Development and validation of PCR marker array for molecular selection towards spring, vernalization-independent and winter, vernalization-responsive ecotypes of white lupin (Lupinus albus L.)

Scientific Reports, Год журнала: 2025, Номер 15(1)

Опубликована: Янв. 21, 2025

Abstract White lupin ( Lupinus albus L.) is an ancient grain legume that still undergoing improvement of domestication traits, including vernalization-responsiveness, providing frost tolerance and preventing winter flowering in autumn-sowing agriculture, vernalization-independence, conferring drought escape by rapid spring-sowing. A recent genome-wide association study highlighted several loci significantly associated with the most contrasting phenotypes, deletions promoter FLOWERING LOCUS T homolog, LalbFTc1 , some DArT-seq/silicoDArT loci. The present aimed to develop validate a versatile PCR marker array enabling molecular selection spring- winter-type white ecotypes. Candidate DArT-seq silicoDArT were transformed into cleaved amplified polymorphic sequence (CAPS) or derived CAPS markers. Developed markers, together those previously published for INDELs quantitative trait from linkage maps, implemented screening germplasm panel subjected 2-year phenotyping phenology traits. Three DArT-seq, two seven INDEL markers positively validated, constituting convenient PCR-based assay accurate reselection towards early thermoneutrality late as well tracking high genetic phenotypic diversity within landraces, revealed study.

Язык: Английский

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Recent Advances in Molecular Tools and Pre-Breeding Activities in White Lupin (Lupinus albus)

Plants, Год журнала: 2025, Номер 14(6), С. 914 - 914

Опубликована: Март 14, 2025

The higher adaptation of landraces to local agroclimatic conditions resulting from natural and moderate artificial selection by farmers within specific environments makes them a crucial source alleles genotypes for cultivation breeding programs. Unlike modern cultivars, which have been developed under more intense selective pressures, exhibit broader genetic base that has documented in landrace collections many crops. This review provides an overview the importance resource valorisation legume species, focusing on cultivated species Lupinus genus, particularly white lupin (Lupinus albus). On one hand, legumes, including Lupins, are considered alternative protein framework sustainable agriculture. other they often neglected terms efforts, despite receiving increasing attention recent years. Here, we also report latest advances development genomic tools, such as novel pangenome identification markers loci target traits, tolerance alkaline soils, can effectively support albus, especially introgression desirable locally adapted varieties.

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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Hazard Identification of Food-Relevant Lupine Alkaloids Focusing on In Vitro Genotoxicity and Mutagenicity

ACS Food Science & Technology, Год журнала: 2025, Номер unknown

Опубликована: Янв. 22, 2025

Recently, it was shown that food products made from the seeds of Lupinus species contain quinolizidine alkaloids in range several mg/kg. Some these compounds are not very well characterized regard to their toxic mechanism action. Specifically, we identified data gaps on genotoxicity and mutagenicity contaminants, which aimed close. Therefore, applied a basic test battery vitro assays including bacterial reverse mutation assay micronucleus study genotoxic potential five selected lupine angustifoline, lupanine, 13-hydroxylupanine, lupinine, sparteine. We found none showed any cytotoxic, genotoxic, or mutagenic vitro.

Язык: Английский

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Alkaloids Derived from Lysine: Quinolizidine Alkaloids, a Focus on Lupin Alkaloids

Springer eBooks, Год журнала: 2025, Номер unknown, С. 1 - 26

Опубликована: Янв. 1, 2025

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Occurrence of quinolizidine alkaloids and phomopsin A in lupin-containing foods: a two-year survey of the Belgian market

Food Additives & Contaminants Part A, Год журнала: 2025, Номер unknown, С. 1 - 16

Опубликована: Апрель 28, 2025

Lupin is a pulse crop valued for its protein content and potential as functional ingredient, e.g. in gluten-free foods. The presence of quinolizidine alkaloids (QAs) phomopsins (PHO) has raised concerns, yet information regarding their occurrence remains limited. A two-year screening QAs PHO lupin-containing foods available on the Belgian market was conducted, based study risk-based sampling plan (n = 339). UHPLC-MS/MS method validated simultaneous quantification seven (i.e. sparteine, lupanine, lupinine, 13α-OH-lupanine, angustifoline, multiflorine albine) phomopsin relevant matrices. not detected any samples (

