How CRISPR could yield the next blockbuster crop

Nature, Год журнала: 2024, Номер 625(7994), С. 230 - 232

Опубликована: Янв. 9, 2024

Язык: Английский

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Site‐directed genotype screening for elimination of antinutritional saponins in quinoa seeds identifies TSARL1 as a master controller of saponin biosynthesis selectively in seeds

Plant Biotechnology Journal, Год журнала: 2024, Номер 22(8), С. 2216 - 2234

Опубликована: Апрель 4, 2024

Summary Climate change may result in a drier climate and increased salinization, threatening agricultural productivity worldwide. Quinoa ( Chenopodium quinoa ) produces highly nutritious seeds tolerates abiotic stresses such as drought high salinity, making it promising future food source. However, the presence of antinutritional saponins their is an undesirable trait. We mapped genes controlling seed saponin content to genomic region that includes TSARL1 . isolated desired genetic variation this gene by producing large mutant library commercial cultivar screening for specific nucleotide substitutions using droplet digital PCR. were able rapidly isolate two independent tsarl1 mutants, which retained leaves roots defence, but undetectable coat. further could show specifically controls biosynthesis committed step after 2,3‐oxidosqualene. Our work provides new important knowledge on function represents breakthrough breeding.

Язык: Английский

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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

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Harnessing the Power of an Extensive EMS‐Induced Sorghum Population for Rapid Crop Improvement

Physiologia Plantarum, Год журнала: 2024, Номер 176(4)

Опубликована: Июль 1, 2024

Plant breeders leverage mutagenesis using chemical, biological, and physical mutagens to create novel trait variations. Many widely used sorghum genotypes have a narrow genetic base, which hinders improvements classical breeding. Enhancing the diversity of genome thus remains key priority for breeders. To accelerate enhancement sorghum, an extensive library comprised seeds from 150,000 individual mutant plants Sorghum bicolor inbred line BTx623 was established ethyl methanesulphonate (EMS) as mutagen. The bulked into 1498 pools (~100 seed heads per pool). In each pool, DNA extracted subset screened FIND-IT technology based on droplet digital PCR. All 43 nucleotide substitutions that were identified, demonstrating potential identify any EMS-derived mutation in elite within days. This diverse represents largest collection mutants ever conceived, estimated cover 240% all possible EMS-induced points genome. Using FIND-IT, speed at specific desired can be identified is major upgrade conventional reverse techniques. Additionally, ease valuable variants integrated commercial lines far simpler less expensive process compared editing. Genomic variations will direct utility breeding resource applications, allowing enhanced adaptation climate change yield marginal environments.

Язык: Английский

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Isolation and characterization of haploid heterothallic beer yeasts

Applied Microbiology and Biotechnology, Год журнала: 2025, Номер 109(1)

Опубликована: Янв. 22, 2025

Abstract Improving ale or lager yeasts by conventional breeding is a non-trivial task. Domestication of yeasts, which are hybrids between Saccharomyces cerevisiae and eubayanus , has led to evolved strains with severely reduced abolished sexual reproduction capabilities, due to, e.g. postzygotic barriers. On the other hand, S. particularly Kveik yeast strains, were shown produce abundant viable spores (~ 60%; Dippel et al. Microorganisms 10(10):1922, 2022). This us investigate usefulness for breeding. Surprisingly, we could isolate heterothallic colonies from germinated different strains. These presented stable mating types in confrontation assays pheromone-sensitive tester Heterothallism was inactivating mutations their HO genes. amino acid exchanges Ho protein, revealing known G223D mutation also novel G217R mutation, both type switching. We generated MAT MATα lines four named Odin, Thor, Freya Vör. Analyses bud scar positions these revealed axial bipolar budding patterns. However, ability Vör form meiotic offspring haploid demonstrated that haploid. Fermentation analyses indicated all able ferment maltose maltotriose. Odin found share not only gene, but PAD1 FDC1 genes makes this strain POF-, i.e. generate phenolic off-flavours, key feature yeasts. yeast-derived may open avenues generating selection utilizing power genetics, thus lifting block domestication brought about. Key points • Haploid isolated. Heterothallic bear mutant alleles leading change. One be POF- gene rendering it negative off-flavor production. highly accessible beer improvements breeding, employing genetics regimes.

