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Fulltext Mycotoxigenic fungal growth inhibition and multi-mycotoxin reduction of potential biological control agents indigenous to grain maize

Yazid SNE, Tajudin NI, Razman NAA, Selamat J, Ismail SI, Sanny M, et al.

Mycotoxin Res, 2023 Aug;39(3):177-192.
PMID: 37219742 DOI: 10.1007/s12550-023-00484-4

The present work investigated the potential of fungal species from grain maize farms in Malaysia as antagonists against the indigenous mycotoxigenic fungal species and their subsequent mycotoxin production. Dual-culture assay was conducted on grain maize agar (GMA) with 12 strains of potential fungal antagonists namely Bjerkandra adusta, Penicillium janthinellum, Schizophyllum commune, Trametes cubensis, Trichoderma asperelloides, Trichoderma asperellum, Trichoderma harzianum, and Trichoderma yunnanense against seven mycotoxigenic strains namely Aspergillus flavus, Aspergillus niger, Fusarium verticillioides, and Fusarium proliferatum producing aflatoxins, ochratoxin A, and fumonisins, respectively. Based on fungal growth inhibition, Trichoderma spp. showed the highest inhibitory activity (73-100% PIRG, Percentage Inhibition of Radial Growth; 28/0 ID, Index of Dominance) against the tested mycotoxigenic strains. Besides, B. adusta and Tra. cubensis showed inhibitory activity against some of the tested mycotoxigenic strains. All fungal antagonists showed varying degrees of mycotoxin reduction. Aflatoxin B1 produced by A. flavus was mainly reduced by P. janthinellum, Tra. cubensis, and B. adusta to 0 ng/g. Ochratoxin A produced by A. niger was mainly reduced by Tri. harzianum and Tri. asperellum to 0 ng/g. Fumonisin B1 and FB2 produced by F. verticillioides was mainly reduced by Tri. harzianum, Tri. asperelloides, and Tri. asperellum to 59.4 and 0 µg/g, respectively. Fumonisin B1 and FB2 produced by F. proliferatum were mainly reduced by Tri. asperelloides and Tri. harzianum to 244.2 and 0 µg/g, respectively. This is the first study that reports on the efficacy of Tri. asperelloides against FB1, FB2, and OTA, P. janthinellum against AFB1, and Tra. cubensis against AFB1.

Matched MeSH terms: Zea mays/microbiology
Fulltext Characterization of Fusarium species section Liseola by restriction analysis of the IGS region

Heng MH, Baharuddin S, Latiffah Z

Genet. Mol. Res., 2012;11(1):383-92.
PMID: 22370941 DOI: 10.4238/2012.February.16.4

Fusarium species section Liseola namely F. fujikuroi, F. proliferatum, F. andiyazi, F. verticillioides, and F. sacchari are well-known plant pathogens on rice, sugarcane and maize. In the present study, restriction analysis of the intergenic spacer regions (IGS) was used to characterize the five Fusarium species isolated from rice, sugarcane and maize collected from various locations in Peninsular Malaysia. From the analysis, and based on restriction patterns generated by the six restriction enzymes, Bsu151, BsuRI, EcoRI, Hin6I, HinfI, and MspI, 53 haplotypes were recorded among 74 isolates. HinfI showed the most variable restriction patterns (with 11 patterns), while EcoRI showed only three patterns. Although a high level of variation was observed, it was possible to characterize closely related species and isolates from different species. UPGMA cluster analysis showed that the isolates of Fusarium from the same species were grouped together regardless of the hosts. We conclude that restriction analysis of the IGS regions can be used to characterize Fusarium species section Liseola and to discriminate closely related species as well as to clarify their taxonomic position.

Matched MeSH terms: Zea mays/microbiology
Fulltext The effect of tillage systems on phosphorus distribution and forms in rhizosphere and non-rhizosphere soil under maize (Zea mays L.) in Northeast China

Xomphoutheb T, Jiao S, Guo X, Mabagala FS, Sui B, Wang H, et al.

