• KOREAN JOURNAL OF CROP SCIENCE
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• v.67 no.1
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• pp.41-52
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• 2022

As a cultivar of Korean wheat, 'Keumgang' wheat variety has a fast growth period and can be grown stably. Hexaploid wheat (Triticum aestivum) has moderately high salt tolerance compared to tetraploid wheat (Triticum turgidum L.). However, the molecular mechanisms related to salt tolerance of hexaploid wheat have not been elucidated yet. In this study, the candidate genes related to salt tolerance were identified by investigating the genes that are differently expressed in Keumgang variety and examining salt tolerant mutation '2020-s1340.'. A total of 85,771,537 reads were obtained after quality filtering using NextSeq 500 Illumina sequencing technology. A total of 23,634,438 reads were aligned with the NCBI Campala Lr22a pseudomolecule v5 reference genome (Triticum aestivum). A total of 282 differentially expressed genes (DEGs) were identified in the two Triticum aestivum materials. These DEGs have functions, including salt tolerance related traits such as 'wall-associated receptor kinase-like 8', 'cytochrome P450', '6-phosphofructokinase 2'. In addition, the identified DEGs were classified into three categories, including biological process, molecular function, cellular component using gene ontology analysis. These DEGs were enriched significantly for terms such as the 'copper ion transport', 'oxidation-reduction process', 'alternative oxidase activity'. These results, which were obtained using RNA-seq analysis, will improve our understanding of salt tolerance of wheat. Moreover, this study will be a useful resource for breeding wheat varieties with improved salt tolerance using molecular breeding technology.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.140-140
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• 2017

Nutritious and functional foods from crop have received great attention in recent years. Colored-grain wheat contains high phenolic compound and a large number of flavonoid. The anthocyanin and polyphenolic synthesis and accumulation is generally stimulated in response to biotic or abiotic stresses. Here, we analyzed genome wide transcripts in seedling of colored-grain wheat response to ABA and PEG treatment. About 900 and 1500 transcripts (p-value < 0.05) from ABA and PEG treatment were aligned to IWGSC1+popseq DB which is composed of over 110,000 transcripts including 100,934 coding genes. NR protein sequences of Poaceae from NCBI and protein sequence of transcription factors originated from 83 species in plant transcription factor database v3.0 were used for annotation of putative transcripts. Gene ontology analysis were conducted and KEGG mapping was performed to show expression pattern of biosynthesis genes related in flavonoid, isoflavonoid, flavons and anthocyanin biopathway. DroughtDB (http://pgsb.helmholtz-muenchen.de/droughtdb/) was used for detection of DEGs to explain that physiological and molecular drought avoidance by drought tolerance mechanisms. Drought response pathway, such as ABA signaling, water and ion channels, detoxification signaling, enzymes of osmolyte biosynthesis, phospholipid metabolism, signal transduction, and transcription factors related DEGs were selected to explain response mechanism under water deficit condition. Anthocyanin, phenol compound, and DPPH radical scavenging activity were measured and antioxidant activity enzyme assays were conducted to show biochemical adaptation under water deficit condition. Several MYB and bHLH transcription factors were up-regulated in both ABA and PEG treated condition, which means highly expressed MYB and bHLH transcription factors enhanced the expression of genes related in the biosynthesis pathways of flavonoids, such as anthocyanin and dihydroflavonols in colored wheat seedlings. Subsequently, the accumulation of total anthocyanin and phenol contents were observed in colored wheat seedlings, and antioxidant capacity was promoted by upregulation of genes involved in maintaining redox state and activation of antioxidant scavengers, such as CAT, APX, POD, and SOD in colored wheat seedlings under water deficit condition. This work may provide valuable and basic information for further investigation of the molecular responses of colored-grain wheat to water deficit stress and for further gene-based studies.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.61 no.1
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• pp.41-49
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• 2016

This study was conducted to evaluate maize downy mildew resistance using spreader row technique in Cambodia. A total of forty maize lines were used in this experiment. Seven Korean varieties and seven breeding lines showed high infection rates (80~100%) and highly susceptible (HS) to downy mildew disease in both spring and fall. Also most of nested association mapping (NAM) parent lines were highly susceptible (HS). Meanwhile three inbred lines, Ki3, Ki11, and CML228, showed highly resistant (HR) or resistant (R) in spring and moderately resistant (MR) in fall. These three lines were already known as resistant inbred lines against downy mildew disease. It appears that spreader row technique was suitable for selection of maize downy mildew resistance in Cambodia. The incidence of downy mildew was influenced by weather conditions, especially relative humidity and temperature. Among several inoculation methods to screen for downy mildew resistance, this spreader row technique is effectively and easily used in the field of Southeast Asia.

