• Title/Summary/Keyword: Molecular Breeding

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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.

High-throughput identification of chrysanthemum gene function and expression: An overview and an effective proposition

  • Nguyen, Toan Khac;Lim, Jin Hee
    • Journal of Plant Biotechnology
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    • v.48 no.3
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    • pp.139-147
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    • 2021
  • Since whole-genome duplication (WGD) of diploid Chrysanthemum nankingense and de novo assembly whole-genome of C. seticuspe have been obtained, they have afforded to perceive the diversity evolution and gene discovery in the improved investigation of chrysanthemum breeding. The robust tools of high-throughput identification and analysis of gene function and expression produce their vast importance in chrysanthemum genomics. However, the gigantic genome size and heterozygosity are also mentioned as the major obstacles preventing the chrysanthemum breeding practices and functional genomics analysis. Nonetheless, some of technological contemporaries provide scientific efficient and promising solutions to diminish the drawbacks and investigate the high proficient methods for generous phenotyping data obtaining and system progress in future perspectives. This review provides valuable strategies for a broad overview about the high-throughput identification, and molecular analysis of gene function and expression in chrysanthemum. We also contribute the efficient proposition about specific protocols for considering chrysanthemum genes. In further perspective, the proper high-throughput identification will continue to advance rapidly and advertise the next generation in chrysanthemum breeding.

Tissues Expression, Polymorphisms of IFN Regulatory Factor 6 (IRF6) Gene and Their Associated with Immune Traits in Three Pig Populations

  • Liu, Yang;Xu, Jingeng;Fu, Weixuan;Weng, Ziqing;Niu, Xiaoyan;Liu, Jianfeng;Ding, Xiangdong;Zhang, Qin
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.2
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    • pp.163-169
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    • 2012
  • Interferon regulatory factor 6 (IRF6) gene is a member of the IRF-family, and plays functionally diverse roles in the regulation of the immune system. In this report, the 13,720 bp porcine IRF6 genomic DNA structure was firstly identified with a putative IRF6 protein of 467 amino acids. Alignment and phylogenetic analysis of the porcine IRF6 amino acid sequences with their homologies to other species showed high identity (over 96%). Tissues expression of IRF6 mRNA was observed by RT-PCR, the results revealed IRF6 expressed widely in eight tissues. One SNP (HQ026023:1383 G>C) in exon7 and two SNPs (HQ026023:130 G>A; 232 C>T) in the 5′ promoter region of porcine IRF6 gene were demonstrated by DNA sequencing analysis. A further analysis of SNP genotypes associated with immune traits including IFN-${\gamma}$ and IL10 concentrations in serum was carried out in three pig populations including Large White, Landraces and Songliao Black pig (a Chinese indigenous breed). The results showed that the SNP (HQ026023:1383 G>C) was significantly associated with the level of IFN-${\gamma}$ (d 20) in serum (p = 0.038) and the ratio of IFN-${\gamma}$ to IL10 (d 20) in serum (p = 0.041); The other two SNPs (HQ026023:130 G>A; 232 C>T) were highly significantly associated with IL10 level in serum both at the day 20 (p = 0.005; p = 0.001) and the day 35 (p = 0.004; p = 0.006). Identification of the porcine IRF6 gene will help our further understanding of the molecular basis of the IFN regulation pathway in the porcine immune response. All these results should indicate that the IRF6 gene can be regarded as a molecular marker associated with the IL10 level in serum and used for genetic selection in the pig breeding.

Development of a CAPS Marker Derived from the Pg-Actin Gene Sequences and RAPD Markers in Platycodon grandiflorum (도라지에서의 RAPD 마커 분석과 Actin 유전자 염기서열에서 유래한 CAPS 분자표지 개발)

  • Kim, Munhwi;Jeong, Eunah;Jeong, Jeongsu;Kwon, Soontae;Jeon, Ikjo;Jeong, Jeong Hag;Lee, Je Min;Yeam, Inhwa
    • Korean Journal of Plant Resources
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    • v.28 no.5
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    • pp.648-655
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    • 2015
  • Balloon flower (Platycodon grandiflorum A. DC.) is a perennial plant of mainly Campanulaceae family, which have been widely used as a food ingredient and herbal medicine in East Asia. Although demands on related products and yearly cultivation area for balloon flower are increasing, diverse fundamental technologies and molecular breeding studies are not very well supported in Platycodons. In this study, 30 random amplification of polymorphic DNA (RAPD) primers were test in an attempt to explore genetic diversities. In addition, sequences information of the actin gene, a well conserved gene encoding a globular protein that forms microfilaments, was retrieved and analyzed. Two actin homologs were recovered; 3.4 kb fragment is a Pg-actin and 1.4 kb fragment is a Pg-actin homolog with 28.6% similarity. We have confirmed that the Pg-actin gene is configured into 4 exons and 3 introns. A single nucleotide polymorphism (SNP), G↔A, was detected on the intron 3, which served as a target for the CAPS marker development. The marker Pg-Actin-Int3 was applied to 32 balloon flower accessions. Balloon flower DNA sequence information generated in this study is expected to contribute to the analysis and molecular breeding and genetic diversity analysis of balloon flowers.

