고추역병균(Phytophthora capsici)은 고추 생육 전반에 걸쳐 병을 발생시켜 농가 소득에 큰 손실을 일으키고 있다. 고추 역병균 저항성은 양적 형질 유전자좌(Quantitative Trait Loci, QTL)에 의해 조절되며 주동 유전자는 고추의 5번 염색체에 존재한다고 보고 되었지만, 후보 유전자의 선발 및 저항성 유전자 규명 연구는 아직 초기 단계이다. 특히, 고추는 형질전환이 어려운 작물로써 병원균과의 상호작용 연구를 통한 저항성 유전자 동정에 제한이 많다. 반면 고추와 같은 가지과 작물인 담배(Nicotiana benthamiana)는 병원균 상호작용 모델로 알려져 형질전환을 통해 저항성 유전자 규명에 활용된다. 본 연구에서는 고추 역병 저항성 기작 규명을 위한 기초 연구로써, 식물 저항성 유사 유전자(Resistance Gene Analog, RGA)를 선발하고, 이들 유전자들에 대한 담배 형질전환 기법 최적화 연구를 수행하였다. 고추 5번 염색체에 존재하는 고추 역병 저항성 분자표지들을 분석하여 RGA 후보 유전자인 CaNBARC105, CaNBARC112 유전자를 동정하였다. 이들 유전자들에 대해 Agrobacterium tumefaciens를 매개체로 하여 고추 RGA가 삽입된 담배 형질전환체를 개발하였다. 형질전환 여부는 유전자 특이적인 서열을 이용한 genomic PCR과 RT-PCR 검증을 통해 이들 형질전환 된 담배들의 생육 및 발달에 영향이 없다는 것을 확인하였다. 본 연구는 향후 고추 병 저항성 후보 유전자들이 삽입된 담배 형질전환체는 고추 역병 저항성 유전자 규명 및 기작 연구에 기반이 될 것이다.

Snails and slugs are members of the mollusk phylum. They are similar in biology and structure, except slugs lack the snail's external spiral shell. Snails and slugs feed on a variety of living plants, decaying plant matter, fruit, and flowers. They are among the most bothersome creatures on horticultural crops such as Chinese cabbage, cabbage, lettuce, ginseng, lily, and orchid (Kim et al., 1990; Kim, 1992; Sorensen, 1994). Several species of

Capsicum annuum ‘Bukang’ is a resistant variety to Cucumber mosaic virus isolate-P0 (CMV-P0), CMV-P1 can overcome the CMV resistance of ‘Bukang’ due to mutations in Helicase (Hel) domain of CMV RNA1. To identify host factors involved in CMV-P1 infection, a yeast two-hybrid system derived from C. annuum ‘Bukang’ cDNA library was used. A total of 156 potential clones interacting with the CMV-P1 RNA helicase domain were isolated. These clones were confirmed by β-galactosidase filter lift assay, PCR screening and sequence analysis. Then, we narrowed the ten candidate host genes which are related to virus infection, replication or virus movement. To elucidate functions of these candidate genes, each gene was silenced by virus induced gene silencing in Nicotiana benthamiana. The silenced plants were then inoculated with green fluorescent protein (GFP) tagged CMV-P1. Virus accumulations in silenced plants were assessed by monitoring GFP fluorescence and enzyme-linked immunosorbent assay (ELISA). Among ten genes, silencing of formate dehydrogenase (FDH) or calreticulin-3 (CRT3) resulted in weak GFP signals of CMV-P1 in the inoculated or upper leaves. These results suggested that FDH and CRT3 are essential for CMV infection in plants. The importance of FDH and CRT3 in CMV-P1 accumulation was also validated by the accumulation level of CMV coat protein confirmed by ELISA. Altogether, these results demonstrate that FDH and CRT3 are required for CMV-P1 infection in plants.

