Grain weight (GW) or grain size is the most important target not only as a major component of grain yield, but also associated with the cooking quality in rice breeding program. In a previous study, two loci for grain weight, gw8 and gw9, have been fine map to about 306.4kb and 37.4kb, respectively, in backcross populations derived from a cross between the Korean japonica cultivar Hwaseongbyeo and Oryza rufipigon (IRGC 105491). Both O.rufipogon alleles increased GW in the Hwaseongbyeo background despite the fact that O. rufipogon was the small-seeded parent. To test the effect of pyramiding the QTLs for grain weight, an F2 population consisted of 226 individuals was developed derived from the cross between the two QTLs (gw8 and gw9), and used MAS to select nearly isgenic lines for two QTLs and a pyramiding line. Molecular genotyping of F2 population revealed the existence of interaction between gw8 and gw9. Moreover, the interaction was also confirmed by phenotypic analysis of four QTL-NILs. The results suggest that epistasis interaction is important genetic basis in determining yield traits in rice.

Mongolia is known as the livestock country for 800 years from the period of Chinggis Khaan Empire. The livestock has been raised without corn, a high quality feed. Hence, the meat of Mongolia has not been exported. In 2004, International Corn Foundation (ICF) and KNU in Korea initiated corn breeding program for Mongolia with Mongolian Academy of Science and Agricultural University of Mongolia. Since that time approximately 2000 genotypes were planted in Agricultural Research Station in Darkhan, Mongolia. Within 4years of research we have succeeded in forming Mongolian corn population (MCP), which can be used to breed locally adapted varieties in different zones. MCP characterizes early maturity, cold and drought tolerances. In 2008, 160 S1 were planted and 16 lines were selected. For further inbreeding and population improvement we planted all 16 lines in KNU vinyl house at Kunwi in late September. F1 crosses and advanced S3 lines will be tested in 2009 first time for hybrid development in Mongolia.

The puroindoline genes (Pina and Pinb) losely linked on the short arm of the 5D chromosome (Hardness locus (Ha)) of hexaploid wheat (Triticum aestivum L.) are of great importance to wheat and product quality. In this study, the barley cultivar Harrington was transformed with wheat gene pina and pinb by particle bombardment and performed in order to develop softer barley lines. The result, an efficiency of 3.9% of the bombarded calli regenerated transgenic plants. Each transgenic event was obtained from the callus of a different immature embryo. They showed the multiple shoots compared with control (no bombardment). Two plants from T0 lines were PCR positive for only the 'soft type' pinb transgene, 10 plants were positive for the 'soft type' pina transgene only, whereas they did not show both 'soft type' pina and pinb transgene. These results supply more evidence to support the hypothesis that puroindolines are potential genes for barley grain hardness and used to be suitable target for the production of transgenic barley plants to their availability biolistic transformation system.

This experiment was carried out to investigate the relationship between grain shapes and ecotypes in Korean native weedy rice. The 111 lines of Korean weedy rice were classified into japonica (91 lines) and indica (20 lines) by RAPD analysis, which was reported in 2005 (Chung and Ahn). The characteristics of grain shape such as grain length, width, thickness, 1,000 grain weight, the ratio of length to width (L/W), the ratio of length to thickness (L/T), and the ratio of width to thickness (W/T) were measured in each ecotype, and their statistical distributions were compared by t test. The mentioned 7 characteristics were significantly different between japonica and indica weedy rice groups, but these could not classify weedy rice accessions into two groups without exceptions according to the classification by RAPD analysis. Out of variables concerning grain shape, the L/W was the highest distinctive to separate ecotypes. A critical point of L/W to divide ecotypes was calculated from the mid point between upper critical limit of japonica and lower critical limit of indica in confidence interval at α=0.05. At critical point of L/W = 2.138, decision error was 2.2% in japonica group, and 5.0% in indica group. Conclusively, the classification of ecotypes by L/W of grain made almost (under 5% error) same result with that by RAPD markers in Korean native weedy rice.

