Establishment of A Simple and Efficient Agrobacterium-mediated Genetic Transformation System to Chinese Cabbage (Brassica rapa L. ssp. pekinensis)

Li, Xiaonan; Li, Haiyan; Zhao, Yuzhu; Zong, Peixuan; Zhan, Zongxiang; Piao, Zhongyun

doi:10.1016/j.hpj.2021.01.006

Cited by 25 publications

(22 citation statements)

References 42 publications

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“…During the entire process, the cotyledon explants passed through several stages, which hinder their Agrobacterium-mediated regeneration ability as observed by Karthikeyan et al (2011) in rice transformations. The transformed regenerated plantlets were shifted to subsequent hardening, and a transformation efficiency of 14% was achieved (Table 1), which is significantly higher than earlier reports (Jun et al, 1995;Zhang et al, 2000;Lee et al, 2004;Ku et al, 2006;Min et al, 2007;Konagaya et al, 2013;Baskar et al, 2016;Liu et al, 2018;Li et al, 2021). Various transformation efficiencies (0.6-10.38%) in B. rapa L. ssp.…”

Section: Discussionmentioning

confidence: 74%

“…pekinensis) (Liu et al, 2018). Li et al (2021) used 25 mg/L hygromycin B to select transgenic plants from cotyledon explants of B. rapa. Slightly increasing the concentration of selection agents from the minimum inhibitory concentration by passing one more cycle with a week interval has greatly contributed in recovery of trueto-type of transgenic plants (Sivanandhan et al, 2015).…”

Section: Discussionmentioning

confidence: 99%

“…pekinensis ) ( Liu et al, 2018 ). Li et al (2021) used 25 mg/L hygromycin B to select transgenic plants from cotyledon explants of B. rapa .…”

Section: Discussionmentioning

confidence: 99%

“…Despite this recalcitrance, genetic development through Agrobacterium -mediated transformation is a powerful way to improve B. rapa traits. There are limited available reports on the Agrobacterium -mediated transformation of Chinese cabbage ( Jun et al, 1995 ; Zhang et al, 2000 ; Lee et al, 2004 ; Ku et al, 2006 ; Min et al, 2007 ; Konagaya et al, 2013 ; Baskar et al, 2016 ; Liu et al, 2018 ; Li et al, 2021 ). Jun et al (1995) reported 3% of transformation efficiency from cotyledon explants of Chinese cabbage ( B. campestris L. ssp.…”

Section: Introductionmentioning

confidence: 99%

“…pekinensis ) and used kanamycin as a selection agent. Li et al (2021) attained 10.83% of transformation efficiency in cotyledon explants of Chinese cabbage ( B. rapa L. ssp. pekinensis ) using hygromycin as a selection agent.…”

Section: Introductionmentioning

confidence: 99%

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L-Cysteine Increases the Transformation Efficiency of Chinese Cabbage (Brassica rapa ssp. pekinensis)

et al. 2021

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Successful Agrobacterium-mediated transformations of Chinese cabbage have been limited owing to the plant’s recalcitrant nature, genomic background and explant necrosis upon infection, which hinders the transfer of T-DNA region into the Chinese cabbage. Consequently, in the current experiment, a stable Agrobacterium tumefaciens-mediated transformation method for Chinese cabbage cv. Kenshin established by employing important anti-oxidants in the co-cultivation and subsequent regeneration media. Four-day-old in vitro derived cotyledon explants were infected with A. tumefaciens strain GV3101 harboring the vector pCAMIBA1303. Cotyledon explants exposed to an Agrobacterium suspension (OD600 of approximately 0.6) for 10 min and then incubated for 3 days co-cultivation in Murashige and Skoog medium containing an L-cysteine + AgNO3 combination exhibited the highest β-glucuronidase (GUS) expression (94%) and explant regeneration efficiency (76%). After 3 days, the cotyledon explants were subjected to three selection cycles with gradually increasing hygromycin B concentrations (10 to 12 mg/L). The incorporation and expression of hptII in T0 transformed plants were verified by polymerase chain reaction and Southern blot analyses. These transgenic plants (T0) were fertile and morphologically normal. Using the present protocol, a successful transformation efficiency of 14% was achieved, and this protocol can be applied for genome editing and functional studies to improve Chinese cabbage traits.

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