Jong-Min Ko

Identification and characterization of anthocyanins in yard-long beans (Vigna unguiculata ssp. sesquipedalis L.) by High-performance liquid chromatography with diode array detection and electrospray ionization/mass spectrometry (HPLC-DAD-ESI/MS) analysis.

Anthocyanins play an important role in physiological functions related to human health. The objective of this study was to investigate the profiles of anthocyanins in the immature purple pods and black seeds of yard-long beans ( Vigna unguiculata ssp. sesquipedalis L.) using high-performance liquid chromatography (HPLC) with diode array detection and electrospray ionization/mass spectrometry (DAD-ESI/MS) analysis. The individual anthocyanins were identified by comparing their mass spectrometric data and retention times. In the purple pods, five individual anthocyanins were identified: delphinidin-3-O-glucoside (2), cyanidin-3-O-sambubioside (4), cyanidin-3-O-glucoside (5), pelargonidin-3-O-glucoside (7), and peonidin-3-O-glucoside (8). From the black seed coat of the yard-long beans, seven anthocyanins were identified, including delphinidin-3-O-galactoside (1), cyanidin-3-O-galactoside (3), petunidin-3-O-glucoside (6), and malvidin-3-O-glucoside (9), together with compounds 2, 5, and 8. In this study, we report for the first time anthocyanin profiles for the pod and seed coat of yard-long beans.

Analysis of somaclonal variation through tissue culture and chromosomal localization of rDNA sites by fluorescent in situ hybridization in wild Allium tuberosum and a regenerated variant

The effects of basal media and growth regulators on callus initiation and shoot regeneration have been investigated in wild Allium tuberosum (2n = 4x = 32). Callus initiation was greatest from flower bud explants cultured on MS medium supplemented with 2,4-D and BA at 1 mg l−1 each. Maximum number of shoots was obtained from callus grown on MS medium supplemented with NAA and BA at 0.2 and 2 mg l−1, respectively. The chromosome analysis of regenerants derived from callus revealed variation in ploidy, such as 2n = 28, 29, 30, 31, 33 as well as normal tetraploid. During the culture period for two generations, one aneuploid regenerant with 2n = 30 (named At30) showed better viability and growth than tetraploid plants and other aneuploid variants. In a karyotypic analysis of At30, the chromosomal positions of 5S and 18S-5.8S-26S rDNA were physically mapped by fluorescent in situ hybridization and compared to chromosomes of wild type A. tuberosum. Both wild type A. tuberosum and At30 exhibited two sets of 5S rDNA sites, one on the proximal position of the short arm of chromosome 3, and the other on the intercalary region on the long arm of chromosome 6. There was one 18S-5.8S-26S rDNA site in the secondary constriction including flanking short chromosomal segments of satellite and terminal regions on the short arm of chromosome 8 in wild type A. tuberosum. However, At30 showed only three labelled chromosome 8 indicating that this was one of the lost chromosomes of At30.

Characterization of SMV resistance of soybean produced by genetic transformation of SMV-CP gene in RNAi

Soybean mosaic virus (SMV), a species of the Potyvirus genus in the Potyviridae family, is one of the most typical viral diseases and results in yield and quality loss of cultivated soybean. Due to the depletion of genetic resources for resistance breeding, a trial of genetic transformation to improve disease resistance has been performed by introducing the SMV-CP gene by the RNA interference (RNAi) method via Agrobacterium-mediated transformation. Among 30 transgenic plants produced, 7 lines with enough seeds were infected with SMV and two lines (3 and 4) showed viral resistance to SMV infection. In genomic Southern blot analysis, all the lines tested contained at least one T-DNA insertion. Subsequent investigation confirmed that no viral CP gene expression was detected in two SMV-resistant lines after artificial inoculation of SMV, while non-transgenic control and other transgenic lines expressed substantial amounts of the viral gene. Viral symptoms affected seed morphology, and clean seeds were harvested from the resistant lines. Also, strong viral gene expression was detected from the seeds of susceptible lines. In further generations, the same phenotypic appearance was maintained among non-transgenic and transgenic plants. Finally, the presence of helper component-proteinase (HC-Pro), known as a suppressor of gene silencing apparatus, was checked among transgenic lines. No expression of HC-Pro in resistant lines indicated that the viral CP-RNAi transformation into soybean somehow created a functional gene silencing system and resulted in a viral-resistant phenotype.

Changes in anthocyanin and isoflavone concentrations in black seed-coated soybean at different planting locations.

This study assessed the altitudinal variations in the anthocyanin and isoflavone contents of six black seed coated soybean [Glycine max (L.) Merrill] cultivars. The black soybean cultivars Heugcheong, Seonheuk, Geomjeong 1, Geomjeong 2, Cheongja 2, and Cheongja 3 were planted at Milyang (12 m above mean sea level — low altitude) and Muju (600 m — high altitude), Korea on 10 June 2005 and 2006. The total anthocyanin and isoflavone contents and individual components were investigated by reverse-phase high performance liquid chromatography (HPLC). All black soybean cultivars cultivated in high altitude possessed significantly higher total anthocyanin (p < 0.01) and isoflavone (p < 0.01) contents than those grown in low altitude. For anthocyanin composition, cyanidin-3-O-glucoside, cyanidin-3-O-galactoside, and peonidin-3-O-glucoside contents were significantly higher while delphinidin-3-O-glucoside contents was significantly lower at high altitude. The composition of individual isoflavones, 6″-O-malonyldaidzin, and 6″-O-malonylgenistin contents significantly increased at high altitude.

