A CRISPR/Cas9-Based Mutagenesis Protocol for Brachypodium distachyon and Its Allopolyploid Relative, Brachypodium hybridum

Hus, Karolina; Betekhtin, Alexander; Piński, Artur; Rojek-Jelonek, Magdalena; Grzebelus, Ewa; Nibau, Cândida; Gao, Ming‐Jun; Jaeger, Katja E.; Jenkins, Glyn; Doonan, John; Hasterok, Robert

doi:10.3389/fpls.2020.00614

Cited by 14 publications

(11 citation statements)

References 51 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, this requires detailed knowledge about the molecular mechanisms that underlie the various stress responses [8][9][10][11][12]. In order to facilitate the development of climate-smart crops, Brachypodium distachyon was proposed as a model plant for temperate grasses [13,14] because it offers a suite of advantages: undemanding growth requirements, small stature, short life cycle, relatively small genome and established protocols for efficient genomic editing [3,[15][16][17][18]. Large collections of natural accessions with well-described genetic diversity and variation in DNA methylation are also available [19,20].…”

Section: Introductionmentioning

confidence: 99%

Changes in the Cell Wall Proteome of Leaves in Response to High Temperature Stress in Brachypodium distachyon

Piński

Betekhtin

Skupień-Rabian

et al. 2021

IJMS

Self Cite

View full text Add to dashboard Cite

High temperature stress leads to complex changes to plant functionality, which affects, i.a., the cell wall structure and the cell wall protein composition. In this study, the qualitative and quantitative changes in the cell wall proteome of Brachypodium distachyon leaves in response to high (40 °C) temperature stress were characterised. Using a proteomic analysis, 1533 non-redundant proteins were identified from which 338 cell wall proteins were distinguished. At a high temperature, we identified 46 differentially abundant proteins, and of these, 4 were over-accumulated and 42 were under-accumulated. The most significant changes were observed in the proteins acting on the cell wall polysaccharides, specifically, 2 over- and 12 under-accumulated proteins. Based on the qualitative analysis, one cell wall protein was identified that was uniquely present at 40 °C but was absent in the control and 24 proteins that were present in the control but were absent at 40 °C. Overall, the changes in the cell wall proteome at 40 °C suggest a lower protease activity, lignification and an expansion of the cell wall. These results offer a new insight into the changes in the cell wall proteome in response to high temperature.

show abstract

Section: Introductionmentioning

confidence: 99%

Changes in the Cell Wall Proteome of Leaves in Response to High Temperature Stress in Brachypodium distachyon

Piński

Betekhtin

Skupień-Rabian

et al. 2021

IJMS

Self Cite

View full text Add to dashboard Cite

show abstract

“…Developing efficient protocol for buckwheat will help to generate plant varieties with improved traits such as better resistance to biotic and abiotic stresses and improved breeding timelines. CRISPR/Cas9 gene editing technology was successfully used in Brachypodium [131], Arabidopsis thaliana, sorghum [132], rice [133], barley [134], wheat [135] and corn [136], but not in Fagopyrum species. The CRISPR/Cas9 system can also be used in targeting microRNAs (miRNA) in order to gain insight into miRNA regulatory pathways.…”

Section: Future Perspectivesmentioning

confidence: 99%

Buckwheat in Tissue Culture Research: Current Status and Future Perspectives

Tomasiak

Zhou

Betekhtin

2022

IJMS

Self Cite

View full text Add to dashboard Cite

Buckwheat is a member of a genus of 23 species, where the two most common species are Fagopyrum esculentum (common buckwheat) and Fagopyrum tataricum (Tartary buckwheat). This pseudocereal is a source of micro and macro nutrients, such as gluten-free proteins and amino acids, fatty acids, bioactive compounds, dietary fibre, fagopyrins, vitamins and minerals. It is gaining increasing attention due to its health-promoting properties. Buckwheat is widely susceptible to in vitro conditions which are used to study plantlet regeneration, callus induction, organogenesis, somatic embryogenesis, and the synthesis of phenolic compounds. This review summarises the development of buckwheat in in vitro culture and describes protocols for the regeneration of plantlets from various explants and differing concentrations of plant growth regulators. It also describes callus induction protocols as well as the role of calli in plantlet regeneration. Protocols for establishing hairy root cultures with the use of Agrobacterium rhizogens are useful in the synthesis of secondary metabolites, as well as protocols used for transgenic plants. The review also focuses on the future prospects of buckwheat in tissue culture and the challenges researchers are addressing.

