The selectable marker-free rice plants containing mcry1Ac insecticidal gene isolated from Bacillus thuringiensis (Bt) were generated using a non-selection approach by Agrobacterium tumefaciens-mediated transformation. The nutritional composition of two lines of transgenic rice plants (RTB5 and RTB11) was compared with that of its non-transgenic counterpart. The results showed that, except for small differences in dietary fiber and some minerals, there was no significant difference between transgenic rice and conventional counterpart variety with respect to their nutrient composition. Most of measured levels of nutrients were within the range of values reported for other commercial cultivars, showing substantial equivalency. Therefore, the insertion of transgenes did not affect the nutritional composition of transgenic RTB5 and RTB11 rice grains.

The Asteraceae/Compositae family is one of the biggest families in flowering plants and has more than 23000 species including the economically important lettuce, sunflower, and chicory as well as the agronomic weeds. With its significance and the progress in sequencing technology, its species have been subjected to the genome sequencing project worldwide. Although chrysanthemum is an important plant in the floricultural industry, however, it has been less studied at the level of genomics, compared with other species in the Asteraceae. There were only several reports on comparative analysis of transcriptome for chrysanthemum. Actually, the genome of Chrysanthemum species is known to be gigantic and complex with diverse status ranging from diploid to decaploid. Since the cultivated and commercial chrysanthemum exhibits hexaploid genome, we decided to select the diploid species with smaller genome as a material for reference genome sequencing. Thus, we launched a genome sequencing project with C. boreale which was previously reported to be diploid by cytogenetic analysis. We constructed sequencing libraries with insert size 300bp and 500bp and sequenced them from the paired end in 100bp read length with Illumina’s HiSeq platform. After quality checking, we preprocessed raw reads by removing duplicated reads and trimming reads with low quality value. Kmer frequency analysis with the cleaned reads showed that the genome is heterozygous, highly repetitive and gigantic, ranging from 2.9Gb to 5.8Gb. The cleaned reads were further subjected to error correction and primary assembly with SOAPdenovo2. Here, we’ll report the result of Kmer frequency analysis and genome assembly.

억새(Miscanthus spp.)는 화본과 중 광합성 효율이 높은 C4 식물군에 속하는 식물로 한국, 일본, 중국 등 동아시아가 원산지인 대표적인 바이오에너지 원료작물이다. 억새는 주로 지하경을 이용하여 번식하여 왔으나 지하경을 이용한 번식은 유전형은 유지할 수 있으나 우량 품종 개발에 불리하다는 문제점이 있다. 조직배양기술은 유용 자원을 이용한 돌연변이 육종과 형질전환 기술을 이용한 신품종 육종을 위한 기반기술로 활용 될 수 있어 바이오매스의 확보측면에서 유용하다. 국내 유망 바이오매스 자원인 억새의 신품종 육성을 위해서는 캘러스 유도 및 식물체 재분화에 효율적인 조직배양 기술을 확립하는 것이 중요하다. 본 연구에서는 농촌진흥청에서 선발한 ‘우람’ 억새(Miscanthus sacchariflorus cv. Wooram)의 미성숙 화기를 이용한 안정적인 캘러스 유도 및 식물체 재분화 조건을 확립하여 신품종 육종을 위한 기초자료를 확보하고자 하였다. 식물재료는 국립식량과학원 바이오에너지작물연구소 내의 억새 재배포장에서 미성숙 화기가 5 mm 이하로 분화한 개체만 채취하여 사용하였으며, 수집한 재료는 70% EtOH로 2분, 0.45% NaOCl으로 20분간 표면 살균하여 배지에 치상하였다. 미성숙화기로부터 캘러스 유도율을 조사한 결과, MS배지에 생장조절제인 3 mg L-1 2,4-D를 첨가한 처리가 캘러스 유도율 93.3 %로 가장 높게 나타났고 3 mg L-1 2,4-D + 0.1 mg L-1 BA를 혼합 처리한 배지에서도 86.6 %의 캘러스 유도율을 나타냈다. 이후 캘러스로부터 식물체 재생 실험에서 3 mg L-1 2,4-D에 0.1 mg L-1 BA를 혼합 처리한 배지에서 유도된 캘러스가 3 mg L-1 2,4-D 처리 배지에서 유도된 캘러스보다 식물체 재분화율이 높게 나타났다. 캘러스 유도에는 최종적으로 3 mg L-1 2,4-D + 0.1 mg L-1 BA 처리 배지가 가장 효과적인 것으로 나타났다. 식물체 재분화를 위한 최적 생장조절제 농도에 대한 실험에서는 5 mg L-1 BA + 0.1 mg L-1 NAA 배지에서 재분화율 86.6 %로 가장 효과적이었다.

