Pertumbuhan Padi Varietas Ciherang Setelah Diinokulasi dengan Azospirillum Mutan Multifungsi

Modern agriculture is very closely related to the application of fertilizer to induce plants grow. The application of bio-fertilizers is expected to reduce the negative impact of chemical fertilizers. The purpose of this study was to determine the effect of multi-functional Azospirillum N2 fixation, P solubility and IAA production on the growth of Ciherang rice in pot experiment in greenhouse BB Biogen. The experiment treatment were 3 types of inoculation (non-inoculation, inoculation using wildtype Aj Bandung 6.4.1.2 and the mutant isolate of AJM 3.7.1.14 ), and 4 levels of fertilizer application (non-fertilization, a quarter dose, a half dose, and the real dose of fertilization on rice in lowland). The Azospirillum isolates were used wildtype isolate Aj Bandung 6.4.1.2 and mutant isolate AJM 3.7.1.14 that was isolated and mutated using ethyl methanesulfonate (EMS) in BB Biogen. Seeds of Ciherang rice were inoculated using Azospirillum at cell density 106 cell/ml in different seedling tray. After 14 days, the seedlings were transferred to planting pots which consist of 3 plants per pot. Parameters observed were plant height, number of tillers, number of panicles per hill, wet and dry weight of panicles per hill, weight of 100 seeds, N and P content of the stover. The results showed that both wild-Azospirillum and mutant inoculum had no effect on the vegetative growth of Ciherang, but showed significant effect on the number of panicle per hill, grain weight per hill and dry weight of seeds per panicle. The use of Azospirillum and N fertilizer combination affected the growth and rice yields, also reduced chemical fertilizer application

Uji Ketahanan Galur-galur Kentang Transgenik Hasil Transformasi Dengan Gen Rb Terhadap Penyakit Hawar Daun (Phytophthora Infestans) Di Kp Pasirsarongge, Cianjur

Resistance test strains of transgenic potatoes transformed with RB gene to late blight (Phytophthora infestan) in KP Pasirsarongge, Cianjur. Potato late blight caused by Phytophthora infestans (P. infestans) (Mönt.) de Barry continues to be one of the most important crop diseases of all time. Genetic engineering of potato using RB gene for resistant plant to this disease is the most effective and environmental friendly to prevent widespread of late blight. This research aims to perform resistance of transgenic potato lines containing RB gene to lateblight (P. infestans) in Pasirsarongge, Cianjur field trial station. The first generation of transgenic lines were planted on polybag containing soil:manures using randomized complete block design. Tested plant inoculation was done naturaly from inoculum source from border row (Granola) that has been planted at one month before. The symptom was observed at one month after planting and damage scoring was done every three days for five times. Twenty two transgenic lines of tested plant showed various resistance respond to late blight (P. infestans) attack. Three transgenic lines showed highly resistance to late blight (P. infestans) were lines 11, 24, and 25, one transgenic line has resistant level was line 6

Penentuan Alergenisitas Protein Gen RB Pada Kentang Produk Rekayasa Genetika Berdasarkan Studi Bioinformatika

Genetically modified products (GMP) of Katahdin potato event SP951 containing RB gene resistant to late blight diseasescaused by Phytophtora infestans has been developed in the USA. This Katahdin SP951 potato has been crossed with localvarieties Atlantic and Granola for its development in Indonesia. In the release process, the GMP potato should be tested forenvironmental and food safety. One of the food safety assessment needs to be done by determining allergenicity of RB proteinwhether it is potential as allergen. This research aims to translate the RB gene sequence into RB protein sequence andinvestigate the potential RB protein as an allergen through bioinformatic studies. This study was performed based on thealignment with available protein allergens from available database websites. The predicted RB protein obtained from 2,913amino acids RB gene was a 971 amino acids length protein with ATG as a start codon and TAA as a stop codon. Bioinformaticsstudies of RB protein were performed using www.allergenonline.com, consisted of three searches, i.e. full-length search byFASTA, 80 amino acids search by FASTA, and 8 amino acid exact matches. For full-length alignment search, there are threeallergen proteins similar with RB protein sequence with the percentage identity of <35%, while for alignment with 80 aminoacids and 8 amino acids did not show similarity with any allergen protein in the database. It can be concluded that RB proteindid not have any potential as an allergen, as according to Codex Alimentarius guidelines for full-length alignment search, onlyprotein with identity greater than >50% indicating possible cross reactivity with protein allergen

Agrobacterium Tumefaciens-mediated In-planta Transformation of Indonesian Maize Using Pig121hm-cs Plasmid Containing Nptii and Hpt Genes

