Progress 03/15/17 to 03/03/21
Outputs
Target Audience:Soybean breeders, geneticists, soybean growers and associations, marketing firms, plant and food scientists in universities and research laboratories, USDA-ARS soybean researchers, graduate students and the soy food processing industries. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This research provided an opportunity to train one graduate student at the MS degree level at the Mississippi State University. How have the results been disseminated to communities of interest?The research has been presented in three presentations at three separate scientific conferences, including a presentation at the American Chemical Society, and two project director's conferences at the UC-Davis and in Georgia. We anticipate to publish two peer-reviewed journal articles in 2021. In addition, I have presented some findings in the US-Soybean Export Council's Southeastern Asia Regional Soybean Utilization Conferences in Singapore in 2020. I plan to present our research at the University of Missouri's Soybean Conference in 2021. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Seven soybean genotypes from each of three locations (Mississippi, Missouri, and Virginia) harvested in two years were selected to perform protein secondary structure analysis by Circular Dichroism. MS-1, MS-3 and MS-19 with high A3 subunit content; MS-8 and MS-9 with medium level of A3 subunit content; and MS-4 and MS-6 with low A3 subunit content were selected for the analysis. The percentages of α-helix, β-turn, β-sheet and random coil were calculated by DichrWeb software. Each soybean sample solution was prepared in duplicate. Each replicate was scanned 5 times to obtain data for secondary structure analyses. The results showed the percentages of α-helix ranged from 14.28 to 20.05% and the range of β-sheet was from 26.78 to 35.40%, whereas content of β-turn ranged from 16.30 to 24.20%. The sum of β-sheet and β-turn ranged from 47.00 to 57.98%. Random coil ranged from 24.48 to 34.08%. Location and year effect did not show significant variations in α-helix, β-sheet, β-turn, (β-sheet + β-turn) and random coil contents. The results from overall correlation analysis between protein secondary structures and tofu quality showed that the content of β-turn and random coil did not significantly correlate with tofu firmness. The percentage of α-helix negatively correlated with pressed tofu firmness (r = -0.85, probability level less than 0.05). β-sheet exhibited a high correlation with filled tofu hardness (r = 0.76, p < 0.05), but did not show significant correlation with pressed tofu firmness. The total (β-sheet + β-turn) value was highly correlated with both filled and pressed tofu firmness (r = 0.89 and 0.88 respectively, probability less than 0.01). Our results also indicated that both the unfolding of α-helices and the formation of β-sheets during processing may favor the gelation of soy proteins. Thus far, no researchers have characterized the correlationships between the secondary structures (α-helices, β-sheets, β-turns and random coil) in the original soybean seeds and the gel texture of tofu products. Our current study is the first in the literature to show how protein secondary structures may affect tofu gel formation. Cultivation year factor had a significant effect on the percentage of α-helix, β-sheet and β-turn. However, year factor did not have significant influence on random coil and the sum of β-sheet and β-turn. Location and genotype significantly affected all of protein secondary structures. It is important to note that genotype always had more significant effect than location and year. Peptide mapping of soybean proteins in 17 soybean genotypes from three locations over two years was carried out using two-dimensional gel electrophoresis. Proteins in each from three locations in two years was extracted and washed and the native proteins analyzed in the first dimension using isoelectric focusing with pH gradient from 3-10, and the second dimension with SDS-PAGE for molecular mass distribution. Spots represented protein subunits. The spots areas and intensity represented the content of subunits. We selected MS-1 and MS-14 from Missouri in 2018 for peptide map comparison since they have significant differences of 7S and 11S content as analyzed by SDS-PAGE. Aside from differences in content of each subunit in the SDS (molecular size) dimension, the results showed that microheterogeneity existed in peptides of the MS-1 and MS-14 proteins since the shape of some of the major protein subunits along the IEF (isoelectric point) dimension differed and that was due to charge differences of amino acid residues, which caused the movement of peptides molecules towards their respective isoelectric points. It is well known that protein polymorphisms exist in plant seed proteins. However, to what degree protein microheterogeneity affects the overall protein functional property in tofu and other soy foods remained to be investigated. In summary, our successful discoveries of the significant relationships between the composition of glycinin A3 subunit and the secondary structures of proteins in soybeans of various genotypes and tofu quality confirmed our hypothesis and proved the usefulness of using A3 as a protein marker for future genetic improvement of soybean for enhancing soy food quality. This research took four years to plant soybean in three locations and to analyze many physico-chemical properties of many soybean samples. We deeply thank USDA-NIFA-AFRI A1141 Program for providing funding for this research.
