• Title/Summary/Keyword: Molecular identification of hybrids

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Molecular Identification and Effects of Temperature on Survival and Growth of Hybrids between Haliotis gigantea Gmelin (♀) and Haliotis discus hannai Reeve (♂)

  • An, Hye Suck;Han, Jong Won;Hwang, Hyun-Ju;Jeon, Hancheol;Jung, Seung-Hyun;Jo, Seonmi;Choi, Tae-Young;Hyun, Young Se;Song, Ha Yeun;Whang, Ilson
    • Journal of Marine Life Science
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    • v.2 no.2
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    • pp.83-89
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    • 2017
  • In abalones, interspecific hybridization has been suggested as a possible means to increase production and desired traits for the industry. In Korea, Haliotis gigantea is considered a species with a larger size and higher temperature tolerance than H. discus hannai. However, H. discus hannai is considered the most valuable and popular fishery resource due to its better acceptance and higher market prices. Thus, viable interspecific hybrids have been produced by artificial inseminating H. gigantea eggs with H. discus hannai sperm. However, the reciprocal hybrid cross was not successful. In this study, the hybridity and the growth and thermal tolerance performance of the interspecific hybrids were examined. A combination of various assays revealed maximum growth occurrence at 21℃ and the higher growth rate in the hybrids than that of H. discus hannai parent. In addition, the growth and survival at high-temperature (28℃) of the hybrids was equivalent to that of the highly tolerant H. gigantea parent, suggesting new possibilities to overcome the mass mortality in H. discus hannai during high temperature periods of summer season in Korea. Furthermore, the induced interspecific hybrid status was confirmed by the presence of species-specific bands for each parental species of the random amplified polymorphic DNA (RAPD) profiles using universal rice primer (URP), which could be used as speciesspecific markers to distinguish the hybrids and their parental species.

Production and Molecular Cytogenetic Identification of Wheat-Barley Hybrids and Translocations

  • Lang, Marta-Molnar;Gabriella Linc;Jozsef Sutka
    • Journal of Plant Biotechnology
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    • v.1 no.1
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    • pp.8-12
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    • 1999
  • New winter wheat winter barley hybrids were produced (Mv9 kr1 Igri, Mv9 kr1 Osnova, Asakazekomugi Manas). The wheat-barley hybrids showed entire male sterility and were multiplied in tissue culture. Chromosome configurations were studied with GISH in meiosis in the Mv9 krl x Igri hybrid and in its progenies multiplied in vitro. Chromosome pairing between wheat and barley has been observed in some cells in the hybrids multiplied in vitro. Backcross plants with 43 and 44 chromosomes were selected with the aim of developing new wheat-barley addition lines. Wheat-barley translocations were demonstrated with GISH in backcross progenies originating from in vitro regenerated wheat (Triticum aestivum L. cv. Chinese Spring) x barley (Hordeum vulgare L. cv. Betzes) hybrids. Five different translocations were observed. Sequential N-banding and GISH analyses were performed to further identify the translocations. The N-banding pattern of the Robertsonian translocation suggests that this chromosome consists of the short arm of barley chromosome 4H translocated to the long arm of 2B of wheat. Plants with four different homozygous translocations were selected from the following BC2F3 generation.

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Molecular characterization of reciprocal crosses of Aerides vandarum and Vanda stangeana (Orchidaceae) at the protocorm stage

  • Kishor, Rajkumar;Devi, H.S.;Jeyaram, K.;Singh, M.R.K.
    • Plant Biotechnology Reports
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    • v.2 no.2
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    • pp.145-152
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    • 2008
  • Aerides vandarum and Vanda stangeana are two rare and endangered vandaceous orchids with immense floricultural traits. The intergeneric hybrids were synthesized by performing reciprocal crosses between them. In vitro germination response of the immature hybrid embryos was found to be best on half-strength Murashige and Skoog medium supplemented with 20% (v/v) coconut water/liquid endosperm from tender coconut. Determination of hybridity was made as early as the immature seeds or embryos germinated in vitro, using randomly amplified polymorphic DNA (RAPD) markers. Out of 15 arbitrarily chosen decamer RAPD primers, two were found to be useful in amplification of polymorphic bands specific to the parental species and their presence in the reciprocal crosses. However, a decisive profile that can identify the reciprocal crosses could not be provided by RAPD. Amplification of the trnL-F non-coding regions of chloroplast DNA of the parent species and hybrids aided easy identification of the reciprocal crosses from the fact that maternal inheritance of chloroplast DNA held true for these intergeneric hybrids. Subsequent restriction digestion of the polymerase chain reaction (PCR) amplified trnL-F non-coding regions of chloroplast DNA also consolidated the finding. Such PCR-based molecular markers could be used for early determination of hybridity and easy identification of the reciprocal crosses.

