scholarly journals Clustering Analysis and Genome Inference of Pisang Raja Local Cultivars (Musa spp.) from Java Island by Random Amplified Polymorphic DNA (RAPD) Marker

2019 ◽  
Vol 4 (2) ◽  
pp. 42 ◽  
Author(s):  
Rasyadan Taufiq Probojati ◽  
Didik Wahyudi ◽  
Lia Hapsari

Pisang Raja is an important local banana cultivar in the economy and cultural life in Indonesia, especially at Java. There are many Pisang Raja cultivars found on Java Island with various local names in each region, resulted in problems on taxonomic identification and grouping. Conventional research for grouping banana cultivars is still using morphological characters but considered inaccurate because of its subjectivity. This study aims to analyze the genetic diversity, grouping, and genome estimation of 13 local cultivars of Pisang Raja based on molecular approach using RAPD markers (OPA primers 1-20). Clustering and Principal Coordinates Analysis were performed to the amplified products using Paleontological Statistics (PAST) application version 3.15. Results showed that there were 12 primers which successfully amplified and produced DNA polymorphic bands in Pisang Raja, specifically OPA 1, OPA 2, OPA 3, OPA 4, OPA 5, OPA 8, OPA 16, OPA 17, OPA 18, OPA 19, and OPA 20. Pisang Raja cultivars considered have high genetic diversity, indicated by high polymorphic bands (95.17%) and low similarity coefficient values (0.2-0.6). Clustering and PCo analysis resulted in 3 clusters following its genomic group consist of AAA, AAB and ABB genomes, with Pisang Raja Bali as an outgroup (ABB). However, the separation of each cluster for genome inference was unclear. Cluster 1 consists of Pisang Raja Madu (AAB) and Raja Sereh (AAB). Cluster 2 consists of AAA and AAB genomes; includes Pisang Raja Jambe (AAA), Raja Kriyak (AAA), Raja Kutuk (AAB), Raja Brentel (AAB), Raja Seribu (AAB), and Raja Lini (AAB). Cluster 3 consists of AAA and AAB genomes, includes Pisang Raja Kisto (AAA), Raja Delima (AAA), Raja Bandung (AAB) and Raja Gareng (AAB). While Pisang Monyet (AAw) and Klutuk Wulung (BBw) as wild relatives were nested in Cluster 2. There were some different results of genome estimation based on RAPD markers compared to morphological characterization, and other molecular techniques. The use of RAPD markers is quite efficient and effective for studying genetic diversity and identifying genomes in bananas.

2019 ◽  
Vol 20 (3) ◽  
pp. 847-852
Author(s):  
TATI BARUS ◽  
RONALDO HALIM ◽  
ANASTASIA TATIK HARTANTI ◽  
PAULUS KEVIN SAPUTRA

Abstract. Barus T, Halim R, Hartanti AT, Saputra PK. 2019. Genetic diversity of Rhizopus microsporus from traditional inoculum of tempeh in Indonesia based on ITS sequences and RAPD marker. Biodiversitas 20: 847-852. The main microorganism for tempeh fermentation is Rhizopus microsporus. These days, many tempeh producers use commercial inoculum, such as ‘Raprima’ as resource of R. microsporus. As a result, the genetic diversity of R. microsporus that had been reported in Indonesia has diminished. Information about genetic diversity is needed as a basis to select R. microsporus as tempeh inoculum. This research aims to investigate the genetic diversity of R. microsporus from waru leaves based on Internal Transcribed Spacer (ITS) Sequence and Random Amplified Polymorphic DNA (RAPD) markers. A total of 25 R. microsporus were isolated from traditional inoculum waru leaves (Inoculum 1) and traditional inoculum other than waru leaves (Inoculum 2). Amplification of ITS sequence was done using universal primer pairs of ITS-4 and ITS-5. Amplification of RAPD markers was done using primers OPC-08, OPC-19, OPQ-6, R-108, OPA-09 and OPJ-20. ITS sequence was not sufficient to compare the similarities among R. microsporus. On the other hand, RAPD markers successfully compared the similarities among 25 R. microsporus. A total of 25 R. microsporus were divided into 9 clusters. R. microsporus from Inoculum 1 grouped into Cluster 1, Cluster 3 and Cluster 4-8. Inoculum 2 grouped into Cluster 2 and Cluster 9. R. microsporus from tempeh grouped into Cluster 4 and was different from Inoculum 1 and Inoculum 2, except for TB3.


