Edy Listanto
Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development

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EFFECTS OF LATE BLIGHT RESISTANT POTATO CONTAINING RB GENE ON THE SOIL MICROBES, PESTS AND PLANT DISEASES Eny Ida Riyanti; Edy Listanto; Alberta Dinar Ambarwati
Indonesian Journal of Agricultural Science Vol 15, No 2 (2014): October 2014
Publisher : Indonesian Agency for Agricultural Research and Development

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/ijas.v15n2.2014.p47-54

Abstract

Late blight caused by Phytophthora infestans is an important disease on potato.  Several potato hybrids have been generated by crossing local varieties (Atlantic and Granola) with Katahdin SP951 which contains late blight resistance gene RB.  Prior to release, these hybrids need to be evaluated for their environ-mental effects on non-target organisms and natural pests and diseases. The objectives of the study were to investigate the effect of LBR potato hybrids on beneficial soil microbes, pests and diseases. The trial was conducted in the confined field trial (CFT) in Lembang, West Java. The parental non-transgenic (NT) clones (Granola, Atlantic and Katahdin) and LBR hybrids (four clones of Atlantic x Katahdin SP951 hybrids; 10 clones of Granola x Katahdin SP951) were planted at a plant spacing of 30 cm x 70 cm. Fungicide applications were used as treat-ments (no spray, five and twenty times sprays). The experi-ment was arranged in a randomized completely block design with three replications. The parameters determined were popula-tions of N2 fixing and P solubilizing bacteria, soil C/N ratio as well as natural pests and diseases. The results showed that the transgenic LBR potato hybrids did not have negative effect on N fixing bacteria. The bacterial populations were around 1010-11 cells g-1 soil before planting, 1012 cells at 1.5 months after planting (MAP) and 108 cells after harvest. For P- solubilizing bacteria, their populations were 1010 cells before planting, 1012 cells at 1.5 MAP and 1011 cells g-1  soil after harvest. The soil C/N ratio of the transgenic plot was not statistically different compared to non-transgenic plot, i.e. 12-15 before planting, 10-11 at 1.5 MAP, and 10 after harvest in non-spray plot. Pests and diseases such as Alternaria solani, Liriomyza, potato tubber moth, aphid and mites on the transgenic and non-transgenic plots were statistically not different. The resistance score for A. solani was 7.2 (parental tansgenic) and 7.6 (parental non-transgenic); for Liriomyza it was 2.07 (parental transgenic) and 2.32 insect per plant (parental non-transgenic), the PTM was 0.63 (parental transgenic) and 0.73 insect per plant (parental non-transgenic), aphid and mites were 0.75 (parental transgenic) and 1.68 insects per plant (parental non-transgenic). The study indicated that LBR potato hybrids did not have any negative impacts on non-target organisms.
Agrobacterium tumefaciens-MEDIATED IN-PLANTA TRANSFORMATION OF INDONESIAN MAIZE USING pIG121Hm-Cs PLASMID CONTAINING nptII AND hpt GENES Edy Listanto; Eny Ida Riyanti; Sustiprijatno Sustiprijatno
Indonesian Journal of Agricultural Science Vol 17, No 2 (2016): October 2016
Publisher : Indonesian Agency for Agricultural Research and Development

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/ijas.v17n2.2016.p49-56

Abstract

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.