Evaluasi Galur Kedelai Transgenik Toleran Aluminium Pada Fasilitas Uji Terbatas

Some acid soil is potential for the agricultural development. Constraints for soybean production in the acid soils are Aluminum toxicity and macro nutrient deficiencies. Breeding for soybean varieties tolerant to acid soil is needed. This could be made through genetic engineering, by inserting acid tolerance genes into a soybean genome. Thirty one soybean lines (T0) had been obtained by insertion of Al tolerance genes (MaMt2) through an Agrobacterium mediated transformation, which nine of them contained MaMt2 gene based on PCR test. Further evaluation of those lines was carried out in the Biosafety Containment, where four T1 soybean lines were carrying MaMt2 gene, namely line GM2, GM5, GM10 and GM14. The study was aimed to evaluate the degree of tolerance of T2 generation of GM2, GM5, GM10 and GM14 lines to Al toxicity. Results showed that T2 line were able to grow in hygromicin media, indicating that those T2 lines were containing hygromicin resistant gene (hptII). Phenotypic analysis of T2 lines in four acid soil media treatments indicated that all lines could survive and grow on acid soil without liming and adding compost. GM2 line grew best on the acid medium than did other lines

Kultur Apeks Untuk Penyediaan Bibit Unggul Tebu Varietas PS864 Dan PS881

In vitro culturetechniques have become alternative to help overcome theproblems those are often encountered in the provision ofseeds through conventional means. Micropropagationthrough apex culture in sugarcane has several advantages,such as the produced plants have higher genetic stability,high multiplication rate, and more healthy seeds (especiallyvirus-free)., The aims of the the research were to produceseeds of two varieties of sugarcane, namely PS864 andPS881, through apex culture. Laboratory-scale research wasconducted at the Indonesian Center for AgriculturalBiotechnology and Genetic Resources Research andDevelopment (ICABIOGRAD), Bogor, while sowing seedsnursery was done in the Experimental Station of Gowa,South Sulawesi Assessment Institute for AgriculturalTechnology. The experiments consisted of initiation andregeneration of apexes, shoots multiplication, rootinginduction, and acclimatization of plantlets. Research resultsshowed the initiation and regeneration of PS864 and PS881through apex culture could be done on MS basic mediumcontaining 0.5 mg/l BAP. Shoot proliferation of both varietiesincreased in the second subculture. Addition of 1 mg/l BAPinto medium in the second subculture resulted in higheraverage number of shoots than that of 5 mg/l BAP, both forPS864 and PS881. Addition of 1 mg/l and 5 mg/l kinetinshowed no significant differences for shoot numberscompared to that of PS864 in medium containing 1 mg/lBAP. The average number of PS881 shoots in multiplicationmedia containing 5 mg/l kinetin was higher than that of 1mg/l kinetin. Increased concentrations of NAA and IBA from0.1 mg/l to 0.5 mg/l in the MS medium were correlated to theincreased number of roots in PS864 shoots. Meanwhile, onlyincreased concentration of NAA that affected rooting percentageof PS881. Acclimatization showed the percentage ofthe plantlets grown in polybags was higher than that directlygrown in planting bed. The primary seeds (G0) produced inthese experiments were ready to be reproduced again toobtain further stages

Regenerasi Kedelai Melalui Kultur Epikotil Dan Teknik Aklimatisasi

Regeneration of induced callus growth into whole plants is an important step in genetic transformation. Soybean is a recalcitrant plant regeneration process which can not always be repeated often (irreproducible). Therefore, the standard soybean callus regeneration method is difficult to obtain. This research was conducted to obtain a suitable medium for the regeneration and acclimatization techniques for soybean plant. The regeneration experiment used soybean sprouts of Sindoro cultivar on five kinds of media, with MS and B5 basal media plus a few types and concentrations of growth regulators, arranged on completely randomized design with three replications. Each replication consisted of five explants per bottle. Observations were made on the number of explants forming shoots, number of shoots of each explant, number of roots, and shoots length. Results showed that the culture of epicotyl had a high regeneration rate (>90%), and B5 media was the most suitable for plant regeneration of callus derived from epicotyl explants of soybean. Acclimatization of plantlet derived from epicotyl and one putative transgenic (TO) plant resulted in 99% degree of success. Plantlets from epicotyl culture of soybean Sindoro cultivar and putative transgenic plants were successfully grown in the greenhouse and subsequently produced pods

