When the rice blast fungus attacks rice, fungal proteins are secreted into the plant apoplast to facilitate infection. The rice plant recognizes such secreted proteins, which result in the induction of defense responses. However, the molecular mechanisms of how rice plant recognizes secreted proteins remain elusive. Here, we report that a small, secreted protein, Magnaporthe oryzae snodprot1 homolog (MSP1), is recognized by rice plants and triggers host cell death and defense responses. Furthermore, pre-treatment of rice with Domain II, elicitor-active epitope of MSP1, induces resistance to the pathogen KJ301. We demonstrated that secretion of MSP1 into the apoplast is prerequisite for triggering cell death and activating defense-related gene expression, suggesting that it is recognized by a receptor in the host plasma membrane. Through comprehensively analysis of transcriptional profile in rice leaves and suspension cultured cells (SCCs) in response to exogenous MSP1 and Domain II treatment using 60K Agilent microarray chip, we found that 27 signaling genes, such as F-box(6), MAPK(4), protein kinase(11), transcription factor(6), were up-regulated in leaves and SCCs and six protein kinases were targeted into plasma membrane. Thus, we suggest that some of these genes may act as receptor of MSP1 in response to exogenous MSP1 treatment. Expression pattern of candidate genes was further checked in response to different environment cues using open rice data. These results demonstrate that these genes may be also involved in the signaling in response to cold stress, root-JA treatment and brown plant hopper (BPH) attack.

Nitrogen in rice paddy soils and utilized as the major source for N-assimilation in rice crops. In roots, transcriptional activities of ammonium uptake and assimilation genes are highly sensitive to the availability of exogenous ammonium. However, little is known about the transcription factor genes that regulated by ammonium supply and its role to roots and plant developments. To study the transcription factor genes that involved in Ammonium response, two weeks old rice seedlings treated using Ammonium from 0 to 3 hours. Total RNA collected from each sample and samples were prepared for Agilent 8x60K microarray system. Based on the microarray data, we select transcription factor genes that highly affected by ammonium and selected knock out mutant candidates that used for phenotype screening.

The wild relatives of soybean [Glycine soja Sieb. and Zucc.] have curly/wavy nature whereas cultivated varieties are upright. Such morphological characteristics have agronomic importance too. To investigate the molecular mechanism of development contributing to coiled morphology, screening was carried out to look for Arabidopsis mutants in activation tagging lines obtained by activation T-DNA treatment that have curly/wavy morphology. A mutant named Coiled Branch 1 (cbr1), is found to have a wavy and curly morphology with coiling branches. Plasmid rescue and genomic southern blot analysis revealed the site of T-DNA insertion in the genome. RT-PCR was performed to monitor expression levels of the genes adjacent to the T-DNA integration sites, and showed the activation of an E3 ubiquitin ligase gene. Database search showed that the gene with the RING domain belongs to a family of E3 ubiquitin ligases. Complementation test by overexpression and RNA interference of the gene was also carried out. The complementation test results showed that the novel gene activation tagging affected the cbr1 mutant phenotypes. Ubiquitylation has been linked virtually to every cellular process including plant development. E3 ubiquitin ligase has been reported to recognize target proteins that are to be ubiquinated for further degradation by the proteasome complex. Further, more detailed studies are needed to identify the specific substrate(s) of the novel E3 ubiquitin ligase gene.

Drought and high salinity are the most important abiotic factors limiting plant development, growth, and crop productivity in agriculture (Munns and Tester 2008, Sengupta and Majumder 2009, Zhu 2002). As sessile organisms, plants are frequently exposed to drought and high salinity conditions, which alter water potential and cause osmotic stress, leading to serious damage to plant tissues (Bartels and Sunkar 2005, Boudsocq and Lauriere 2005). During exposure to water stress, plants display many physiological changes, such as reduction of water content, closure of stomata, and decreased cell enlargement and growth. In addition, severe and continuous water stress in plants causes the cessation of photosynthesis and disturbance of metabolism, and finally results in death (Nath et al. 2005, Shao et al. 2008). To adapt to these abiotic stress conditions, plants show a variety of responses, including the accumulation of abscisic acid (ABA) and expression of a large number of stress-related proteins (Krasensky and Jonak 2012, Lee and Luan 2012, Skriver and Mundy 1990, Stewart and Lee 1974). Although the cellular and molecular responses to environmental stress are well studied (Hasegawa et al. 2000, Thomashow 1999), the mechanisms underlying the functional modifications caused by osmotic stress are yet to be clarified, because of the complexity at the cellular level as well as at the whole plant level (Ashraf and Harris 2004, Flowers 2004, Foolad et al. 2003a, 2003b, Xiong et al. 2002).