Язык: Английский

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Starch biosynthesis and crop bioengineering

Grain & Oil Science and Technology, Год журнала: 2025, Номер unknown

Опубликована: Апрель 1, 2025

Язык: Английский

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Sodium azide mutagenesis induces a unique pattern of mutations

PLoS Genetics, Год журнала: 2025, Номер 21(6), С. e1011634 - e1011634

Опубликована: Июнь 3, 2025

The nature and effect of mutations are fundamental importance to the evolutionary process. generation with mutagens has also played important roles in genetics. Applications include dissecting genetic basis trait variation, inducing desirable traits crops, understanding load. Previous studies sodium azide-induced have reported single nucleotide variants (SNVs) found individual genes. To characterize induced by azide, we analyze whole-genome sequencing (WGS) 11 barley lines derived from azide mutagenesis, where all were selected for diminution plant fitness owing mutations. We contrast observed mutagen-induced those standing variation WGS 13 landraces. Here, report indels that two orders magnitude more abundant than expected based on nominal mutation rates. SNVs very specific, C → T changes occurring a context followed another same strand (or reverse complement). codons most affected mutagen azide-specific CC motif complement), resulting handful amino acid few stop codons. specific suggests could be chosen experimental goals. Sodium would not ideal gene knockouts but will create many missense subtle effects protein function.

Язык: Английский

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Is it the end of TILLING era in plant science?

Frontiers in Plant Science, Год журнала: 2023, Номер 14

Опубликована: Авг. 22, 2023

Since its introduction in 2000, the TILLING strategy has been widely used plant research to create novel genetic diversity. is based on chemical or physical mutagenesis followed by rapid identification of mutations within genes interest. mutants may be for functional analysis and being nontransgenic, they directly pre-breeding programs. Nevertheless, classical a random process, giving rise all over genome. Therefore carry background mutations, some which affect phenotype should eliminated, often time-consuming. Recently, new strategies targeted genome editing, including CRISPR/Cas9-based methods, have developed optimized many species. These methods precisely target only interest produce very few off-targets. Thus, question arises: it end era studies? In this review, we recap basics strategy, summarize current status present recent achievements. Based these reports, conclude that still plays an important role as valuable tool generating variation genomics breeding projects.

Язык: Английский

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HvSL1andHvMADS16promote stamen identity to restrict multiple ovary formation in barley

Journal of Experimental Botany, Год журнала: 2023, Номер 74(17), С. 5039 - 5056

Опубликована: Июнь 3, 2023

Correct floral development is the result of a sophisticated balance molecular cues. Floral mutants provide insight into main genetic determinants that integrate these cues, as well providing opportunities to assess functional variation across species. In this study, we characterize barley (Hordeum vulgare) multiovary mov2.g and mov1, propose causative gene sequences: C2H2 zinc-finger HvSL1 B-class HvMADS16, respectively. absence HvSL1, florets lack stamens but exhibit supernumerary carpels, resulting in multiple grains per floret. Deletion HvMADS16 mov1 causes homeotic conversion lodicules bract-like organs carpels contain non-functional ovules. Based on developmental, genetic, data, model by which stamen specification defined acting upstream HvMADS16. The present work identifies strong conservation formation pathways with other cereals, also reveals intriguing species-specific differences. findings lay foundation for better understanding architecture Triticeae, key target crop improvement.

Язык: Английский

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Optimized barley phytase gene expression by focused FIND-IT screening for mutations in cis-acting regulatory elements

Frontiers in Plant Science, Год журнала: 2024, Номер 15

Опубликована: Март 1, 2024

Introduction Induced modification of plant gene expression is both fundamental and applied importance. Cis-acting regulatory elements (CREs) are major determinants the spatiotemporal strength expression. Yet, there few examples where induced genetic variation in predetermined CREs has been exploited to improve or investigate crop plants. Methods The digital PCR based FIND-IT technology was discover barley mutants with CRE variants promoter nutritional important grain phytase ( PAPhy_a ) gene. Results discussion Mutants higher lower ultimately mature activity (MGPA), respectively, were discovered. Field trials inositol phosphate profiling during germination showed that does not influence agronomic performance under trial conditions but it shorten lag time mobilization germination. Higher endogenous MGPA an improvement quality for feed use as improves bioavailability monogastric animals. Moreover, targeted motifs shared a range seed expressed genes like key cereal legume storage genes, current results demonstrates concept modulating individual levels genes.

Язык: Английский

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