Sago thenar, a liliopsid, is one of the most of import economic species and now grown commercially in Malaysia, Indonesia, Philippines and Papua New Guinea for production of sago amylum and / or transition to animal nutrient or fuel ethyl alcohol. In many states South East Asia, except Irian Jaya, M. sagu is chiefly found in semi-cultivated bases. Irian Jaya has approximately 6 million hour angle of M. sagu. The bases of good quality M. sagu can be rather big. Papua New Guinea has an estimated 1 million hour angle of wild and 20 000 hour angle of semi-cultivated M. sagu. M. sagu is besides found in Guam, Palau, Nakuoro, Kosrae, and Jaluit, Marshall Islands most likely the consequence of human debut ( Mc Clatchey et al. 2006 )
Sago thenar is a harvest that can bring forth amylum in a whole batch from one tree. It is a robust liliopsid as it can turn in dirts where merely few workss can last such as the waterlogged, immature, acidic and deep peat of Sarawak. This boggy peat environment has covered 12 % of Sarawak ‘s entire land country ( Tie and Lim, 1977 ) and based on this fact, the Sarawak Government has focused their attempt to develop the sago industry through the Department of Agriculture ( DOA ) , Sarawak in 1982.
As an early consequence, a commercial sago plantation was developed in Mukah in 1987 by the Sarawak Land Custody and Development authorization ( LCDA ) every bit good as a Crop Research and Application Unit ( CRAUN ) in 1993 to set about more intensive research and development on sago ( Jong, 1995 ) . One of the research countries conducted by CRAUN is sago thenar tissue civilization. Currently CRAUN ‘s tissue civilization ( TC ) research lab has been developing in vitro extension system in order to get by with the plantation demands where the clonal has being planted on a big graduated table.
However the long ripening period, variable amylum quality and entire deficiency of selected quality assortments as seting stuff are the major issues that need to be overcome by familial betterment of sago thenar. In order to better the bing thenar feature in a short infinite of clip, familial transmutation holds great possible. Sago thenar transformed with new cistrons that is coding for characters such as high output, amylum quality, early adulthood, etc. will be ensuing in the production of improved sago thenar for commercial planting.
Conventional works genteelness has succeeded in bring forthing a broad assortment of commercial workss and harvests with a scope of of import agronomic traits. It has succeeded in change overing a Mexican grass into corn and Middle East grass into wheat. However, it is to a big extent a haphazard procedure, uniting big parts of parental genomes in a instead uncontrolled manner, although this is presently being improved due to the modern technique of marker assisted genteelness. Genetic technology, on the other manus, allows scientists to reassign really specific cistrons into workss, ensuing in the debut of one or more defined traits into a peculiar familial background. This procedure is called familial transmutation and the cistrons involved are expressed to organize a protein responsible for the peculiar trait. The traits involved include weedkiller and drought tolerance, and opposition to viral, bacterial and fungous pathogens every bit good as to herbivorous insects. The added advantage is that the transferred cistron ( s ) , or transgene ( s ) , can come from any being every bit long as its look is compatible with its new host ( Thomson ) .
Genetics transmutation or familial technology engineerings have been widely used for betterment of monocotyledonous harvests such as corn, rice and oil thenar. The betterment of the qualities of these harvests through familial technology has stimulated and so frequently widened their commercialisation chances ( Ruslan et al. , 1997 ) . Sago thenar has one of the of import requirement in the production of transformed or transgenic works ; and that is the ability of its cells to turn, divide and finally renew into works in vitro. Normally during its recovery, wounded workss like sago thenar signifier a mass of uniform tissue known as callosity. The callosity, in bend, can be induced by endocrines in vitro to bring forth shoots in vitro and can besides be used as a mark for familial transmutation utilizing either the biolistic gun or Agrobacterium-mediated methods.
At present, the most routinely and widely used method to obtain transgenic workss is by the usage of Agrobacterium-mediated method. Once this method was less successful with liliopsids but more late it has been possible to transform liliopsids utilizing Agrobacterium-mediated method, for harvests such as rice, maize, yam and palm oil ( Ming Cheng et al. , 2004 ) .
There is an pressing demand to better the sago thenar for plantations through non-conventional methods and it is seasonably to develop a process for familial betterment of sago thenar. However presently there are a few undertakings, conducted in CRAUN and UNIMAS, looking at insulating the cistrons involved in assorted biochemical and physiological procedures in sago thenar. These cistrons one time isolated and characterized, can so be used to better the sago thenar. Ultimately, thenars can be produced with enhanced features such as improved output and amylum quality. Therefore the constitution of a transmutation protocol in sago thenar is indispensable in order to reintroduce the altered cistrons or other fresh cistron sequences into sago genome.
In this survey, saloon and Guam cistrons, will be used as marker cistrons to bespeak the success of the transmutation system. Two attacks were tested in this research undertaking which are Agrobacterium-mediated and microprojectile barrage. Although familial transmutation on monocotyledonous workss is normally known to hold a low-efficiency ( Willmink et Al, 1993 ) , both of these methods were proven to hold succeeded in corn, wheat, rice and oil thenar.
One of the factors that can increase the transmutation efficiency is the technique of effectual choice. Choice after the transmutation is of import to suppress growing of the untransformed cells and to enable transformants to last and renew into complete transgenic workss. Or else, a bulk of nontransformants will rule the civilization and bring forthing Chimera workss.
In works familial technology, it is of import to obtain a works which holds the transformed cistrons in the works genome. Plant like sago that has slow tissue civilization and regeneration procedure therefore a system for efficient choice is of import. Poor choice system will let agents in the non-transformant cells to retroflex, particularly when the choice agents is less active after a in certain civilization incubation period and this will take to the production of chimeric. ( Parveez et al 1996 ) . Basta™ is used as a choice agent for M. sagu because it has been successfully used on the choice of the palm oil transformants. Moreovers the freshly planted sago thenar in the plantation showed high sensitiveness towards Basta ™ during everyday application to command the weeds which might be a good index for the presence of the new cistrons.
The efficiencies of the system were proved by DNA analysis. The transformants DNA were extracted and analyzed through Guam staining, Basta ™ choice media, PCR, point smudge analysis, RFLP-based DNA function and Southern Blot Analysis.
Therefore, the aims of this experiment are to:
develop the sago thenar suspension civilization system.
place the parametric quantities needed to genetically transformed sago thenar via Agrobacterium tumefaciens and/or atom barrage
screen for transformed cell lines and bring forth transgenic sago thenar
carry out molecular analysis and word picture of the transgene for stableness in transgenic sago thenar.