The circadian clock control of CONSTANS (CO) transcription and the light regulation of CO stability coordinately regulate photoperiodic flowering by triggering rhythmic expression of the floral integrator FLOWERING LOCUS T (FT). The diurnal pattern of CO accumulation is modulated sequentially by distinct E3 ubiquitin ligases, such as HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENES 1 (HOS1) in the morning, FLAVIN-BINDING, KELCH REPEAT, F-BOX 1 (FKF1) in late afternoon, and CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1) at night. In particular, CO is stabilized by FKF1 in late afternoon only under long days. Here, we show that CO abundance is not simply regulated by the E3 enzymes in a passive manner but also self-regulated actively through dynamic interactions between two CO isoforms. CO alternative splicing produces two protein variants, the full-size COa and the C-terminally truncated COb. Notably, COb, which is resistant to the E3 enzymes, induces the interactions of COa with CO-destabilizing HOS1 and COP1 but inhibits the association of COa with CO-stabilizing FKF1. These observations demonstrate that CO plays an active role in sustaining its diurnal accumulation dynamics in Arabidopsis photoperiodic flowering.

Several E3 ubiquitin ligases have been associated with the response to abiotic and biotic stresses in higher plants. Here, we report that the hot pepper (Capsicumannuum) abscisic acid (ABA)-InsensitiveRINGprotein1gene(CaAIR1) is essential for a hypersensitive response to drought stress. CaAIR1 contains a C3HC4-type RING finger motif, which plays a role for attachment of ubiquitins to the target protein, and a putative transmembrane domain. The expression levels of CaAIR1 are upregulated in pepper leaves by ABA treatments, drought, and NaCl, suggesting its role in the response to abiotic stress. Our analysis showed that CaAIR1 displays self-ubiquitination and localized in the nucleus. We generated CaAIR1-silenced peppers via virus-induced gene silencing (VIGS) and CaAIR1-overexpressing (OX) transgenic Arabidopsis plants to evaluate their responses to ABA and drought. VIGS of CaAIR1 in pepper plants conferred an enhanced tolerance to drought stress, which was accompanied by low levels of transpirational water loss in the drought-treated leaves. CaAIR1-OX plants displayed an impaired sensitivity to ABA during seed germination, seedling, and adult stages. Moreover, these plants showed enhanced sensitivity to drought stress because of reduced stomatal closure and decreased expression of stress-responsive genes. Thus, our data indicate that CaAIR1 is a negative regulator of the ABA-mediated drought-stress tolerance mechanism.

Arsenic (As) is accumulated in rice grain due to environmental reasons such as polluted ground water and soil, and As toxicity constitutes a serious threat to human health. However, the accurate information required for understanding As-responsive mechanisms remain mostly unknown in rice. Here, we performed the comparative genome-wide transcriptome analysis between As tolerance type (ATT) rice mutant induced by γ-irradiation and its wild type (WT). As compared to WT after As treatment of 150 ppm, ATT exhibited the phenotypic differences such as vigorous growth in shoots and root hairs, and low accumulation of H2O2 in rice roots. In transcriptome analysis, we found between WT and ATT that As toxicity commonly affected to inhibit gene regulations involved in photosynthesis, mitochondrial electron transport and lipid biosynthesis metabolism. While, many genes associated with cysteine synthesis metabolism considerably up regulated in both As-treated plants. Additionally, we found the potential As tolerance-related genes involved in abiotic stress-responsive mechanism and RNA-protein synthesis for protein degradation and modification. To further analyzes the genetic variations of As-responsive genes, the DNA polymorphic DEGs associated with oxidoreductase significantly distributed in ATT more than in WT.

