Inoculation methods and agressiveness of Macrophomina phaseolina isolates in cowpea

ABSTRACT: The fungi Macrophomina phaseolina is the charcoal rot causal agent, one of the most important cowpea crop disease in semiarid regions can causes 100% yield losses. The search for resistant genotypes requires efficient phenotyping. In addition, there is the problem of great variation in aggressiveness between isolates. This study aimed to 1) test three methods of inoculation in semiarid conditions, and 2) to evaluate the aggressiveness of isolates of M. phaseolina. In the first experiment carried out in greenhouse, the inoculations methods were evaluated, using two cowpea lines, three inoculation methods and three pathogen isolates. On the second experiment, fifteen M. phaseolina isolates were inoculated in one cultivar to evaluate their aggressiveness. By assessing the length of the lesions and the severity of the disease using an index, we identified the toothpick inoculation method as the most efficient. Toothpick method allowed to discriminate the genotypes and the aggressiveness of the pathogen.

Effects of Trichoderma strigosellum in Eucalyptus urophylla Development and Leaf-Cutting Ant Behavior

Fungal endophytes can protect plants against herbivory and be used to control leaf-cutting ants. In this study, we aimed to evaluate the potential of endophytic colonization of Eucalyptus urophylla by three filamentous fungal species and their influence on the plant development and foraging behavior of Atta sexdens. The study design was completely randomized and comprised a factorial scheme of 4 × 3, three antagonistic fungal species (Escovopsis sp., Metarhizium anisopliae, and Trichoderma strigosellum) of the leaf-cutting ant, and one control and three inoculation methods (conidial suspension via foliar spray [FS] and soil drench [SD] inoculation, and seedlings inoculated with mycelium [SWM]). The SWM method allowed T. strigosellum to colonize all plant organs, and these plants exhibited higher height, leaf number, shoot dry mass, and total dry mass than the ones subjected to the other inoculation methods. The SWM method increased the plant height than the control plants and those inoculated with Escovopsis sp. and M. anisopliae. Trichoderma strigosellum, previously isolated from soil, colonized E. urophylla plants and positively influenced their development, as demonstrated by the SWM method. Trichoderma strigosellum promoted the increase in E. urophylla height compared with when the FS and SD methods were used (by 19.62% and 18.52%, respectively). Our results reveal that A. sexdens workers preferentially began cutting the leaves from plants not previously colonized by T. strigosellum. This behavior can be explained by modifications in the phenotypic traits of the eucalyptus leaves.

Assessment of Inoculation Methods of Thielaviopsis paradoxa (De Seynes) Höhn into Oil Palm Seedlings under Greenhouse Conditions

Oil palm (Elaeis guineensis Jacq. and Elaeis Oleifera Cortes) is one of the most important oil crops in the world. Colombia is the fourth-largest oil palm producer worldwide. However, oil palm diseases are a significant factor affecting yield. Thielaviopsis paradoxa (De Seynes) Höhn is a pathogen that affects young palm trees, causing spear rot. Four disease establishment methods were studied to replicate, in a controlled environment, the symptoms of the disease found in the field. Young palm trees were inoculated with a suspension of endoconidia using either local infiltration, drip, scissor cut, or direct contact with agar blocks bearing mycelia and conidia. The effects of the inoculation methods were studied in dose-method-disease severity experiments conducted in a greenhouse under controlled conditions. All four methods resulted in T. paradoxa infections and the development of symptoms of the disease. The disease severity was correlated with the method and dose of inoculation. In trials to test Koch’s postulates, T. paradoxa was isolated from areas of disease progression in the inoculated trees, but the teleomorph Ceratocystis paradoxa (Dade) Moreau was not observed. A photographic record of the infection process at different times post-infection was compiled. Given that establishing the disease through artificial inoculation is essential for assessing plant pathogenesis, this study determined that the local infiltration method (1 × 106 endoconidia mL−1) and a 3–7 day incubation period were critical for the development of symptoms as severe as those observed in natural infections in the field.

Morphometry and production of maize inoculated with Azospirillum brasilense

This study aimed to analyze the influence of inoculation methods with Azospirillum brasilense in morphometric and nutritional parameters of the maize crop. Maize plants, hybrid Formula VT®, were grown under different forms of inoculation: absence of inoculation; seed inoculation; leaf inoculation; seed inoculation associated to leaf inoculation. In the phenological stages V8 and VT the parameters number of leaves, aerial height, root volume, stem diameter, besides the leaves, stem, sheath, root and total dry mass and the leaf content of nitrogen, phosphorus and potassium were determined. In the R6 stage, in addition to the previous evaluations the following parameters were determined: ear diameter and length, number of kernel rows, number of kernels per row, the reproductive structure and thousand grains dry mass, and total number of grains, besides the grain’s content of nitrogen, phosphorus, and potassium. The SPAD index from the leave’s apex, medial and basal thirds was measured fortnightly. The inoculation methods with A. brasilense had little influence in maize’s morphometric and nutritional parameters, also not influencing in production. Maize plants inoculated with A. brasilense, via seed and via seed associated to leaf spraying, positively stood out for the stem diameter, leaves dry mas, root volume, and for the nitrogen, phosphorus, and potassium leaf content.

