Judul: Morphological and molecular analysis of jatropha curcas l.related to aluminum stress as a preliminary study to develop transgenic plant
Abstrak: As a response to a progressive price increase of fossil based oil in the world, various initiatives to develop a reliable source of renewable energy become a major global attention. One potentially promising option is biofuel since these are derived from biomass. Biofuel crops should be planted on land that is considered marginal. A crop that is often cited as ideal for growing on marginal land including acid soil with high Al solubility, is the physic nut, Jatropha curcas. However, the major problem of the plant cultivation in acid soil is the decreasing productivity. Jatropha plants are not, by nature, tolerant to Al stress. A promising approach to overcome this limitation is to develop a transgenic Jatropha plant that can withstand to severe Alstress. Thus it is possible to use TaALMT (Triticum aestivum Al-activated malate transporter 1) gene which encodes the member of a novel membrane protein family that functions as an anion channel to mediate malate efflux from plant roots cells in the presence of extracellular Al3+. Investigating the physiology and molecular basis of Al-stress tolerance during seedling development is an essential prerequisite before developing Al-tolerant Jatropha plant by genetic engineering. Among six populations of Jatropha curcas L., IP-2P population could be categorized as slightly tolerant to Al stress, but it had uniform seed viability, and high productivity character which became the reason to use it as a plant material to develop transgenic Jatropha plant. For preliminary study, J. curcas IP-2P was choosen to be used for gene expression analysis by Quantitative Real-Time Polymerase Chain Reaction (qRT PCR). To normalize mRNA expression in the gene expression analysis we used actin gene. Because there is no information of actin gene nucleotide sequence from J. curcas we isolated Jatropha actin (JcACT) by PCR. Three cDNA of actin genes from J. curcas L. IP-2P had been isolated. The analysis revealed that Al stress induced ALMT transcript accumulation both in roots and leaves of non transgenic J. curcas IP-2P. Low pH treatment alone could also induce transcript accumulation both in the root apices and leaf but at lower rates than that observed for Al. This result indicated that response of Jatropha seedlings were not specific to Al stress but also low-pH stress. The TaALMT gene was successfully introduced into Jatropha curcas L. IP-2P via Agrobacterium tumefaciens-mediated transformation. Two putative transformant shoots were regenerated following selection on the bispyribac-containing medium. The presence of the transgene in the genome of transgenic plants was confirmed by PCR. The results indicated that the exogenous TaALMT1 gene was successfully integrated into the genome of J. curcas IP-2P plants.
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Judul: Transformasi Genetik Tanaman Jarak Pagar (Jatropha curcas L.) dengan Gen Metallothionein Tipe 2 dari Melastoma malabathricum melalui Perantara Agrobacterium tumefaciens
Abstrak: The main reason why we use marginal land for non-food crops like jatropha is the limited potential land to cultivate it. The condition of marginal land with low fertility, low pH and high content of heavy-metal can caused inhibit the growth and decreasing production of crop. The tolerance of plants to metal toxicity can be genetically improved by introducing and expressing a gene associated with the metal tolerance. One of these genes is MaMt2, a gene coding type 2 metallothionein that isolated from Melastoma affine. Metallothionein is a cystein-rich protein so it can binding various types of metal. The objective of this research was to obtain increase resistance of jatropha to heavy-metal by integrated MaMt2 gene to jatropha and molecular analysis from transgenic plant. Genetic transformation was performed by using Agrobacterium tumefaciens strain LBA4404 which containing pIG6-SMt2 plasmid. pIG6-SMt2 plasmid contains of MaMt2 gene, hptII gene (hygromycin phosphotransferase) under ubiquitin promoter and NOS (nopaline synthase) terminator. Cotyledon from 2 weeks-old of jatropha cultivar IP-2P was used. The non-transformed explants regenerated 14 days in the non selective regeneration medium while the transformed ones regenerated 18 days in the selective regeneration medium. The average of regenerated shoot each explant was 2.1 for non-transformed explants and 1.6 for transformed ones. Putative transgenic shoots were selected in two step by using medium containing 1.5 mg l-1 and 2.5 mg l-1 of hygromycin successively. This research resulted 25 putative transgenic shoots. Molecular analysis was performed to determine the integration of MaMt2 gene under control for ubiquitin promoter in transgenic shoots by PCR. PCR was carried out by using with UbiQF-NosTR and SMt2F-NosTR combination primers. PCR of transgenic shoots resulted 1160 bp and 526 bp fragments respectively. Molecular analysis showed that three of 15 putative transgenic shoots are transgenic shoots containing MaMt2 gene under control of ubiquitin promoter.
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Judul: Penyebaran beberapa jenis cetacean subordo odontoceti di perairan laut Sawu bagian Timur, Nusa Tenggara Timur
Abstrak: Cetacean yang ditemukan di perairan Laut Sewu pada Tanggal 27-28 Desember 2005 terdiri atas 8 spesies dan 1669 ind dengan total 76 sighting. Panjang total transek adalah 400,22 km meliputi 400,22 km daerah pengamatan.
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