Jae-Gyun Gwag

Genome sequence of mungbean and insights into evolution within Vigna species

Mungbean (Vigna radiata) is a fast-growing, warm-season legume crop that is primarily cultivated in developing countries of Asia. Here we construct a draft genome sequence of mungbean to facilitate genome research into the subgenus Ceratotropis, which includes several important dietary legumes in Asia, and to enable a better understanding of the evolution of leguminous species. Based on the de novo assembly of additional wild mungbean species, the divergence of what was eventually domesticated and the sampled wild mungbean species appears to have predated domestication. Moreover, the de novo assembly of a tetraploid Vigna species (V. reflexo-pilosa var. glabra) provides genomic evidence of a recent allopolyploid event. The species tree is constructed using de novo RNA-seq assemblies of 22 accessions of 18 Vigna species and protein sets of Glycine max. The present assembly of V. radiata var. radiata will facilitate genome research and accelerate molecular breeding of the subgenus Ceratotropis.

Draft genome sequence of adzuki bean, Vigna angularis

Adzuki bean (Vigna angularis var. angularis) is a dietary legume crop in East Asia. The presumed progenitor (Vigna angularis var. nipponensis) is widely found in East Asia, suggesting speciation and domestication in these temperate climate regions. Here, we report a draft genome sequence of adzuki bean. The genome assembly covers 75% of the estimated genome and was mapped to 11 pseudo-chromosomes. Gene prediction revealed 26,857 high confidence protein-coding genes evidenced by RNAseq of different tissues. Comparative gene expression analysis with V. radiata showed that the tissue specificity of orthologous genes was highly conserved. Additional re-sequencing of wild adzuki bean, V. angularis var. nipponensis, and V. nepalensis, was performed to analyze the variations between cultivated and wild adzuki bean. The determined divergence time of adzuki bean and the wild species predated archaeology-based domestication time. The present genome assembly will accelerate the genomics-assisted breeding of adzuki bean.

Development and characterization of twenty-five new polymorphic microsatellite markers in proso millet (Panicum miliaceum L.)

Millets such as proso millet have excellent nutritional properties and could become a basic resource for crop breeding programs and food diversification. In this study, 25 polymorphic microsatellite markers were developed and characterized through construction of an SSR-enriched library from genomic DNA of proso millet (Panicum miliaceum L.). In total, 110 alleles were detected, with an average of 4.4 alleles per locus. Values of major allele frequency (MAF) and expected heterozygosity (HE) ranged from 0.36 to 0.98 (mean = 0.73) and from 0.04 to 0.74 (mean = 0.37), respectively. The mean genetic similarity coefficient was 0.3711, indicating that among 50 accessions of proso millet there was wide genetic variation. The newly developed microsatellite markers should be useful tools for assessing genetic diversity, understanding population structure, and breeding of proso millet.

Assessment of genetic diversity and population structure in mungbean

This study was carried out to assess the genetic diversity and to analyze the population genetic structure for a total of 692 mungbean accessions preserved at National Agrobiodiversity Center (NAC) of the Rural Development Administration (RDA), Korea. Mungbean accessions were collected from 27 countries in nine different geographic regions, and were genotyped using 15 microsatellite markers, which were developed in our previous study. A total of 66 alleles were detected among 692 accessions at all the loci with an average of 4.4 alleles per locus. All the microsatellite loci were found to be polymorphic. The expected heterozygosity (HE) and polymorphism information content (PIC) ranged from 0.081 to 0.588 (mean = 0.345) and from 0.080 to 0.544 (mean = 0.295), respectively. Of the 66 alleles, 17 (25.8%) were common (frequency range between 0.05 and 0.5), 15 (22.7%) were abundant (frequency range > 0.5), and 34 (51.5%) were rare (frequency range < 0.05). Locus GB-VR-7 provided the highest number of rare alleles(eight), followed by GB-VR-91(six) and GB-VR-113(four). Country-wide comparative study on genetic diversity showed that accessions from the USA possessed the highest genetic diversity (PIC) followed by Nepal, Iran, and Afghanistan. And region-wide showed that accessions from Europe possessed the highest average genetic diversity, followed by accessions from the USA, South Asia, West Asia, and Oceania. Twenty-seven countries were grouped into seven clades by phylogenetic relationship analysis, but clustering pattern did not strictly follow their geographical origin because of extensive germplasm exchange between/among countries and regions. As a result of a model-based analysis (STRUCTURE) of microsatellite data, two distinct genetic groups were identified which shared more than 75% membership with one of the two genetic groups. However the genetic group pattern did not reflect their geographical origin. The Duncan’s Multiple Range Test among these two genetic groups and an admixed group, with a mean of 16 phenotypic traits, showed significant difference in 12 quantitative and qualitative traits on the basis of ANOVA. These 15 newly developed SSR markers proved to be useful as DNA markers to detect genetic variation in mungbean germplasm for reasonable management and crossbreeding purposes.