Publications
- Type:
Theses/Dissertations
Status:
Under Review
Year Published:
2021
Citation:
Chen, R. 2021. Positive correlation between A3 subunit of glycinin and firmness of tofu made from soybeans grown in three locations over two years. MS Thesis. Mississippi State University. MS State, MS 39762.
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Progress 03/15/19 to 03/14/20
Outputs
Target Audience:The target audience includes soybean growers, marketing firms, tofu processing industry, food industry, soybean food scientists in academia and USDA-ARS scientists, and graduate students. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The study provides the opportunity for the training of one graduate student at the Master degree level. How have the results been disseminated to communities of interest?The results have been disseminated to the scientific communities of interest. It was reported in the NIFA Project Director's conference in the National Plant Breeders Association in Atlanta, Georgia, and in the Agricultural and Food Chemistry Division of the American Chemical Society in 2019. What do you plan to do during the next reporting period to accomplish the goals?We plan to investigate further on how protein secondary structures such as alpha-helix, beta sheet structures, and random coil composition of the soybeans produced in 2017 and 2018 using circular dichroism, and also to study the 2-dimensional electrophoretic patterns of the proteins in soybeans to explore any protein polymorphism in soybeans of high A3 subunit and other glycinin and beta-conglycinin subunits and possible correlations with tofu gel firmness. This would the first in the literature to report how protein subunits and their polymorphism affect gelation of proteins and food quality.
Impacts
What was accomplished under these goals?
Soybean has been traditionally used for making nutritious food like tofu. Producing desirable firmness is important in manufacturing tofu from soybeans. However, which component among the soybean storage proteins is mostly responsible for determining tofu firmness is not fully understood. Our objective is to identify the correlations between seed protein sub-units and the firmness of tofu made from soybeans planted in three locations over two years. Twenty-two soybean Plant introductions from the USDA Soybean Germplasm Collection and eight check varieties were planted in Mississippi, Virginia and Missouri in 2017 and 2018. For each genotype, pressed tofu and filled tofu were made in duplicate. The textural parameters were measured by a texture analyzer. Percentages of 7S, 11S, 11S+7S, A3 subunit and ratio of 11S/7S were calculated after Sodium Dodecyl Sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Phytic acid, Ca2+ and Mg2+ were determined. Pearson's correlation coefficients between seed components and tofu firmness were calculated. Results showed across locations over two years, A3 subunit content was strongly correlated with filled tofu firmness (r= 0.82, p<0.0001) and pressed tofu firmness (r= 0.83, p<0.0001). 7S content did not show any correlations with tofu firmness for both pressed tofu and filled tofu. 11S content correlated with filled tofu firmness (r=0.70, p<0.001) and pressed tofu firmness (r=0.53, p<0.01). No significant correlations were found between total (7S+11S) content and tofu firmness. 11S/7S ratio only exhibited moderate correlations with filled tofu firmness (r=0.47, p<0.05). For pressed tofu, firmness was found to negatively correlate with the tofu yield (r=-0.78, p<0.001). Locations and years showed no significant variations for tofu firmness. Phytic acid, Ca2+ and Mg2+ content did not significantly correlate with the firmness of tofu. The current study confirmed the validity of using A3 peptide as a biomarker in soybean breeding or as a criterion for estimating tofu firmness in both tofu manufacturing and food-grade soybean trade. The findings have been presented in American Chemical Society and in National Plant Breeder Association in 2019.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Chen, R., Chang, S. K. C., Zhang, Y., Gillen, A. M., Chen, P., Zhang, B. 2019. Relationships between protein composition and texture of tofu made from soybeans planted in three locations. American Chemical Society annual fall meetings. San Diego, CA. August 25-29, 2019
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Chen, R., Chang, S. K. C., Gillen, A. M., Chen, P., Zhang, B. 2019. Positive correlation between A3 subunit of glycinin and firmness of tofu made from soybeans grown in three locations over two years. National Association of Plant Breeders annual meeting. Pine Mountain, Georgia. August 25-29, 2019.
