• Title/Summary/Keyword: Doubled haploid(DH)

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Development of Doubled-haploid Population and Construction of Genetic Map Using SSR Markers in Rice (벼의 Doubled-haploid 집단육성과 SSR 마커를 이용한 유전자 지도작성)

  • Kim, Kyung-Min;Nam, Wu-Il;Kwon, Yong-Sham;Sohn, Jae-Keun
    • Journal of Plant Biotechnology
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    • v.31 no.3
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    • pp.179-184
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    • 2004
  • A doubled-haploid (DH) population was developed through anther culture of F$_1$ plants obtained from a cross between a japonica cultivar, 'Nagdongbyeo', as male parent and a indica cultivar, 'Samgangbyeo', as female parent. Segregation modes for plant length, culm length, panicle length, third internode length, and days to heading in the DH lines showed nearly normal distribution with wide range of variation. A molecular map with 136 simple sequence repeat (SSR) markers was constructed using the DH population. The total map distance was 1,909 cM and the average interval of marker distance was 14 cM.

Use of Androgenesis in Haploid Breeding

  • Yi, Gihwan;Kim, Kyung-Min;Sohn, Jae-Keun
    • Current Research on Agriculture and Life Sciences
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    • v.31 no.2
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    • pp.75-82
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    • 2013
  • Haploids are plants with a gametophytic number of chromosomes in their sporophytes. Androgenesis occurs from asymmetric division of pollen grains into generative cells and vegetative cells, followed by re-entry of the vegetative cell during S-phase, which causes microspores progress into G2/M transition in culture. One of the most interesting features of haploids is the possibility to produce doubled haploid (DH) individuals. Doubled haploidy is extremely useful to plant breeders because it enables shortened breeding periods and efficiency in selection of useful recessive agronomic traits. Doubled-haploid technology is not only applicable to breeding, but also to transformation programs of desired genes. In addition to practical breeding programs, DH lines provide useful materials of fundamental genetics including exploitation of QTLs and genes conferred with various agronomic traits by establishing DH populations. This paper provides historical overviews on androgenesis and describes several mechanisms associated with pollen embryogenesis, including mode of actions in pollen embryogenesis, mechanisms of chromosome doubling and factors affecting androgenesis. We also discuss recent progress in application of haploids to breeding, genes associated with in vitro response and drawbacks to anther culture for application of doubled haploids in crop breeding.

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Genetic Ana1ysis for Rice Grain Properties Using a Doubled Haploid Population

  • Qin, Yang;Kim, Suk-Man;Sohn, Jae-Keun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.52 no.2
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    • pp.123-128
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    • 2007
  • Demand for high quality rice has always been a major factor in the international rice marketing. In the present study, doubled haploid (DH) population derived from anther culture of a Tongil/japonica hybrid was used for genetic analysis of rice grain quality. The average values of DH lines for grain weight, grain length and the ratio of grain length to width were near the mid-parent value. More than 40% DH lines showed transgressive segregation for grain weight, length, amylose and lipid content, but less than 10% DH lines observed on ratio of length to width and grain thickness were transgressive segregation. Correlation analysis between appearance qualities and physicochemical characters indicated that grain width and grain thickness both significantly and negatively correlated to protein and lipid content. A highly significant negative correlation between protein content and amylose content was observed.

Development of Clubroot Resistant Doubled-Haploid Inbred Lines in Kimchi Cabbage (Chinese Cabbage) (Brassica rapa L.)

  • Park, Suhyoung;Jang, Hayoung;Park, Min Young
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.37-37
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    • 2015
  • Kimchi cabbage (Chinese cabbage), radish and Cabbage are major Brassicaceae vegetables in Korea. Especially, we can easily develop whole plant from one microspore in Kimchi cabbage. To develop clubroot resistant doubled-haploid (DH) inbred lines, we pollinated a clubroot resistant turnip of 'IT 033820' with a Kimchi cabbage (Chinese cabbage) inbred of 'BP 079'. More than 85 DH inbred lines were developed from this combination. We screened about 400 materials including these DH inbred lines, commercial cultivars and breeding materials during 3 years using hydroponic system after inoculating single spore isolation race 4(SSI-04) inoculate. One inbred line derived from this combination selected as clubroot resistant and registered as 'Wonkyo20036ho'. We inoculated 26 DH inbred lines derived from 'Zoong-baek 2ho' using SSI-4, the percent of resistant plants varied from 0 to 83%. However the horticultural traits of highly resistant DH inbred line was poor. Thus we selected one DH line showing 77% resistant with yellow inner leaf and maid good head, was registered as 'Wonkyo20034ho'. Another DH inbred line derived from Korean variety of 'Wol-dong' showing 86% resistant was registered as 'Wonkyo20037ho'. Other DH inbred lines were derived from Chinese cultivar of 'Choon-hi-go-hang-wang' and 'Hwang-shim-zo48' showed 80 and 71% resistant, respectively, was also selected for registration. Even though DH inbred lines derived from turnip showed highly resistant to SSI-04 and provincial inoculate, they showed poor characteristics in horticultural traits. However, commercial seed companies showed interesting for adapting these DH inbred lines in commercial breeding.