Maize (Zea mays L.) productivity in Indonesia is challenged to be increased using genetic engineering. Recent advances in Agrobacterium tumefaciens-mediated in-planta transforma-tion makes it possible to transform maize with low cost, and simple method. This study aimed to confirm pIG121Hm-Cs plasmid in A. tumefaciens, and to estimate the efficiency level of A. tumefaciens-mediated in-planta transformation of Indonesian maize by using pIG121Hm-Cs plasmid containing nptII and hpt genes. A series of studies were conducted including confirmation of gene construct of pIG121Hm-Cs plasmid in A. tumefaciens, transformation of four maize lines through A. tumefaciens-mediated in-planta technique, acclimatization of transformant plants and molecular analysis of selected plants using polymerase chain reaction (PCR). The pIG121Hm-Cs plasmid was confirmed via PCR analysis using specific primers of nptII and hpt genes and resulted 700 bp and 500 bp for fragments of nptII and hpt, respectively. After selection, acclimatization and molecular analysis steps, the efficiency levels of transformation of four maize lines were low, ranging from 3.8% to 12.8%. The level of transformation efficiency of ST-27 line was the highest accounting for 12.8% of 45 planted embryos on selection medium based on PCR analysis using specific primer for nptII gene. Overall, A. tumefaciens-mediated in planta transformation on maize floral pistil in this study proved to be successful and rapid. Therefore, this enhanced transformation method will be beneficial for Indonesian maize genetic engineering

Klon-klon Kentang Transgenik Hasil Persilangan Terseleksi Tahan Terhadap Penyakit Hawar Daun Phytophthora Infestans Tanpa Penyemprotan Fungisida Di Empat Lapangan Uji Terbatas

The use of resistant varieties is an appropriate alternative in controlling the late blight, a major diseases on potato, caused by the fungus Phytophthora infestans. The development of late blight resistant potato was done through hybridization between non-transgenic Atlantic or Granola with RB transgenic Katahdin SP904 and SP951. The hybrid clones which have been positively contained the RB gene were evaluated for the resistance to P. infestans in four Confined Field Trials (CFTs) i.e. Pasir Sarongge (2008), Lembang (2009-2010), Pangalengan (2010-2011) and Banjarnegara (2011-2012). There are twelve selected hybrid clones which were resistant to P. infestans both in each location of CFT or in four locations were obtained. These clones consist of five clones from crosses of Atlantic and trangenic Katahdin SP951 (B35, B169, B163, B11, B162) and seven clones from crosses of Granola and transgenic Katahdin SP951 (D76, D12, D25, D48, D38, D37, D15). The selected hybrid clones showed resistance to P. infestans until 14 to 18 days after infection or about 40 to 45 days after planting, in the absence of fungicide spraying. The hybrid clones had a resistance score varied from 7,65 to 8,23 and were significantly different from the parents Atlantic and Granola, with a resistance score of 3,6 and 3,45, respectively. This was also supported by AUDPC values, which showed that AUDPC of the hybrid clones were in the range between Atlantic or Granola and transgewnic Katahdin SP951. This indicate that the resistance level of the hybrid clones is in the range between susceptible and resistant check. The resistant hybrid clones are valuable genetic resources for late blight resistance breeding programs, particularly in reducing the frequency of fungicide applications

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

Induksi Dan Regenerasi Kalus Jagung Yang Ditransformasi Dengan Gen CsNitr1-L Melalui Penembakan Partikel

The success in development of transgenic plants is influenced by the regeneration system. The objective of the study was toassess the response of maize genotypes to regeneration system of organogenesis and embryogenesis, after transformed withCsNitr1-L gene through particle bombardment. Induction and callus regeneration of maize immature embryos of inbred linesUlt:cm.1#, ARC 178-123-112-XB3, and AZ2 were conducted through organogenesis, whereas those inbred lines AZ1, AZ2,P4G19(S)C2.59.3.3.1.3 and P4S3.29.4.4.1 were conducted through embryogenesis somatic. Transformation of CsNitr1-L gene wasdone with the distance of bombardment of 7 cm and 9 cm and calli were then selected using 10 mg/l hygromycin. All explants(100%) of inbred lines Ult:cm.1# formed organogenic callus, while callus formation of ARC 178-123-112-XB3 was 94.3% and AZ2was 60.5%. Ult:cm.1# was the most responsive line to the regeneration of organogenesis and produced 24 green shoots,compared with ARC 178-123-112-XB3 which produced one green shoot and AZ2 that did not produce green shoots. The highestpercentage of embryogenic calli formed through somatic embryogenesis was obtained on inbred lines AZ1 (85.4%) and thelowest was on P4S3.29.4.4.1 (18.9%). Inbred lines AZ1 had the highest percentage of regeneration (50.7%) and produced 62plants, followed by P4G19(S)C2.59.3.3.1.3 that produced 17 plants (2.8%) and P4S3.29.4.4.1 which produced two plants.Preliminary identification on 31 putative transgenic plants through PCR analysis produced 22 plants (70.96%) that contained nptIIgene