Enhancing yield has been a major challenge of agriculture. In rice, tiller number is one of the important biomass and yield components. A maize mutant grassy tillers1 (gt1) increases lateral branches in maize. The GT1 gene encodes a class I homeodomain leucine zipper (HD-Zip) protein. In maize, the gt1 expression is induced by shading and is dependent on the activity of teosinte branched1 (tb1), a major domestication locus controlling tillering and lateral branching. To estimate the biological role and agricultural utility of gt1 in rice, rice homologue (OsGT1) has been isolated and its overexpressors and RNAi lines were generated. Field data showed that OsGT1 overexpressors reduced tillers and panicles while RNAi lines increased them, compared to wild type. Shade signal is an important factor in determining lateral branching. To understand the relationship between OsGT1 and shade avoidance, plants have been grown under 50% shading in the field. Also, double genetic combinations with phytochrome mutants (phyA, B, and C) are being examining for tillering phenotype. These ongoing researches will provide insights in determining the action of OsGT1 on branching and shade avoidance in rice.

Lesion mimic mutants commonly display spontaneous cell death in pre-senescent green leaves under normal conditions, without pathogen attack. Despite molecular and phenotypic characterization of several lesion mimic mutants, the mechanisms of the spontaneous formation of cell death lesions remain largely unknown. Here, we examined the rice lesion mimic mutant spotted leaf3 (spl3). In mutants grown under a light/dark cycle, spl3 mutants appeared similar to wild type at early developmental stages, but lesions gradually appeared in the mature leaves close to heading stage. By contrast, in mutants grown under continuous light, severe cell death lesions formed in developing leaves, even at the seedling stage. Histochemical analysis showed that hydrogen peroxide accumulated in the mutants, likely causing the cell death phenotype. By map-based cloning and complementation, we showed that a 1-bp deletion in the first exon of Oryza sativa Mitogen-Activated Protein Kinase Kinase Kinase1 (OsMAPKKK1)/OsEDR1/ OsACDR1 causes the spl3 mutant phenotype. We found that the spl3 mutants were insensitive to abscisic acid (ABA), showing normal root growth in ABA-containing media and delayed leaf yellowing during dark-induced and natural senescence. Expression of ABA signaling-associated genes was also less responsive to ABA treatment in the mutants. Furthermore, the spl3 mutants had lower transcript levels and activities of catalases, which scavenge hydrogen peroxide, probably due to impairment of ABA-responsive signaling. Finally we discuss a possible molecular mechanism of lesion formation in the mature leaves of spl3 mutants.

Common wheat has complex genome composition of homoeologous hexaploid (AABBDD, 2n = 6x = 42) and each homoeologous genome has high similarity. Due to these complexity, wheat genome study is a large challenge to researchers for genomic and genetic study. We analyzed expressions of individual wheat genome and rye genome specific transcripts using custom array with 2BS.2RL wheat-rye translcoation. Genomic probes were synthesized within each diploid progenitors (AA, BB, DD, 2n = 14, respectively) of wheat, common wheat, and rye (RR, 2n = 14). Total RNA isolated from seedlings of T. urartu, Ae. speltoides, Ae. squarrosa, ‘Chinese Spring’, ‘Chaupon’, and 2BS.2RL were hybridized on arrays. Each homoeologous gene differentially expressed in hexaploid wheat and rye were identified on the custom array and the transcripts were clustered based on hybridization values. qRT-PCR was performed to verify the custom array result with a set of five genes by highly replicated experiments (three biological and three technical replications). The qRT-PCR results demonstrated genome specific expression of five genes in sympathy with array results. Here we provide information of each individual genome specific transcripts and it will we a useful data to study complex wheat genome compositions.