• Proceedings of the Korean Society of Crop Science Conference
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• 2004.04a
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• pp.12-27
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• 2004

Thanks to spectacular advances in the techniques for identifying proteins separated by two-dimensional electrophoresis and in methods for large-scale analysis of proteome variations, proteomics is becoming an essential methodology in various fields of plant sciences. Plant proteomics would be most useful when combined with other functional genomics tools and approaches. A combination of microarray and proteomics analysis will indicate whether gene regulation is controlled at the level of transcription or translation and protein accumulation. In this review, we described the catalogues of the rice proteome which were constructed in our program, and functional characterization of some of these proteins was discussed. Mass-spectrometry is a most prevalent technique to identify rapidly a large of proteins in proteome analysis. However, the conventional Western blotting/sequencing technique us still used in many laboratories. As a first step to efficiently construct protein data-file in proteome analysis of major cereals, we have analyzed the N-terminal sequences of 100 rice embryo proteins and 70 wheat spike proteins separated by two-dimensional electrophoresis. Edman degradation revealed the N-terminal peptide sequences of only 31 rice proteins and 47 wheat proteins, suggesting that the rest of separated protein spots are N-terminally blocked. To efficiently determine the internal sequence of blocked proteins, we have developed a modified Cleveland peptide mapping method. Using this above method, the internal sequences of all blocked rice proteins (i. e., 69 proteins) were determined. Among these 100 rice proteins, thirty were proteins for which homologous sequence in the rice genome database could be identified. However, the rest of the proteins lacked homologous proteins. This appears to be consistent with the fact that about 30% of total rice cDNA have been deposited in the database. Also, the major proteins involved in the growth and development of rice can be identified using the proteome approach. Some of these proteins, including a calcium-binding protein that fumed out to be calreticulin, gibberellin-binding protein, which is ribulose-1,5-bisphosphate carboxylase/oxygenase activate in rice, and leginsulin-binding protein in soybean have functions in the signal transduction pathway. Proteomics is well suited not only to determine interaction between pairs of proteins, but also to identify multisubunit complexes. Currently, a protein-protein interaction database for plant proteins (http://genome .c .kanazawa-u.ac.jp/Y2H)could be a very useful tool for the plant research community. Recently, we are separated proteins from grain filling and seed maturation in rice to perform ESI-Q-TOF/MS and MALDI-TOF/MS. This experiment shows a possibility to easily and rapidly identify a number of 2-DE separated proteins of rice by ESI-Q-TOF/MS and MALDI-TOF/MS. Therefore, the Information thus obtained from the plant proteome would be helpful in predicting the function of the unknown proteins and would be useful in the plant molecular breeding. Also, information from our study could provide a venue to plant breeder and molecular biologist to design their research strategies precisely.

• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.25-25
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• 2022

Drought becomes frequent and severe because of continuous global warming, leading to a significant loss of crop yield. In soybean (Glycine max [L.]), most of quantitative trait loci (QTLs) analyses for drought tolerance have conducted by investigating yield changes under water-restricted conditions at the reproductive stages. More recently, the necessity of QTL studies to use physiological indices responding to drought at the early growth stages besides the reproductive ones has arisen due to the unpredictable and prevalent occurrence of drought throughout the soybean growing season. In this study, we thus identified QTLs conferring wilting scores and moisture contents of soybean subjected to drought stress in the early vegetative stage using an recombinant inbred line (RIL) population derived from a cross between Taekwang (drought-sensitive) and SS2-2 (drought-tolerant). For the two traits, the same major QTL was located on chromosome 10, accounting for up to 11.5% of phenotypic variance explained with LOD score of 12.5. This QTL overlaps with a reported QTL for the limited transpiration trait in soybean and harbors an ortholog of the Arabidopsis ABA and drought-induced RING-D UF1117 gene. Meanwhile, one of important features of plant drought tolerance is their ability to limit transpiration rates under high vapor pressure deficiency in response to mitigate water loss. However, monitoring their transpiration rates is time-consuming and laborious. Therefore, only a few population-level studies regarding transpiration rates under the drought condition have been reported so far. Via employing an Arduino-based platform, for the reasons addressed, we are measuring and recording total pot weights of soybean plants every hour from the 1^st day after water restriction to the days when the half of the RILs exhibited permanent tissue damage in at least one trifoliate. Gradual decrease in moisture of soil in pots as time passes refers increase in the severity of drought stress. By tracking changes in the total pot weights of soybean plants, we will infer transpiration rates of the mapping parents and their RILs according to different levels of VPD and drought stress. The profile of transpiration rates from different levels of severity in the stresses facilitates a better understanding of relationship between transpiration-related features, such as limited maximum transpiration rates, to water saving performances, as well as those to other drought-responsive phenotypes. Our findings will provide primary insights on drought tolerance mechanisms in soybean and useful resources for improvement of soybean varieties tolerant to drought stress.