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Progress 03/15/18 to 03/14/19
Outputs
Target Audience:The audience includes soybean breeders, geneticists, soybean growers, food scientists, graduate students, and food and agricultural industries. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The project provided an opportunity for training a graduate student in food science at the Mississippi State University. The student has completed all course work successfully and learned all the laboratory skills and statistical analysis methods needed to conduct this research. How have the results been disseminated to communities of interest?The first year's results are expected to be disseminated to scientists in the agricultural and food chemistry field at the American Chemical Society meeting in the Fall of 2019. An abstract will be submitted in March of 2019 for presentation in the Fall. What do you plan to do during the next reporting period to accomplish the goals?We will analzye the 2018-planted soybeans for their compositions, protein profiles, and tofu quality, and carry out statistical analyses of the results of the combined data from both years. We will report the data in the next year's reporting period. The student is anticipated to write a master thesis on this subject. We will also submit our results for publication in a reputable agricultural or food chemistry journal. The results will be used to prepare for a full grant proposal to be submitted to the NIFA program in 2020.
Impacts
What was accomplished under these goals?
The impact of the results since the inception of the project in 2017 confirmed the hypothesized positive relationships existed between the targeted protein composition of the soybean and the tofu product texture properties. This confirmation was fostered by the fact that soybeans were grown in three different environments, including Mississippi, Missouri and Virginia. This is important as the positive relationship provides a solid foundation of genetics for the future development of improved food-grade soybeans for domestic industry and export to Asia for making tofu and possibly for other soybean protein products. The US-food grade soybean market in Asia for a long time has been eroded by the competition of soybeans from Canada. Improving the genetic traits to improve food quality of US soybean will increase the market size of US-grown soybean, and increase premiums for the growers. In addition, tofu with a better quality trait will be beneficial to the consumers. Twenty-two high-protein soybean cultivars and five check varieties were grown in three states. Their agronomic traits and yield were documented. After harvest, the soybeans were shipped to Mississippi State University for tofu quality testing and for protein profile composition analysis. Tofu was made from soybeans by two processing methods, namely the filled tofu and pressed tofu methods. The textural quality traits such as yield, firmness, cohesiveness, and brittleness were analyzed. The results from analyzing the profiles of the soy proteins using denaturing sodium-dodecyl-sulfate electrophoresis under reducing conditions showed that the ratios of glycinin to beta-conglycinin, and the specific A3 subunit peptides were significantly correlated with the tofu gel textural properties within each location and in the combined data of the three locations (probability level of less than 0.0001). Moreover, the relationships between A3 subunit and tofu firmness (r = 0.79 for both filled and pressed tofu) is much stronger than the ratios of glycinin to beta-conglycinins (r = 0.5 and 0.13 for filled and pressed tofu, respectively). The results validated our hypothesis that the expression of A3 protein subunit in high protein varieties is quantitatively related to the degree of firmness of the gel texture of tofu made by the two processing methods. The results implied that the A3 subunit is a critical component for the formation of the network structure of the tofu gels. There was also a significant correlation between the yield of pressed tofu and firmness of tofu. This result indicated the higher water-holding ability of the gels was derived from a higher firmness tofu soybean, which contained a higher A3 subunit. Other components such as the contents of proteins, calcium, magnesium have been determined for the 2017-harvested soybeans, and results are being analyzed for statistical correlations with the protein patterns and tofu quality. Calcium was found to have a significant correlation with the firmness of filled tofu. The harvest of the 2018-planted soybeans have been received from the three planting locations, and are undergoing testing for yearly (seasonal) effect. If the relationship between tofu quality traits and A3 subunit continues to hold from year to year, the results will further substantiate our research hypothesis. The project's time has been extended for one more year due to the need for more planting and analytical times. Our objectives for 2017-2018 were fully met. The overall project results will be reported in 2020.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2017
Citation:
Chang, S. K. C., Gillen, A., Chen, P. Y. and Zhang, B., Meng, S., and Tan, Y. Q. 2017. Enhancing protein composition in soybeans for improving tofu quality using A3 subunit as a marker. Annual Conference of the National Association of Plant Breeders. University of California-Davis. Aug 7-10, 2017.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Chang, S. K. C. 2018. The biochemistry of soybean for tofu making as affected by cultivars, processing and storage with an emphasis on protein chemistry. Innovations in Food Science and Nutrition. A keynote speech. Rome, Italy. September 13-15, 2018.
