• Journal of Plant Biotechnology
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• v.5 no.1
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• pp.51-57
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• 2003

Doubled haploids have long been recognized as a valuable tool in plant breeding since it not only offers the quickest method of advancing heterozygous breeding lines to homozygosity, but also increased the selection efficiency over conventional procedures due to better discrimination between genotypes within any one generation. Salt tolerant mutants were obtained in rice the variety, 'Hawsungbyeo', through in vitro mutagenesis of in vitro cultured anther-derived calli. Various doses (30, 50, 70 and 90 Gy) of gamma ray were applied to investigate the effect of radiation on callus formation on medium containing 1% NaCl, green plant regeneration, frequency of selected doubled haploid mutants and of the salt tolerant screen. It was demonstrated that the dose of 30 and 50 Gy gamma rays had significant effects on callus formation, regeneration and selection of salt tolerance. No tolerant lines were obtained from non-mutagenized cultures. From gamma ray irradiated cultures, five tolerant lines ($M_2$generation) at germination stage and 13 tolerant lines ($M_3$genoration) at seedling stage were obtained. The frequency of salt tolerant mutants indicates that anther culture applied in connection with gamma rays is an effective way to improve salt tolerance.

• Journal of Plant Biotechnology
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• v.35 no.4
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• pp.257-263
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• 2008

Improvement of the macro- and micro-elements density of rice (Oryza sativa L.) is gradually becoming a new breeding objective. In this study, the genomic regions associated with potassium, calcium, magnesium and iron content in rice grain were identified and characterized by using a doubled haploid (DH) population. Fifty-six simple sequence repeat (SSR) and one hundred and twelve sequence tagged site (STS) markers were selected to construct the genetic linkage map of the DH population with a full length of 1808.3cM scanning 12 rice chromosomes. Quantitative trait loci (QTLs) were detected, and QTL effects and QTL interactions were calculated for five traits related to macro- and micro-elements in the DH population from a cross between 'Samgang' (Tongil) and 'Nagdong' (Japonica). Twelve QTLs were located on five chromosomes, consisting of two QTLs for potassium, three QTLs for calcium, two QTLs for magnesium, one QTL for iron content and four QTLs for the ratio of magnesium to potassium (Mg/K). Among them, qca1.1 was detected on chromosome 1 with an LOD value of 8.58 for calcium content. It explained 27% of phenotype variations with increasing effects from 'Samgang' allele. Furthermore, fifteen epistatic combinations with significant interactions were observed on ten chromosomes for five traits, which totally accounted for 4.19% to 12.72% of phenotype variations. The screening of relatively accurate QTLs will contribute to increase the efficiency of marker-assisted selection (MAS), and to accelerate the establishment of near-isogenic lines (NILs) and QTL pyramiding.

• Plant Biotechnology Reports
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• v.3 no.4
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• pp.317-324
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• 2009

The insecticidal toxin gene of Bacillus thuringiensis (Bt) is one of the most commonly used in the development of genetically modified (GM) crops. In this research, we analyzed Bt rice showing lepidopteran pest-resistance. The Bt gene is a synthetic Cry1Ac composed of optimal codons for plants, and the Bt protein is targeted to the chloroplast by a transit peptide. Three Cry1Ac rice events (C103-3, C127-1, and C7-1) were analyzed for molecular characterization. C103-3 contains two copies of T-DNA where the left border (LB) region is truncated. Both C7-1 and C127-1 have a single copy of T-DNA, but a part of the vector backbone DNA is inserted into the genome of C127-1; thus, only C7-1 had intact T-DNA. Progenies of C7-1 crossed with the original cultivar, Nakdong, and double-haploid lines from anther culture of lines crossed with the elite cultivar, Dongjin, were analyzed for T-DNA flanking genomic DNA and genotyping. Results showed that an intact T-DNA region without the vector backbone was inserted into the genome and was stably inherited through generations. The C7-1 homozygous event could be used as breeding material to develop GM rice with pest resistance.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.55 no.2
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• pp.105-110
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• 2010

This study was conducted to analyze the relationships between resistance of brown planthopper and traits related to the lodging in rice. For the linkage analysis of traits tested in this study, a genetic linkage map was created with 162 DNA markers spanning 12 rice chromosomes based on 120 doubled haploid (DH) lines, which were derived from a cross between Samgang', a Tongil type cultivar with BPH resistance, and ‘Nagdong’, a japonica cultivar. QTLs were identified to analyze the agronomic traits including lodging by composite interval mapping. Thirteen QTLs were detected for five traits comprised of plant length (PL), 3rd internode length (3rdIL), moments (Mo), lodging index (LI), and breaking weight (BW). The relationships between the BPH resistance and agronomic traits including lodging revealed that two QTLs (qBPR7, qBPR8) were linked to traits related to lodging. Two QTLs, qBPR7 and qBPR8 on chromosome 7 (RM531-7042) and 8 (RM1148- RM544) showed associations with moments and 3rd internode length, respectively.

