To diversify the grain quality characteristics of Korean varieties of japonica rice (Oryza sativa japonica), recombinant inbred lines were developed from a cross between the Korean cultivar “Boramchan” and the tropical japonica cultivar “Pecos” from the United States. Quantitative trait locus (QTL) analysis was performed to investigate the traits related to grain quality. Sixteen traits were evaluated, including five related to grain appearance, six associated with pasting properties and glossiness of cooked rice, and four related to eating texture. QTLs were detected on chromosomes 3, 5, 6, and 10, and candidate genes, including Hd1, qLTG3-1, GW5, and qGS10 were identified. The heading date-related gene Hd1 was associated with the following traits: head rice (HR), opaque rice (OR), damaged rice, pasting temperature, peak viscosity (PV), breakdown viscosity (BD), and glossiness. Most quality-related traits were influenced by allelic variation at Hd1, and the Hd1⁺ allele from “Boramchan”, which delays heading, positively affected both grain appearance and eating quality. The low-temperature germinability gene qLTG3-1 was identified as a candidate gene for broken rice and several pasting properties, including trough, final, and breakdown viscosities, and was shown to influence pasting characteristics depending on allelic variation. The grain shape gene GW5 was a candidate gene for HR and whiteness, whereas qGS10 was associated with OR, PV, BD, and setback viscosity. The GW5⁺ and qGS10^P alleles, derived from the slender grain shape of “Pecos,” contributed positively to both grain appearance and eating quality, indicating their potential as favorable alleles for improving grain shape and diversifying quality traits in Korean japonica rice. The results of this study demonstrate that key agronomic trait-related genes, such as heading and grain shape, significantly affect various quality-related traits, highlighting the potential of introducing and combining novel alleles to enhance grain quality in Korean japonica rice.

Leaf size is closely related to photosynthesis and greatly affects rice productivity. Therefore, the search for quantitative trait Locus (QTL) that regulates leaf size is important for improving productivity through rice breeding. The F₉ recombinant inbred line (RIL) population from the cross between ‘Unbong 40’ (large leaves) and ‘Odae’ (ordinary size leaves) was grown in a test field, and the size traits of the flag leaves and 2nd leaves, along with stem and panicle length, were measured. Through QTL analysis of these traits, we detected 20 QTLs on chromosomes 1, 3, 6, 9, 11, and 12. In particular, the flag leaf length QTL qFLL3, the flag leaf area QTL qFLA3, the 2nd leaf length QTL q2LL3-2, and the 2nd leaf area QTL q2LA3 were clustered in the 149.0-161.6 cM region of chromosome 3, indicating that one QTL gene in this region may have pleiotropic effects that regulate the flag leaf length, flag leaf area, 2nd leaf length, and 2nd leaf area. In addition, the flag leaf width QTL qFLW9 and 2nd leaf width QTL q2LW9 were clustered in the 2.1-22.86 cM region on chromosome 9, which suggests that one QTL gene in this region may have pleiotropic effects that regulate both the flag leaf width and 2nd leaf width. These results serve as a valuable reference for breeding programs aiming to increase photosynthesis and yield by identifying QTLs for leaf size traits in Korean japonica rice.

The rice cultivar ‘Chamdongjin’ was developed to enhance the disease resistance of ‘Sindongjin’. ‘Sindongjin’, developed by the RDA in 1999, is a mega-variety with a unique grain size and excellent taste, and has been cultivated in the largest area in Korea since 2018. As ‘Sindongjin’ has been cultivated in a large area recently, problems such as pest damage are increasing, and the necessity to develop alternative cultivars is emerging. Accordingly, the multiple disease-resistant, mid-late maturing rice cultivar ‘Chamdongjin’, carrying the bacterial blight resistance gene Xa21 into the genetic background of ‘Sindongjin’, was developed. ‘Chamdongjin’ was developed by the backcrossing method using the donor parent ‘HR27195-59-3-5-5’, harboring the bacterial blight resistance genes Xa3+Xa21 and the recurrent parent ‘Sindongjin’. ‘Chamdongjin’ was selected through the pedigree method, yield trials, and local adaptability tests, with a high selection pressure for agronomic trait similarities with ‘Sindongjin’. In order to effectively improve the disease resistance, marker-assisted selection for disease resistance genes and bioassays for bacterial blight, blast, and stripe virus were performed in parallel. ‘Chamdongjin’ was confirmed to have three disease-resistance genes, Xa3+Xa21+Stvb-i. Most of agronomic traits of ‘Chamdongjin’ were similar to ‘Sindongjin’, while ‘Chamdongjin’ showed enhanced disease resistance against bacterial blight and panicle blast compared to ‘Sindongjin’. ‘Chamdongjin’, like ‘Sindongjin’, possessed a grain size-related gene, the gs3 allele, and exhibited larger grains than general Korean japonica cultivars. In addition, ‘Chamdongjin’ showed a similar or higher level of palatability compared to ‘Sindognjin’, indicating the excellent quality characteristics. In the genetic background analysis using 787 KASP markers, ‘Chamdongjin’ showed 96.3% homozygosity with ‘Sindongjin’, indicating that it had a more similar genetic background to ‘Sindongjin’ than to other existing cultivars. ‘Chamgdongjin’ is expected to be widely cultivated as an alternative to ‘Sindongjin’, which could maintain the brand value while preventing the disease damage caused to ‘Sindongjin’ (Registration No. 9310).

