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

Abstract

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.

Effective micropropagation of kale (Brassica oleracea convar. Acephala var. sabellica): one of the most important representatives of cruciferous crops

“…Comparing the substrata used for acclimatization of in vitro-derived kale microplantlets, the highest survival rate result from a mixture of soil and perlite (1:1). Successful acclimatization in a mixture containing perlite was also reported for cabbage (soil:perlite, 3:1) and Chinese cabbage (perlite alone) (Gerszberg et al 2015;Sivanandhan et al 2019). Perlite is well-known for soil aeration and water absorption (Ilahi and Ahmad 2017), thus mixture with perlite is the easiest way to provide optimal conditions for microplantlets.…”

Overexpression of rice acyl-CoA-binding protein OsACBP5 protects Brassica napus against seedling infection by fungal phytopathogens

“…Moreover, the high transformation efficiencies for B. napus-RC plants is a substantial improvement compared to recent studies, which reported transformation efficiencies of 1.4-3% (Min et al 2007), 2.89-5% (Lee et al 2004), 1.2% (Konagaya et al 2013), 15% (Baskar et al 2016) from hypocotyl explants and 10.83% (Li et al 2021) and 14% (Sivanandhan et al 2021) from explants from cotyledons of Brassica rapa L. ssp. pekinensis.…”

“…2016) from hypocotyl explants and 10.83% (Li et al . 2021) and 14% (Sivanandhan et al . 2021) from explants from cotyledons of Brassica rapa L. ssp .…”

Biocontrol potential of cell-free supernatant of Paenibacillus chitinolyticus against Plasmodiophora brassicae in two important Brassica species

“…Chinese cabbage (Brassica rapa L.) is one of the most important green leafy vegetables in the family Brassicaceae with increasing popularity in the world (Sivanandhan et al, 2021;Vanjildorj et al, 2009). Rapeseed (Brassica napus L.) is also one of the most widely grown crops of Brassicas in the world and is the second most important crop after cereal in the Czech Republic (Anonymous, 2018).…”