THE SECOND INTERNATIONAL CONFERENCE ON GENETIC RESOURCES AND BIOTECHNOLOGY: Harnessing Technology for Conservation and Sustainable Use of Genetic Resources for Food and Agriculture
THE SECOND INTERNATIONAL CONFERENCE ON GENETIC RESOURCES AND BIOTECHNOLOGY: Harnessing Technology for Conservation and Sustainable Use of Genetic Resources for Food and Agriculture
Keragaman Jumlah Salinan Transgen CsNitr1-L pada Galur Transforman T0 Padi Kultivar Nipponbare. P... more Keragaman Jumlah Salinan Transgen CsNitr1-L pada Galur Transforman T0 Padi Kultivar Nipponbare. Perakitan tanaman transgenik dengan menggunakan bantuan Agrobacterium tumefaciens menghasilkan penyisipan transgen yang berbeda, baik dalam jumlah salinan maupun letak transgen dalam genom tanaman. Penelitian ini bertujuan untuk menganalisis keberadaan kimera dan jumlah salinan pada tiap anakan dalam satu rumpun dan beberapa rumpun padi Nipponbare transforman T0 yang berasal dari kalus yang sama. Gen CsNitr1-L pada plasmid biner pCAMBIA1300 ditranformasikan ke dalam genom tanaman padi kultivar Nipponbare dengan menggunakan A. tumefaciens strain LBA 4404. Analisis molekuler dilakukan terhadap tiga anakan dari masing-masing empat rumpun tanaman Nipponbare transgenik generasi T0 (event 1, 2, 3, dan 4) dan empat kelompok rumpun tanaman T0 yang berasal dari kalus yang sama. Dari masing-masing kelompok tersebut diambil 3 rumpun T0. Hasil analisis qPCR menunjukkan bahwa anakan-anakan yang berasa...
The development of transgenic crop using Agrobacterium tumefaciens produces different inserted tr... more The development of transgenic crop using Agrobacterium tumefaciens produces different inserted transgenes, whether copy numbers or location in the plant genome. The research was performed to detect chimeric phenomena based on the transgene quantity analysis in tillers of a clump and of some clumps which were derived from the same calli. CsNitr1–L gene and hptII gene as a marker on a binary plasmid pCAMBIA1300 was transformed into the Nipponbare rice genome using A. tumefaciens strain LBA 4404. Molecular analysis was carried out on three tillers of each four clumps of Nipponbare transgenic T0 generation (events number 1, 2, 3, and 4) and four groups of T0 clump derived from one callus. Three T0 clump samples were collected from each of the four groups of T0 clumps. The results of qPCR analysis showed that the transgene copy numbers of tillers which were derived from one T0 clump were the same. qPCR analysis also discovered that not all plants from one callus demonstrated the same tra...
Cold shock protein (Csp) is an essential bacterial protein for increasing abiotic stress toleranc... more Cold shock protein (Csp) is an essential bacterial protein for increasing abiotic stress tolerance, especially cold stress. Several studies discovered that overexpression of the gene successfully improves the tolerances of several types of plant not only under cold stress, but also other abiotic stresses, e.g. hot and drought conditions. The objectives of this study were to construct a binary vector containing the LcCsp gene modified from Lactobacillus casei and transform the gene into Nipponbare rice genome. The native LcCsp gene sequence, however, has low GC content (46.7%) while rice as transformation target plant has 52% GC content. The native LcCsp gene sequence, therefore, was optimized to the level close to 52.7% similar to GC content of the rice genome. This LcCsp gene was synthesized by using DNA printing technology (gBlocks® Gene Fragments Entry, IDT). The synthetic LcCsp gene was successfully inserted into the pCAMBIA1300-int binary vector driven by Ubiquitin1 promoter an...
