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Introduksi Konstruk Gen CsNitr1-L dengan Promotor Ubiquitin melalui Agrobacterium tumefaciens dan Deteksi Molekulernya pada Padi Kultivar Nipponbare Enngarini, Wening; Polosoro, Aqwin; Sustiprijatno, Sustiprijatno; Trijatmiko, Kurniawan Rudi
JURNAL BIOLOGI INDONESIA Vol 13, No 2 (2017): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v13i2.3400

ABSTRACTNitrogen based fertilizers such as urea and NPK are primary needs for rice farmers. To get significant improvement of crop yield, the more quantity of fertilizers are applied. It make negative impact for surrounding environment. Based on that, the efforts should be done to suppress the demand of fertilizers such as by developing Nitrogen Use Efficiency crops. CsNitr1-L is one of gene that related to Nitrogen Use Efficiency trait in plant. The objectives of this research are to develop the construction of CsNitr1-L gene candidate in pCAMBIA1300-Ubi1 promoter and to obtain the transformants of rice cultivar Nipponbare which contain the construction of CsNitr1-L gene candidate. The construction of pCAMBIA1300::Ubi1::CsNitr1-L has successfully assembled and was transformed to immature embryo of rice cultivar Nipponbare using Agrobacterium tumefaciens strain LBA4404. It was obtained 146 lines of T0 Nipponbare. PCR analysis of T0 Nipponbare lines showed that 66 of them was identified as positive T0 lines contained hptII and CsNitr1-L genes. Transformation efficiency obtained was 11,9%. The result of analysis copy number using Southern Hybridization in positive PCR of T0 lines randomly indicated that 4 lines have a single copy of transgene. Based on these results, it can be concluded that CsNitr1-L gene construct was successfully introduced into the genome of the rice plant cultivar Nipponbare and the positive PCR of T0 lines containing the gene of hptII and CsNitr1-L, also a single copy of the transgene was obtained.Keywords: Nitrogen use efficiency trait, gene construction, rice kultivar Nipponbare

Molecular Evaluation for Drought Tolerant Using Marker Assisted Breeding Method Fatimah, Fatimah; Prasetiyono, Joko; Trijatmiko, Kurniawan Rudi; Sustiprijatno, Sustiprijatno
ANNALES BOGORIENSES Vol 22, No 2 (2018): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ann.bogor.2018.v22.n2.94-100

The sustainability and increasing the national rice production require the readiness of food and agriculture sector cope with the impacts of climate change, land degradation, drought area, sloping production and the raising of population growth. Adaptation plays an important role in ensuring the sustainability of food security. This research aimed to develop drought-tolerant variety of Inpari 30 (submergence tolerance variety) and Situ Bagendit through marker-assisted backcrossing-through pyramiding gene of identified QTLs for foreground selection and to explore SSRs and 6K SNPs for background selection distributed in 12 rice chromosome of drought tolerant donor (Cabacu) and recipient rice (Inpari 30 and Situ Bagendit). The foreground selection revealed that flanking SSRs of each QTLs (qRPF2.1, qGPP2.1, qSPP4.1 and Sub1) was less than 2 cM. The background selection through polymorphic survey of Rice 6K SNP primers revealed 2457 (53,3%) polymorphic SNPs on Inpari 30 vs Cabacu and 2563 (55,6%) polymorphic SNPs on Situ Bagendit vs Cabacu with the average distance about 0.74 cM/chromosome. The genotypic selection of F1 Inpari 30/Cabacu and F1 Situ Bagendit/Cabacu have already in heterozygote condition for these 4 QTLs target. These lines was continued for backcross breeding to develop BC1F1 Inpari 30/Cabacu and BC1F1 Situ Bagendit/Cabacu generation.

EVALUASI KEMUDAHAN TRANSFER MARKA SSR PADI UNTUK MENGANALISIS KERAGAMAN GENETIK FAMILI POACEAE TOLERAN KEKERINGAN Fatimah, Fatimah; Masumah, Masumah; Prasetiyono, Joko; Sustiprijatno, Sustiprijatno
JURNAL BIOLOGI INDONESIA Vol 15, No 1 (2019): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v15i1.3764

ABSTRACTAnticipating climate change and its impacts on crop production, it requires the development of adaptive rice varieties that respond to drought stress and minimize the damage while keeps growing and maintaining yield. The aim of this study was to evaluate the transferability of rice simple sequence repeat (SSR) markers to Poaceae family and analyze the genetic diversity among 16 accessions of Poaceae family (12 genera and 14 species) using 41 rice SSR markers. The result revealed that the transferability of rice SSR markers was varied. High amplification produced in rice group (>70%) and low amplification in grass group (<30%) with an average of 35.2%. A total of 128 cross-species alleles were identified with an average of 3 alleles/locus. The value of gene diversity ranged from 0.15 to 0.83 with an average of 0.53 and the value of Polymorphic Information Content (PIC) ranged from 0.14 to 0.80 with an average of 0.46. The results of phylogenetic analysis determined two clusters at similarity coefficient of 0.72. The first cluster consisted of 14 accessions from Poaceae family (12 genera and 14 species) while the second cluster consisted of two cultivated rice varieties (Inpari 30 and Situ Bagendit). The genetic relatedness data revealed from this study could be used as basic information for parental selection. The 14 accessions of Poaceae family have a potential for drought tolerant donor and separated clearly from cultivated rice varieties (Inpari 30 and Situ Bagendit) for recipient parents in spike-stalk injection method (SIM) to develop drought tolerant rice varieties. Keywords: Cross amplification, Drought tolerant, Inpari 30, Situ Bagendit

