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All Journal Indonesian Journal of Agricultural Science Jurnal Hortikultura JURNAL BIOLOGI INDONESIA Makara Journal of Science
Alberta Dinar Ambarwati
Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, Jl. Tentara Pelajar 3A, Bogor 16111
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology

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RESISTANCE EVALUATION ON POPULATIONS OF CROSSES BETWEEN TRANSGENIC POTATO KATAHDIN RB AND NON-TRANSGENIC ATLANTIC AND GRANOLA TO LATE BLIGHT (Phytophthora infestans) IN CONFINED FIELD TRIAL Ambarwati, Alberta Dinar; Herman, Muhammad; Purwito, Agus; Sumaraw, Sientje Mandang; Aswidinnoor, Hajrial
Indonesian Journal of Agricultural Science Vol 12, No 1 (2011): April 2011
Publisher : Indonesian Agency for Agricultural Research and Development - MOA

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Abstract

Late blight resistance gene (RB gene) isolated from Solanum bulbocastanum, is a broad resistance gene against all races of Phytophthora infestans. The gene was transformed into Katah-din event SP904 and SP951 using Agrobacterium tumefaciens and these transgenic plants have been crossed with susceptible potato cultivars Atlantic and Granola. Populations of the crosses have been molecularly characterized for the integration of the RB transgene. The study aimed to evaluate the resistance of the populations of crosses between transgenic Katahdin RB  and susceptible non-transgenic parents (Atlantic and Granola) to late blight in a confined field trial at Pasir Sarongge, Cianjur, West Java. A total of 84 clones originated from four popula-tions were evaluated for resistance to late blight. These included 22 clones of Atlantic x transgenic Katahdin SP904, 16 clones of Atlantic x transgenic Katahdin SP951, 19 clones of Granola x transgenic Katahdin SP904, and 27 clones of Granola x transgenic Katahdin SP951. Observations of the late blight infection were conducted when late blight symptoms were detected, i.e. at 56, 60, 63, 70, and 77 days after planting (DAP). The result showed there were high variations in the resistance level of all the 84 clones tested. Clones of crosses between susceptible parents (Atlantic or Granola) and resistant parents (transgenic Katahdin SP904 or Katahdin SP951) showed a similar pattern based on the area under disease progress curve (AUDPC) value, i.e. 377.2 greater than the AUDPC of the resistant parents (180.1), but smaller than that of the susceptible parents (670.7). Observation at 77 DAP resulted four resistant potato clones having resistance score of 7.0-7.6, higher than the transgenic parents Katahdin SP904 (4.6) and Katahdin SP951 (6.8), i.e. clone B8 (Atlantic x transgenic Katahdin SP951) with resistance score of 7.6 and clones B26 (Atlantic x transgenic Katahdin SP951), C183 (Granola x transgenic Katahdin SP904), and D89 (Granola x transgenic Katahdin SP951) with resistance score of 7. These four transgenic potato resistant clones need to be further developed as promising potato clones to late blight.
Pengujian Ketahanan Klon-klon Hasil Silangan Tanaman Kentang Transgenik dengan Nontransgenik terhadap Penyakit Hawar Daun Phytophthora infestans di Lapangan Uji Terbatas Ambarwati, Alberta Dinar; Herman, Muhamad; Lisanto, Edi; Suryaningsih, Euis; Sofiari, Eri
Jurnal Hortikultura Vol 22, No 2 (2012): Juni 2012
Publisher : Indonesian Center for Horticultural Research and Development

