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"protein"

Research Articles

Early selection of grain quality traits in rice (Oryza sativa L.), is essential to improve the yield and quality of this staple crop. We analyzed four key traits—protein content, grain filling rate, height, and panicle length—in 85 Korean cultivars. Through whole- genome resequencing we identified 12,718,879 raw single nucleotide polymorphisms (SNPs); after PLINK-based quality control (bi-allelic selection, call rate≥0.90, MAF≥0.03), ~2.20 million high-quality SNPs remained for machine-learning (ML) pre-screening. To rank the features (without marker-level inference), we applied a liberal univariate PLINK case-control scan using the top and bottom 30% per trait. We also analyzed associations with a linear mixed model (GCTA v1.93.2, MLMA; fixed covariates: ecotype, PC1, PC2; random effect: GRM) to verify calibration under population structure; with n=85, no genome-wide significant hits were detected, and QQ-plots indicated adequate calibration (per-trait effective tests m≈1.54-1.57 million under stricter filters). The random forest feature importance prioritized 26, 51, 19, and 20 core SNPs for the four traits, respectively. Across the algorithms, the best models achieved mean accuracies of 81.8% (protein content), 81.0% (grain filling rate), 73.1% (height), and 94.0% (panicle length). All selected SNPs met the Fluidigm array design requirements, supporting its deployment as a compact, genotype-based panel for early selection and a practical step toward digital breeding in rice.

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Comparative Analysis of Chloroplast Promoters for Optimized Production of Recombinant Proteins in Plants
식물에서 재조합 단백질의 최적 생산을 위한 엽록체 프로모터의 비교 분석
Juho Lee, Woo-Hyun Jeong, Seon-Kyeong Lee, Kyeong-Ryeol Lee, Sichul Lee, Beom-Gi Kim
Korean. J. Breed. Sci. 2025;57(4):359-372.
Published online December 1, 2025
DOI: https://doi.org/10.9787/KJBS.2025.57.4.359

Plant-based production of recombinant proteins has emerged as an efficient and cost-effective alternative to microbial fermentation and mammalian cell culture systems. Chloroplasts harbor high plasmid copy numbers and can be stably transformed, making them efficient platforms for protein production. In the present study, we used green fluorescent protein (GFP) as a reporter to compare the three major chloroplast promoters (rrn, psbA, and rbcL) involved in protein production in Nicotiana tabacum cv. “Petit Havana.” Three chloroplast transformation vectors were constructed, each regulated by a different promoter, and the transformation was performed via biolistic particle bombardment. Transformants were selected based on spectinomycin resistance and were confirmed by PCR. Among the three promoters, psbA showed the highest transformation efficiency and protein expression levels. Reverse transcription quantitative PCR showed that the mRNA levels (relative to Actin) for psbA (218.21±19.64) were nearly twice that of rbcL (126.60±8.78), and five times that of rrn (43.27±1.57). This transcriptional hierarchy was also observed at protein level. Immunoblotting showed the GFP levels (relative to psbA) were: psbA (1.00), rbcL (0.87), and rrn (0.77), whereas quantification through ELISA revealed relative GFP concentrations of: 616.2±28.7 ng/g LFW for psbA, 510.3±32.4 ng/g LFW for rbcL, and 338.9±100.2 ng/g ng/g LFW for rrn. These quantitative results demonstrate the importance of promoter selection for efficient expression of recombinant proteins in chloroplasts and show that the psbA promoter is suitable for high-efficiency chloroplast expression systems, providing a foundation for advancing plant-based molecular farming.

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Articles

밀의 면역원성을 줄이기 위한 최신 연구 동향 소개
Latest Research Trends in Reducing the Immunogenicity of Wheat
Sora Lee, Jong-Yeol Lee, Sichul Lee, Jae-Ryeong Sim, Sewon Kim, Woo-Suk Cho
Korean. J. Breed. Sci. 2025;57(2):65-89.
Published online June 1, 2025
DOI: https://doi.org/10.9787/KJBS.2025.57.2.65