We isolated low temperature inducible genes using suppression subtractive hybridization (SSH) method and were able to obtain to clone MLT7 gene encoding peroxiredoxin and aminotransferase. The full-length cDNA of MLT7 is 1,049 bp with an open reading frame (ORF) consisting of 261 amino acid (aa). Genomic southern blot confirmed that mungbean genome has two copies of MLT7 gene. Northern blot analysis was also carried out for the gene expression during ABA, NaCl, drought, wounding and H2O2 stresses. The expression of MLT7 gene significantly decreased by ABA, NaCl and drought stress, but wounding and H2O2 stress significantly induced MLT7 gene expression. Especially, H2O2 strongly induced the MLT7 gene expression. The expression of MLT7 gene during low temperature stress started to increase in 3 h after treatment, and than slightly decreased and again increased at 24 h. Using GFP fusion vector, GFP-MLT7 was targeted both to mitochondria and chloroplast. However, it was mostly targeted to mitochondria and partially targeted to chloroplast. For the functional analysis of MLT7, MLT7 recombinant protein was heterologously expressed in E. coli. The MLT7 recombinant cells showed enhanced antioxidant activity compared to that of vector control cells.

Xanthomonas oryzae pv. oryzae에 의해 유발되는 벼흰잎마름병은 우리나라에서는 남부평야지를 중심으로 발생하여 수량 감수는 물론이고 등숙 및 천립중을 저하시키고 미질을 떨어뜨린다.또한, 벼흰잎마름병균은 도관부에서 세균이 증식되어 발생하기 때문에 약제방제 효율이 매우 낮으므로 본 병의 효과적인 방제는 저항성 품종의 재배이다. 본 연구는 벼흰잎마름저항성 유전자의 유무 및 종류가 잎에서 벼흰잎마름병균의 증식 및 병반장 신장에 미치는 영향을 조사하기 위하여 실시하였다.벼흰잎마름병균을 접종 하여14일후에 세균밀도를 측정한 결과 저항성 유전자가 없는 Toyonishiki와 Milyang23호에서는 세균밀도가 5×105 ~ 2.8×106cfu/cm2이고 병반장은 8.7 ~ 18.7cm로 나타났다. Xa1 유전자를 갖는 계통은 병반장이 0.8cm인 균주의 세균 밀도는 3.1×104cfu/cm2이었으나 5cm이상에서는 대부분의 균주가 105cfu/cm2이었으며, Xa3 유전자를 갖는 계통은 저항성 반응에서는 대부분의 세균밀도가 103 ~ 104cfu/cm2 , 감수성에서는105cfu/cm2로 병반장이 길어질수록 병원세균의 밀도가 증가하는 경향이었다. xa5, Xa7유전자를 갖는 계통은 저항성 반응에서는 103 ~ 105cfu/cm2의 세균밀도로 나타났으며 감수성반응에서는 5×105 ~ 2.8×106cfu/cm2의 세균밀도로 다른 저항성 품종에 비하여 높은 세균밀도가 존재함에도 저항성 반응을 나타내는 경향이었다.

We isolated wound-inducible genes using suppression subtractive hybridization (SSH) method and were able to obtain to clone W3 gene encoding dnaJ like protein. The full-length cDNA of W3 is 689 bp with an open reading frame (ORF) consisting of 163 amino acid (aa). Genomic southern blot confirmed that soybean genome has two copies of W3 gene. Northern blot analysis was also carried out for the gene expression during heat, NaCl, drought, wounding stresses. The expression of W3 gene specifically induced by heat, NaCl, wounding and drought stress. Using GFP fusion vector, W13-GFP was targeted both to nucleus. For the functional analysis of W3, His-tagged W3 recombinant protein was heterologously expressed in E. coli. The W3 recombinant cells showed enhanced heat tolerance compared to that of vector control cells. We suggest that dnaJ-like W3 protein function as molecular chaperone in the nucleus of the plant cell during various stresses.

Photorespiration reduces carbon fixation rate, but is essential process in plant. Photorespiration involves reactions in chloroplasts, peroxisomes, and mitochondria. In photorepiratory peroxisome, alanine glyoxylate aminotransferase (AGT) catalyzes alanine and glyoxylate into glycine and pyruvate. We isolated a low temperature-inducible cDNA encoding AGT from mungbean leaves. The full-length cDNA, designated as MLT9, contains an open reading frame of 1,203 nucleotides coding for a protein of 401 amino acids. Genomic DNA blotting showed that the mungbean genome has one copy of MLT9. MLT9 mRNA was induced not only by low temperature but also by drought stress, but ABA and NaCl did not induce RNA expression of MLT9. In mungbean, AGT activity was higher in the non-stressed leaves compared to the low-temperature treated leaves. Based on GFP/RFP targeting experiment, GFP-MLT9 fusion protein and SKL-RFP, a peroxisome marker, were colocalized to peroxisome in tobacco protoplasts. This suggests that peroxisomal MLT9 plays a role in photorespiratory metabolism in response to low temperature and drought stress.