Sessile organism, plants constitutively challenged with pathogens have been developed various strategies for protection, such as preformed and inducible defense mechanisms. Receptor-like Proteins(RLPs) play critical roles in defense response as well as in plant development and growth. The domain structure of RLPs consists of extracellular leucine–rich repeats, a transmembrane domain, and a short cytoplasmic tail. Here, we identified putative 170 RLP genes from pepper genome using in-house bioinformatics pipeline. The distribution of RLPs on pepper pseudomolecule showed uneven spread and a number of RLPs were physically clustered by tandem array in the specific chromosome. Motifs analysis of pepper RLPs showed conserved LRR sequences (LxxLxxLDLxxNxxxGxIP). To understand further functional and evolutionary characteristics, evolutional relationship and gene profiling analysis are on progress.

In plants, eukaryotic translation elongation factor 1B (eEF1B) is composed of three subunits, eEF1Bα, eEF1Bβ and eEF1B γ. Two subunits are nucleotide exchange subunits (eEF1Bα and eEF1Bβ) and one is a structural protein (eEF1Bγ). In the previous study, eEF1B was identified as a common host factor for several RNA viruses. To test which subunit of eEF1B is essential for Potato virus X (PVX) replication, the virus-induced gene silencing (VIGS) for eEF1Bα, β or γ was performed in Nicotiana benthamiana and green fluorescent protein (GFP)-tagged PVX was inoculated. PVX-GFP accumulation was decreased when eEF1Bβ or γ subunit was silenced, whereas eEF1Bα had no effect on PVX-GFP accumulation in inoculated leaves. Targeting induced local lesions in genome (TILLING) was performed using a Capsicum annuum EMS population to test whether mutations in eEF1Bβ subunit affect virus infection in pepper. We obtained 81 eEF1Bβ mutant lines consisted of 16,759 individuals. These mutant lines are being tested to validate the function of eEF1B β in PVX replication.

Pepper mottle virus (PepMoV) is frequently occurring virus in pepper field. PepMoV infected plants show symptoms including mosaic leaf, distortion of foliage and fruit deformation. The dominant gene Pvr7 from Capsicum annuum ‘9093’ confers resistance to PepMoV. Previous research reveals that Pvr7 is located in 10 chromosome and linked to the dominant potyvirus resistance gene Pvr4 and Tomato spotted wilt virus (TSWV) resistance gene Tsw. To identify the Pvr7 gene, we constructed an intraspecific F2 mapping population from a cross between C. annuum ‘9093’ (PepMov resistant) and C. annuum ‘Jejujaerae’ (PepMoV susceptible). Resistance of F2 plants were screened with green flouorescent protein (GFP) tagged PepMoV. Genomic DNA was extracted from F2 individuals and markers were developed using C.annuum ‘CM334’ whole genome sequence (WGS). Several single nucleotide polymorphism (SNP) markers that were co-segregated with Pvr7 were developed. We are expecting that this Pvr7 SNP marker can be used to breeding PepMoV resistant cultivars and fine mapping of Pvr7.

고추의 고유 특성인 매운맛은 태좌 조직에서 주로 발현되는 capsaicinoid라는 화학물질에 의한 것으로 알려져 있고, 이 물질을 생합성하는 유전자와 매운맛의 유무를 판별할 수 있는 분자표지까지 개발되어 있다. 그러나, 아직 매운맛 함량에 대한 유전연구와 분자적 기전은 그 연구가 깊지 못한 상황이고, 본 연구는 고추 매운맛 함량 연구를 위한 F7 RIL mapping집단을 육성하고 매운맛 함량을 비롯한 주요한 원예적 특성들을 평가하기 위해 수행되었다. 자방친인 ‘생력211’의 capsaicinoid 함량은 341.6 mg/100 g으로 ‘청양’ 고추 품종보다 높은 수준인 것으로 확인되었고, 화분친인 ‘생력213’은 무신미인 것으로 HPLC 결과와 분자표지 결과가 일치하였다. 양친을 교배 후 자가수정하여 육성된 F7 RIL 93계통들에 대해서 매운맛 함량을 분석한 결과, 최저 77.8 mg/100 g부터 최고 1046.0 mg/100 g까지로 그 변이 폭이 매우 크며, 매운 계통들 내에서의 함량 분포도는 정규곡선 양상을 보여 본 육성 집단이 향후 매운맛 함량 연구를 위해 매우 유용하게 활용될 수 있을 것으로 기대된다.

