^*Author for Correspondence.
Institute of Biological Sciences,
Faculty of Science, University of Malaya,
50603 Kuala Lumpur, Malaysia.
Tel: 603-7967 4380.
Email: This email address is being protected from spambots. You need JavaScript enabled to view it.

Key words: Somatic embryogenesis, Plant transformation, Biolistic bombardment, GUS localization, transgenic banana
Abbreviations: 6-BA: 6-Benzyladenine, 2,4-D: 2,4-Dichlorophenoxyacetic Acid, NAA: 1-Naphthaleneacetic acid, 2ip: N⁶-(2isopentenyl)adenine, SCV: settled cell volume, GUS: β-glucuronidase

Abstract.
To obtain stable expression of a foreign gene, it is important to have optimized conditions for plant genetic transformation. Here, a GUS intron-containing gene driven by CaMV 35S promoter was used to optimize the conditions of biolistic transformation of banana immature embryos and also for the study of GUS localization in the transformed cells. The GUS activity detected histochemically and fluorometrically were further analysed by microscopy. This histological study confirmed the results in the histochemical and fluorometric assay, where highest expression of GUS was achieved when the immature embryos bombarded with the helium pressure of 1350 psi and placed at the target distance of 6 cm. The observation of strong GUS staining in the deep layers of the cell structure were produced by higher acceleration pressure and shorter target distance, whereas weak GUS staining in the plant epidermis layer were observed in most lower acceleration pressure and higher target distance. The study of GUS localization on biolistic transformation provided more reliable parameters for transformation. It may also indicate the locality of the foreign gene expression area in the transgenic plant that can provide more understanding of the nature of transgene and its integration.

^*Author for Correspondence.
School of Biological Sciences
Universiti Sains Malaysia
11800 Minden, Penang Malaysia
Tel: 604–6533506; Fax : 604-6565125
Email : This email address is being protected from spambots. You need JavaScript enabled to view it.

Key words: Ganoderma, oil palm, coconut, RAPD, RAMS

Abstract.
Random amplified polymorphic DNA (RAPD) and random amplified microsatellite (RAMS) analyses were used to determine the genetic relatedness within and between Ganoderma boninense isolates from infected oil palm and Ganoderma sp. from coconut stumps from different locations in Malaysia. RAPD analysis using four random primers (5'ACCTGGACAC3', 5'CAGCGACAAG3', 5'AGAGGGCACA3' and 5'TGACGGCGGT3') showed variations of banding patterns within and between the isolates from oil palm and coconut stumps, indicating that they were genetically heterogeneous. There was no specific banding pattern that could differentiate between G. boninense isolates from infected oil palm and Ganoderma sp. from coconut stumps. RAMS analysis using four microsatellite primers, 5'BDB(ACA)₅, 5'DD(CCA)₅, 5'DHB(CGA)₅ and 5'YHY(GT)₅G, also showed variable banding patterns among the isolates from infected oil palm and coconut stumps. However, five common bands i.e. two bands (900 bp and 1200 bp) produced by primer (CGA)₅, one band (1400 bp) by primer (ACA)₅ and two bands (350 bp and 380 bp) by primer (CCA)₅ were shown by all the G. boninense isolates from infected oil palm and Ganoderma sp. from coconut stumps. Dendrograms from cluster analysis based on UPGMA of RAPD and RAMS data showed that G. boninense isolates from infected oil palm and Ganoderma sp. from coconut stumps did not cluster separately into two distinct clusters, but were clustered together, which indicated that both groups of Ganoderma are closely related. The finding that the Ganoderma isolates from coconut stumps are closely related to G. boninense isolates from infected oil palm would have an important bearing in the formulation of disease control measures and replanting procedures, especially in areas where the previous crop was coconut.

^*Author for Correspondence.
Department of Biotechnology
Asian Institute of Medicine
Science and Technology (AIMST)
Amanjaya, 08000, Sungai Petani, Kedah
Tel: 604-4422884; Fax: 604-4422887
Email : This email address is being protected from spambots. You need JavaScript enabled to view it.

Key words: Particle bombardment, Banana, Transient gene expression
Abbreviations: GUS: β–glucuronidase; GFP: Green fluorescent protein, MS: Murashige and Skoog

Abstract.
Physical and biological parameters for DNA delivery into banana cultivar, Rastali (AAB) explants were optimised by monitoring transient gusA and gfp gene expression. Optimisation of the physical factors was carried out under the following conditions; helium pressure (450, 900, 1100, 1350 and 1550psi); distance from stopping plate to target tissue (3, 6, 9 and 12 cm); vacuum pressure (26, 27, 28 and 29 mmHg), number of bombardments (1, 2 and 3 times) per Petri dish and gold microcarrier size (0.6 and 1.0 µm). Distance from rupture disk to macrocarrier and macrocarrier to stopping screen was fixed at 4mm and 11mm, respectively. Two controls were also incorporated i.e. tissues without bombardment and bombardment of microcarrier without DNA. The biological parameters included the explant type (single bud and corm slice), influence of explant sizes (3, 5 and 10 mm), preculture treatment prior bombardment (0, 1, 2, 3 and 4 days), DNA concentrations (0.5, 1.0, 1.5, 2.0 and 2.5 µg) and effect of post-bombardment incubation time (3, 6, 9, 12, 15 and 18 days). Optimised bombardment conditions for single bud was bombarding twice at 1100 psi, 9cm target distance, 28 mmHg, 1 µM gold particle size, 3mm in size range, 1.5 µg DNA per bombardment, three days preculture prior to bombardment and six days post bombardment. For corm slice, the optimised bombardment conditions was bombarding once at a helium pressure of 1100 psi helium pressure, 9cm (gusA gene) or 6cm (gfp gene) target distance, 28 mmHg vacuum pressure and 1 mM gold particle size, 5mm in size range, 1.5 mg DNA per bombardment, one day preculture prior to bombardment and nine days post bombardment. Combinations of optimised physical and biological parameters and an effective selection system were developed which allowed high-efficiency of DNA delivery combined with minimum damage to target banana tissues.

^*Author for Correspondence.
Department of Process and Food Engineering
Faculty of Engineering,
Universiti Putra, Malaysia, 43400 UPM
Serdang, Selangor,Malaysia
Tel: 603 – 89466366; Fax: 603 – 86567123
Email: This email address is being protected from spambots. You need JavaScript enabled to view it.

Key words: anion exchange, dye-ligand, Cibacron Blue 3GA, fluidised bed adsorption, G3PDH

Abstract.
The comparative recovery performance of anion exchange and dye ligand fluidised bed adsorption of intracellular enzyme, glyceraldehydes 3-phosphate dehydrogenase (G3PDH) from unclarified disrupted yeast has been undertaken. The commercially available anion exchanger, Streamline QXL and the kiesleguhr-agarose composite adsorbent, Microsorb K6AX derived with dye-ligand (Cibaron Blue 3GA) were employed in fluidized bed experiments. The adsorbents were evaluated in respect of recovery performance in terms of yield, purity and enzyme specific activity.