IL-34 (NIL) developed by introgressing chromosomal segment substitution from an accession of Oryza minuta (2n=48, BBCC, Acc. No. 101141) into the O. sativa subsp. japonica cv. Hwaseongbyeo, showed significantly higher number of spikelets per panicle (SSP) than the recurrent parent Hwaseongbyeo. QTL analysis in F2 generation derived from the cross between IL-34 and Hwaseongbyeo revealed that ssp7, a QTL was located in the pericentromeric region of chromosome 7. The frequency distribution of spikelets per panicle followed 3:1 ratio for single locus segregation. The additive effect of the O. minuta allele at the QTL was 23 spikelets per panicle, and 43.6% of the phenotypic variance could be explained by the segregation of marker RM21596. To clarify whether ssp7 could be dissected genetically, we carried out fine-scale mapping with 3,700 F2 plants derived from the cross between IL-34 and Hwaseongbyeo using markers flanking spp7. 186 F2 plants having informative recombination breakpoints within the region flanked by two SSR markers RM500 and RM21615 were identified and used for fine mapping of ssp7. ssp7 was mapped between the SSR markers RM21596 and RM418 which was approximately 441kb in length based on the physical map of the region. Of great interests, the QTL region also had effects on primary branch number (PB), grains per panicle (SP) and grain yield (YD). These results are very useful for transferring or pyramiding ssp7 by molecular marker assistant selection in rice breeding programs.

We isolated two low temperature-inducible cDNAs designated as MLT5 and MLT31 from mungbean by subtractive suppression hybridization method. By rapid amplification of cDNA end technique, the full-length cDNAs designated MLT5 and MLT31 were obtained. The full-length cDNA of MLT5 contains an open reading frame of 324 nucleotides coding for 107 amino acids. The full-length cDNA of MLT31 contains an open reading frame of 444 nucleotides in length and capable of specifying a 16.5-kDa protein of 148 amino acids (aa) with an isoelectric point of 7.72. RNA expression of MLT5 was strongly induced by low temperature in the early time and also by wounding, NaCl and ABA. MLT31 mRNA was induced by NaCl and ABA but not by wounding and low temperature stress. MLT5 encodes a protein of unknown function, of which targeting site is predicted to be chloroplast membrane protein. To examine the localization of MLT5, MLT5-GFP was expressed in tobacco cells. It was shown that MLT5-GFP was localized to surface of chloroplast in tobacco cell. To examine the function of MLT31, MLT31 was expressed in Escherichia coli as His-fusion protein. Purified MLT31-His recombinant protein will be tested for ubiquitination activity in vitro. In addition, for the in vivo functional analysis of MLT31, MLT31 will be expressed in yeast ubc9 knock-out mutant. We propose that MLT5 and MLT31 play an important role in signal pathway of abiotic stress in plants.

생물자원의 유전적 다양성을 유지하기 위해서는 보유하고 있는 유전자원의 분석을 근거로 한 다양성확보가 필수적이다. 본 연구는 ICARDA에서 분양받은, 시리아등 87개 지역에서 수집한 야생보리(Horedeum spontaneum) 394점을 재료로 농업적형질 및 표현형을 조사하여 구분정리함으로서 정제된 유전자원의 보존과 이용효율성을 높이고자 연구를 실시하였다.질적형질은 총성을 포함한27개의 형질을 조사하였고, 양적형질은 간장을 포함한 12개의 형질을 조사하였다. 과성 및 찰성을 지닌 자원은 없었으나 탈립성이 강한 7점의 자원을 선발할 수 있었다.추후 철분등 종실의 영양성분 분석 및SSR마커를 이용한 유용관계의 분석이 이루어질 예정이며, 이들 결과는 야생보리를 이용한 기능성 보리 품종 육성에 이용될 것이다.

