Fusarium verticillioides and Aspergillus flavus cause Fusarium ear rot (FER) and Aspergillus ear rot (AER) of maize, respectively. Both pathogens are of concern to producers as they reduce grain yield and affect quality. F. verticillioides and A. flavus also contaminate maize grain with the mycotoxins fumonisins and aflatoxins, respectively, which has been associated with mycotoxicosis in humans and animals. The occurrence of common resistance mechanisms to FER and AER has been reported. Hence, ten Kenyan inbred lines resistant to AER and aflatoxin accumulation were evaluated for resistance to FER, F. verticillioides colonisation and fumonisin accumulation; and compared to nine South African lines resistant to FER and fumonisin accumulation. Field trials were conducted at three localities in South Africa and two localities in Kenya. FER severity was determined by visual assessment, while F. verticillioides colonisation and fumonisin content were quantified by real-time PCR and liquid chromatography tandem mass spectrometry, respectively. Significant genotype x environment interactions was determined at each location (P B 0.05). Kenyan inbred CML495 was most resistant to FER and F. verticillioides colonisation, and accumulated the lowest concentration of fumonisins across localities. It was, however, not significantly more resistant than Kenyan lines CML264 and CKL05015, and the South African line RO549 W, which also exhibited low FER severity (B5%), fungal target DNA (B0.025 ng lL -1 ) and fumonisin levels (B2.5 mg kg -1 ). Inbred lines resistant to AER and aflatoxin accumulation appear to be promising sources of resistance to F. verticillioides and fumonisin contamination.
Aspergillus flavus, causal agent of the Aspergillus ear rot (AER) of maize, also produces aflatoxins that cause aflatoxicosis in humans and livestock. Ten maize inbred lines were evaluated in replicated trials in two aflatoxicosis outbreak hot spots in Kenya and in three maize-growing areas in South Africa for resistance to AER, A. flavus colonization, and pre-harvest aflatoxin accumulation during the 2012/2013 growing season. AER severity was measured by visual assessment, while A. flavus colonization and aflatoxin content were quantified by real-time polymerase chain reaction (PCR) and liquid chromatography tandem mass spectrometry, respectively. Genotype by environment interaction (GEI) was determined using analysis of variance (ANOVA), additive main effects and multiplicative models (AMMI), and genotype plus by environment (GGE) biplot analyses. Stability of genotypes was evaluated using AMMI analysis. AER severity and fungal colonization significantly (p < 0.001) varied between genotypes. GEI influenced the severity of AER symptoms and aflatoxin accumulation significantly (p < 0.001), while fungal colonization was not affected. The inbred lines response was consistent for this trait in the test environments and was thus considered a desirable measure to indicate maize lines with a high risk of aflatoxin accumulation. CML495, CKL05019, LaPosta, and MIRTC5 were the least diseased lines, with the lowest aflatoxin contamination and a stable phenotypic response across the environments. Kiboko was determined as the ideal representative test environment, with discriminative ability of the genotypes for selection of the desired stable responses of the three traits.
Fusarium verticillioides causes Fusarium ear rot (FER) in maize (Zea mays L.), thus reducing grain quality, yield, and contaminates grains with fumonisins. Grain infection by these fungi occurs before harvest and selection of parental lines resistant to fumonisin accumulation for breeding purposes is the most effective and environmentally friendly control strategy for F. verticillioides. This study intended to evaluate F1 hybrids and F2 breeding populations in Kenya for improved resistance to FER and fumonisin contamination. Trials were artificially inoculated and FER severity, F. verticillioides accumulation, and fumonisin contamination were determined. Inheritance of resistance was also determined in the F1 hybrids. CML444 × MIRTC5, R119W × CKL05015, and CML444 × CKL05015 exhibited little to no FER and had the least fungal and fumonisin contamination, respectively. Inbred lines CML495, CKL05015, and P502 had negative, significant general combining ability (GCA) estimates for F. verticillioides colonization and fumonisin contamination, but positive, significant GCA estimates for 1,000‐kernel weight, respectively. The genotype × environment interaction was the main source of variation observed in the F2 populations with R119W × CKL05015 and CML444 × CKL05015 being the most tolerant to fungal and fumonisin contamination in Kiboko and MIRTC5 × CML495 the most tolerant in Katumani.
In this study, the possibility of sexual reproduction in sampled Aspergillus flavus strains was evaluated by assessing the distribution of mating type (MAT) genes, which are known to control sexual character among fungi, for two counties in Kenya. Forty-four isolates from Nandi and Makueni counties were genotyped by MAT using a multiplex polymerase chain reaction assay. The primer pair for the MAT1-1 amplified a 396 base pair (bp) fragment containing an α-box motif, and MAT1-2 primers targeted a 270 bp segment with a high mobility group protein. The MAT1-2 genes dominated in both regions although the frequency was higher in Nandi (75%) than in Makueni (54.17%). There were no MAT1-1 genes sampled in Nandi, and in Makueni their proportion was 15.91%. The percentage of isolates that amplified for both MAT genes in Makueni was 9.09%, while in Nandi it was 11.36%. Currently, use of aggressive aflatoxin non-producing A. flavus strains as biocontrol is the most promising preharvest aflatoxin control strategy in Kenya. However, we address the possibility of introduced biocontrol strains to breed with existing aflatoxin producing strains in nature, which could lead to the generation of A. flavus offspring capable of aflatoxin production while also being aggressive colonizers and possibly increasing the burden of aflatoxin exposure in food.
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