Jurnal AgroBiogen
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Keterpautan 23 Marka Mikrosatelit pada Kromosom 6 dan 7 dengan Karakter Ketahanan Populasi Jagung terhadap Penyakit Bulai (Peronosclerospora maydis)
Get PDFLinkage of 23 Microsatellite Marker on Chromosome 6and 7 to Downy Mildew Resistance on Maize. Roberdi,Hajrial Aswidinnoor, Asep Setiawan, Sutrisno, Marcia B.Pabendon, and M. Azrai. Downy mildew caused byPeronosclerospora is one of most important maize diseasein several countries, including Indonesia. Parental andprogenies selection based on conventional breeding is timeconsuming and laborious. Development of molecularbiology produces many DNA markers used for selection, oneof them is microsatellite. The aim of this research to identifymicrosatellite markers associated with downy mildewresistance on maize progeny MR-4 X AMATLCOHS-9-1-1-1-1-1-2-B, on chromosome 6 and 7. This research was consistedof two activities, phenotypic and genotypic analysis.Phenotypic analysis used 175 progenies BC1F2 and both ofparents. This analysis included planting of spreading row,inoculums preparation, inoculation of spreader rows, testmaterial planting, inoculation of test material andobservation. Genotypic analysis used 175 progenies BC1F1and both of parents. This analysis included DNA genomeisolation, PCR analysis, electrophoresis, gel staining and datascoring. Percentage of downy mildew infections on MR-4was 76%, while these on AMATLCOHS-9-1-1-1-1-1-2-B was16%, and on 175 progenies had range from 10.1-100%. Out of23 SSR, 12 markers could be mapped in chromosome 6 and11 markers in chromosome 7. QTL analyses showed thatchromosome 7 contain one QTL in position between phi082and phi116I marker as far as 18.6 cM with 2.6 LOD value
Spektrum Ketahanan Galur Haploid Ganda Turunan IR64 dan Oryza rufipogon yang mengandung QTL Ketahanan terhadap Penyakit Blas (Pir)
Get PDFResistance Spectrum of Double Haploid Lines Derivedfrom IR64 and Wild Rice Species, Oryza rufipogonContained the Blast Resistance QTL (Pir). Dwinita W.Utami, A. Dinar Ambarwati, Aniversari Apriana, AtmitriSisharmini, Ida Hanarida, Didier Tharreau, and Santosa.This study was initiated to determine the spectrum resistanceof the candidate durable blast resistance variety containedthe QTL (quantitative trait locus), Pir1 and 2. This QTLwas mapped on chromosome 2 detected using the advancedbackcross population (BC5) from the wild rice speciesOryza rufipogon to IR64. Pir (1 and 2) also establishedon double haploid (DH) population derived from the selectedlines of BC2F3 population, progenies from the sameparents. The DH lines were developed to speed up the fixationprocess of the recessive alleles in the selected lines.Near isogenic lines with different blast resistance genes andcombination were used in this study comparing to the DHpopulation on their resistance spectrum using the knownavr gene isolates both on green house and field screening.The determination of the resistance spectrum will useful onthe prediction of durability of blast resistance gene in DHpopulation. The results of spectrum resistance test in greenhouse and field showed that Pir1and Pir2 segregated on 1 : 1proportion related with specific respond to blast avr genePH14 and CM28 resistance. Pir1 was identic to Pi33 or Pi25and Pir2 to Pitq5 on spectrum resistance
Parameter Genetik Jagung Populasi Bisma pada Pemupukan Berbeda. I. Ragam Aditif-Dominan Bobot Biji Jagung
Get PDFNew maize varieties could be obtained through improvement of their plant populations. The method used in selection in the crop improvement was based on values of their genetic parameters. Bisma is one of the maize varieties that has a broad genetic background. New maize varieties could be obtained by improving their population through selection under different environmental conditions. Genetic parameter value were estimated by conducting an experiment under NCD II crossing at Bogor. Twenty seven sets, which were developed from three females and three males of S1 as parents of each set, were evaluated under three different fertilization schemes. Results of the experiment showed that the additive genetic variance was significantly different from zero, and so among the different levels of fertilizer applications. The dominant variances was not significant under the three different levels of fertilization applications. The additive genetic variance was lower under the low level of fertilizer application than that on the higher level of fertilization application. This might be due to the scale effect. To reduce effect of scale, the data were transformed by dividing the grand mean value. After the data transformation, the genetic variance under the low level of fertilizer application tended to be greater than that under the higher level of fertilizer application. There was a tendency that population improvement of Bisma variety could be achieved better under lower level of fertilizer applications than under the higher ones
