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Citrus Cell Suspension Culture Establishment, Maintenance, Efficient Transformation and Regeneration to Complete Transgenic Plant

文献类型： 外文期刊

作者： Moniruzzaman, M. 1 ; Zhong, Yun 1 ; Huang, Zhifeng 1 ; Yan, Huaxue 2 ; Yuanda, Lv 3 ; Jiang, Bo 1 ; Zhong, Guangyan 1 ;

作者机构： 1.Guangdong Acad Agr Sci, Inst Fruit Tree Res, Guangzhou 510640, Peoples R China

2.Minist Agr, Key Lab South Subtrop Fruit Biol & Genet Resource, Guangzhou 510640, Peoples R China

3.Guangdong Prov Key Lab Trop & Subtrop Fruit Tree, Guangzhou 510640, Peoples R China

关键词： cell suspension; Citrus; tissue culture; de novo organogenesis; genetic engineering

期刊名称：PLANTS-BASEL （ 影响因子：2.762； ）

ISSN：

年卷期： 2021 年 10 卷 4 期

页码：

收录情况： SCI

摘要： Agrobacterium-mediated transformation of epicotyl segment has been used in Citrus transgenic studies. The approach suffers, however, from limitations such as occasionally seed unavailability, the low transformation efficiency of juvenile tissues and the high frequency of chimeric plants. Therefore, a suspension cell culture system was established and used to generate transgenic plants in this study to overcome the shortcomings. The embryonic calli were successfully developed from undeveloped ovules of the three cultivars used in this study, "Sweet orange"-Egyptian cultivar (Citrus sinensis), "Shatangju" (Citrus reticulata) and "W. Murcott" (Citrus reticulata), on three different solid media. Effects of media, genotypes and ages of ovules on the induction of embryonic calli were also investigated. The result showed that the ovules' age interferes with the callus production more significantly than media and genotypes. The 8 to 10 week-old ovules were found to be the best materials. A cell suspension culture system was established in an H+H liquid medium. Transgenic plants were obtained from Agrobacterium-mediated transformation of cell suspension as long as eight weeks subculture intervals. A high transformation rate (similar to 35%) was achieved by using our systems, confirming BASTA selection and later on by PCR confirmation. The results demonstrated that transformation of cell suspension should be more useful for the generation of non-chimeric transgenic Citrus plants. It was also shown that our cell suspension culture procedure was efficient in maintaining the vigor and regeneration potential of the cells.

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