Wheat is a fundamental staple crop worldwide, contributing significantly to global food security due to its versatility and nutritional value. However, gluten proteins, including gliadins and glutenins, have been implicated in various health problems, such as celiac disease, non-celiac gluten sensitivity, and wheat allergies. These disorders affect a wide variety of people globally, creating demand for wheat varieties that balance high-end-use quality with reduced immunogenic potential. This review examines the molecular and genetic mechanisms that regulate gluten protein synthesis, highlighting recent advances in genomic and mutagenic approaches aimed at modifying gluten proteins to enhance the health and quality traits of wheat. Technologies such as RNAi and CRISPR/Cas9 offer promising avenues for reducing wheat immunogenicity without compromising its functional properties in food production. This study also examines the challenges and prospects of utilizing these genetic tools to develop wheat varieties that achieve the dual
objective
s of enhanced health outcomes and high product quality.

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듀럼밀 유전자원의 질소시비 수준에 따른 농업형질 변이 분석
Variations in Major Agronomic Traits of Durum Wheat Germplasm Under Different Nitrogen Fertilization Levels
Hosun Cheon, Sun-Hwa Kwak, Sieun Choi, Sukyeung Lee, Jinhee Park, Kyung-Min Kim, Chul Soo Park, Youngjun Mo
Korean. J. Breed. Sci. 2024;56(3):281-291.
Published online September 1, 2024
DOI: https://doi.org/10.9787/KJBS.2024.56.3.281

Breeding wheat cultivars with high nitrogen use efficiency is crucial for sustainable agriculture. In this study, 130 durum wheat accessions from 53 countries were cultivated under two nitrogen fertilization levels to investigate variations in agronomic traits and identify accessions suitable for low-nitrogen conditions. Under no nitrogen fertilization, SPAD value (-99.2), flag leaf length (−15.8 mm), grain area (−0.52 mm2), grain length (−0.27 mm), thousand-grain weight (+5.2 g), and grain protein content (−4.2%) showed significant differences compared to that under standard nitrogen fertilization. Among these traits, the SPAD value exhibited a strong positive correlation with protein content in the absence of nitrogen fertilization, suggesting its potential as an indicator for selecting germplasm with superior nitrogen use efficiency. Hierarchical cluster analysis, based on the differences in the six traits under the two nitrogen fertilization levels, classified the 130 durum wheat accessions into three groups (Groups 1, 2, and 3). Accessions in Group 2 exhibited superior adaptability under no nitrogen fertilization compared with those in the other two groups. We identified seven accessions in Group 2 that exhibited minimal decreases in SPAD values and protein content under no nitrogen fertilization for use in wheat breeding programs aimed at developing cultivars with high nitrogen use efficiencies.

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New Cultivar Developed

단백질 함량 증가 고품질 특수미 ‘드리미5호’ 육성
Breeding of a Rice Cultivar “Drimi5ho” with High Protein Content and High-Quality Specialty
Jae-Ryoung Park, Eun-Gyeong Kim, Yoon-Hee Jang, Gang-Seob Lee, Kyung-Min Kim
Korean. J. Breed. Sci. 2022;54(4):421-432.
Published online December 1, 2022
DOI: https://doi.org/10.9787/KJBS.2022.54.4.421

Rice is a staple food in most countries, and thus, diversifying its value is necessary. Recently, the focus on good health has increased, and rice cultivars with improved properties should be bred based on consumer demands. At the Plant Molecular Breeding Laboratory of Kyungpook National University, “Hwayeong” was tissue-cultured to derive mutant populations. In the mutant lines, “P35” with high protein content and excellent agricultural traits, was backcrossed with “Ilmi” three times, and the final selected line was called “Drimi5ho.” The heading date of “Drimi5ho” was August 11 (107 d after sowing), which was 5 d later than that of “Ilmi” (August 5, 101 d after sowing). The culm length of “Drimi5ho” was 71 cm, the panicle length was 18 cm, the number of panicles per hill was 13, the number of spikelets per panicle was 113, the ratio of ripened grain was 90.5%, and the 1,000-grain weight was 22.6 g. When "Drimi5ho" was planted ordinary plantation, the yield in the southern mountainous area was 526 kg/10a. Lodging was not observed when “Drimi5ho” was grown in the field with standard fertilization (N-P2O5-K2O = 9-4.5-5.7 kg/10a). This rice cultivar was resistant to leaf/neck blast, bacterial blight (K1, K2, and K3), rice stripe virus, rice black-streaked dwarf virus, and rice dwarf virus. The milled grain of “Drimi5ho” was clear and transparent without any chalkiness; moreover, the grain shape was round and short. Although it is a specialty rice with increased protein content, the taste of the rice and the grain quality were excellent (Registration No. 6829).