Genetic Mapping of SSR Markers in Eight Soybean Chromosomes Based on F2 Population B3462 x B3293
No full textGenetic Mapping of SSR Markers in Eight SoybeanChromosomes Based on F2 Population B3462 x B3293. IMade Tasma, Ahmad Warsun, Dani Satyawan, SaptowoJ. Pardal, and Slamet. Aluminum toxicity is one of the maincontrains for cultivating soybean in acid soils. GeneticHak Cipta © 2011, BB-Biogenmapping of SSR markers is one step for detecting aluminumtoxicitytolerant QTLs in soybean. Another step is tophenotype the same population at various aluminum-toxicityenvironments. The objectives of this study were to analyzethe segregation of SSR markers in progenies of an F2population and map the markers in 8 soybean chromosomes.The F2 population was previously developed bycrossing the Al-tolerant parent B3462 and the Al-sensitiveparent B3293. Polymorphic SSR markers in the parents wereused to PCR amplify DNA of the 100 F2 progenies. PCRproducts were separated using agarose or polyacrylamidegels. A Chi-Square test was done with a null hypothesis thatprogenies segregated in a 1 : 2 : 1 ratio. Results showed that125 SSR markers were polymorphics in the parents. Out of125 polymorphic markers, 122 were segregated in theprogenies of the F2 population. Among the segregatingmarkers, 114 were segregated in a 1 : 2 : 1 ratio. Only 8markers (5.6%) did not follow the 1 : 2 : 1 ratio. One hundredand nineteen SSR markers were mapped in 8 soybeanchromosomes. These include 18 markers in chromosomeA2, 10 in B1, 16 (C1), 16 (F), 10 (G), 23 (J), 16 (L), and 10 (N).Total genetic maps covered was 1,194.8 cM with averagemap distances between two adjacent markers of 10.7 cM.Further SSR marker enrichment is required to fill in the gapsof several chromosomal regions. Genetic maps presented inthis study should be useful for detection of Al-toxicitytolerant QTLs in soybean
Potensi Pemanfaatan Perangkat Diagnostik ELISA serta Variannya untuk Deteksi Patogen Tanaman
Get PDFDiseases aremajor constrains to agricultural crop productions in Indonesia.In the current free world trade system, the chances ofintroduction of plant quarantine agents are higher, and aredifficult to control, due to importation of seeds and otherplanting materials. Principles of the plant disease controlinclude exclusion and eradication. Early and accurate diseasediagnosis is an early and important step for a successfuldisease control. Enzyme-linked Immunosorbent Assay(ELISA) is a promising technique for an aneffective andefficient disease diagnosis. Some advantages of techniqueover the conventional and molecular diagnostic techniquesare economical use of reagents, high sensitivity, relativelysimple and quick, suitable for large numbers of samples,and adaptable for automation. In the past decade, severalvariants and kits of ELISA had been introduced, such asIndirect ELISA, F(ab’)2 ELISA, Dot Blot ELISA, and ImmunoFluorescence Assay (ELFA). Based on the solid membraneused, the Dot Blot ELISA some variants were developed,such as the NCM-ELISA, Tissue Blotting ELISA, dan Paper ELISA. The ELISA variants had different limit of detection levels. The limit detection of the variants for bacteria is ranging from 102-105 cells/ml, while those for viruses were from 1-10 ng/ml. The times required for the ELISA tests ranging from 5-48 hours. Models and components of ELISA kits for some viral and bacterial plant pathogens had been developed, but more are still needed since generally for each pathogen needs a different kit. The commercially available ELISA kits are limited in numbers, some of themare for pathogens that are not present in Indonesia. Production of ELISA kits for domestic uses will be more effective and efficent, particularly for pathogens that are present in the country. The ELISA kits are applicable not only fo detection and identification of pathogens, but also forecological study of the pathogens in conjuction with epidemiological study of the disease. This paper is a brief review on the ELISA technique and its variants and potential uses for detection of plant pathogen
