PERKEMBANGAN PEMANFAATAN, REGULASI, DAN METODE DETEKSI PRODUK REKAYASA GENETIKA PERTANIAN DI INDONESIA / Development of Utilization, Regulation, and Detection Methods of Agricultural Genetically Modified Products in Indonesia

Genetically modified crops (GM crops) have developed very fast globally, although to date controversies over the GM crop uses are still occurring. GM crops have been planted on over 191.7 million hectare area and cultivated in 26 countries in five continents. Biosafety of GM crops both globally and domestically are guaranteed through regulations made at the level of law, government regulations, related ministrial regulation including the guidelines. In general, those regulations have been implemented, thus the biosafety of GM crop utilization is guaranteed in Indonesia. Unfortunately, although Indonesia gave a certification for released permit for drought tolerant sugarcane, it only grown in a limited areas belongs to state-owned agricultural company (PTPN XI). The country has certified 27, 7, and 16 GM events for food, feed, and seeds for environment safety, respectively. The implementation of these regulations needs a monitoring system that is equipped with facilities of GMO detection laboratory with adequate capacity. Indonesia has several such laboratories. The methods of GMO detections have developed from very basic techniques, i.e. qualitative screening to the determination of specific events that define the type of trait of GMO, even quantitative detection, both single and multiplex. Each method has its own advantages. The capacities of GMO detection laboratory in Indonesia still need to be upgraded to master the fast-developing technology. The purpose of this review is to provide information on the development of global GM crops utilization including in Indonesia and the development of regulations and detection methods with their prospects and challenges.Keywords: Genetics, modification, regulation, detection methods AbstrakPemanfaatan tanaman produk rekayasa genetik (PRG) telah berkembang cepat dan mendunia walaupun sampai saat ini masih terjadi kontroversi. Luas penanamannya telah mencapai 191,7 juta ha dan ditanam oleh 26 negara di lima benua. Keamanan hayati PRG secara global maupun domestik telah dijamin oleh peraturan pada tingkat undang-undang, peraturan pemerintah, peraturan kementerian terkait, dan pedoman pelaksanaannya. Secara umum peraturan peraturan tersebut telah dijalankan sehingga keamanan hayati dari pemanfaatan PRG terjamin di Indonesia. Sayangnya di Indonesia PRG yang sudah diberi izin edar hanya ditanam secara terbatas seperti tebu toleran kekeringan di beberapa kebun milik PTPN. Indonesia juga telah memberikan sertifikat aman hayati pada beberapa varietas PRG diantaranya 27 PRG pangan, tujuh PRG pakan, dan 16 PRG benih (lingkungan). Implementasi peraturan yang telah ada memerlukan sistem pengawasan yang dilengkapi dengan fasilitas laboratorium deteksi PRG dengan kapasitas yang memadai. Indonesia telah mempunyai beberapa laboratorium tersebut. Metode deteksi PRG telah berkembang dari teknik yang sangat mendasar yaitu deteksi untuk skrining kualitatif PRG sampai teknik penentuan spesifik event yang menetapkan jenis/sifat PRG, bahkan teknik deteksi secara kuantitatif yang bersifat tunggal maupun multiplex. Metode-metode deteksi tersebut memiliki keunggulan masing-masing. Laboratorium penguji PRG di Indonesia masih perlu ditingkatkan kemampuannya dengan penguasaan teknologi yang berkembang dengan pesat. Makalah ini memberikan informasi perkembangan pemanfaatan PRG global termasuk di Indonesia dan perkembangan regulasi dan metode deteksi serta prospek dan tantangan.Kata kunci: Genetika, rekayasa, regulasi, metode deteks