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Quantitative trait loci (QTLs) detection for plant regeneration ability from seed culture in rice (Oryza sativa L.)

  • Liu, Meihan;Sohn, Jae-Keun
    • Journal of Plant Biotechnology
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    • v.39 no.3
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    • pp.169-174
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    • 2012
  • Quantitative trait loci (QTLs), which were related to the ability of callus induction and plant regeneration in seed culture of rice, were analyzed using a mapping population from a cross between the rice cultivars 'Samgang' (tongil type) and 'Nagdong' (japonica). A tongil type rice cultivar, 'Samgang' showed lower frequency (20%) of plant regeneration than that (35%) of japonica rice, 'Nagdong'. Transgressive segregations were observed for the ability of callus induction and plant regeneration from the seed-derived calli of 58 doubled haploid (DH) lines. The ability of plant regeneration of 58 doubled haploid lines showed a continuous distribution with comparatively wide range (10.0 to 66.7%) of variation. Composite interval mapping analysis was used to identify the QTLs controlling callus induction and plant regeneration ability. Four significant QTLs, qCWS6, qCWS8, qCWS9 and qCWS11, associated with callus weight per seed were detected on chromosomes 6, 8, 9, and 11 with LOD values of 3.30, 2.60, 2.70 and 2.43, explaining 36% of the total phenotypic variation. Three significant QTLs, qPR1, qPR6, and qPR11, for the ability of plant regeneration were located on chromosome 1, 6, and 11 at LOD score of 2.25, 2.15 and 2.55, accounting for 24 % of the total phenotypic variation. The present study should be useful for improving the efficiency of plant regeneration in tissue culture of indica rice by means of marker-assisted selection.

QTL Analysis of Protein Content in Double-haploid Lines of Rice

  • Qin, Yang;Kim, Suk-Man;Sohn, Jae-Keun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.54 no.2
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    • pp.165-171
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    • 2009
  • The objective of this study was to characterize the main-effect QTLs, epistatic QTLs and QTL-by-environment interactions (QE), which are involved in the control of protein content. A population of 120 doubled haploid (DH) lines derived from a cross between 'Samgang' and 'Nagdong', was planted and determined for protein content over three years. Based on the population and a genetic linkage map of 172 markers, QTL analysis was conducted by WinQTLcart 2.5 and QTLMAPPER. Three main-effect QTLs affecting protein content of brown rice were detected from 2004 to 2006 on chromosomes 1 and 11. The qPC11.2 was repeatedly detected across two years. Seven pairs of epistatic loci were identified on eight chromosomes for protein content and collectively explained 39.15% of phenotype variation. These results suggest that epistatic effects might be an even more important component of the genetic basis for protein content and that the segregation of the DH lines for protein content could be largely explained by a few main-effect QTLs and many epistatic loci.

Mapping of RFLP Markers Linked to Bacterial Blight Resistant Genes (Xa-1, Xa-3) in Rice (벼 흰잎마름병 저항성 유전자(Xa-1, Xa-3)연관 RFLP 마커 탐색)