• Journal of Plant Biotechnology
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• v.37 no.2
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• pp.125-132
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• 2010

Rice is the staple food of more than 50% of the worlds population. Cultivated rice has the AA genome (diploid, 2n=24) and small genome size of only 430 megabase (haploid genome). As the sequencing of rice genome was completed by the International Rice Genome Sequencing Project (IRGSP), many researchers in the world have been working to explore the gene function on rice genome. Insertional mutagenesis has been a powerful strategy for assessing gene function. In maize, well characterized transposable elements have traditionally been used to clone genes for which only phenotypic information is available. In rice endogenous mobile elements such as MITE and Tos (Hirochika. 1997) have been used to generate gene-tagged populations. To date T-DNA and maize transposable element systems has been utilized as main insertional mutagens in rice. A main drawback of a T-DNA scheme is that Agrobacteria-mediated transformation in rice requires extensive facilities, time, and labor. In contrast, the Ac/Ds system offers the advantage of generating new mutants by secondary transposition from a single tagged gene. Revertants can be utilized to correlate phenotype with genotype. To enhance the efficiency of gene detection, advanced gene-tagging systems (i.e. activation, gene or enhancer trap) have been employed for functional genomic studies in rice. Internationally, there have been many projects to develop large scales of insertionally mutagenized populations and databases of insertion sites has been established. Ultimate goals of these projects are to supply genetic materials and informations essential for functional analysis of rice genes and for breeding using agronomically important genes. In this report, we summarize the current status of Ac/Ds-mediated gene tagging systems that has been launched by collaborative works from 2001 in Korea.

• Korean Journal of Plant Tissue Culture
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• v.24 no.6
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• pp.319-322
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• 1997

The optimal conditions for callus formation and plant regeneration were determined by the manipulation of culture method in unpollinated ovary culture of rice. The effect of cold pretreatment on callus formation and plant regeneration varied with duration of pretreatment. The maximum frequency (38.7%) of plant regeneration was obtained from the unpollinated ovary pretreated for 10 days at $12^{\circ}C$. The ability of callus formation and plant regeneration was higher on the medium with picloram (1 mg/L) than that of 2, 4-D (1 mg/L). However, the high concentration of picloram increased markedly the frequency of albino plant from unpollinated ovary-derived callus. Floral parts inoculated as a unit play important roles in callus formation and plant regeneration. Best result was obtained when ovary with partial cut glume, pedicel and secondary branch as a unit was cultured.

• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.13-13
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• 2022

Recently, stable crop production is threatened by the effects of climate change. In particular, it is difficult to consistently maintain agricultural policies due to large price fluctuations depending on the difference in total domestic rice production from year to year. For stable rice production amid changes in the crop growing environment, development of varieties with improved disease resistance and abiotic stress stability is becoming more important. In here, drought and cold tolerant trait have been studied. First, for the development of drought tolerant varieties, we analyzed which agricultural traits are mainly affected by domestic drought conditions. As a result, it was observed that drought caused by the lack of water during transplanting season inhibits the development of the number of tiller and reduces the yield. 'Samgang' was selected as a useful genetic resource with strong drought tolerant and stable tiller number development even under drought conditions by phenotype screening. Three of drought tolerant QTLs were identified using doubled haploid (DH) population derived from a cross between Nacdong and Samgang, a drought sensitive and a tolerant, respectively. Among these QTLs, when qVDT2 and qVDTl1 were integrated, it was investigated that the tiller number development was relatively stable in the rainfed paddy field conditions. It is known that the high-yielding Tongil-type cultivars are severely affected by cold stress throughout the entire growth stage. In this study, we established conditions that can test the cold tolerance phenotype with alternate temperature to treat low temperatures in indoor growth conditions similar to those in field conditions at seedling stage. Three cold tolerant QTLs were explored using population derived from a cross between Hanareum2 (cold sensitive variety, Tongil-type) and Unkwang (cold tolerant variety, Japonica). Among these QTLs, qSCT12 showed strong cold tolerant phenotype, and when all of three QTLs were integrated, it was investigated that cold tolerant score was relatively similar to its donor parent, Unkwang, in our experimental conditions. We are performing that development of new variety with improved cold tolerant through the introduction of these QTLs.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.64 no.1
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• pp.1-17
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• 2019

