale commercial or field development is considered, glasshouse trials are required to confirm laboratory results. Under both controlled laboratory and glasshouse conditions high mortality of second instar B. tabaci was recorded after application of L. muscarium. The potential of incorporating L. muscarium into integrated pest management strategies for the control of B. tabaci is discussed.

Animals↗

Enhancement of population size of a biological control agent and efficacy in control of bacterial speck of tomato through salicylate and ammonium sulfate amendments.

Sodium salicylate and ammonium sulfate were applied to leaf surfaces along with suspensions of the biological control agents Pseudomonas syringae Cit7(pNAH7), which catabolizes salicylate, and Cit7, which does not catabolize salicylate, to determine whether enhanced biological control of bacterial speck of tomato could be achieved. Foliar amendment with salicylate alone significantly enhanced the population size and the efficacy of Cit7(pNAH7), but not of Cit7, on tomato leaves. Application of ammonium sulfate alone did not result in enhanced population size or biological control efficacy of either Cit7(pNAH7) or Cit7; however, when foliar amendments with both sodium salicylate and ammonium sulfate were applied, a trend toward further increases in population size and biological control efficacy of Cit7(pNAH7) was observed. This study demonstrates the potential of using a selective carbon source to improve the efficacy of a bacterial biological control agent in the control of a bacterial plant disease and supports previous conclusions that the growth of P. syringae in the phyllosphere is primarily carbon limited and secondarily nitrogen limited.

Ammonium Sulfate↗

[Endophytic fungi in rose (Rosa hybrida) in Bogota, Colombia].

We have investigated the presence of endophytic fungi associated with rose plants (Rosa hybrida) in Colombia. Endophytic fungi were isolated from healthy leaves of ten ornamental roses plants from gardens cultured in malt extract, peptone, yeast extract agar plates (MPY). We sampled 560 leaves fragments, 56 per sample. Endophytic fungi comprised 92 isolates (16.4%); of these isolates, 41 were classified as sterile mycelium (without reproductive structures that allowed their identification), 31 isolates were identified to genus or to species, and 20 isolates could not be identified at all. The identified endophytic fungi were as follow: Nigrospora oryzae, Aureobasidium spp, Acremonium spp. The fungi Nodulisporium sp, Gliocladium virens, Cladosporium sp, Alternaria sp, Phoma sp and Chaetomium globosum were represented by one isolate each. Since the endophytic fungi are known for their capacity to produce metabolites with biological activity, it is possible that the microorganisms found in this study have potential as antagonist of rose pathogens.

Colombia↗

Effects of spring imidacloprid application for white grub control on parasitism of Japanese beetle (Coleoptera: Scarabaeidae) by Tiphia vernalis (Hymenoptera: Tiphiidae).

Imidacloprid, a relatively long residual neonicotinoid soil insecticide, is often applied to lawns and golf courses in spring for preventive control of root-feeding white grubs. We evaluated effects of such applications on spring parasitism of the overwintered third-instar Japanese beetle, Popillia japonica Newman, by Tiphia vernalis Rohwer, an introduced solitary ectoparasitoid. Natural rates of parasitism on a golf course rough were significantly lower in plots treated with full or one-half label rates of imidacloprid in early May compared with untreated turf. Parasitism also was reduced when female T. vernalis were exposed to imidacloprid residues on turf cores in the laboratory. Such exposures did not affect wasp mortality, longevity, survival, or developmental period of Tiphia larvae feeding on hosts in treated turf. They did, however, reduce wasps' ability to parasitize hosts in nontreated soil for at least 1-2 wk postexposure. In Y-trail choice tests, wasps that previously had been exposed to treated turf failed to respond normally to host frass trails in the soil. Female wasps did not avoid imidacloprid residues, imidacloprid-treated host frass, or host grubs that had previously been exposed to treated soil. This study indicates that applying imidacloprid in early spring can interfere with biological control by T. vernalis, whereas postponing preventive grub treatments until June or July, after the wasps' flight period, will help to conserve T. vernalis populations.

Animals↗

Aflatoxin reduction in corn through field application of competitive fungi.

