본 리뷰의 목적은 벼 종자 저장단백질 구조분석 및 발현특성분석 결과 종합화를 통하여 종자형질 개선 등의 실용화연구를 위한 기반구축을 모색하는데 있다.
최근 벼 염색체염기서열완전해독 연구 결과를 이용한 유용형질 유전자 분리 및 실용화 연구가 많이 진행되고 있다. 특히 벼 종자 저장단백질은 인류에게는 주요 영양원으로 사용되어지며 종자 발아시에는 식물체 성장을 위한 질소원으로 사용되어진다. 벼 종자 저장단백질의 분류는 용매에서의 용해도에 따라 약산성 및 알카리 용해성의 glutelin, 알코올 용해성의 prolamin, 염 용해성의 globulin으로 나눈다. 벼 염색체 상에는 11개의 glutelin 유전자와 33개의 prolamin 유전자가 존재하며 prolamin 유전자의 경우 5번 염색체 15 Mb 부위에 15개의 유전자가 위치하였다. 이와 같이 종자저장단백질 유전자들이 동일 염색체 부위에 위치하고 있는 것은 진화학적으로 동일 염색체에서 유래하였거나 유사한 유전자발현 조절영역을 가지고 있음을 의미한다. Globulin 유전자는 5번 염색체에 단일 유전자로 존재하였다.
마이크로어레이를 이용한 종자저장 단백질 관련 유전자의 조직 특이 발현 양상을 분석한 결과 glutelin과 대다수의 prolamin 합성 유전자는 종자배유에서만 발현을 하였으며 소수의 prolamin과 globulin 합성 유전자는 종자배유와 발아종자에서도 발현을 나타내었다. 종자 저장단백질의 프로모터부위를 분리한 후 종자에서의 발현 양상을 분석한 결과 glutelin type C1 프로모터가 종자의 전체 부위에서 발현을 나타내었으며 glutelin type B5와 α-globulin 프로모터가 많은 양의 발현을 나타내었다. 본 리뷰를 통하여 벼 종자 저장단백질의 구조및 발현특성 연구 진행사항을 살펴보았다. 이러한 연구 동향분석은 종자형질 개선 및 물질생산 등의 실용화 연구를 수행하는 연구자들에게 최근의 연구 현황을 제공할 수 있을 것으로 생각된다.
인간과 가축의 영양을 위한 식물성 단백질의 주요 공급원은 콩이며 콩 단백질은 영양 및 기능성면에서 우수하여 소비가 점차 증가하고 있다. 그러나 콩 단백질에는 알러지를 일으키고 영양가치를 떨어뜨리는 성분도 포함되어져 있다. 7S 및 11S 글로블린은 콩 저장단백질의 대부분을 차지하며 7S는 영양가치가 떨어지고 7S의 함량을 줄어든 콩 계통 육성에 대한 관심이 높아지고 있다. 7S 성분중의 하나인 α′-subunit의 유전양상을 파악하기 위하여 진품콩2호와 PI506876의 교배로부터 98개의 F2 종자가 얻어졌다. SDS-PAGE로 각각의 종자를 분석한 결과 α′-subunit을 가진 종자가 70개였고 결핍된 종자가 28개였다. 이러한 유전양상은 단인자 유전원칙 (χ2 = 0.667, P=0.414)과 일치하여 콩 종자에서 7S의 α′-subunit 단백질은 한 개의 유전자에 의해서 좌우되었다. 이 결과는 7S 단백질 함량이 줄어든 콩 계통 선발에 유용하게 활용될 것으로 기대된다.
