Biotechnology-derived food crops are currently being developed in Malaysia mainly for disease resistance and improved post harvest quality. The modern biotechnology approach is adopted because of its potential to overcome constraints faced by conventional breeding techniques. Research on the development of biotechnology-derived papaya, pineapple, chili, passion fruit, and citrus is currently under way. Biotechnology-derived papaya developed for resistance to papaya ringspot virus (PRSV) and improved postharvest qualities is at the field evaluation stage. Pineapple developed for resistance to fruit black heart disorder is also being evaluated for proof-of-concept. Other biotechnology-derived food crops are at early stages of gene cloning and transformation. Activities and products involving biotechnology-derived crops will be fully regulated in the near future under the Malaysian Biosafety Law. At present they are governed only by guidelines formulated by the Genetic Modification Advisory Committee (GMAC), Malaysia. Commercialization of biotechnology-derived crops involves steps that require GMAC approval for all field evaluations and food-safety assessments before the products are placed on the market. Public acceptance of the biotechnology product is another important factor for successful commercialization. Understanding of biotechnology is generally low among Malaysians, which may lead to low acceptance of biotechnology-derived products. Initiatives are being taken by local organizations to improve public awareness and acceptance of biotechnology. Future research on plant biotechnology will focus on the development of nutritionally enhanced biotechnology-derived food crops that can provide more benefits to consumers.
The purpose of this study was to evaluate the effectiveness of using RNA interference in down regulating the expression of 1-aminocyclopropane-1-carboxylic acid oxidase gene in Eksotika papaya. One-month old embryogenic calli were separately transformed with Agrobacterium strain LBA 4404 harbouring the three different RNAi pOpOff2 constructs bearing the 1-aminocyclopropane-1-carboxylic acid oxidase gene. A total of 176 putative transformed lines were produced from 15,000 calli transformed, selected, then regenerated on medium supplemented with kanamycin. Integration and expression of the targeted gene in putatively transformed lines were verified by PCR and real-time RT-PCR. Confined field evaluation of a total of 31 putative transgenic lines planted showed a knockdown expression of the targeted ACO1 and ACO2 genes in 13 lines, which required more than 8 days to achieve the full yellow colour (Index 6). Fruits harvested from lines pRNAiACO2 L2-9 and pRNAiACO1 L2 exhibited about 20 and 14 days extended post-harvest shelf life to reach Index 6, respectively. The total soluble solids contents of the fruits ranged from 11 to 14° Brix, a range similar to fruits from non-transformed, wild type seed-derived plants.
In 2000, agriculture contributed 13% to the national gross domestic product of Malaysia. The country of 23 million people has created a competitive program coordinated by the Ministry of Science, Technology and the Environment, research institutions, and universities to undertake biotechnology research in several areas. Intensified research efforts are under way on oil palm, rubber, rice, papaya, and orchids. Although the most progress has been made in rice and papaya, no transgenic crop is ready for field trials. Nonetheless, preliminary steps have been taken to prepare for the trials, and detailed testing protocols are being developed.
A high survival rate for transformed papaya plants when transferred to the field is useful in the quest for improving the commercial quality traits. We report in this paper an improved rooting method for the production of transformed Malaysian Eksotika papaya with high survival rate when transferred to the field. Shoots were regenerated from embryogenic calli transformed with antisense and RNAi constructs of 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) genes using the Agrobacterium tumefaciens-mediated transformation method. Regenerated transformed shoots, each measuring approximately 3-4 cm in height, were cultured in liquid half-strength Murashige and Skoog (MS) medium or sterile distilled water, and with either perlite or vermiculite supplementation. All the culturing processes were conducted either under sterile or nonsterile condition. The results showed that rooting under sterile condition was better. Shoots cultured in half-strength MS medium supplemented with vermiculite exhibited a 92.5% rooting efficiency while perlite showed 77.5%. The survival rate of the vermiculite-grown transformed papaya plantlets after transfer into soil, contained in polybags, was 94%, and the rate after transfer into the ground was 92%. Morpho-histological analyses revealed that the tap roots were more compact, which might have contributed to the high survival rates of the plantlets.