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Abstract

STRAK. Tanaman kentang transgenik Katahdin event SP904 dan SP951 mengandung gen RB, yang diisolasi dari spesies liar kentang diploid Solanum bulbocastanum. Gen RB mempunyai ketahanan yang bersifat  durable dengan spektrum yang luas terhadap ras-ras Phytophthora  infestans di Amerika Serikat. Dalam perakitan tanaman kentang tahan penyakit hawar daun P. infestans di Indonesia, transgenik Katahdin dijadikan sebagai donor tahan dalam persilangan dengan varietas rentan Atlantik dan Granola. Klon-klon hasil silangan dianalisis secara molekuler mengandung gen RB. Penelitian dilakukan untuk menguji ketahanan klon-klon hasil silangan tanaman kentang transgenik dengan nontransgenik terhadap isolat P. infestans di lapangan uji terbatas (LUT) yang berlokasi di Balai Penelitian Tanaman Sayuran (Balitsa), Lembang dari bulan Oktober 2009 sampai dengan Maret 2010. Klon-klon yang diuji ialah 12 klon hasil silangan Atlantik x transgenik Katahdin SP904 (A); 15 klon hasil silangan Atlantik x transgenik Katahdin SP951 (B); 17 klon hasil silangan Granola x transgenik Katahdin SP904 (C); dan 20 klon hasil silangan Granola x transgenik Katahdin SP951 (D). Atlantik dan Granola digunakan sebagai kontrol rentan, sedangkan transgenik Katahdin sebagai kontrol tahan. Pengamatan dimulai ketika muncul gejala awal, yaitu pada 26, 32, 39, 46, dan 53 hari setelah tanam. Ketahanan tanaman semakin menurun dengan bertambahnya periode pengamatan, diikuti meningkatnya intensitas penyakit dan AUDPC. Semua klon yang diuji menunjukkan keragaman dalam ketahanan fenotipik terhadap hawar daun P. infestans. Klon-klon hasil silangan Atlantik x transgenik Katahdin SP951 mempunyai nilai AUDPC 697, yang hampir sama dengan transgenik Katahdin SP904 yaitu 698,5. Klon-klon Granola x transgenik Katahdin SP951 mempunyai nilai AUDPC  687,5 lebih kecil dibandingkan transgenik Katahdin SP904. Hasil penelitian ini juga menunjukkan bahwa klon-klon tersebut mempunyai ketahanan yang lebih tinggi dibandingkan transgenik Katahdin SP904. Pada pengamatan 46 hari setelah tanam atau 20 hari setelah infeksi diperoleh tiga klon tahan yaitu B49 (skor 7,5), C111 (skor 7,1), dan D26 (skor 7,3). Ketahanan ini lebih tinggi daripada transgenik Katahdin SP904 (skor 5,1) dan transgenik Katahdin SP951 (skor 6,4). ABSTRACT. Ambarwati, AD, Herman, M, Listanto, E, Suryaningsih, E and Sofiari, E 2012. Resistance Testing on Transgenic and Nontransgenic Potato Clones Against Late Blight Phytophthora  infestans in Confined Field Trial.  Transgenic potato Katahdin event SP904 and  SP951 containing RB gene, which were isolated from a wild diploid potato species, Solanum bulbocastanum. RB gene showed durable resistance with broad spectrum to all known races of  P. infestans in the USA. In development of  potato resistant to late blight P. infestans in Indonesia, Katahdin transgenic were used as a resistant donor and crossed with susceptible varieties i.e. Atlantic and Granola. Clones derived from the crossing were molecularly analyzed and had RB gene contain. Experiment was conducted to assess the resistance of the clones derived from crossing of Katahdin transgenic and nontransgenic to P. infestans in confined field trial (CFT), located at the Indonesian Vegetable Research Institute (IVEGRI), Lembang from October 2009 to March 2010. Several clones tested were 12 clones of Atlantic x Katahdin transgenic SP904 (A); 15 clones of Atlantic x Katahdin transgenic SP951 (B); 17 clones of Granola x Katahdin transgenic SP904 (C); and 20 clones of Granola x Katahdin transgenic SP951 (D). Atlantic and Granola were used as susceptible control whereas Katahdin transgenic as resistant control. Observation was started as late blight symptoms and detected at 26, 32, 39, 46, and 53 days after planting. Plant resistance decreases with increasing period of observation, followed by increasing disease intensity and AUDPC. All clones tested showed variation in phenotypic resistance to late blight P. infestans. Clones derived from crossing of Atlantic x Katahdin transgenic SP951 had AUDPC score 697 and almost similar to Katahdin transgenic SP904 (698.5). Clones derived from crossing of Granola x Katahdin transgenic SP951 had AUDPC score 687.5 and smaller than Katahdin transgenic SP904. The results also indicated that these clones had higher resistance than Katahdin transgenic SP904. Observation at 46 days after planting or 20 days after infection resulted three resistant clones i.e. B49 (score 7.5), C111 (score 7.1); and D26 (score 7.3).  This resistance was higher than Katahdin transgenic SP904 (score 5.1) and Katahdin transgenic SP951 (score 6.4).
EFFECTS OF LATE BLIGHT RESISTANT POTATO CONTAINING RB GENE ON THE SOIL MICROBES, PESTS AND PLANT DISEASES Riyanti, Eny Ida; Listanto, Edy; Ambarwati, Alberta Dinar
Indonesian Journal of Agricultural Science Vol 15, No 2 (2014): October 2014
Publisher : Indonesian Agency for Agricultural Research and Development - MOA