Interaksi AtMEK1-EXGT Pada Arabidopsis Thaliana Pada Saat Terjadi Pelukaan

Proteininteractions occur within cellular level of stimulated plantcells to relay signals from receptors to production of response.AtMEK1-EXGT interaction had been detected in nontreatedArabidopsis. In this research, interaction betweenAtMEK1, a mitogen-activated protein kinase kinase ofArabidopsis thaliana, and EXGT, endoxyloglucan transferase,after the plant was wounded was examined usingco-immunoprecipitation and in vitro phosphorylation assay.The results demonstrated that EXGT interact with AtMEK1soon after and 10 minutes after wounding. In addition,AtMEK1 phosphorylation activity increased when increasedlevel of EXGT was incorporated into the reaction mixture.These indicate that EXGT amplifies wound-caused phosphorylationactivity of AtMEK1. The results elucidate part ofthe AtMEKK1-AtMEK1-AtMPK4 cascade which is stimulatedby wounding. How the complex interaction between EXGT,AtMEK1 and AtMPK4 fits within the cascade is remained tobe uncovered

Perancangan Dan Pembuatan Pemindah Barang Menggunakan Programmable Logic Controller

Banyak industri menghasilkan barang yang berbeda, baik dari segi ukuran maupun warna, namun hanya menggunakan sebuah konveyor secara bersama. Jadi barang-barang tersebut perlu dipisahkan atau dipindahkan menurut jenisnya. Selama ini proses pemindahan barang dilakukan dengan cara manual yaitu menggunakan tenaga manusia. Melalui tugas akhir ini akan dibuat pemindah barang berbentuk lengan robot yang relatif murah dan handal sebagai pengganti tenaga manusia. Pemindah Barang ini hanya akan memindahkan barang berupa kotak dengan ukuran10cm x10cm x10cm dengan warna hitam ke kiri atau putih ke kanan. Hal ini dimungkinkan karena alat ini mempunyai sensor ukuran dan sensor intensitas cahaya yang digunakan untuk mengenali warna kotak. Pemindah barang ini menggunakan tiga motor DC sebagai pengerak lengannya. Motor-motor ini digunakan agar lengan robot dapat melakukan gerakan mencengkram atau melepaskan kotak, gerakan naik ataupun turun dan gerakan memutar ke kiri atau ke kanan. Semua proses gerak pemindah barang ini dilakukan secara otomatis dengan kontrol dari PLC (Programmable Logic Controller). Pemindah barang yang dibuat ini telah melalui pengujian, baik dari sistem sensor ukurannya, sistem sensor intensitas cahaya, sistem lengannya maupun sistem secara keseluruhan dengan keberhasilan 100%. Jadi alat ini dapat diandalkan dalam penggunaannya

Interaksi AtMEK1-EXGT pada Arabidopsis thaliana pada Saat Terjadi Pelukaan

Proteininteractions occur within cellular level of stimulated plantcells to relay signals from receptors to production of response.AtMEK1-EXGT interaction had been detected in nontreatedArabidopsis. In this research, interaction betweenAtMEK1, a mitogen-activated protein kinase kinase ofArabidopsis thaliana, and EXGT, endoxyloglucan transferase,after the plant was wounded was examined usingco-immunoprecipitation and in vitro phosphorylation assay.The results demonstrated that EXGT interact with AtMEK1soon after and 10 minutes after wounding. In addition,AtMEK1 phosphorylation activity increased when increasedlevel of EXGT was incorporated into the reaction mixture.These indicate that EXGT amplifies wound-caused phosphorylationactivity of AtMEK1. The results elucidate part ofthe AtMEKK1-AtMEK1-AtMPK4 cascade which is stimulatedby wounding. How the complex interaction between EXGT,AtMEK1 and AtMPK4 fits within the cascade is remained tobe uncovered

MULTIFUNCTIONAL MUTANTS OF Azospirillum sp. WITH ENHANCED CAPABILITY OF SOLUBILIZING PHOSPHORUS, FIXING NITROGEN AND PRODUCING INDOLE ACETIC ACID