Discrimination and Authentication of Eclipta prostrata and E. alba Based on the Complete Chloroplast Genomes

  • Kim, Inseo;Park, Jee Young;Lee, Yun Sun;Lee, Hyun Oh;Park, Hyun-Seung;Jayakodi, Murukarthick;Waminal, Nomar Espinosa;Kang, Jung Hwa;Lee, Taek Joo;Sung, Sang Hyun;Kim, Kyu Yeob;Yang, Tae-Jin
    • Plant Breeding and Biotechnology
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    • v.5 no.4
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    • pp.334-343
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    • 2017
  • Eclipta prostrata and E. alba are annual herbal medicinal plants and have been used as Chinese medicinal tonics. Both species are widely distributed in tropical and subtropical regions as well as in Korea. Both species have similar morphological features but E. alba has smoother leaf blade margins compared with E. prostrata. Although both species are utilized as oriental medicines, E. prostrata is more widely used than E. alba. Morphological semblances have confounded identification of either species. Here, we report the complete chloroplast genomes of both species to provide an authentication system between the two species and understand their diversity. Both chloroplast genomes were 151,733-151,757 bp long and composed of a large single copy (83,285-83,300 bp), a small single copy (18,283-18,346 bp), and a pair of inverted repeats (25,075-25,063 bp). Gene annotation revealed 80 protein coding genes, 30 tRNA genes and four rRNA genes. A phylogenetic analysis revealed that the genus Eclipta is grouped with Heliantheae tribe species in the Asteraceae family. A comparative analysis verified 29 InDels and 58 SNPs between chloroplast genomes of E. prostrata and E. alba. The low chloroplast genome sequence diversity indicates that both species are really close to each other and are not completely diverged yet. We developed six DNA markers that distinguish E. prostrata and E. alba based on the polymorphisms of chloroplast genomes between E. prostrata and E. alba. The chloroplast genome sequences and the molecular markers generated in this study will be useful for further research of Eclipta species and accurate classification of medicinal herbs.

Breeding of Flammulina velutipes Strains Adaptable to Elevated-temperature

  • Kong, Won-Sik;Cho, Yong-Hyun;Jhune, Chang-Sung;Yoo, Young-Bok;Kim, Kwang-Ho
    • Mycobiology
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    • v.32 no.1
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    • pp.11-16
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    • 2004
  • Winter mushroom, Flammulina velutipes, needs low temperature during its cultivation. To save on farm costs, especially during summer, a strain adaptable to a higher or elevated-temperature must be developed. At the start of breeding program, parental strains which could endure high temperature were obtained. Seuenty four dikaryotic strains were collected and divided into four groups according to the nature of temperature. They also had different fruiting temperature. Finally we selected three brown strains ASI 4048, 4057 and 4072, and collected their spores. These selected strains can germinate even at a high temperature of $32^{\circ}C$, which were dramatically higher than the other strains. Based on these results, the new white strain adapted to mid-temperature by backcross mating was developed. Molecular markers were applied to select white fruitbody producing strains without cultivation. They showed a specific band which co-segregated with brown fruitbody forming strains in $BC_1F_1$ progenies. Selected white strains were tested under several elevated temperature conditions.

Marker Development for Erect versus Pendant-Orientated Fruit in Capsicum annuum L.

  • Lee, Heung-Ryul;Cho, Myeong-Cheoul;Kim, Hyoun-Joung;Park, Sung-Woo;Kim, Byung-Dong
    • Molecules and Cells
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    • v.26 no.6
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    • pp.548-553
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    • 2008
  • The erect habit of fruit setting is a unique characteristic of ornamental peppers and wild pepper species. The erect habit is known to be controlled by the up locus on pepper (Capsicum annuum L.) chromosome 12. The result of a genetic analysis using Saengryeog 211 (pendant), Saengryeog 213 (erect), and their $F_1$ and $BC_1$ progeny demonstrated that up is a recessive gene. To develop an up-linked marker, bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) were employed using 108 $F_{2:3}$ individuals. The closest AFLP marker, $A2C7_{469}$, was located at a genetic distance of 1.7 cM from the up locus and was converted into a cleaved amplified polymorphic sequence (CAPS) marker. This marker was mapped at a genetic distance of 4.3 cM from the up locus. When the CAPS was applied to seven ornamental lines and 27 breeding lines with erect fruit, these genotypes of 28 lines were correctly predicted. Thus, the CAPS marker will be useful for marker-assisted selection (MAS) of pepper breeding lines with the up allele at the early seedling stage.