  • 강현중;김현순;남정권;이영태;이승엽;김석동
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.48 no.6
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    • pp.419-423
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    • 2003
  • Bacterial blight caused by Xantomonas oryzae pv. oryzae is one of the most serious diseases of rice especially in southern area of Korea. Three races, $\textrm{K}_1$, $\textrm{K}_2$ and $\textrm{K}_3$, are the most dominant species. lo improve rice breeding efficiency using marker assisted selection, some RFLP markers were surveyed for polymorphism between resistant and susceptible to $\textrm{K}_1$ and $\textrm{K}_3$. And, 127 doubled-haploid (DH) lines derived from Milyang121/HRl1650-1-4-2 and 131 DH lines derived from Milyang123/HR10624-AC5 were evaluated to bacterial blight ($\textrm{K}_1$ and $\textrm{K}_3$). Milyang121 and HR10624-AC5 have Xa-1, resistant to $\textrm{K}_1$ race, and Milyang123 has Xa-3, resistant to $\textrm{K}_1$ and $\textrm{K}_3$ race. Three markers, RZ590, RZ536 and RG303, showing polymorphism between parents and resistance gene, Xa-1 and Xa-3, were analysed in the two combinations of DH lines. The segregation pattern of resistant DH population of Milyang123/HR10624-AC5 to susceptible showed 3:1 and 1:1 in $\textrm{K}_1$ and $\textrm{K}_3$ race. In three RFLP markers, RZ590 was linked to Xa-1 on chromosome 4, and RZ536 and RG303 were linked to Xa-3 on chromosome 11. The map distance between Xa-1 and RZ590 was 3.1cM on chromosome 4, and Xa-3 and RZ536/RG303 were 7.6/16.0cM on chromosome 11, respectively. The results of RFLP mapping will be useful for the selection and pyramiding of bacterial blight resistant genes.

Rediscovery of haploid breeding in the genomics era (유전체 시대에 반수체 육종의 재발견)

  • Lee, Seulki;Kim, Jung Sun;Kang, Sang-Ho;Sohn, Seong-Han;Won, So Youn
    • Journal of Plant Biotechnology
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    • v.43 no.1
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    • pp.12-20
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    • 2016
  • Advances in DNA sequencing technologies have contributed to revolutionary understanding of many fundamental biological processes. With unprecedented cost-effective and high-throughput sequencing, a single laboratory can afford to de novo sequence the whole genome for species of interest. In addition, population genetic studies have been remarkably accelerated by numerous molecular markers identified from unbiased genome-wide sequences of population samples. As sequencing technologies have evolved very rapidly, acquiring appropriate individual plants or populations is a major bottleneck in plant research considering the complex nature of plant genome, such as heterozygosity, repetitiveness, and polyploidy. This challenge could be overcome by the old but effective method known as haploid induction. Haploid plants containing half of their sporophytic chromosomes can be rapidly generated mainly by culturing gametophytic cells such as ovules or pollens. Subsequent chromosome doubling in haploid plants can generate stable doubled haploid (DH) with perfect homozygosity. Here, classical methodology to generate and identify haploid plants or DH are summarized. In addition, haploid induction by epigenetic regulation of centromeric histone is explained. Furthermore, the utilization of haploid plant in the genomics era is discussed in the aspect of genome sequencing project and population genetic studies.

Production of doubled haploid population derived from the microspore culture of rapeseed (Brassica napus L.) F1 generation and analysis of fatty acid composition (유채 잡종 1세대의 소포자 배양에 의한 배가반수체 집단 선발 및 지방산 조성 분석)

  • Lee, Ji Eun;Park, Ju Hyun;Kim, Kwang Soo;An, Da Hee;Cha, Young Lok
    • Journal of Plant Biotechnology
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    • v.49 no.1
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    • pp.74-81
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    • 2022
  • Brassica napus, an oil crop that produces rapeseed oil, is an allotetraploid (AACC, 2n = 38) produced by natural hybridization between B. rapa and B. oleracea. In this study, microspore was cultured using the F1 developed from a cross between 'EMS26' line with high oleic acid content and 'J8634-B-30' lines. The flower bud size showing the nuclear development at the late uninucleate and binucleate stage with high embryogenesis rate was 2.6 ~ 3.5 mm. Microspores were cultured using only this size and after then most microspore embryo developed into secondary embryos and then regeneration plants obtained from the developed multilobe. The analysis of the ploidy of the plants revealed that 66.7% and 27.8% of the total lines were tetraploids and octoploids, respectively. The sizes of stomatal cells in tetraploids, octoploids, and diploids were 25.5, 35.6, and 19.9 ㎛, respectively, indicating that ploidy level was positively correlated with cell size. Furthermore, 62 tetraploid doubled haploid (DH) lines were selected. The average oleic acid (C18:1) and linolenic acid (C18:3) concentrations of DH were 72.3% and 6.2%, respectively. Oleic acid and linolenic acid concentrations exceeded the two parental values in 5 and 14 DH lines, respectively, suggesting that these two fatty acids had transgressive segregation. Therefore, the DH population can be utilized for the biosynthesis of unsaturated fatty acids in rapeseed and related genes. It can also be used as a breeding material for varieties with high oleic acid concentrations.