In the facultative long-day (LD) plant Arabidopsis thaliana, FLAVIN-BINDING, KELCH REPEAT, F-BOX1 (FKF1) is activated by blue light and promotes flowering through the transcriptional and post-transcriptional regulation of CONSTANS under inductive LD conditions. By contrast, the facultative short day (SD) plant rice (Oryza sativa) flowers early under inductive SD and late under non-inductive LD conditions; the regulatory function of OsFKF1 remains elusive. Here we show that osfkf1 mutants flower late under SD, LD, and natural LD conditions. Transcriptional analysis revealed that OsFKF1 up-regulates expression of the floral activator Ehd2 and down-regulates expression of the floral repressor Ghd7; these regulators up- and down-regulate Ehd1 expression, respectively. Moreover, OsFKF1 can upregulate Ehd1 expression under blue light treatment, without affecting the expression of Ehd2 and Ghd7. In contrast to the LD-specific floral activator Arabidopsis FKF1, OsFKF1 likely acts as an autonomous floral activator because it promotes flowering independent of photoperiod, probably via its distinct roles in controlling expression of rice-specific genes including Ehd2, Ghd7, and Ehd1. Like Arabidopsis FKF1, which interacts with GI and CDF1, OsFKF1 also interacts with OsGI and OsCDF1 (also termed OsDOF12). Thus, we have identified similar and distinct roles of FKF1 in Arabidopsis and rice.

Platycodon grandiflorum is a species of herbaceous flowering perennial plant of the family Campanulaceae. The major ingredients are platycosides, terpenoid saponins. It contains 1-4 % of the dry weight and there are about 20 types of platycosides. Among them, platycodin D have various pharmacological effects on cough and cold. Platycosides are synthesized from oleanane by mevalonic acid pathway and cytochrome P450s and UGTs are important enzymes in the saponin biosynthesis. UGT is glucose transfer enzyme and act on the final step of the secondary metabolite biosynthesis. In this study, we tried to identify UGT genes involved in saponin biosynthetic pathway from the various tissues of P. grandiflorum and non germinated seeds using RNA-seq analysis. The sequencing was performed using Illumina Hi-Seq platform after cDNA library preparation. The produced reads were assembled using CLC Genomics Workbench software (CLC Bio, Inc.). We obtained 122,663 contigs and found 137 putative UGT genes. The phylogenetic relationship was analyzed and putative genes related to platicoside biosysthesis were selected and cloned for further analysis.

Platycodon grandiflorum is a perennial herbal plant belongs to Campanulaceae family. It has very important genetic value as a major plant in Asterids order. The major ingredients are platycosides, terpenoid saponins. In Korean industrial plants market, it was produced 5,633 tons in 2013, and the total amount of production was less than only five species, omija, ginger, raspberry, yam and deodeok. P. grandiflorum is called ‘Gilgyung’ and is used as a fresh vegetable and an ornamental plant. Nowadays, various components of P. grandiflorum were already published. But, genetic research is in the starting stage. In this study, 11 cultivars; 1. MariesⅡ, 2. Hakone double white, 3. Hakone double blue, 4. Fuji white, 5. Fuji pink, 6. Fuji blue, 7. Astra white, 8. Astra pink, 9. Astra blue, 10. Astrasemi double blue, 11. Jangback, were analyzed using 60 Operon Universal RAPD primers. The results were phylogenetically analyzed and related to the morphological characteristics of the cultivars.

Platycodon grandiflorum is a herbal flowering perennial plant belongs to Campanulaceae family. The saponins derived from P. grandiflorum were termed platycosides and platycodin D, which is the most abundant saponin in the plant and pharmacologically active component, was intensively studied. Platycodin D is synthesized from triterpenoids by several enzymes including cytochrome P450. Cytochrome P450 is known to exist in superfamily in plant kingdom and essential roles in saponin biosynthetic pathway by hydroxylation or oxidation of triterpene skeletons. However, the key genes of P450 involved in biosynthesis of saponin was not identified because of its low conservation rate in amino acid sequence level among plant species and gene superfamilies. Recently, next generation sequencing (NGS) technology is rapidly developed as a method to discover target genes. In this study, we tried to identify P450 genes involved in saponin biosynthetic pathway from the various tissues of P. grandiflorum using RNA-seq analysis. The sequencing was performed using Illumina Hi-Seq platform after cDNA library preparation. The produced reads were assembled using CLC Genomics Workbench software (CLC Bio, Inc.). We obteined 122,663 contigs and found out 191 putative P450 genes. The phylogenetic relationship was analyzed and putative genes related to platicoside biosysthesis were selected and cloned for further analysis.

