Improved protocol for the transformation of adult Citrus sinensis Osbeck ‘Tarocco’ blood orange tissues
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The production of transgenic citrus plants from adult tissues is difficult because of low regeneration and transformation rates. To increase the transformation efficiency of adult citrus tissues, an improved protocol involving adult Citrus sinensis Osbeck ‘Tarocco’ blood orange tissues was developed. Explants were pre-incubated in a liquid medium prior to infection by Agrobacterium tumefaciens. Plant materials were also incubated on callus-induction medium supplemented with various combinations of cytokinin (Cyt) and kanamycin (Kan). An appropriate pre-incubation of the explants increased the transformation efficiency of adult tissues. During the callus-induction period, the Cyt type and Kan concentration had the largest and smallest effects on the transformation efficiency, respectively. The most effective combination of plant growth regulator and Kan for the transformation of ‘Tarocco’ blood orange tissues was 2 mg L−1 2-isopentenyl adenine and 50 mg L−1 Kan. The transformation efficiency under the optimized conditions was 11.7%. A Southern blot analysis confirmed the integration of the transgene. These results indicated that the transformation efficiency of adult citrus tissues can be enhanced by optimizing the transformation conditions.
KeywordsCitrus Adult tissue Transformation ‘Tarocco’ blood orange
We thank Liwen Bianji, Edanz Editing China (www.liwenbianji.cn/ac) for editing the English text of a draft of this manuscript.
This research was supported by the Science and Technology Innovation Strategy Special Fund of Guangdong Province (2018B020202009), the Earmarked Fund for China Agriculture Research System (CARS-26), the Open Project of Key Laboratory of Horticulture Science for Southern Mountainous Regions, Ministry of Education, and the National Citrus Engineering Research Center.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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