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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.
Klon-klon Kentang Transgenik Hasil Persilangan Terseleksi Tahan terhadap Penyakit Hawar Daun Phytophthora infestans Tanpa Penyemprotan Fungisida di Empat Lapangan Uji Terbatas Ambarwati, Alberta Dinar; ,, Kusmana; Listanto, Edy
JURNAL BIOLOGI INDONESIA Vol 11, No 2 (2015): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v11i2.2191

Abstract

ABSTRACTThe 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. Keywords : transgenic potato, hybrid clones, Phytophthora infestans, confined field trial 
KLON-KLON KENTANG TRANSGENIK HASIL PERSILANGAN TERSELEKSI TAHAN TERHADAP PENYAKIT HAWAR DAUN PHYTOPHTHORA INFESTANS TANPA PENYEMPROTAN FUNGISIDA DI EMPAT LAPANGAN UJI TERBATAS Ambarwati, Alberta Dinar; ,, Kusmana; Listanto, Edy
JURNAL BIOLOGI INDONESIA Vol 11, No 2 (2015): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v11i2.2191

Abstract

ABSTRACTThe 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. Keywords : transgenic potato, hybrid clones, Phytophthora infestans, confined field trial 
Bioinformatics Toxicity Assessment of RB Protein from Transgenic Potato Resistant to Phytophthora infestans Hadiarto, Toto; Ambarwati, Alberta Dinar
Makara Journal of Science Vol. 23, No. 1
Publisher : UI Scholars Hub

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Abstract

The genetically engineered potato cultivar Katahdin SP951 is resistant to late blight disease, which is caused by Phytophthora infestans. The biosafety and food safety of this cultivar should be assessed prior to its commercialization. Toxicity is one of the parameters tested in the food safety evaluation of transgenics. Toxicity testing includes the bioinformatics analysis of the homology of the RB protein of Katahdin SP951 with known proteins. Therefore, this study aimed to perform the bioinformatics analysis of Katahdin SP951 RB protein against homologous toxin proteins. Bioinformatics analysis was conducted by first translating the RB gene into an amino acid sequence by using the Emboss Transeq program from the EMBL-EBI website. The Shuffle Protein Program was then applied to obtain the randomized amino acid sequences of the RB protein. The Basic Local Alignment Search Tool (BLAST) Protein search program was utilized to identify proteins with high similarity and homology. Moreover, BLAST Suite-2 was used to analyze the homology between two or more sequence alignments. Results showed high homology between the sequences of the RB protein and those of known resistance (R) proteins with an E value of less than 0.22. This result may be attributed to the presence of numerous R proteins in plants. Further analysis indicated that the sequence of the RB protein had extremely nonsignificant homology with sequences of proteins known to be toxic in the Entrez protein database of National Center for Biotechnology Information. Therefore, the RB protein is nontoxic.
Co-Authors Agus Purwito Edi Lisanto Edy Listanto Eny Ida Riyanti Eri Sofiari Euis Suryaningsih HAJRIAL ASWIDINNOOR Kusmana ,, Kusmana Muhamad Herman MUHAMMAD HERMAN SIENTJE MANDANG SUMARAW Toto Hadiarto