Agrobacterium tumefaciens-Mediated Plant Transformation: A Review

Molecular Biotechnology, Journal Year: 2023, Volume and Issue: 66(7), P. 1563 - 1580

Published: June 20, 2023

Language: Английский

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Advances in the molecular mechanism of grapevine resistance to fungal diseases

Molecular Horticulture, Journal Year: 2025, Volume and Issue: 5(1)

Published: Jan. 2, 2025

Abstract Grapevine is an important economic fruit tree worldwide, but grape production has been plagued by a vast number of fungal diseases, which affect vigor and the quality yield berries. To seek remedies for such issues, researchers have always committed to conventional biotechnological breeding. In recent years, increasing progress made in elucidating molecular mechanisms grape–pathogenic fungi interactions resistance regulation. Here, we summarize current knowledge on basis grapevine including effector-mediated susceptibility resistance, resistant regulatory networks grapevine, innovative approaches genetic transformation, strategies improve resistance. Understanding exploring accurately regulating diseases.

Language: Английский

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Regeneration of non-chimeric plants from DNA-free edited grapevine protoplasts

Frontiers in Plant Science, Journal Year: 2022, Volume and Issue: 13

Published: Dec. 1, 2022

The application of New Breeding Techniques (NBTs) in

Language: Английский

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Plant protoplasts in the age of synthetic biology

Journal of Experimental Botany, Journal Year: 2023, Volume and Issue: 74(13), P. 3821 - 3832

Published: May 23, 2023

Abstract Protoplasts, which are plant cells with their cell walls removed, have been used for decades in research and instrumental genetic transformation the study of various aspects physiology genetics. With advent synthetic biology, these individualized fundamental to accelerate ‘design–build–test–learn’ cycle, is relatively slow research. Despite potential, challenges remain expanding use protoplasts biology. The capacity individual hybridize form new varieties, regenerate from single cells, creating individuals features underexplored. main objective this review discuss biology highlight exploiting protoplast technologies ‘age biology’.

Language: Английский

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Simultaneous editing of two DMR6 genes in grapevine results in reduced susceptibility to downy mildew

Frontiers in Plant Science, Journal Year: 2023, Volume and Issue: 14

Published: Aug. 21, 2023

The reduction of pesticide treatments is paramount importance for the sustainability viticulture, and it can be achieved through a combination strategies, including cultivation vines ( Vitis vinifera ) that are resistant or tolerant to diseases such as downy mildew (DM). In many crops, knock-out Downy Mildew Resistant 6 DMR6 proved successful in controlling DM-resistance, but effect mutations genes not yet known grapevine. Today, gene editing serves crop improvement with small specific while maintaining genetic background commercially important clones. Moreover, recent technological advances allowed produce non-transgenic grapevine clones by regeneration protoplasts edited CRISPR/Cas9 ribonucleoprotein. This approach may revolutionize production new varieties clones, requires knowledge about targets impact on plant phenotype fitness different cultivars. this work we generated single double mutants susceptibility (S) using CRISPR/Cas9, showed only combined VviDMR6-1 VviDMR6-2 effective reducing DM two table-grape cultivars increasing levels endogenous salicylic acid. Therefore, both necessary control real-world agricultural settings, which could potentially lead unwanted phenotypes. Additional research, trials conducted experimental vineyards, required gain deeper understanding -based resistance.

Language: Английский

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Grapevine gray mold disease: infection, defense and management

Horticulture Research, Journal Year: 2024, Volume and Issue: 11(9)

Published: July 10, 2024

Abstract Grapevine (Vitis vinifera L.,) is among the world’s leading fruit crops. The production of grapes severely affected by many diseases including gray mold, caused necrotrophic fungus Botrytis cinerea. Although all Vitis species can be hosts for B. cinerea, V. are particularly susceptible. Accordingly, this disease poses a significant threat to grape industry and causes substantial economic losses. Development resistant cultivars has progressed from incidental selection farmers, targeted through use statistics experimental design, employment genetic genomic data. Emerging technologies such as marker-assisted engineering have facilitated development that possess resistance A promising method involves using CRISPR/Cas9 system induce mutagenesis develop genetically modified non-transgenic Hence, scientists now engaged in active pursuit identifying genes associated with susceptibility resistance. This review focuses on known mechanisms interaction between cinerea pathogen its grapevine host. It also explores innate immune systems evolved vinifera, objective facilitating rapid cultivars.

