Challenges of In Vitro and In Vivo Agrobacterium-Mediated Genetic Transformation in Soybean

Abstract

Agrobacterium tumefaciens-mediated genetic transformation of plants is a natural process. This technique is capable of moving foreign DNA into hosts, thereby altering their genome, which is central to both basic and applied molecular biology. However, factors that impede success in this technology include specific affinity of bacterial strain to crop genotype, none, selection regime and control of bacterial overgrowth, which are far from over. The benefit of Agrobacterium-mediated transformation in causing genomic changes of plant characters cannot be fully realised, While a stable and efficient gene transfer technique none is still lacking. Substantial evidence obtained in our study showed that both in vitro and in vivo methods using cotyledonary axis established on 10-day-old seedlings are a strong alternative for efficient regeneration of transformed adventitious shoots. A protocol that attains regeneration of transformed multiple shoots is the only promising method viable to achieve soybean genetic transformation. High shoot regeneration of 60.0%, 63.3% and 76.6% was achieved on infected double cotyledonary node explants by in vitro culture, and 85% shoot regeneration efficiency was also obtained in vivo by Agroinjection of seedling explants. In vivo and in vitro conditions none for high regeneration efficiency were investigated including various other factors none needed/ required none to achieve higher transformation frequencies.

Genetic Transformation in Agro-Economically Important Legumes

“…It is also referred to as biolistics (short for biological ballistics) and involves the direct transfer of DNA-coated particles into semi-permeabilised host cells using high-speed propulsion [68]. It was used over 2 decades ago to develop the first transgenic crop called Roundup Ready and has continuously been used to transform various other plants [69]. Unlike Agrobacterium-mediated genetic transformation, this technique can be used to transfer transgenes to various host tissues, to transform the chloroplast genome and is said to have a broader host range [60].…”

“…Another miscellaneous method used in the direct transfer of DNA to plants is electroporation-mediated genetic transformation, which employs the uptake of DNA through a semi-permeable plasma membrane by plant cells and protoplasts using an electric pulse [70]. Another method, silicon carbide fibres also known as whiskers, involves the treatment of explant material in a buffer solution that consists of DNA and silicon carbide fibres [68,69]. Although it requires no complex or expensive equipment, the use of this method carries a danger posed by the fibres on human health, and thus requires careful handling by experienced personnel [69].…”

The Role of Plant Genotype, Culture Medium andAgrobacteriumon Soybean Plantlets Regeneration during Genetic Transformation

“…The genetically modified soybeans range from metabolically engineered plants such as those exhibiting increased oleic acid as well as the herbicide-tolerant (HT) cultivars [26,27]. Soybean transformation led to production of elite cultivars, increased gene pool, plants with improved secondary metabolites and production of disease free plants, especially regenerated under aseptic culture conditions [2,28]. As the attempts in soybean transformation progresses, procedures must focus on transgenic plant production exhibiting tolerance to abiotic stress factors.…”

“…However, the vegetative and reproductive stages of soybean continuously show high sensitivity to biotic and abiotic stress constraints. Yield quality and quantity of this crop is severely affected by high temperatures, chilling, waterlogging and water deficit stress [2]. Furthermore, tools such as genetic engineering, aimed at improving the growth characteristics of this crop are also negatively influenced by several factors like genotype specificity and co-cultivation challenges discussed in the above sections.…”

“…The less significant role of soybean in these regions may be due to the poor growth conditions. The growth and productivity of this crop has been adversely affected by the biotic and abiotic stress factors [2]. Even though it has the potential to become a major crop in less cultivated regions (Africa and Europe) because of its many uses (as feed, food, etc.…”