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Articles

밀 유전자원의 단백질 특성 분석 및 글루텐 단백질 조성 평가
Analysis of Protein Properties and Gluten Protein Composition Evaluation of Wheat Genetic Resources
Myoung Hui Lee, Changhyun Choi, Kyeong-Hoon Kim, Jae-Han Son, Jinhee Park, Go Eun Lee, Jun Yong Choi, Chon-Sik Kang, Jiyoung Shon, Jong-Min Ko, Kyeong-Min Kim
Korean. J. Breed. Sci. 2022;54(4):245-259.
Published online December 1, 2022
DOI: https://doi.org/10.9787/KJBS.2022.54.4.245

Gluten proteins in wheat grains are generally considered one of the most important factors in determining dough properties and bread quality. In this study, wheat protein quality characteristics were investigated in 607 varieties collected from seven countries grown in a South Korean wheat breeding field for two years. The average protein content was 12.2±1.86%, and the sodium dodecyl sulfate-sediment volume (SDSS) was 46.9±8.39 mL. HI-LINE had the highest protein content (18.3±0.35%) and SDSS (76.7±1.98 mL), while both NE 84557 and Iksan 374 showed small deviations in protein content and SDSS. Protein content and SDSS values were higher in Ax2*+By8 and By9+Dy10 combinations at Glu-A, Glu-B1, and Glu-D1 loci of high molecular weight gluten subunit (HMW-GS) than in other combinations. However, no difference in Glu-A3 and Glu-B3 loci in LMW-GS was observed. Furthermore, in HMW-GS, the composition of Glu-D1 Dy10 and Dy12 had a greater effect on protein quality than Glu-B1 By8 and By9 when the allele of Glu-A1 had Ax2*. Significant differences were found between Dy10 and Dy12 genes of the HMW-GS Glu-D1 and between protein content and SDSS, but not among others. These results suggest that Glu-D1 is extremely important for improving protein quality in HMW-GSs. As a result of this study, HMW-GS allele selection using functional markers, protein content, and SDSS investigation are expected to enable the development of varieties with high protein quality that are stable amid various environmental changes.

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두부 제조용 콩 육종 계통 선발을 위한 두부 간이 제조법 개발
Development of Fast and Simple Processing Method to Evaluate Tofu Traits in Soybean Breeding System
Jiyoung Jung, Ji-Min Kim, Taeklim Lee, Jinho Heo, Ilseob Shin, Ju Seok Lee, Sungtaeg Kang
Korean. J. Breed. Sci. 2022;54(1):34-42.
Published online March 1, 2022
DOI: https://doi.org/10.9787/KJBS.2022.54.1.34

Tofu is produced by mixing condensed soymilk with a coagulant. It has a low calorie and high protein and contains various important vitamins and minerals; therefore, consumption of tofu as a healthy and dietetic food has increased worldwide. For systematic application in breeding programs to increase tofu yield, evaluation of tofu characteristics is essential. To test the various characteristics of tofu, typically, a considerable amount of seeds (approximately 100 g, is required. Here, we propose a simple tofu quality test method requiring a small amount of seeds (15 g), which is 75% less than the amount needed for conventional testing. Tofu was manufactured using 20 soybean varieties with a conventional method (i.e., a tofu making machine, TM) and four simple methods: natural filtering (NF), weight pressure (WP), hand pressure (HP), and rotation machine (RM). The correlation between the five methods was assessed with respect to each tofu characteristic, and the HP and RM methods were statistically similar to the conventional methods, whereas the other methods showed differences. Regarding tofu yield, the HP and RM methods showed significantly higher correlation coefficients of 0.79 and 0.78, respectively, and the coefficient of variation between replications was also low. Therefore, the HP and RM methods were the best to easily characterize the tofu yield using a small amount of seeds. Considering the efficiency of the test, the RM method appears to be more effective than the HP method for testing multiple lines. These results will be used for the identification of quantitative trait locus/genes related to tofu quality and marker-assisted selection of breeding new soybean varieties.