Transformasi Genetik Kedelai dengan Gen Proteinase Inhibitor II Menggunakan Teknik Penembakan Partikel
Get PDFAn experiment was conducted at the Molecular Biology and Genetic Engineering Laboratory of BB-Biogen, Bogor with an objective to obtain transgenic soybean plants containing the proteinase inhibitor II (pinII) gene. The experiment consisted of three steps, i.e., optimalization of the soybean transformation technique using the gus gene; transformation of soybean using the pinII gene, and molecular analysis of the transformed soybean plants. Two type of explants (young embryo and cotyledon) were bombarded with pRQ6 plasmid containing the gus gene with the following treatment: Helium gas pressure (1100 psi and 1300 psi), shoot distance (5 and 7 cm), and number of bombardment (1x and 2x). The result of gus assay indicated that the best bombardment was done on young cotyledon explants with 1100 psi Helium pressure, shoot distance 5 cm, and 1x bombardment. Transformation of the soybean explant using the pinII gene (inside the pTWa plasmid) was conducted using the best bombardment treatment from the first activity. Two plants from c.v. Wilis (WP1, WP2) and three plants from c.v. Tidar (TP1, TP2, TP3) were recovered from regeneration and selection of the transformed explants. Molecular analysis of the regenerated plants using the PCR technique showed that only WP2 contained the pinII gene. This plant was fertile and will be used for further evaluation
Teknik Isolasi dan Kultur Protoplas Tanaman Padi
Get PDFProtoplastfusion or somatic hybridization technology is an alternativetechnology for production hybrids of plants that are difficultto be produced by conventional methods due to their sexualincompatibility. An experiment was conducted to developtechniques for isolation, purification, and culture of riceprotoplasts of cultivar IR64 and a wild rice species (Oryzaofficinalis). Optimization of protoplast isolation and purificationmethods from both rice genotypes were successfullydone. The highest protoplast density was obtained bydigesting embryonic callus or stems of young seedling in anenzyme solution containing of 2% cellulose, 0.1% pectolyase,0.5% macerozyme, 0.5% driselase, 5 mM ES, and 13% mannitolin CPW solution. The protoplast digestion was done forthree hours by soaking in the enzyme solution followed byshaking at 50 rpm under a room temperature. Purification ofthe protoplasts were done by separating them from plantdebris using a 25% sucrose solution. Protoplast regenerationwas not successful using although different media compositionsand conditions. Growth process from cell division tocell aggregate was only successful on IR64 protoplast cultureon a medium that contained AgNO3
Peningkatan Keragaman Genetik Tanaman melalui Keragaman Somaklonal
Get PDFHigh genetic variability’s are important factors in the development of new crop varieties. In vitro techniques are applicable for development of crop variability that is not found in the gene pool. One of the in vitro techniques that can be used for this purpose is the somaclonal variation technique. Somaclonal variation may be derived from genetic variations in explants and genetic variations in tissue cultures. Variations in the explant may be obtained from cell mutations or polysomic mutations of a certain tissue. Genetic variations in tissue culture may be caused by ploidy of chromosomes (endomitosis fusion), changes of chromosom structures (crossings), as well as changes of genes and cytoplasms. Changes of genetic characters may be improved if anorganic compound was added into the medium. To improve the plant tolerances to biotic or abiotic factors, selection components may also be added to the medium. Research results showed that somaclonal variation in tissue culture can improve genetic variations in plants. The variation produced in tissue culture provide chances to develop new plant genotipes. Many selection components, such as Gamma-ray irradiation, Al contents and low pH, pure toxin or filtrate, polyethylene glycol (PEG), and plant growth regulators can be used to improve somaclonal variations in many plants to produce new genotipes