show abstract

“…Inactivation or knock-out of the PDS gene disrupts chlorophyll and carotenoid biosynthesis and result in albino and dwarf plants ( Tian, 2015;Kaur et al, 2018). Therefore, the PDS gene has been used extensively as a molecular and morphological marker for the demonstration of genome editing in several plant species (Hsu et al, 2019;Wilson et al, 2019;Hus et al, 2020) including tomato (Pan et al, 2016;Parkhi et al, 2018).…”

Section: Crispr/cas9 Vector Construction and Tomato Transformation Wimentioning

confidence: 99%

“…Li et al, 2018b;X. Li et al, 2018;Tashkandi et al, 2018;Tomlinson et al, 2019;Yin et al, 2018;Zhang et al, 2018;Hus et al, 2020). However, almost all these studies used model tomato cultivars such as Micro-Tom and M-82 (Brooks et al, 2014;Čermák et al, 2015) with limited use for breeders and farmers.…”

Section: Crispr/cas9 Vector Construction and Tomato Transformation Wimentioning

confidence: 99%

Optimization of Agrobacterium-mediated transformation and regeneration for CRISPR/ Cas9 genome editing of commercial tomato cultivars

Seçgin¹,

Kavas²,

Yıldırım³

2021

Turk J Agric For

View full text Add to dashboard Cite

Tomato (Solanum lycopersicum) is the second most important horticultural crop worldwide that is widely used as a model plant in genetic manipulation of Solanaceae. CRISPR/Cas9 system has been successfully utilized in several studies for genome edition of model tomato cultivars. However, these genome editing systems should be also optimized for commercial tomato cultivar for direct application of genome editing in field conditions. In this study, we have optimized an Agrobacterium-mediated gene transfer and regeneration system for CRISPR/Cas9 genome editing in two commercial tomato cultivars for the first time. The effect of explant type, genotype, pre-transformation time, Agrobacterium concentration, infection time, and different co-culture periods of bacteria were evaluated to optimize the regeneration and transformation parameters. The highest regeneration capacity of 83 % was obtained from cotyledons of Crocker incubated in a medium supplemented with BA (3 mg/L) and IAA (0.1 mg/L). The maximum transformation frequency was obtained by using the following parameters: cotyledon explants of commercial Crocker cultivar that were left for 2 days of pre-transformation incubation, infected with Agrobacterium for 10 minutes at a concentration of OD 600 of 0.6 and co-cultivated with Agrobacterium cells for 48 hours. CRISPR/Cas9 system was tested with two gRNAs targeting the phytoene desaturase gene.Fully albino and chimeric plants were successfully produced with optimized transformation and culture conditions in up to 71% of all regenerated plants. In the current study, we optimized the implementation of the CRISPR/Cas9 technique in a commercial tomato cultivar and our method will enable breeders to make necessary changes in traits of interest to improve tomato crop for commercial applications.

show abstract

A CRISPR/Cas9-Based Mutagenesis Protocol for Brachypodium distachyon and Its Allopolyploid Relative, Brachypodium hybridum

Cited by 14 publications

References 51 publications

Changes in the Cell Wall Proteome of Leaves in Response to High Temperature Stress in Brachypodium distachyon

Changes in the Cell Wall Proteome of Leaves in Response to High Temperature Stress in Brachypodium distachyon

Buckwheat in Tissue Culture Research: Current Status and Future Perspectives

Optimization of Agrobacterium-mediated transformation and regeneration for CRISPR/ Cas9 genome editing of commercial tomato cultivars

Contact Info

Product

Resources

About