Expression of Bacillus thuringiensis insecticidal protein gene in transgenic oil palm

Lee, Mei Phing, and Yuen, Li Huey, and Ruslan Abdullah, (2006) Expression of Bacillus thuringiensis insecticidal protein gene in transgenic oil palm. Electronic Journal of Biotechnology, 9 (2). ISSN 07173458

Abstract

Oil palm (Elaeis guineensis Jacq.) is the most sought-after edible oil in the world. Advances in agricultural biotechnology, in particular through gene manipulation, have seen the monopoly enjoyed by palm oil gradually eroded by genetically modified oil-bearing crops such as soybean, sunflower seed, and rapeseed. Thus, efforts are being undertaken to increase oil palm yield to meet the ever-increasing global demands and also to broaden its spectrum of uses. Conventional oil palm breeding is inefficient and time-consuming. However, current breeding programme combines both conventional and modern biotechnology approaches such as the use of tissue culture technique, genetic transformation and marker-aided selection. Although many studies have been done on genetic manipulation of oil palm, but publications are limited. This is probably due to the high commercial implication tagged to the crop. It may also be due to the crops’ own physiological characteristics such as long generation cycle. Over the past 10 years, our group has been actively involved in genetic engineering of oil palm. Special emphasis was placed initially on developing techniques for genes transfer into oil palm, both using direct gene transfer and Agrobacterium-mediated approaches. Both techniques were then used to transfer useful genes such as cowpea trypsin inhibitor (CpTI) and Bacillus thuringiensis (Bt) crystal insecticidal protein genes, chitinases and more recently, genes for biosynthesis of bioplastics. In most gene manipulation studies, the main bottleneck lies in assessing the functionality of transgenes in putative transformants. Very often, these could not be clearly demonstrated. Here, several expression vectors carrying CryIA(b) gene were constructed and transformed into oil palm. Successful transfer of the CryIA(b) gene into oil palm was clearly demonstrated followed by analysis on the transgenes functionality in its new environment. The approach used combines both RT-PCR and Southern Blot analysis, and was proven effective to elucidate the presence and functionality of transgenes. In addition, it also provides a rapid technique to screen for putative transformants that proved very difficult in gene manipulation of monocots, especially those having slow growing rate in vitro such as oil palm.

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Abstract