• Journal of Life Science
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• v.13 no.5
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• pp.590-595
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• 2003

The objective of this study was to assess of genetic variation within and between pumpkin species including Cucurbita maxima, C. moschata, C. pepo and C. maxima${\times}$C. moschata using RAPD markers. The 16 primers showed the amplification of 136 scorable fragments ranging from about 100 bp to 2300 bp. A total of 94 DNA fragments were polymorphic with an average 5.9 polymorphic bands per primer. A species $(C. maxima\timesC. moschata)$ has the highest number of polymorphic loci. Based on obtained data, UPGMA cluster analysis was conducted. Twenty pumpkin cultivars were classified into three large categories and identified genetic distance of cluster ranging from 0.38 and 1.00. Clustering was in accordance with the division of Curcurbitaceae into four species, C. maxima, C. moschata, C. pepo and C. $C. maxima\timesC. moschata$. Therefore, RAPD method may be essential tool for enabling discrimination of pumpkin cultivars.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.11
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• pp.1556-1560
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• 2006

Six breeds of riverine buffalo viz. Murrah, Mehsana, Jaffrabadi, Nagpuri, Nili-Ravi and Bhadawari were characterized using FAO-recommended cattle specific microsatellite markers. Among the total of twenty microsatellite markers screened to explore genomic variability of six buffalo breeds, only ten were polymorphic in nature. Four out of ten polymorphic microsatellite loci were rated as informative. The numbers of alleles detected ranged from 2 to 7, with a mean of $5.5{\pm}0.07$ per microsatellite marker. The most polymorphic marker was BM1818 with a total of 7 alleles present at this locus. One breed specific marker was found in each of Mehsana (BM1818) and Bhadawari (ILSTS030) and four were found in Jaffarabadi (BM1818, ILSTS030, ILSTS054 and ILSTS011). Genetic distance (Ds) between the Mehsana and Bhadawari breed was the maximum (0.29), followed by Murrah and Mehsana (0.27), and Nili-Ravi and Bhadawari (0.26). The lowest Ds was found between the Jaffrabadi and Nagpuri breeds which was only 0.05. The highest divergence time of 1318 years was established between Mehsana and Bhadawari breeds whereas it was found to be lowest (272 years) between the Jaffrabadi and Nagpuri breeds.

• Mycobiology
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• v.47 no.4
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• pp.466-472
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• 2019

For the purpose of protecting the rights of Lentinula edodes breeders, we developed a new simple sequence repeat (SSR) marker set consisting only of genetically independent tetranucleotide or longer core motifs. Using available genome sequences for five L. edodes strains, we designed primers for 13 SSR markers that amplified polymorphic sequences in 20 L. edodes cultivars. We evaluated the independence of every possible marker pair based on genotype data. Consequently, eight genetically independent markers were selected. The polymorphic information content values of the markers ranged from 0.269 to 0.764, with an average of 0.409. The markers could distinguish among 20 L. edodes cultivars and produced highly repeatable and reproducible results. The markers developed in this study will enable the precise identification of L. edodes cultivars, and may be useful for protecting breeders' rights.

• Horticultural Science & Technology
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• v.33 no.5
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• pp.722-729
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• 2015

Powdery mildew (PM) caused by Podosphaera aphanis is a major disease that can result in significant yield losses in strawberry (Fragaria ${\times}$ ananassa Duchesne). For preventing PM, pesticides are usually applied in strawberry. In this study, molecular markers were developed to increase breeding efficiency of PM-resistance cultivars by marker-assisted selection (MAS). An $F_2$ population derived from a cross between PM-resistance 'Seolhyang' and PM-susceptibility 'Akihime' was evaluated for disease resistance to PM and RAPD (random amplification of polymorphic DNA)-BSA (bulked segregant analysis). Among 200 RAPD primers tested, OPE10 primer amplified a 311bp-band present in with 331bp. Sequence alignment performed for searching polymorphisms and six single nucleotide polymorphism (SNP) were found in amplified regions. To develop polymorphic marker for distinguishing between resistant and susceptible, RAPD was converted to cleaved amplified polymorphic sequence (CAPS) marker. Among restriction enzymes associated with six SNPs, Eae I (Y/GGCCR) was successfully digested to 231bp in susceptible. The results suggest that the selected CAPS marker could be used for increasing efficiency of selecting powdery mildew resistant strawberry in breeding system.

• Journal of Forest and Environmental Science
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• v.25 no.1
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• pp.57-65
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• 2009

Ficus deltoidea Jack is an important and popular medicinal plant species found in the Malaysia. Plants are being collected and used based on morphology and authentication to prevent adulteration is not in practice. In this study, twenty-six accessions of F. deltoidea Jack were collected from Kelantan and Terengganu states of Peninsular Malaysia to examine their genetic similarities and differences using randomly amplified polymorphic DNA (RAPD) technique. Out of 20 arbitrary primers, two primers (D-10 and D-11) were selected which produced reliable DNA polymorphism. D-10 and D-11 primers generated 138 RAPD bands ranging from 250 bp to 3000 bp. Ninety-nine of them were polymorphic loci (72%) and thirty-nine were nonpolymorphic loci (28%). A total of 56 bands with polymorphic loci were amplified with primer D-10 and analyzed by cluster analysis and UPGMA to present a dendrogram depicting the degree of genetic relationship among 26 accessions. Eight RAPD markers were sequenced to determine their identity. RAPD analysis showed the genetic diversity among 26 accessions of F. deltoidea Jack. The RAPD profile and RAPD marker sequences reported in this paper could be used in plant and/or plant material authentication. This study also suggested that RAPD can be a useful technique to study DNA polymorphism in F. deltoidea Jack.

