Accelerating Cereal Breeding for Disease Resistance Through Genome Editing

Reddy, M. Raghurami; Acaso, Joan T.; Alakonya, Amos; Mangrauthia, Satendra K.; Sundaram, R. M.; Balachandran, S. M.; Biswal, Akshaya Kumar

doi:10.1007/978-981-19-0600-8_15

Cited by 1 publication

(1 citation statement)

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“…Gene editing directly in commercial cultivars is a viable alternative to expedite trait improvement, particularly for food security ( Dhugga, 2022 ; Pixley et al., 2022 ). A recently introduced technology, Clustered Regularly Interspaced Short Palindromic Repeats and its associated endonucleases (CRISPR/Cas), has revolutionized the field of gene editing, which was slow to perform with the previously available technologies ( Raghurami-Reddy et al., 2022 ). However, wheat lags other cereals such as rice and maize for its genetic manipulation through transformation, probably because of its complex genome.…”

Section: Introductionmentioning

confidence: 99%

An efficient transformation method for genome editing of elite bread wheat cultivars

et al. 2023

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An efficient genetic transformation protocol is necessary to edit genes for trait improvement directly in elite bread wheat cultivars. We used a protein fusion between a wheat growth-regulating factor 4 (GRF4) and its interacting factor (GIF1) to develop a reproducible genetic transformation and regeneration protocol, which we then used to successfully transform elite bread wheat cultivars Baj, Kachu, Morocco, Reedling, RL6077, and Sujata in addition to the experimental cultivar Fielder. Immature embryos were transformed with the vector using particle bombardment method. Transformation frequency increased nearly 60-fold with the GRF4-GIF1-containing vectors as compared to the control vector and ranged from ~5% in the cultivar Kachu to 13% in the cultivar RL6077. We then edited two genes that confer resistance against leaf rust and powdery mildew directly in the aforementioned elite cultivars. A wheat promoter, TaU3 or TaU6, to drive the expression of guide RNA was effective in gene editing whereas the OsU3 promoter failed to generate any edits. Editing efficiency was nearly perfect with the wheat promoters. Our protocol has made it possible to edit genes directly in elite wheat cultivars and would be useful for gene editing in other wheat varieties, which have been recalcitrant to transformation thus far.

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Section: Introductionmentioning

confidence: 99%