Dr. Robert Yaklich
Plant Physiologist,
USDA-ARS Soybean and Alfalfa Research Laboratory,
Beltsville, MD 20705
Tel. 301-504-5723
FAX 301-504-5728
Introduction:
Soybeans are an important agricultural crop because of the oil and protein contained in the seed. The seed contains about 20% oil and 40% protein. The oil is used mainly for human consumption in products as cooking oil, margarine, and salad dressings. After the oil is extracted from the seed the material that is left is the soybean meal. The soybean meal is an excellent protein source for animal husbandry and is an important component in the feeding ration in the poultry and hog industries. In many countries, the soybean is an important component of the human diet. Soybeans are a leguminous plant. What this means is that through a companionship of the plant roots and beneficial bacteria they fix atmospheric nitrogen to supply some of the plants nitrogen needs. Other common plants that are also legumes are the bean, pea, and peanut. All of these plants can fix atmospheric nitrogen to supply some of their nitrogen needs. Nitrogen is used by plants and animals to form different nitrogen containing compounds found in all living organisms. One group of these compounds are called amino acids that are used by biological organisms to make proteins. There are different types of proteins in living organisms that have different functions. My research deals with proteins that are found in seeds and specifically seed storage proteins.
Soybean seed proteins:
The soybean seed contains special cells that make and store the seed storage proteins. Once the proteins are formed they are stored in special structures within the cells that are called protein storage vacuoles (Figure 1). The pictures in Figure 1 are called micrographs and were obtained by viewing very thin sections of soybean seed in a transmission electron micrograph. The protein storage vacuoles differ in size and the protein is made and stored during the time the seed is growing on the plant. The top picture is from a young seed and the dark portion represents protein that is being stored in the protein storage vacuole. As the seed grows the vacuoles fill with more storage protein and become full as shown in the bottom picture.
Figure 1. Transmission electron micrographs of protein storage vacuoles in developing soybean cotyledon parenchyma cells. (Top) Parenchyma cells observed in a young developing seed. (Bottom) Parenchyma cells observed in a mature developing seed. Figure 2. Low temperature scanning electron micrographs of protein storage vacuoles in developing soybean cotyledon parenchyma cells. (Top) Parenchyma cells observed in a young developing seed. (Bottom) Parenchyma cells observed in a mature developing seed.
An example of how the cells and protein storage vacuoles are arranged is shown in Figure 2. These are pictures of developing soybean seed that were observed with another type of electron microscopy. This electron microscope is a scanning electron microscope and is used to look at the surface structure. These pictures were taken with a special instrument because it looks at frozen tissue and is called low temperature scanning electron microscopy. There are many cells in the soybean seed and the round structures are protein storage vacuoles. The picture shows the protein storage vacuoles in a young developing seed (top) and another (bottom) that is close to maturity. The seed makes thousands of these protein storage vacuoles to store the seed storage protein. Luckily, there are only a few major seed storage proteins in soybean. Two storage proteins, B-conglycinin and glycinin, account for approximately 70% of the protein stored in the protein storage vacuoles. Generally, glycinin is the more abundant of the two storage proteins.The amount of these two storage proteins will differ in soybean seeds depending on their genetics and where they are grown. Where the application of a protein product calls for more sulfur amino acids then a higher glycinin content is preferred. The ratio of these two storage proteins is also important in the manufacture of food products. Changes in the B-conglycinin:glycinin ratio result in differences in the physical properties of tofu. Tofu containing more B-conglycinin is softer than tofu from soybean containing more glycinin. Recently, the protein fraction of soybean seeds has become more important than that of the oil. This is mainly due to an increase in the standard of living throughout the world and the need for more protein for animal rations. Soybean breeders have made new protein lines that exceed 50% protein. This has been relatively easy to do and also means that there may be some simple factor that allows the soybean seed to make and store more protein in the protein storage vacuoles. The kind and amount of storage protein increased in these new soybean lines is not known. The previous two examples demonstrate that knowing the storage protein content is important. We are attempting to determine which storage proteins are increased by looking at the quantity of B- conglycinin and glycinin in these lines. This is being done by analyzing their subunit structure by gel electrophoresis (Figure 3).
Figure 3. Polyacrylamide gel of some high-protein soybean lines. The principal protein subunits of B-conglycinin are the a', a, and B subunits and the principal polypeptides of glycinin are the acidic A3, A1,2,4 and basic polypeptides.
In gel electrophoresis, proteins are separated by their size and charge. The protein bands are stained to make them visible. The proteins at the top of the gel are larger than those at the bottom. The gel shows the a', a, and b subunit protein bands that make up the B-conglycinin storage protein. The gel also shows the acidic and basic polypeptides that make up the glycinin storage protein. We have found differences in the content of the components of B- conglycinin and glycinin in the high protein lines. This means there may be several mechanisms regulating the amount of these two storage proteins that are synthesized and stored in the high protein lines. This information may be used to selectively increase the type of storage protein for specific uses. Soybeans used in feeding rations for the hog industry would be better served with soybeans containing more B-conglycinin and the soybeans used in the poultry industry would be better served with soybeans containing more glycinin. Future research will be directed towards regulation of the quantity of B-conglycinin and glycinin in the soybean seed.
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