Doing the “Floral Dip” (DIY Plant Genetic Engineering)

1009345066 4bfdc688afMaking Transgenic Plants Without Using Tissue Culture

Since the first successful genetic transformation of plants using Agrobacterium in 1983, many methods for transferring foreign genes into plants have been developed, including the so-called “gene guns”. (A good overview is presented in Ref 1 below.)

Many, if not most, of these methods rely on plant tissue culture to regenerate transgenic plants.

However, plant tissue culture is typically quite time consuming and labor intensive, and it can introduce abnormalities in regenerated plants due to somaclonal variation.

Therefore, other methods to transform plants were investigated, including the inoculation of Agrobacterium into intact plants.

This so-called in planta genetic transformation was first successfully achieved in 1994, as reported here.

Many variations on this in planta approach were published during the 1990’s. Most involved the vacuum infiltration of a suspension of Agrobacterium into the plant.

But an in planta method that really seemed to catch on did not involve vacuum infiltration. It was called the “floral dip” method, published by Clough and Bent in 1998. (Please see Ref 2 below.)

Briefly, to inoculate plants, developing flowers of intact Arabidopsis plants were simply dipped into a suspension of Agrobacterium, and the plants were allowed to produce seeds normally. The seeds were then germinated on agar-based selection media (containing herbicide, e.g.), and the successfully transformed plant seedlings were identified and propagated.

A complete description of the protocol can be found at, where else, floraldip.com.

Since 1998, this method has been refined and modified, including the floral inoculation method (Please see Ref 1 below, for example.) It has also been used to genetically transform a variety of plant species other than Arabidopsis thaliana.

References

1. Narusaka, Y., et al. (2012) “Methods to transfer foreign genes to plants.” IN: Transgenic Plants – Advances and Limitations, Yelda Ozden Çiftçi (Ed.), ISBN: 978-953-51-0181-9, In-Tech, pp. 173-188. (Full Text)

2. Clough, S.J. and A.F. Bent (1998) “Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana.” Plant Journal, Vol. 6, pp. 735-743. (PDF)

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