Soil in corn plots was inoculated with nonaflatoxigenic strains of Aspergillus flavus and A. parasiticus during crop years 1994 to 1997 to determine the effect of application of the nontoxigenic strains on preharvest aflatoxin contamination of corn. Corn plots in a separate part of the field were not inoculated and served as controls. Inoculation resulted in significant increases in the total A. flavus/parasiticus soil population in treated plots, and that population was dominated by the applied strain of A. parasiticus (NRRL 21369). In the years when weather conditions favored aflatoxin contamination (1996 and 1997), corn was predominately colonized by A. flavus as opposed to A. parasiticus. In 1996, colonization by wild-type A. flavus was significantly reduced in treated plots compared with control plots, but total A. flavus/parasiticus colonization was not different between the two groups. A change to a more aggressive strain of A. flavus (NRRL 21882) as part of the biocontrol inoculum in 1997 resulted in a significantly (P < 0.001) higher colonization of corn by the applied strain. Weather conditions did not favor aflatoxin contamination in 1994 and 1995. In 1996, the aflatoxin concentration in corn from treated plots averaged 24.0 ppb, a reduction of 87% compared with the aflatoxin in control plots that averaged 188.4 ppb. In 1997, aflatoxin was reduced by 66% in treated corn (29.8 ppb) compared with control corn (87.5 ppb). Together, the data indicated that although the applied strain of A. parasiticus dominated in the soil, the nonaflatoxigenic strains of A. flavus were more responsible for the observed reductions in aflatoxin contamination. Inclusion of a nonaflatoxigenic strain of A. parasiticus in a biological control formulation for aflatoxin contamination may not be as important for airborne crops, such as corn, as for soilborne crops, such as peanuts.

Aflatoxins↗

In vitro studies on the potential for biological control on Aspergillus section Flavi by Kluyveromyces spp.

AIMS: Antagonist activity of Kluyveromyces spp. isolates on Aspergillus section Flavi was studied. METHODS AND RESULTS: The screening of isolates were made through studies of growth at different water activities and temperatures, index of dominance (I(D)), ecological similarity, antifungal activity and impact on aflatoxin B1 accumulation. High optical density was obtained at 25 and 30 degrees C and 48 h of incubation. Cell growth decreases with decrease in water activity. The predominant interaction was mutual intermingling at a(w) = 0.982 and 0.955, while at a(w) = 0.999 and 0.937 mutual inhibition for contact was exhibited. All isolates were catabolically identical to Aspergillus section Flavi and compete by nutritional source. At high water activities yeasts showed inhibitory activity on Aspergillus strains, inhibition percentages varied between 75 and 100%. The isolates Y9, Y14, Y16, Y22, Y25 and Y33 showed antifungal activity and inhibitory activity on aflatoxin B1 accumulation at all water activities assayed from all Aspergillus section Flavi strains. CONCLUSIONS: The data show that the isolates selected in a wide range of environmental conditions could exert their roll like biological control agents for Aspergillus section Flavi in storage maize ecosystem. SIGNIFICANCE AND IMPACT OF THE STUDY: Isolates of Kluyveromyces spp. may have practical value in the postharvest control of storage maize.

Aflatoxin B1↗

State-dependent impulsive models of integrated pest management (IPM) strategies and their dynamic consequences.

A state-dependent impulsive model is proposed for integrated pest management (IPM). IPM involves combining biological, mechanical, and chemical tactics to reduce pest numbers to tolerable levels after a pest population has reached its economic threshold (ET). The complete expression of an orbitally asymptotically stable periodic solution to the model with a maximum value no larger than the given ET is presented, the existence of which implies that pests can be controlled at or below their ET levels. We also prove that there is no periodic solution with order larger than or equal to three, except for one special case, by using the properties of the LambertW function and Poincare map. Moreover, we show that the existence of an order two periodic solution implies the existence of an order one periodic solution. Various positive invariant sets and attractors of this impulsive semi-dynamical system are described and discussed. In particular, several horseshoe-like attractors, whose interiors can simultaneously contain stable order 1 periodic solutions and order 2 periodic solutions, are found and the interior structure of the horseshoe-like attractors is discussed. Finally, the largest invariant set and the sufficient conditions which guarantee the global orbital and asymptotic stability of the order 1 periodic solution in the meaningful domain for the system are given using the Lyapunov function. Our results show that, in theory, a pest can be controlled such that its population size is no larger than its ET by applying effects impulsively once, twice, or at most, a finite number of times, or according to a periodic regime. Moreover, our theoretical work suggests how IPM strategies could be used to alter the levels of the ET in the farmers' favour.

