Jurnal AgroBiogen
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Gen dan QTL Pengendali Toleransi Tanaman terhadap Keracunan Aluminium dan Aplikasinya untuk Pemuliaan Tanaman di Indonesia
Get PDFGenetic knowledge of loci controlling Al toxicity tolerance is the key for a successful breeding program in developing Altolerant cultivars. Tolerance level of crop plants to Al toxicity is genetically controlled. The gene inheritance pattern is mainlyresulted from intensive studies of cereal crops, such as wheat, sorghum, maize, and rice. The trait can be controlled by asingle dominant gene, a single dominant gene with many alleles, a pair of dominant genes, or by many genes (QTL). Themajority of the Al tolerance genes identified so far belongs to two independent groups of gene families, i.e. aluminumactivatedmalate transporter (ALMT) and multidrug and toxic compound extrusion (MATE), both encoding transport proteinsinvolved in Al-activated organic acid release, mainly citrate and malate. The variations in Al toxicity tolerance phenotypes arestrongly correlated with the expressions of such genes in the root apical cells. Many Al tolerance QTLs have been mapped inthe genomes of various crop species and were found to be colocated with the ALMT and MATE genes. The genetic maps ofthe Al tolerance genes and QTLs facilitate breeding programs for developing Al-tolerant cultivars through marker-assistedbreeding methods. Al tolerance genes that have been isolated from genetically unrelated species can be used in genetictransformation studies of crop genotypes sexually incompatible to the gene source genotypes. The application of thesemolecular breeding methods expedites breeding programs to develop crop cultivars tolerance to Al toxicity and acid soils.Genomic technologies by using next-generation sequencing and high-throughput genotyping system accelerate Al toxicitytolerance gene and QTL discoveries of various crop species. The modern genomic technologies also facilitate morecomprehensive PGR characterization and utilization to accelerate identification and isolation of the Al tolerance genes andQTLs to be used in a more comprehensive breeding program to support national food self sufficiency and food securityprograms
Keragaman Somaklonal untuk Perbaikan Tanaman Artemisia (Artemisia annua L.) melalui Kultur In Vitro
Get PDFSomaclonal Variability for the Improvement of PlantsArtemisia (Artemisia annua L.) by In Vitro Culture.Endang G. Lestari, Rosa Yunita, and Ali Husni. Artemisiaannua L., a family member of Asteraceae, is medicinalplants originated from China. The plant has been widelyused by the local people for malaria remedy. Its active substance,artemisine, has been proved to hamper the malariabacteria incubation, Plasmodium sp. In accordance with theWHO recomendation, the Department of Health of Indonesiais now in the attempt of developing this plant as thesubtitute of chloroquin because of the malaria bacteriaresistance to this antidote. In Indonesia, the artemisinecontent of the plant less than 0,5% is the crucial problemleading no investors are interested in its economic value.Therefore, Indonesian Medicinal and Spice Crops ResearchInstitute; BPTO Tawangmangu, Indonesian Institute ofSciences; and PT Kimia Farma cooperate for obtaining theprime clone by breeding, selection, as well as environmentaladaptation. In coping with the problem, ICABIOGRAD in thecollaboration with Bogor Agricultural University haveconducted the research for genetic improvement throughmutative induction and field selection. This research onsomaclonal variation. was conducted from Januari 2006 toJuni 2008. Eksplan used for experiment were shoots radiatedwith 10-100 Gy gamma ray. The result showed that the shootradiated with the dosage of 70-100 Gy was unable to grow.On the other hand, the high level of multiplication wasacquired in the one radiated with 10-30 Gy. The optimumradiation for somaclonal radiation was eventually gainedwith 40-60 Gy. The somaclone lines with 10-60 Gy radiationhave been aclimatized and planted in Gunung Putri plot inthe elevation of 1545 asl. Artemisinin content at the highbiomases genotype is 0,49-0,52%
Construction and Expression of Pet Operon using Shuttle Vector for Mesophilic and Thermophilic Bacteria
Get PDFKeuntungan fermentasi etanol pada suhu tinggi mendorong penelitian perakitan bakteri termofilik etalogenik. Selain itu, kemampuan bakteri termofilik dalam penggunaan gula pentosa hasil degradasi biomasa memberi peluang untuk menekan biaya produksi bioetanol. Tujuan dari penelitian ini adalah untuk mengkonstruksi pet (production of ethanol) operon dengan menggunakan shuttle vector pMK18 dan melihat ekspresinya dalam bakteri mesofilik dan termofilik. Konstruksi dan ekspresi pet operon dengan menggunakan adhT dari bakteri termofilik dan pdc dari bakteri mesofilik, dan penggunaan mesofilik-termofilik shuttle vector sebagai backbone-nya baru pertama kali dilaporkan. Pet operon adalah suatu susunan gen penyandi produksi etanol yang terdiri dari gen pdc (pyruvate decarboxylase) dan adh (alcohol dehydrogenase). Konstruksi pet operon menggunakan gen adhT dari bakteri termofilik Geobacillus thermoglucosidasius M10EXG dan pdc (pyruvate dehydrogenase) dari bakteri mesofilik Zymomonas mobilis ZM4 telah dilakukan dengan menggunakan mesofilik-termofilik shuttle vector pMK18. Ekspresi pet operon pada bakteri mesofilik Eschericia coli dapat memproduksi 0,3 g/l etanol dengan aktivitas adhT sekitar 0,02 U/mg protein dan aktivitas pdc sekitar 0,004 U/mg protein. Perlu dilakukan penelitian lanjutan untuk perbaikan konstruksi pet operon untuk sistemtermofik pada Thermus thermophilus HB27, karena konstruksi yang didapat belum optimum untuk sistem termofilik ini. Hasil ini diharapkan akan mengawali pengembanganteknik manipulasi genetik pada bakteri termofilik yang masih sangat terbatas, khususnya pengembangan teknik manipulasi termofilik etanologenik. Kata kunci: Etanol, bakteri termofilik, bakteri mesofilik, pet operon, ekspresi gen.