Nitrogen is a key component in the growth of crop plant. To increase the yield of crops, an enormous amount of nitrogen fertilizer is currently being used, which increases the total production cost and leads to environmental pollution by the residual nitrogen sources. For these reasons, researchers have tried to improve the crop’s nitrogen use efficiency (NUE) as a solution for reducing the amount of nitrogen fertilizer used. MicroRNAs are a class of small non-coding RNAs regulating the expression of target genes. Recent studies suggested that the expression pool of microRNAs changes in response to a variety of nutrient deficiencies and that such changes play important roles in adapting to or resisting the consequential nutritional stresses. Here, we aim to identify and characterize rice microRNAs whose expression changes upon nitrogen starvation and re-supplementation. By applying RNA-Seq, we observed that the expression of a set of genes involved in nitrogen assimilation was altered in response to nitrogen deprivation. We also found that a considerable number of microRNAs exhibited dynamic expression changes in a nitrogen supply state-dependent manner and that the expression of genes targeted by those differentially regulated microRNAs was altered reciprocally. Our study suggests that microRNAs may have roles in regulating the response of rice to nitrogen supply state and subsequently modulating NUE.

Post-translational modifications of nucleosomal core histones play important roles in biological processes via altering chromatin structure and creating target sites for proteins acting on chromatin. Molecular genetic studies with Arabidopsis have verified several epigenetic factors that regulate flowering time. However, the roles of chromatin remodeling factors have not been well explored in rice. Here, we identified chromatin remodeling factors, OsVIL1, 2, and 4 (Oryza sativa VIN3-LIKE) genes, that regulate grain yield. OsVIL proteins contain a plant homeodomain (PHD) finger, which is a conserved motif of histone binding proteins. We showed that plant height and number of spikelets per panicle were increased in the OsVIL2-overexpression (OsVIL2-OX) and osvil4 plants, respectively. Each mutants (OsVIL2-OX and osvil4) exhibited longer internodes and thicker stems than wild type controls. Histochemical analysis revealed that cells are smaller in OsVIL2-OX and osvil4 plants. We performed an RNA-seq using 1st internodes of WT and OsVIL2-OX stems and got the suppressed target genes in the OsVIL2-OX. OsCKX2, which encodes cytokinin oxidase/dehydrogenase is one of the suppressed genes in the OX plants and we verified decrease of that gene using qRT-PCR and closed chromatins of OsCKX2 were enriched in the OX plants by using ChIP. As results of these, cytokinins were enriched in the OX plants. These demonstrate that OsVIL2 and OsVIL4 antagonistically regulate plant height and number of spikelets by controlling cytokinin contents. Like OsVIL2-OX and osvil4 plants, besides, OsVIL1-OX plants were also shown increased plant height and biomass. We propose that OsVILs may be used for improving grain yield by increasing biomass.

본 과제는 “고추 육종가 맞춤식 고효율 분자육종시스템 실용화” 과제의 주관과제인, 고추 탄저병 및 CMV 저항성 마커 개발과 복합내병성 품종 육성(고추와 육종, 윤재복)으로, 세부과제인 고추 유용 분자표지의 foreground selection용 multiplexing 기술 개발(전북대학교, 이준대)과 협업을 통해 2015~2017 년까지 탄저병과 오이모자이크바이러스(cucumber mosaic virus, CMV)에 대한 복합내병성 품종 개발을 목표로 하고 있다. 탄저병과 CMV는 국내외에서 심각한 문제를 일으키고 있는 병원체로, 저항성 품종 육성 효율을 높이기 위해서는, 탄저병 저항성 연관 신규 분자표지와 CMV 강병원성 계통(기존 저항성 Cmr1 극복 CMV, CMV-P1)에 대한 저항성 연관 분자표지 개발이 필요하다. 본 과제의 성공적인 수행을 위해 현재까지 진행된 연구결과는 다음과 같다. 탄저병 저항성 연관 신규 분자표지 개발의 경우, 탄저병 및 CMV복합 CMS모계(B)와 부계(C) 계통, GMS 모계는 각각 BC1F3와 F4, F5, BC1F6 까지 세대 진전하였다. 탄저병과 바이러스에 단독 혹은 복합 내병성을 지닌 CMS와 GMS 모계, 탄저병저항성 C계통 간에 156개 교배조합을 작성하였고, 시교 사업은 경북, 경남, 충북, 충남, 전남, 전북, 강원, 인천, 제주지역을 포함하는 138개 지역에 수행하고 있다. CMV-P1 저항성 연관 분자표지 개발의 경우, 국내에서 분리된 18개 CMV 분리의 저항성 정도를 평가하여 4가지 유형으로 분류하였고, 이들을 이용해 고추유전자원의 CMV 저항성을 조사한 다음 CMV 병원형 판별 품종 후보를 선발하였다. 한편, 고추 포장에서 CMV에 강 저항을 보인 개체의 후대를 대상으로 CMV-P1대한 저항성 유전 분석을 수행하였고, 분자표지 검정을 통해 저항성과 연관된 후보 마커를 선발하였다. 금년 하반기에는 새로운 탄저병 저항성 마커 개발을 위한 저항성 유전분석 및 분리집단을 선발할 예정이며, 차년도에는 탄저병 및 CMV복합 계통의 세대진전과 신규교배 조합을 작성할 예정이다. 또한 새로운 탄저병 저항성 분자표지 개발 및 CMV-P1 저항성 연관 후보 분자표지를 이용해 CMV 병원형 판별 계통을 최종적으로 선발하고자 한다.