A quantitative trait loci (QTL) analysis of traits related to heading and yield was performed develop rice cultivars that are both early maturing and panicle weight type varieties. Our analysis included 120 strains of the Cheongcheong Nagdong doubled haploid (CNDH) variety. An observational growth experiment was conducted to identify genetic agronomic traits of CNDH. Heading date, ten plant weight, moisture, thousand grain weight, and yield had a normal distribution based on the frequency distribution table of the observational growth data. The QTL analysis found one heading-related and nine yield-related QTLs. The LOD of 2.85 was the largest in QTLs for heading date (QHD), 5.39 in QTLs for ten plant weight (QTPW), 3.92 in QTLs for moisture (QM), 4.80 in QTLs for thousand grain weight (QTGW), and 3.7 in QTLs for yield (QY). Genomic analysis detected 58 candidate genes on chromosome 2, 3, 7, 8, and 10. Among those, we found Rcd1 protein and OsERF3 gene in QM, MtN3 and zinc finger protein genes in QTGW, and OsNAC3 protein gene in QY. If further analysis reveals the presence of genes related to water content, thousand grain weight or yield in the CNDH stains, we can develop a selection of varieties that will be capable of coping with climate change and will contribute to global food problems.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.67-67
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• 2017

We developed the breeding materials with diverse grain size and shape in japonica rice. Grain size and shape are important factors affecting consumer preference and choice. However, most of Korean japonica rice cultivars have small, short, and round grain. To diversify the grain size and shape of japonica rice, we conducted the breeding program using donor parents, Jizi1560 and Jizi1581. Jizi1560 and Jizi1581 are japonica germplasm with extremely large grain. Four crosses between the each donor parents and high yielding japonica rice cultivars, Deuraechan and Boramchan, were constructed and then anther culture method was applied. We obtained 290 doubled-haploid (DH) lines with appropriated morphological traits and selected 91 DH lines with diverse grain size and shape. The grain related-traits of the selected DH lines showed a higher diversity when compared with 319 cultivars developed by NICS (264 japonica, 13 black, and 32 Tongil type cultivars). We designated the selected DH lines, four parents, and Daeripbyeo 1, large grain japonica cultivar, as the breeding materials for further analysis. The breeding materials were classified into five groups, A to E, based on the grain-related traits. Group A (including Jizi1581) and Group B (including Daeripbyeo 1) showed similar grain width, whereas Group A exhibited longer grain length and heavier grain weight. Group C (including Deuraechan and Boramchan) showed shorter and rounder grain shape and smaller grain size than any other groups. Group D including solely Jizi1560 had extremely large grain, such as the longest grain length, width, and thickness and heaviest grain weight. Group E including only two DH lines had long and slender grain shape, so that showed the highest ratio of length to width. The grain size and shape of the breeding materials exhibited beyond the characteristics of previously developed Korean japonica cultivars. The breeding materials will be applied in the breeding programs to diversify the grain size and shape of japonica rice.

• Korean Journal of Breeding Science
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• v.41 no.3
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• pp.236-243
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• 2009

This study was conducted to develop a japonica-type rice cultivar with brown planthopper (BPH) resistance using DNA markers. A doubled haploid (DH) population consisting of 120 pure-lines was established by anther culture of $F_1$ hybrids between 'Samgang', a Tongil type BPH resistance cultivar, and 'Nagdong', a japonica cultivar. To determine the map position of genes responsible for BPH resistance in rice, a genetic map was constructed based on 120 DH lines. A total of 162 molecular markers were classified into 12 linkage groups, covering 1,884 Kosami centimorgan (cM) with an average of 11.6 cM. Five QTLs (qBPR3, qBPR6, qBPR7, qBPR8, and qBPR12) associated with BPH resistance were identified and mapped on chromosomes 3, 6, 7, 8, and 12, respectively, using the genetic map constructed in this study. To analyze the relationship between BPH resistance and agronomic traits, a total of eight QTLs related to the agronomic traits were detected on 12 rice chromosomes. In an analysis of relationships, three QTLs (qBPR3, qBPR7, and qBPR8) showed a linkage with tested agronomic traits. A QTL (qBPR3) located on chromosome 3 (RM282-3023) was closely linked to culm length (qCL3). The QTL (qBPR8) for BPH resistance on the short arm of chromosome 8 also overlapped the region detected in culm length (qCL8).