Pepper yellow leaf curl virus (PepYLCV) infection transmitted by silverleaf whitefly (Bemisia tab... more Pepper yellow leaf curl virus (PepYLCV) infection transmitted by silverleaf whitefly (Bemisia tabaci [Gennadius]) can decrease chili pepper yield up to 100%. At this moment, there is no chili pepper variety resistant to PepYLCV available. Genome editing approach through CRISPR/Cas9 is an effort to develop variety resistance to the viral infection. The purpose of this study was to obtain M2 lines developed by CRISPR/Cas9 system on proliferating cell nuclear antigen (PCNA) gene for resistance to PepYLCV. A total of four M2 lines (C47-7, L84-2, L84-23, and L120-19) consisting of 60 chili plants were tested for their resistance to PepYLCV. PCR analysis was performed to detect the presence (infection) of the virus. The results showed that a total of 35 plants derived from the four lines were resistant to PepYLCV. They consisted of 7 plants from C47-7 line, 11 plants from L84-2 line, 9 plants from L84-23 line, and 8 plants from L120-19 line. PCR analysis confirmed that the resistant plant...
Rice bacterial leaf blight (BLB) disease caused by Xanthomonas oryzae pv. oryzae (Xoo) affected g... more Rice bacterial leaf blight (BLB) disease caused by Xanthomonas oryzae pv. oryzae (Xoo) affected grain yield and decreasing rice production in rice growing countries. Conde, Indonesian rice variety, exhibits high resistance to most of the Indonesian races of (BLB) and has been used in Indonesia for cultivated rice. This study was aimed to conduct the molecular detection between proximal markers in chromosome 6 and relative expression of Conde rice variety compare to IRBB7 in Xa7 region. The population screening, BLB evaluation and molecular detection around the Xa7 region were conducted. The results showed from the collection of individual recombinants between resistant and susceptible parents narrow the region containing the BTBPOZ domain. The sequence alignment of Xa7LD37 in two resistant and three susceptible cultivars demonstrated a perfect association. The sequence alignment in exon region of Loc_Os06g46240 in Nipponbare, IRBB7, and IR64 identified indel/SNPs in this region lead...
THE SECOND INTERNATIONAL CONFERENCE ON GENETIC RESOURCES AND BIOTECHNOLOGY: Harnessing Technology for Conservation and Sustainable Use of Genetic Resources for Food and Agriculture
THE SECOND INTERNATIONAL CONFERENCE ON GENETIC RESOURCES AND BIOTECHNOLOGY: Harnessing Technology for Conservation and Sustainable Use of Genetic Resources for Food and Agriculture
Keragaman Jumlah Salinan Transgen CsNitr1-L pada Galur Transforman T0 Padi Kultivar Nipponbare. P... more Keragaman Jumlah Salinan Transgen CsNitr1-L pada Galur Transforman T0 Padi Kultivar Nipponbare. Perakitan tanaman transgenik dengan menggunakan bantuan Agrobacterium tumefaciens menghasilkan penyisipan transgen yang berbeda, baik dalam jumlah salinan maupun letak transgen dalam genom tanaman. Penelitian ini bertujuan untuk menganalisis keberadaan kimera dan jumlah salinan pada tiap anakan dalam satu rumpun dan beberapa rumpun padi Nipponbare transforman T0 yang berasal dari kalus yang sama. Gen CsNitr1-L pada plasmid biner pCAMBIA1300 ditranformasikan ke dalam genom tanaman padi kultivar Nipponbare dengan menggunakan A. tumefaciens strain LBA 4404. Analisis molekuler dilakukan terhadap tiga anakan dari masing-masing empat rumpun tanaman Nipponbare transgenik generasi T0 (event 1, 2, 3, dan 4) dan empat kelompok rumpun tanaman T0 yang berasal dari kalus yang sama. Dari masing-masing kelompok tersebut diambil 3 rumpun T0. Hasil analisis qPCR menunjukkan bahwa anakan-anakan yang berasa...