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.

Carotenoid isomerase (CRTISO) catalyzes the isomerization of prolycopene to all-trans-lycopene in the carotenoid biosynthetic pathway. We isolated two full-length cDNA gene, CuCRTISO and CuCRTISO-like, from Citrus unshiu. To confirm whether these two genes have the function of the carotenoid isomerase, The full-length cDNA of CuCRTISO and CuCRTISO-like gene, respectively, were fused with 35S promoter and NOS terminal region and then transformed into tomato CRTISO mutant, Tangerine, which shows orange fruit due to lack of carotenoid isomerase activity. The mature fruit color of the transgenic line expressing CuCRTISO gene changed from orange to red, which was similar to the fruit color of the tomato “Money Maker”. We also carried out HPLC analysis to detect all-trans lycopene, which is produced from prolycopene by carotenoid isomerase. In the transgenic line expressing CuCRTISO the all trans lycopene was detected from mature fruit but in the tangerine mutant several prolycopenes were detected from it. On the other hand, the transgenic line expression of CuCRTISO gene retained the orange-color fruit at the mature stage as Tangerine mutant. These studies indicate that the CuCRTISO gene has a function of carotenoid isomerase and also plays a role of it in other plant species, and that the CuCRTISO-like gene might be not enough to produce the all trans lycopene or has a another unknown function(s).

MLO is a unique gene family which is identified in plant and carries out abiotic and biotic stress responses in various plants. The understanding on the roles and functional diversity of this family is quite limited in rice, a model crop plant. Rice genome has 12 potential MLO family members. To do systematic functional assignment of MLO family in rice, we performed phylogenomic analysis of integrating meta-expression data based on public sources of microarray data or RT-PCR data into the phylogenic tree. As a result, we identified 12 MLO genes carrying various tissue-preferred expression patterns such as leaf, root, pollen, and ubiquitous expression, suggesting functional diversity in terms of anatomy or development. RT-PCR analysis confirmed, integrated transcriptome data were used to estimate the functional redundancy or specificity among MLO family: MLO12 showed mature pollen preferred expression; MLO4, root tip; MLO10, overall root except root tip; MLO8, leaf; MLO2 and MLO9 showed redundant expression in overall tissues except root. Also, abiotic stress meta-expression data and RT-PCR performance suggested the functional association of 5 MLO and 6 MLO genes with heat and cold stress, respectively. Our analysis will provide basic information to study diverse developmental or physiological phenomena mediated by MLO family in rice, a major model crop plant.

FT is one of the major floral activator in photoperiod-dependent flowering pathway. To understand the role of FT homologs in flowering time control of short-day plant soybean, we identified ten soybean FT genes and named GmFTs. Phylogenetic analysis revealed that ten GmFT genes were further categorized into three subclades. Gene expression analysis showed that the most GmFT genes are mainly expressed in leaves. The expression of GmFT2a, GmFT2b, GmFT5a, and GmFT6 was strongly induced under the floral inductive short-day condition, but GmFT4 exhibited opposite expression pattern compared to those of GmFT2a, GmFT2b, GmFT5a, and GmFT6. To understand roles of GmFT genes in flowering, we generated Arabidopsis transgenic plant overexpressing GmFT genes. Both 35S:GmFT2a and 35S:GmFT5a transgenic plants showed extremely early flowering. In contrast, overexpression of GmFT4 delayed flowering of transgenic plants compared to wild type Arabidopsis. The results indicated that GmFT2a and GmFT5a might function as floral activators, while GmFT4 has an opposite function in soybean flowering. Moreover, domain swapping approaches between GmFT2a and GmFT4 revealed that the substitution of the segment B region alone, which is located in 4th exon, was sufficient to change the function of GmFT2a to floral repressor and GmFT4 to floral activator. The results suggested that soybean FT homologs have been functionally diversified during evolution and might play different roles in photoperiod-dependent flowering of soybean.