Sci Rep, 2020 04 20;10(1):6574.
PMID: 32313140 DOI: 10.1038/s41598-020-63567-7

An appropriate tillage method must be implemented by maize growers to improve phosphorus dynamics in the soil in order to increase phosphorus uptake by plant. The objective of this study was to investigate the effects of tillage systems on phosphorus and its fractions in rhizosphere and non-rhizosphere soils under maize. An experimental field was established, with phosphate fertilizers applied to four treatment plots: continuous rotary tillage (CR), continuous no-tillage (CN), plowing-rotary tillage (PR), and plowing-no tillage (PN). Under the different tillage methods, the available P was increased in the non-rhizosphere region. However, the concentration of available P was reduced in the rhizosphere soil region. The soil available P decreased with the age of the crop until the maize reached physiological maturity. The non-rhizosphere region had 132.9%, 82.5%, 259.8%, and 148.4% more available P than the rhizosphere region under the CR, PR, CN, and PN treatments, respectively. The continuous no-tillage method (CN) improved the uptake of soil phosphate by maize. The concentrations of Ca2-P, Ca8-P, Fe-P, Al-P and O-P at the maturity stage were significantly lower than other seedling stages. However, there was no significant relationship between total P and the P fractions. Therefore, a continuous no-tillage method (CN) can be used by farmers to improve phosphorus availability for spring maize. Soil management practices minimizing soil disturbance can be used to impove phosphorus availability for maize roots, increase alkaline phosphatase activity in the rhizosphere soil and increase the abundance of different phosphorus fractions.

Matched MeSH terms: Zea mays/microbiology
Efficacy of fungal and bacterial antagonists for controlling growth, FUM1 gene expression and fumonisin B1 production by Fusarium verticillioides on maize cobs of different ripening stages

Samsudin NI, Rodriguez A, Medina A, Magan N

Int J Food Microbiol, 2017 Apr 04;246:72-79.
PMID: 28213318 DOI: 10.1016/j.ijfoodmicro.2017.02.004

This study was carried out to examine the efficacy of two biocontrol agents (Clonostachys rosea 016, BCA1; Gram-negative bacterium, BCA5) for control of FUM1 gene expression and fumonisin B1 (FB1) production by F. verticillioides FV1 on maize cobs of different ripening stages: R3, Milk (0.985 aw); R4, Dough (0.976 aw); R5, Dent (0.958 aw). Initially, temporal studies on FUM1 gene expression and FB1 production were performed on maize kernels for up to 14days. This revealed that day 10 was optimum for both parameters, and was used in the biocontrol studies. Maize cobs were inoculated with 50:50 mixtures of the pathogen:antagonist inoculum and incubated in environmental chambers to maintain the natural aw conditions for ten days at 25 and 30°C. The growth rates of F. verticillioides FV1, the relative expression of the FUM1 gene and FB1 production were quantified. It was found that, aw×temp had significant impacts on growth, FUM1 gene expression and FB1 production by F. verticillioides FV1 on maize cobs of different maturities. The fungal antagonist (BCA1) significantly reduced FB1 contamination on maize cobs by >70% at 25°C, and almost 60% at 30°C regardless of maize ripening stage. For the bacterial antagonist (BCA5) however, FB1 levels on maize cobs were significantly decreased only in some treatments. These results suggest that efficacy of antagonists to control mycotoxin production in ripening maize cobs needs to take account of the ecophysiology of the pathogen and the antagonists, as well as the physiological status of the maize during silking to ensure effective control.

Matched MeSH terms: Zea mays/microbiology*
Formulation of maize- and peanut-based semi-synthetic growth media for the ecophysiological studies of aflatoxigenic Aspergillus flavus in maize and peanut agro-ecosystems

Yazid SNE, Thanggavelu H, Mahror N, Selamat J, Samsudin NIP

Int J Food Microbiol, 2018 Oct 03;282:57-65.
PMID: 29913332 DOI: 10.1016/j.ijfoodmicro.2018.06.007

In studying the ecophysiology of fungal phytopathogens, several stages are involved (in vitro, greenhouse, in planta). Most in vitro studies extensively utilise the general growth media such as Potato Dextrose Agar and Malt Extract Agar. Although the crop components in these media serve as excellent carbon sources and yield luxuriant growth, they are not naturally contaminated with Aspergillus flavus and thus might result in under- or overestimation of its actual toxigenic potentials. Empirical data on the formulation of semi-synthetic growth medium mimicking the natural crop commonly contaminated by A. flavus for the ecophysiological studies in vitro are scarce. The present work was aimed at investigating the ecophysiology of A. flavus on commercial growth media (PDA, MEA); formulating maize- and peanut-based semi-synthetic growth media using two methods of raw material preparation (milling, hot water extraction) at different concentrations (1, 3, 5, 7, 9% w/v), and comparing the ecophysiological parameters between commercial and formulated growth media. Growth rates were obtained by computing the hyphal expansion data into y = mx + c equation. AFB1 was quantified using high performance liquid chromatography with fluorescence detector. Formulated media were found to yield significantly higher growth rates when compared to commercial media. However, commercial media yielded significantly higher AFB1 when compared to all formulated media. Between the two formulations, milling yielded significantly higher growth rates and AFB1 when compared to hot water extraction. Although in vitro data cannot directly extrapolate in planta performance, results obtained in the present work can be used to gauge the actual toxigenic potential of A. flavus in maize and peanut agro-ecosystems.

