• Title/Summary/Keyword: backcross

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Single Nucleotide Polymorphism Marker Discovery from Transcriptome Sequencing for Marker-assisted Backcrossing in Capsicum

  • Kang, Jin-Ho;Yang, Hee-Bum;Jeong, Hyeon-Seok;Choe, Phillip;Kwon, Jin-Kyung;Kang, Byoung-Cheorl
    • Horticultural Science & Technology
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    • v.32 no.4
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    • pp.535-543
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    • 2014
  • Backcross breeding is the method most commonly used to introgress new traits into elite lines. Conventional backcross breeding requires at least 4-5 generations to recover the genomic background of the recurrent parent. Marker-assisted backcrossing (MABC) represents a new breeding approach that can substantially reduce breeding time and cost. For successful MABC, highly polymorphic markers with known positions in each chromosome are essential. Single nucleotide polymorphism (SNP) markers have many advantages over other marker systems for MABC due to their high abundance and amenability to genotyping automation. To facilitate MABC in hot pepper (Capsicum annuum), we utilized expressed sequence tags (ESTs) to develop SNP markers in this study. For SNP identification, we used Bukang $F_1$-hybrid pepper ESTs to prepare a reference sequence through de novo assembly. We performed large-scale transcriptome sequencing of eight accessions using the Illumina Genome Analyzer (IGA) IIx platform by Solexa, which generated small sequence fragments of about 90-100 bp. By aligning each contig to the reference sequence, 58,151 SNPs were identified. After filtering for polymorphism, segregation ratio, and lack of proximity to other SNPS or exon/intron boundaries, a total of 1,910 putative SNPs were chosen and positioned to a pepper linkage map. We further selected 412 SNPs evenly distributed on each chromosome and primers were designed for high throughput SNP assays and tested using a genetic diversity panel of 27 Capsicum accessions. The SNP markers clearly distinguished each accession. These results suggest that the SNP marker set developed in this study will be valuable for MABC, genetic mapping, and comparative genome analysis.

Hybridization between Marine Medaka Oryzias dancena and Javanese Medaka Oryzias javanicus (바다송사리 Oryzias dancena와 자바송사리 Oryzias javanicus 간 잡종 유도)

  • Song, Ha-Yeun;Nam, Yoon-Kwon;Bang, In-Chul;Kim, Dong-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.43 no.5
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    • pp.462-473
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    • 2010
  • Inductions of hybrids and reciprocal hybrids between Oryzias dancena and O. javanicus (ODJ and OJD) were conducted and backcross hybrids between female O. dancena and male ODJ were also produced for biological and cytogenetic analysis. Embryonic development of ODJ and OJD were compared with those of their parents. Developmental time was fastest in O. dancena and ODJ, followed by O. javanicus and OJD. Oryzias dancena hatched 11 days (d) after fertilization, ODJ at 13 d, O. javanicus at 14 d and OJD at 15 d. The abnormality of external morphology rate in ODJ was 10.6%; however, OJD showed a high degree of abnormality, over 90%. The proportion of males was 90.0% and 31.3% for ODJ and OJD, respectively. Cytogenetic analysis was conducted to obtain basic information for genetic identification of O. dancena, O. javanicus and their hybrids. The karyotypes of all experimental groups showed 2n=48 chromosomes and the fundamental number (FN) was 48. The first pair carried secondary constrictions near the centromeric regions. Erythrocyte area and volume were $9.8\;{\pm}\;0.5\;{\mu}m^2$ and $18.2\;{\pm}\;1.0\;{\mu}m^3$, respectively, for O. dancena, $8.3\;{\pm}\;0.5\;{\mu}m^2$ and $15.8\;{\pm}\;1.5\;{\mu}m^3$ in O. javanicus, and $18.3\;{\pm}\;0.5\;{\mu}m^2$ and $15.7\;{\pm}\;1.3\;{\mu}m^3$ in ODJ. Erythrocyte area and volume in ODJ were similar to those of O. javanicus. In backcross hybrids between female O. dancena and male ODJ, all embryos failed to develop and died in the late gastrula stage.

QTL Mapping of Agronomic Traits in an Advanced Backcross Population from a Cross between Oryza sativa L. cv. Milyang 23 and O. glaberrima