Animals↗

Control of Culex quinquefasciatus in Myanmar (Burma) and India: 1960-1990.

During the past 30 years three major programmes of research have been carried out in Myanmar (Burma) and India on the ecology and control of Culex quinquefasciatus. To differing degrees the programmes have employed environmental, chemical, biological and genetic control strategies, but none has been an unqualified success. Results have been good when well-trained staff with substantial resources were employed, but much poorer when less-well-equipped general health workers took over the programmes. Overall, it would seem most cost-effective for health workers to concentrate their efforts on the elimination of the limited number of larval habitat categories which commonly contribute a large fraction of the adult population. Insecticides and biological control agents should serve as supplements, not as alternatives, to environmental management. Genetic control strategies have currently no place in C. quinquefasciatus programmes.

Animals↗

A simple model for predicting the effect of hygrothermal conditions on populations of house dust mite Dermatophagoides pteronyssinus (Acari: Pyroglyphidae).

A simple mite population index (MPI) model is presented which predicts the effect on house dust mite populations of any combination of temperature and relative humidity (RH). For each combination, the output is an index, or multiplication factor, such that 1.1 indicates 10% population growth and 0.9 indicates 10% population decline. To provide data for the model, laboratory experiments have been carried out using lab cultures of Dermatophagoides pteronyssinus. The population change was observed for mites held in steady-state conditions at different combinations of temperature and RH over 21 days. From the results, a best-fit equation has been derived which forms the basis of the MPI model. The results also enable a new term to be defined: the Population Equilibrium Humidity, PEH, the RH for a given temperature at which house dust mite populations neither grow nor decline. It is similar to Critical Equilibrium Humidity, the RH below which house dust mites are unable to maintain water balance, but relates to a population of mites (rather than a physiological phenomenon) and is more able to take account of the observed effects of extremes of temperature and RH. Compared with previous population models, the MPI model is potentially more accurate and comprehensive. It can be combined with other simple models (described in previous papers), such as BED, which simulates the average hygrothermal conditions in a bed, given room conditions, and Condensation Targeter II, which simulates room conditions given a range of easily obtainable inputs for climate, house type and occupant characteristics. In this way it is now possible, for any individual dwelling, to assess the most effective means of controlling mite populations by environmental means, such as by improving standards of ventilation and insulation, or by modifying the occupant behaviour that affects the hygrothermal environment within a dwelling. Although the MPI model requires further development and validation, it has already proved useful for understanding more clearly how the different hygrothermal conditions found in beds and bedrooms can affect mite populations. It has also demonstrated that there is considerable scope for controlling mites by environmental means in cold winter climates such as the UK.

Animals↗

Evaluation of biological control of sheep parasites using Duddingtonia flagrans under commercial farming conditions on the island of Gotland, Sweden.

Field trials, conducted over 3 consecutive years, were aimed at assessing farmer opinions of the practicality and effectiveness of using Duddingtonia flagrans to control nematode parasites in their flocks on the Swedish island of Gotland. These trials were also monitored by intensive parasitological investigation. On Gotland, lambing occurs in spring, and around mid-summer (late June), ewes and lambs are moved to saved pastures due to pasture deterioration caused by dry conditions. Weaned lambs are then returned to original lambing pastures in early autumn for finishing. One farm (B) was used for 2001-2003 and a second farm (N) was also used in 2002 and 2003. On each farm, two flocks (each of 20 ewes + twin lambs) were managed separately, namely: fungus group which received a daily supplement + fungal spores from lambing until the summer move (6 weeks) and: control group which received supplement only. For Farm B, the numbers of lambs that were marketed prior to the end of the grazing season, were 13, 18, 19 for the fungus treatment whereas corresponding numbers for the control treatment were 8, 16 and 11 for years 2001, 2002 and 2003, respectively. Final weights of the remaining lambs at the end of each year were also consistently heavier, and the numbers of lambs retained for finishing during winter were less, on the fungus treatment compared with the control treatment. On Farm N, similar results were recorded, with more lambs marketed earlier in the fungus group (25 and 19) compared with the control (19 and 15) in 2002 and 2003, respectively. The weights of the remaining lambs at the end of the trial in 2003 showed a 4.5 kg weight gain advantage of the fungus group compared to the controls. Tracer tests during autumn 2001 on Farm B, showed that Teladorsagia circumcincta plus Trichostrongylus spp. levels were significantly less on the fungus treatment (P=0.018). The summer/autumn of 2002 was one of the driest on record for Gotland. This resulted in very low levels of infective larval availability. But on both farms, T. circumcincta numbers were less on the fungus than on the control paddocks (P=0.048 on Farm B). In 2003 very low numbers of infective larvae were recorded in the autumn tracers for both treatments on both farms. Both farmer co-operators were encouraged with these results and consider that biological control of nematode parasites in their flocks, using D. flagrans, is of practical value.