Nitrogen is an essential nutrient in plants including many crops. The storage and remobilization of nitrogen constitutes the main metabolic process for growth and development of plants. Ureide pathway is the lately characterized metabolic route for purine degradation and is conserved in plants, as well as some bacteria and fungi. The catabolic pathway catalyzes in a stepwise manner a conversion of N-rich uric acid into glyoxylate, with the release of ammonia, and plays a pivotal role in the storage and recovery of nitrogen from metabolites. In Next Generation BioGreen21 project, we aim to understand structural and functional features of enzymes involved in this nitrogen recycling pathway, by using genes from Arabidopsis thaliana. In this study, we report our current progress on this project including two different enzymes; ureidoglycine aminohydrolase (UGlyAH), and ureidoglycolate amidohydrolase (UAH). In UGlyAH, the metal-binding site plays a crucial role in catalysis, with a release of ammonia. We were able to characterize catalytic residues in the active site and provides a detailed view of a metal-dependent enzyme mechanism. Recently, we were able to characterize structural properties of UAH. Based on our analysis, we are performing enzymatic analysis to identify functional aspects of the enzyme. Taken together, these studies would provide a novel functional feature of the enzymes involved in the nitrogen recycling pathway and could serve as a framework to develop crops with an enhanced N-efficiency.

차세대 DNA 염기서열 분석장비 (NGS)의 발달은 유전체 대상의 DNA 정보 생산에 필요한 가격과 시간을 획기적으로 단축시켰고, 그 결과로 많은 식물들의 신규 유전체 정보가 생산 되고 있다. 또한 transcriptome, non-coding RNAs, methylome 등의 NGS기반의 데이터들은 유전체 sequence내에 유전자의 위치탐색과 유전자간 또는 유전자와 regulatory element 간의 관계를 규명하여 유전체에 대한 통합적 이해를 돕고 있다. 벼, 콩, 옥수수, 토마토, 고추, 배추 등 주요 농업작물 표준유전체 정보의 완성은 유전체 정보를 분자육종 이용할 수 있는 기반을 제공하였으며, NGS기술 (resequencing 또는 Genotype-by-Sequencing)을 통한 다양한 유전자원대상의 유전변이 정보의 생산은 유전체 정보를 육종에 적극 활용하여 중요 농업 형질과 연관된 유전적 변이를 발견하고 이를 작물개량에 활용할 수 있는 환경을 제공하고 있다. 유전체기반 분자육종시스템은 분자육종의 현장에서 효율적이고, 실용적으로 사용될 수 있는 시스템을 개발하기 위해 3가지의 목표를 가지고 수행한다. 1) 각기 산재되어있는 다양한 유전체정보 (유전체, 전사체, SNP정보, 분자마커 정보, 표현형 정보 등)를 수집하여 통합 유전체 데이터베이스화 하여 시스템 내에서 유전체, 전사체 정보를 정보를 비교, 분석이 가능한 형태로 운영하며 상호 연결된 정보를 제공하도록 구축한다. 2) 또한 최근 들어 농업에 적극 활용되는 NGS기반의 SNP genotyping에 필요한 효율적 파이프라인을 제공하여, GBS 또는 resequencing 기반의 데이터를 효율적으로 분석하고 그 결과를 토대로 genetic map구축, QTL동정, association mapping, 분자마커 개발 등에 효율성을 주는 시스템을 개발하고 3) 유전체정보와 변이정보를 연동하여 visualization 할 수 있는 브라우저와 분자마커 개발에 필요한 도구의 개발이다. 통합유전체 데이터베이스, 효율적 genotyping 시스템, 통합브라우저 등의 구축은 데이터의 생산과 분석에 표준화된 지표, 용이성을 제공하여 고도화된 유전체 정보를 분자마커 개발, QTL 탐지, 후보 유전자 동정 등 분자육종에 효율적으로 활용할 수 있게 하며, 이를 통해서 분자육종의 선진화와 종자산업의 활성화에 기여하고자 한다.