Matched MeSH terms: Zea mays/microbiology*
Fulltext Determination of the Use of Lactobacillus plantarum and Propionibacterium freudenreichii Application on Fermentation Profile and Chemical Composition of Corn Silage

Abdul Rahman N, Abd Halim MR, Mahawi N, Hasnudin H, Al-Obaidi JR, Abdullah N

Biomed Res Int, 2017;2017:2038062.
PMID: 28503566 DOI: 10.1155/2017/2038062

Corn was inoculated with Lactobacillus plantarum and Propionibacterium freudenreichii subsp. shermanii either independently or as a mixture at ensiling, in order to determine the effect of bacterial additives on corn silage quality. Grain corn was harvested at 32-37% of dry matter and ensiled in a 4 L laboratory silo. Forage was treated as follows: bacterial types: B0 (without bacteria-control), B1 (L. plantarum), B2 (P. freudenreichii subsp. shermanii), and B3 (combination of L. plantarum and P. freudenreichii subsp. shermanii). Each 2 kg of chopped forage was treated with 10 mL of bacterial culture and allowed to ferment for 27 days. The first experiment determined the most suitable wavelength for detection of bacteria (490 nm and 419 nm for B1 and B2, resp.) and the preferable inoculation size (1 × 105 cfu/g). The second experiment analysed the effect of B1 and B2 applied singly or as a mixture on the fermentation characteristics and quality of corn silage. L. plantarum alone increased crude protein (CP) and reduced pH rapidly. In a mixture with P. freudenreichii, the final pH was the lowest compared to other treatments. As a mixture, inclusion of bacteria resulted in silage with lower digestibility than control. Corn silage treated with L. plantarum or P. freudenreichii either alone or mixed together produced desirable silage properties; however, this was not significantly better than untreated silage.

Matched MeSH terms: Zea mays/microbiology
Prevalence of mycotoxigenic fungi and assessment of aflatoxin contamination: a multiple case study along the integrated corn-based poultry feed supply chain in Malaysia

Nasaruddin N, Jinap S, Samsudin NI, Kamarulzaman NH, Sanny M

J Sci Food Agric, 2021 Mar 30;101(5):1812-1821.
PMID: 32893877 DOI: 10.1002/jsfa.10795

BACKGROUND: Corn, a main feed ingredient in the livestock industry, is one of the most susceptible crops to fungal infection and aflatoxin contamination. Livestock feeding on aflatoxin (AF)-contaminated feed have been shown to experience feed refusal, and decreased growth rate, milk production, and feed efficiency. In poultry, AF poisoning causes weight loss, poor feed efficiency, and reduced egg production and egg weight. The present work therefore aimed to determine the prevalence of mycotoxigenic fungi and the occurrence of AF contamination along the integrated corn-based poultry feed supply chain in Malaysia. A total of 51 samples were collected from different points along the feed supply chain from integrated poultry feed companies. The samples were subjected to mycological analyses (fungal isolation, enumeration, identification), and AFs were quantified by high-performance liquid chromatography equipped with a fluorescence detector (HPLC-FLD).
RESULTS: Samples collected from sampling point 1 (company A) and sampling point 9 (company B) yielded the highest total fungal load (>log 4 CFU g-1 ). The prevalent fungal genera isolated were Aspergillus, Fusarium, and Penicillium spp. Aflatoxin B1 was detected in 8.3% of corn samples, and 7.4% of corn-based poultry feed samples along the feed supply chain, whereas AFs B2 , G1 , and G2 were not detected.
CONCLUSION: The incidence of mycotoxigenic fungi along the integrated poultry feed supply chain warrant continuous monitoring of mycotoxin contamination to reduce the exposure risk of mycotoxin intake in poultry. © 2020 Society of Chemical Industry.

