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Phylogenetic and Maturity Analyses of Sixty Soybean Genotypes Used for DNA Marker Development of Early Maturity Quantitative Trait Loci in Soybean

Jurnal AgroBiogen Vol 7, No 1 (2011): Jurnal AgroBiogen
Publisher : Jurnal AgroBiogen

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Abstract

The Indonesian soybean productivity is still very low with the national average of 1.3 t/ha. One means to improve national soybean productivity is by manipulating harvest index by cultivating very early maturing soybean cultivars. Development of early maturing soybean cultivars can be expedited by using marker-aided selection. The objective of this study was to select parental lines having contrasted maturity traits and selected parents must be genetically distance. The parents then were used to develop F2 populations for detecting early maturity QTL in soybean. Maturity tests of 60 soybean genotypes were conducted at two locations, Cikeumeuh (Bogor) and Pacet (Cianjur) using a randomized block design with three replications. Genomic DNA of the 60 genotypes were analyzed using 18 SSR markers and genetic relationship was constructed using the Unweighted Pair-Group Method Arithmatic through Numerical Taxonomy and Multivariate System program version 2.1-pc. Results showed that the 60 genotypes demonstrated normal distribution in both locations for days to R1 (32-48d), days to R3 (35-55d), days to R7 (75-92d), and days to R8 (78-99d). Four early maturing genotypes and three late genotypes were obtained. Total SSR alleles observed were 237 with average allele per locus of 12.6 (3-29), and average PIC value of 0.78 (0.55-0.89). Genetic similarity among genotypes ranges from 74.8-95%. At similarity level 77% divided the genotypes into six clusters (the four selected early maturing genotypes located in clusters III and IV, while the three late genotypes located in cluster II). Based on maturity data, pubescent color, and phygenetic analysis seven parents were selected (four early maturing genotypes B1430, B2973, B3611, B4433 and three late genotypes B1635, B1658, and B3570). Twelve F2 populations were developed with the aid of SSR markers Satt300 dan Satt516. Two of the populations will be used to develop DNA markers for earliness in soybean.

Konstruksi Pustaka Genom Kakao (Theobroma cacao L.) untuk Sekuensing Genom Total Menggunakan Next Generation Sequencing HiSeq2000

Jurnal Tanaman Industri dan Penyegar Vol 3, No 2 (2012): Buletin Riset Tanaman Rempah Dan Aneka Tanaman Industri
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