Animals↗

Macrocyclic trichothecenes are undetectable in kudzu (Pueraria montana) plants treated with a high-producing isolate of Myrothecium verrucaria.

Myrothecium verrucaria was found to be an effective pathogen against kudzu grown in the greenhouse and the field. M. verrucaria produced large amounts of macrocyclic trichothecenes when cultured on solid rice medium, including epiroridin E (16.8 mg/g crude extract), epiisororidin E (1 mg/g), roridin E (8.7 mg/g), roridin H (31.3 mg/g), trichoverrin A (0.6 mg/g), trichoverrin B (0.1 mg/g), verrucarin A (37.4 mg/g), and verrucarin J (2.2 mg/g). Most of these toxins were also isolated from M. verrucaria spores and mycelia grown on potato dextrose agar medium, including epiroridin E (32.3 mg/g), epiisororidin E (28.6 mg/g), roridin E (0 mg/g), roridin H (60 mg/g), trichoverrin A (1.3 mg/g), trichoverrin B (1.8 mg/g), verrucarin A (13.8 mg/g), and verrucarin J (131 mg/g). When M. verrucaria was cultured on liquid media, the numbers but not the amounts of toxins decreased. Only epiroridin E (28.3 mg/g), epiisororidin E (29.6 mg/g), verrucarin B (195 mg/g) and verrucarin J (52.6 mg/g) were measured when the fungus was cultured on cornsteep medium. On soyflour-cornmeal broth M. verrucaria produced several toxins, including epiroridin E (58.1 mg/g), epiisororidin E (5.8 mg/g), verrucarin B (29.9 mg/g) and verrucarin J (32 mg/g). In contrast, no macrocyclic trichothecenes were detected by HPLC analysis of plant tissues of kudzu, sicklepod, and soybean treated with aqueous suspensions of M. verrucaria spores formulated with a surfactant. Chloroform-methanol extracts of kudzu leaves and stems treated with M. verrucaria spores were less cytotoxic to four cultured mammalian cell lines than the corresponding extracts from control plants. Purified macrocyclic trichothecenes (verrucarin A and T-2 toxin) were very cytotoxic to the same cell lines (< or = 2 ng/ml). These results show that neither intact macrocyclic trichothecenes nor toxic metabolites could be detected in plant tissues after treatment with M. verrucaria spores. These results argue for both safety and efficacy for the use of M. verrucaria in biological control of kudzu and other noxious weeds, and support proceeding to animal feeding trials for further evaluation of safety.

Ascomycota↗

Importance of ambient saturation deficits in an epizootic of the fungus Neozygites floridana in cassava green mites (Mononychellus tanajoa).

The mite-pathogenic fungus Neozygites floridana Fisher (Entomophthorales: Neozygitaceae) is considered to have potential for the biological control of the cassava green mite, Mononychellus tanajoa (Bondar). However, its activity is sporadic and laboratory data suggest a strong dependence on night-time saturation deficits for transmission. We report on an epizootic of this fungus in a mite population in northeastern Brazil. During the epizootic, host populations appeared to he limited by a combination of the pathogen and a predatory mite Neoseiulus idaeus (Acari: Phytoseiidae). When temperatures increased, the epizootic finished and the host population began to grow. Abiotic conditions could not explain the variation in host mortality following pickup of infective propagules in this epizootic. However, night-time saturation did help to explain the variation in transmission from infective cadavers to newly killed hosts. This supports laboratory observations that horizontal transmission between hosts is determined mainly by saturation deficits, while the process of infection is little affected by abiotic conditions. A further field observation was the near-absence of resting spores in dead mites (ca. 0.1% of cadavers), suggesting that the pathogen population was unsuccessful in producing inoculum to infect future M. tanajoa populations. The implications are that this pathogen will only be effective as a biological control agent in periods of high relative humidity, and establishment in new areas may be limited by resting spore formation.