• Journal of The Korean Society of Grassland and Forage Science
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• v.42 no.2
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• pp.127-136
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• 2022

This study was conducted to estimate the damage of Whole Crop Maize (WCM) according to abnormal climate using machine learning and present the damage through mapping. The collected WCM data was 3,232. The climate data was collected from the Korea Meteorological Administration's meteorological data open portal. Deep Crossing is used for the machine learning model. The damage was calculated using climate data from the Automated Synoptic Observing System (95 sites) by machine learning. The damage was calculated by difference between the Dry matter yield (DMY)_normal and DMY_abnormal. The normal climate was set as the 40-year of climate data according to the year of WCM data (1978~2017). The level of abnormal climate was set as a multiple of the standard deviation applying the World Meteorological Organization(WMO) standard. The DMY_normal was ranged from 13,845~19,347 kg/ha. The damage of WCM was differed according to region and level of abnormal climate and ranged from -305 to 310, -54 to 89, and -610 to 813 kg/ha bnormal temperature, precipitation, and wind speed, respectively. The maximum damage was 310 kg/ha when the abnormal temperature was +2 level (+1.42 ℃), 89 kg/ha when the abnormal precipitation was -2 level (-0.12 mm) and 813 kg/ha when the abnormal wind speed was -2 level (-1.60 m/s). The damage calculated through the WMO method was presented as an mapping using QGIS. When calculating the damage of WCM due to abnormal climate, there was some blank area because there was no data. In order to calculate the damage of blank area, it would be possible to use the automatic weather system (AWS), which provides data from more sites than the automated synoptic observing system (ASOS).

• Journal of The Korean Society of Grassland and Forage Science
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• v.43 no.1
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• pp.11-21
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• 2023

This study was conducted to estimate the damage of Whole Crop Corn (WCC; Zea Mays L.) according to abnormal climate using machine learning as the Representative Concentration Pathway (RCP) 4.5 and present the damage through mapping. The collected WCC data was 3,232. The climate data was collected from the Korea Meteorological Administration's meteorological data open portal. The machine learning model used DeepCrossing. The damage was calculated using climate data from the automated synoptic observing system (ASOS, 95 sites) by machine learning. The calculation of damage was the difference between the dry matter yield (DMY)_normal and DMY_abnormal. The normal climate was set as the 40-year of climate data according to the year of WCC data (1978-2017). The level of abnormal climate by temperature and precipitation was set as RCP 4.5 standard. The DMY_normal ranged from 13,845-19,347 kg/ha. The damage of WCC which was differed depending on the region and level of abnormal climate where abnormal temperature and precipitation occurred. The damage of abnormal temperature in 2050 and 2100 ranged from -263 to 360 and -1,023 to 92 kg/ha, respectively. The damage of abnormal precipitation in 2050 and 2100 was ranged from -17 to 2 and -12 to 2 kg/ha, respectively. The maximum damage was 360 kg/ha that the abnormal temperature in 2050. As the average monthly temperature increases, the DMY of WCC tends to increase. The damage calculated through the RCP 4.5 standard was presented as a mapping using QGIS. Although this study applied the scenario in which greenhouse gas reduction was carried out, additional research needs to be conducted applying an RCP scenario in which greenhouse gas reduction is not performed.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.49 no.5
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• pp.423-428
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• 2004

Soybean seeds contain high amounts of isoflavones that display biological effects and isoflavone content of soybean seed can vary by year, environment, and genotype. Objective of this study was to identify quantitative trait loci that underlie isoflavone content in soybean seeds. The study involved 85 $F_2$ populations derived from Korean soybean cultivar 'Kwangkyo' and wild type soybean 'IT182305' for QTL analysis associated with isoflavone content. Isoflavone content of seeds was determined by HPLC. The genetic map of 33 linkage groups with 207 markers was constructed. The linkage map spanned 2,607.5 cM across all 33 linkage groups. The average linkage distance between pair of markers among all linkage groups was 12.6 cM in Kosambi map units. Isoflavone content in $F_2$ generations varied in a fashion that suggested a continuous, polygenic inheritance. Eleven markers (4 RAPD, 3 SSR, 4 AFLP) were significantly associated with isoflavone content. Only two markers, Satt419 and CTCGAG3 had F-tests that were significant at P<0.01 in $F_2$ generation for isoflavone content. Interval mapping using the $F_2$ data revealed only two putative QTLs for isoflavone content. The peak QTL region on linkage group 3, which was near OPAG03c, explained $14\%$ variation for isoflavone content. The peak QTL region on linkage group 5, which was located near OPN14 accounted for $35.3\%$ variation for isoflavone content. Using both Map-Maker-QTL $(LOD{\geq}2.0)$ and single-factor analysis $(P{\leq}0.05)$, one marker, CTCGAG3 in linkage group 3 was associated with QTLs for isoflavone content. This information would then be used in identification of QTLs for isoflavone content with precision