Perbanyakan Nematoda Patogenik Serangga (Rhabditida: Steinernema dan Heterorhabditis) pada Media In Vitro Cair Statik
Get PDFEntomopathogenic nematodes belongingto genera Steinernema and Heterorhabditis are potentiallymost effective and safe biological control agents for insectpests, especially for soil dwelling insects and those living incryptic habitats. Field application of the nematodes is stillhampered by supply of large number of infective juvenile(IJ) nematodes. Five published in vitro media along with itstwo modifications were tested for mass propagations of twoindigenous nematodes (H. indicus PLR2 and SteinernemaT96) and one commercial strain (S. carpocapsae #25).Varying levels of IJ yields were observed across thereplications and experiments. Medium F that contained 1.0%yeast extract, 2.5% egg yolk, and 4.0% soy oil yielded thehighest IJ numbers of H. indicus PLR2 (1.5×105 IJ ml-1) andof S. carpocapsae #25 (2.9×105 IJ ml-1), whereas the widelyused medium B, which is based on homogenized chickenoffal (40%), yielded the highest number of Steinernema T96(5.8×104 IJ ml-1). The IJ’s quality, as measured by theirmorphometrics and pathogenicities, were generallyimpaired, indicating the lack of essential nutrient(s) in themedia. Optimization of the propagation media is thereforestill needed to increase IJ’s quantity and quality to achievethe required standard for commercial scale of artificialpropagation
Keragaman Genetik Isolat Cendawan Pyricularia oryzae Menggunakan Primer Pot-2 (Rep-PCR)
Get PDFRice blast (Pyriculariaoryzae) is one of the most important diseases of rice. It canbe very destructive in the field, when the environmentalconditions are favourable. Information on genetic diversity ofthis pathogen could assist plant breeders in determiningstrategy for a successful control of the disease. This studywas conducted to analyze genetic diversity in P. oryzaeisolates by a pair of Pot-2 primers using the rep-PCRtechnique. These primers were designed from a transposonelement of the entire blast fungus genomic DNA. DNAsamples were extracted from 212 isolates of P. oryzaecollected from two endemic areas of the disease inIndonesia, i.e., Tamanbogo, Lampung, and Sukabumi, WestJava, as well as from some non-endemic areas in NorthSumatra and West Sumatra). Results of the study indicatedthat the 212 isolates could clustered into 21 haplotypes. Themost dominant haplotypes as indicated by their highestfrequency of haplotypes were haplotype Pot 2-019 (54.46%)followed by haplotype Pot 2-021 (14.73%) and haplotipe Pot2-016 (6.25%). Regardless of origins of the P. oryzae isolates,we found 6 haplotypes from Tamanbogo (out of 117samples), 13 haplotypes from Sukabumi (out of 77 samples),and 11 haplotypes from North Sumatra and West Sumatra(out of 18 isolates). It seems that genetic diversity of the P.oryzae isolates was not affected by the total number ofsamples/isolates, but rather by place of the origin and ricegenotypes from which the isolates were collected
Konstitusi Genetik dan Karakter Fenotipik Galur-galur Padi Pup1 Turunan Varietas Situ Bagendit
Get PDFAcidity, phosphorus deficiency, and droughtstress are major problems in Indonesia’s Ultisol rice farming.Development of rice lines tolerant to those stresses isexpected to be able to reduce the consumption of Pfertilizer. The objectives of the research were to evaluategenetic constitutive of rice lines (BC2F6 population) derivedfrom Situ Bagendit x Kasalath and Situ Bagendit x NIL-C433crossings, and to evaluate responses of those lines toYoshida nutrient solution under P deficiency and Al stresscondition. The research was conducted at Molecular BiologyLaboratory and Greenhouse of ICABIOGRAD, fromNovember 2011 to May 2013. The result of foregroundanalysis showed that Pup1 locus has been integrated intothe genome of BC2F6 rice lines, eventhough some lines (SK5,SK6, SK7, SK8, SK9, SK10, SK19, and SK20) showedincomplete integration. Background analysis indicated thatmajority (95.7%) of the Situ Bagendit background has beenrecovered in BC2F6 rice lines. Al stress evaluation showed SNlines were more tolerant to P deficiency and Al stress thanthat of SK lines. Pup1 locus showed good expression underlow P and no Al stress. Based on genome proportion andYoshida nutrient solution experiments, a total of three lines,namely SK13, SN2, and SN9, have potential goodcharacteristics. Molecular analysis within a marker-assistedbackcrossing (MAB) experiment should be carried out ateach generation of lines for gaining fully gene segment inadvanced generations
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)
Get PDFBreeding 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
TINJAUAN Penggunaan Suspensi Sel dalam Kultur In Vitro