Language: Английский

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New improvements in grapevine genome editing: high efficiency biallelic homozygous knock-out from regenerated plantlets by using an optimized zCas9i

Plant Methods, Journal Year: 2024, Volume and Issue: 20(1)

Published: March 18, 2024

Abstract Background For ten years, CRISPR/cas9 system has become a very useful tool for obtaining site-specific mutations on targeted genes in many plant organisms. This technology opens up wide range of possibilities improved breeding the future. In plants, CRISPR/Cas9 is mostly used through stable transformation with constructs that allow expression Cas9 gene and sgRNA. Numerous studies have shown mutation efficiency can vary greatly between different species due to factors such as efficiency, expression, nucleotide sequence, addition intronic sequences, other parameters. Since 2016, when first edited grapevine was created, number using functional genomic approaches remained low difficulties editing efficiency. this study, we optimized process obtain generate knock-out mutants ( Vitis vinifera cv. ‘Chardonnay’). Building existing methods transformation, method selecting transformed plants at chosen steps developing fluorescence microscopy. Results By comparison two promoters, increased maize-codon containing 13 introns (zCas9i), achieving 100% biallelic plantlets ‘Chardonnay’. These results are directly correlated level. Conclusions Taken together, our highlight complete methodology homozygous future programs grapevine.

Language: Английский

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Genome editing of a recalcitrant wine grape genotype by lipofectamine‐mediated delivery of CRISPR/Cas9 ribonucleoproteins to protoplasts

The Plant Journal, Journal Year: 2024, Volume and Issue: 119(1), P. 404 - 412

Published: April 22, 2024

SUMMARY The main bottleneck in the application of biotechnological breeding methods to woody species is due vitro regeneration recalcitrance shown by several genotypes. On other side, species, especially grapevine ( Vitis vinifera L.), use most pesticides and expensive inputs agriculture, making development efficient approaches genetic improvement absolutely urgent. Genome editing an extremely promising technique particularly for wine grape genotypes, as it allows modify desired gene a single step, preserving all quality traits selected appreciated elite varieties. A genome protocol production transgene‐free plants, exploiting lipofectamine‐mediated direct delivery CRISPR–Cas9 ribonucleoproteins (RNPs) target phytoene desaturase gene, reported. We focused on Nebbiolo V. ), recalcitrant genotype used produce outstanding wines, such Barolo Barbaresco. PEG‐mediated method available literature employed highly embryogenic genotypes did not allow proper embryo Nebbiolo. Lipofectamines, contrary, have negative impact protoplast viability plant regeneration, leading obtainment fully developed edited plants after about 5 months from transfection. Our work represents one first examples lipofectamine delivering reagents protoplasts. important result achieved could be extended varieties species.

Language: Английский

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The Role of Italy in the Use of Advanced Plant Genomic Techniques on Fruit Trees: State of the Art and Future Perspectives

International Journal of Molecular Sciences, Journal Year: 2023, Volume and Issue: 24(2), P. 977 - 977

Published: Jan. 4, 2023

Climate change is deeply impacting the food chain production, lowering quality and yield. In this context, international scientific community has dedicated many efforts to enhancing resilience sustainability in agriculture. Italy among main European producers of several fruit trees; therefore, national research centers universities undertook initiatives maintain specificity ‘Made Italy’ label. Despite their importance, crops are suffering from difficulties associated with conventional breeding approaches, especially terms financial commitment, land resources availability, long generation times. The ‘new genomic techniques’ (NGTs), renamed as ‘technologies for assisted evolution’ (TEAs), reduce time required obtain genetically improved cultivars while precisely targeting specific DNA sequences. This review aims illustrate role Italian use NGTs, a focus on Citrus, grapevine, apple, pear, chestnut, strawberry, peach, kiwifruit. For each crop, key genes traits which working, well technological improvements advancements regeneration local varieties, presented. Lastly, placed legal aspects contexts.

Language: Английский

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Editing VvDXS1 for the creation of muscat flavour in Vitis vinifera cv. Scarlet Royal

Plant Biotechnology Journal, Journal Year: 2024, Volume and Issue: 22(6), P. 1610 - 1621

Published: Jan. 20, 2024

Muscat flavour represents a group of unique aromatic attributes in some grape varieties. Biochemically, berries with muscat produce high levels monoterpenes. Monoterpene biosynthesis is mainly through the DOXP/MEP pathway, and VvDXS1 encodes first enzyme this plastidial pathway terpene grapevine. A single-point mutation resulting substitution lysine an asparagine at position 284 protein has previously been identified as major cause for producing grapes. In study, same was successfully created prime editing table Vitis vinifera cv. 'Scarlet Royal'. The targeted point detected most transgenic vines, varying efficiencies. No unintended mutations were edited alleles, either by PCR Sanger sequencing or amplicon sequencing. More than dozen vines efficiency more 50%, indicating that these likely derived from single cells which one allele edited. These had much higher monoterpenes their leaves control, similar to what found leaf samples between field-grown non-muscat

Language: Английский

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