Hyun Ran Kim

Production System of Virus-free Apple Plants Using Heat Treatment and Shoot Tip Culture

Gunsup Lee*, Jeong Hee Kim*, Hyun Ran Kim, Il Sheob Shin, Kang Hee Cho, Se Hee Kim, Juhee Shin and Dae Hyun Kim** Fruit Research Division, National Institute of Horticultural and Herbal Science, Rural Development Administration, Suwon 440-706, Korea Apple Research Station, National Institute of Horticultural and Herbal Science, Rural Development Administration, Gunwi 716-812, Korea Future & Creation Strategy Team, Rural Development Administration, Suwon 441-707, Korea (Received on November 12, 2013; Revised on November 21, 2013; Accepted on November 25, 2013)

Characterization of Prunus necrotic ringspot virus Isolate from Peach in Korea

In this paper, we report a characterization of Prunus necrotic ringspot virus (PNRSV) isolate. The virus was identified from `Yumyeong` peach showing mild mosaic on leaves in commercial orchard of `Umsung`, Chungbuk province in Korea. The virus isolate produced ringspot symptom on the inoculated cotyledons and systemic mosaic and malformation on the upper leaves of Cucumis sativus. Systemic mottles were appeared in Chenopodium quinoa. When the buds of the virus infected stem were grafted on the healthy young Prunus persica GF305 seedlings, line pattern with mosaic appeared within 3 months. Isometric virus-like particles were found in parenchyma cells and plasmodesmata of C. sativus leaves inoculated mechanically with the virus. The cDNA fragments of PNRSV coat protein (CP) region, approximately 675bp, were synthesized from genomic RNA extracted from virus-infected leaves by RT-PCR using specific primer pairs. Partial nucleotide sequences of the CP regions were determined and analyzed with the known PNRSV. The CP gene of PNRSVKorea isolates showed 93.9~94.7% similarity to the 4 known PNRSV isolates.

Comparison of transcriptome analysis between red flash peach cultivar and white flash peach cultivar using next generation sequencing

Differences of gene expression between red flash peach cultivar and white flash peach cultivar were investigated by Nest-generation sequencing (NGS). EST from the red flash peach cultivar and white flash peach cultivar were selected for nucleotide sequence determination and homology searches. The levels of transcripts coding for proteins involved in pathogenesis related proteins, temperature stress, ethylene signal pathway were significantly higher in white flash peach cultivar than in red flash peach cultivar. On the other hand, the up-regulation of proteins involved in anthocyanin and flavonol biosynthesis and protein degradation and sorbitol metabolism were observed in red flash peach cultivar. Chalcone synthase was preferentially expressed in the red flesh peach cultivar, agreeing with the accumulation of anthocyanin and expression of other previously identified genes for anthocyanin biosynthesis. Anthocyanin pathway related genes CHS, F3H, DFR, LDOX, UFGT differentially expressed between red flash peach cultivar and white flash peach cultivar. These results suggest that red flash peach cultivar and white flash peach cultivar have different anthocyanin biosynthesis regulatory mechanisms.

Identification of genes induced by Venturia nashicola in indigenous Korean pear ‘Hwangsilri’

Indigenous Korean pear ‘Hwangsilri’ leaves were inoculated with Venturia nashicola to build expressed sequence tags (ESTs) database as resources for transcripts induced in the inoculated leaves. After performing subtractive suppression hybridization using cDNA of the inoculated and uninoculated leaves harvested at 1, 48, and 96 hours after inoculation, 159 (1 hour), 384 (48 hours), and 110 clones (96 hours) were selected and sequenced. BLASTX searches each cDNA library against Genbank revealed 15, 50, and 22 unique sequences at 1, 48, and 96 hours after inoculation, respectively. Most highly represented ESTs at 1 hour after inoculation were photosynthesis- and senescence-related genes such as light harvesting chlorophyll a/b binding protein, ribulose-bishosphate carboxylase/oxyganase, and senescence-associated protein. Cytochrome P450-like TATA box binding proteins and manganese superoxide dismutase associated with the defense response, biotic and abiotic stresses were differentially expressed at 1 hour after inoculation. Although more than 50% of ESTs at 1 and 48 hours after inoculation were also associated with photosynthesis- and carbon fixation-related genes, defense-related genes were annotated as 14 clones at 48 hours after inoculation. Six ESTs associated with pathogen-defense and biotic and abiotic stresses were expressed only at 48 hours after inoculation. At 96 hours after inoculation, seven ESTs were involved in defense-response and biotic and abiotic stresses, and three out of the seven ESTs such as cyclophilin, F-box family, and leucine-rich repeat receptor-like kinase (LRR-RLK) were uniquely expressed. Quantitative real-time polymerase chain reaction analysis revealed that the transcripts for aldo-keto reductase (AKR) and LRR-RLK were highly expressed in the incompatible interaction. AKR gene was more highly expressed in resistant cultivar ‘Hwangsilri’ and moderately susceptible one ‘Gamcheonbae’ than in other cultivars. LRR-RLK showed differentially higher expression pattern in inoculated than in uninoculated leaves of susceptible cultivars including ‘Gamcheonbae’ and ‘Hwangkeumbae’. The ARK and LRRRLK genes were found to be associated with plant defense mechanism. However, more detailed further study using transformants introgressed these genes is required to understand how these genes are expressed and regulated by infection with V. nashicola.

Plant regeneration and transformation of grape (Vitis labrusca L.) via direct regeneration method

Abstract Efficient regeneration methods and transformation system are a priority for successful application of genetic engineering to vegetative propagated plants such as grape ( Vitis labrusca L.). This research is to establish shoot regeneration system from plant explants for ‘Campbell Early’, ‘Tamnara’, ‘Heukgoosul’, ‘Heukbosek’ using two types of plant growth regulators supplemented to medium. The highest adventitious shoot regeneration rate of 5% was achieved on a medium containing of Murashige and Skoog (MS) inorganic salts and Linsmaier and Skoog (LS) vitamins, 2 mg/L of TDZ and 0.1 mg/L of IBA. Leaf tissue of ‘Campbell Early’, was co-cultivated with Agrobacterium strains, LBA4404 containing the vector pBI121 carrying with CaMV 35S promoter, gus gene as reporter gene and resistance to kanamycin as selective agent, the other Agrobacterium strains, GV3101 containing the vector pB7 WG2D carrying with mPAP1-D gene. mPAP1-D is a regulatory genes of the anthocyanin biosynthetic pathway. ‘Campbell Early’ harboring