Optimasi Sistem Regenerasi dan Transformasi Padi Varietas Elit Indonesia
Get PDFNew ricevariety can be generated by means of transgenic approach.Transgenic rice researches have been conducted in manyinstitutions worldwide using Japonica, Indica, and Javanicavarieties. The most cultivated rice in Indonesia is Indica.Indica type is known to have low responsive tissues inculture and transformation media when compared toJaponica rice. This research activity is aimed to optimizeregeneration and transformation systems of the Indonesianelite rice varieties, so that good method can be achieved tobe used in the generation of transgenic elite rice varieties ofIndica type. The research consisted of two activities:regeneration and transformation optimizations in varieties ofDodokan (upland rice) and Inpari 6 (irrigated rice).Immature embryo was used as the explant in this research.The optimization studies used 2 types of media, NBH (N6salts and vitamins, cassamino acid 0.5 g/l, L-proline 0.5 g/l,sucrose 20 g/l, D-glucose 10 g/l, 2.4-D 2 mg/l, NAA 1 mg/l, BA1 mg/l, agarose Type I 5.5 g/l) and NBH-M (N6 macro salts,B5 micro salts, and vitamins, 0.3 g/l cassamino acid, 3 g/l Lproline,20 g/l sucrose, 3 mg/l 2.4-D, 1 mg/l NAA, 1 mg/l BAP,5.5 agarose Type I), 2 types of regeneration media, R1 (MSbase media and vitamis, 0.3 g/l glutamine, 30 g/l sucrose, 2mg/l kinetin, 1 mg/l NAA, 3 g/l phytagel) and R2 (MS basemedia and vitamins, 2 g/l cassamino acid, 20 g/l sucrose, 30mg/l sorbitol, 2.5 mg/l kinetin, 0.25 mg/l NAA, 3 g/l phytagel).Optimization transformation of Indonesian elite rice varietiesused developed an empty plasmid pCAMBIA 1301 containinghpt gene. The transformation was conducted using twotypes of co-cultivation media, K1 (N6 macro salts, B5 microsalts, and vitamins, 0.5 g/l cassamino acid, 0.5 g/l L-proline,20 g/l sucrose, 10 g/l glucose, 2 mg/l 2.4-D, 1 mg/l NAA, 1mg/l BAP, 0.1 mM acetosyringone) and K2 (N6 macro salts,B5 micro salts, and vitamins, 0.5 g/l cassamino acid, 0.5 g/l Lproline,20 g/l sucrose, 10 g/l glucose, 2 mg/l 2.4-D, 1 mg/lNAA, 1 mg/l BAP, 0.2 mM acetosyringone). The resultsshowed that Inpari 6 could form embryonic calli in NBHmedia and further regenerated well in R1 media (13.8%).The co-cultivation media K1 generated more selected calliwhich then generated green plant of young embryocompared to K2. Inpari 6 showed higher regeneration ratesafter transformation (3.6%) compared to Dodokan (0%).Molecular analysis showed that all 11 transformants (Inpari6) tested contained the hpt gene. These results are expectedto support the development of transgenic Indica ricegeneration in Indonesia
Construction and Transformation of HVA1 Gene Expression Vector into Indonesian Elite Rice Varieties
Get PDFof drought in rice. HVA1 is one of the Late EmbryogenesisAbundant (LEA) protein group that plays a role on cellprotection during stresses. A study was done with anobjective to construct a plasmid vector expressing HVA1 andto transform it into Indonesian elite rice varieties. Materialsused in the study were plasmid pBY520 (source of HVA1;intermediate plasmid pRP9; plasmid pAY560326(backbone); restriction enzymes BamHI, HindIII, XhoI, andSpeI; T4 DNA ligase, and gel DNA extraction kit. Methodsused were standard procedure for plasmid vectorconstruction and molecular biology. Step I: the pBY520 andpRP9 were cut with BamHI and HindIII, and electrophoratedwith 1% agarose gel. DNA fragments of HVA1 and pRP9 werepurified, ligated with T4 DNA ligase, and transformed intoEscherichia coli DH5-α by heat shock. E. coli were grownonto solid medium (+ kanamycin 100 mg/l). A new plasmidDNA was isolated from single colony culture of the bacteria,confirmed, and named pRP9_HVA1. Step II: DNA ofpRP9_HVA1 and pAY560326 were cut with XhoI dan SpeIenzymes, purified, and ligated. The next procedure wassimilar to step I, and the resulted plasmid was confirmed byPCR and digestion with XhoI dan SpeI enzymes, and namedpAY_HVA1. Step III: pAY_HVA1 was first transformed intoAgrobacterium EHA-105 and then into rive varieties Ciherangand Inpari 6 using the early infection of scutellumtransformation method. Nine transgenic rice lines thatpositively contain HVA1 were obtained