An efficient Agrobacterium -mediated transformation method using hypocotyl as explants for Brassica napus

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An efficient Agrobacterium-mediated transformation method using hypocotyl as explants for Brassica napus Cheng Dai & Yuqing Li & Long Li & Zhuolin Du & Shengli Lin & Xia Tian & Sijia Li & Bao Yang & Wei Yao & Jing Wang & Liang Guo & Shaoping Lu

Received: 25 August 2020 / Accepted: 17 September 2020 # Springer Nature B.V. 2020

Abstract Rapeseed (Brassica napus) is an important oil crop that supplies a considerable amount of global vegetable oil production. Genetic transformation system is important to gene functional analysis and molecular breeding. Here, an efficient Agrobacterium-mediated transformation protocol using hypocotyl of rapeseed as explants is described. To develop this protocol, we compared several essential factors that would affect the transformation efficiency, such as Agrobacterium strains, selection marker genes, and genotypes of rapeseed. Comparison of different Agrobacterium strains showed that the GV3101 had higher transformation efficiency than that of C58C1 and EHA105. HPTII, NPTII, and RePAT were used as selection marker genes in tissue culture. The results showed that the transformation efficiency was 3.7–4.8%, 2.2–22.5%, and 1.6– 5.9% when the hypocotyl of Westar was infected by GV3101 and screened under hygromycin, kanamycin, and basta, respectively. The transformation efficiency of The authors responsible for the distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors are Shaoping Lu and Liang Guo. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11032-020-01174-0) contains supplementary material, which is available to authorized users. C. Dai : Y. Li : L. Li : Z. Du : S. Lin : X. Tian : S. Li : B. Yang : W. Yao : J. Wang : L. Guo (*) : S. Lu (*) National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, China e-mail: [email protected] e-mail: [email protected]

Westar was the highest and ZS11 was the lowest when five different genotypes of rapeseed (Westar, ZS9, ZS11, GY284, and WH3417) were infected by GV3101. Using this protocol, it will take 8–10 weeks to obtain transgenic plants. This protocol has been used to study gene function in several genotypes of rapeseed in our laboratory. These results indicate that it is efficient to obtain transgenic plant of rapeseed using this protocol. Keywords Rapeseed . Transformation method . Hypocotyl . Agrobacterium . Transformation efficiency

Introduction Rapeseed (Brassica napus, AACC, 2n = 38) is one of the most important oil crops and it produces approximately 15% of edible oil globally (Gracka et al. 2016; Yu et al. 2016). It originates from a spontaneous hybridization between Brassica rapa (AA, 2n = 20) and Brassica oleracea (CC, 2n = 18) (Chalhoub et al. 2014; Rahman et al. 2017). Conventional breeding of rapeseed is labor resource intensive and time consuming, which takes eight to ten generations to develop a new variety (Mason and Snowdon 2016).