본 연구에서는 단백질 함량이 높은 들깨박을 기능성 식품소재로서의 활용가능성을 확인하기 위하여 단백질분해효소 및 한외여과를 이용하여 가수분해물과 펩타이드 분획을 제조하고 이들의 항산화 활성을 측정하였다. 먼저 들깨박단백질의 가수분해물 생산을 위한 최적 효소를 선정하기 위해 단백질분해효소 7종을 이용하여 효소 반응 후 가수분해도를 측정한 결과, flavourzyme이 가장 높은 가수분해율을 나타내었다. Flavourzyme에 의한 들깨박단백질 가수분해물을 얻기 위한 최적 조건은 pH 7.0, 50℃, 효소농도 10 unit, 가수분해시간은 4시간으로 결정되었다. 들깨박단백질 가수분해물을 한외여과 후 얻은 각 분획의 수율은 1 kDa 이하가 45.65%로 유의적으로 가장 높았으며, 그 다음으로5-10 kDa(16.45%), 10 kDa 이상(16.37%), 1-3 kDa(10.86%), 3-5 kDa(10.67%) 순으로 높았다. DPPH 라디칼 소거능은 효소가수분해물이, 환원력은 3-5 kDa 분획물이, superoxide dismutase 유사활성은 1 kDa 이하의 분획물이 가장 높은 활성을 나타내었다. 따라서 들깨박단백질로부터 생산된 가수분해물 및 펩타이드 분획은 각기 다른 항산화 활성 특성을 보여 기능성 식품 소재의 목적에 맞게 선택하여 활용할 수 있을 것으로 판단된다. 또한 향후 식품학적 기능성 평가가 이루어진다면 식품산업에서 다양한 식품재료로의 활용성이 확대될 수 있을 것으로 판단된다.
Soybean [Glycine max (L.) Merr.] is an important source for protein, oil, carbohydrates, isoflavones, and many other nutrients to humans and animals. Lipoxygenase protein is the main antinutritional factor in mature soybean seed. Soybean cultivar, "Choseon" with yellow medium seed size and free of lipoxygenase 2,3 protein was developed. "Choseon" was selected from the population derived from the cross between cultivar, "Jinpum 2" and germplasm, PI408155. "Choseon" has purple flowers, tawny pubescence, a determinate growth habit, and light yellow pods at maturity. The seed of "Choseon" has yellow hilum and yellow seed coat color. Seed protein and oil content on a dry weight basis were 34.2 and 16.9%, respectively. "Choseon" has shown a resistant reaction to soybean necrosis, soybean mosaic virus, Cercospora leaf spot and blight, black root rot, pod and stem blight, and bacterial pustule. "Choseon" matured in 5 October with plant height of 51 cm and a 100-seed weight of 15.1 g. Average yield of "Choseon" was 2.66 (Ton/ha) on the regional yield trials of 2013 at 4 locations. "Choseon" has been registered as a soybean cultivar (registration number: 5985, registration date: April 11, 2016) by Korea Seed & Variety Service, Republic of Korea.
In this study, we analyzed seed storage proteins in order to investigate the main factors related to the eating quality of japonica and tongil-type rice varieties. Sensory evaluation was performed by a trained panel to assess the appearance (color and glossiness), flavor, taste, stickiness, texture, and overall score of nine japonica and three tongil-type rice cultivars. Moreover, the pattern of variation in rice storage proteins was examined by electrophoresis of protein extracts. The electrophoretic pattern of rice proteins showed 16.4 kDa albumin, 26.4 kDa globulin, 34-39 kDa and 21-22 kDa glutelin, and 14.3 kDa prolamin. In terms of storage protein, the varietal differences between japonica and tongil-type rice were found in albumin, globulin, and the α-1, and α-2 sub-units of acidic glutelin. Furthermore, the overall sensory evaluation score was observed to be positively correlated with albumin (0.495 ** ) and globulin (0.567 ** ), and negatively correlated with α-1 glutelin (-0.612 ** ). Therefore, the results indicated that albumin, globulin, and α-1 glutelin can affect the eating quality of japonica and tongil-type rice varieties, with the latter having lower eating quality than the former.