To develop transgenic forage crops with enhanced tolerance to abiotic stress, we introduced an alfalfa Hsp23 gene expression vector construct through Agrobacterium-mediated transformation. Integration and expression of the transgene were confirmed by PCR, northern blot, and western blot analyses. Under normal growth conditions, there was no significant difference in the growth of the transgenic plants and the non-transgenic controls. However, when exposed to various stresses such as salt or arsenic, transgenic plants showed a significantly lower accumulation of hydrogen peroxide and thiobarbituric acid reactive substances than control plants. The reduced accumulation of thiobarbituric acid reactive substances indicates that the transgenic plants possessed a more efficient reactive oxygen species-scavenging system. We speculate that the high levels of MsHsp23 proteins in the transgenic plants protect leaves from oxidative damage through chaperon and antioxidant activities. These results suggest that MsHsp23 confers abiotic stress tolerance in transgenic forage crops and may be useful in developing stress tolerance in other crops.

청원 소재 음나무(Kalopanax septemlobus)의 미숙종자에서 캘러스를 유도하여 15개의 배발생 캘러스를 얻었다. 증식된 배발생 캘러스를 재료로 체세포배를 유도하여 15개체의 기내 식물체를 얻었다. 체세포배 유도는 1/2 MS배지에 0.1 mg/L abscisic acid (ABA), 7% polyethylene glycol (PEG), 0.02% activated charcoal, 3% sucrose를 첨가하고, 0.5% gelrite로 경화하여 사용하였다. 식물 재생용 배지는 배지는 1/2MS배지에 2% sucrose, 0.3% gelrite로 하였고, 발아 촉진을 위해 GA3 1.0 mg/L 처리 혹은 기본배지를 사용하였다. 15개체의 체세포배 발생 빈도는 다르게 나타났고, 재생된 식물체의 GA3 효과는 크지 않았다. 유전적 다양성을 조사하기 위하여 ISSR (Inter-Simple Sequence Repeats) 표지자 분석을 실시하였다. 5개의 ISSR 프라이머에서 증폭산물을 관찰하였고, 유전적 다양성을 나타내는 P (Percentage of polymorphic loci)값과 S.I. (Shannon’s information index)를 조사하였다. ISSR 마커를 이용하여 재분화 식물체의 유전적 안정성을 분석한 결과, 체세포 유래 재분화된 식물체의 개체간에 유전적 구조가 균일하며, 유전적 변이는 관찰되지 않았다.

어유에서 추출한 Omega-3 long-chain polyunsaturated fatty acids (LC-PUFA), 특히, EPA (20:5)와 DHA (22:6)는 심혈관계질환을 예방하는데 중요한 역할과 함께 대사성 증후군 또는 비만 발병과 관련있다고 알려져 있다. 동물 모델을 이용해 고지방식이 섭취 후 어유 추출 오일과 식물성 오일 투여로 혈중 지질 농도 감소 효과를 비교하고 작용 기전을 확인함으로써 고지혈증을 포함한 심혈관계질환의 예후인자를 알아보고자 한다. 42마리의 C57BL6J 마우스를 이용해 정상식이군 (18마리)과 고지방식이군 (24마리)으로 나눈 후 정상식이군 3개 그룹 (대조군, 어유 오일, 식물성 오일)으로 설정하고 고지방식이군도 3개 그룹 (대조군, 어유 오일, 식물성 오일) 각각으로 나누어 실험을 진행했다. 고지방식이군은 4주 단위 투여 로 비만을 유도한 후 체중 20g 당 100μl의 식물성 오일과 어유 추출 오일을 각각 투여하여 각 그룹 별로 매일 1회 씩 털 색깔, 몸 전체 모양, 털 빠짐 등 일반증상을 관찰하고 사망동물이나 빈사 상태를 확인했다. 오일 투여 직전과 투여 후 1, 3, 7일에 체중변화를 측정하고 10주 후 마취시켜 부검 후 외관검사 실시와 육안 소견을 관찰하고 장기를 적출하여 효소항체검사법 (enzyme-linked immunosorbent assay test, ELISA)를 통해 지방 활성에 관련된 혈중 지질 농도를 측정했다. 고지방식이 섭취 4주 후 정상식이군 (p=0.56)에 비해 고지방식이군 (p=0.04)에서 체중변화가 나타났고 고지방식이군에서 어유 오일군 (41.27±7.0)에 비해 식물성 오일군 (45.37±6.45)에서 체중감소가 적었지만 물만 먹인 대조군 (48.87±1.0)에 비해 두 그룹 모두 체중이 감소하는 효과가 있었고 (p=0.04), 10주 투여기간동안 오일 투여군에서 체중 감소가 나타났다. 부검 후 적출한 장기에서 관찰된 혈중 지질 농도 수치 역시 정상식이군에 비해 고지방식이군에서 감소하는 것으로 보였지만, 고지방식이군 중 어유 오일 그룹과 식물성 오일 그룹에서 혈중 지질 농도 수치 감소가 거의 비슷한 수준으로 나타나는 것을 알 수 있었고 추후 동물모델을 추가 시험함으로써 확인하는 것이 필요할 것으로 보인다. 식물성 오일 섭취로 혈중 지질 수치가 감소함을 관찰함으로써 향후 혈관질환의 진단 및 예후인자로 이용할 수 있을 것으로 여겨진다.

