• Title/Summary/Keyword: molecular breeding

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Interspecific hybridization in seahorses: artificially produced hybrid offspring of Hippocampus kuda and Hippocampus reidi

  • Han, Sang-Yun;Rho, Sum;Noh, Gyeong Eon;Kim, Jin-Koo
    • Fisheries and Aquatic Sciences
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    • v.21 no.5
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    • pp.11.1-11.8
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    • 2018
  • Interspecific hybridization experiments were conducted between the common seahorse Hippocampus kuda (male) and the slender seahorse H. reidi (female) during artificial rearing to develop a new aquarium fish with unique polyandrous mating. Molecular analysis via mitochondrial DNA (mtDNA) cytochrome b and nuclear DNA (ncDNA) ribosomal protein S7 gene supported the hybridization between the two species, and the hybrid also showed morphological characteristics of both species. Juveniles of H. kuda have dense melanophores on the whole body or only on the trunk and tail, whereas juveniles of H. reidi have thin melanophores on the whole body or present in stripes only along their prominent trunk and tail rings. However, all the hybrid juveniles had dense melanophores only on the tail, with the striped trunk rings, thus showing an intermediate pattern, and these patterns were limited to the fairly early stage of development (1-10 days old). In contrast, the two eye spines in the hybrid were apparent after 9 days old, which were not inherited from H. kuda (one eye spine), but from H. reidi (two eye spines). According to LOESS (local regression) analysis, the growth rate increased between 20 and 25 days, and the hybrids grew faster than H. kuda when they entered the explosive second phase of growth between 25 and 45 days for all the seahorses. This study highlights the hybridization between H. kuda and H. reidi may contribute to the improved taxonomic information of young seahorses.

Single Nucleotide Polymorphisms on Peroxisome Proliferator-activated Receptor Genes Associated with Fatness Traits in Chicken

  • Meng, H.;Zhao, J.G.;Li, Z.H.;Li, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.9
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    • pp.1221-1225
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    • 2005
  • The peroxisome proliferator-activated receptors (PPARs) are members of a superfamily of nuclear hormone receptors. Lots of studies in rodents and humans have shown that PPARs were involved in lipid metabolism and adipocyte differentiation. The main objective of this work was to detect the single nucleotide polymorphisms (SNPs) in whole coding regions of peroxisome proliferator-activated receptor alpha (PPAR-$\alpha$) and gamma (PPAR-$\gamma$) genes with approach of single strand conformation polymorphism (SSCP) in the chicken population of Arber Acres broiler, Hyline layer and three Chinese native breeds (Shiqiza, Beijing You, Bai'r). Two SNPs of C1029T and C297T were found in chicken PPAR-$\alpha$ and PPAR-$\gamma$ genes respectively and each SNP found three genotypes in the experimental populations. The results showed that the distribution frequency of 3 genotypes in Arber Acres broiler, Hyline layer and Chinese native breeds had significant differences on the PPAR-$\alpha$ and PPAR-$\gamma$ gene respectively (p<0.01). Furthermore, in the PPAR-$\alpha$ gene, the results of least square estimation for genotypes and body composition traits showed the BB genotype birds had higher abdominal fat weight (AFW) and percentage of abdominal fat (AFP) than AA genotype birds (p<0.05). From these we conjecture the PPAR-$\alpha$ and PPAR-$\gamma$ genes were suffered intensive selection during the long term commercial breeding and the PPAR-$\alpha$ gene may be a major gene or linked to the major genes that impact chicken fat metabolism and the SNPs could be used in molecular assistant selection (MAS) as a genetic marker for the chicken fatness traits.