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Abstract

Pemuliaan kakao secara konvensional memerlukan waktu panjang (10-15 tahun). Pemanfaatan marka DNA akan memperpendek siklus pemuliaan kakao. Tujuan penelitian ini adalah mengkonstruksi pustaka genom tiga genotipe kakao yang dapat digunakan untuk sekuensing genom total kakao menggunakan NGS HiSeq2000 dan mendapatkan data resekuen genom total tiga genotipe kakao.  Bahan tanaman terdiri dari tiga klon unggul kakao (ICCR02, ICCR04, dan SUL02) diperoleh dari Balittri, Pakuwon.  DNA genomik diisolasi dari daun muda sebagai bahan konstruksi pustaka genom total. Sekuensing pustaka dilakukan pada mesin HiSeq2000 mengikuti protokol dari Illumina. Pustaka genom yang telah berhasil dikonstruksi berukuran 300 pasang basa (bp) masing-masing dengan konsentrasi 14,70 ng/µL (ICCR02), 15,20 ng/µL (ICCR04), dan 12,90 ng/µL (SUL02). Ukuran dan konsentrasi pustaka genom yang dihasilkan sangat ideal untuk sekuensing menggunakan HiSeq2000. Sekuensing ketiga genom menghasilkan data sekuen 52,9 x 109 bp.  Klaster DNA pustaka genom memiliki nilai Q scores>30 (75,0%) dengan tingkat kesalahan pembacaan basa rendah (1,47%).  Nilai densitas klaster, persen klaster PF, intensitas basa, persen phasing, dan persen prephasing menunjukkan kualitas klaster pustaka genom ketiga genotipe kakao termasuk kategori pustaka ideal. Data sekuen yang dihasilkan juga sangat ideal untuk identifikasi marka SNP genom kakao. Koleksi marka SNP digunakan untuk identifikasi gen pengendali karakter penting kakao dan pemuliaan berbasis marka DNA untuk memperpendek siklus pemuliaan kakao. Genomic Library Construction Of Cocoa (Theobroma Cacao L.) For Whole Genome Sequensing Using A Next Generation Sequencer Hiseq2000Conventional cocoa breeding is slow and takes about 10-15 years to complete a breeding cycle. Applying genomic technology using DNA markers will significantly decrease cocoa breeding cycle. The objectives of this study were to construct cocoa whole genome genomic libraries to be used for resequencing the whole genome of cocoa and obtain whole genome resequence data of three cocoa genotypes. Three Indonesian cocoa genotypes (ICCR02, ICRR04, and SUL02) were used. DNA genomic was isolated from young leaf and used to construct genomic DNA libraries and generate DNA clusters. DNA clusters were sequenced using a HiSeq2000 platform. The whole genome libraries of the cocoa genotypes were successfully constructed. The library size was 300 bp with concentrations of 14.70 ng/µL (ICCR02), 15.20 ng/µL (ICCR04), and 12.90 ng/µL (SUL02), respectively. The genomic library size and concentrations are suitable for sequencing study using the NGS HiSeq2000. Total sequencing output obtained was 52.9 x 109 bp. The genomic library clusters resulted during the sequencing process demonstrated the Q scores > 30 of 75.0% with low error sequencing rate of 1.47%. Cluster densities, percentage of cluster PF, base intensity, and percentage of phasing and prephasing indicated the cluster quality of the genomic libraries is classified as an ideal one to be used for resequencing study using NGS HiSeq2000. The resequence data were ideal for SNP marker discovery. SNP markers are used to identify economically important genes of cocoa and marker-aided cocoa breeding to decrase the cocoa breeding cycle.

Konstruksi Pustaka Genom Kakao (Theobroma cacao L.) untuk Sekuensing Genom Total Menggunakan Next Generation Sequencing HiSeq2000

Jurnal Tanaman Industri dan Penyegar Vol 3, No 2 (2012): Buletin Riset Tanaman Rempah Dan Aneka Tanaman Industri
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

Show Abstract | Original Source | Check in Google Scholar | Full PDF (368.862 KB)

