ArticleFrontiers in plant science2023
A highly efficient genetic transformation system for broccoli and subcellular localization.
Article in Frontiers in plant science, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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10 citing papers in PubMed, 10 citations in OpenAlex.
- Molecular regulators of regeneration and strategies for overcoming genotype-dependent recalcitrance in wheat (Triticum aestivum).Communications biology · 2026Review
- Two Optimized Methods for Efficient, Stable and Transient Transformation of Broccoli (Plants (Basel, Switzerland) · 2026Article
- Study on Genetic Transformation System of Cabbage (Plants (Basel, Switzerland) · 2025Article
- Identification of EXPA4 as a key gene in cotton salt stress adaptation through transcriptomic and coexpression network analysis of root tip protoplasts.BMC plant biology · 2025Article
- Development of an optimized protocol for protoplast-to-plant regeneration of selected varieties of Brassica oleracea L.BMC plant biology · 2024Article
- Improved Protocol for EfficientPlants (Basel, Switzerland) · 2024Article
- Advances in and Perspectives on Transgenic Technology and CRISPR-Cas9 Gene Editing in Broccoli.Genes · 2024Review
- Application and development of CRISPR technology in the secondary metabolic pathway of the active ingredients of phytopharmaceuticals.Frontiers in plant science · 2024Review
- Establishment of a Rapid and EffectivePlants (Basel, Switzerland) · 2023Article
- An Efficient Transformation System for Fast Production ofPlants (Basel, Switzerland) · 2023Article
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Authors and funding
5 authors at 2 institutions in 1 country.
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Abstract
Introduction: Agrobacterium-mediated genetic transformation has been widely used for the identification of functional genes and regulatory and developmental mechanisms in plants. However, there are still some problems of low genetic transformation efficiency and high genotype dependence in cruciferous crops. Methods: In this study, broccoli, a worldwide Brassica crop, was used to investigate the effects of genotype, explant type, concentration of hygromycin B used during seedling selection, overexpression vector type, RNAi and CRISPR/cas9 on the genetic transformation efficiency. At the same time, two vectors, PHG-031350 and PHG-CRa, were used for subcellular localization of the glucoraphanin synthesis-related gene FMOGS-OX5 and clubroot resistance gene by a PEG-Ca2+-mediated transient transformation system for broccoli protoplasts. Finally, the Agrobacterium-mediated genetic transformation system of broccoli was optimized and improved. Results and Discussion: This study showed that hypocotyl explants are more suitable for Agrobacterium-mediated transgene and CRISPR/Cas9 gene editing of broccoli. In contrast to previous studies, we found that 5 mg/L hygromycin B was more advantageous for the selection of resistant broccoli sprouts, and genotype 19B42 reached the highest transformation rate of 26.96%, which is higher than that in Brassica oleracea crops. In addition, the inbred line 19B42 successfully achieved high genetic transformation of overexpression, RNAi and CRISPR/Cas9 vectors; thus, it is powerful recipient material for the genetic transformation of broccoli. Subcellular localization proved that the glucoraphanin metabolism-related gene Bol031350 and clubroot resistance gene CRa were both expressed in the cytoplasm and nucleus, which provided a scientific basis for studying the regulation of glucosinolate metabolism and clubroot resistance in cruciferous crops. Therefore, these findings will provide new insight into the improvement of the genetic transformation and molecular breeding of Brassica oleracea crops.
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