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Open AccessArticle

An Efficient Agrobacterium Mediated Transformation of Pineapple with GFP-Tagged Protein Allows Easy, Non-Destructive Screening of Transgenic Pineapple Plants

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Key Lab of Genetics, Breeding and Multiple Utilization of Crops, Ministry of Education, Fujian Provincial Key Laboratory of Haixia Applied Plant Systems Biology, State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, Center for Genomics and Biotechnology, College of Agriculture, Fujian Agriculture and Forestry University, Fuzhou 350002, China
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College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China
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College of Forestry, Fujian Agriculture and Forestry University, Fuzhou 350002, China
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State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi Key Lab of Sugarcane Biology, College of Agriculture, Guangxi University, Nanning 530004, Guangxi, China
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Author to whom correspondence should be addressed.
Biomolecules 2019, 9(10), 617; https://doi.org/10.3390/biom9100617
Received: 11 September 2019 / Revised: 11 October 2019 / Accepted: 15 October 2019 / Published: 17 October 2019
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. View Full-Text
Keywords: pineapple; Agrobacterium; transformation; GFP-tagged protein; WRKY genes pineapple; Agrobacterium; transformation; GFP-tagged protein; WRKY genes
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MDPI and ACS Style

Priyadarshani, S.V.G.N.; Cai, H.; Zhou, Q.; Liu, Y.; Cheng, Y.; Xiong, J.; Patson, D.L.; Cao, S.; Zhao, H.; Qin, Y. An Efficient Agrobacterium Mediated Transformation of Pineapple with GFP-Tagged Protein Allows Easy, Non-Destructive Screening of Transgenic Pineapple Plants. Biomolecules 2019, 9, 617.

AMA Style

Priyadarshani SVGN, Cai H, Zhou Q, Liu Y, Cheng Y, Xiong J, Patson DL, Cao S, Zhao H, Qin Y. An Efficient Agrobacterium Mediated Transformation of Pineapple with GFP-Tagged Protein Allows Easy, Non-Destructive Screening of Transgenic Pineapple Plants. Biomolecules. 2019; 9(10):617.

Chicago/Turabian Style

Priyadarshani, S. V.G.N.; Cai, Hanyang; Zhou, Qiao; Liu, Yanhui; Cheng, Yan; Xiong, Junjie; Patson, Dikoko L.; Cao, Shijiang; Zhao, Heming; Qin, Yuan. 2019. "An Efficient Agrobacterium Mediated Transformation of Pineapple with GFP-Tagged Protein Allows Easy, Non-Destructive Screening of Transgenic Pineapple Plants" Biomolecules 9, no. 10: 617.

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