The aim of this study was to select the abiotic tolerant sorghum mutants using chlorophyll a transient OJIP analysis of PSⅠ and PSⅡ so called Kautsky’s effect within 1 second. It was clearly identified that wwt-and drought tolerant sorghum mutants could be classified by wet factor index(WFI). On the basis of WFI, wet tolerant sorghum matants were classified as follows; Ⅰ group, MUT534 bmr/new, MUT525 bmr; Ⅱ group, M2P1207 bmr, 25M2-0404 bmr, MUT371 bmr24, unknown bmr22, 10M2-0775 bmr, MUT135 bmr23; Ⅲ group, M2P0411 bmr, MUT641 bmr, M2P1064 bmr36, MUT855 bmr, 25M2-0137 bmr/new, MUT436 bmr, M2P0929 bmr, 25M2-0026 bmr, 10M2-0387 bmr, 25M2-0173 bmr/new; Ⅳ group, 25M2-0698 bmr. In conclusion, for the selection of wet tolerance, four photochemical parameters such as Electron transport flux until PSI acceptors per PSII(RE1o/RC), Performance index for energy conservation from photons absorbed by PSII antenna, until the reduction of PSI acceptors(PI_total ABS), Driving force on absorption basis(DF_total ABS) and Electron transport flux from QA to QB per PSII(ETo/RC) were important photochemical parameters deduced from maximum quantum yield and electron transport efficiency.

SELDI-TOF MS를 활용한 저분자 펩타이드 분석을 위해 서는 분석시료에 대한 최적화 분석조건을 확립하는 것이 필수적으로 선행되어야 한다. 본 연구는 수수 종자 내 존재하는 10 kDa 이하의 저분자 펩타이드를 프로파일링하기 위하여 활용된 SELDI-TOF MS의 최적화 분석조건을 확립하는데 있다. 분석조건은 (1) 프로테인 칩: CM10(weak cationexchanger), Q10(strong anion exchanger), (2) 바인딩 버퍼의 희석배수: 1/2, 1/5, 1/10, 1/20, 1/50, 1/100, 1/200, (3)Q10의 바인딩 버퍼 강도: 10 mM, 100 mM, (4) 단백질 추출버퍼: sodium borate, sodium borate + acetone, phenol,TCA 버퍼로 하였다.1. 바인딩 버퍼의 희석배수는 CM10과 Q10 모두 1/20과1/50이 최적화로 나타났다.2. Q10의 바인딩 버퍼 강도는 농도가 약한 10 mM에서 더많은 피크가 검출되었다.3. SELDI-TOF MS 분석에 적합한 수수 종자단백질 추출버퍼로는 2~10 kDa 범위에서는 sodium borate 버퍼와10~20 kDa 범위에서는 phenol 버퍼로 분석되었다.

Cereal seeds, sorghum, foxtail millet, hog millet, adlay, and corn are traditionally used as health assistant as well as energy supplying food in Korea. While beneficial phytochemicals to human have revealed in cereals, the information on peptides from cereals is far less accumulated than major reserve protein. Here, we analyzed peptide profiles using surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF MS) in cereal seeds for construction of peptide information and attempted to develop peptide biomarkers for cereal identification. To optimize the analysis condition of SELDI-TOF MS, the effect of dilution factor on binding affinity to protein chips was tested using CM10 and Q10 arrays. Peptide clusters were significantly different at the level of 0.01 p-value. Peak spectra were the most stable in 1:50 of dilution factor in both chip arrays. Numbers of detected peak of 5 cereal seeds were 131 in CM10 and 74 in Q10 array. Each cereal was grouped as a cluster and well discriminated into different cluster in the level of 0.01 p-value. Numbers of potentially identified peptide biomarkers are 11, 13, 9, 5 and 12 in sorghum, foxtail millet, hog millet, adlay and corn, respectively. This study demonstrates that each cereal seed have own distinguishable specific peptides although their function are not identified yet in this study. In addition, the proteomic profiling using SELDI-TOF MS techniques could be a useful and powerful tool to discover peptide biomarker for discrimination and assess crop species, especially under 20 kDa.

