Genetic Transformation ofMetroxylon sagu(Rottb.) Cultures viaAgrobacterium-Mediated and Particle Bombardment

Abstract

Sago palm (Metroxylon sagu) is a perennial plant native to Southeast Asia and exploited mainly for the starch content in its trunk. Genetic improvement of sago palm is extremely slow when compared to other annual starch crops. Urgent attention is needed to improve the sago palm planting material and can be achieved through nonconventional methods. We have previously developed a tissue culture method for sago palm, which is used to provide the planting materials and to develop a genetic transformation procedure. Here, we report the genetic transformation of sago embryonic callus derived from suspension culture using Agrobacterium tumefaciens and gene gun systems. The transformed embryoids cells were selected against Basta (concentration 10 to 30 mg/L). Evidence of foreign genes integration and function of the bar and gus genes were verified via gene specific PCR amplification, gus staining, and dot blot analysis. This study showed that the embryogenic callus was the most suitable material for transformation as compared to the fine callus, embryoid stage, and initiated shoots. The gene gun transformation showed higher transformation efficiency than the ones transformed using Agrobacterium when targets were bombarded once or twice using 280 psi of helium pressure at 6 to 8 cm distance.

Adaptive Evolution and Addressing the Relevance for Genetic Improvement of Sago Palm Commodity

“…Genetic improvement of sago palm may use transformation agrobacterium-mediated and particle bombardment. Successfully introgression bar and gus gene into sago palm genome [65]. The embryogenic callus was the most appropriate transformation material compared to the via callus, the embryoid stage and the shoots initiated by using Agrobacteriummediated.…”

Kloning dan karakterisasi daerah promoter gen penyandi ADP glucose pyrophosphorylase dari Metroxylon sagu rendemen pati-tinggi dan -rendah [Cloning and characterization of promoter region of ADP glucose pyrophosphorylase-encoding gene from Metroxylon sagu with high- and low-starch content]

“…Sementara dalam upaya mengembangkan marka untuk rendemen pati tinggi, Budiani et al, (2014) mengidentifikasi perbedaan pola RAPD antara sagu rendemen pati tinggi dengan sagu rendemen pati rendah. Dalam bidang molekuler, Ibrahim et al, (2014) menguji efisiensi beberapa metode transformasi pada tanaman ini. Dari konstruksi pustaka cDNA dan analisis EST yang dilakukan, Wee & Roslan (2012), mengidentifikasi transkrip yang terlibat dalam metabolisme primer dan toleran stres.…”

“…Riset terkait biosintesis pati serta cara memodifikasinya telah banyak dilaporkan pada berbagai tanaman terutama pada padi dan gandum (Sestili et al, 2009;Jeon et al, 2010;Slade et al, 2012;Fasahat et al, 2014). Menurut Masouleh et al, (2012) ada tujuh kelompok enzim yang berperan langsung maupun tidak langsung pada produksi granul pati, yaitu AGP, granule bound starch synthase (GBSS), starch synthases (SS), branching enzyme (BE), debranching enzyme (DBE), starch phosphorylase (PHO) dan glucose 6-phosphate translocator (GPT).…”

“…Komposisi pati tidak semata-mata dipengaruhi oleh aktivitas AGP, tetapi merupakan hasil akhir dari keseluruhan tahapan biosintesis pati. Over ekspresi AGP pada tanaman gandum, padi dan jagung dilaporkan meningkatkan jumlah biji dan produksi, yang membuktikan bahwa AGP menentukan akumulasi pati secara keseluruhan (Geigenberger, 2011). Budiani et al, (2015) telah mempelajari perbedaan ekspresi AGP pada tanaman sagu rendemen pati tinggi dengan pada tanaman sagu rendemen pati rendah, dan hasilnya menunjukkan bahwa tanaman sagu rendemen pati tinggi mengekspresikan AGP lebih tinggi dibandingkan tanaman sagu rendemen pati rendah.…”

Molecular And 3D-Structural Characterization Of Fructose-1,6-Bisphosphate Aldolase Derived From Metroxylon Sagu

“…However, in the post-genomic era, sago molecular biology remains unknown which hampers further improvement of starch production and to understand the underlying mechanism of biotic and abiotic stress responses of sago palm. Nevertheless, recent publications in sago palm molecular biology includes characterization of stress-related genes 4,5 , development of expressed sequence tags of young leaves 6 and the advancement of gene transformation method in sago palm 7 . Fructose 1,6-bisphosphate aldolase (EC 4.1.2.13; FBAld) is an enzyme that catalyzes a reversible reaction of glyceraldehyde-3-phosphate (G3P) and dihydroxyacetone phosphate (DHAP) via Schiff base reaction that is crucial for both glycolysis/gluconeogenesis and the pentose phosphate pathways 8 .…”