Matched MeSH terms: Zea mays/microbiology
Fulltext Plant Growth-Promoting Rhizobacteria Inoculation to Enhance Vegetative Growth, Nitrogen Fixation and Nitrogen Remobilisation of Maize under Greenhouse Conditions

Kuan KB, Othman R, Abdul Rahim K, Shamsuddin ZH

PLoS One, 2016;11(3):e0152478.
PMID: 27011317 DOI: 10.1371/journal.pone.0152478

Plant growth-promoting rhizobacteria (PGPR) may provide a biological alternative to fix atmospheric N2 and delay N remobilisation in maize plant to increase crop yield, based on an understanding that plant-N remobilisation is directly correlated to its plant senescence. Thus, four PGPR strains were selected from a series of bacterial strains isolated from maize roots at two locations in Malaysia. The PGPR strains were screened in vitro for their biochemical plant growth-promoting (PGP) abilities and plant growth promotion assays. These strains were identified as Klebsiella sp. Br1, Klebsiella pneumoniae Fr1, Bacillus pumilus S1r1 and Acinetobacter sp. S3r2 and a reference strain used was Bacillus subtilis UPMB10. All the PGPR strains were tested positive for N2 fixation, phosphate solubilisation and auxin production by in vitro tests. In a greenhouse experiment with reduced fertiliser-N input (a third of recommended fertiliser-N rate), the N2 fixation abilities of PGPR in association with maize were determined by 15N isotope dilution technique at two harvests, namely, prior to anthesis (D50) and ear harvest (D65). The results indicated that dry biomass of top, root and ear, total N content and bacterial colonisations in non-rhizosphere, rhizosphere and endosphere of maize roots were influenced by PGPR inoculation. In particular, the plants inoculated with B. pumilus S1r1 generally outperformed those with the other treatments. They produced the highest N2 fixing capacity of 30.5% (262 mg N2 fixed plant-1) and 25.5% (304 mg N2 fixed plant-1) of the total N requirement of maize top at D50 and D65, respectively. N remobilisation and plant senescence in maize were delayed by PGPR inoculation, which is an indicative of greater grain production. This is indicated by significant interactions between PGPR strains and time of harvests for parameters on N uptake and at. % 15Ne of tassel. The phenomenon is also supported by the lower N content in tassels of maize treated with PGPR, namely, B. pumilus S1r1, K. pneumoniae Fr1, B. subtilis UPMB10 and Acinetobacter sp. S3r2 at D65 harvest. This study provides evidence that PGPR inoculation, namely, B. pumilus S1r1 can biologically fix atmospheric N2 and provide an alternative technique, besides plant breeding, to delay N remobilisation in maize plant for higher ear yield (up to 30.9%) with reduced fertiliser-N input.

Matched MeSH terms: Zea mays/microbiology*
Virulence, Host-Selective Toxin Production, and Development of Three Cochliobolus Phytopathogens Lacking the Sfp-Type 4'-Phosphopantetheinyl Transferase Ppt1

Zainudin NA, Condon B, De Bruyne L, Van Poucke C, Bi Q, Li W, et al.

Mol Plant Microbe Interact, 2015 Oct;28(10):1130-41.
PMID: 26168137 DOI: 10.1094/MPMI-03-15-0068-R

The Sfp-type 4'-phosphopantetheinyl transferase Ppt1 is required for activation of nonribosomal peptide synthetases, including α-aminoadipate reductase (AAR) for lysine biosynthesis and polyketide synthases, enzymes that biosynthesize peptide and polyketide secondary metabolites, respectively. Deletion of the PPT1 gene, from the maize pathogen Cochliobolus heterostrophus and the rice pathogen Cochliobolus miyabeanus, yielded strains that were significantly reduced in virulence to their hosts. In addition, ppt1 mutants of C. heterostrophus race T and Cochliobolus victoriae were unable to biosynthesize the host-selective toxins (HST) T-toxin and victorin, respectively, as judged by bioassays. Interestingly, ppt1 mutants of C. miyabeanus were shown to produce tenfold higher levels of the sesterterpene-type non-HST ophiobolin A, as compared with the wild-type strain. The ppt1 strains of all species were also reduced in tolerance to oxidative stress and iron depletion; both phenotypes are associated with inability to produce extracellular siderophores biosynthesized by the nonribosomal peptide synthetase Nps6. Colony surfaces were hydrophilic, a trait previously associated with absence of C. heterostrophus Nps4. Mutants were decreased in asexual sporulation and C. heterostrophus strains were female-sterile in sexual crosses; the latter phenotype was observed previously with mutants lacking Nps2, which produces an intracellular siderophore. As expected, mutants were albino, since they cannot produce the polyketide melanin and were auxotrophic for lysine because they lack an AAR.

Matched MeSH terms: Zea mays/microbiology*