• Journal of Life Science
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• v.16 no.2 s.75
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• pp.219-224
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• 2006

Inter-simple sequence repeat (ISSR) analysis were used to assess genetic diversity among 18 genotypes of watermelon (Citrullus lanatus Thunb.) including breeding lines and commercial varieties. The 21 ISSR primers selected from 100 primers were showed the amplification of 105 reproducible fragments ranging from about 200 bp to 5000 bp. A total of 58 DNA fragments were polymorphic with an average 2.7 polymorphic bands per primer. The polymorphic primers were divided into 18 anchored primers and 3 non anchored primers. All of the anchored primers were di-nucleotide repeat motif, and was more polymorphic than non anchored primers. Eighteen watermelon genotypes were classified into two large groups. Clustering was in some accordance with the division of fruit shape into 18 watermelon. Therefore, ISSR markers may be suitable for variety discrimination and for constructing a linkage map of watermelon.

• Korean Journal of Weed Science
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• v.18 no.1
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• pp.76-83
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• 1998

Echinochloa species maintained by selling for more than 10 years were classified using random amplified polymorphic DNAs(RAPDs) analysis. Seventy-four decamer of randomly sequence markers were used to classify intraspecific variation irt Echinochloa species. The number of amplification products increased with increasing GC content of the primer in the range between 60% and 70% GC. Single-base substitutions of a primer altered amplification, providing new polymorphisms. The size of amplified DNA was mostly between 0.40kbp and 1.4kbp with the most common bands at 1.1kbp. Echinochloa species were detected with 6 primers which generated 26 polymorphic amplified DNAs. By hierarchical cluster analysis, Echinochloa species collected in Korea were divided into three groups. These results revealed that RAPD markers are useful tools for the determination of genetic variations in Echinochloa species.

• Journal of Plant Biotechnology
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• v.33 no.4
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• pp.289-295
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• 2006

This study was carried out to evaluate the suitability of simple sequence repeat (SSR) markers for varietal identification and genetic diversity in 28 commercial tomato varieties. The relationship between marker genotypes and 28 varieties was analyzed. Of the 219 pairs of SSR primers screened against ten tomato varieties, 18 pairs were highly polymorphic with polymorphism information content (PIC) ranging from 0.467 to 0.800. Among the polymorphic loci, two to nine SSR alleles were detected for each locus with an average of 3.3 alleles per locus. Genetic distances were estimated according to Jaccard's methods based on the probability that the amplified fragment from one genotype would be present in another genotype. These varieties were categorized into cherry and classic fruit groups corresponding to varietal types and genetic distance of cluster ranging from 0.35 to 0.97. The phonogram discriminated all varieties by marker genotypes. The SSR markers proved to be useful variety identification and genetic resource analysis of tomato.

• Journal of Forest and Environmental Science
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• v.35 no.2
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• pp.90-101
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• 2019

Biodiversity refers to the total number and variation among species of flora and fauna of an area. Due to tremendous biotic especially anthropogenic pressure these natural resources are being vanishing. In present study genetic diversity among accessions of Thamnocalamus spathiflorus was evaluated. A total of 51 vegetative characters and 42 primers (10-mer) were screened. Out of 42 screened primers, 28 polymorphic primers were selected for further analysis. A total of 263 bands were recorded as polymorphic whereas 48 bands were monomorphic. The resolving power (Rp) of 28 Randomly Amplified Polymorphic DNA (RAPD) primers ranged from 4.6 (OPE08) to 17.6 (OPA11). The polymorphic information content (PIC) value ranged from 0.21 (OPAH09) to 0.44 (OPG02). The result revealed high degree of genetic relatedness (56 to 80%). Cluster analysis revealed two major clusters both for morphology as well as RAPD. Unlike morphological characterization, the accession (D5) from Bahli, Rampur, Shimla (H.P.) was clustered separately from the others in RAPD cluster analysis. Accessions with closed locality grouped together through RAPD marker system however analogy was recorded for morphological traits. The study conducted reflects the utility of RAPD technique for species identification and phylogenetic studies in bamboo for conducting bamboo breeding program.

• Journal of Forest and Environmental Science
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• v.25 no.2
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• pp.93-100
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• 2009

In Malaysia, Labisia pumila Benth & Hook f, popularly known as 'Kacip Fatimah' has been used traditionally to treat various elements of the woman's health in Malay community. The objective of this study was to develop randomly amplified polymorphic DNA (RAPD) based DNA markers for the identification of L. pumila and to distinguish its three varieties from each other. Total DNA from nine accessions of L. pumila was extracted by CTAB method and polymerase chain reactions (PCR) were carried out to amplify the segments of DNA using different primers to develop DNA barcode using RAPD technique. To find out variety-specific DNA marker/s, twenty different 10-mer primer sequences with annealing temperature from 36-$40^{\circ}C$ were evaluated in triplicate. Out of 20 random primers, two primers (OPA-1 and OPA-2/A10) were selected which produced reliable RAPD band patterns. To have DNA based handle, two RAPD amplification products were cloned and sequenced to determine the identity of the DNA. RAPD analysis using two random primers generated 72 discrete bands ranging in size 200 bp-3,000 bp. Fifty nine of these were polymorphic loci (82%) and thirteen were non-polymorphic loci (18%). A total of 32 bands polymorphic loci (72%) were amplified with primer OPA-1 and analyzed by cluster analysis and UPGMA (Unweighted Pair Group Method with Arithmetic) to present a dendogram depicting the degree of genetic relationship among nine accessions of L. pumila. Our results shows the reasonable genetic diversity among the L. pumila varieties and within varieties; and two RAPD marker sequences obtained could be used to identify L. pumila at species level.