  • Kang, Ju-Won;Suh, Jung-Pil;Kim, Dong-Min;Oh, Chang-Sik;Oh, Ji-Min;Ahn, Sang-Nag
    • Korean Journal of Breeding Science
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    • v.40 no.3
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    • pp.243-249
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    • 2008
  • In the previous study, 141 $BC_3F_2$ lines from a cross between the Oryza sativa cv. Milyang 23 and O. glaberrima were used to identify favorable wild QTL alleles for yield component traits. In this study, we carried out QTL analysis of four grain morphology as well as four yield component traits using 141 $BC_3F_5$ lines from the same cross and compared QTLs detected in two different generations. The mean number of O. glaberrima segments in the 141 $BC_3F_5$ lines ranged from 1 to 13 with 2.69 and 5.71 of the average means of homozygous and heterozygous segments, respectively. There was a three-fold difference in the number of QTLs detected for four traits commonly evaluated in two generations (seven QTLs in the $BC_3F_5$ vs 21 in the $BC_3F_2$ population). The percentages of the phenotypic variance explained by QTLs in the BC3F5 population were similar to or less than those in the $BC_3F_2$ population. This is probably due to the difference in the genetic composition of two populations and the environmental effects. The locations of the QTLs commonly detected in both generations were in good agreement except for one QTL for spikelets per panicle. The yield QTL, yd3 was colocalized with the spikelets per panicle, spp3. Yield increase at this locus is due to the increase in spikelets per panicle, because both traits were associated with increase in spikelets per panicle and yield due to the presence of an O. glaberrima allele. Clusters of QTLs for grain morphology traits were observed in two chromosome regions. One cluster harboring five QTLs near SSR markers RM106 and RM263 was detected on chromosome 2. This population would serve as a foundation for development of the introgression line population from a cross between Milyang 23 and O. glaberrima.

Identification of a Rice Gene (Bph 1) Conferring Resistance to Brown Planthopper (Nilaparvata lugens Stal) Using STS Markers

  • Kim, Suk-Man;Sohn, Jae-Keun
    • Molecules and Cells
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    • v.20 no.1
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    • pp.30-34
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    • 2005
  • This study was carried out to identify a high-resolution marker for a gene conferring resistance to brown planthopper (BPH) biotype 1, using japonica type resistant lines. Bulked segregant analyses were conducted using 520 RAPD primers to identify RAPD fragments linked to the BPH resistance gene. Eleven RAPDs were shown to be polymorphic amplicons between resistant and susceptible progeny. One of these primers, OPE 18, which amplified a 923 bp band tightly linked to resistance, was converted into a sequence-tagged-site (STS) marker. The STS marker, BpE18-3, was easily detectable as a dominant band with tight linkage (3.9cM) to Bph1. It promises to be useful as a marker for assisted selection of resistant progeny in backcross breeding programs to introgress the resistance gene into elite japonica cultivars.

THE EFFECTS OF POPULATION SIZE AND DOMINANCE OF QUANTITATIVE TRAIT LOCI (QTL) ON THE DETECTION OF LINKAGE BETWEEN MARKERS AND QTL FOR LIVESTOCK

  • Jeon, G.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.8 no.6
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    • pp.651-655
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    • 1995
  • A simulation study on detection of linkage between genetic markers and QTL in backcross design was conducted. The effects of various sample sizes and the degree of QTL dominance on detention of linkage were examined by using a simple regression analysis. The results indicated that as sample size increased, the standard error of the estimated slope became smaller. When the dominance effect of QTL was complete, the estimated slope tended to be negative but was statistically not significant at all with type I error of greater than 50%. With complete linkage between genetic Marker and QTL, the estimated intercept value was smallest but the estimated slope was largest as expected. In most cases with various degree of dominance and sample sizes, when the actual recombination rate became larger, greater values were obtained for the slope except in the case of complete dominance of QTL.

Breeding of Bivoltine Breeds of Bombyx mori L Suitable for Variable Climatic Conditions of the Tropics

  • Moorthy, S. M.;Das, S. K.;Kar, N. B.;Urs, S. Raje
    • International Journal of Industrial Entomology and Biomaterials
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    • v.14 no.2
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    • pp.99-105
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    • 2007
  • The success of rearing with presently available conventional bivoltine is unpredictable in some seasons of the tropical regions due to highly fluctuating adverse climatic conditions. Thus, in order to popularize bivoltine breeds in tropical parts of India, it is very much essential to have a bivoltine breed(s), which can give stable cocoon crop under variable environments. With this objective a breeding programme was undertaken to improve the survival trait in bivoltine silkworm by introducing multivoltine genes into bivoltine through back crossing. Resultant bivoltine lines showed significantly higher survival in compared to the receptor (Bivoltine) parent and control bivoltine breed. Esterase isozyme analysis revealed similar banding pattern in the developed bivoltine and in the donor multivoltine, which predicts the introgression of multivoltine character into evolved bivoltine.