2013 ◽  
Vol 22 (2) ◽  
pp. 127-136
Author(s):  
MM Uddin ◽  
MI Khalil ◽  
MS Haque ◽  
MB Meah

Random amplified polymorphic DNA (RAPD) assay was performed to estimate genetic polymorphisim in ten chili cultivars. Out of 12 primers four (OPA11, OPB03, OPB04 and OPB17) showed amplification of genomic DNA and generated 21 distinct score able bands of which 17 (80.95%) were polymorphic. The highest percentage (85.71) polymorphic locus was found in OPB03 while the lowest (66.67) in OPA11. The highest genetic distance was computed between Jamalpur Balujuri and Matal marich with the lowest genetic identity as against the lowest genetic distance between Hajari marich and Balujuri marich. The UPGMA dendogram indicated segregation of ten chili varieties and genotypes into two main clusters. Variety Bogra marich and Matal marich formed cluster 1 and Balujuri marich, Deshi marich, Jamalpuri balujuri, Bindu marich, Syloti, Hajari, Biroli city, and the genotype Ausadhebrara grouped in cluster 2. The result indicates the genetic diversity among the chili cultivars and RAPD marker could be used for improvement of chili varieties. DOI: http://dx.doi.org/10.3329/ptcb.v22i2.14201 Plant Tissue Cult. & Biotech. 22(2): 127-136, 2012 (December)


2019 ◽  
Vol 63 (1) ◽  
pp. 15-24
Author(s):  
Soumen Saha ◽  
Tarak Nath Dhar ◽  
Parthadeb Ghosh ◽  
Tulsi Dey

The aim of this research was to assess the genetic diversity of sesame (Sesamum indicum L.) and also to reveal the genetic relationships using the Random Amplified Polymorphic DNA (RAPD) markers. Fifteen sesame germplasms were collected from seven districts or four zones of West Bengal, India. A high genetic diversity was revealed by ten RAPD primers within and among the fifteen germplasms. The value of Jaccard’s similarity coefficients among and within the fifteen germplasms ranged from 0.287 to 0.725 which indicated high degree of genetic variability. Cluster analysis using Unweighted Pair Group Method with Arithmetic Mean (UPGMA) grouped all the germplasms into three main clusters. Analysis of various genetic diversity indices strongly indicated high level of genetic diversity among the populations of four different regions. UPGMA analysis of four populations resulted into two groups and the results of Principal Coordinates Analysis (PCoA) depicted a clear distinction among the germplasms.


2010 ◽  
Vol 90 (4) ◽  
pp. 443-452 ◽  
Author(s):  
T. Karuppanapandian ◽  
H W Wang ◽  
T. Karuppudurai ◽  
J. Rajendhran ◽  
M. Kwon ◽  
...  

The DNA fingerprinting methodologies, random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR), were used to estimate genetic diversity and relationships among 20 black gram (Vigna mungo L. Hepper) varieties. Thirty selected RAPD primers amplified 255 bands, 168 of which were polymorphic (66.5%). On average, these primers produced 8.5 bands, 5.6 of which were polymorphic. Polymorphic band number varied from 2 (A-05) to 10 (OPA-02), with sizes ranging from 100 to 2550 bp. Twenty-four selected ISSR primers produced 238 amplified products, 184 of which were polymorphic (77.8%). On average, these primers generated 9.8 bands, with 7.7 polymorphic bands ranging in number from 4 (ISSR-13) to 11 (ISSR-03), and size from 100-2650 bp. Genetic relationships were estimated using similarity coefficient (Jaccard’s) values between different accession pairs; these varied from 30.7 to 85.0 for RAPD, and from 37.2 to 88.4 with ISSR. UPGMA analysis indicated that the varieties ranged in similarity from 0.50 to 1.00 (mean of 0.75) for RAPD, and from 0.47 to 1.00 (mean of 0.76) with ISSR. Cluster analysis of RAPD and ISSR results identified three clusters with significant bootstrap values, which revealed greater homology between the varieties. Principal coordinates analysis also supported this conclusion. Among the black gram varieties, WBU-108 and RBU-38 were highly divergent, whereas LBG-648 and LBG-623 were genetically similar. The markers generated by RAPD and ISSR assays can provide practical information for the management of genetic resources and these results will also provide useful information for the molecular classification and breeding of new black gram varieties.Key words: Black gram, cluster analysis, genetic diversity, ISSR, molecular markers, RAPD


Genome ◽  
1994 ◽  
Vol 37 (6) ◽  
pp. 1011-1017 ◽  
Author(s):  
Zhao-Wei Liu ◽  
Robert L. Jarret ◽  
Ronny R. Duncan ◽  
Stephen Kresovich