Molecular Screening of Blast Resistance Genes in Rice using SSR Markers

  • Singh, A.K.;Singh, P.K.;Arya, Madhuri;Singh, N.K.;Singh, U.S.
    • The Plant Pathology Journal
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    • v.31 no.1
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    • pp.12-24
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    • 2015
  • Rice Blast is the most devastating disease causing major yield losses in every year worldwide. It had been proved that using resistant rice varieties would be the most effective way to control this disease. Molecular screening and genetic diversities of major rice blast resistance genes were determined in 192 rice germplasm accessions using simple sequence repeat (SSR) markers. The genetic frequencies of the 10 major rice blast resistance genes varied from 19.79% to 54.69%. Seven accessions IC337593, IC346002, IC346004, IC346813, IC356117, IC356422 and IC383441 had maximum eight blast resistance gene, while FR13B, Hourakani, Kala Rata 1-24, Lemont, Brown Gora, IR87756-20-2-2-3, IC282418, IC356419, PKSLGR-1 and PKSLGR-39 had seven blast resistance genes. Twenty accessions possessed six genes, 36 accessions had five genes, 41 accessions had four genes, 38 accessions had three genes, 26 accessions had two genes, 13 accessions had single R gene and only one accession IC438644 does not possess any one blast resistant gene. Out of 192 accessions only 17 accessions harboured 7 to 8 blast resistance genes.

Pedigree Analysis of 17 High Quality Korean Rice Cultivars Using Web Database Systems

  • Yi, Gi-Hwan;Park, Dong-Soo;Chung, Eun-Sook;Song, Song-Yi;Jeon, Nam-Soo;Nam, Min-Hee;Kim, Doh-Hoon;Han, Chang-Deok;Eun, Moo-Young;Ku, Yeon-Chung
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.6
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    • pp.554-564
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    • 2006
  • It is important to understand pedigree of rice cultivars commonly used for breeding. In this paper, pedigree tables for tracking the pedigree of 17 representative rice cultivars recommended by Rural development Adminstration (RDA) were completed and analyzed using two kinds of web database system; 'IRIS' and 'RRDB'. Seven cultivars, namely, 'Sangmibyeo', 'Ilpumbyeo', 'Saegewhabyeo', 'Surabyeo', 'Shindongjinbyeo', 'Ilmibyeo' and 'Jungwhabyeo' had 'Koshihikari' on the pedigree of their ancestor. Besides 'Koshihikari', the most feguently used ancestral germplasms among the high quality rice cultivars were 'Fujisaka 5', 'Kameno o' and 'Asahi', 'Fujisaka 5' was used as ancestral parent in 12 out of 17 cultivars. Interestingly, 'Kameno o' was used in pedigree of 16 out of 17 high quality varieties and 'Asahi' was used in the ancestral pedigree of all 17 varieties. 'Hwayeongbyeo' was used as one of parent in the breeding of 'Dongjin 1', 'Hwabongbyeo', 'Saegewhabyeo' and 'Junambyeo'. 'Ilpumbyeo' was used in the breeding pathway of 'Junambyeo' and 'Saegewhabyeo', 'Mangeumbyeo' itself was not enlisted as one of high quality rice cultivars, but was used as a breeding parent of three high quality varieties, namely, 'Saegewhabyeo', 'Hwabongbyeo' and 'Nampyeongbyeo'. Incorporated with evaluation data, pedigree will provide a valuable chance to genealogical tracking of agronomic traits such as disease resistance, grain quality and etc.

Status of research on the sweetpotato biotechnology and prospects of the molecular breeding on marginal lands (고구마 생명공학연구 현황과 조건 불리지역 분자육종 전망)

  • Kim, Ho Soo;Yoon, Ung-Han;Lee, Chan-Ju;Kim, So-Eun;Ji, Chang Yoon;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.45 no.3
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    • pp.196-206
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    • 2018
  • Dramatic increase in global population accompanied by rapid industrialization in developing countries has led to serious environmental, food, energy, and health problems. The Food and Agriculture Organization of the United Nations has estimated world population will increase to 9.7 billion by 2050 and require approximately 1.7 times more food, and more than 3.5 times energy than that of today. Particularly, sweetpotato is easy to cultivate in unfavorable conditions such as heat, drought, high salt, and marginal lands. In this respect, sweetpotato is an industrially valuable starch crop. To replace crops associated with these food and energy problems, it is necessary to develop new crops with improved nutrients and productivity, that can be grown on marginal lands, including desertification areas using plant biotechnology. For this purpose, exploring useful genes and developing genetically modified crops are essential strategies. Currently, sweetpotato [Ipomoea batatas (L.) Lam.] have been re-evaluated as the best health food and industrial crop that produces starch and low molecular weight antioxidants, such as vitamin A, vitamin E, anthocyanins and carotenoids. This review will focus on the current status of research on sweetpotato biotechnology on omics including genome sequencing, transcriptome, proteomics and molecular breeding. In addition, prospects on molecular breeding of sweetpotato on marginal lands for sustainable development were described.