Identifikasi Galur dan Gen-gen Terkait Toleran Kekeringan pada Padi Transgenik cv. T309 yang Mengandung Vektor Penanda Aktivasi

Activation tagging is an efficient tool forfunctional analysis of the rice genes. We have developed anumber of transgenic rice lines (Oryza sativa L. ssp.japonica cv. Taipei 309) containing activation tagging vector.However, the phenotypes and genotypes of these lines, inrelation to the drought stress, have not been analyzed. Theobjectives of this research were to identify transgenic ricelines that showed tolerance to the drought stress and toidentify the genes that may be associated with the droughtstress. The drought stress tolerance in transgenic rice lineswas identified by testing their tolerance to the drought stressand also by detecting the presence of bar and nptII genes.The result showed that 56 out of 59 rice lines were resistantto Basta herbicide and three of them showed tolerance todrought stress, namely PA.T-1.2, PA.T-4.1, and PA.T-5.1 lines.PCR analysis showed that PA.T-1.2 and PA.T-4.1 containedboth hptII and bar genes, while the PA.T-5.1 line containedbar gene only. Thermal Asymetric Interlaced-PCR (TAILPCR)analysis showed that two genes may be asssociatedwith the drought stress tolerance. Those genes areOSJNBa0004120.14 that produces uridylate putative kinaseand OsPPCK2L that produces phosphoenolpyruvatecarboxylase kinase

Pengembangan Populasi Mutan Penanda Aktivasi: I. Transformasi Padi Japonica Tropis Lokal Sulawesi CV. Asemandi Dengan Bantuan Agrobacterium Tumefaciens

The rice transformation technologyis not only provides valuable methods for the introductionof useful genes into rice plant to improve importantagronomic traits, but also helps in studying gene functionand regulation based on rice genome sequence information.Knockout of genes by insertional mutagenesis is a straightforwardmethod to identify gene functions. One of themethods to develop rice mutants is through genetic transformationmediated by Agrobacterium using activationtagging by Ac-Ds system. A study was done with an objectiveto obtain mutant rice of local tropical japonica cv. Asemandithrough genetic trans-formation mediated by Agrobacteriumtumefaciens. The transformation was conducted usingAgrobacterium vector with the strain of Agl-1 containingactivation tag construct. The result of experiment showedthat it has been obtained 17 independent line (304 plants)transgenic Asemandi containing activation tag construct.These starter lines will be used as materials to developseveral generations of stabil rice mutant through selfing

Introduksi Konstruk Gen CsNitr1-L dengan Promotor Ubiquitin melalui Agrobacterium Tumefaciens dan Deteksi Molekulernya pada Padi Kultivar Nipponbare

Nitrogen 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

Deteksi Gen HptII dan Keragaan Agronomis pada Populasi BC1F1 Tanaman Padi Transgenik

Rice varieties tolerant to drought stress are needed tostabilize rice production under drought stress condition. Wedeveloped transgenic rice cv. Nipponbare carrying hptIIgene that might also contain OsDREB1A gene. OsDREB1Agene responsible to drought tolerance trait need to betransferred into cultivated rice in order to obtain new localrice variety tolerant to drought stress. The aims of thisresearch were to detect the presence of hptII gene in the F1and BC1F1 transgenic rice and to observe the agronomicperformace of those populations and their plant physiology.F1 population was developed by crossing transgenicNipponbare, as donor parent, with Batutegi, Code, Ciherang,and Konawe genotypes, as recipient parents. BC1F1population was developed by backcrossing F1 transgenicline with recipient parents, respectively. The presence ofhptII gene was analyzed by PCR using a pair of primers forhptII. The observation of agronomic performance wascarried out in the green house, meanwhile the observationof stomata was done using microscope. The result of PCRanalysis showed that BC1F1 Batutegi trans, BC1F1 Code trans,BC1F1 Konawe trans1, BC1F1 Konawe trans3, dan BC1F1Konawe trans4 were detected carrying the hptII gene.Agronomic data showed that BC1F1 transgenic rice linesyielded panicles, filled grains, and total grains higher thanthose of recipent parents. Comparing to the recipientparents, BC1F1 Konawe trans1 and BC1F1 Konawe trans3 hadless stomata on the lower side of the leaf, but had morestomata on the upper side of the leaf

