Linoleic Acid:Dihomo-γ-Linolenic Acid Ratio Predicts the Efficacy of Zn-Biofortified Wheat in Chicken (Gallus gallus)

The amount of Zn absorbed from Zn-biofortified wheat material has been determined using an <i>in vivo</i> model of Zn absorption. The erythrocyte linoleic:dihomo -γ-linolenic acid (LA:DGLA) ratio was used as a biomarker of Zn status. Two groups of chickens (<i>n</i> = 15) were fed different diets: a high-Zn (46.5 μg Zn g^–1) and a low-Zn wheat-based diet (32.8 μg Zn g^–1). Dietary Zn intakes, body weight, serum Zn, and the erythrocyte fatty acid profile were measured, and tissues were taken for gene expression analysis. Serum Zn concentrations were greater in the high Zn group (<i>p</i> < 0.05). Duodenal mRNA expression of various Zn transporters demonstrated expression upregulation in the birds fed a low Zn diet (<i>n</i> = 15, <i>p</i> < 0.05). The LA:DGLA ratio was higher in the birds fed the low Zn diet (<i>p</i> < 0.05). The higher amount of Zn in the biofortified wheat resulted in a greater Zn uptake

Alterations in the Gut (<i>Gallus gallus</i>) Microbiota Following the Consumption of Zinc Biofortified Wheat (<i>Triticum aestivum</i>)‑Based Diet

The structure and function of cecal microbiota following the consumption of a zinc (Zn) biofortified wheat diet was evaluated in a well-studied animal model of human nutrition (<i>Gallus gallus</i>) during a six-week efficacy trial. Using 16S rRNA gene sequencing, a significant increase in β- but not α-microbial diversity was observed in the animals receiving the Zn biofortified wheat diet, relative to the control. No significant taxonomic differences were found between the two groups. Linear discriminant analysis revealed a group of metagenomic biomarkers that delineated the Zn replete versus Zn deficient phenotypes, such that enrichment of lactic acid bacteria and concomitant increases in Zn-dependent bacterial metabolic pathways were observed in the Zn biofortified group, and expansion of mucin-degraders and specific bacterial groups able to participate in maintaining host Zn homeostasis were observed in the control group. Additionally, the <i>Ruminococcus</i> genus appeared to be a key player in delineating the Zn replete microbiota from the control group, as it strongly predicts host Zn adequacy. Our data demonstrate that the gut microbiome associated with Zn biofortified wheat ingestion is unique and may influence host Zn status. Microbiota analysis in biofortification trials represents a crucial area for study as Zn biofortified diets are increasingly delivered on a population-wide scale

DataSheet_1_An accurate, reliable, and universal qPCR method to identify homozygous single insert T-DNA with the example of transgenic rice.zip

Early determination of transgenic plants that are homozygous for a single locus T-DNA insert is highly desirable in most fundamental and applied transgenic research. This study aimed to build on an accurate, rapid, and reliable quantitative real-time PCR (qPCR) method to fast-track the development of multiple homozygous transgenic rice lines in the T1 generation, with low copy number to single T-DNA insert for further analyses. Here, a well-established qPCR protocol, based on the OsSBE4 reference gene and the nos terminator, was optimized in the transgenic Japonica rice cultivar Nipponbare, to distinguish homozygous single-insert plants with 100% accuracy. This method was successfully adapted to transgenic Indica rice plants carrying three different T-DNAs, without any modifications to quickly develop homozygous rice plants in the T1 generation. The accuracy of this qPCR method when applied to transgenic Indica rice approached 100% in 12 putative transgenic lines. Moreover, this protocol also successfully detected homozygous single-locus T-DNA transgenic rice plants with two-transgene T-DNAs, a feature likely to become more popular in future transgenic research. The assay was developed utilizing universal primers targeting common sequence elements of gene cassettes (the nos terminator). This assay could therefore be applied to other transgenic plants carrying the nos terminator. All procedures described here use standardized qPCR reaction conditions and relatively inexpensive dyes, such as SYBR Green, thus the qPCR method could be cost-effective and suitable for lower budget laboratories that are involved in rice transgenic research.</p