Many studies concerning breeding maize varieties are in progress in Korea and other countries. Double haploid technology is widely used for the development of commercial maize varieties worldwide, and has also been utilized in Korea since its introduction by the Maize Research Institute, Gangwondo. We performed a study to improve the efficiency of selecting haploid maize seeds using fluorescence imaging. It was verified that anthocyanin produced by the expression of R1-nj gene can be detected by fluorescence imaging, and we developed a high-throughput method for discriminating between haploid and diploid seeds. Compared with discriminating with naked eye, this method reduced the time for discriminating haploid and diploid maize by 91.7% and increased selection accuracy by 16.8% for haploid and 2.2% for diploid maize. This method enabled the acquisition of more haploid seeds and improved the efficiency of breeding research by shortening the time involved.

Fast and accurate selection is essential for breeding to cope with rapid climate changes and a steeply increasing population. Consequently, technologies for high-throughput phenotyping (HTP) are emerging. These technologies, unlike conventional phenotyping methods, enable us to evaluate agronomic traits in a fast and massive manner. Thus, the HTP facility was built to acquire and analyze crop images using RGB sensors at the National Institute of Agricultural Sciences, Republic of Korea. By testing various conditions to acquire images, we determined the conditions for phenotyping using the RGB sensor as follows: exposure 30,000 ms, gamma 75, and gain 100 using LED lights in a blue background. Based on this condition, images from 96 individual plants of rice Dongjin cultivar were obtained every week to measure plant height and shoot area, which are directly associated with yield. The results obtained from the image analysis were compared with the manually collected results. The r² value between the projected plant height obtained from image analysis and the plant height obtained from manual measurement was 0.989. Furthermore, the r² value between the projected shoot area obtained from image analysis and the shoot area obtained from manual measurement was 0.981. These results show that image analysis is highly reliable and can be used for crop phenotyping. Therefore, we expect that the new method we developed will be used for breeding in the near future.

The cultivation of early maturing rice in the plain areas in Korea is increasingly diversifying its cropping systems. Early maturing rice cultivars are usually vulnerable to bacterial blight (BB) and rice stripe virus (RSV) compared to mid- and mid-late maturing rice cultivars. To enhance the resistance of early maturing rice against BB and RSV, ‘IS592BB’ was developed by introgression of resistance genes, Xa3, Xa21, and Stvb-i into the genetic background of ‘Unkwang’, an early maturing rice cultivar with excellent agronomic characteristics. ‘IS592BB’ was derived from a backcross between ‘Unkwang’ and the F1 cross between ‘Unkwang’ and ‘SR31206-12’ (‘Unkwang’/‘SR30075’) and selected through the pedigree method, yield trials, and local adaptability tests. Using bioassay and marker-assisted selection to BB and RSV, Xa3, Xa21, and Stvb-i were pyramided in ‘IS592BB’. ‘IS592BB’ exhibited high-level and broad-spectrum resistance against BB, including the K3a race, the most virulent race in Korea, and also showed a resistance reaction to RSV. ‘IS592BB’ was found to be an early maturing rice cultivar with similar agronomic characteristics to ‘Unkwang’ such as early maturing, lodging tolerance with short culm, erect plant architecture, blast resistance, and high-yielding performance. In the genetic background analysis using 771 KASP (Kompetitive Allele-Specific PCR) markers, ‘IS592BB’ was confirmed to be the near-isogenic line (NIL) of ‘Unkwang’ with a 93.6% recovery rate. ‘IS592BB’ was successfully introgressed with the Xa3+Xa21+Stvb-i genes into ‘Unkwang’ without linkage drag negatively affecting its agronomic characteristics. ‘IS592BB’ would enhance the adaptability of early maturing rice in the plain area through its multiple disease resistance (Registration No. 7648).