Respon Padi Transgenik CV. Nipponbare Generasi T1 Yang Mengandung Gen Oryza Sativa Dehydration-response Element Binding 1a (Osdreb1a) Terhadap Cekaman Salinitas [Response of T1 Generation Transgenic Rice CV. Nipponbare Containing an Oryza Sativa Dehydration-response Element Binding 1a (Osdreb1a) Gene to Salinity Stress]

Salinity is one of the abiotic constraints in the cultivation of rice crop. One of the reasons agricultural land becomes saline is due to the intrusion of seawater into the mainland as a result of global climate change. Dehydration-responsive element binding (DREB) gene is a plant -specific transcription factor gene that have important role in regulating plant responses to abiotic stresses, including high salinity. Transgenic rice plants cv. Nipponbare carrying OsDREB1A gene have been generated. However, study of the response of putative transgenic plants to salinity has not been done. The research objective is to study the response of T1 generation Nipponbare-OsDREB1A transgenic rice plants to salinity stress. The result showed that the response of putative transgenic rice Nipponbare-OsDREB1A to salinity stress 25 mM and 150 mM NaCl indicated a level of tolerance varies from highly sensitive to highly tolerance. These variations were possibly occurred because of the segregation state of the T1 generation transgenic rice. Based on damage symptom scoring and PCR analysis provided information that transgenic rice plant cv. Nipponbare-OsDREB1A which showed positive PCR had a very high tolerance to salinity stress 150 mM compared with non-transgenic rice cv. Nipponbare

Analisis Molekuler dan Keragaan Agronomis Galur-galur Padi BC1F1 Persilangan Code x qTSN4 dan Code x qDTH8 (Molecular Analysis and Agronomic Performance of BC1F1 Crosses Code x qTSN4 and Code x qDTH8)

Breeding based on molecular marker has become a routine activity in the current rice research. The development of an earlymaturity of rice variety with high yield is needed to increase national rice production. This study aimed to determine the patternof alleles for loci controlling total spikelet number and number of days to heading, as well as agronomic performances of theBC1F1 Code x qTSN4 and Code x qDTH8 populations. The study was conducted at the Indonesian Center for Biotechnology andGenetic Resources Research and Development from January to August 2014. The plant materials used were Code (a nationalvariety with bacterial blight resistance gene [Xa7]), IR64-Nils-qTSN4[YP9] (qTSN4 that contains a locus controlling the number ofspikelet), IR64-Nils-qDTH8[YP1] (qDTH8 that contains a locus controlling the number of days to heading), BC1F1 Code x qTSN4,and BC1F1 Code x qDTH8. A total of 250 BC1F1 plants of each crosses were selected using molecular markers of RM20582 for Xa7gene, RM17483 and RM6909 for QTL position of qTSN4, RM5556 and RM6838 for QTL position of qDTH8. Based on molecularanalysis, there were 63 BC1F1-qTSN4 lines and 65 BC1F1-qDTH8 lines showing heterozygote alleles for qTSN4 or qDTH8 loci andwere homozygote for Xa7 locus (HHA pattern). Five plants from each locus target were backcrossed to the recurrent parent,Code, to obtain BC2F1 seeds. The remaining BC1F1 plants were self-pollinated to obtain BC1F2 seeds. Observations on someagronomic characters demontrated that the BC1F1 plants showed higher yield potential than Code and the flowering time of theBC1F1 progenis were also earlier than Code. These results indicated that the yield potential of Code could be improved byintrogression of qTSN4 and qDTH8 loci into the Code genome