In vitro regeneration and overexpression of pea DNA helicase 45 (PDH45) gene through Agrobacterium-mediated genetic transformation in oilseed Brassica spp.

Abstract

An efficient genotype independent method for in vitro regeneration and genetic transformation of Brassica spp. viz. Brassica juncea and Brassica napus have been developed for the local varieties using cotyledonary leaf with petioles and hypocotyls as explants. Explants obtained from 6-day old aseptically grown seedlings were inoculated and co-cultivated with Agrobacterium tumefaciens strain EHA105 harbouring a binary vector with PDH45 gene under the regulatory control of 35s promoter and terminator sequences, permitting transformed shoots to be selected on hygromycin containing medium. Well rooted transformed plants were transferred to pots containing soil and after acclimatization the plants were maintained under controlled environmental condition. A total of 9 transformed plants were obtained from BARI Sarisha-8 variety, presumably indicating this protocol is more amenable to genotype independent genetic transformation. Integration and expression of the introduced transgene (PDH45 and hptII) were analysed by polymerase chain reaction (PCR). Factors influencing transformation efficiency include explant age, optical density of Agrobacterium culture for infection, duration of infection and co-cultivation with Agrobacterium were also assessed. Genetic transformation method developed in this study certainly could be utilized for introducing abiotic stress resistance in the local cultivars of oilseed Brassica. Dhaka Univ. J. Biol. Sci. 30(3 CSI): 345-358, 2022 (June)

Efficient in vitro Regeneration of Brassica napus L. Var. BARI Sorisha-18

“…2). According to numerous studies, explants aged between four and six days were shown to provide the best rates of shoot regeneration (Cardoza and Stewart 2004, Mollika et al 2011, Khalil et al 2022. In this study, most shoots are produced from seven to eight-day-old seedlings from both explants, which indicates that the effects of seedling's age on regeneration frequency vary from explant to explant.…”

Establishment of an Efficient in vitro Regeneration Protocol for Brassica campestris L. Var. Tori-7

“…This result indicates that age of seedling affects the callus and shoot regeneration frequency and most range of shoot is produced from 6 days old seedling from both explants. Many researchers have reported that explants excised from 4-6 days old seedlings produced optimal shoot regeneration rates (Cardoza and Stewart 2004, Mollika et al 2011, Khalil et al 2022. It was also narrated that younger explants exhibited greater morphogenic potential than older explants as they might have more metabolically active cells with hormonal and nutritional conditions that are responsible for increased organogenesis.…”

Response of the transcription factor BABY BOOM of Arabidopsis thaliana L. in the formation of embryogenic calluses of cocoa leaves (Theobroma cacao L.)

“…Knowledge of the functionality of genes is essential when it comes to performing Cocoa breeding, which can be achieved through gene overexpression, inactivation, silencing and genetic editing, allowing the integration of agronomically important traits. A good example of success of cocoa breeding is the obtainment of varieties with resistance to Colletotrichum gloeosporioides (Maximova et al, 2006;Khalil et al, 2022). The success of genetic modification depends on the regeneration capacity of transformed plants.…”