A dwarf mutant rice line was selected from an Ac/Ds insertion mutant population and named dwf1. The phenotype of F₁ and F₂ plants derived from a cross between dwf1 and Dongjin indicated that a single recessive gene is responsible for the mutant phenotype, and we named this gene dwf1. Resequencing of the dwf1 line and Dongjin (wild type) revealed 42,386 homozygous single nucleotide polymorphisms (SNPs) between dwf1 line and Dongjin. MutMap analysis was performed by sequencing a DNA pool prepared from 100 mutant type plants in the dwf1/Dongjin F₂ population, and it was found that the dwf1 gene was located in the 23 ~ 30 Mbp region on chromosome 4. In this region, we found a non-synonymous SNP in the Os04g0469800 gene, which was reported as D11 gene encoding a cytochrome P450 family protein involved in the biosynthesis of brassinosteroids (BRs). This SNP was regarded as the causative SNP for the dwf1 phenotype, and the dwf1 gene is a novel allele of D11. We performed mapping of the dwf1 gene with five SNP markers on chromosome 4 with 190 dwf1/Dongjin F₂ plants. The phenotype of F₂ plants was completely co-segregated with genotypes of the J10402 marker, which was developed based on the non-synonymous SNP in the D11 gene. These results will contribute to the study of the molecular biological functions of the D11 gene and BRs.

Perilla is an oilseed crop cultivated in Korea since ancient times. Due to the high α-linolenic acid content in perilla, perilla seed oil can easily become rancid. α-Linolenic acid is synthesized by two enzymes, endoplasmic reticulum-localized Δ15 desaturase (FAD3) and chloroplast-localized Δ15 desaturase (FAD7) in vivo. In order to lower the α-linolenic acid content of the seed oil without disturbing plant growth, we tried to suppress the expression of only the FAD3 gene using RNA interference, whilst maintaining the expression of the FAD7 gene. Seventeen transgenic plants with herbicide (Basta™) resistance were obtained by Agrobacterium-mediated transformation using hypocotyls of perilla plants. The transgenic plants were firstly confirmed by treatment with 0.3% (v/v) Basta™ herbicide, and the expression of FAD3 was measured by Northern blot analysis. The α-linolenic acid content was 10-20%, 30-40%, and 60% in two, seven, and three of the twelve T₁ transgenic perilla plants which had enough seeds to be analyzed for fatty acid composition, respectively. Analysis of the fatty acid composition of T₂ progeny seeds from T₁ plants with the lowest α-linolenic acid content showed that the homozygous lines had 6-10% α-linolenic acid content and the heterozygous lines had 20-26% α-linolenic acid content. It is expected that the reduction in α-linolenic acid content in perilla seed oil will prevent rancidity and can be utilized for the production of high-value functional ingredients such as high γ-linolenic acid.

Gayabyeo, a Tongil-type rice variety, has been known to be resistant to the brown planthopper (BPH) in Korea. For genetic analysis of BPH resistance of Gayabyeo, we developed an F2 and F3 population derived from a cross between Gayabyeo and Taebaegbyeo which is a Tongil-type BPH susceptible rice variety. Based on the previously detected 284,501 putative SNPs between Gayabyeo and Taebaegbyeo, 99 cleaved amplified polymorphic sequences (CAPS) markers were developed, and they have been used for genotyping 180 F2 plants. By comparison of resequencing data of Gayabyeo and the sequences of already reported BPH resistance genes (Bph3, BPH9, Bph14, BPH18, BPH26), it was revealed that Gayabyeo has Bph3 and BPH26 resistance genes. Two InDel markers, Bph3IND and BPH26IND, were developed, which can be used as selection markers in breeding program aiming at introducing BPH resistance genes of Gayabyeo into Korean high quality japonica rice varieties. In addition, BPH bioassay was performed with 180 F3 lines for BPH resistance QTL analysis. Two major QTLs were found on chromosome 4 and 12. The regions of these two QTLs included Bph3 and BPH26, which also supported that Gayabyeo has Bph3 and BPH26 resistance genes. These results would be useful in accelerating development of various BPH-resistant high quality japonica rice varieties in Korea.