EVALUASI KEMUDAHAN TRANSFER MARKA SSR PADI UNTUK MENGANALISIS KERAGAMAN GENETIK FAMILI POACEAE TOLERAN KEKERINGAN Fatimah, Fatimah; Masumah, Masumah; Prasetiyono, Joko; Sustiprijatno, Sustiprijatno
JURNAL BIOLOGI INDONESIA Vol 15, No 1 (2019): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v15i1.3764

ABSTRACTAnticipating climate change and its impacts on crop production, it requires the development of adaptive rice varieties that respond to drought stress and minimize the damage while keeps growing and maintaining yield. The aim of this study was to evaluate the transferability of rice simple sequence repeat (SSR) markers to Poaceae family and analyze the genetic diversity among 16 accessions of Poaceae family (12 genera and 14 species) using 41 rice SSR markers. The result revealed that the transferability of rice SSR markers was varied. High amplification produced in rice group (>70%) and low amplification in grass group (<30%) with an average of 35.2%. A total of 128 cross-species alleles were identified with an average of 3 alleles/locus. The value of gene diversity ranged from 0.15 to 0.83 with an average of 0.53 and the value of Polymorphic Information Content (PIC) ranged from 0.14 to 0.80 with an average of 0.46. The results of phylogenetic analysis determined two clusters at similarity coefficient of 0.72. The first cluster consisted of 14 accessions from Poaceae family (12 genera and 14 species) while the second cluster consisted of two cultivated rice varieties (Inpari 30 and Situ Bagendit). The genetic relatedness data revealed from this study could be used as basic information for parental selection. The 14 accessions of Poaceae family have a potential for drought tolerant donor and separated clearly from cultivated rice varieties (Inpari 30 and Situ Bagendit) for recipient parents in spike-stalk injection method (SIM) to develop drought tolerant rice varieties. Keywords: Cross amplification, Drought tolerant, Inpari 30, Situ Bagendit

INTRODUKSI KONSTRUK GEN CSNITR1-L DENGAN PROMOTOR UBIQUITIN MELALUI AGROBACTERIUM TUMEFACIENS DAN DETEKSI MOLEKULERNYA PADA PADI KULTIVAR NIPPONBARE Enngarini, Wening; Polosoro, Aqwin; Sustiprijatno, Sustiprijatno; Trijatmiko, Kurniawan Rudi
JURNAL BIOLOGI INDONESIA Vol 13, No 2 (2017): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v13i2.3400

ABSTRACTNitrogen based fertilizers such as urea and NPK are primary needs for rice farmers. To get significant improvement of crop yield, the more quantity of fertilizers are applied. It make negative impact for surrounding environment. Based on that, the efforts should be done to suppress the demand of fertilizers such as by developing Nitrogen Use Efficiency crops. CsNitr1-L is one of gene that related to Nitrogen Use Efficiency trait in plant. The objectives of this research are to develop the construction of CsNitr1-L gene candidate in pCAMBIA1300-Ubi1 promoter and to obtain the transformants of rice cultivar Nipponbare which contain the construction of CsNitr1-L gene candidate. The construction of pCAMBIA1300::Ubi1::CsNitr1-L has successfully assembled and was transformed to immature embryo of rice cultivar Nipponbare using Agrobacterium tumefaciens strain LBA4404. It was obtained 146 lines of T0 Nipponbare. PCR analysis of T0 Nipponbare lines showed that 66 of them was identified as positive T0 lines contained hptII and CsNitr1-L genes. Transformation efficiency obtained was 11,9%. The result of analysis copy number using Southern Hybridization in positive PCR of T0 lines randomly indicated that 4 lines have a single copy of transgene. Based on these results, it can be concluded that CsNitr1-L gene construct was successfully introduced into the genome of the rice plant cultivar Nipponbare and the positive PCR of T0 lines containing the gene of hptII and CsNitr1-L, also a single copy of the transgene was obtained.Keywords: Nitrogen use efficiency trait, gene construction, rice kultivar Nipponbare

Characterization of a Drought-Inducible Dehydrin Promoter from Sugarcane (Saccharum officinarum L.) in Tobacco (Nicotiana tabacum L.) Iskandar, Hayati Minarsih; Suhandono, Sonny; Pambudi, Jembar; Kristianti, Tati; Putranto, Riza Arief; Mose, Windi; Sustiprijatno, Sustiprijatno
Journal of Tropical Crop Science Vol 7 No 01 (2020): Journal of Tropical Crop Science
Publisher : Department of Agronomy and Horticulture, IPB University

Dehydrin (DHN) is known to play an important role in plant response and adaptation to abiotic stresses (drought, high salinity, cold, heat, etc.). Previous research reported the increased expression of DHN in sugarcane stems exposed to drought stress for 15 days which may be controlled by its corresponding stress inducible promoter. The DHN promoter was succesfully isolated from sugarcane variety PSJT 941 (Pr-1DHNSo) and was cloned to pBI121 expression vector fused to a β-glucuronidase (GUS) reporter gene. The aim of this research was the functional testing of the Pr-1DHNSo promoter through transformation into tobacco plant treated with in vitro drought stress. Genetic transformation of Pr-1DHNSo construct was conducted by Agrobacterium tumefaciens. The transformed tobacco was then subjected to drought stress treatment using 40% PEG 6000 for five sequential incubations (0, 12, 24, 48 and 72 hours). The GUS assay reveal that the transformed tobacco treated with drought stress showed a blue color denoting GUS activity in leaf, stem and root tissues and this expression increased along with the length of the drought treatment. The analysis of gusA gene using real time-qPCR normalized to the L25 reference gene also showed that the expression increased in line with the length of time of drought stress. The results presented in this study indicated that the Pr-1DHNSo promoter from sugarcane was expressed and induced by drought stress treatment in tobacco.

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