Barbigerone Inhibits Tumor Angiogenesis, Growth and Metastasis in Melanoma

  • Yang, Jian-Hong;Hu, Jia;Wan, Li;Chen, Li-Juan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.167-174
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    • 2014
  • Tumor angiogenesis, growth and metastasis are three closely related processes. We therefore investigated the effects of barbigerone on all three in the B16F10 tumor model established in both zebrafish and mouse models, and explored underlying molecular mechanisms. In vitro, barbigerone inhibited B16F10 cell proliferation, survival, migration and invasion and suppressed human umbilical vascular endothelial cell migration, invasion and tube formation in concentration-dependent manners. In the transgenic zebrafish model, treatment with $10{\mu}M$ barbigerone remarkably inhibited angiogenesis and tumor-associated angiogenesis by reducing blood vessel development more than 90%. In vivo, barbigerone significantly suppressed angiogenesis as measured by H and E staining of matrigel plugs and CD31 staining of B16F10 melanoma tumors in C57BL/6 mice. Furthermore, it exhibited highly potent activity at inhibiting tumor growth and metastasis to the lung of B16F10 melanoma cells injected into C57BL/6 mice. Western blotting revealed that barbigerone inhibited phosphorylation of AKT, FAK and MAPK family members, including ERK, JNK, and p38 MAPKs, in B16F10 cells mainly through the MEK3/6/p38 MAPK signaling pathway. These findings suggested for the first time that barbigerone could inhibit tumor-angiogenesis, tumor growth and lung metastasis via downregulation of the MEK3/6/p38 MAPK signaling pathway. The findings support further investigation of barbigerone as a potential anti-cancer drug.

Agrobacterium tumefaciens-mediated Transformation in Colletotrichum falcatum and C. acutatum

  • Maruthachalam, Karunakaran;Nair, Vijayan;Rho, Hee-Sool;Choi, Jae-Hyuk;Kim, Soon-Ok;Lee, Yong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.18 no.2
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    • pp.234-241
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    • 2008
  • Agrobacterum tumefaciens-mediated transformation (ATMT) is becoming an effective system as an insertional mutagenesis tool in filamentous fungi. We developed and optimized ATMT for two Colletotrichum species, C. falcatum and C. acutatum, which are the causal agents of sugarcane red rot and pepper anthracnose, respectively. A. tumefaciens strain SK1044, carrying a hygromycin phosphotransferase gene (hph) and a green fluorescent protein (GFP) gene, was used to transform the conidia of these two Colletotrichum species. Transformation efficiency was correlated with co-cultivation time and bacterial cell concentration and was higher in C. falcatum than in C. acutatum. Southern blot analysis indicated that about 65% of the transformants had a single copy of the T-DNA in both C. falcatum and C. acutatum and that T-DNA integrated randomly in both fungal genomes. T-DNA insertions were identified in transformants through thermal asymmetrical interlaced PCR (TAIL-PCR) followed by sequencing. Our results suggested that ATMT can be used as a molecular tool to identify and characterize pathogenicity-related genes in these two economically important Colletotrichum species.

Development of EST-SSR markers for genetic diversity analysis in little millet (Panicum sumatrense) genetic resources

  • Lee, Myung-Chul;Choi, Yu-Mi;Lee, Sukyeung;Yoon, Hyemyeong;Oh, Sejong
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.74-74
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    • 2018
  • Little millet (Panicum sumatrense) is well known for its salt and drought stress tolerance and high nutritional value, but very limited knowledge of genetic variation and genomic information is available. This study was to develop highly polymorphic EST-SSR markers based on cross-species transferability of derived SSRs from switchgrass EST databases and characterize newly developed EST - SSRs to better understand the genetic diversity of collected 37 germplasm accessions of little millet. A total of 779 primer pairs were designed from the 22,961 EST sequences of switchgrass (Pancium virgatum), of which 48 EST - SSR markers were developed based on the trials of transferability of these primers in little millet. The EST - SSR amplicons showed reproducible single band polymorphism and produced a total of 160 alleles with an average of 3.3 alleles per locus in 37 accessions of little millet. T he average values of expected and observed heterozygosities were 0.266 and 0.123, respectively. T he polymorphic information content (PIC) values were observed in range of 0.026 to 0.549 with an average of 0.240. The genetic relatedness among the little millet accessions was evaluated by neighbor-joining dendrogram, which grouped all accessions into two distinct groups. The validation thus demonstrated the utility of the switchgrass EST - SSR markers in assessing genomic relationships in little millet. T he findings from this study could be useful for designing strategies for the identification of diverse germplasm for conservation and future molecular breeding programs for little millet.