Animals↗

Variation in tolerance and virulence in the chestnut blight fungus-hypovirus interaction.

Chestnut blight, caused by the fungus Cryphonectria parasitica, has been effectively controlled with double-stranded RNA hypoviruses in Europe for over 40 years. The marked reduction in the virulence of C. parasitica by hypoviruses is a phenomenon known as hypovirulence. This virus-fungus pathosystem has become a model system for the study of biological control of fungi with viruses. We studied variation in tolerance to hypoviruses in fungal hosts and variation in virulence among virus isolates from a local population in Italy. Tolerance is defined as the relative fitness of a fungal individual when infected with hypoviruses (compared to being uninfected); virulence is defined for each hypovirus as the reduction in fitness of fungal hosts relative to virus-free hosts. Six hypovirus-infected isolates of C. parasitica were sampled from the population, and each hypovirus was transferred into six hypovirus-free recipient isolates. The resulting 36 hypovirus-fungus combinations were used to estimate genetic variation in tolerance to hypoviruses, in hypovirus virulence, and in virus-fungus interactions. Four phenotypes were evaluated for each virus-fungus combination to estimate relative fitness: (i) sporulation, i.e., the number of asexual spores (conidia) produced; (ii) canker area on field-inoculated chestnut trees, (iii) vertical transmission of hypoviruses into conidia, and (iv) conidial germination. Two-way analysis of variance (ANOVA) revealed significant interactions (P < 0.001) between viruses and fungal isolates for sporulation and canker area but not for conidial germination or transmission. One-way ANOVA among hypoviruses (within each fungal isolate) and among fungal isolates (within each hypovirus) revealed significant genetic variation (P < 0.01) in hypovirus virulence and fungal tolerance within several fungal isolates, and hypoviruses, respectively. These interactions and the significant genetic variation in several fitness characters indicate the potential for future evolution of these characters. However, biological control is unlikely to break down due to evolution of tolerance to hypoviruses in the fungus because the magnitudes of tolerance and interactions were relatively small.

Ascomycota↗

Phenotypic and genetic diversity of Bacillus thuringiensis strains isolated in India active against Spodoptera litura.

Bacillus thuringiensis strains isolated from different agroclimatic regions of India were found to harbor cry1 family genes. Of 831 strains 18 that were found to produce 130- and 68-kDa mol wt proteins in sodium dodecyl sulfate polyacry1amide gel electrophoresis were subjected to bioassay against second instar larvae of Spodoptera litura. According to the time response curve, while the highest toxic activity against S. litura was observed in PBT-782 with an LT50 of 25.46 h, strains PBT-372, PBT-574, PBT-801, and PBT-716 in descending order of merit had LT50 values of 36.81, 48.18, 50.35, and 73.53 h. The results of the field experiment testing the efficacy of different B. thuringiensis strains in controlling S. litura larvae infecting peanut plants showed that the chemical insecticide chlorpyriphos was the most effective in controlling S. litura throughout the study period. However, among B. thuringiensis strains, PBT-372 was superior. All the B. thuringiensis strains except PBT-689 were found to contain cry1Ac1-type gene. However, only nine strains contained cry1Aa1 gene. While cry1Ab1 was present only in PBT-372 and PBT-689, cry1Ca1 was present in PBT-574, PBT-688, PBT-689, and PBT-695. cry1Da1 was detected only in PBT-688 and PBT-692. None of the strains contained cry1Ba1 and cry1Ea1 genes. When polymerase chain reaction analysis using cry1Ca1 primer was performed, PBT-695 produced an unexpected 739-bp product, which showed 33% homology with cry1Ca1 gene between nucleotides 1819 and 2107. Our results indicated that among the field-collected B. thuringiensis strains, PBT-372 harbors multiple cry-type genes and could be employed for biological control of insects.