특정형질과 관련된 유전자 특이적 분자마커를 개발하는 과정은 육종의 효율을 높이는 매우 중요한 과정이나 기술적으로 근동질계통간 유전체 수준에서 다형성을 발굴하는 것은 다양한 데이터베이스 마이닝기법을 사용한다 하더라도 결과 예측이 불확실한 작업이다. 더욱이 근동질계통간 다형성이 발견되었다 하더라도 그것이 어떤 형질과 연관되어 있는지를 밝히는 것은 전혀 별개의 실험적 분석이 필요하다. 본 실험에서는 유전체수준에서 높은 유사성을 가지고 있는 자포니카 타입 벼들간의 교배조합을 대상으로 하여 다양한 형질이 분리하는 200여 계통을 대상으로 하여 전사조절유전자집단에서 발굴한 근동질계통간 유전자 특이적 분자마커를 이용하여 46개의 표현형과의 연관관계를 통계 분석하였다. 두 개의 서로 다른 벼 염기서열 데이터베이스가 사용되었으며 다양한 벼 염기서열 데이터마이닝 기법이 사용되었다. 본 실험을 통하여 특정 유전자와 밀접하게 연관된 부위의 다형성은 유전자 개시코돈과 가까울수록 근동질계통내에서 발견되는 다형성의 분포와 비교적 정확한 분리패턴을 보이는 것을 발견하였다. 또한 몇 개의 특정형질과는 고도의 유의성을 보이는 유전자 특이적 분자마커를 발견하였다.

특정형질과 관련된 유전자 특이적 분자마커를 개발하는 과정은 육종의 효율을 높이는 매우 중요한 과정이나 기술적으로 근동질계통간 유전체 수준에서 다형성을 발굴하는 것은 다양한 데이터베이스 마이닝기법을 사용한다 하더라도 결과 예측이 불확실한 작업이다. 더욱이 근동질계통간 다형성이 발견되었다 하더라도 그것이 어떤 형질과 연관되어 있는지를 밝히는 것은 전혀 별개의 실험적 분석이 필요하다. 본 실험에서는 유전체수준에서 높은 유사성을 가지고 있는 자포니카 타입 벼들간의 교배조합을 대상으로 하여 다양한 형질이 분리하는 200여 계통을 대상으로 하여 전사조절유전자집단에서 발굴한 근동질계통간 유전자 특이적 분자마커를 이용하여 46개의 표현형과의 연관관계를 통계 분석하였다. 두 개의 서로 다른 벼 염기서열 데이터베이스가 사용되었으며 다양한 벼 염기서열 데이터마이닝 기법이 사용되었다. 본 실험을 통하여 특정 유전자와 밀접하게 연관된 부위의 다형성은 유전자 개시코돈과 가까울수록 근동질계통내에서 발견되는 다형성의 분포와 비교적 정확한 분리패턴을 보이는 것을 발견하였다. 또한 몇 개의 특정형질과는 고도의 유의성을 보이는 유전자 특이적 분자마커를 발견하였다.

콩의 대표적인2차대사 산물인 아이소플라본은 발아기의 뿌리 및 배축, 종실발달기의 협, 종실생육후기의 종자 등에서 만들어지며 뿌리혹박테리아의 고정, 환경적 요인에 의한 스트레스, 내병충성등과 관련이 있다고 알려져 있다. 콩 종실내 아이소플라본의 함량 및 생합성과정은 양적특성을 보이며 몇 개의 QTL은 보고되어있는 실정이다 .특정형질과 관련된 유전자 특이적 분자마커를 개발하는 과정은 육종의 효율을 높이는 매우 중요한 과정이나 기술적으로 근동질계통간 유전체 수준에서 다형성을 발굴하는 것은 매우 어려운 작업이다. 본 실험에서는 아이소플라본 생합성관련 유전자 특이적 분자마커를 개발하기 위하여 이미 알려진 특정 유전자외 전사조절유전자 집단을 이용하여 콩 종실내 다양한 아이소플라본 함량을 보이는 콩 품종간 다형성을 발굴하였다. 유전자염기서열의 발굴은 두과작물의 모델식물체인 Medicago truncatula 의 데이터베이스를 마이닝하였으며 96개의 국내외 대표적 콩 품종들이 사용되었다. 본 실험을 통하여 특정유전자집단과 다양한 아이소플라본 함량을 가진 콩 품종등간의 다양성이 비교되었다.