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Genetic Diversity and Population Structure of Spiraea prunifolia for. simpliciflora by Inter-Simple Sequence Repeats (조팝나무의 유전적 다양성과 집단구조 분석을 위한 ISSR 분석)

  • Huh, Man-Kyu
    • Journal of Life Science
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    • v.19 no.9
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    • pp.1183-1189
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    • 2009
  • 85 individual Spiraea prunifolia for. simpliciflora (Rosaceae) were sampled to examine the genetic diversity and population structure of S. prunifolia for. simpliciflora populations. Inter-simple sequence repeats (ISSR) produced 65 polymorphic loci and identified 78 ISSR genotypes. Three multilocus genotypes were shared by more than one plant within a population. Total genetic diversity values ($H_T$) and inter-locus variation in the within-population genetic diversity ($H_S$) were 0.293 and 0.183, respectively. On a per-locus basis, the proportion of total genetic variation due to differences among populations ($G_{ST}$) was 0.373. This indicated that about 37.3% of the total variation was among populations. ISSR markers are very effective in classifying natural population levels of S. prunifolia for. simpliciflora in Korea. In addition, insights into the relative gene diversity among and within populations of S. prunifolia for. simpliciflora would be useful in plant breeding and also for the development of strategies for ex situ conservation of plant genetic resources.

Morphological and Molecular Characterization of Thamnocalamus falconeri Hook f. ex. Munro

  • Tiwari, Chandrakant;Bakshi, Meena;Nautiyal, Subhash
    • Journal of Forest and Environmental Science
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    • v.31 no.3
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    • pp.214-224
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    • 2015
  • The economy of India and so also of many Asian countries depends on bamboos and their uses are not only in domestic items but also in rural housing and raw materials to several industries and germplasm characterization is an important link between the conservation and utilization of plant genetic resources. Classical taxonomic studies of the bamboos are based on floral morphology and growth habit, which can cause problems in identification due to erratic flowering coupled with different biotic agencies and environmental factors. Identification and genetic relationships among accessions of Thamnocalamus falconeri were investigated using morphology and random amplified polymorphic DNAs (RAPD) technique. Analysis started by using 51 vegetative characters and forty two 10-mer primers that allowed us to distinguish different genotypes hailing from different eco- zones of Garhwal Himalayas (India). The selected primers (12) were used for identification and for establishing a profiling system to estimate genetic diversity. A total of 79.33% polymorphism was estimated by using 12 selected primers. The genetic similar analysis was conducted based on binary digits i.e. presence (1) or absence (0) of bands, which revealed a wide range of variability among the species whereas genetic relatedness was quite high based on vegetative characters. Cluster analysis clearly showed two major clusters for both of the markers viz. morphology and RAPD belonging to 10 accessions of T. falconeri. Two major clusters were further divided into minor clusters. Cluster based on RAPD marker showed grouping of accessions of closed locality whereas analogy was reported for vegetative traits. The RAPD technique has the potential for use in species identification and genetic relationships studies of bamboo for breeding program.

Characterization of intraspecific hybrids by di-mon crossing in Pleurotus eryngii (Di-Mon 교잡법에 의한 큰느타리버섯 교잡주의 특성)

  • Shin, Pyung-Gyun;Yoo, Young-Bok;Kong, Won-Sik;You, Chang-Hyun;Oh, Se-Jong
    • Journal of Mushroom
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    • v.2 no.2
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    • pp.109-113
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    • 2004
  • Pleurotus eryngii is not edible and medicinal mushrooms indigenous to Korea. To improvement of strain suitable to the geographic setting of Korea, we are mated with 22 dikaryons and 47 monokaryons isolated from Pleurotus eryngii ASI 2547 by Di-Mon mating. 19 strains forming fruit body obtained from clamped 253 bred strains. 7 excellent strains are selected from 19 bred strains by various morphological features of fruit body. Among the selected 7 strains, H6 strain were identified into ASI 2547-like recombinant hybrids with URP uniprimer by RAPD analysis. This suggested that Di-Mon crossing is one of rapid and easy breeding method for strain improvement with molecular techniques.

