Just FIND‐IT: Harnessing the true power of induced mutagenesis

Plant Biotechnology Journal, Год журнала: 2024, Номер 22(11), С. 3051 - 3053

Опубликована: Июль 9, 2024

In nature, genetic variation occurs in every population and results the evolution of a diversity new properties, some which promote survival species. To accelerate nature's based on diversity, plant breeders may induce additional mutations to raise number variations increasing chances obtain varieties with desired traits like improved nutritive quality, yields resilience biotic abiotic stress factors. Induced mutagenesis chemical mutagens is considered non-GM has been used barley (Hordeum vulgare) for decades (Hansson et al., 2024). Reverse techniques including TILLING (Targeting Local Lesions Genomes) screening methodology more recently TILLING-by-sequencing spinoffs are tools identify individual variants valuable genomic alterations. However, these hampered by low mutation capacity. PCR-based technique designed detect mismatched single nucleotides target gene. 2023, Szarejko her research group Poland published thorough overview success stories within last 20 years (Szurman-Zubrzycka 2023) description populations different cultivars landraces obtained following (Figure 1a). The sizes range between 1372 9600 variants. frequencies individually chosen dose-dependent (1/154–1/2500 Kbp). When multiplied (# individuals × per individual), total present ranges 10 100 million This sound lot, but genome size around 4300 Mbp (here, RGT Planet; Jayakodi 2020), less than 2% entire mutated. severely reduces possibility find populations. FIND-IT technology approach overriding constraints. was Science Advances 2022 1b) (Knudsen 2022) provides an agile high-throughput screen unprecedented large chemically induced variant combines systematic sample pooling splitting high-sensitivity, droplet digital PCR (ddPCR)–based genotyping targeted identification at single-nucleotide resolution. ddPCR 1000-fold sensitive conventional PCR. applicable any living organism that can be grown field or culture. experimental outlined detail Knudsen illustrated schematically Figure 1b. total, 500 000 plants today available screening. were also developed other crops microorganisms using sodium azide ethyl methanesulphonate (EMS) as mutagens. Thus key sweet seeds white lupin (Mancinotti 2023), eliminate presence anti-nutritional saponins quinoa (Trinh 2024), improve phosphate bioavailability grain (Madsen avoid hydroxynitrile glucoside-derived formation pro-carcinogen carbamate whisky production S1; Jørgensen 2024) modify flavour profiles Saccharomyces species use industrial brewing (Stovisek pipeline knowledge mutant load spectrum doses monitored whole sequencing 1b; 2022). introduced found equally distributed over seven chromosomes higher observed increased mutagen doses. Thus, libraries generated purposes: Medium (e.g. 1.7 mm treatment average 14 770 SNPs plant) gene-function analyses 0.3 5565 breeding whole-genome documents 15% transversions 85% transitions preference C > T G A 1c). Planet genome, 1900 Mb C's G's. Upon dose, randomly positioned each 8000 will transitions. Using technology, library collection 350 now analysed containing approximately = 2 800 1b). Because only harbours nucleotides, this means virtually all putative sites have saturated. saturated, their status shifts from being random become defined distinct one collection. Accordingly, grains carrying specific SNP gene identified many pools TaqMan™ assay You know there just FIND-IT. Compared existing cereal resources relation natural pan-genome accession panel, off-target pressure efficiently reduced backcrossing parent direct crosses elite commercial programmes. context, it noticed would typically adjusted comparable classical step two parents naturally genomes As proof principle, we demonstrated efficiency isolating knockout lines dozen amino acid exchanges miRNA promoter S2; Taking advantage fact non-GMO method, data directly verified growing 2022), important requirement validate crop (Khaipho-Burch 2023). current development high-quality allows strategies highly customized. While static regarding variant, CRISPR technologies still facing multiple challenges field-applicable (Cardi stays agile, offers high flexibility high-throughput, compatible today. regularly updated lines, customized winter versus spring libraries) while sensitivity keeps competitive. isolated line load, original implemented protocols lose during yield cycles providing fast market rollout novel traits. We thank Carlsberg Foundation funding support: grant: CF14-0461 (to B.L.M.) CF15-0236 B.S.). authors not declared conflict interest. Data sharing article no datasets study. S1. Disentangling glucoside biosynthesis metabolon access malting barleys free production. S2. Barley selected agronomically evaluated. Please note: publisher responsible content functionality supporting information supplied authors. Any queries (other missing content) should directed corresponding author article.

Язык: Английский

Impact of kdcA, pdhD, and codY Gene Regulation in Lactococcus lactis 408 on 3-Methylbutanal Formation During Cheddar Cheese Ripening

Food Microbiology, Год журнала: 2025, Номер 130, С. 104768 - 104768

Опубликована: Март 7, 2025

Язык: Английский