To facilitate the introgression of F. esculentum into the traits of F. homotropicum, several accessions of the hybrids between these two species were pollinated with F. esculentum as the recurrent parent. In vitro embryo rescue was performed to increase the recovery of backcross progenies. The F2 generation was more amenable than F1 hybrids to produce backcross progenies. The F1 hybrids were backcrossed twice with common buckwheat (pin-type F. esculentum, recurrent backcrossing). Also, alternate backcrosses with common buckwheat and F. homotropicum (congruity backcrossing) were carried out. Pollen tube growth of BCF1 x F. esculentum (thrum) and F. homotropicum x BCF1 was disturbed penetration exceeded for all initial interspecific hybrids, and its requirement was proportionally lower when the common buckwheat used as the recurrent parent and as the last parent of congruity hybrids. Effects of both common buckwheat and F. homotropicum on seed success rate for hybridization were observed. Growth of hybrid embryos before rescue, regeneration of mature hybrids all increased recurrent and congruity backcrosses, inter-crosses between F1 plants and selected fertile plants of the second congruity backcrosses.

1993년 GATT-UR 협상이 타결됨에 따라 관세화에 의한 농식품 시장의 전면 개방이 불가피하게 되었으며, 2004년 한-칠레와의 FTA 발효를 시작으로 우리 농업은 본격적인 글로벌 개방시대를 맞이하게 되었다. 하지만 콩의 경우 시장개방으로 경쟁력이 위협받아 콩 생산량이 감소추세에 있으며, 매년150만여 톤 이상, 즉 수요량의 90%이상을 수입에 의존하고 있는 실정이다. 본 연구의 목적은 한국에서 재배 가능한 콩 30품종(장수, 신팔달1호, 신팔달2호, 큰올, 만리, 보광, 검정콩1호, 진품1호, 진품2호, 황금, 단백, 광안, 삼남, 은하, 푸른콩, 소명, 소원, 다원, amsoy, 장단백목, jack1, jack2, pecking, 대원, 개척1호, 개척2호, s015, s09, 대풍, 태광)을 대상으로 경기 남부지역인 여주에서 계통별 재배실험을 실시하여 그 생육단계 조사를 통해 경기지역 재배환경에 적응력이 우수한 품종을 알아보는데 있다. 파종일은 2008년 6월 16일이고 계통별 1~2열로 재식밀도는 80cm x 20cm으로 1주 2본을 점파하였다. 현재까지 약 4개월간 재배중이고 재배기간인 2008년 6월 중순에서 10월 중순까지 평균기온, 최고기온, 최저기온, 일사량 및 강수량을 조사했다.생육단계는8월 7일, 10월 1일 총 2회에 걸쳐 조사하였으며, 또한 10월 1일에 품종별 5협(莢)을 취해 협(莢)크기와 입(粒)중 및 입(粒)크기를 측정해 다음과 같은 결과를 얻었다.

Seed color is an important trait affecting flour yield and quality in wheat. Seed color also is either tightly linked to or pleiotropically controls seed dormancy in wheat, because most of the red-seeded wheats are tolerant to pre-harvest sprouting in comparison to white-seeded wheats. Recently, metabolomics approaches have recently been used to assess the natural variance in metabolite content between individual plants, an approach with great potential for the improvement of the compositional quality of crops. Basically, in the study here, the simultaneous proteomic and metablomic approaches are being investigated to identify the expressed proteins of genes and specific metabolism responsible for the expression of red and white colors of seed. Red seed “Jinpum” and white seed “Kumkang” cultivars were used in this study to identify the storage proteins use of 2-DE, MALDI-TOF/MS. Here we optimize tissue extraction methods compatible with high-throughput, reproducible nuclear magnetic resonance (NMR) spectroscopy based metabolomics. It appears that the proteins expressed were different each other according to two different cultivars from the seeds of hexaploid wheat. Some selected protein spots were identified as follows: B3-hordein, Gamma-hordein-3,bifunctionalalpha amylase/subtilisin Inhibitor. To monitor metabolic profile, wheat grain was ground in liquid nitrogen, ensuring a homogeneous mix of the tissue, solution samples extracted from seed grains of two wheat cultivars were conducted to measurement of metabolite using 1H-1D NMR method. Representative 1H-1D NMR spectra showing the metabolic fingerprints of wheat grain extracted and presented in Fig. The different peaks, observed at 3.4 and 4.3 ppm, were detected and difference in each two cultivars. The metabolic fingerprint of each two wheat cultivars by 1H-1D NMR were analysed using partial least squares (PLS) in mutivariate analysis to confirm metabolic profiling between different cultivars and to screen chemical shift spectrum corresponding to metabolite specifically abundant in each cultivars. Profiling using 1H-1D NMR was applied to measure of abundance of major metabolite. In total metabolites were compared between “Jinpum” and “Kumkang” cultivars. Therefore, NMR based on the metabolic-phenotyping should be mostly applicable to systematic exploration of plant genetic resources as well as to metabolite based on the breeding program involved in crops productivity.