Azospirillum sp. have long been known as biofertilizer for plant growth because of its capability to produce phytohormones and fix nitrogen from the atmosphere. Multifunctional Azospirillum strain Aj Bandung 6.4.1.2 isolated in 2009 from cauliflower (Brassica oleracea) rhizosphere in Lembang, Bandung, West Java, was capable of fixing nitrogen, solubilizing tricalcium-phosphate, and producing phytohormone indole acetic acid (IAA). The study aimed to modify the multifunctions of Azos-pirillum sp. for better capability of fixing N2, solubilizing P, and producing IAA using ethyl methanesulfonate and 1-methyl-3-nitro-1-nitrosoguanidine (EMS) mutagen. The study was conducted at Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development (ICABIOGRAD) in 2010. The results showed that this strain was genetically mutagenized using EMS for better performance in solubilizing P, fixing N2 (nitrogenase activity), and producing phytohormone (IAA). The optimum concentration and the length of incubation time for the process have been determined. Nine selected mutants with increasing capability to solubilize P (determined by clear-zone formation on Pikovskaya’s medium) have been characterized for nitrogenase activities and IAA production compared to wild type Aj Bandung 6.4.1.2. The effect of mutagenesis on IAA produc-tion and nitrogenase activities varied among the mutans. Two mutants, AzM 3.7.1.16 and AzM 1.7.2.12, showed superiority in the production of IAA, while two mutants, AzM 1.5.1.14 and AzM 3.7.1.15, were superior in nitrogenase activities. The EMS mutagenesis of Azospirillum sp. showed enhanced dissolving capa-bility of unsoluble phosphate (tricalciumphosphate) and increased IAA production and nitrogenase activity.

Optimasi Sistem Regenerasi Dan Transformasi Padi Varietas Elit Indonesia

New ricevariety can be generated by means of transgenic approach.Transgenic rice researches have been conducted in manyinstitutions worldwide using Japonica, Indica, and Javanicavarieties. The most cultivated rice in Indonesia is Indica.Indica type is known to have low responsive tissues inculture and transformation media when compared toJaponica rice. This research activity is aimed to optimizeregeneration and transformation systems of the Indonesianelite rice varieties, so that good method can be achieved tobe used in the generation of transgenic elite rice varieties ofIndica type. The research consisted of two activities:regeneration and transformation optimizations in varieties ofDodokan (upland rice) and Inpari 6 (irrigated rice).Immature embryo was used as the explant in this research.The optimization studies used 2 types of media, NBH (N6salts and vitamins, cassamino acid 0.5 g/l, L-proline 0.5 g/l,sucrose 20 g/l, D-glucose 10 g/l, 2.4-D 2 mg/l, NAA 1 mg/l, BA1 mg/l, agarose Type I 5.5 g/l) and NBH-M (N6 macro salts,B5 micro salts, and vitamins, 0.3 g/l cassamino acid, 3 g/l Lproline,20 g/l sucrose, 3 mg/l 2.4-D, 1 mg/l NAA, 1 mg/l BAP,5.5 agarose Type I), 2 types of regeneration media, R1 (MSbase media and vitamis, 0.3 g/l glutamine, 30 g/l sucrose, 2mg/l kinetin, 1 mg/l NAA, 3 g/l phytagel) and R2 (MS basemedia and vitamins, 2 g/l cassamino acid, 20 g/l sucrose, 30mg/l sorbitol, 2.5 mg/l kinetin, 0.25 mg/l NAA, 3 g/l phytagel).Optimization transformation of Indonesian elite rice varietiesused developed an empty plasmid pCAMBIA 1301 containinghpt gene. The transformation was conducted using twotypes of co-cultivation media, K1 (N6 macro salts, B5 microsalts, and vitamins, 0.5 g/l cassamino acid, 0.5 g/l L-proline,20 g/l sucrose, 10 g/l glucose, 2 mg/l 2.4-D, 1 mg/l NAA, 1mg/l BAP, 0.1 mM acetosyringone) and K2 (N6 macro salts,B5 micro salts, and vitamins, 0.5 g/l cassamino acid, 0.5 g/l Lproline,20 g/l sucrose, 10 g/l glucose, 2 mg/l 2.4-D, 1 mg/lNAA, 1 mg/l BAP, 0.2 mM acetosyringone). The resultsshowed that Inpari 6 could form embryonic calli in NBHmedia and further regenerated well in R1 media (13.8%).The co-cultivation media K1 generated more selected calliwhich then generated green plant of young embryocompared to K2. Inpari 6 showed higher regeneration ratesafter transformation (3.6%) compared to Dodokan (0%).Molecular analysis showed that all 11 transformants (Inpari6) tested contained the hpt gene. These results are expectedto support the development of transgenic Indica ricegeneration in Indonesia