Abstract

Pemuliaan kakao secara konvensional memerlukan waktu panjang (10-15 tahun). Pemanfaatan marka DNA akan memperpendek siklus pemuliaan kakao. Tujuan penelitian ini adalah mengkonstruksi pustaka genom tiga genotipe kakao yang dapat digunakan untuk sekuensing genom total kakao menggunakan NGS HiSeq2000 dan mendapatkan data resekuen genom total tiga genotipe kakao.  Bahan tanaman terdiri dari tiga klon unggul kakao (ICCR02, ICCR04, dan SUL02) diperoleh dari Balittri, Pakuwon.  DNA genomik diisolasi dari daun muda sebagai bahan konstruksi pustaka genom total. Sekuensing pustaka dilakukan pada mesin HiSeq2000 mengikuti protokol dari Illumina. Pustaka genom yang telah berhasil dikonstruksi berukuran 300 pasang basa (bp) masing-masing dengan konsentrasi 14,70 ng/µL (ICCR02), 15,20 ng/µL (ICCR04), dan 12,90 ng/µL (SUL02). Ukuran dan konsentrasi pustaka genom yang dihasilkan sangat ideal untuk sekuensing menggunakan HiSeq2000. Sekuensing ketiga genom menghasilkan data sekuen 52,9 x 109 bp.  Klaster DNA pustaka genom memiliki nilai Q scores>30 (75,0%) dengan tingkat kesalahan pembacaan basa rendah (1,47%).  Nilai densitas klaster, persen klaster PF, intensitas basa, persen phasing, dan persen prephasing menunjukkan kualitas klaster pustaka genom ketiga genotipe kakao termasuk kategori pustaka ideal. Data sekuen yang dihasilkan juga sangat ideal untuk identifikasi marka SNP genom kakao. Koleksi marka SNP digunakan untuk identifikasi gen pengendali karakter penting kakao dan pemuliaan berbasis marka DNA untuk memperpendek siklus pemuliaan kakao. Genomic Library Construction Of Cocoa (Theobroma Cacao L.) For Whole Genome Sequensing Using A Next Generation Sequencer Hiseq2000Conventional cocoa breeding is slow and takes about 10-15 years to complete a breeding cycle. Applying genomic technology using DNA markers will significantly decrease cocoa breeding cycle. The objectives of this study were to construct cocoa whole genome genomic libraries to be used for resequencing the whole genome of cocoa and obtain whole genome resequence data of three cocoa genotypes. Three Indonesian cocoa genotypes (ICCR02, ICRR04, and SUL02) were used. DNA genomic was isolated from young leaf and used to construct genomic DNA libraries and generate DNA clusters. DNA clusters were sequenced using a HiSeq2000 platform. The whole genome libraries of the cocoa genotypes were successfully constructed. The library size was 300 bp with concentrations of 14.70 ng/µL (ICCR02), 15.20 ng/µL (ICCR04), and 12.90 ng/µL (SUL02), respectively. The genomic library size and concentrations are suitable for sequencing study using the NGS HiSeq2000. Total sequencing output obtained was 52.9 x 109 bp. The genomic library clusters resulted during the sequencing process demonstrated the Q scores > 30 of 75.0% with low error sequencing rate of 1.47%. Cluster densities, percentage of cluster PF, base intensity, and percentage of phasing and prephasing indicated the cluster quality of the genomic libraries is classified as an ideal one to be used for resequencing study using NGS HiSeq2000. The resequence data were ideal for SNP marker discovery. SNP markers are used to identify economically important genes of cocoa and marker-aided cocoa breeding to decrase the cocoa breeding cycle.

Genetic Mapping of SSR Markers in Eight Soybean Chromosomes Based on F2 Population B3462 x B3293

Jurnal AgroBiogen Vol 7, No 2 (2011): Oktober
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

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Abstract

Genetic 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.

Phylogenetic and Maturity Analyses of Sixty Soybean Genotypes Used for DNA Marker Development of Early Maturity Quantitative Trait Loci in Soybean

Jurnal AgroBiogen Vol 7, No 1 (2011): April
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Original Source | Check in Google Scholar | Full PDF (183.856 KB)

Abstract

Phylogenetic and Maturity Analyses of Sixty SoybeanGenotypes Used for DNA Marker Development of EarlyMaturity Quantitative Trait Loci in Soybean. I MadeTasma, Dani Satyawan, Ahmad Warsun, MuhamadYunus, and Budi Santosa. The Indonesian soybeanproductivity is still very low with the national average of 1.3t/ha. One means to improve national soybean productivity isby manipulating harvest index by cultivating very earlymaturing soybean cultivars. Development of early maturingsoybean cultivars can be expedited by using marker-aidedselection. The objective of this study was to select parentallines having contrasted maturity traits and selected parentsmust be genetically distance. The parents then were used todevelop F2 populations for detecting early maturity QTL insoybean. Maturity tests of 60 soybean genotypes wereconducted at two locations, Cikeumeuh (Bogor) and Pacet(Cianjur) using a randomized block design with threereplications. Genomic DNA of the 60 genotypes wereanalyzed using 18 SSR markers and genetic relationship wasconstructed using the Unweighted Pair-Group MethodArithmatic through Numerical Taxonomy and MultivariateSystem program version 2.1-pc. Results showed that the 60genotypes demonstrated normal distribution in bothlocations for days to R1 (32-48d), days to R3 (35-55d), days toR7 (75-92d), and days to R8 (78-99d). Four early maturinggenotypes and three late genotypes were obtained. TotalSSR alleles observed were 237 with average allele per locusof 12.6 (3-29), and average PIC value of 0.78 (0.55-0.89).Genetic similarity among genotypes ranges from 74.8-95%.At similarity level 77% divided the genotypes into six clusters(the four selected early maturing genotypes located inclusters III and IV, while the three late genotypes located incluster II). Based on maturity data, pubescent color, andphygenetic analysis seven parents were selected (four earlymaturing genotypes B1430, B2973, B3611, B4433 and threelate genotypes B1635, B1658, and B3570). Twelve F2populations were developed with the aid of SSR markersSatt300 dan Satt516. Two of the populations will be used todevelop DNA markers for earliness in soybean.