Get PDFCell suspension culture could be defined as aprocess that allows rapidly dividing homogenous suspensionof cells to grow in liquid nutrient media. There are two maintypes of suspension cultures: (1) Batch cultures in whichcells are nurtured in a fixed volume of medium until growthceases and (2) Continuous cultures in which cell growth ismaintained by continuous replenishment of sterile nutrientmedia. Plant cell suspension cultures are mostly used for thebiochemical investigation of cell physiology, growth, metabolism,protoplast fusion, transformation and for large scaleproduction of seed by bioreactor and production of secondarymetabolites. Contamination is one of the largest problemswhen dealing with cell cultures. Differences betweenthe products of cell suspension culture and whole plant arefrequently observed. These phenomena’s may be resultedfrom lack of differentiation and organization and cell cultureinducedvariation. Utilization of cell suspension culture inIndonesia is still limited, some of them for mass productionof plantation seed with bioreactor system and for productionof secondary metabolites. The success of this study give theopportunity for mass production of seeds from other plantsand also production of secondary metabolites
Identifikasi Perubahan Karakter Agronomis Padi Transgenik Penanda Aktivasi cv. Asemandi Generasi T1
Get PDFThe activation-tagged populations of transgenic rice cv.Asemandi have been developed by introducing construct ofactivation taq Ac/Ds into rice genom of cv. Asemandi. The T1transgenic rice populations cv. Asemandi containingconstruct of activation taq have been obtained and neededto be characterized. This study aimed to identify the Bastaherbicide resistant plants, transgenic plants containing hptand bar genes, and changes in agronomic traits. Bastaresistant plant was identified by treating leaf with bastasolution. Hpt and bar genes were detected by PCR usingspecific primers. Phenotype characters were identified byobserving and measuring their agronomic parameters. Thestudy results showed that out of 315 rice transgenic cv.Asemandi T1 treated with Basta solution, 176 (55.87%) plantswere indicated to be resistant to Basta. The results of PCRanalysis revealed that eight rice transgenic cv. Asemanditested contained both hpt and bar genes. In general,compared to the nontransgenic plants, there were changesin several agronomic parameters of T1 transgenic plants cv.Asemandi, including plant height, days to flowering, days toharvesting, periods of grain filling, and weight of 100 grains.Correlation analysis showed that there was no correlationbetween days of harvesting to weight of 100 grains intransgenic rice, but there was correlation in nontransgenicrice. Transgenic rice plants cv. Asemandi with changes inthe agronomic characters will be useful for further study,such as to analyze the function of the genes
Induksi Kalus dan Regenerasi Beberapa Genotipe Gandum (Triticum aestivum L.) secara In Vitro
Get PDFCallus Induction and In Vitro Plant Regeneration ofWheat Genotypes (Triticum aestivum L.). AtmitriSisharmini, Aniversari Apriana, and Sustiprijatno. Developmentof a reliable in vitro plant regeneration procedure forwheat is a prerequisite for its improvement by genetic transformation.The purpose of this study was to obtain methodsof callus induction and regeneration of wheat genotypes.This experiment was conducted at ICABIOGRAD. Immatureembryos from four wheat genotypes, ie Perdix, Naxos Wew,Combi and Fasan were used to induce callus formation andregeneration rate of callus. For the preparation of callusinduction medium, MS-L7 basal medium was supplementedwith combination of growth regulators 2,4 dichlorophenoxyacetic acid (2,4-D) and 4-amino-3,5,6-trichloropicolinic acid(picloram). While, plant regeneration medium was preparedusing MS basal medium supplemented with combination ofthree growth regulators i.e. IAA, BAP and kinetin. The resultsshowed that genotype, in vitro culture medium and growthregulators played a dominant role in callus induction andplantlet regeneration. All the 4 genotypes responded positivelyto callus induction, however, variability was observednot only among the genotypes but also within callusinduction medium used. The best induction medium wasthe MS-L7 basal medium supplemented with combination ofphytohormon 4 mg/l 2,4-D + 2 mg/l picloram (GIK-3) whichshowed 100% callus induction frequency. Whereas, the bestregeneration medium was shown by MS basal medium withcombination of phytohormon 1.5 mg/l BAP dan 0.5 mg/lkinetin (RG3). Regarding plant regeneration, Perdix was themost responsive genotype to be regenerated with regenerationfrequency of 57.33%. The successfully acclimatizedplanlets in greenhouse were obtained from Perdix andNaxos Wew genotypes. These results will potentially facilitategenetic transformation research of wheat in Indonesia