Two Different Inoculation Methods Unveiled the Relative Independence of DON Accumulation in Wheat Kernels from Disease Severity on Spike after Infection by Fusarium Head Blight

Fusarium head blight (FHB) causes wheat yield loss and mycotoxin (deoxynivalenol, DON) accumulation in wheat kernel. Developing wheat cultivars with overall resistance to both FHB spread within a spike and DON accumulation in kernels is crucial for ensuring food security and food safety. Here, two relatively novel inoculation methods, bilateral floret inoculation (BFI) and basal rachis internode injection (BRII), were simultaneously employed to evaluate disease severity and DON content in kernels in a segregating population of recombinant inbred lines (RILs) developed from Ning 7840 (carrying Fhb1) and Clark (without Fhb1). Under both inoculation methods, four contrasting combinations of disease severity and DON content were identified: high severity/high DON (HSHD), high severity/low DON (HSLD), low severity/high DON (LSHD) and low severity/low DON (LSLD). Unexpectedly, the BRII method clearly indicated that disease severity was not necessarily relevant to DON concentration. The effects of Fhb1 on disease severity, and on DON concentrations, agreed very well across the two methods. Several lines carrying Fhb1 showed extremely higher severity and (or) DON content under both inoculation methods. The “Mahalanobis distance” (MD) method was used to rate overall resistance of a line by inclusion of both disease severity and DON content over both methods to select LSLD lines.

A New Identification Method Reveals the Resistance of an Extensive-Source Radish Collection to Plasmodiophora brassicae Race 4

Raphanus sativus, an important cruciferous vegetable, has been increasingly affected by clubroot disease. Establishing a stable and accurate resistance identification method for screening resistant germplasms is urgently needed in radish. In this study, the influence of inoculum concentration, inoculation methods, and pH of the substrate on disease occurrence was studied. The result showed that the disease index (DI) was highest at 2 × 108 spores/mL, the efficiency of two-stage combined inoculation methods was higher than others, and pH 6.5 was favorable for the infection of P. brassicae. By using this new method, DIs of 349 radish germplasms varying from 0.00 to 97.04, presented significantly different levels of resistance. Analysis showed that 85.06% germplasms from China were susceptible to P. brassicae, whilst 28 accessions were resistant and mainly distributed in east, southwest, northwest, and south-central China. Most of the exotic germplasms were resistant. Repeated experiments verified the stability and reliability of the method and the identity of germplasm resistance. In total, 13 immune, 5 highly resistant and 21 resistant radish accessions were identified. This study provides an original clubroot-tolerance evaluation technology and valuable materials for the development of broad-spectrum resistant varieties for sustainable clubroot management in radish and other cruciferous crops.

Successful Plant Growth-Promoting Microbes: Inoculation Methods and Abiotic Factors

Plant-microbe interactions have been the subject of several biotechnological studies, seeking sustainable development and environmental conservation. The inoculation of plant growth-promoting microbes (PGPM) in agricultural crops is considered an environmental-friendly alternative to chemical fertilization. Microbial inoculants are mainly inoculated onto seeds, roots and soil. PGPM improve plant growth by enhancing the availability of nutrients, the regulation of phytohormones, and by increasing plant tolerance against biotic and abiotic stresses. One of the main obstacles with PGPM research are the inconsistent results, which may be the result of inoculation methods and abiotic factors, such as soil (nutrient or heavy metal contents and pH), water availability, light intensity and temperature. This review addresses how the PGPM inoculants act on plant growth, what mechanisms they use to survive under stressful environmental conditions, and how inoculation methods and abiotic factors can interfere on the success of microbial inoculation in plants, serving as a basis for research on plants-microorganisms interaction.

Impact of Inoculation Methods and Fruit Maturity on Development of Fusarium Fruit Rot (Fusarium musae) Disease in Banana

Background: Fusarium fruit rot symptoms appear at all stages of banana fruit once infection initiates and progress from the stylar end as dark brown to black small spots which eventually turns black rotten areas leading to rotten pulp which renders the fruit unmarketable. Very meagre research work has been done on Fusarium fruit rot disease of banana in India and hence the present investigation on Fusarium fruit rot diseases of banana was undertaken south Gujarat condition. Methods: During the period 2018-2019 tested different inoculation methods were tested on the development of banana Fusarium fruit rot disease viz., stylar end pricking, rubbing, pricking at epicarp and without injury on banana fruit. Disease severity and incidence were recorded at 4th and 8th day after inoculation. In order to find out the most vulnerable stage of fruit for initiation of infection and development of Fusarium fruit rot disease, three stages of fruits were selected i.e., unripe, semi-ripe and ripe.Conclusion: Among, the different methods with fruit injury, stylar end pricking method was found best for the infection and development of Fusarium fruit rot (34.14%) after 8th days of inoculation followed by pricking at epicarp (27.79%). Further it was observed that injury of fruit surface is essential for infection and development of Fusarium fruit rot in banana. However, the severity of Fusarium fruit rot revealed that the highest severity was recorded in ripe fruits (30.21%) as compared to semi ripe (30.08%) and unripe (14.70%) fruits.