Язык: Английский

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Beyond co-expression: pathway discovery for plant pharmaceuticals

Current Opinion in Biotechnology, Год журнала: 2024, Номер 88, С. 103147 - 103147

Опубликована: Июнь 3, 2024

Plant natural products have been an important source of medicinal molecules since ancient times. To gain access to the whole diversity these for pharmaceutical applications, it is understand their biosynthetic origins. Whilst co-expression a reliable tool identifying gene candidates, variety complementary methods can aid in screening or refining candidate selection. Here, we review recently employed plant pathway discovery approaches, and highlight future directions field.

Язык: Английский

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The genome of Lupinus mutabilis: Evolution and genetics of an emerging bio‐based crop

The Plant Journal, Год журнала: 2024, Номер unknown

Опубликована: Сен. 12, 2024

SUMMARY Lupinus mutabilis is an under‐domesticated legume species from the Andean region of South America. It belongs to New World lupins clade, which groups several lupin displaying large genetic variation and adaptability highly different environments. L. attracting interest as a potential multipurpose crop diversify European supply plant proteins, increase agricultural biodiversity, fulfill bio‐based applications. This study reports first high‐quality genome assembly, also sequenced assembly species. Through comparative genomics phylogenetics, evolution within legumes described, highlighting both genomic similarities patterns specific , potentially linked environmental adaptations. Furthermore, was used genetics underlying important traits for establishment novel crop, including protein quinolizidine alkaloids contents in seeds, classic resistance genes, properties mycorrhiza‐related genes. These analyses pointed out copy number variation, differential gene contexts, family expansion through tandem duplications likely drivers diversity observed these between other legumes. Overall, will be valuable resource conduct research enable genomic‐based breeding approaches turn into crop.

Язык: Английский

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Metabolic engineering of narrow-leafed lupin for the production of enantiomerically pure (‒)-sparteine

Опубликована: Апрель 26, 2024

Sparteine is a plant-derived alkaloid widely known for its utility as chiral ligand in asymmetric synthesis 1-3 . However, variable market price and availability have failed to meet the demand cheap reliable product 4-6 naturally synthesized by sub-group of legume plants, which typically accumulate complex mixtures closely related alkaloids. Here, we identified two enzymes from narrow-leafed lupin (NLL, L. angustifolius ) that can sequentially oxidize (−)-sparteine (+)-lupanine. The first enzyme cytochrome P450 monooxygenase belonging family 71 (CYP71D189) second one short-chain dehydrogenase/reductase (SDR1). We also screened non-GMO NLL mutant library isolated knockout CYP71D189. displayed an altered metabolic profile where accounted 96% content seeds (GC-MS basis). was enantiomerically pure (99% ee ). Apart profile, did not any noticeable phenotype. Our work demonstrates precursor most QAs provides convenient source this valuable compound academia industry.

Язык: Английский

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Nonsymmetric formation of Δ1-piperideine from lysine in plants via a bacterial-like PLP-dependent enzyme

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Июнь 4, 2024

Abstract Piperidine alkaloids in plants derive from lysine via Δ 1 -piperideine. We investigated the alkaloid biosynthesis Flueggea suffruticosa , a plant that produces diverse piperidine alkaloids, including securinine. Until now, pathways leading to formation of -piperideine have involved two enzymatic steps: decarboxylase and an oxidase, proceeding free cadaverine. discovered synthase (PS), PLP-dependent enzyme forms directly oxidative deamination. demonstrate PS catalyses this transformation non-symmetric manner, show it is positioned within group III family, which typically associated with prokaryotes. also identified paralog gene exhibiting activity. This work delineates first step securinine as unprecedented deamination lysine. It marks identification eukaryotic resolves longstanding question nitrogen incorporation alkaloids.

Язык: Английский

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