Penyisipan Gen Inhibitor α-Amilase Pada Plasmid Biner PCambia 1301

The experiment was conducted at the Molecular Biology Laboratory of the Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development, Bogor. The objective was to construct -ai gene on a binary plasmid pCambia 1301. This experiment was carried out using construction method by ligation process between fragments of α-ai gene from pTA3 plasmid and pCambia 1301 on HindIII site. The result of ligant transformation into E. coli DH5α was 182 surviving colonies on YEP medium containing kanamycin. DNA samples were obtained from 60 randomly selected colonies. The restriction pattern was tested by digesting each DNA sample using HindIII showed colonies containing two fragments expected of sizes wich are 11.837 and 4.887 kb. Two colonies are predicted containing of α-ai gene on its the binary plasmid. Advanced tests using restriction enzymes BamHI and XbaI showed two directions (right and left) of α-ai gene. The right direction was shown by pCambia-α-ai1 from colony number 43. This plasmid showed expected fragments of sizes 13.485 and 3.219 kb when digested with BamHI and two fragments of sizes 15.421 and 1.303 kb when digested with XbaI. The left direction was shown pCambia-α-ai2 from colony number 58. This plasmid also demon-strated expected fragments of sizes 15.026 and 1.698 kb when digested with BamHI and two fragments of sizes 13.082 and 3.642 kb when digested with XbaI. Both pCambia-α-ai1 and pCambia-α-ai2 were transformed into A. tumefaciens LBA4404

Analisis Fenotipik Progeni Tiga Galur Tomat Transgenik Partenokarpi Di Fasilitas Uji Terbatas (Phenotypic Analysis on Progenies of Three Transgenic Parthenocarpy Tomato Lines in Biosafety Containment)

Kebutuhan masyarakat akan buah tomat konsumsi dan untuk industri cenderung kian meningkat setiap tahunnya, sedangkan produksi tomat masih rendah. Hingga kini, produksi tomat nasional masih sangat rendah,yaitu 992.780 ton sehingga belum mencukupi kebutuhan pasar yang mencapai 1.230.000 ton. Beberapa upaya telah dilakukan pemerintah untuk meningkatkan produksi tomat, tetapi masih menemui beberapa masalah di lapangan. Perakitan varietas unggul tomat produktivitas tinggi dan tanpa biji (seedless) sangatlah diharapkan guna memenuhi kebutuhan masyarakat dan industri. Partenokarpi merupakan fenomena terjadinya pembentukan buah tanpa melalui proses penyerbukan dan atau pembuahan. Teknologi partenokarpi dapat digunakan untuk meningkatkan produksi tomat melalui peningkatan pembentukan buah (fruits setting) dan buah tanpa biji. Perakitan galur tomat partenokarpi melalui rekayasa genetik telah dilakukan di Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian (BB Biogen). Enam puluh galur (event) tomat transgenik T0 yang membawa gen DefH9-iaaM telah dihasilkan dan tiga galur di antaranya, yaitu OvR#14-4, OvM2#10-1, dan OvM2#6-2 telah terpilih sebagai galur terbaik berdasarkan karakter partenokarpinya. Galur tomat transgenik ini selanjutnya dievaluasi lebih lanjut secara molekuler dan fenotipik di rumah kaca dan rumah kasa fasilitas uji terbatas (FUT). Hasil analisis molekuler menunjukkan bahwa semua galur tomat transgenik yang diuji masih membawa gen DefH9-iaaM. Hasil evaluasi awal terhadap tiga galur tomat transgenik secara fenotipik juga menunjukkan sifat partenokarpi, yaitu meningkatnya jumlah buah, berat buah, dan berkurangnya jumlah biji (seedless). Analisis fenotipik lebih lanjut terhadap progeni ketiga galur tersebut pada percobaan ini menunjukkan adanya ekspresi fenotipik dari gen DefH9-iaaM, di mana galur OvR#14-4 memiliki ekspresi fenotipik partenokarpi lebih baik daripada galur OvM2#10-1 dan OvM2#6-2.KeywordsTomat; Rekayasa genetik; Gen partenokarpi; Galur transgenik; Analisis fenotipi

Transformasi Genetik Kedelai Dengan Gen Proteinase Inhibitor II Menggunakan Teknik Penembakan Partikel

