Prolamin Content and Grain Weight in RNAi Silenced Wheat Lines Under Different Conditions of Temperature and Nitrogen Availability

Marín-Sanz, Miriam; Giménez, María J.; Barro, Francisco; Savin, Roxana

doi:10.3389/fpls.2020.00314

Cited by 11 publications

(10 citation statements)

References 76 publications

(93 reference statements)

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“…Almost no relationship between the two parameters is further confirmed by the low negative correlation (r = −0.13). This is in conflict with the relationships known in the so-called Osborne fractionation of proteins; when in cereals, a close relationship is declared between the content of the crude protein and the storage prolamine fraction [ 25 ]. The non-specific reaction of avenins with homologous gluten epitopes has already been described by Londono et al [ 11 ].…”

Section: Resultsmentioning

confidence: 99%

“…Another hitherto unmentioned factor influencing the content variability of epitopes is also the influence of weather conditions (e.g., temperature stress), which affect the biosynthesis of various fractions of prolamins and thus the abundance of avenins containing immunoreactive gluten-like sequences [ 25 ]. The presumed cultivar-specific cross-reactivity is thus just one of the other factors mentioned above that affect the quantification of the gluten content in oats by current commercial kits (e.g., year and cropping system, kit detection limit, random interaction with non-specific avenin epitopes, inability to distinguish external gluten contamination from reaction with avenins, etc.).…”

Section: Resultsmentioning

confidence: 99%

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Specific Avenin Cross-Reactivity with G12 Antibody in a Wide Range of Current Oat Cultivars

Dvořāček

Kotrbová-Kozak

Kozová-Doležalová

et al. 2022

Foods

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Current clinical studies confirm that the consumption of oats for people suffering from celiac disease is safe. Some studies have confirmed different levels of immunoreactive gluten epitopes of oats in different cultivars, while others explain these differences due to contamination with gluten-rich species or as random cross-reactivity ELISA of homologous oat epitopes with anti-wheat gliadin antibodies. The aim of our two-year study was therefore to map cross-reactive oat epitopes in a set of 132 oat cultivars using a G12-based ELISA kit. The results were focused on the varietal and annual level of cross-reactivity (interference) of avenin epitopes with the G12 antibody on the identification of potential cultivars with significantly different interferences and assessing the degree of risk of possible false-contamination with external gluten. Although repeated evaluations confirmed high year-to-year variability (RSD ≥ 30%) in approximately 2/3 of the cultivars, the content of interfering avenin epitopes with G12 did not exceed the considered safe limit (20 mg·kg−1) for celiacs. At the same time, not only annual but, above all, significant cultivar dependences in the interference of avenins to the G12 antibody were demonstrated. Genetic dependence was further confirmed in connection with the proven avenin polymorphism as well as immunoblotting with the identification of interfering peptides with the G12 antibody in the 25 and 30 kDa regions. It was the occurrence of two bands around 30 kDa that predominantly occurred in oat cultivars with a relatively higher content of cross-reactive avenins (12–16 mg·kg−1). Due to the fact that the contents of interfering avenins ranged in several cultivars even over 16 mg·kg−1, the choice of a suitable oat cultivar may be crucial for gluten-free food producers, as it reduces the risk of a possible false-response of the commercial ELISA kits when checking the real-gluten contamination.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Specific Avenin Cross-Reactivity with G12 Antibody in a Wide Range of Current Oat Cultivars

Dvořāček

Kotrbová-Kozak

Kozová-Doležalová

et al. 2022

Foods

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“…Four wheat lines with gliadins down-regulated by RNAi have been characterized under heat stress and varying N availability, and silencing was found to be stable under heat stress [ 116 ]. Several temperature-responsive miRNA species have been identified in many plant species [ 117 ].…”

Section: Vision Two: Abiotic Stressmentioning

confidence: 99%

Tuning Beforehand: A Foresight on RNA Interference (RNAi) and In Vitro-Derived dsRNAs to Enhance Crop Resilience to Biotic and Abiotic Stresses