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.21-21
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• 2017

Severe lodging has recurrently occurred at strong typhoon's hitting in recent climate change. The identification of quantitative trait loci (QTLs) and their responsible genes associated with a strong culm and their pyramiding are important for developing high-yielding varieties with a superior lodging resistance. To identify QTLs for lodging resistance, the tropical japonica line, Chugoku 117 and the improved indica variety, Habataki were selected as the donor parent, as these had thick and strong culms compared with the temperate japonica varieties in Japan such as Koshihikari. By using chromosome segment substitution lines (CSSLs) in which chromosome segments from the japonica variety were replaced to them from Habataki, we identified the QTLs for strong culm on chrs. 1 and 6, which were designated as STRONG CULM1 (SCM1) and STRONG CULM2 (SCM2), respectively. By using recombinant inbred lines (BILs) derived from a cross between Chugoku 117 and Koshihikari and introgression lines, we also identified the other QTLs for strong culm on chrs. 3 and 2, which were designated as STRONG CULM3 (SCM3) and STRONG CULM4 (SCM4), respectively. Candidate region of SCM1 includes Gn1 related to grain number. SCM2 was identical to APO1, a gene related to the control of panicle branch number, and SCM3 was identical to FC1, a strigolactone signaling associated gene, by performing fine mapping and positional cloning of these genes. To evaluate the effects of SCM1~SCM4 on lodging resistance, the Koshihiakri near isogenic line (NIL) with the introgressed SCM1 or SCM2 locus of Habataki (NIL-SCM1, NIL-SCM2) and the another Koshihikari NIL with the introgeressed SCM3 or SCM4 locus of Chugoku 117 (NIL-SCM3, NIL-SCM4) were developed. Then, we developed the pyramiding lines with double or triple combinations derived from step-by-step crosses among NIL-SCM1 NIL-SCM4. Triple pyramiding lines (NIL-SCM1+2+3, ~ NIL-SCM1+3+4) showed the largest culm diameter and the highest culm strength among the combinations and increased spikelet number due to the pleiotropic effects of these genes. Pyramiding of strong culm genes resulted in much increased culm thickness, culm strength and spikelet number due to their additive effect. SCM1 mainly contributed to enhance their pyramiding effect. These results in this study suggest the importance of identifying the combinations of superior alleles of strong culm genes among natural variation and pyramiding these genes for improving high-yielding varieties with a superior lodging resistance.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.141-141
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• 2017

The ubiquitin-proteasome pathway is the major regulatory mechanism in a number of cellular processes for selective degradation of proteins and involves three steps: (1) ATP dependent activation of ubiquitin by E1 enzyme, (2) transfer of activated ubiquitin to E2 and (3) transfer of ubiquitin to the protein to be degraded by E3 complex. F-box proteins are subunit of SCF complex and involved in specificity for a target substrate to be degraded. F-box proteins regulate many important biological processes such as embryogenesis, floral development, plant growth and development, biotic and abiotic stress, hormonal responses and senescence. However, little is known about the F-box genes in wheat. The draft genome sequence of wheat (IWGSC Reference Sequence v1.0 assembly) used to analysis a genome-wide survey of the F-box gene family in wheat. The Hidden Markov Model (HMM) profiles of F-box (PF00646), F-box-like (PF12937), F-box-like 2 (PF13013), FBA (PF04300), FBA_1 (PF07734), FBA_2 (PF07735), FBA_3 (PF08268) and FBD (PF08387) domains were downloaded from Pfam database were searched against IWGSC Reference Sequence v1.0 assembly. RNA-seq paired-end libraries from different stages of wheat, such as stages of seedling, tillering, booting, day after flowering (DAF) 1, DAF 10, DAF 20, and DAF 30 were conducted and sequenced by Illumina HiSeq2000 for expression analysis of F-box protein genes. Basic analysis including Hisat, HTseq, DEseq, gene ontology analysis and KEGG mapping were conducted for differentially expressed gene analysis and their annotation mappings of DEGs from various stages. About 950 F-box domain proteins identified by Pfam were mapped to wheat reference genome sequence by blastX (e-value < 0.05). Among them, more than 140 putative F-box protein genes were selected by fold changes cut-offs of > 2, significance p-value < 0.01, and FDR<0.01. Expression profiling of selected F-box protein genes were shown by heatmap analysis, and average linkage and squared Euclidean distance of putative 144 F-box protein genes by expression patterns were calculated for clustering analysis. This work may provide valuable and basic information for further investigation of protein degradation mechanism by ubiquitin proteasome system using F-box proteins during wheat development stages.