Random amplified polymorphic DNA (RAPD) markers were used to assess genetic relationships and variation among ecotypes of the turfgrass seashore paspalum (Paspalum vaginatum Swartz). Vegetative tissues or seeds of 46 seashore paspalum ecotypes were obtained from various locations in the United States, Argentina, and South Africa. Leaf DNA extracts were screened for RAPD markers using 34 10-mer random primers. A total of 195 reproducible RAPD fragments were observed, with an average of six fragments per primer. One hundred and sixty-nine fragments (87% of the total observed) were polymorphic, among which 27 fragments (16%) were present in three or less ecotypes, indicating the occurrence of a high level of genetic variation among the examined accessions of this species. Cluster analysis (UPGMA) and principal coordinates analysis were performed on the RAPD data set. The results illustrate genetic relationships among the 46 ecotypes, and between ecotypes and their geographical origins. Ecotypes from southern Africa could be differentiated from the U.S. and most of the Argentinean ecotypes. With a few exceptions, ecotypes collected from Argentina, Hawaii, Florida, and Texas were separated into distinct clusters.Key words: RAPDs, polymerase chain reaction, genetic diversity, phenetic analysis.


Genetika ◽  
2014 ◽  
Vol 46 (2) ◽  
pp. 331-342 ◽  
Author(s):  
Dragana Miladinovic ◽  
Ksenija Taski-Ajdukovic ◽  
Nevena Nagl ◽  
Branislav Kovacevic ◽  
Aleksandra Dimitrijevic ◽  
...  

Random amplified polymorphic DNA (RAPD) markers were used to detect polymorphism among accessions of wild sunflower species H?lianthus maximiliani, Helianthus tuberosus, Helianthus mollis and Helianthus rigidus with different tolerance to mid-stalk white rot and selection of potential markers for different levels of tolerance to this disease. Estimates of genetic variation showed that genetic diversity was equally distributed between Helianthus species and within them. Cluster analysis corresponded to the phylogenetic relations within the genus Helianthus. The results obtained by principal coordinates analysis (PCoA), where the first two principal coordinates accounted for 83.7% of total variation, perfectly coincided with the results of cluster analysis. Contingency coefficient significance test showed that most of the used primers generated bands associated with some level of tolerance or susceptibility to mid- stalk white rot. Furthermore, contingency analysis showed that primer C12 generated bands associated with resistance (100%) to mid-stalk white rot both in H. mollis and in all accessions, while primer X18 generated bands significantly associated with high tolerance (75%) in H. rigidus, H. mollis as well as in all tested accessions. The C15-600 bp locus was found to be significantly associated with high tolerance (75%) in all accessions, and medium tolerance (50%) in H. mollis.


2014 ◽  
Vol 1 (1) ◽  
pp. 1 ◽  
Author(s):  
Budi Martono ◽  
Laba Udarno

<p>Informasi keragaman genetik dan ketersediaan plasma nutfah teh (Camellia sinensis) diperlukan dalam perakitan varietas unggul. Keragaman genetik berdasarkan penanda DNA dapat memberikan hasil yang lebih konsisten karena tidak dipengaruhi lingkungan. Dalam penelitian ini sebanyak 9 genotipe teh dianalisis keragamannya menggunakan enam penanda RAPD (OPA 03, OPA 05, OPB 04, OPB 06, OPC 06, dan OPD 08). Penelitian dilakukan mulai bulan Maret sampai Mei 2013 di Laboratorium Terpadu Biotrop Bogor. Perhitungan koefisien kesamaan genetik dan analisis gerombol dilakukan dengan menggunakan perangkat lunak NTSYSpc versi 2.02. Sebanyak 54 lokus penanda RAPD berhasil diamplifikasi menggunakan enam primer dan 47 lokus di antaranya memiliki alel yang polimorfik (87,04%). Hasil analisis gerombol berdasarkan kesamaan genetiknya mengelompokkan 9 genotipe ke dalam enam kelompok. Empat kelompok (I, II, IV, V) masing-masing terdiri atas satu genotipe, sementara dua kelompok yang lain yaitu kelompok III dan VI masing-masing beranggotakan tiga dan dua genotipe.</p><p>Kata Kunci: Camellia sinensis, diversitas genetik, penanda RAPD</p><p>The availability of diverse tea (Camellia sinensis) germplasms as well as the information about their genetic diversity is required for plant breeding program. Genetic diversity analysis based on DNA marker is known to be more effective since the markers provide more consistent results. In this study, nine tea genotypes were evaluated for their genetic diversity using six Random Amplified Polymorphic DNA (RAPD) markers (OPA 03, OPA 05, OPB 04, OPB 06, OPC 06, and OPD 08). The study was conducted from March to May 2013 in the Integrated Laboratory of Biotrop Bogor. The estimation of genetic similarity and the cluster analysis were done using NTSYSpc version 2.02. Of the six RAPD markers used in this study, a total of 54 RAPD marker loci have been successfully amplified. In which, 47 loci (87.04%) were polymorphic and subsequently used for the evaluation of tea genotypes. The results of cluster analysis showed that those tea genotypes were clustered into six groups. Each of four groups (I, II, IV, V) consisted of only one genotype. Meanwhile, the other two groups (III and VI) had three and two genotypes, respectively.</p>