Buckwheat is one of the traditional crops and has become a renewed target of interest or a popular crop as a healthy foodstuff, because it is a good source of cereal protein which is rich with essential amino acids. However, what is critical to our health is that buckwheat contains proteins which cause a allergy. Buckwheat allergy resulting from ingestion is caused by the storage proteins in the grain with molecular weights ranging from 15, 22, 35, 39 and 50 kDa proteins of the inner fractions to low, and there were clear differences in the protein compositions between the inner and outer buckwheat flour fractions. A major allergenic protein of buckwheat is Fag e 1 with molecular weight 22 kDa (BW22KD). Buckwheat allergy is an immunoglobulin E (IgE)-mediated hypersensitive response capable of causing anaphylactic shock. Buckwheat seeds were dissected to endosperm and embryo. From each fraction we extracted proteins and analyzed extracts by SDS-PAGE and 2-DE. On electrophoregrams of endosperm proteins, 6 intense bands were detected. The most intense corresponded to molecular weights ranging from 54 to 65 kDa. These proteins have been reported not to be allergenic. We show here that the allergenic buckwheat seed proteins are found only among embryo proteins. No allergenic proteins were found in the buckwheat endosperm. The results presented here lead to the proposal that patients with hypersensitivity to buckwheat flour should use only fine flour from buckwheat endosperm, as this fraction contains no allergenic proteins. At present, specific protein spots will be selected and in-gel digested for MALDI-TOF-TOF/MS analysis.

The purpose of this study is to develop the EEG (Euchromatin Enriched Genomic) DNA library of wheat, barley, rye and oat. Mcr A and Mcr BC system in DH5 alpha bacteria cell line and Kuemkangmil, Olbori, Olhomil and Olgwiri were used for materials in our experiments. EEG colonies have been constructed by using junk DNA exclusion. We analyzed the genetic information of the colonies using blast searches of NCBI and GRAMENE web sites. One hundred eighty-four, 65, 79 and 119 STS primer pairs were developed using sequencing data of selected colonies in Kuemkangmil, Olbori, Olhomil and Olgwiri respectively. Twenty-eight and forty-two percent of designed primer pair showed polymorphism using six endoucleases in Kuemkangmil, Olbori, Olhomil and Olgwiri germplasm respectively. These primers could be useful for specific allele tagging in mapping populations and germplasm and for the study of functional genomics of wheat, barley, rye and oat.

본 실험은 SSR marker를 이용하여 밀 자원의 근연관계 및 유전자연관지도 작성의 기초 자료가 되는 anchor marker를 탐색하고자 수행되었다. 실험재료는 금강밀 등 41개 국내육성 품종 및 계통과 Genaro81 등 외국품종 7개 등 총 48개 자원을 사용하였고, SSR marker는 밀의 21개 염색체에서 각각의 short arm에 존재하는 3개와 long arm에 존재하는 5개 SSR marker를 선발하여 총 168개의 SSR primer set을 구성하였다. 이 중 108개의 SSR marker에 대한 PCR 실험결과 92개에서 다형성을 보였으며, 국내외 밀 자원은 SSR marker에 따라 1~7개의 loci에서 유전자다형성을 보였다. 선발된 SSR marker들은 국내외 밀 자원의 분류 및 mapping 집단의 anchor marker로서 이용될 수 있을 것이다.