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A statistical analysis of 9,771 non-glutinous rice in breeding line germplasm collected from Korea (2,836), China (2,136), Japan (1,219), and the Philippines (1,213) was conducted using normal distribution, variability index value (VIV), analysis of variation (ANOVA) and Ducan’s multiple range test (DMRT) based on the data obtained from NIRS analysis. According to the normal distribution, the average protein content was 7.9%, and non-glutinous rice ranging over 10% amylose had 23.6% average content. Most resources were between 5.3 and 10.5% in protein content, and 15.7 and 31.5% in amylose content. The VIV was 0.54 for protein, and 0.83 for amylose. The average amylose content was 25.18%, 24.54%, 22.08%, and 21.47% in Filipino, Chinese, Korean, and Japanese resources, respectively, wheereas the average protein content was found to be 8.19%, 7.79%, 7.58%, and 7.42% in Filipino, Chinese, Korean, and Japanese resources, respectively. The ANOVA of amylose and protein content showed significant differences at the level of 0.01. The F-test value was 412.2 for amylose content, and 108.4 for protein when compared with the critical value of 3.78. The DMRT of amylose and protein content showed significant differences (p<0.01) among resources from different countries. The Filipino resources had the highest level of amylose and protein content, whereas; the lowest level of amylose and protein content were found in Japanese when compared with resources of other origins. These results are recommended as helpful materials in the field of breeding.

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This study was carried out to investigate the utilization value of legume crops collected in tropical and subtropical areas. We examined agronomic traits to assess domestic adaptability and evaluated useful components of foreign legumes. We used a total of 201 genetic resources of three legumes, consisting of 68 lentils, 72 chickpeas and 61 guars. The average number of days to flowering of the three legumes ranged from 56.7 to 60.8 days; the shortest in guar and longest in chickpea. The average number of days to growth of the three legumes ranged from the shortest 86.8 days in lentil, to the longest 163.9 days in guar. The maturation period of the three legumes lasted from the end of May until mid-September, based on sowing in March. However, the average yield of lentil was very low, ranging from 0.5 g to 30.6 g, with an average 16.4 g based on 10 plants per accession. The average 100 seed weight of the three legumes was 2.2 g for lentil, 22.9 g for chickpea, and 3.8 g for guar. The crude protein content ranged from 14.1% to 32.4% with an average of 20.4%, the highest for guar and the lowest for chickpea. The average crude oil content in the three legume crops was generally low, ranging from 0.8% in lentil, to 4.3% in chickpea. The average dietary fiber content in the three legume crops varied from 15.7% to 50.7%. Guar was the highest source of fiber, followed by chickpea (19.3%) and lentil (15.7%). From the agricultural traits analysis, chickpea and guar could grow domestically. However, lentil was difficult to flower and fruit normally during the warmer season after May. Therefore, lentil should be considered for late summer cropping during the cool season. The physicochemical properties of the three legumes seem to be useful as they are similar to, or better than, those of the control common bean.

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밀 반수체 계통에서 오메가-5 글리아딘 돌연변이의 선발 및 동정
Screening and Identification of Omega-5 Gliadin Mutants in Wheat Doubled-Haploid Lines
You-Ran Jang, Chon-Sik Kang, Sun-Hyung Lim, Jong-Yeol Lee
Korean. J. Breed. Sci. 2018;50(3):181-192.   Published online September 1, 2018
DOI: https://doi.org/10.9787/KJBS.2018.50.3.181

Gliadin proteins are a major component of gluten proteins and important determinants of bread-making quality by conferring the viscosity and extensibility of dough, but also present significant health problems for consumers with wheat-related diseases like celiac disease or wheat allergies. In order to solve this problem, we conducted RP-HPLC analysis to profile gliadin fractions for screening the mutants deficient in gliadins from 122 wheat doubled-haploid (DH) lines cultivated by the National Institute of Crop Science. Comparing the RP-HPLC chromatogram of 122 DH lines with those of the respective parents, we found that some peaks of omega-5 gliadin were not present in 28 DH lines. Further analysis using SDS-PAGE and A-PAGE showed that the omega-5 gliadin in the parental varieties had two to three bands, but only one band in the absent 28 DH lines. The relative expression levels of all gliadin groups in the parental and mutant lines were also examined by RP-HPLC. Our study contributes to establishing a method for the rapid screening and identification of mutants missing gliadins as major epitopes of wheat-related disease in many wheat genetic resources and breeding lines as valuable information to other researchers.