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Identification of Different Species and Dultivars of Brassica by SDS-PAGE, Isozyme and Molecular Marker

  • Mukhlesur Rahman Md.;Hirata Yutaka
    • Journal of Plant Biotechnology
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    • v.7 no.1
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    • pp.27-35
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    • 2005
  • Eighty-five different cultivars of Brassica rapa, B. juncea, B. nap us, B. carinata, B. oleracea and hexaploid Brassica collected from Bangladesh, Japan, China and Denmark were analyzed by SDS-PAGE for seed and leaf protein variations, using esterase, acid phosphatase and peroxidase isozyme analysis. Ten polymorphic bands were identified from seed protein however no identifiable polymorphic band was found in the leaf protein. Polymorphic markers clearly distinguished the different Brassica species as well as yellow sarson (YS) and brown seeded (BS) cultivars of B. rapa. The $F_1$ cross between YS and brown seeded cultivars showed the existance of all poly-morphic bands of the respective parents. The Bangla-deshi and Japanese cultivars of B. rapa differed in the amount of seed protein. In the case of isozyme analysis, esterase showed the highest number of polymorphic bands (13) followed by acid phosphatase (9) and peroxidase (5). These polymorphic markers were very effec-tive for classification of all the species studied in this experiment. In parentage tests using isozymes, the hybridity of intra-and-interspecific crosses of almost all the seedlings could be identified from their respective cross combinations. Esterase polymorphism showed a clear differentiation between YS and BS types of B. rapa. In addition, two esterase polymorphic markers were iden ified to differentiate some cultivars of B. juncea. Segregation patterns in these two esterase bands showed a simple Mendelian monohybrid ratio of 3:1 in $F_2$, 1:1 in test cross and 1:0 in back cross progenies. No polymorphic band was identified to distinguish different cultivars of the same species by acid phosphatase or peroxidase. Polymerase Chain Reaction (PCR) was carried out with seed coat color specific marker of B. juncea. The yellow seeded cultivars produced a strong band at 0.5 kb and weak band 1.2 kb. In the addition of these two specific bands, Japanese yellow-seeded cultivars expressed two more weak bands at 1.0 kb and 1.1 kb. Where the brown seeded cultivars generated a single strong band at 1.1 kb. In segregating population, the yellow seed coat color marker segregated at a ratio 15 (brown) : 1 (yellow), indicating the digenic inheritance pattern of the trait.

High frequency plant proliferation via direct fronds regeneration of Korean endemic duckweed species (국내 자생 좀개구리밥 (Lemna paucicostata) 및 개구리밥(Spirodela polyrhiza)의 엽상체 증식을 통한 기내 식물체 대량 증식체계 확립)

  • Oh, Myung-Jin;Park, Jong-Mi;Ko, Suk-Min;Liu, Jang R.;Kim, Suk-Weon
    • Journal of Plant Biotechnology
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    • v.36 no.2
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    • pp.157-162
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    • 2009
  • High frequency plant proliferation system via direct frond regeneration of endemic duckweed plants Lemna paucicostata and Spirodela polyrhiza was established. Fronds of L. paucicostata and S. polyrhiza were able to multiply half-strength MS basal medium without plant growth regulators. However, addition of BA at a range of 1 to 3 mg/L was more effective than high concentration of BA treatments for fronds proliferation. Also half-strength MS salts was suitable for the fronds proliferation. Increase of salts concentration had inhibitory effect on fronds proliferation. Also the frequency of callus formation from fronds of L. paucicostata was 3.3%, when they cultured onto 1/2 MS medium supplemented with 1 mg/L of BA. Similarly the frequency of callus formation from S. polyrhiza was very low. After subculture of white globular structures derived from fronds of L. paucicostata, numerous globular somatic embryos and calluses were developed onto the surface of fronds. However these somatic embryos did not fully develop into normal plants when transferred to 1/2 MS basal medium. Therefore direct frond regeneration system was more efficient for mass proliferation of L. paucicostata and S. polyrhiza. The plant regeneration system of L. paucicostata and S. polyrhiza established in this study, might be applied to mass proliferation and genetic transformation for molecular breeding.