Genetic Diversity Analysis of Jatropha Curcas Provenances Using Randomly Amplified Polymorphic DNA Markers

Jurnal AgroBiogen Vol 7, No 1 (2011): April
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

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Abstract

Genetic Diversity Analysis of Jatropha CurcasProvenances Using Randomly Amplified PolymorphicDNA Markers. Dani Satyawan and I Made Tasma.Jatropha curcas nuts are rich in oil that is higly suitable forHak Cipta © 2011, BB-Biogenthe production of bio-diesel or to be used directly inmodified diesel engines. The objective of this study was toassess the extent of genetic diversity among 50 J. curcasprovenances and one accession of J. integerrima usingRAPD markers. The fifty J. curcas provenances werecollected from ecologically diverse regions of Indonesia, andplanted in the Pakuwon Experimental Station (Sukabumi,West Java). Fourteen RAPD primers with 60-80% G+Ccontent were used in this genetic diversity analysis andproduced 64 bands with 95.7% polymorphism level. ThePolymerase Chain Reactions used to generate the RAPDbands sometimes produced inconsistent and nonreproducibleresults, necessitating the duplication of eachreaction to prevent scoring errors. Sixty one validated bandswere subsequently used for genetic diversity analysis usingUnweighted Pair Group Method Arithmetic (UPGMA)method and Dice coefficients. It was shown that thesimilarity coefficients among the provenances ranged from0.2 to 0.98 with an average similarity of 0.75. Dendrogramanalysis produced two major groups of provenances, withone outlier from South Lampung. There was no tendency forprovenances originated from nearby regions to clustertogether in each group, and several provenances showedmore similarities with provenances originated from distantregions. This pattern lent credence to reports that Jatrophawas introduced to Indonesia around four centuries ago andwas mainly spread by humans. Based on the meansimilarities among the accessions and their clusteringpattern, the genetic diversity of the Jatropha collectionappeared to be fairly low. Future additions of geneticmaterials from more diverse genetic background will benecessary to maintain the current progress of Jatrophaimprovement program.

Analisis Kekerabatan 50 Aksesi Kelapa Sawit (Elaeis guineensis Jacq.) Asal Kamerun Berdasarkan Marka Mikrosatelit

Jurnal AgroBiogen Vol 9, No 1 (2013): April
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

