Abstract
Dendrobium Broga Giant (Dendrobium Bobby Messina × Dendrobium Superbiens) is a newly introducing hybrid orchid in Malaysia. Determination of target explants potentiality is very important before transformation studies. Bacterial chemotaxis and Transient gusA expression studies were conducted on Dendrobium Broga Giant orchid’s PLBs to evaluate the capability of the bacterial-PLBs interaction during transformation process. Agrobacterium tumefaciens showed positive chemotactic response and their attachment to the PLBs whereas E. coli response negatively. Mild wounded PLBs displayed higher Agobacterium motility compared to other treatments. Mild wounded PLBs immersed for 15 minutes on the Agrobacterium suspension (OD600 nm 0.8) and co-cultivated for 2 days in co-cultivation media supplemented with 200μM acetosyringone showed highest transient gusA gene expression in Dendrobium Broga Giant orchid. Hence, we concluded that positive chemotactic responses and transient gusA gene expression ability indicate the Dendrobium Broga Giant orchid PLBs are a potential target explants for Agrobacterium –mediated genetic transformation studies
Highlights
Orchid is an important group of ornamental plants comprising of several thousand species and hybrid
Bacterial swarming was able to be seen towards the naked eye which was authorized to measure the chemotaxis response of Agrobacterium tumefaciens
The maximum (1.13) ratio of chemotaxis movement was recorded from 48 hours after inoculation and the minimum (1.07) ratio was found from 24 hours after inoculations (Fig. 3)
Summary
Orchid is an important group of ornamental plants comprising of several thousand species and hybrid. Transient gusA expression is an indicator of gene transfer and expression efficiency for the optimization of Agrobacterium-mediated transformation protocols which is coding for the enzyme β-glucoronidase (GUS) used as a reporter gene to identify the transgenic plants Several factors such as types of wounding, bacterial immersion period, co-cultivation period, density of Agrobacterium (OD600nm) and acetosyringone concentrations in cocultivation medium which are essential for transformation studies. Attachment of Agrobacterium to plant cells can be observed during chemotaxis and histochemical GUS assays, the specificity of the cell-cell contact would be preferably confirmed by a transient gusA measurement of the binding aptitude of attachment able to bacteria This present study evaluates the preliminary assessments on Agrobacteriummediated transformation of Dendrobium Broga Giant’s PLBs
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