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Gliadin proteins, which are a component of gluten and confer viscosity and extensibility on wheat dough, are major determinants of wheat processing suitability and also present dietary problems for consumers with celiac disease or wheat allergies. In this study, gliadin proteins of the hexaploid wheat variety ‘Chinese Spring’ (CS) and of its nullisomic-tetrasomic (NT) and ditelosomic (DT) lines missing group 1 and 6 chromosome, were analyzed using LabChip GXII Touch 24 within 1 min per sample. The chromatogram pattern analysis of gliadin proteins from group 1 aneuploid lines (N1AT1B, N1AT1D; N1BT1A, N1BT1D; N1DT1A, N1DT1B) missing 1A, 1B and 1D chromosomes respectively, from CS showed that 24, 25 and 26 sec peaks of CS, presuming to be ω5-, ω1,2- and γ- gliadins, were disappeared. The analysis of group 6 aneuploid lines (N6AT6B, N6AT6D, 6AL; N6BT6A, 6BL; N6DT6B, 6DL) missing 6A, 6AS; 6B, 6BS; 6D, 6DS chromosomes respectively, from CS indicated that 22, 25 and 26 sec peaks of CS, presuming to be α-/β- gliadins, were disappeared. The results of this study will be applicable to high-throughput screening of wheat gliadin mutants among wheat breeding lines and genetic resources for the development of allergy - reduced wheat.

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This study was investigated to develop mass evaluation system for the contents of crude protein, oil and fatty acid in soybean germplasm using NIRS. NIRS equations were created with 345 soybeans, multiple correlation coefficients of crude protein, oil, palmitic, stearic, oleic, linoleic and linolenic acid between data obtained from NIRS and quantitative analysis were 0.983, 0.969, 0.592, 0.514, 0.978, 0.961 and 0.957, respectively. Equation statistics indicated that contents of crude protein, oil and unsaturated fatty acid except palmitic and stearic acid in soybean seed were suitable for determination by NIRS. Those NIRS equations were applied to examine crude protein, oil and unsaturated fatty acid of 854 soybean landraces from Korea. The average contents and ranges of crude protein and oil were 39.2% with a range of 33.7-47.0% and 15.0% with a range of 9.8-20.3%, individually. In addition, those of oleic, linoleic and linolenic acid were 21.4% with a range of 12.1-30.2%, 55.6% with a 47.8-62.3% and 8.1% with a range of 5.9-10.7% respectively. We conducted quantitative analysis to reconfirm with IT154552 (45.1%) and IT023955(46.9%) above 45% of crude protein, the results were similar from NIRS (45.2%, 47.0%). NIRS data for protein from this study made no difference with lab data, which would be useful for mass evaluation. There was negative correlation (-0.203) between crude protein and oil, positive correlation (0.379) between crude oil and oleic acid, and significantly negative correlation (-0.879) between oleic and linoleic acid.