Abdellatef

Kamal

Tsujimoto

2021

IJMS

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Crop yield is severely affected by biotic and abiotic stresses. Plants adapt to these stresses mainly through gene expression reprogramming at the transcriptional and post-transcriptional levels. Recently, the exogenous application of double-stranded RNAs (dsRNAs) and RNA interference (RNAi) technology has emerged as a sustainable and publicly acceptable alternative to genetic transformation, hence, small RNAs (micro-RNAs and small interfering RNAs) have an important role in combating biotic and abiotic stresses in plants. RNAi limits the transcript level by either suppressing transcription (transcriptional gene silencing) or activating sequence-specific RNA degradation (post-transcriptional gene silencing). Using RNAi tools and their respective targets in abiotic stress responses in many crops is well documented. Many miRNAs families are reported in plant tolerance response or adaptation to drought, salinity, and temperature stresses. In biotic stress, the spray-induced gene silencing (SIGS) provides an intelligent method of using dsRNA as a trigger to silence target genes in pests and pathogens without producing side effects such as those caused by chemical pesticides. In this review, we focus on the potential of SIGS as the most recent application of RNAi in agriculture and point out the trends, challenges, and risks of production technologies. Additionally, we provide insights into the potential applications of exogenous RNAi against biotic stresses. We also review the current status of RNAi/miRNA tools and their respective targets on abiotic stress and the most common responsive miRNA families triggered by stress conditions in different crop species.

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“…The high inter-year variability in immunoreactive gluten content in the majority of studied oat genotypes can be explained by combination of two major factors. The first one represent environmental conditions such as temperature stress (extreme temperatures) which affect the biosynthesis of different prolamins fractions and thus the abundance of avenins containing immunoreactive gluten-like sequences [20]. The other factor is associated with an identification of homologous gluten-like epitopes in avenins which are slightly different from /not identical with wheat gluten epitopes [6].…”

Section: Contents and Variability Of Immunnoreactive Gluten Peptides In The Set Of Oat Cultivarsmentioning

confidence: 99%

Identification of Perspective Oat Cultivars with a Minimum Content of Gluten Homologous Peptides

Dvořāček¹,

Kotrbová-Kozak²,

Kozová-Doležalová³

et al. 2020

Preprint

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There is still an ambiguous opinion on oat regarding its safety for people with celiac disease (CD). Some studies have confirmed different content of oat immunoreactive epitopes in different cultivars while others explain the differences in consequence of cross-contamination with gluten-rich species or as ELISA cross-reactivity of oat homological epitopes with antibodies against wheat gliadin. Our study was based on a two-year mapping of oat immunoreactive epitopes in a set of 132 oat cultivars using the G12-based ELISA kit. Although repeated evaluation confirmed high interannual variability (RSD ≥ 30%) in approximately 2/3 of the cultivars, the permitted gluten content (20 mg kg-1) has not been exceeded except for contaminated cultivar Sirene. The polymorphism of purified avenins determined by SDS-PAGE revealed the occurrence of 2 bands around 30 kDa in oat cultivars with relatively high gluten content (12-16 mg kg-1) except for the cultivar Leo while this pattern occurred only in 50% oat cultivars with low gluten content. Quantification of gluten epitopes on purified avenin unit further revealed the three materials (Mojacar, Maris Oberon, SG-K 16370) with the lowest gluten content and presence of one band at 30 kDa. The band pattern at 30 kDa thus represents a promising breeding marker.

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Prolamin Content and Grain Weight in RNAi Silenced Wheat Lines Under Different Conditions of Temperature and Nitrogen Availability

Cited by 11 publications

References 76 publications

Specific Avenin Cross-Reactivity with G12 Antibody in a Wide Range of Current Oat Cultivars

Specific Avenin Cross-Reactivity with G12 Antibody in a Wide Range of Current Oat Cultivars

Tuning Beforehand: A Foresight on RNA Interference (RNAi) and In Vitro-Derived dsRNAs to Enhance Crop Resilience to Biotic and Abiotic Stresses

Identification of Perspective Oat Cultivars with a Minimum Content of Gluten Homologous Peptides

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