Identification of Hybrid between the Tiger Grouper Epinephelus fuscoguttatus and the Giant Grouper E. lanceolatus by Analyzing COX I and RAG 2 Sequences (COX I 및 RAG 2 유전자 염기서열 분석에 의한 tiger grouper Epinephelus fuscoguttatus와 giant grouper E. lanceolatus 간 잡종의 동정)

  • Kim, Keun-Sik;Lee, Hyo-Ryeon;Sade, Ahemad;Bang, In-Chul
    • Korean Journal of Ichthyology
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    • v.26 no.1
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    • pp.70-73
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    • 2014
  • Interspecific hybrids between tiger grouper Epinephelus fuscoguttatus and giant grouper E. lanceolatus were genetically identified based on the partial sequence analysis of mitochondrial cytochrome c oxidase I (COX I) gene and nuclear recombination activating gene 2 (RAG 2) gene. Out of 585 base positions of RAG 2, a total of five nucleotide substitutions were detected between the two parental species (E. fuscoguttatus and E. lanceolatus). The hybrids had two distinct types of RAG 2 sequences corresponding to those of both parental species. Mitochondrial COX I gene sequencing showed that hybrids had sequences identical to E. fuscoguttatus. Molecular data clearly demonstrate that hybridization does occur between E. fuscoguttatus and E. lanceolatus, but with E. fuscoguttatus as the maternal parent.

Screening and classification of mulberry silkworm, Bombyx mori based on thermotolerance

  • Chandrakanth, Nalavadi;Moorthy, Shunmugam M.;Ponnuvel, Kangayam M.;Sivaprasad, Vankadara
    • International Journal of Industrial Entomology and Biomaterials
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    • v.31 no.2
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    • pp.115-126
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    • 2015
  • The tropical climate prevailing in India adversely affects temperate bivoltine silkworm rearing and causes crop loss especially during summer. Identification of high temperature tolerant bivoltine breeds by screening for thermotolerance in the silkworm, Bombyx mori (Lepidoptera: Bombycidae) is an essential prerequisite for the development of thermotolerant bivoltine breeds / hybrids. Therefore, in this study, 20 silkworm breeds were reared at different temperatures (25 ± 1℃,32 ± 1℃, 34 ± 1℃ and 36 ± 1℃) for 6 h every day from 3rd d of 5th instar to till spinning. Significant differences (p < 0.01) were found among all the rearing traits over temperature. Based on pupation percentage, SK4C and BHR3 were identified as thermotolerant bivoltine breeds. Hierarchical clustering analysis based on rearing traits at tested temperatures grouped 20 silkworm breeds in four clusters which included one cluster each of susceptible and tolerant, and two clusters of moderately tolerant silkworm breeds. This suggests that clustering based on rearing data at high temperatures by using Euclidean distance can be an effective approach in classifying the silkworm breeds on their thermotolerance capacity. The identified breeds would be used for development of thermo tolerant bivoltine silkworm breeds / hybrids.

Gene encoding prolactin of red-spotted grouper, Epinephelus akaara, and its application as a molecular marker for grouper species identification

  • Bok-Ki Choi;Gyeong-Eon Noh;Yeo-Reum Kim;Jun-Hwan Byun;HanKyu Lim;Jong-Myoung Kim
    • Fisheries and Aquatic Sciences
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    • v.27 no.6
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    • pp.346-355
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    • 2024
  • Groupers are economically important species in the fishery and aquaculture industries in Asian countries. Various species of grouper, including hybrids, have been brought to market even without precise species identification. In this study, we analyzed the structure and expression profile of the gene encoding prolactin (PRL) in the red-spotted grouper Epinephelus akaara based on genomic DNA and cDNA templates. The results showed that the PRL gene consists of five exons encoding an open reading frame of 212 amino acids, including a putative signal peptide of 24 amino acids and a mature structural protein of 188 amino acids. It showed amino acid identities of 99% with Epinephelus coioides, 83% with Amphiprion melanopus, 82% with Acanthopagrus schlegelii, 75% with Oreochromis niloticus, 70% with Coregonus autumnalis, and 67% with Oncorhynchus mykiss, indicating its closer similarity to E. coioides and other groupers but marked distinction from non-teleost PRLs. PRL mRNA expression was detected mostly in the brain, including the pituitary gland, with little expression in other tissues. While the 5-exon structure of the PRL gene of red-spotted grouper and the exon sizes were conserved, the sizes of the introns, particularly the first intron, were markedly different among the grouper species. To examine whether these differences can be used to distinguish groupers of similar phenotypes, exon-primed intron-crossing analysis was carried out for various commercially important grouper species. The results showed clear differences in size of the amplified fragment encompassing the first intron of the PRL gene, indicating that this method could be used to develop species-specific markers capable of discriminating between grouper species and their hybrids at the molecular level.

