• Title/Summary/Keyword: Simple Sequence Repeat

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Improved characterization of Clematis based on new chloroplast microsatellite markers and nuclear ITS sequences

  • Liu, Zhigao;Korpelainen, Helena
    • Horticulture, Environment, and Biotechnology : HEB
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    • v.59 no.6
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    • pp.889-897
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    • 2018
  • Currently, there is a lack of genetic markers capable of effectively detecting polymorphisms in Clematis. Therefore, we developed new markers to investigate inter- and intraspecific diversity in Clematis. Based on the complete chloroplast genome of Clematis terniflora, simple sequence repeats were explored and primer pairs were designed for all ten adequate repeat regions (cpSSRs), which were tested in 43 individuals of 11 Clematis species. In addition, the nuclear ITS region was sequenced in 11 Clematis species. Seven cpSSR loci were found to be polymorphic in the genus and serve as markers that can distinguish different species and be used in different genetic analyses, including cultivar identification to assist the breeding of new ornamental cultivars.

Genetic Stability Studies in Micropropagated Date Palm (Phoenix dactylifera L.) Plants using Microsatellite Marker

  • Kumar, Nitish;Singh, Amritpal S.;Modi, Arpan R.;Patel, Armi R.;Gajera, Bhavesh B.;Subhash, Narayanan
    • Journal of Forest and Environmental Science
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    • v.26 no.1
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    • pp.31-36
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    • 2010
  • Sixteen microsatellite markers (simple sequence repeat (SSR) markers) were employed to examine the genetic stability of 27 randomly chosen date palm (Phoenix dactylifera L.) plants produced through somatic embryogenesis with upto forty two in vitro subcultures. No microsatellite DNA variation was observed among all micropropagated plants. Our results indicate that the micropropagation protocol used for rapid in vitro multiplication is appropriate and suitable for clonal propagation of date palm and corroborated that somatic embryogenesis can also be used as one of the safe modes for production of true-to-type plants of date palm. This is the first report on the use of microsatellite DNA markers to establish the genetic stability in micropropagated date palm plants.

Genomic Analyses of Toll-like Receptor 4 and 7 Exons of Bos indicus from Temperate Sub-himalayan Region of India

  • Malik, Y.P.S.;Chakravarti, S.;Sharma, K.;Vaid, N.;Rajak, K.K.;Balamurugan, V.;Biswas, S.K.;Mondal, B.;Kataria, R.S.;Singh, R.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.7
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    • pp.1019-1025
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    • 2011
  • Toll-like receptors (TLRs) play an important role in the recognition of invading pathogens and the modulation of innate immune responses in mammals. The TLR4 and TLR7 are well known to recognize the bacterial lipopolysaccharide (LPS) and single stranded (ssRNA) ligands, respectively and play important role in host defense against Gram-negative bacteria and ssRNA viruses. In the present study, coding exon fragments of these two TLRs were identified, cloned, sequenced and analyzed in terms of insertion-deletion polymorphism, within bovine TLRs 4 and 7, thereby facilitating future TLR signaling and association studies relevant to bovine innate immunity. Comparative sequence analysis of TLR 4 exons revealed that this gene is more variable, particularly the coding frame (E3P1), while other parts showed percent identity of 95.7% to 100% at nucleotide and amino acid level, respectivley with other Bos indicus and Bos taurus breeds from different parts of the world. In comparison to TLR4, sequence analysis of TLR7 showed more conservation among different B. indicus and B. taurus breeds, except single point mutation at 324 nucleotide position (AAA to AAM) altering a single amino acid at 108 position (K to X). Percent identity of TLR7 sequences (all 3 exons) was between 99.2% to 100% at nucleotide and amino acid level, when compared with available sequence database of B. indicus and B. taurus. Simple Modular Architecture Research Tool (SMART) analysis showed variations in the exon fragments located in the Leucine Rich Repeat (LRR) region, which is responsible for binding with the microbial associated molecular patterns and further, downstream signaling to initiate anti-microbial response. Considering importance of TLR polymorphism in terms of innate immunity, further research is warranted.