Small heat shock proteins (Hsps) are one of most conserved molecular chaperones that protect stress-inducible denaturation of substrates in living organisms. Small Hsps consist of a large subfamily categorized by subcellular localization ranging in size from 12 to 40 kDa. Here, we identified and characterized a small Hsp 16.9 gene (EsHsp16.9) from Siberian wild rye (Elymus sibiricus L.). EsHsp16.9 is a 456-bp cDNA with an open reading frame predicted to encode a 151-amino acid protein. It possesses a conserved ɑ-crystallin domain, which is a unique domain for small Hsps; shares high sequence similarity with cytosolic class I small Hsps among the small Hsp subfamily in Arabidopsis; and is close (96% similarity) to small Hsp in wheat. Northern blot analysis showed that EsHsp16.9 transcripts were enhanced by heat, drought, arsenate, methyl viologen, and H2O2 treatments. Moreover, we expressed and purified recombinant EsHsp16.9 proteins in Escherichia coli to confirm its activity as a molecular chaperone. We found that recombinant EsHsp16.9 exhibits effective molecular chaperone activity, as determined by inhibition of thermal aggregation of malate dehydrogenase (MDH), which is broadly used as a model substrate.

아열대 및 온대지역에서 주로 발생하는 애멸구는 5월∼8월 사이 편서풍을 타고 우리나라에 비례하며 벼 바이러스병인 줄무늬잎마름병을 매개한다. 줄무늬잎마름병은 벼의 수량감소와 미질을 떨어뜨리는 주요 병해 중 하나이다. 애멸구와 같은 멸구류는 대부분 살충제에 의해 방제를 하고 있으나 이러한 약제의 계속된 사용은 환경오염과 약제저항성 개체의 발생을 유발시킬 수 있는 큰 단점이 있다. 이와 같이 벼에 심각한 피해를 입히는 병해충에 대한 저항성 품종 육성은 병해충을 방제하는 가장 경제적이고 효과적인 방법으로 알려져 있다. 줄무늬잎마름병의 저항성 관련 유전자는 ‘Modan’에서 유래한 Stv-bi, 일본 밭벼 또는 열대 자포니카 품종에서 유래한 Stv-a, Stv-b가 알려져 있으며, 국내에서는 주로 Stv-bi가 도입된 저항성 품종들이 육성되고 있다. 단일 유전자를 이용한 저항성 품종육성은 새로운 바이러스의 등장에 취약하므로 저항성 유전자원의 다양화와 새로운 유전자의 탐색이 필요하다. 본 실험 에서는 Stv-a 와 Stv-b가 보고된 USA 품종인 Zenith의 형질을 일품에 도입하여 180계통의 F2 집단을 육성하여 생물검정과 함께 QTL분석에 이용하였다. 줄무늬 잎마름병에 대한 Zenith에 대한 QTL을 탐색한 결과, 11번 염색체에서 LOD 11.9, 설명가능한 표현형 변이 27%인 QTL이 확인되었다. 줄무늬 잎마름병 연관 QTL 및 이와 관련된 marker는 향후 줄무늬잎마름병에 대한 새로운 저항성 유전자인 Stv-b를 보유하는 품종육성을 위한 MAS 체계 확립에 이용할 것이다.