Evaluation of Germplasm and Development of SSR Markers for Marker-assisted Backcross in Tomato (분자마커 이용 여교잡 육종을 위한 토마토 유전자원 평가 및 SSR 마커 개발)

  • Hwang, Ji-Hyun;Kim, Hyuk-Jun;Chae, Young;Choi, Hak-Soon;Kim, Myung-Kwon;Park, Young-Hoon
    • Horticultural Science & Technology
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    • v.30 no.5
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    • pp.557-567
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    • 2012
  • This study was conducted to achieve basal information for the development of tomato cultivars with disease resistances through marker-assisted backcross (MAB). Ten inbred lines with TYLCV, late blight, bacterial wilt, or powdery mildew resistance and four adapted inbred lines with superior horticultural traits were collected, which can be useful as the donor parents and recurrent parents in MAB, respectively. Inbred lines collected were evaluated by molecular markers and bioassay for confirming their disease resistances. To develop DNA markers for selecting recurrent parent genome (background selection) in MAB, a total of 108 simple sequence repeat (SSR) primer sets (nine per chromosome at average) were selected from the tomato reference genetic maps posted on SOL Genomics Network. Genetic similarity and relationships among the inbred lines were assessed using a total of 303 polymorphic SSR markers. Similarity coefficient ranged from 0.33 to 0.80; the highest similarity coefficient (0.80) was found between bacterial wilt-resistant donor lines '10BA333' and '10BA424', and the lowest (0.33) between a late blight resistant-wild species L3708 (S. pimpinelliforium L.) and '10BA424'. UPGMA analysis grouped the inbred lines into three clusters based on the similarity coefficient 0.58. Most of the donor lines of the same resistance were closely related, indicating the possibility that these lines were developed using a common resistance source. Parent combinations (donor parent ${\times}$ recurrent parent) showing appropriate levels of genetic distance and SSR marker polymorphism for MAB were selected based on the dendrogram. These combinations included 'TYR1' ${\times}$ 'RPL1' for TYLCV, '10BA333' or '10BA424' ${\times}$ 'RPL2' for bacterial wilt, and 'KNU12' ${\times}$ 'AV107-4' or 'RPL2' for powdery mildew. For late blight, the wild species resistant line 'L3708' was distantly related to all recurrent parental lines, and a suitable parent combination for MAB was 'L3708' ${\times}$ 'AV107-4', which showed a similarity coefficient of 0.41 and 45 polymorphic SSR markers.

Marker Assisted Selection of Brown Planthopper Resistance and Development of Multi-Resistance to Insect and Diseases in Rice (Oryza sativa L.) (DNA 마커를 이용한 벼멸구 저항성 선발 및 복합내병충성 벼 계통 육성)

  • Lee, Jong-Hee;Yeo, Un-Sang;Cho, Jun-Hyun;Lee, Ji-Yoon;Song, You-Chun;Shin, Mun-Sik;Kang, Hang-Won;Sohn, Jae-Keun
    • Korean Journal of Breeding Science
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    • v.43 no.5
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    • pp.413-421
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    • 2011
  • The main objective of this study was to develop the multi-resistance lines to insects(brown planthopper; BPH, rice green leafhopper; GRH) and disease(blast; BL, bacterial blight; BB and rice stripe virus disease;RSV) with good grain quality and plant type by combining conventional breeding and marker assisted selection(MAS) and to eliminate the linkage drag effects between Bph1 gene and culm length, we conducted MAS of Bph1 gene in advanced backcross and double cross progenies. 'Nampyeong', 'Junam' and 'Milyang220' were used as the parent in this study. 'Milyang220' was used as the donor of brown planthopper resistance gene Bph1 with tall culm length. Two backcross progenies were developed using two recipients 'Nampyeong' carrying GRH resistance gene Grh3(t) with good grain appearance and 'Junam' harboring bacterial blight resistance gene Xa3 with short culm length. Two $BC_1$ generations were resulted from the backcrossing of the $F_1$ plants with recurrent parents 'Nampyeong' and 'Junam'. The second rounds of backcrossing($BC_2$) were derived from the cross of selected resistant $BC_1F_1$ plants based on heterozygous genotype of RM28493 linked to Bph1 gene. The double crossed population was constructed from the cross of between each heterozygous $BC_2F_1$ plants at RM28493 locus of '$Nampyeong^*3$ / Milyang220' and '$Junam^*3$ / Milyang220'., The homozygous alleles in Bph1 gene were selected using co-dominant DNA marker RM28493 in double crossed population. Eighty-five lines with multi-resistance to BL, BB, RSV, GRH and BPH were selected by bio-assay and MAS in generation of double crossing. The culm length, head rice ratio and yield of the selected multi resistance lines was ranged from 71 to 88 cm, from 51 to 93%, from 449 to 629 kg/10a. respectively. We can select a promising multi resistance line similar with 'Nampyeong' of major agronomic traits such as culm legnth, head rice ratio and yield. It was designated as Milyang265. Finally this study was developed the multi resistant varieties against to insects and diseases with the good grain quality 'Milyang265' by the advanced backcross and double cross combining MAS and it can be used as genetic resources of multi-resistance to insect and diseases in rice breeding programs.