Tsw, a single dominant resistant gene against Tomato spotted wilt virus (TSWV), has been mapped on chromosome 10 in Capsicum chinense species. Previously reported molecular markers linked to the Tsw gene are not transferable for all pepper breeding materials. To develop additional markers and do genome-based fine mapping of the Tsw gene, approaches of mapping comparison, pooled transcriptome analysis, and genome walking were applied. Eleven additional SNP molecular markers tightly linked to the Tsw gene were developed using tomato and pepper whole genome sequencing databases. Among them, four SNP markers, SNP7715-1, SNP68-1, SNP17918-1, and SNP1072-1, showed no recombination in two segregating populations of F2 ‘Telmo’ (210 individuals) and ‘SP’ (843 individuals). Three scaffold sequences from the C. annuum BAC database and two BAC clones from the BAC library of C. annuum ‘CM334’ covering the Tsw gene were identified by transcriptome analysis and genome walking. A pepper scaffold sequence covering three pepper scaffold sequences was identified from a final version of the C. annuum BAC database. The Tsw gene was delimited within 149 kb by alignment analysis of two BAC clone sequences and the pepper scaffold sequence. A total of 22 predicted genes were resided in the target region between SNP7715-1 and SNP1072-1 co-segregating markers. Among them, five predicted genes showing annotations of CC/TIR-NBS-LRR resistance proteins, mRNA-6, mRNA-7, mRNA-11, mRNA-12, and mRNA-13, were identified. The transcriptome analysis and gene expression study showed that the mRNA-13 was expressed in ‘PI152225’ but was absent in ‘Special’, demonstrating the mRNA-13 could be a strong candidate gene for the Tsw gene. This result will be favorable for cloning the Tsw gene and developing cultivars which carry the TSWV-resistance gene.

본 연구는 고추의 다양한 품질 관련 특성 연구를 위한 분자육종시스템 구축의 기초 연구로서, 고추의 매운맛과 고색소, 다양한 색소, 과형 등이 다양한 고세대(F6) RIL 집단을 육성하여 그 원예적 특성을 조사하였다. 자방친으로 단고추 계통인 “만다린(Capsicum annuum L.)”을 이용하였고, 화분친으로는 “블랙클러스터(Capsicum annuum L.)”를 이용하여 F1 조합을 작성한 후, 자식으로 세대를 유지한 고세대(F6) RIL 집단 129계통들에 대해서 과실특성과 초장, 잎, 꽃 등 13가지 원예적 특성들을 평가했다. 미숙과에서 성숙과로의 색 변화의 경우 그 변화 양상이 매우 다양하여 모두 12가지 양상으로 구분해 볼 수 있었는데, 녹색에서 바로 적색으로 변화하는 양상이 51계통으로 가장 많은 분포를 차지했고, 녹색에서 진보라색으로의 변화가 45계통, 상아색에서 주황색으로 변색하는 양상이8계통 등 숙기에 따른 과색 변화가 대단히 다양한 것을 확인할 수 있었다. 숙과색은 적색과가 99계통으로 가장 많은 분포를 보였고, 주황색이 17계통, 황색이 13계통이었다. 과형은 크게 세가지로 구분되었는데, 삼각형 과실이 64계통으로 가장 많은 분포를 보였고, 타원형이 52계통, 심장형이 13계통이었다. 착과방향은 61.2%가 상향 착과성을 보였고, 38.7%가 하향이었다. 원예적 특성 평가 결과 본 F6 RIL 집단은 그 다양성이 풍부하여 향후 고추 품질 특성 연구에 유용하게 활용될 수 있을 것으로 기대된다.