To understand the molecular mechanism of leaf morphogenesis in rice, ethylmethane sulfonate (EMS) treated Ilpoom mutant line with semi-narrow and adaxially rolled leaf phenotype was identified. The leaf rolling character is said to be more advantageous under high temperature and heat stress, and play as one of the defensive mechanisms. The F1 plants, generated from a cross of Ilpoom and mutant, showed normal phenotype. Genetic analysis of its F2 population suggested that the mutation was controlled by a single recessive gene with segregation ratio of 3:1. Using F2 mapping population derived from a cross of Ilpoom mutant and Milyang23, each chromosomes were screened with STS markers by the bulked segregant analysis (BSA) method. The candidate region was detected to a long arm of chromosome 1 near the centromeric region. Fine mapping of the locus is currently conducted. Moreover, other morphological characterizations of the mutant plants were identified. Cytological analysis of the leaf suggested that deformation of the bulliform cells led to the smaller size and less number of the bulliform cells, and caused leaf rolling trait.

Although it is known that the composition of HMW-GSs and LMW-GSs are important factor for end-product quality as bread, noodle and cookie, it is still not clear which HMW-GSs and LMW-GSs confer specific processing properties. In this study, to investigate distinctive glutenin proteins and expression level for characteristic processing properties, we carried out qualitative and quantitative analysis of gluetenin protein in noodle and bread wheat cultivars by two-dimensional electrophoresis. Unexpectedly, five LMW-GS spots were found to be expressed at a common position in all cultivars and these spots may play something in glutenin biosynthesis. Also we found LMS-GS spots to distinguish Korean wheat cultivars mostly used as noodle and western bread wheat cultivars. These spots may contribute to characteristic processing properties. The 2DE results for each cultivar will be used as reference map or protein marker discriminating wheat cultivars, wheat and rice, imported and Korean flour. For quantitative analysis of gluetenin, we calculated relative expression level of the HMW-GS, LMW-GS and HMW-GS/ LMW-GS ratio in each cultivar by 2DE. The results presented in this study provide new insight into relation of specific glutenin proteins and end-use quality and will be useful to choose elite breeding line for improvement of wheat flour quality.

Soybean is a crop of importance economically and nutritionally in many parts of the world. Thanks to many new genes brought from genomic research, It is possible to introduce various candidate genes through genetic transformation to see the performance of the genes in field. In our lab, soybean transformations have been tried for last 10 years to probe the possibility of traits improvement by transformation of new gene into soybean. For this purpose, three different genes were transformed into Korean soybean variety, Kwangan. First, the gene that controls early flowering of plant was transformed into Kwangan. Second, a candidate gene for soybean mosaic virus (SMV) resistance was transformed to produce transgenic plants. Third, another candidate gene for drought tolerance was transformed. All the transgenic plants from three genes transformation were produced for their gene insertion and their expression using PCR, qRT-PCR. Further analysis including harvesting seeds is currently undertaken.

Although it is well known that low-molecular-weight glutenin subunits (LMW-GS) affects bread and noodle processing quality, the function of specific LMW-GS proteins mostly remain unclear. It is important to find a corresponding gene for a specific LMW-GS protein in order to understand the function of the specific LMW-GS protein. The objective of this study was to identify LMW-GS genes and haplotypes using well known Glu-A3, Glu-B3 and Glu-D3 gene specific primers and to interlink their protein products by proteomic approaches in a wheat variety. A total of 36 LMW-GS genes and pseudo-genes were amplified including 11 Glu-3 gene haplotypes, designated as GluA3-13K and GluA3-22K (pseudogene) at Glu-A3 loci, GluB3-33K and GluB3-43K at Glu-B3 loci and GluD3-11K, GluD3-21K, GluD3-31K, GluD3-42K, GluD3-5K, GluD3-6K and GluD3-393K (pseudogene) at Glu-D3 loci. To determine the relationship between gene haplotypes and their protein products (to identify the corresponding LMW-GS proteins), we conducted N-terminal amino acid sequencing and tandem mass spectrometry (MS/MS) analysis of the 17 LMW-GS spots separated by 2-DGE. Successfully, LMW-GS proteins of the Glu-3 gene haplotypes except pseudo-genes mentioned above were identified. This is the first report on comprehensive characterization of LMW-GS genes and their corresponding proteins and establishment of specific correspondence between each other in a single wheat cultivar. Our approach will be useful to understand the molecular basis of the LMW-GS and to study their contribution to the end-use quality of flour.