Molecular Authentication and Genetic Polymorphism of Korean Ginseng (Panax ginseng C. A. Meyer) by Inter-Simple Sequence Repeats (ISSRs) Markers (ISSRs 마크에 의한 고려 인삼의 분자적 인증과 유전적 다형현상)

  • Bang, Kyong-Hwan;Lee, Sung-Woo;Hyun, Dong-Yun;Cho, Joon-Hyeong;Cha, Seon-Woo;Seong, Nak-Sul;Huh, Man-Kyu
    • Journal of Life Science
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    • v.14 no.3
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    • pp.425-428
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    • 2004
  • Molecular authentication and genetic polymorphism of Korean ginseng cultivars and accessions were investigated using ISSR (inter-simple sequence repeat amplification) markers. Five primers among 56 produced clear and reproducible DNA fragments among seven cultivars and accessions. A total of 43 bands ranging from 250 bp to 1,700 bp from five primers were scored. Average number of bands per primer was 8.6 and only nine bands were polymorphic across the six Panax ginseng from Korea. Especially Chunpoong cultivar exhibited the highest level of polymorphism, whereas other accessions did not showed almost any polymorphism. Consequently, these ISSR markers will be available to differentiate Chunpoong cultivar from other major Korean ginseng cultivars and accessions, such as Yunpoong, Hwangsukjong and Jakyungjong, at the DNA level.

Morphological Characteristics and Genetic Diversity Analysis of Platycodon grandiflorum (Jacq.) A. DC Determined Using SSR Markers (도라지 수집종의 형태적 특성과 SSR마커에 의한 유연관계 분석)

  • Um, Yurry;Lee, Yi;Jin, Mei-Lan;Lee, Dae Young;Lee, Jae Won;Kim, Geum Soog;Kim, Chang Kug;Hong, Chang Pyo;Kim, Ok Tae
    • Korean Journal of Medicinal Crop Science
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    • v.24 no.1
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    • pp.55-61
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    • 2016
  • Background : Plant breeding requires the collection of genetically diverse genetic resources. Studies on the characteristics of Platycodon grandiflorum resources have not been carried out so far. The present study was carried out to discriminate P. grandiflorum based on morphological characteristics and genetic diversity using simple sequence repeat (SSR) markers. Methods and Results :We collected 11 P. grandiflorum cultivars: Maries II, Hakone double white, Hakone double blue, Fuji white, Fuji pink, Fuji blue, Astra white, Astra pink, Astra blue, Astra semi-double blue and Jangbaek. Analyses of the morphological characteristics of the collection were conducted for aerial parts (flower, stem and leaf) and underground parts (root). Next, the genetic diversity of all P. grandiflorum resources was analyzed using SSR markers employing the DNA fragment analysis method. We determined that the 11 P. grandiflorum cultivars analyzed could be classified by plant length, leaf number and root characteristic. Based on the genetic diversity analysis, these cultivars were classified into four distinct groups. Conclusions : These findings could be used for further research on cultivar development using molecular breeding techniques and for conservation of the genetic diversity of P. grandiflorum. Moreover, the markers could be used for genetic mapping of the plant and marker-assisted selection for crop breeding.

Transcriptome analysis of a medicinal plant, Pistacia chinensis

  • Choi, Ki-Young;Park, Duck Hwan;Seong, Eun-Soo;Lee, Sang Woo;Hang, Jin;Yi, Li Wan;Kim, Jong-Hwa;Na, Jong-Kuk
    • Journal of Plant Biotechnology
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    • v.46 no.4
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    • pp.274-281
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    • 2019
  • Pistacia chinensis Bunge has not only been used as a medicinal plant to treat various illnesses but its young shoots and leaves have also been used as vegetables. In addition, P. chinensis is used as a rootstock for Pistacia vera (pistachio). Here, the transcriptome of P. chinensis was sequenced to enrich genetic resources and identify secondary metabolite biosynthetic pathways using Illumina RNA-seq methods. De novo assembly resulted in 18,524 unigenes with an average length of 873 bp from 19 million RNA-seq reads. A Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation tool assigned KO (KEGG orthology) numbers to 6,553 (36.2%) unigenes, among which 4,061 unigenes were mapped into 391 different metabolic pathways. For terpenoid backbone and carotenoid biosynthesis pathways, 44 and 22 unigenes encode enzymes corresponding to 30 and 16 entries, respectively. Twenty-two unigenes encode proteins for 16 entries of the carotenoid biosynthesis pathway. As for the phenylpropanoid and flavonoid biosynthesis pathways, 63 and 24 unigenes were homologous to 17 and 14 entry proteins, respectively. Mining of simple sequence repeat identified 2,599 simple sequence repeats from P. chinensis unigenes. The results of the present study provide a valuable resource for in-depth studies on comparative and functional genomics to unravel the underlying mechanisms of the medicinal properties of Pistacia L.

