Agrobacterium tumefaciens-mediated transformation of drumstick (Moringa oleifera Lam.)

Abstract

Drumstick (Moringa oleifera Lam.) is an industrially important tree species. Due to the availability of an efficient regeneration system, the present study was undertaken to develop a reliable and stable Agrobacterium tumefaciens-mediated transformation system for the genetic improvement of drumstick. A. tumefaciens strain EHA105, harbouring the plasmid pCAMBIA1301 containing the hygromycin phosphotransferase II and ß-glucuronidase genes driven by the CaMV 35S promoter, was used to establish a transformation system in drumstick. The use of vacuum infiltration-assisted Agrobacterium infection (excluding a pre-culture step) and co-cultivation for two days significantly increased the transformation frequency. Genomic integration and transgene expression were confirmed by b-glucuronidase (GUS) assays, polymerase chain reaction (PCR), Southern blot analysis and real-time quantitative PCR (RT-qPCR). This transformation protocol provides a basis for the future development of genetic engineering techniques to improve the performance of drumstick.

Global Transcriptomic Analysis Reveals Differentially Expressed Genes Involved in Embryogenic Callus Induction in Drumstick (Moringa oleifera Lam.)

“…Embryogenic callus induction is therefore seen as a key step in transformation. We have developed an Agrobacterium-mediated transformation system for drumstick [22], but it has not been widely used due to its low transformation efficiency, which is largely due to the difficulty of inducing embryogenic callus formation. Embryogenic callus induction is thus a key bottleneck in drumstick regeneration and hinders its transgenic breeding.…”

An efficient protocol for regenerating shoots from paper mulberry (Broussonetia papyrifera) leaf explants

“…The standard method of paper mulberry proliferation is seed breeding, but the survival rate is low and uniform seeding is not obtained by seed proliferation [ 20 ]. Paper mulberry can also proliferate through root cultivation and cutting propagation [ 21 , 22 ], but the efficiency is relatively low and cannot meet large-scale production needs.…”

Some important aspects in Moringa oleifera Lam. micropropagation

“…Consequently, micropropagation was essential prerequisite of propagation of elite tree. Moringa micropropagation was accomplished using tissues obtained from seedling or mature trees (Islam et al, 2005;Riyathong et al, 2010;Förster et al, 2013;Salem, 2016;Zhang et al, 2017). Vitrification and the associated somaclonal variation are two aspects retarding the application of in vitro techniques as effective vegetative micropropagation tools in moringa (Hassanein et al, 2008;Mirzai et al, 2015;Salem, 2016;Hassanein et al, 2018).…”

“…Under stress conditions, SOD plays its role by catalyzing the dismutation of excess O 2 − into O 2 and H 2 O 2 . Toxic H 2 O 2 is further catalyzed by CAT, POX and other enzymes to form H 2 O and O 2 (Ueda et al, 2013;Rout et al, 2013;Zhang et al, 2017).…”