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Progress 03/15/17 to 03/14/18
Outputs
Target Audience:The project started in March of 2017. The audience includes food scientists, breeders, geneticists in university, USDA-ARS and the soy food industry. We have visited three soybean breeding and processing plants: the Cargill, Pioneer-hybride, and Bayers Soybean breeding programs in Missouri-Arkansas area. We also have presented a poster at the UC-Davis in the National Association of Plant Breeders, in that many scientists from academia and farming industries attended our posters. Changes/Problems:The soybean grown in Missouri was affected by Dicamba herbcide spray from the neighbor farms. The development of the plants were somewhat abnormal. However, the soybeans seemed to be able to be processed into tofu, and good correlations were obtained. Therefore, the problem is minor, but we need to be cautious during 2018 planting season. What opportunities for training and professional development has the project provided?This project provides an excellent opportunity for the collaboration between three universities and the USDA-ARS Crop Genetics unit in Stoneville to understand how environment affects the expression of the soybean proteins, which play an important role in determining the quality of tofu food. This is a two year project, and soybeans will be planted in the summer of 2018 to determine seasonal effect. Soybean researchers, graduate student, and technicians benefit from this project by learning and generating new knowledge. At the completion of the project, we anticipate that the results will generate data for the next phase of research to develop protein or gene marker for breeding to improve soybean and soy food quality. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?This is a two year project, and we plan to grow the same soybean varieties in the same three locations in the summer of 2018 for determining seasonal effect. After harvesting, soybeans will be cleaned up. The yield and traits of soybeans will be documented. We will then carry out chemical analyses, electrophoresis identification and quantification, and tofu making by the two methods which can be related to industrial processing of tofu. The same protocols as they are used in the first year will be used for determining the chemical and physical properties of soybeans and tofu.
Impacts
What was accomplished under these goals?
The results from this project is anticiapted to confirm the relationships betwee protein composition and tofu quality of soybeans grown in three locations. This will prove the environmental effect on gene expression, and ultimately the results will provide sound scientific foundation for the development of better quality cultivars for tofu making. Such high quality food-grade sobyeans will command a premium for soybean growers and compete favorably in the international market. Our specitif objective is to validate our hypothesis based on our preliminary findings that the expression of soybean A3 subunits in high protein soybean varieties is quantitatively related to the degree of firmness of tofu gel texture. In this first year, we have successfully hired a graduate student for this project. Twenty-two experimental soybean and 8 check cultivars were planted in May in duplicate plots in three different environments: states of Mississippi, Missouri, and Virginia. Soybeans were planted in May and harvested in the Fall of 2017. After cleaning, the yield and agronomic characteristics were documented. The yield from Stoneville, Mississippi was higher than that grown in Missouri and Virginia. After arrriving to our processing laboratory, we made filled tofu from the soybean samples, and conducted statistical analysis of the tofu firmness between growing locations and between the results from this year and the previous year (2014 grown in Stoneville, Mississippi). The firmness of filled tofu from the three locations ranged from 80 to 157 g per cm square of the tofu gel. We found that the tofu firmness of the cultivar MS-1 consistently showed the highest tofu firmness in all three locations among all varieties. This was also consistent with our preliminary study. Pearson correlation analysis among all locations and with previous 2014 study indicated that the firmness of filled tofu was strongly correlated among locations with correlation coefficients ranging from 0.61 to 0.82, highly significant at the probability level of 0.0001. The correlation coefficient between the average firmness values of the three locations and the 2014 preliminary study was 0.78 with probability level of 0.0001. Thus far, the results showed consistent gene expression of the soybean proteins in 3 growing locations even though there were differences among locations. This was expected. Currently, we are analyzing the electrophoresis patterns of the soybeans, and will report the results in this year's Project Director's meeting and next year's CRIS report.
Publications
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