Seed germination is a key developmental transition that initiates the plant life cycle. The timing of germination is determined by coordinated action of two phytohormones, gibberellin (GA) and abscisic acid (ABA). In particular, ABA plays a key role in integrating environmental information and inhibiting the germination process. Utilization of embryonic lipid reserves contributes to seed germination by acting as an energy source, and ABA suppresses lipid degradation to modulate the germination process. Here, we report that the ABA-responsive R2R3-type MYB transcription factor MYB96, which is highly expressed in embryo, regulates seed germination by controlling the expression of ABA-INSENSITIVE 4 (ABI4). In the presence of ABA, germination was accelerated in MYB96-deficient myb96-1 seeds, whereas the process was significantly delayed in MYB96-overexpressing activation-tagging myb96-ox seeds. Consistently, myb96-1 seeds degraded a larger extent of lipid reserves even in the presence of ABA, while reduced lipid mobilization was observed in myb96-ox seeds. MYB96 directly regulates ABI4, which acts as a repressor of lipid breakdown, to define its spatial and temporal expression. Genetic analysis further demonstrated that ABI4 is epistatic to MYB96 in the control of seed germination. Taken together, the MYB96-ABI4 module regulates lipid mobilization specifically in the embryo to ensure proper seed germination under suboptimal conditions.

유전자 가위(Engineered nuclease)는 최근 유전자의 특정염기서열을 인식하여 목적 유전자 부위만을 정확히 편집하여 형질 교정을 유도하는 획기적인 기술이다. 본 연구에서는 세포벽으로 인해 형질교정율이 동물 시스템에 비하여 상대적으로 낮은 식물세포에 적용시켜 효율을 높이기 위한 조건을 확립하고자 함을 연구목적으로 하였다. 타겟 유전자인 질소환원효소(Nitrate reductase)에 맞춤 제작된 3세대 유전자가위 RGEN (RNA-guided Engineered nuclease)을 이용하여 페튜니아의 원형질체 수준에서 고효율의 형질교정을 유도시키는 조건을 조사하였다. 종자로부터 기내에서 자란 폐튜니아의 어린 잎을 사용하여 cellulose, viscozyme, pectinEX이 포함된 혼합 효소액을 처리한 후 원형질체의 분리를 유도하였다. 예비 실험으로 PEG와 형질전환에 사용된 플라스미드 DNA인 pBI1221-GFP의 농도를 조절하여 원형질체에 도입한 결과, PEG의 농도가 40%이고 Plasmid DNA의 농도를 50ug을 이용하였을 때, 30% 이상의 가장 높은 유전자 도입 효율을 보이는 것을 확인하였다. 동일한 조건으로 페튜니아 NR 유전자에 맞춤 제작된 CRISPR/Cas9을 원형질체에 도입하여 세포배양을 실시한 후 배양세포로부터 DNA를 추출하여 mid-seq을 통한 변이체 발생 비율을 확인한 결과 최대 12%까지 타겟 유전자의 교정이 유도됨을 확인할 수 있었다. 본 연구에서 확립한 조건을 바탕으로 다른 가지과 작물의 다양한 선별 유전자에 적용시켜 목적 형질의 교정을 유도할 수 있는 새로운 작물 육종기술로 본 유전체 편집기술이 이용되도록 그 기반을 확립할 것이다.

