Application of PCR-Based Methods for Rapid Detection of Corn Ingredients in Processed Foods

Gabriadze, Inga; Kutateladze, Tamara; Vishnepolsky, Boris; Karseladze, Marina; Datukishvili, Nelly

doi:10.11648/j.ijnfs.20140303.21

Cited by 11 publications

(4 citation statements)

References 13 publications

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“…The PCR primers used in this study are shown in Table 1 . The species-specific primer pairs targeting soybean lectin gene and maize zein gene were taken from our previous publications (Kutateladze et al, 2013 ; Gabriadze et al, 2014 ). The published GenBank databases and DNA sequences were screened for the identification of GMO-specific DNA sequences and PCR primers.…”

Section: Methodsmentioning

confidence: 99%

New multiplex PCR methods for rapid screening of genetically modified organisms in foods

Datukishvili

Kutateladze²,

Gabriadze³

et al. 2015

Front. Microbiol.

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We present novel multiplex PCR methods for rapid and reliable screening of genetically modified organisms (GMOs). New designed PCR primers targeting four frequently used GMO specific sequences permitted identification of new DNA markers, in particular 141 bp fragment of cauliflower mosaic virus (CaMV) 35S promoter, 224 bp fragment of Agrobacterium tumefaciens nopaline synthase (NOS) terminator, 256 bp fragment of 5-enolppyruvylshikimate-phosphate synthase (epsps) gene and 258 bp fragment of Cry1Ab delta-endotoxin (cry1Ab) gene for GMO screening. The certified reference materials containing Roundup Ready soybean (RRS) and maize MON 810 were applied for the development and optimization of uniplex and multiplex PCR systems. Evaluation of amplification products by agarose gel electrophoresis using negative and positive controls confirmed high specificity and sensitivity at 0.1% GMO for both RRS and MON 810. The fourplex PCR was developed and optimized that allows simultaneous detection of three common transgenic elements, such as: CaMV 35S promoter, NOS terminator, epsps gene together with soybean-specific lectin gene. The triplex PCR developed enables simultaneous identification of transgenic elements, such as: 35S promoter and cry1Ab gene together with maize zein gene. The analysis of different processed foods demonstrated that multiplex PCR methods developed in this study are useful for accurate and fast screening of GM food products.

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Section: Methodsmentioning

confidence: 99%

New multiplex PCR methods for rapid screening of genetically modified organisms in foods

Datukishvili

Kutateladze²,

Gabriadze³

et al. 2015

Front. Microbiol.

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“…Ten healthy japonica seeds of each sample were randomly selected and cultivated at a room temperature of 25°. The fresh young leaves of the 3-leaf stage seedlings were mixed together, and the genomic DNA was extracted from 100 mg of each sample by the improved CTAB method 41 , 42 The sample was evenly mixed with 500 µL of CTAB buffer solution and 20 µL of proteinase K (20 mg/mL) evenly. After incubation at 65 °C for 60 min, 20 µL RNase A was added before centrifugation (1600 g for 10 min).…”

Section: Methodsmentioning

confidence: 99%

Exploring japonica rice epigenetic diversity in the main production regions of Heilongjiang Province

Zhang

et al. 2022

Sci Rep

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As a major epigenetic modification, DNA methylation plays an important role in coordinating plant responses to environmental changes. Methylation-sensitive amplified polymorphism (MSAP) technology was used in this study to investigate the epigenetic diversity of fifty japonica rice samples from five regions in Heilongjiang Province, China. In addition, the phenotypic indicators of japonica rice samples and the environmental conditions of the sampling sites were investigated and analysed. Based on the MSAP analysis technique, using eight pairs of selective primers, we identified a total of 551 amplified loci, of which 267 (48.5%) were classified as methylation loci. The methylation status and levels of the japonica rice genome in different regions differed significantly (p < 0.05). The results of the analysis of molecular variance (AMOVA) revealed that most of the molecular variation (91%) came from within the groups (regions) and was caused by individual variation within the region. Furthermore, the results of principal coordinates analysis (PCoA), cluster analysis, and population structure analysis indicated that there was no obvious correlation between the epigenetic differences and geographical locations, which may have been due to the limited range of sampling sites. When environmental factors, phenotypic indicators, and epigenetic data analysis are combined, it is easy to conclude that japonica rice grown in the same latitudinal region has increased epigenetic and phenotypic similarities due to similar climatic conditions and production practices.

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“…For the culture-independent assessment of OMF, DNA was initially extracted using a CTAB-based method [41]. Subsequently, amplicons were obtained using two pairs of primers (5.8S-OF/ITS4OF and 5.8S-OF/ITS4TUL, as described in [24]) that cover the entire diversity of rhizoctonian OMFs [42].…”

Section: Culture-dependent and -Independent Assessment Of Omfmentioning

confidence: 99%

Diversity of Mycorrhizal Fungi in Temperate Orchid Species: Comparison of Culture-Dependent and Culture-Independent Methods

Mennicken,

Paula,

Vogt-Schilb

et al. 2024

JoF

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Many orchid species are endangered due to anthropogenic pressures such as habitat destruction and overharvesting, meanwhile, all orchids rely on orchid mycorrhizal fungi (OMF) for seed germination and seedling growth. Therefore, a better understanding of this intimate association is crucial for orchid conservation. Isolation and identification of OMF remain challenging as many fungi are unculturable. In our study, we tested the efficiency of both culture-dependent and culture-independent methods to describe OMF diversity in multiple temperate orchids and assessed any phylogenetic patterns in cultivability. The culture-dependent method involved the cultivation and identification of single pelotons (intracellular hyphal coils), while the culture-independent method used next-generation sequencing (NGS) to identify root-associated fungal communities. We found that most orchid species were associated with multiple fungi, and the orchid host had a greater impact than locality on the variability in fungal communities. The culture-independent method revealed greater fungal diversity than the culture-dependent one, but despite the lower detection, the isolated fungal strains were the most abundant OMF in adult roots. Additionally, the abundance of NGS reads of cultured OTUs was correlated with the extent of mycorrhizal root colonization in orchid plants. Finally, this limited-scale study tentatively suggests that the cultivability character of OMF may be randomly distributed along the phylogenetic trees of the rhizoctonian families.

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Application of PCR-Based Methods for Rapid Detection of Corn Ingredients in Processed Foods

Cited by 11 publications

References 13 publications

New multiplex PCR methods for rapid screening of genetically modified organisms in foods

New multiplex PCR methods for rapid screening of genetically modified organisms in foods

Exploring japonica rice epigenetic diversity in the main production regions of Heilongjiang Province

Diversity of Mycorrhizal Fungi in Temperate Orchid Species: Comparison of Culture-Dependent and Culture-Independent Methods

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