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Evaluation of Gliadin and Storage Protein Activator (Spa) in Korean Wheat Cultivars
Ji-Eun Kim1, Young-Mi Yoon1,2, Seong Woo Cho1, Chon-Sik Kang2, Jong-Yeol Lee3, and Chul Soo Park1*
Korean. J. Breed. Sci. ;48(3):206-216.   Published online September 30, 2016
DOI: https://doi.org/10.9787/KJBS.2016.48.3.206
Genetic variations of γ-/ω-gliadin and Spa (storage protein activator) in 40 Korean wheat cultivars were evaluated to provide genetic information for improving end-use quality in wheat breeding programs. Korean wheat cultivars were classified into 13 patterns at the Gli-1 locus based on the allelic variation using A-PAGE (acidic-polyacrylamide gel electrophoresis). Seven, five, and six alleles were identified at Gli-A1, Gli-B1, and Gli-D1 loci, respectively. Allele-specific PCR markers for γ-gliadin corresponded to specific allele at Gli-1 loci on A-PAGE, which Gli-A1f, Gli-A1h and Gli-A1l alleles corresponded to GliA1.2, Gli-B1h and Gli-B1f alleles corresponded to GliB1.2 and Gli-D1f, Gli-D1m and Gli-D1o alleles corresponded to GliD1.1. DNA markers for γ-45 and γ-42 also corresponded to the γ-gliadin patterns around 40kDa on A-PAGE, except in Sukang, Ol and Joongmo2003. However, allelic specific PCR markers for ω5-gliadin did not correspond to that of A-PAGE. Three alleles were identified at Spa-A1 locus, whereas there was no variation at Spa-B1 and Spa-D1 loci.
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New Cultivar Developed


Soybean Cultivar for Tofu, ‘Saedanbaek’ with Disease Resistance, and High Protein Content
Hyun Tae Kim1, Jong Min Ko1, In Youl Baek1, Meong Ki Jeon1, Won Young Han1, Keum Yong Park1, Byong Won Lee1, Young Hoon Lee1, Chan Sik Jung1, Ki Won Oh1, Tae Joung Ha2, Jung Kyung Moon1, Hong Tae Yun1, Jong Hyeong Lee3, Jae Keun Choi4, Jae Hyun Jung5, Seung Su Lee6, Yeong Jik Jang7, Chang Ki Son8, and Dal Soon Kang9
Korean. J. Breed. Sci. ;46(3):295-301.   Published online September 30, 2014
DOI: https://doi.org/10.9787/KJBS.2014.46.3.295
A soybean cultivar for tofu, ‘Saedanbaek’, was developed from the cross between ‘MD87L’ with high protein content and ‘SS99237 (Pokwangkong/Suwon193)’ with good seed performance and high yield by National Institute of Crop Science (NICS) in 2010. ‘SS99237-45-2-3-1’ was selected by pedigree selection and designated as the name of ‘Milyang 192’. It was prominent and had good result from regional adaptation yield trials (RYT) for three years from 2008 to 2010 and released as the name of ‘Saedanbaek’. It has a determinate growth habit, white flower, grey pubescence, yellow seed coat, spherical seed shape and medium seed (20.7 grams per 100 seeds). ‘Saedanbaek’ has high content in protein (48.2%), and shows excellent result for making tofu. ‘Saedanbaek’ was resistant to bacterial pustule and moderately resistant to soybean mosaic virus, the major soybean diseases in Korea. The average yield of ‘Saedanbaek’ was 2.28 ton per hectare in the regional yield trials carried out for three years from 2008 to 2010.
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Article

Screening of Soybean Germplasm with High Starch Content
Sung Cheol Koo, Myeong Gi Jeon, Young Hoon Lee, Hyun-Young Kim, Beom Kyu Kang, Jong Min Go,In Youl Baek, Hong Tai Yun, In Youl Baek, and Man Soo Choi*
Korean. J. Breed. Sci. ;46(1):52-57.   Published online March 31, 2014
DOI: https://doi.org/10.9787/KJBS.2014.46.1.052
Starch is the most important carbohydrate in the human and animal diets and it also has numerous industrial applications. Starch is widely used as a key material for processed foods such as bread, pancakes, cereals, noodles, pasta, porridge and tortilla. Increasing the starch content in food crops is one of the important targets for crop breeding. We used a starch-iodine test to identify varieties with high starch content in 2354 soybean germplasm collection and found a total of 126 germplasm that showed a strongly positive reaction. We quantified starch content from highly stained 7 soybean accessions by standard method (glucose-oxidase). As results, starch contents of 7 soybean germplasm were 2.81~4.55%, whereas the weakly stained controls showed low starch content with less than 1%. We also measured the contents of protein, fatty acid, water potential and free sugar in 7 soybean germplasm with high starch content. High starch germplasm showed lower protein content, while low starch germplasm showed higher protein content. Our results suggest that the increasing of starch content could affect protein level in soybean seed.
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