As a first step of mapping genes conferring resistance to the brown planthopper, Nilaparvata lugens Stål, in Gayabyeo using a population derived from a cross between Gayabyeo and Taebaegbyeo, we performed the whole genome resequencing of these two Tongil-type rice varieties. The amount of raw sequence data was about 18.5X10⁹ bp and 17.9X10⁹ bp in Gayabyeo and Taebaegbyeo, respectively. After quality trimming and read mapping onto Nipponbare reference genome sequence, 9.3X10⁹ bp was mapped in Gayabyeo with mapping depth of 25.0X, and 9.5X10⁹ bp was mapped in Taebaegbyeo with mapping depth of 25.5X. Between Gayabyeo and Nipponbare, 1,585,880 SNPs were detected, while 1,416,898 SNPs were detected between Taebaegbyeo and Nipponbare. Between Gayabyeo and Taebaegbyeo, 284,501 SNPs were detected. Among the SNPs between Gayabyeo and Taebaegbyeo, 21.2% were in genic region and 78.8% were in intergenic region. In CDS region, 15,924 SNPs were detected, among which synonymous SNPs covered 47.3% and non-synonymous SNPs covered 52.7%. We designed Cleaved Amplified Polymorphic Sequences (CAPS) markers with SNPs in the restriction enzyme recognition sites, and 20 CAPS markers were tested. Of the 20 markers, 19 markers showed polymorphism and one marker showed monomorphism between Gayabyeo and Taebaegbyeo. It is expected that sufficient DNA markers for mapping genes with a population derived from a cross between Gayabyeo and Taebaegbyeo can be developed based on the results of the study.

The rice recombinant inbred lines derived from Milyang23 and Gihobyeo cross were used in genetic mapping and QTL analysis studies. In this study, we developed a new 101 CAPS markers based on the SNPs in the whole genome region between these varieties. As a result, the total genetic distance and average distances were 1,696.97 cM and 3.64 cM, respectively. In comparison to the distance of the previous genetic map constructed based on 365 DNA markers, the new genetic map was found to have a decreased distance. The map was applied for the detection of QTLs on all seven traits relevant to diameter of stem internode, length of culms, length of panicles and the number of panicles including the correlation analysis between each trait. The QTLs results were similar to the report in previous studies, whereas the distance between the markers was narrowed and accuracy increased with the addition of 101 CAPS markers. A total of 9 new QTLs were detected for stem internode traits. Among them, qI1D-6 had higher LOD of 5.1 and phenotype variation of 50.92%. In this experiment, a molecular map was constructed with CAPS markers using next generation sequencing showing high accuracy for markers and QTLs. In the future, developing more accurate QTL information on stem internode diameters with various agriculturally important traits will be possible for further rice breeding.

Crops are exposed to various environmental stresses. These have been affecting the growth of crops, resulting in the severe loss of agronomic production in many countries. Therefore, development of new varieties of resistant crops is required to assure the desired productivity of crops in stress conditions. In this study, a putatively stress-related gene BrTSR53 was isolated from Brassica rapa. The BrTSR53 is 481 bp long and contains ORF region of 234 bp. This ORF showed strong sequence similarities to the uncharacterized genes from Arabidopsis. The expression of BrTSR53 was determined by quantitative real-time PCR analysis. After 3 hr, the highest quantities of mRNA were revealed in cold and salt stress treatments. In drought stress treatments, there was the highest expression after 36 hr. Therefore, it was confirmed that the ORF in BrTSR53 should be a gene that confer increased resistance to B. rapa growing in different stress conditions. The ORF region of BrTSR53 gene was cloned into an expression vector, pYES-DEST52, and a new protein with molecular weight of 13 kDa was detected by western blot analysis. Also, stress tolerance tests showed that BrTSR53-ORF transgenic yeast exhibited increased resistance to the salt stresses compared with the control. In conclusion, the present data predicts that novel ORF in BrTSR53 can serve as an important genetic resource for abiotic stress resistance.

The next generation sequencing (NGS) has been developed rapidly in recent years, paving ways of discovering vast sequence variations among germplasms. Whole-genome sequencing was performed on the genomic DNA of Milyang23 and Gihobyeo using NGS and developed new CAPS (cleaved amplified polymorphic sequence) markers based on the single nucleotide polymorphisms (SNPs) in coding sequence between these varieties. The NGS sequencing yielded sequences of 60x coverage of the Nipponbare reference genome on average. A molecular genetic map was constructed with the recombinant inbred population derived from Milyang23/Gihobyeo cross (MGRIL) integrating the newly developed 146 CAPS makers and previously reported 219 PCR-based DNA markers. This map was applied to the detection of quantitative trait loci (QTLs) for stem internode diameters, culm length and panicle length in rice with MGRIL population. A total of 4 new QTLs were detected for stem diameter traits including the first internode diameter (I1D), second internode diameter (I2D), third internode diameter (I3D), and fourth internode diameter (I4D). Among stem diameter QTLs, qI1D5 had relatively 6.09 LOD (likelihood of odds) score and explained 8.99% of total variation. Only very small portion of SNPs through re-sequencing were used in this study. Much more markers can be developed by using SNP information acquired in this study, which will enable construction of high-density genetic map and more accurate QTL analysis of important agronomical traits with MGRIL population.