Development of a core set from a large rice collection using a modified heuristic algorithm to retain maximum diversity.

A new heuristic approach was undertaken for the establishment of a core set for the diversity research of rice. As a result, 107 entries were selected from the 10 368 characterized accessions. The core set derived using this new approach provided a good representation of the characterized accessions present in the entire collection. No significant differences for the mean, range, standard deviation and coefficient of variation of each trait were observed between the core and existing collections. We also compared the diversity of core sets established using this Heuristic Core Collection (HCC) approach with those of core sets established using the conventional clustering methods. This modified heuristic algorithm can also be used to select genotype data with allelic richness and reduced redundancy, and to facilitate management and use of large collections of plant genetic resources in a more efficient way.

Isolation and characterization of 28 polymorphic SSR loci from castor bean (Ricinus communis L.)

Castor bean (Ricinus communis) is cultivated for seed oil throughout tropical and subtropical regions but the understanding of its genetic variability is limited. Because applicable microsatellite markers are not sufficient, we isolated and characterized polymorphic simple sequence repeat (SSR) loci acquired from a microsatellite-enriched genomic DNA library of castor bean. Finally, 28 SSR loci revealed polymorphisms in a castor bean collection consisting of 72 accessions. A total of 73 alleles were detected, with an average of 3.18 alleles per locus, and the polymorphism information content (PIC) ranged from 0.03 to 0.47 (mean = 0.26). Values for observed (HO) and expected (HE) heterozygosity ranged from 0.00 to 0.19 (mean = 0.11) and from 0.04 to 0.54 (mean = 0.31), respectively. To understand genetic relationships within the castor bean collection, a dendrogram was constructed based on profiles of the 28 SSR loci. These newly developed SSRs will be useful tools for assessing genetic diversity and population structure in castor bean.

Evaluation of Genetic Diversity of Red Pepper Landraces (Capsicum annuum L.) from Bulgaria Using SSR Markers

The amount of genetic variability of a species is essential for its survival and adaptation in different environments, and studies of genetic diversity using molecular markers are necessary to understand the genetic structure of a population and to orientate effective strategies of germplasm conservation. The aim of current study was to determine the SSR markers that can be used rapidly and reliably to evaluated the pepper of Bulgaria landraces, and applied the markers to assessment of introduce genetic diversity of the pepper germplasm. We used 22 polymorphic microsatellite markers to analysis of genetic diversity within 61 pepper collection of Bulgaria landraces germplasm, all SSR primers pairs produced 82 polymorphic and reproducible amplification fragments. Results showed a relatively low level of genetic diversity among pepper accessions from Bulgaria for both molecular and morphological markers. An average value of polymorphic information contents (PIC) were 0.334 with a range of 0.061 to 0.63. The mean values of observed (HO) and gene diversity (GD) were 0.383 and 0.154, respectively, indicating a considerable amount of polymorphism within this collection. A genetic distance-based phylogeny grouped into three distinct groups according to genetic difference among the accessions. An average day of flowering time was 56.9 days with a range of 48 to 65 days. The average of fruit length and width were 6.9cm with a range 2.6 to 13.9cm, and 1.9cm with a range 0.5 to 4.2cm, respectively.