Molecular Biological Species Identification of Imported Groupers(Epinephelus moara ♀×E. lanceolatus ♂) (수입산 바리과(Family Serranidae) 잡종 어류(Epinephelus moara ♀×E. lanceolatus ♂)의 분자생물학적 판별)

  • Kim, Yong Hwi;Park, Jong Yeon;Kim, Jae Hoon;Bang, In-Chul
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.53 no.4
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    • pp.566-571
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    • 2020
  • To classify a presumed hybrid of imported grouper species acquired from the National Fishery Products Quality Management Service, maternal and paternal lines were identified based on partial sequencing of mitochondrial cytochrome c oxidase 1 (co1) and nuclear recombination activation gene 1 (rag1) genes. The matrilineal species was identified as Epinephleus moara by a partial (760 bp) co1 sequence. Ambiguous sequences with base pairs belonging to E. moara or E. lanceolatus were found in a total of 15 different base pairs in the partial 1,159 bp of the rag1 gene, and the patrilineal species was found to be E. lanceolatus. Therefore, all of the groupers examined in the study were identified to be hybrids of E. moara and E. lanceolatus. In addition, a fast and convenient method using random amplification of polymorphic DNA (RAPD) was established for hybrid discrimination. Hybrids between E. moara ♀ and E. lanceolatus ♂ were identified through specific bands of 387 bp and 433 bp in PRIMER 6.

Identification of Toxic Chemicals Using Polypyrrole-Cyclodextrin Hybrids (폴리피롤-사이클로덱스트린 혼성체를 이용한 유해화합물질의 검출)

  • Bae, Joonwon
    • Applied Chemistry for Engineering
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    • v.30 no.2
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    • pp.186-189
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    • 2019
  • Polypyrrole is a typical electrical conducting polymer, which has an excellent charge transport property. Cyclodextrins are a group of toxic-free and cyclic oligosaccharide molecules, capable of capturing low molecular weight chemicals. Considering these advantages, hybrid materials of polypyrrole and cyclodextrin can be used to detect hazardous compounds. Cyclodextrin molecules can accommodate toxic chemicals by the formation of host-guest complexes and generate electric signals, which are effectively delivered by polypyrrole backbone. In this study, the polypyrrole/cyclodextrin hybrid material was prepared using a facile wet method and included into a hydrogel. Subsequently, it was applied to a simple sensor system with a gold-patterned electrode for the detection of potentially hazardous material, methyl paraben. Compared with pristine polypyrrole, it was found that the polypyrrole/cyclodextrin hybrid showed an improved performance. This study can be an example of using environmentally benign conducting polymer/cyclodextrin hybrids as sensing media.

Phylogenetic position of Neofinetia and Sedirea (Orchidaceae) and their species identification using the chloroplast matK and the nuclear ITS sequences (엽록체 matK 와 핵 ITS 염기서열을 이용한 나도풍란속 및 풍란속의 계통과 종동정)

  • Kim, Young-Kee;Jo, Sang Jin;Kim, Ki-Joong
    • Korean Journal of Plant Taxonomy
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    • v.44 no.1
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    • pp.39-50
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    • 2014
  • Phylogenetic positions of Sedirea and Neofinetia were addressed using the chloroplast matK and the nuclear ITS sequences. We also evaluate the usefulness of the makers for the identification of species and localities. Sedirea and Neofinetia form an independent monophyletic genus, respectively, in both matK and nuclear ITS trees. The sister genus of the Neofinetia was Vanda in both trees. In addition, our trees support the separate recognition of the Neofinetia from Vanda rather than the inclusion of Neofinetia into Vanda. The sister group of the Sedirea was (Dimorphorchis(Pteroceras(Saccolabiun+Phalaeonopsis))) clade. The Dimorphorchis was one of the most probable sister genus to the Sedirea. The sister group relationship between Sedirea and Aerides was suggested by their similar morphology, but not supported in molecular trees. The identification of species and localities of Neofinetia was possible using our two molecular markers. However, several pseudo-gene sequences are discovered from the public data base. In addition, the horizontal gene transfer of chloroplast genomes is frequent events in orchid hybrids. Therefore, we need a careful evaluation for the data prior to systematic use. Generation of sequence data from multiple accessions of a species may helpful to reduce these types of error.

Genome-wide analysis of heterosis-related genes in non-heading Chinese cabbage

  • Yi, Hankuil;Lee, Jeongyeo;Song, Hayong;Dong, Xiangshu;Hur, Yoonkang
    • Journal of Plant Biotechnology
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    • v.44 no.3
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    • pp.208-219
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    • 2017
  • Heterosis or hybrid vigor describes a phenomenon that superior phenotypes compared to the two parents are observed in the heterozygous $F_1$-hybrid plants. Identification and characterization of heterosis-related genes (HRGs) will facilitate hybrid breeding in crops. To identify HRGs in Brassica rapa, we analyzed transcriptome profiling using a Br300K microarray in non-heading Chinese cabbage at three developmental stages. A large number of genes were differentially expressed in $F_1$ hybrids and non-additive expression was prominent. Genes that are expressed specifically for $F_1$ hybrid at all three stages were Brassica-specific uncharacterized genes and several defense-related genes. Expression of several photosynthesis- and stress-related genes were also $F_1$ hybrid-specific. Thirteen NBS-LRR class genes showed high and specific expression in $F_1$ hybrid Shulu: some of them were characterized as defense genes in Arabidopsis, but most have not been. Further characterization of these defense-related genes in Brassica species and its application will be helpful for understanding the role of defense responses in heterosis. In addition, results obtained in this study will be valuable to develop molecular markers for heterosis and disease resistance in B. rapa.