Transgenic Wucai (Brassica campestris L.) produced via Agrobacterium-mediated anther transformation in planta
We developed a novel Agrobacterium-mediated anther transformation for Wucai in planta, and in this procedure, the male germ line was the predominant target.
Wucai (Brassica campestris L.), a variant of non-heading Chinese cabbage, is widely cultured in China and only improved by classic breeding methods. Here, a novel and efficient in planta Agrobacterium-mediated anther transformation method is developed based on the optimization of several factors that affect anther transformation. After optimization, transformation with the manual pollination application led to increased transient GUS expression in anthers (reaching 91.59%) and the transformation efficacies in planta (0.59–1.56% for four commercial cultivars). The stable integration and inheritance of the transgenes were further examined by molecular and genetic analyses. Three T2 transgenic lines presented a segregation ratio of 3:1, which was consistent with the Mendelian feature of a single dominant gene. In addition, the GUS histochemical assay and genetic crossing analysis revealed that the male germ line was the predominant target in this transformation. This optimized transformation system could provide a useful tool for both the improvement of cultivar qualities and investigation of functional genes in Wucai.
KeywordsAnther-drop Cytological observation Genetic transformation In planta Non-heading Chinese cabbage
6-Benzyl amino purine
Cytoplasmic male sterility
Environmental scanning electron microscopy
Morpholino ethane sulfonic acid
Murashige and Skoog medium
The National Natural Science Foundation of China (31801853); National Key R & D Program of China (2017YFD0101803); Provincial Science and Technology Major Project of Anhui (17030701013); and Major (KJ2017ZD15) and Key (KJ2017A153) Projects of Natural Science Research Funds in Support of Colleges supported this work.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interests.
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