Cucumber mosaic virus (CMV), which has the broadest host range among plant viruses, is a very destructive pathogen in pepper production. Various resistance sources against CMV have been identified by plant breeders. One of them is Capsicum annuum ‘Bukang’, which contains a single dominant resistance gene, Cmr1. C. annuum ‘Bukang’ is resistant to CMV-P0 strains (CMV-Kor and CMV-Fny), but susceptible to CMV-P1. CMV-P1 is a new strain recently identified in South Korea. Previously we showed that CMV-P1 RNA1 helicase domain is responsible for overcoming Cmr1 and may be play a role in viral replication and systemic infection. To identify the plant host factors involved in CMV-P1 replication and movement, we utilized a yeast two-hybrid system derived from C. annuum ‘Bukang’ cDNA library as a prey. A total of 78 potential host genes interacting with the CMV-P1 RNA1 helicase domain were isolated in the first screening, and PCR confirmation and sequencing analysis narrowed the candidates to ten genes. The candidate genes have found to be encoding acireductone dioxygenase, ADP-glucose pyrophosphorylase, ADP-ribosylation factor 1, ADP-ribosylation factor, calreticulin-3 precursor, cysteine synthase, formate dehydrogenase, histone-H3, phosphomannomutase and polyubiqutin 6PU11. Previous studies showed that these genes were involved in virus infection, replication or virus movement. To elucidate the function of these genes, VIGS and coimmumo precipitation assay is being done.

The Cmr1 gene in peppers confers resistance to Cucumber mosaic virus isolate-P0 (CMV-P0). Cmr1 restricts the systemic spread of CMV-Fny, whereas this gene cannot block the spread of CMV-P1 to the upper leaves, resulting in systemic infection. To identify the virulence determinant of CMV-P1, six reassortant viruses and six chimeric viruses derived from CMV-Fny and CMV-P1 cDNA clones were used. Our results demonstrate that the helicase domain encoded by CMV-P1 RNA1 determines susceptibility to systemic infection. To identify the key amino acids determining systemic infection with CMV-P1, we then constructed amino acid substitution mutants. Of the mutants tested, amino acid residues at positions 865, 896, 957, and 980 in the 1a protein sequence of CMV-P1 affected the systemic infection. Virus localization studies with CMV-GFP clones and in situ localization of virus RNA revealed that these four amino acid residues together form the movement determinant for CMV-P1 movement from the epidermal cell layer to mesophyll cell layers. Quantitative real-time PCR revealed that CMV-P1 and a chimeric virus with four amino acid residues of CMV-P1 accumulated more genomic RNA in inoculated leaves than did CMV-Fny, indicating that those four amino acids are also involved in virus replication. These results demonstrate that the helicase domain is responsible for systemic infection by controlling virus replication and cell-to-cell movement. Whereas four amino acids are responsible for acquiring virulence in CMV-Fny, six amino acid (positions at 865, 896, 901, 957, 980 and 993) substitutions in CMV-P1 were required for complete loss of virulence in ‘Bukang’.

Anther derived double haploids (DHs) from sweet pepper genotypes ('Special', 'Derby', 'Bossanova', 'Fiesta', 'Debora' and 'Minipaprika') were used to study the agronomic variation in 2006. Ninety-nine successful DHs regenerants (32 from 'Special', 25 from 'Derby', 23 from 'Bossanova', 10 from 'Fiesta', 6 from 'Debora' and 3 from 'Minipaprika') were transplanted at plastic house and studied on their agronomic characters. Variation in agronomic characters was observed within the DHs of each genotype. DHs obtained from 'Derby' and 'Fiesta' exhibited wide variation in fruit yield plant-1 whereas averaged fruit yield plant-1 was highest in 'Derby' (1608 g) and less variation was observed in DHs of 'Bossanova'. Based on the agronomic characters expressed in DHs population at this environment, SP55, SP56, SP60, and SP116 from 'Special', SP8, SP10, SP14, SP16, and SP34 from 'Derby', SP115, SP119, SP142, SP143, SP196, and SP199 from 'Bossanova', SP41, SP45, and SP114 from 'Fiesta', SP21 from 'Debora' and SP91 from 'Minipaprika' identified as elite inbred lines and these DH lines could be used for commercial hybrids production in sweet pepper. Genetic relationship among the selected inbred lines using molecular markers and their response to diseases are further recommended to study.