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Abstract

Genetic diversity of theIndonesian oil palm collection remains low and collectionenrichness through exploration activities from the center oforigins is required. In 2009, 103 oil palm accessions fromCameroon were collected at the National Oil Palm GeneticResources Collection located at the District of Sijunjung,West Sumatera. The objectives of the present study were toanalayze the 50 Cameroon-originated oil palm accessions inorder to: (1) determine polymorphism levels of the SSRmarkers used; (2) understand diversity levels of the oil palmaccessions tested; and (3) analyze accessions potentiallyused for germ plasm collection. Fifty oil palm accessionswere used in this study. DNA was isolated from leaves of theselected individual plants representing each of theaccessions. DNA was analyzed with 12 SSR markers. Adendrogram was constructed using the UPGMA throughNumerical Taxonomy and Multivariate System programversion 2.1-pc. Results showed that SSR markers useddemonstrated the average number of alleles per locus of 3.6(2-6). The polymorphism level was 0.53 (0.21-0.73). Thephylogenetic analysis resulted nine clusters with geneticdiversity between two accessions ranged from 4-82%. Tenaccessions (20%) showed low genetic diversity (<10%) butthe accessions demonstrated high diversity in floweringtime. Eleven accessions showed medium diversity level (27-42%). Five accessions demonstrated high genetic diversitylevel (45-82%). A confirmation study using more SSRmarkers is recommended. This study finding may be usefulin planning the oil palm germ plasm collection activities.

Genetic Diversity Analysis and F2 Population Development for Breeding of Long Juvenile Trait in Soybean

Jurnal AgroBiogen Vol 14, No 1 (2018): June
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

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Abstract

Genetic diversity analysis using molecular markers is an important step for selecting appropriate parents in a soybean breeding program. The aims of this study were to (1) analyze genetic diversity of 29 soybean genotypes assessed with 27 SSR markers for selecting appropriate parents and (2) develop F2 populations to be used for breeding long juvenile (LJ) trait in soybean tobe cultivated in short photoperiod condition. The soybean genotypes used consisted of 11 Indonesian soybean genotypes and 18 genotypes introduced from the USA. F2 populations were developed by crossing Grobogan with three introduced genotypes carrying LJ character. The PIC values of the 27 SSR markers ranged from 0.87 to 0.96. Cluster analysis resulted in three mainclusters at coefficient similarity of 0.76. The five LJ introduced accessions and the nine Indonesian genotypes showed high genetic distances and are useful as parent pairs for developing breeding populations. The F1 progeny phenotypicperformances of the cross far exceeded the performaces of both parents. Three F2 populations were developed by crossing the distantly related soybean genotypes. The F2 populations were verified by using SSR markers and it was found that they segregated in a 1:2:1 ratio confirming the segregation ratio of codominant SSR markers. The F2 populations should be useful for breeding LJ characters to improve soybean productivity in low latitude tropical countries such as Indonesia, which has day length of approximately 12 h all year round.

Mapping of Resistance Genes to Brown Planthopper in Untup Rajab, an Indonesian Local Rice Variety

Jurnal AgroBiogen Vol 14, No 2 (2018): December
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

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Abstract

Brown planthopper (BPH) is a major rice pest in Indonesia. The most economical and effective approach to control the insect pest is by using resistant varieties. Exploring for resistance genes is, therefore, a prerequisite for effective breeding program for BPH resistance. This study aimed to map BPH resistance genes in Untup Rajab, an Indonesian local rice variety. Genetic map was constructed using an F2 population from a cross between TN-1 and Untup Rajab, and SNP markers from RiceLD SNP Chip. Phenotyping was performed using bulk seedling test on F2:3 seedlings against two BPH populations, i.e. X1 and S1. Four QTLswere identified on chromosomes 5, 6, 8, and 11 with PVE values of 7.63%, 9.40%, 17.66%, and 3.05%, respectively. Relatively normal distribution of resistance phenotype and the relatively low PVE values indicate that Untup Rajab has a quantitative resistance to BPH with two different resistance loci identified for each BPH test population. The QTL on chromosome 8 overlaps with OsHI-LOX gene, which is associated with resistance to BPH, and adjacent to another QTL for resistance to green leafhopper. The QTL on chromosome 6 was found near OsPLDα4 and OsPLDα5 genes which are related to BPH resistance. Meanwhile, the QTL intervals on chromosome 5 and 11 did not overlap with any known BPH QTLs or genes, which make them attractive candidates for novel BPH resistance gene discovery.