In vitro Regeneration and Agrobacterium-mediated Genetic Transformation of Local Varieties of Mungbean (Vigna radiata (L). Wilczek)

Abstract

An efficient Agrobacterium-mediated transformation compatible in vitro regeneration protocol was developed for two important varieties of mungbean (Vigna radiata (L.) Wilczek) cultivated in Bangladesh, namely Binamoog-5 and BARI Mung-6. Two different zygotic embryo derived explants, such as cotyledonary node (CN) and cotyledon attached decapitated embryo (CADE) were used for direct organogenesis of shoot. MS supplemented with 4.0 μM BAP was found to be the best for the development of highest number of multiple shoots from CADE in both the varieties of mungbean. While in case CN the best shoot formation was achieved on MS containing 4.0 μM BAP and 0.5 μM NAA in both varieties. Half strength of MS with 2.0 μM IBA was found to be most effective for producing healthy root from regenerated shoots. Following root induction, the in vitro raised plantlets were successfully transplanted to soil for their establishment. Considering overall responses, genetic transformation efficiency was found to be better with CADE explant using Agrobacterium tumefaciens strain LBA4404 harboring the binary plasmid pBI121 conferring GUS and nptII genes. Different factors influencing transformation was optimized during this study. Selection of transformed shoots was carried out by gradually increasing the concentration of kanamycin and such transformed shoots were eventually selected using 200 mg/l kanamycin. Stable expression of the GUS gene was detected in various parts of regenerated transformed plantlets. Transformed shoots were rooted on half strength MS containing 2.0 μM IBA and 100 mg/l ticarcillin. Rooted transformed plantlets were successfully transferred to soil. Stable integration of GUS and nptII genes in the putative transformed shoots was confirmed through PCR analysis.
 Plant Tissue Cult. & Biotech. 29(1): 81-97, 2019 (June)

Elicitation of the in vitro Cultures of Selected Varieties of Vigna radiata L. With Zinc Oxide and Copper Oxide Nanoparticles for Enhanced Phytochemicals Production

“…A moderate effect of BA in the production of multiple shoots has been previously reported in the mung bean by Vats et al (2014). There are also some other reports of tissue culture of mung bean varieties where BAP produced better results (Hoque et al, 2007;Bhajan et al, 2019). Some in vivo studies also show that foliar application or spray of BAP gave higher per hectare yields of mung beans (Sarker et al, 2021).…”

In Planta Genetic Transformation of Mungbean (Vigna radiata (L.) Wilczek) with Marker Gene

“…Moreover, due to the antibiotic selection the success rate droped drastically and in case to shoot regeneration and viable root formation in vitro. Mostly the success rate varies between 1 and 2% but the rate droped further during the acclimatization stage of the in vitro regenerated plantlets at environmental conditions (Bhajan et al 2019). Therefore, in planta transformation could be an alternative option to produce healthy transgenic mungbean plants.…”

“…Though it has high demand the production rate is far below. As a result, Bangladesh has to import a large amount of mungbean from its neighbouring countries (Bhajan et al 2019).…”

“…However, a reproducible and efficient in vitro regeneration protocol is a pre-requisite of plant genetic transformation (Mohanty and Sagare 2015). Though successful in vitro regeneration of mungbean plantlets were reported from various explants, for instance, cotyledonary nodes (Mohanty and Sagare 2015, Vats et al 2014, Himabindu et al 2014, cotyledon, hypocotyl, root and shoot tip (Khatun et al 2008), double cotyledonary node (Yadav et al 2010), decapitated mature embryo (Hoque et al 2007), cotyledon attached decapitated embryo (Bhajan et al 2019) it is still considered as a recalcitrant plant towards tissue culture particularly in case of developing healthy and viable root system. Development of Agrobacterium-mediated genetic transformation protocol has also achieved very little success in mungbean as it is not amenable towards genetic modification (Vir et al 2016, Dewir et al 2016.…”

“…Till date, very little success has been achieved towards the genetic modification of mungbean and most of them are confined with reporter gene (e.g. gus) (Bhajan et al 2019, Yadav et al 2012, Tazeen and Mirza 2004, Jaiwal et al 2001. There are some reports of successful development of transgenic mungbean with alien gene (Baloda and Madanpotra 2017, Kumar et al 2017, Sahoo et al 2016, Vijayan and Kirti 2012, Saini et al 2007 but the success rate is reported to be between 1 and 2%.…”