Evaluation of Resistance to Colletotrichum acutatum in Pepper Genetic Resources

Resistance of pepper (Capsicum spp.) to anthracnose (Colletotrichum acutatum) was evaluated during regeneration of Capsicum spp. in National Agrobiodiversity Center. Disease severity of 896 pepper accessions (430 accessions of C. annuum, 219 accessions of C. baccatum, 14 accessions of C. chacoense, 153 accessions of C. chinense, 70 accessions of C. frutescens, 2 accessions of C. pubescens, and unidentified 8 accessions) was investigated at 14 days after inoculation in 28 C humid chamber. Forty nine accessions of pepper germplasm were resistant to C. acutatum. Among them, nine accessions were highly resistant to C. acutatum without wounding spray inoculation. Four accessions belonged to the species C. baccatum, one accession to C. chacoense, and four accessions to C. frutescens. Forty two resistant candidate accessions were inoculated with pin-prick wounding using a syringe needle. Five accessions were resistant as a less than 3% of disease severity to C. acutatum with wounding inoculation 5 days after inoculation. All resistant accessions were C. baccatum. These five pepper germplasm might be used as breeding resources for the anthracnose resistance breeding program.

Agricultural Status in Kazakhstan Republic and Korean Strategies for Agriculture Technical Cooperation with Kazakhstan

The purpose of this paper was to introduce agricultural status of the Tajikistan republic for whom are interested in agriculture of Tajikistan. Tajikistan is situated in the south-east of Central Asia and shares borders with Afghanistan, China, Kyrgyzstan, and Uzbekistan. The total land area of the Republic is 143.1 thousands km and 93 percent of the territory is mountain, and nearly half of the country area is located at 3,000 masl and above. Agriculture accounts for approximately 21.4% of GDP in 2011, and approximately 74% of the population resides in rural areas, and 49.8% of the work force is engaged in agricultural production in 2009. Because of a little amount of precipitation, almost all of arable land(702 thousands ha, 94.6%) should be irrigated. Important crops in Tajikistan are wheat, cotton, barley, corn, potato, onion, rice, and tomato. Because of low crop productivity Tajikistan imports many agricultural products including wheat, wheat flour, and potato. The livestock industry occupied 21.7% of total agricultural product and important livestock are cattle, sheep, goat, rabbit and chicken. There are about 5,000 plant species in Tajikistan, including 650 endemic species and Tajikistan is homeland for fifty plant species including dwarf wheat, sesame, melon, radish, garlic, apple, pear, grape, and berries, etc. The total amount of conserved samples ex situ is 5317, out of which 981 are local varieties and landraces, and 1775 are breeding materials. At present, though there is no agriculture technical cooperative project between Korea and Tajikistan, it will be needed to develop cooperative projects in various field of agriculture in the near future. Plant genetic resources can be a good example of cooperative projects on the basis of mutual profit.

Evaluation of Resistance in Pepper Germplasm to Cucumber mosaic virus by High Resolution Melting Analysis

In this study, total number of 1941 Capsicum accessions conserved at RDA Genebank was evaluated for their response to Cucumber mosaic virus (CMV). These accessions were composed with 9 species originated from 89 countries, included 839 Capsicum annuum, 277 C. baccatum, 395 C. chinense, 343 C. frutescens, 49 C. pubescens, and other 38 wild pepper species (C. chacoense, C. galapagoense, etc.). Resistant to CMV was screened with the 240H02SP6 SNP marker related to the Cmr1 (Cucumber mosaic resistance 1). Eighty nine accessions of pepper germplasm were resistant to CMV based on the marker. One hundred sixty two accessions showed heterozygosity. One thousand two hundred seventy accessions were susceptible to CMV. Four hundred twenty accessions did not show distinction by 240H02SP6 marker. These 89 resistant pepper germplasm can be used in a pepper breeding program against CMV.