Efficient Agrobacterium tumefaciens-mediated genetic transformation of Aloe vera

Genetic transformation of plants has emerged as a core research tool for the functional characterization of genes and cultivar improvement in the field of plant biology. Effectually, high transformation efficiency is a prerequisite for accomplishing the stated targets. Aloe vera, a tropical cactus plant in distinction to the family Liliaceae, has seized a long history of secular acceptance as a therapeutic agent and is reasonably the most popular herbal remedy these days. However, the genetic transformation efficiency of Aloe vera is relatively low and seldom reported formerly. This study aims to optimize Agrobacterium-mediated transformation in Aloe vera by refining several parameters like explant selection, the extent of explant injury during infection, Agrobacterium concentration, co-cultivation pH, and duration and desiccation of plant tissue to improve the infection efficiency. The results showed that nearly 83% transient transformation efficiency was attained by suspending the Agrobacterium tumefaciens (AGL1 strain) cells at a concentration of OD600: 0.4 in co-cultivation media at pH-5.6 to infect the stem of aloe by desiccation, followed by 3 days of co-cultivation. Desiccation during infection had proved to enhance T-DNA delivery, whereas a higher extent of explant injury was found to curtail the infection efficiency. The molecular analysis of the putative transformants was carried out by following GUS histochemical assay and PCR. Thereupon, Southern blotting was used to substantiate the authentication of positive transgenic plants and analyze the copy number of the hptII gene in the Aloe vera genome. A transformation efficiency of over 2% was obtained, which is higher than the previous report. This study bestows an improved Agrobacterium-mediated transformation protocol based on desiccation in Aloe vera, which might help in facilitating various functional genomic studies and gene regulation in the aloe plant, eventually allowing the genetic modification of aloe species in an effective manner. Key messageThis research deliberates a meliorated Agrobacterium-mediated transformation methodology for Aloe vera, which may facilitate various gene functional investigations and bioengineering of aloe species in an effective salutary way.

Automated summary

This study aims to optimize Agrobacterium-mediated transformation in Aloe vera by refining several parameters to improve the infection efficiency. The results showed that an improved transformation efficiency of over 2% was obtained.