Abstract
Quite a few studies have been conducted to improve the Agrobacterium-mediated transformation of pineapple, which is the second most important commercial tropical fruit crop worldwide. However, pineapple transformation remains challenging, due to technical difficulties, the lengthy regeneration process, and a high labor requirement. There have not been any studies specifically addressing the introduction of GFP-tagged genes into pineapples through Agrobacterium-mediated transformation, which would enable easy, non-destructive expression detection. It would also allow expression localization at the organelle level, which is not possible with GUS a reporter gene that encodes β-glucuronidase or a herbicide resistance reporter gene. Here, we report a method for the introduction of GFP-tagged genes into pineapples through Agrobacterium-mediated transformation. We used embryonic calli for transformation, and plants were regenerated through somatic embryogenesis. In this study, we optimized the incubation time for Agrobacterium infection, the co-cultivation time, the hygromycin concentration for selection, and the callus growth conditions after selection. Our strategy reduced the time required to obtain transgenic plants from 7.6 months to 6.1 months. The expression of GFP-tagged AcWRKY28 was observed in the nuclei of transgenic pineapple root cells. This method allows easy, non-destructive expression detection of transgenic constructs at the organelle level. These findings on pineapple transformation will help accelerate pineapple molecular breeding efforts to introduce new desirable traits.
Highlights
Pineapple is an economically important tropical fruit crop, and it is used as a fiber crop and source of the valuable pharmaceutical enzyme bromelain [1,2]
Among the 54 AcWRKY genes, we focused on AcWRKY28 (Aco005719.1) for the analysis and transformation
The AcWRKY28 open reading frame (ORF) length is 609 bp, and further analysis revealed that the gene is located on chromosome 11 (Figure 1B)
Summary
Pineapple is an economically important tropical fruit crop, and it is used as a fiber crop and source of the valuable pharmaceutical enzyme bromelain [1,2]. Agrobacterium-mediated transformation is challenging in pineapple, and studies aimed at introducing new desired traits to improve pineapple quality have been limited. Some important traits have been introduced into pineapples through. Efforts have been made to improve the transformation process by reducing the time required for plant regeneration via direct regeneration of adventitious buds from infected stem disks and leaf bases [7]. Despite these advances, pineapple transformation is not as fully developed as transformation in other crops, such as rice, due in part to difficulties in the time-consuming regeneration process. Transformation efficiency has been shown to be as low as 0.12–2.26% [7]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