This study was conducted to improve the postharvest storage techniques of managing and storing seeds, totest qualities and viabilities of the seeds and to examine the germination rate and the protein expression of Achyranthesjaponica Nakai. The seeds collected from different areas of Je-Cheon and Gwang-Ju were stored with different temperaturesand durations. Two plant growth regulators and two seed priming were treated to investigate their effect on the germinationrates and the days required for germination. The weight of one hundred seed collected in Gwang-Ju was heavier than thosein Je-Cheon. Seed length collected in Gwang-Ju was also longer about 5.12㎜ than those in Je-Cheon about 4.90㎜ andseed width was longer in Gwang-Ju than those in Je-Cheon. The rates of seed germination in two different collection areaswere higher about 2.9 to 13.0% in Gwang-Ju compared to those in Je-Cheon. Comparing its rates with the storing tempera-tures and durations, they were not clearly different in between 4℃ and 25℃ and they also were gradually decreased withgetting longer storing durations. The germination rates treated by plant growth regulators were higher with GA3 than thosewith Kinetin. The highest seed germination rate was appeared at 50 ppm of GA3. Comparing its rates with different seedpriming, they were relatively higher with KNO3 than those with PEG6000. In protein expression patterns between before thegerminating and after the germinating of seeds, more and clear bands were appeared in the seed after the germination com-pared to those before the germination of seeds, especially 10~20kDa. These results showing more and clear bands weremore clearly appeared in Gwang-Ju compared to Je-Cheon. Comparing the protein expression with plant growth regulatorsand seed primings, GA3 was better expression than those with Kinetin and KNO3 was better than those with PEG6000. Moreand clear bands were closely related to the germination rates of seeds and more detailed studies would be required.
This study was conducted to investigate the quality of seeds, the germination rates and the days required for germination, to examine the patterns of protein expressions during the germination and to improve the techniques of managing and storing seeds and viability of the seeds of Lithospermum erythrorhizon Sieb. et Zucc. After collecting and harvesting seeds, they were classified to white and brown colors of seed coat through testing their seed size, weight, and quality. The germination rates, the days required for germination, and the protein expressions were examined with different colors of seed coats, storing temperatures and durations by treating the different plant growth regulators and primings. One hundred seed weight of white color was heavier about 1.17 g than those of brown one about 0.81 g. The germination rates in white color of seed coat was higher, 3.05 ~ 5.75%, than those in brown one. Its rates were decreased with getting longer in storage durations. There was no big differences on germination rates between storage temperatures. The plant growth regulator of GA3 and Kinetin was affected to improve the seed germination. GA3 increased the seed germination clearly at 25 ppm level, while kinetin increased it gradually from 25 to 100 ppm levels. In germination by seed primings, PEG6000 made higher germination rate with increasing their levels, whereas KNO3 increased the germination until 100 mM level and then decreased it with 200 mM unlike PEG6000. The protein expressed during the seed germination were appeared more and clearer bands in the seed after germination, especially 20 ~ 30 kDa, compared to those in the seed before germination. These results showing more and clearer bands were positively related to the germination rates which were different by seed colors, storage temperatures and durations, and plant growth regulators and primings.
Sorghum seed is traditionally used as health supplements and the secondary food mixed with rice in Korea. While the research of reserve protein in sorghum seed have been carried out in many countries used as major food, much less is known about reserve proteins of Korean local sorghum seeds. To obtain protein characteristics in 20 Korean local sorghum seed, quantitative content of reserve protein was determined after fractionation by modified ‘Osbone’ method and α-kafirin of prolamin was determined by SDSPAGE. Mean albumin, globulin, prolamin and glutelin contents based on total seed protein content of 20 Korean local sorghum seed were 6.2%, 0.9%, 57.9% and 35.1%, respectively. Sorghum cultivar with high prolamin were ‘Whin-susu’, ‘Whin-Chalsusu’, ‘Whanggeum-Chalsusu’, and ‘Daepungshushu’. Sorghum cultivar with high α-kafirin were ‘Whin-susu’, ‘Geumsan-Chalsusu’, ‘Whin-Chalsusu’, and ‘Jangmok-susu’. Among the 20 varieties, ‘Whin-susu’ and ‘Whin-Chalsusu’ were selected as high α -kafirin and prolamin sorghum cultivar, which showed 64.5 and 71.9% of prolamin contents, respectively.