Animals↗

Genetic variation and founder effects in the parasitoid wasp, Diaeretiella rapae (M'intosh) (Hymenoptera: Braconidae: Aphidiidae), affecting its potential as a biological control agent.

The effects of recent colonization on the aphid parasitoid, Diaeretiella rapae (M'Intosh) (Hymenoptera: Braconidae: Aphidiidae), in Western Australia were investigated. When compared with populations from the Old World, the results of a microsatellite analysis show that the insects have low allelic length and low allele frequency variation, revealing that these individuals experienced a significant founder effect. Marked genetic differentiation between populations was also revealed, which has potentially important implications for host utilization in this species when introduced to a new geographical area(s). Low genetic variation and gene flow in a founder population could limit evolutionary potential in Australia, including the ability of a population to mount a response to newly introduced hosts, such as the Russian wheat aphid, Diuraphis noxia (Mordvilko). Although the actual importance of genetic diversity in the success of biological control agents is unclear, current theory concerning the potential impact of genetic bottlenecks on additive genetic variance is discussed.

Animals↗

The predatory capability of three nematophagous fungi in the control of Haemonchus contortus infective larvae in ovine faeces.

The effect of oral administration of three different nematode-trapping fungi, in aqueous suspension containing either Dactylaria sp. or Arthrobotrys oligospora conidia or Duddingtonia flagrans chlamydospores, on the number of Haemonchus contortus infective larvae in sheep faeces, was evaluated. The three selected species of fungi produce three-dimensional adhesive nets in the presence of nematodes. Sixteen Creole sheep were divided into four groups of four animals each. Groups 1 and 2 were orally drenched with a suspension containing 2x10(7) conidia of either A. oligospora or Dactylaria sp. Group 3, received a similar treatment, with D. flagrans chlamydospores, instead of conidia, being administered, at the same dose. Group 4 acted as control, without any fungi. Faecal samples were collected directly from the rectum of each sheep and faecal cultures were prepared and incubated at 15 and 21 days. Larvae were recovered from faecal cultures and counted. The highest reduction of the nematode population occurred in the D. flagrans group, reaching reductions of 96.3% and 91.4% in individual samplings in plates incubated for 15 and 21 days, respectively. Arthrobotrys oligospora showed moderate reductions in the faecal larval population, ranging between 25-64% at 15 days incubation. In general, Dactylaria sp., was less efficient in its trapping ability. Despite the inconsistent results with Dactylaria sp., reduction percentages of 73.4% and 80.7% were recorded in individual samplings during the first and second days, in plates incubated for 15 days. Duddingtonia flagrans, was shown to be a potential biological control agent of H. contortus infective larvae.

Animals↗

In vitro influence of bacterial mixtures on Fusarium verticillioides growth and fumonisin B1 production: effect of seeds treatment on maize root colonization.

AIMS: Enterobacter cloacae, Microbacterium oleovorans, Pseudomonas solanacearum and Bacillus subtilis were investigated in order to evaluate: (i) the inoculum size of two bacterial mixtures on Fusarium verticillioides growth and fumonisin B1 production in vitro at different water activities and (ii) the efficacy of a seed treatment with the best bacterial mixture on F. verticillioides root colonization in greenhouse studies. METHODS AND RESULTS: The influence of bacterial mixtures (1 = E. cloacae and M. oleovorans and 2 = P. solanacearum and B. subtilis) to antagonize 13 F. verticillioides strains at different inoculum concentrations (10(8), 10(9) and 10(10) cells ml(-1)) and water activities (0.937, 0.955 and 0.982 aW) were examined. Antibiosis, growth rate and fumonisin B1 production were determined. Bacterial mixture 1 proved to exert the most effective control. Seed treatment with mixture 1 at 10(8) cells ml(-1) had the best inhibitory effect on F. verticillioides root colonization. CONCLUSIONS: These results suggest that the combination E. cloacae and M. oleovorans has the potential for the biological control of F. verticillioides as a maize seed inoculant. SIGNIFICANCE AND IMPACT OF THE STUDY: The application of this knowledge contributes to prevent the vertical transmission of F. verticillioides.

Antibiosis↗