Pembentukan Populasi Mutan Azospirillum Dengan Menggunakan Transposon Untuk Sifat Superior Terhadap Pelarutan P

Azospirillum sp. which has the ability for nitrogenfixation and phosphate solubilization may support modernfarming in Indonesia that is mostly dependent on the USAgeof chemical fertilizer N, P, and K. Genetic quality ofAzospirillum was improved in this research to obtainsuperior characters toward phosphate solubilization so thatit can become more effective in use for farmers. To achievethis goal, Azospirillum was mutated by means ofelectroporation using transposon EZ-Tn5<kan-2>Tnp. Theelectrotransformation resulted in 20 out of 22 transformantstested contained the marker gen (npt). 10, 6 and 4 mutantshave increased, decreased and lost phosphate-solubilizingfunction, respectively. Mutant with elevated phosphatesolubilizingability may be selected further to be utilized asbiofertilizer while others may be useful for identification ofgenes responsible for phosphate solubilization

Optimasi Sistem Regenerasi dan Transformasi Padi Varietas Elit Indonesia

New ricevariety can be generated by means of transgenic approach.Transgenic rice researches have been conducted in manyinstitutions worldwide using Japonica, Indica, and Javanicavarieties. The most cultivated rice in Indonesia is Indica.Indica type is known to have low responsive tissues inculture and transformation media when compared toJaponica rice. This research activity is aimed to optimizeregeneration and transformation systems of the Indonesianelite rice varieties, so that good method can be achieved tobe used in the generation of transgenic elite rice varieties ofIndica type. The research consisted of two activities:regeneration and transformation optimizations in varieties ofDodokan (upland rice) and Inpari 6 (irrigated rice).Immature embryo was used as the explant in this research.The optimization studies used 2 types of media, NBH (N6salts and vitamins, cassamino acid 0.5 g/l, L-proline 0.5 g/l,sucrose 20 g/l, D-glucose 10 g/l, 2.4-D 2 mg/l, NAA 1 mg/l, BA1 mg/l, agarose Type I 5.5 g/l) and NBH-M (N6 macro salts,B5 micro salts, and vitamins, 0.3 g/l cassamino acid, 3 g/l Lproline,20 g/l sucrose, 3 mg/l 2.4-D, 1 mg/l NAA, 1 mg/l BAP,5.5 agarose Type I), 2 types of regeneration media, R1 (MSbase media and vitamis, 0.3 g/l glutamine, 30 g/l sucrose, 2mg/l kinetin, 1 mg/l NAA, 3 g/l phytagel) and R2 (MS basemedia and vitamins, 2 g/l cassamino acid, 20 g/l sucrose, 30mg/l sorbitol, 2.5 mg/l kinetin, 0.25 mg/l NAA, 3 g/l phytagel).Optimization transformation of Indonesian elite rice varietiesused developed an empty plasmid pCAMBIA 1301 containinghpt gene. The transformation was conducted using twotypes of co-cultivation media, K1 (N6 macro salts, B5 microsalts, and vitamins, 0.5 g/l cassamino acid, 0.5 g/l L-proline,20 g/l sucrose, 10 g/l glucose, 2 mg/l 2.4-D, 1 mg/l NAA, 1mg/l BAP, 0.1 mM acetosyringone) and K2 (N6 macro salts,B5 micro salts, and vitamins, 0.5 g/l cassamino acid, 0.5 g/l Lproline,20 g/l sucrose, 10 g/l glucose, 2 mg/l 2.4-D, 1 mg/lNAA, 1 mg/l BAP, 0.2 mM acetosyringone). The resultsshowed that Inpari 6 could form embryonic calli in NBHmedia and further regenerated well in R1 media (13.8%).The co-cultivation media K1 generated more selected calliwhich then generated green plant of young embryocompared to K2. Inpari 6 showed higher regeneration ratesafter transformation (3.6%) compared to Dodokan (0%).Molecular analysis showed that all 11 transformants (Inpari6) tested contained the hpt gene. These results are expectedto support the development of transgenic Indica ricegeneration in Indonesia