Myo-inositol-1,2,3,4,5,6-hexakisphosphate (InsP6), also known as phytic acid, accumulates in large quantities in plant seeds, serving as a phosphorus reservoir, but is an animal antinutrient and an important source of water pollution. Here we report that Gle1 (GLFG lethal 1) in conjunction with InsP6 functions as an activator of the ATPase/RNA helicase LOS4 (Low expression of osmotically responsive genes 4), which is involved in mRNA export in plants, supporting the Gle1-InsP6-Dbp5 (LOS4 homolog) paradigm proposed in yeast. Interestingly, plant Gle1 proteins have modifications in several key residues of the InsP6-binding pocket, which reduce the basicity of the surface charge. Arabidopsis Gle1 variants containing mutations that increase the basic charge of the InsP6-binding surface show increased sensitivity to InsP6 concentrations for the stimulation of LOS4 ATPase activity in vitro. Expression of the Gle1 variants with enhanced InsP6 sensitivity rescues the mRNA export defect of the ipk1 (inositol 1,3,4,5,6-pentakisphosphate 2-kinase) InsP6-deficient mutant, and furthermore, significantly improves vegetative growth, seed yield, and seed performance of the mutant. These results suggest that Gle1 is an important factor responsible for mediating InsP6 functions in plant growth and reproduction, and that Gle1 variants with increased InsP6 sensitivity may be useful for engineering high-yielding low-phytate crops.

A wheat mutant of low-molecular-weight glutenin (LMW-GS) “Gunji-2” at Glu-B3 locus was derived among the double haploid lines. Gunji-2 was derived from F1 plants of Keumkang and Olgeuru crosses using the wheat × maize system according to the procedures of Inagaki and Mujeeb-Kazi at International Maize and Wheat Improvement Center (CIMMYT). Deletion of Glu-B3 LMW-GS proteins was found by allele specific DNA marker, one dimensional SDS-PAGE and two dimensional gel electrophoresis (2-DGE). Tandom mass spectrometry (MS/MS) was used to obtain direct evidence of LMW-GS deletion. In addition, we examined the basic agronomic traits, protein content, dough properties of mixing and bread loaf volumeof Gunji-2 and parental wheat cultivars grown for two years. This mutant will represent a valuable resource in quality test for specific allele or gene at Glu-B3 locus.

Plant breeding requires genetic diversity of useful traits for crop improvement. EMS-induced mutation is practiced to generate mutations at loci regulating economically important traits and/or to knock out the genes to elucidate their functions. The present study was aimed to induce mutations in a Korean local land race Capsicum annuum ‘Yuwol-cho’. This accession is pungent and also has advantage to mature early. A total of about 1,500 M2 families were screened and three non-pungent mutants were identified and crossed with wild type ‘Yuwol-cho’. After phenotyping of F2 population for pungency, MutMap approach will be used to identify the genes controlling the pungency in mutants. In addition to this, another C. annuum accession “Micro-Pep” was used to develop a mutant population. Micro-Pep is a small, pungent pepper generally used as ornamental purpose. Having compact growth habit, and small size, it has advantage to handle and utilize easily in mutation study and molecular research. On the basis of preliminary experiment 1.3% of mutagen was used for treatment of pepper seeds and 30% less germination percentage was observed in EMS treated seeds in comparison to control seeds. A total of 4,674 M1 plants are grown under greenhouse condition and M2 population will be studied for characterization of phenotypic variation including fruit color and pungency. Newly constructed mutant populations will be valuable assets for identification of functional genes and molecular breeding of pepper.