Discovery, identification, and informatics of low molecular weight peptide are extensively rising in the field of proteomics research. In this study, we analyzed protein profiles to discover peptide based biomarker for twelve different soybean seeds with three different agronomic types using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS). For optimization of SELDI-TOF MS in soybean seed proteome analysis, four different extraction buffers were tested with urea solubilization buffer, thiourea/urea solubilization buffer, phenol extraction buffer, and modified trichloroacetic acid (TCA)/acetone precipitation/urea solubilization extraction buffer. Two different type of ProteinChip arrays, cation exchange (CM10) and anion exchange (Q10), applied to profile peptides. Among the four different extraction buffers, phenol extraction was selected to protein extraction methodology. Numbers of detected peak cluster in twelve soybean seeds were 125 at CM10 and 90 at Q10 array in the mass range from 2 to 40 kDa. Among them, 82 peak clusters at CM10 and 33 peak clusters at Q10 array showed significantly different peak clusters at p<0.00004 (CM10) and p<0.00005 (Q10) among twelve different soybean cultivars. Moreover, 29 peak clusters at CM10 and 17 peak clusters at Q10 array were detected in all cultivars as an ‘universally existed peptide’. In comparison with three different agronomic types, total of 55 peak clusters (CM10) and 23 peak clusters (Q10) were significantly different peak clusters at p<0.00004 and p<0.0001, respectively. In these probability levels, soybean seeds were well discriminated into different cultivar and different type with each other. Also we could find several specific peptide biomarkers for agronomic type.

Development of transgenic plant with desirable traits to cultivated plant is one of the important procedures in plant molecular breeding. However, applicable assessment of transgenic plant in laboratorial scale is not much except cultivating transgenic plant for a whole life in field condition. Here, we analyzed chlorophyll fluorescence in three transgenic soybean lines with AtMYB44 transcription factor for assessment of photosynthetic activity under abiotic stresses such as drought. Soybean varieties used in this study were ‘Bert’ and ‘Bert’ derived three transgenic soybeans, ‘AtMYB44 CM35101’, ‘AtMYB44 CM2471’, and ‘AtMYB44 CM4481’. Analyzed five different chlorophyll fluorescence variables are maximum PSII quantum yield (QY_max), steady state PSII quantum yield (QY_Lss), steady state non-photochemical quenching (NPQ_Lss), coefficient of photochemical quenching in steady-state (Qp_Lss), and fluorescence declineratio in steady-state (Rfd_Lss). To determine main chlorophyll fluorescence variable affected by abiotic stress, principal component analysis (PCA) was conducted with five chlorophyll fluorescence variables measured from four varieties. QY_Lss and NPQ_Lss were main chlorophyll fluorescence variables to evaluate abiotic stress, particularly in drought stress. In comparison with transgenic soybean lines based on chlorophyll fluorescence variables, ‘AtMYB44 CM2471’ and ‘AtMYB44 CM4481’ are more tolerant to drought than the others. Interestingly, three transgenic soybean lines which have a same AtMYB44 gene with different regions of chromosome revealed the quite different responses of chlorophyll fluorescence to drought treatment.

