Agrobacterium-Mediated Capsicum annuum Gene Editing in Two Cultivars, Hot Pepper CM334 and Bell Pepper Dempsey
| Autor: | Hyun-Ji Jeon, Hyun-Bin Kim, Sung-il Park, Hyeran Kim |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
0301 basic medicine Agrobacterium CaMLO2 Biology Agrobacterium tumefaciens 01 natural sciences Article Catalysis Inorganic Chemistry lcsh:Chemistry Capsicum annuum Dempsey 03 medical and health sciences Pepper Cultivar Physical and Theoretical Chemistry Indel Molecular Biology CRISPR/Cas9 lcsh:QH301-705.5 Spectroscopy Organic Chemistry General Medicine biology.organism_classification Computer Science Applications Horticulture Transformation (genetics) 030104 developmental biology Capsicum annuum CM334 lcsh:Biology (General) lcsh:QD1-999 Callus pBAtC binary vector Solanaceae 010606 plant biology & botany |
| Zdroj: | International Journal of Molecular Sciences, Vol 22, Iss 3921, p 3921 (2021) International Journal of Molecular Sciences Volume 22 Issue 8 |
| ISSN: | 1661-6596 1422-0067 |
| Popis: | Peppers (Capsicum annuum L.) are the most widespread and cultivated species of Solanaceae in subtropical and temperate countries. These vegetables are economically attractive worldwide. Although whole-genome sequences of peppers and genome-editing tools are currently available, the precision editing of peppers is still in its infancy because of the lack of a stable pepper transformation method. Here, we employed three Agrobacterium tumefaciens strainsAGL1, EHA101, and GV3101to investigate which Agrobacterium strain could be used for pepper transformation. Hot pepper CM334 and bell pepper Dempsey were chosen in this study. Agrobacterium tumefaciens GV3101 induced the highest number of calli in cv. Dempsey. All three strains generated similar numbers of calli for cv. CM334. We optimized a suitable concentration of phosphinothricin (PPT) to select a CRISPR/Cas9 binary vector (pBAtC) for both pepper types. Finally, we screened transformed calli for PPT resistance (1 and 5 mg/L PPT for cv. CM334 and Dempsey, respectively). These selected calli showed different indel frequencies from the non-transformed calli. However, the primary indel pattern was consistent with a 1-bp deletion at the target locus of the C. annuum MLO gene (CaMLO2). These results demonstrate the different sensitivity between cv. CM334 and Dempsey to A. tumefaciens-mediated callus induction, and a differential selection pressure of PPT via pBAtC binary vector. |
| Databáze: | OpenAIRE |
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