Soybean [Glycine max (L.) Merr.] protein is excellent nutritional factors and is widely used for human and animal feed in the world. Glycinin(11S globulin) and ß-conglycinin(7S globulin) account for storage protein from 70% to 80% in soybean seed. ß-conglycinin is composed of α’, α and ß subunits, encoded by the genes Cgy1, Cgy2, and Cgy3, respectively. β-conglycinin protein exhibits poor nutritional and food processing properties. Lipoxygenase is responsible for the beany flavor and is responsible for the inferior nutritional quality of unheated or incompletely heated soybean meal. The objective of this research was to select F2 seeds with lipoxygenase-free and low content of ß-conglycinin for breeding of high quality soybean cultivar. A total of 582 F2 seeds were obtained from the cross of 10F1(lipoxygenase-free, normal content of 7S protein) and T311(lipoxygenase-present, low content of 7S protein). Lipoxygenase and 7S proteins in mature seeds were detected by SDS-PAGE. The segregation ratio of 3 : 1 for Shr locus (418 Shr_ : 164 shrshr) were observed. This result shows that the Shr locus is controlled by a single gene. Among 164 shriveled seeds, 6 F2 seeds with lipoxygenase-free, α’ subunit-free, and low content of α and ß subunit were selected. Among 418 normal seeds, 6 F2 seeds with lipoxygenase-free, α’ subunit-free, and low content of α and ß subunit were selected. These results will be used in breeding program for improving high quality soybean cultivar.
인체내에서 소화불량 및 알레르기 반응을 일으킴으로써 콩의 품질을 저하시키는 물질인 Kunitz trypsin inhibitor(KTI) 단백질이 결핍되고, P34 단백질을 적게 함유하는 콩 계통을 선발하기 위해서, 현재까지 보고되지 않은 KTI 단백질의 유무와 P34 단백질 함량간의 유전관계에 대한 정보를 얻기 위하여 본 연구에서 얻어진 결과는 다음과 같다. 1. 07B1과 PI567476의 교배를 통해 얻어진 479개의 F2 종자를 대상으로 SDS-PAGE를 이용한 KTI 단백질의 유무를 확인한 결과, KTI 단백질이 존재하는 종자의 수는 353개, 결핍된 종자는 126개로 3 : 1로 분리하였다. 2 Western blot을 이용한 P34 단백질의 함량을 확인 한 결과, P34 단백질 함량이 보통 또는 높은 종자가 363개, 함량이 낮은 종자는 116개로 P34 단백질 함량에 대한 유전분리비는 3 : 1로 나타났다. 3. 전체 F2 종자 479개 중에서 KTI 단백질이 존재하며, P34 단백질 함량이 보통 또는 높은 종자가 266개, KTI 단백질이 존재하고 P34 단백질 함량이 낮은 종자가 88개, KTI 단백질이 결핍이고 P34 단백질함량이 보통 또는 높은 종자가 102개, KTI 단백질이 결핍이며 P34 단백질 함량이 적은 종자가 23개로 9:3:3:1의 분리비에 적합하여 KTI 단백질 유무와 P34 단백질 함량간에는 독립유전을 하였다.
Soybean proteins are widely used for human and animal feeds worldwide. The use of soybean protein has been expanded in the food industry due to their excellent nutritional benefits. But, antinutritional and allergenic factors are present in the raw mature soybean. P34 protein, referred as Gly m Bd 30K, has been identified as a predominant immunodominant allergen. The objective of this research is to identify the genetic mode of P34 protein for the improvement of soybean cultivar with a very low level of P34 protein. Two F2 populations were developed from the cross of "Pungsannamulkong" x PI567476 and "Gaechuck2ho" x PI567476 (very low level of P34 protein). Relative amount of P34 protein was observed by Western blot analysis. The observed data for the progeny of "Pungsannamulkong" and PI567476 were 133 seeds with normal content of P34 protein and 35 seeds with very low level of P34 protein (X2=1.157, P=0.20-0.30). For the progeny of "Gaechuck#1" and PI567476, the observed data were 177 seeds with normal content of P34 protein and 73 seeds with very low level of P34 protein (X2=2.353, P=0.10-0.20). From pooled data, observed data were 310 seeds with normal content of P34 protein and 108 seeds with very low level of P34 protein (X2=0.156, P=0.50-0.70). The segregation ratio (3:1) and the Chi-square value obtained from the two populations suggested that P34 protein in mature soybean seed is controlled by a single major gene. Single gene inheritance of P34 protein was confirmed in 32 F2 derived lines in F3 seeds, which were germinated from the low level of P34 protein obtained from the cross of "Pungsannamulkong" and PI567476. These results may provide valuable information to breed for new soybean line with low level of P34 protein and identification of molecular markers linked to P34 locus.