To investigate the application of biochemical markers' and small-sample methods using whole-wheat flours for screening in early generation in Korean wheat breeding system, 74 Korean wheats, including cultivars, local breeding lines and experimental lines, were analyzed. Seed storage protein and amylose contents of grains were evaluated. Biochemical makers, including granule bound starch synthase (GBSS), high molecular weigh glutenin subunits (HMW-GS) and friabilin were also evaluated by using one-dimensional sodium dodecyl sulfate-polyacryla-mide gel electrophoresis with a single kernel. The small­sample methods, including modified SDS-sedimentation test (MST), micro-alkaline water retention capacity (AWRC) and whole-wheat flour swelling volume (WSV) were also tested in this study. Protein content, MST and AWRC was 11.0 - 15.8~% , 2.7 - 26.2 ml and 71.9 - 109.7~% , respectively. Apparent and total amylose content and WSV was 20.6 - 25.0~% , 26.1 - 32.4~% and 9.0 - 16.9 ml, respectively. There were highly significant correlations between MST and AWRC (r=0.592, P<0.001), but Korean wheats showed no significant difference in protein content, amylose content and small-sample methods. In the biochemical markers, Korean wheats contained all three GBSS encoded by Wx loci, except for Suwon 252. Korean wheats showed the high frequency (58.1~% ) of 1Dx2.2 + 1Dy12 subunits of HMW-GS. Friabilin band was present in 46 lines (62.2~% ) and absent in 28 lines (37.8~% ). Friabilin-absence lines showed the higher MST (14.9 ml) and AWRC (92.1~% ) value than friabilin-presence lines (8.5 ml and 82.4~% , respectively).

Rice (Oryza sativa L.) plants were cultivated to examine changes in antioxidative defence mechanism induced by elevated ozone levels. Catalase activities in tolerant Jinpumbyeo and susceptible Chucheongbyeo under ozone fumigation were reduced at 5 hrs and 3 hrs after ozone fumigation, respectively. With the increased ozone supply, peroxidase activity in Jinpumbyeo was steadily enhanced whereas in Chucheongbyeo it was not changed. Four SOD-isozymes were detected by NBT staining of native-PAGE. Two isozymes of them were obviously induced by ozone supply, particularly in Jinpumbyeo. The continuous ozone fumigation increased remarkably putrescine levels in leaves whereas it did not affect the levels of spermidine and spermine. In this study, it was implied that ozone in cell inhibits strongly diamine oxidase and thus promotes ethylene biosynthesis which will cause the senescence in rice plants.

We have developed and tested a new method for nondestructive estimation of chlorophyll- and nitrogen-contents in rye leaf. It was found that the relation-ships among nitrogen, chlorophyll content and fresh weight were significantly positive correlated. Nitrogen and chlorophyll content were positively correlated whereas correlation coefficients among R, G, R-B and G-B on the basis of photo-numerical values were negative. We have found that R/(R-B) obtained from data of digital camera is the best criterion to estimate the chlorophyll content of leaves. The regression curves of the relation between R/(R-B) and chlorophyll content were also calculated from the data collected on cloudy days. The coefficients of determination (~textrmr2 ) were ranged from 0.33 to 0.99. In this study, the accuracy in estimating chlorophyll content from the color data of digital camera image could be improved by correcting with R, G, and B values. It is suggested that, for practical purposes, the image values estimated with sufficient accuracy using a portable digital camera can be applied for determining chlorophyll content and nitrogen status in plant leaves.

To observe and analysis ultra-microscopically barley aleurone cell surface, atomic force microscope (AFM) was used. Seed coat of early maturing germplasm, eam9, was dehulled and scanned by non-contact mode. We have obtained the high resolution topographic 3-dimensional image of barley aleurone layer with high resolution. These images showed the membrane proteins in barley aleurone cell. One channel protein and numerous peripheral or integral proteins were detected in a area of 100 ~mu~textrmm2 . Furthermore, we found that their widths were ranged from 50 to 750nm and lengths from 0 to 66 ~mu~textrmm . The thickness of aleurone layer was measured by scanning electron microscope. The thickness at early developmental stage was about 16 and then the aleurone cell enlarged upto 57 ~mu~textrmm ~mu~textrmm at least until 42 days after anthesis. In this study, we firstly reported on the ultrastructural AFM analysis of living aleurone cell as a biological specimen. It was clearly suggested that AFM will become an powerful tool for probing both the structural properties of biological samples