The development of transgenic crop using Agrobacterium tumefaciens produces different inserted tr... more The development of transgenic crop using Agrobacterium tumefaciens produces different inserted transgenes, whether copy numbers or location in the plant genome. The research was performed to detect chimeric phenomena based on the transgene quantity analysis in tillers of a clump and of some clumps which were derived from the same calli. CsNitr1–L gene and hptII gene as a marker on a binary plasmid pCAMBIA1300 was transformed into the Nipponbare rice genome using A. tumefaciens strain LBA 4404. Molecular analysis was carried out on three tillers of each four clumps of Nipponbare transgenic T0 generation (events number 1, 2, 3, and 4) and four groups of T0 clump derived from one callus. Three T0 clump samples were collected from each of the four groups of T0 clumps. The results of qPCR analysis showed that the transgene copy numbers of tillers which were derived from one T0 clump were the same. qPCR analysis also discovered that not all plants from one callus demonstrated the same tra...
Cold shock protein (Csp) is an essential bacterial protein for increasing abiotic stress toleranc... more Cold shock protein (Csp) is an essential bacterial protein for increasing abiotic stress tolerance, especially cold stress. Several studies discovered that overexpression of the gene successfully improves the tolerances of several types of plant not only under cold stress, but also other abiotic stresses, e.g. hot and drought conditions. The objectives of this study were to construct a binary vector containing the LcCsp gene modified from Lactobacillus casei and transform the gene into Nipponbare rice genome. The native LcCsp gene sequence, however, has low GC content (46.7%) while rice as transformation target plant has 52% GC content. The native LcCsp gene sequence, therefore, was optimized to the level close to 52.7% similar to GC content of the rice genome. This LcCsp gene was synthesized by using DNA printing technology (gBlocks® Gene Fragments Entry, IDT). The synthetic LcCsp gene was successfully inserted into the pCAMBIA1300-int binary vector driven by Ubiquitin1 promoter an...
Pepper yellow leaf curl virus (PepYLCV) infection transmitted by silverleaf whitefly (Bemisia tab... more Pepper yellow leaf curl virus (PepYLCV) infection transmitted by silverleaf whitefly (Bemisia tabaci [Gennadius]) can decrease chili pepper yield up to 100%. At this moment, there is no chili pepper variety resistant to PepYLCV available. Genome editing approach through CRISPR/Cas9 is an effort to develop variety resistance to the viral infection. The purpose of this study was to obtain M2 lines developed by CRISPR/Cas9 system on proliferating cell nuclear antigen (PCNA) gene for resistance to PepYLCV. A total of four M2 lines (C47-7, L84-2, L84-23, and L120-19) consisting of 60 chili plants were tested for their resistance to PepYLCV. PCR analysis was performed to detect the presence (infection) of the virus. The results showed that a total of 35 plants derived from the four lines were resistant to PepYLCV. They consisted of 7 plants from C47-7 line, 11 plants from L84-2 line, 9 plants from L84-23 line, and 8 plants from L120-19 line. PCR analysis confirmed that the resistant plant...
Rice bacterial leaf blight (BLB) disease caused by Xanthomonas oryzae pv. oryzae (Xoo) affected g... more Rice bacterial leaf blight (BLB) disease caused by Xanthomonas oryzae pv. oryzae (Xoo) affected grain yield and decreasing rice production in rice growing countries. Conde, Indonesian rice variety, exhibits high resistance to most of the Indonesian races of (BLB) and has been used in Indonesia for cultivated rice. This study was aimed to conduct the molecular detection between proximal markers in chromosome 6 and relative expression of Conde rice variety compare to IRBB7 in Xa7 region. The population screening, BLB evaluation and molecular detection around the Xa7 region were conducted. The results showed from the collection of individual recombinants between resistant and susceptible parents narrow the region containing the BTBPOZ domain. The sequence alignment of Xa7LD37 in two resistant and three susceptible cultivars demonstrated a perfect association. The sequence alignment in exon region of Loc_Os06g46240 in Nipponbare, IRBB7, and IR64 identified indel/SNPs in this region lead...
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