2013 ◽  
Vol 6 (1-2) ◽  
pp. 51-63
Author(s):  
SM Faisal ◽  
MS Haque ◽  
KM Nasiruddin ◽  
MM Islam ◽  
MA Shrafuzzaman ◽  
...  

Genetic variability among the genotypes of any species could be utilized for its improvement. PCR-based Random Amplified Polymorphic DNA (RAPD) technique was used to determine the genetic diversity and relationship among 10 cucumber varieties and genotypes. Five decamer primers were used to amplify genomic DNA and the primers yielded a total of 54 bands of which 36 bands were polymorphic and 18 bands were monomorphic. The UPGMA dendrogram based on Nei’s (1972) genetic distance indicated segregation of 10 cucumber varieties and genotypes into two main clusters. Variety Joti alone grouped in cluster 1 while variety Green Master, Shahi-50, Shikha, Shila, Shital, Naogaon-5, Shohag-50, Giant Long and genotype CS-043 grouped in cluster 2. Variety Shila was very close to variety Shital with the least genetic distance (0.1712). The highest genetic distance (0.5352) was found between Joti and Naogaon-5. DOI: http://dx.doi.org/10.3329/cujbs.v6i1-2.17081 The Chittagong Univ. J. B. Sci.,Vol. 6(1&2):51-63, 2011


2019 ◽  
Vol 61 (4) ◽  
pp. 255-266
Author(s):  
◽  
Rini Hafzari ◽  
Tia Setiawati ◽  
Budi Irawan ◽  
Joko Kusmoro

Abstract Conservation of bamboos for future exploitation as fuel, fibre and as an ingredient for cosmetics depends on knowledge of its natural genetic variation. The study of molecular genetic diversity in bamboos will provide important information for its conservation. This article reports on the genetic diversity in 25 species representing five genera of bamboos found in Indonesia using Random Amplified Polymorphic DNA (RAPD) molecular markers. Out of 40 primers, 24 primers produced 1107 total bands and 86.21% of polymorphic bands across the 25 species. Sixteen bands were uniquely found in one species only and their presence or absence helped to define nine bamboo species. RAPD band sizes ranged from 162 to 2247 base pairs. A dendrogram based on the similarity coefficient of Dice divided the bamboo species into three big clusters. In conclusion, RAPD can capture the diversity among five different bamboo genera and has a great potential to be used in the study of genetic diversity in Indonesian bamboos.


2003 ◽  
Vol 128 (5) ◽  
pp. 741-746 ◽  
Author(s):  
N. Nikoloudakis ◽  
G. Banilas ◽  
F. Gazis ◽  
P. Hatzopoulos ◽  
J. Metzidakis

Random amplified polymorphic DNA (RAPD) markers were used to study the genetic diversity and to discriminate among 33 Greek olive (Olea europaea L.) cultivars. Three feral forms from Crete and five foreign cultivars recently introduced into Greece were also included. Nineteen primers were selected which produced 64 reproducible polymorphic bands in the 41 olive genotypes studied, with an average of 3.4 informative markers per primer. The RAPD markers resulted in 135 distinct electrophoretic patterns, with an average of 7.1 patterns per primer. Based on either unique or combined patterns, all genotypes could be identified. Genetic similarities between genotypes were estimated using the Dice similarity index and these indicated that a high degree of diversity exists within the Greek olive germplasm. Using the unweighted pair-group method (UPGMA) most cultivars were clustered into two main groups according to their fruit size or commercial use (table or olive oil). However, poor correlation was detected between clustering of cultivars and their principal area of cultivation. RAPD marker data were subjected to nonmetric multidimentional scaling (NMDS) which produced results similar to those of the UPGMA analysis. The results presented here contribute to a comprehensive understanding of cultivated Greek olive germplasm and provide information that could be important for cultural purposes and breeding programs.


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