An experiment was conducted at the Molecular Biology and Genetic Engineering Laboratory of BB-Biogen, Bogor with an objective to obtain transgenic soybean plants containing the proteinase inhibitor II (pinII) gene. The experiment consisted of three steps, i.e., optimalization of the soybean transformation technique using the gus gene; transformation of soybean using the pinII gene, and molecular analysis of the transformed soybean plants. Two type of explants (young embryo and cotyledon) were bombarded with pRQ6 plasmid containing the gus gene with the following treatment: Helium gas pressure (1100 psi and 1300 psi), shoot distance (5 and 7 cm), and number of bombardment (1x and 2x). The result of gus assay indicated that the best bombardment was done on young cotyledon explants with 1100 psi Helium pressure, shoot distance 5 cm, and 1x bombardment. Transformation of the soybean explant using the pinII gene (inside the pTWa plasmid) was conducted using the best bombardment treatment from the first activity. Two plants from c.v. Wilis (WP1, WP2) and three plants from c.v. Tidar (TP1, TP2, TP3) were recovered from regeneration and selection of the transformed explants. Molecular analysis of the regenerated plants using the PCR technique showed that only WP2 contained the pinII gene. This plant was fertile and will be used for further evaluation

Analisis Molekuler Gen Partenokarpi DefH9-RI-iaaM Pada Progeni Tomat Transgenik

The development of seedless tomato fruits will be more attractive to both consumers and industries. Seedless tomatoes can beproduced through parthenocarpy technology. Artificial parthenocarpy can be induced by conventional crossing, hormoneapplication, or genetic engineering. The development of parthenocarpic tomatoes through genetic engineering has been carriedout by inserting DefH9-iaaM parthenocarpic geneinto tomato genome via Agrobacterium tumefaciens mediated transformation.Sixty putative transgenic tomato lines were produced and three events (OvR1#14-4, OvM2#10-1, and OvM2#6-2) were selectedas the best events. The background of the tomato lines was Oval variety, and based on PCR results, the three selected linescontained DefH9-RI-iaaM in their genome. The objective of this research was to determine the integration of DefH9-RI-iaaMgene in the progenies of three transgenic tomatoes lines using PCR technique. The research was conducted in the laboratoryand Biosafety Containment Facility of Indonesian Center for Agricultural Biotechnology and Genetic Resources Research andDevelopment (ICABIOGRAD). Parental variety, Oval (neither transgenic nor in vitro cultured), and elite line of CL 6046 were usedas control plants. The results indicated that the progenies (T1, T2, and T3) of the three tomato lines contained the insert DefH9-RIiaaMgene

Genetic Mapping of SSR Markers in Eight Soybean Chromosomes Based on F2 Population B3462 X B3293

Genetic Mapping of SSR Markers in Eight SoybeanChromosomes Based on F2 Population B3462 x B3293. IMade Tasma, Ahmad Warsun, Dani Satyawan, SaptowoJ. Pardal, and Slamet. Aluminum toxicity is one of the maincontrains for cultivating soybean in acid soils. GeneticHak Cipta © 2011, BB-Biogenmapping of SSR markers is one step for detecting aluminumtoxicitytolerant QTLs in soybean. Another step is tophenotype the same population at various aluminum-toxicityenvironments. The objectives of this study were to analyzethe segregation of SSR markers in progenies of an F2population and map the markers in 8 soybean chromosomes.The F2 population was previously developed bycrossing the Al-tolerant parent B3462 and the Al-sensitiveparent B3293. Polymorphic SSR markers in the parents wereused to PCR amplify DNA of the 100 F2 progenies. PCRproducts were separated using agarose or polyacrylamidegels. A Chi-Square test was done with a null hypothesis thatprogenies segregated in a 1 : 2 : 1 ratio. Results showed that125 SSR markers were polymorphics in the parents. Out of125 polymorphic markers, 122 were segregated in theprogenies of the F2 population. Among the segregatingmarkers, 114 were segregated in a 1 : 2 : 1 ratio. Only 8markers (5.6%) did not follow the 1 : 2 : 1 ratio. One hundredand nineteen SSR markers were mapped in 8 soybeanchromosomes. These include 18 markers in chromosomeA2, 10 in B1, 16 (C1), 16 (F), 10 (G), 23 (J), 16 (L), and 10 (N).Total genetic maps covered was 1,194.8 cM with averagemap distances between two adjacent markers of 10.7 cM.Further SSR marker enrichment is required to fill in the gapsof several chromosomal regions. Genetic maps presented inthis study should be useful for detection of Al-toxicitytolerant QTLs in soybean