Kjeldahl method used in many materials from various plant parts to determine protein contents, is laborious and time-consuming and utilizes hazardous chemicals. Near-infrared (NIR) reflectance spectroscopy, a rapid and environmentally benign technique, was investigated as a potential method for the prediction of protein content. Near-infrared reflectance spectra(1100-2400 nm) of coarse cereal grains(n=100 for each germplasm) were obtained using a dispersive spectrometer as both of grain itself and flour ground, and total protein contents determined according to Kjeldahl method. Using multivariate analysis, a modified partial least-squares model was developed for prediction of protein contents. The model had a multiple coefficient of determination of 0.99, 0.99, 0.99, 0.96 and 0.99 for foxtail millet, sorghum, millet, adzuki bean and mung bean germplasm, respectively. The model was tested with independent validation samples (n=10 for each germplasm). All samples were predicted with the coefficient of determination of 0.99, 0.99, 0.99, 0.91 and 0.99 for foxtail millet, sorghum, millet, adzuki bean and mung bean germplasm, respectively. The results indicate that NIR reflectance spectroscopy is an accurate and efficient tool for determining protein content of diverse coarse cereal germplasm for nutrition labeling of nutritional value. On the other hands appropriate condition of cereal material to predict protein using NIR was flour condition of grains.
barley grain and malt is highly related to beer quality, especially hordein is known to be a more significant factor in malting process than albumin. In this study, we proposed selection criteria for high quality malting barley with aid of grain and malt quality parameter scores and storage protein subunit profile informations. Albumin and hordein were extracted and denatured protein subunits were evaluated with malt and grain quality parameters. Total 13 local adaptability test (LAT) lines were planted in four locations (Naju, Iksan, Jeju, and Jinju) and evaluated for malt and beer making qualities. Seventeen germplasms (world collections for high or low seed storage protein content) were also evaluated for biochemical genetic marker. Denatured seed storage protein subunits of albumin and hordein of all tested lines and germplasms were evaluated using 12% 1D SDS-PAGE. Scored data of protein subunit's presence or absence was applied to Agglomerative Hierarchical Clustering (AHC) for statistical analysis. Subunits fractionated within specific molecular weight ranges (97.4-31.0, 66.2-31.0, and 45.0-31.0 kDa) were highly correlated with agricultural characteristics. Several LAT lines showing good performance in agricultural characteristics were clustered in dendrogram constructed by biochemical-genetic assay using XLSTAT. Specific band pattern showed in good performance LAT lines were also observed in some germplasms of world collection having low protein contents which are known to have superior quality in malting. The results would provide selection criteria for high quality malting barley in the malting barley breeding program.
Single seeds of common buckwheat cultivar Suwon No. 1 when subjected to SDS-PAGE revealed very high polymorphism. High variation existed for protein or protein subunits with molecular weight 54-47kDa, 45-25kDa and 16-11kDa. The electrophoregram showed variation for globulin as well as other protein fractions. About 300 proteins were separated by two-dimensional electrophoresis in common buckwheat (Fagopyrum esculentum Moench.) seed. Seed maturation is a dynamic and temporally regulated phase of seed development that determines the composition of storage proteins reserves in mature seeds. Buckwheat seeds from 5, 10, 15, 20, and 25 days after pollination and matured stage were used for the analysis. This led to the establishment of high-resolution proteome reference maps, expression profiles of 48 spots. It was identified 48 proteins from MALDI-TOF/MS analysis of wild buckwheat seed storage proteins. The 48 proteins were found identical or similar to those of proteins reported in buckwheat and other plants; it is belonging to 9 major functional categories including seed storage proteins, stress/defense response, protein synthesis, photosynthesis, allergy proteins, amino acid, enzyme, metabolism, and miscellaneous. It appears that the major allergenic storage protein separated played the important role in buckwheat breeding and biochemical characterization.