A New Soy-paste Soybean Cultivar, ‘Daeha 1’ with Disease Resistance, Lodging Tolerance and High Yielding

A new soybean cultivar for soy-paste, ‘Daeha 1’, was developed by soybean breeding team in the Yeongnam Agricultural Research Institute (YARI) in 2008. A promising line, SS97214-S-S-S-15, was selected from the combination between ‘Suwon192’ and a pedigree came from cross combination between ‘Jangyeobkong’ and ‘Hwaeomputkong’. It was designated as the name of ‘Milyang 164’. It had good result from regional adaptation yield trial (RYT) for three years from 2006 to 2008 and released as the name of ‘Daeha 1’. It has a determinate growth habit, white flower, grey pubescence, yellow seed coat, yellow hilum, large spherical seed (25.4 grams per 100 seeds). ‘Daeha 1’ is tolerant to soybean mosaic virus and bacterial pustule, the major soybean disease in Korea. The average yield of ‘Daeha 1’ was 2.62 ton per hectare in the regional yield trial (RYT) carried out for three years from 2006 to 2008, which was 5 percent higher than that of check cultivar, ‘Taekwangkong’.

Evaluation of soybean cultivars for resistance to Phomopsis seed decay in Korea

Phomopsis seed decay (PSD), primarily caused by Phomopsis longicolla, is one of the most important seed-borne diseases and causes serious seed yield loss in soybean. This study was performed to evaluate reactions to P. longicolla in Korean soybean major elite cultivars, which were mainly used for parents of genetic mapping populations. The natural incidence of P. longicolla and other seed-borne fungi was determined in the fields at three different locations in South Korea during 2009–2010. The significant differences in sensitivity to seed-borne diseases were shown among cultivars. Taekwangkong exhibited the greatest resistance to P. longicolla with average incidence of 0.33% and other seed-borne fungi with average incidence of 6.17%. Moreover, Taekwangkong was free of P. longicolla infection both in Milyang and in Daegu. To confirm the effective resistance source, the Korean virulent strain of P. longicolla, SSLP-3, was inoculated artificially on soybean of R4–R7 growth stage in the greenhouse. Taekwangkong exhibited a higher level of resistance to P. longicolla with significantly lower incidence (8.67%) than any other Korean elite cultivars (78.0–99.33%) and the previously reported resistant PI genotypes (35.0–55.67%). Further verification of resistance in Taekwangkong to P. longicolla by testing germination vigor of healthy seeds in vitro showed a higher germination rate than those of the susceptible cultivars. It could be suggested that Taekwangkong is a newly identified resistance source and the better source of resistance to P. longicolla to develop breeding populations for exploiting resistance gene(s) in further studies.

A New Soybean Variety, ‘Joongmo 3009’ with Green Cotyledon, Black Seed Coat, Disease Tolerant, and High Yield

A new soybean variety, ‘Joongmo 3009’ (Milyang 222) was developed at the National Institute of Crop Science (NICS) in 2012. ‘Joongmo 3009’ was released by pedigree selection from the cross between ‘Cheongja 2(Milyang 121)’ and ‘Daemangkong’. It has determinate growth habit, white flower, brown pubescence, brown pod color, green seed coat, green cotyledon, spherical seed shape, oval leaf shape and large seed size (29.3 grams per 100 seeds). It was late 16 days in maturing date than the check cultivar ‘Cheongjakong’. The average yield of ‘Joongmo 3009’ was 2.91 ton per hectare, which was higher 36 percentage than the check variety, in the regional yield trials carried out in three adaptable locations of Korea from 2010 to 2012. The number of breeder’s right is ‘5474’

Cloning and Characterization of Soybean IFS (Isoflavone Synthase) Genes from Korean Cultivar, Sinpaldalkong

Two genes, SinIFS1 and SinIFS2 from Korean soybean cultivar, Sinpaldalkong known as one of isoflavonerich cultivars, were cloned with PCR and degenerate primers. The sequences of two genes were analyzed with previously reported IFS genes of leguminous plants and their expression pattern in various environmental conditions was surveyed. The genomic clone of SinIFS1 contained 1,828bp nucleotides and encoded a polypeptide of 521 amino acids, and 1912bp nucleotides and a polypeptide of 521 amino acids for SinIFS2. Both genes included several conserved motifs, oxygen binding and activation (A/G-G-X-E/D-T-T/S), ERR triad (E...R....R), and heme binding (F-X-X-G-X-R-X-C-X-G) domain, which are typical in any member of cytochrome P45O superfamily. Very high sequence homology (>98%) was observed in the comparison with other IFSs of legumes. In the northern blot analysis to check the expression and increase of SinIFS1 to various environmental renditions (low temperature, light, dark, UV, and fungal elicitor), the most significant induction, more than 6 times of transcript level compared to the dark treatment as a control, was observed from the fungal elicitor treatment. The next up-regulated expression was from UV treatment (4), low temperature and light conditions.