Genetic diversity and population structure of Chinese ginseng accessions using SSR markers

  • An, Hyejin;Park, Jong-Hyun;Hong, Chi Eun;Raveendar, Sebastin;Lee, Yi;Jo, Ick-Hyun;Chung, Jong-Wook
    • Journal of Plant Biotechnology
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    • v.44 no.3
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    • pp.312-319
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    • 2017
  • The need to preserve and use plant genetic resources is widely recognized, and the prospect of dwindling plant genetic diversity, coupled with increased demands on these resources, has made them a topic of global discussion. In the present study, the genetic diversity and population structure of 73 ginseng accessions collected from six regions in China were analyzed using eight simple sequence repeat (SSR) markers. Major allele frequencies ranged between 0.38 ~ 0.78, with a mean allele frequency value of 0.571. The number of alleles discovered ranged from 3 to 10 per accession, with a mean number of 7; 56 alleles were discovered in total. Gene diversity (GD) and polymorphic information content (PIC) values were similar to each other, and they ranged from 0.36 ~ 0.77 (mean 0.588) and 0.33 ~ 0.74 (mean 0.548), respectively. Accessions were divided into three clusters based on their phylogenetic relationships and genetic similarities, and although the populations were similar, they were not classified according to the region. Regional genetic diversity was also similar, with slight differences observed based on the number of accessions per region. It is expected that the findings of the present study can provide basic data for future studies on ginseng genetic diversity and for breeding ginseng cultivars.

Chromosome-specific polymorphic SSR markers in tropical eucalypt species using low coverage whole genome sequences: systematic characterization and validation

  • Patturaj, Maheswari;Munusamy, Aiswarya;Kannan, Nithishkumar;Kandasamy, Ulaganathan;Ramasamy, Yasodha
    • Genomics & Informatics
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    • v.19 no.3
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    • pp.33.1-33.10
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    • 2021
  • Eucalyptus is one of the major plantation species with wide variety of industrial uses. Polymorphic and informative simple sequence repeats (SSRs) have broad range of applications in genetic analysis. In this study, two individuals of Eucalyptus tereticornis (ET217 and ET86), one individual each from E. camaldulensis (EC17) and E. grandis (EG9) were subjected to whole genome resequencing. Low coverage (10×) genome sequencing was used to find polymorphic SSRs between the individuals. Average number of SSR loci identified was 95,513 and the density of SSRs per Mb was from 157.39 in EG9 to 155.08 in EC17. Among all the SSRs detected, the most abundant repeat motifs were di-nucleotide (59.6%-62.5%), followed by tri- (23.7%-27.2%), tetra- (5.2%-5.6%), penta- (5.0%-5.3%), and hexa-nucleotide (2.7%-2.9%). The predominant SSR motif units were AG/CT and AAG/TTC. Computational genome analysis predicted the SSR length variations between the individuals and identified the gene functions of SSR containing sequences. Selected subset of polymorphic markers was validated in a full-sib family of eucalypts. Additionally, genome-wide characterization of single nucleotide polymorphisms, InDels and transcriptional regulators were carried out. These variations will find their utility in genome-wide association studies as well as understanding of molecular mechanisms involved in key economic traits. The genomic resources generated in this study would provide an impetus to integrate genomics in marker-trait associations and breeding of tropical eucalypts.

Evaluation of Genetic Diversity among Persimmon Cultivars (Diospyros kaki Thunb.) Using Microsatellite Markers (초위성 마커를 이용한 감(Diospyros kaki Thunb.)의 유연관계 분석)

  • Hwang, Ji-Hyeon;Park, Yu-Ok;Kim, Sung-Churl;Lee, Yong-Jae;Kang, Jum-Soon;Choi, Young-Whan;Son, Beung-Gu;Park, Young-Hoon
    • Journal of Life Science
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    • v.20 no.4
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    • pp.632-638
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    • 2010
  • The genetic diversity among 48 persimmon (Diospyros kaki Thunb.) accessions, indigenous in Korea and introduced from Japan and China, was evaluated by using simple sequence repeat (SSR) markers. From 20 SSR primer sets, a total of 114 polymorphic markers were detected among 12 pollination-constant non-astringent (PCNA), 13 pollination-variant non-astringent (PVNA), 15 pollination-variant astringent (PVA), and 8 pollination-constant astringent (PCA) cultivars. Analysis of pair-wise genetic similarity coefficient (Nei-Li) and unweighted pair-group method with arithmetic averaging (UPGMA) clustering revealed two main clusters and four subclusters for cluster I. The subclustering pattern was in accordance with the classification of persimmon cultivars based on the nature of astringency loss. Phenetic relationships among the subclusters showed a closer relatedness of the PCNA group with the PVNA group, and the PVA with the PCA group. Genetic similarity co-efficiency was 0.499 on average and the highest (0.954) similarity was observed between 'Cheongdo-Bansi' and 'Haman-Bansi'. The similarity was lowest (0.192) between 'Damopan'and 'Atago'. Identification of each cultivar with the execption of 'Cheongdo-Bansi' and 'Gyeongsan-Bansi' was possible based on the SSR fingerprints, suggesting that these SSR markers are a useful tool for protecting intellectual property on newly developed cultivars.