This experiment was conducted on rice (cv. 2005 Thaoi) seeds to study whether priming with deep sea water (DSW) results in enhancement of seed emergence and seedling growth and to identify the optimum concentration of Deep Sea Water (DSW) for priming. Two experiments were conducted subsequently. In experiment 1, four concentrations of the DSW (10%, 20%, 30% and 40%), and in experiment 2, five concentrations of DSW (10%, 15%, 20%, 25% and 30%) were prepared and seeds were primed for 24 hours duration at 25℃. Beside this, hydro priming with plain water was also included as a control. Experiments were laid out in Completely Randomized Design (CRD) with three replications. Result showed that 20% DSW seed priming treatment had improved the emergence, seedling height, number of roots and root length as compare to other with DSW or without DSW treatments. Beyond 20% DSW priming (i.e. 25%, 30% and 40%) were not suitable for priming the seed. On the basis of seedlings growth parameters; emergence, seedling height, root number and length, and shoot root ratio, 20% DSW priming was the best priming treatment.

This experiment was conducted on wild vegetables; Gondalbi (Cirsium setidens), Deoduck (Codonopsis lanceolata Trautv.), and Jandae (Adenophora triphylla var. Joponica Hara) seed to study whether priming with deep sea water results in enhancement of seed germination and identify the optimum concentration of the priming solution, and duration of priming. Seeds were primed with 5 various concentrations (5%, 10%, 15%, 20% and 30%) of deep sea water (DSW) in 12 hours, 24 hours, and 48 hours at 24℃. Since Jandae had seed dormancy, it was kept for four weeks in refrigerator at 2℃ after priming treatment. In Deoduck, 5 percentage DSW priming significantly improved the early germination percentage, radicle length, and plumule emergence percentage. Among the priming period of treatments, 24 hours priming showed better performance in this treatment whereas, in Jandae, 12 hours priming with 10 percentages DSW significantly improved the germination percentage and germination rate. This treatment had increased the final germination percentage by 54%, 15% and 40% compared with control, plain water and KNO3 priming respectively. But in Gondalbi, priming did not improve the germination of seed. However, among the priming treatments, 12 hours priming with 3% KNO3 and 20% DSW gave better performance. In both the wild vegetables; Deoduck and Jandae, priming in deep sea water had improved the germination percentage and germination rate as compare to plain water, KNO3, and without priming treatment. Hence the best seed priming treatment on Deoduck and Jandae are 24 hours with 5% DSW and 12 hours with 10% DSW respectively.

This experiment was conducted to study whether priming with deep sea water results in enhancement of seed germination and to identify the optimum concentration of the priming solution, and duration of priming using sweet pepper (Cv. California wonder), rice (Cv. Ilpum) and ginseng seed. Sweet pepper and rice seeds were primed with 5 various concentrations (5%, 10%, 15%, 20% and 30%) for deep sea water for 48 hours, 24 hours and 12 hours at 25℃ and ginseng seeds in 5%, 10%, 15%, 20%, 25% and 30%, and 2,4,6, and 8 electrical conductivity (EC) which were made by desalinating deep sea water. Priming in deep sea water (DSW) improved the early and final germination percentage, mean germinal on rate, emergence percentage and root and shoot length, compared with plain water, KNO3 and without priming treatments. In sweet pepper, 24 hours priming with 5 percentage DSW significantly improved the early germination percentage and radical length. It has also improved the mean germination and emergence days and early emergence percentage, compared with KNO3 and control. Whereas, in rice, 48 hours priming with 10 percent DSW significantly improved the early germination percentage, plumule emergence percentage, root length and shoot height. Hence the best seed priming treatment on sweet pepper and Rice are 24 hours with 5 percentage DSW and 48 hours with 10 percentage DSW, respectively, whereas in ginseng, priming with EC4, EC8 and 25% DSW had shown better germination.

A gene coding for the GST of cotton (Gh-5) was cloned into Escherichia coli and experssed. The enzyme remained within the cytoplasm of E. coli. An 696 bp open reading frame was in the 988 base pair fragment of the recombinant plasmid pET-30b(+). The deduced protein sequence consists of 232 amino acids and has a molecular mass of 30235.58 Da. The cloned enzyme conjugated reduced glutathione and 1-chloro-2,4-dinitrobenzene (CDNB). Plant GST cDNA was expressed in microbe and produced polypeptide had function as an enzyme.