WAXY BARLEY STARCH AS RAW MATERIAL FOR HEALTHY FOOD PRODUCTS

ductive one, the number of productive nodes and pods per node, paying attention not only to their total number and pairing, but also to the presence and plumpness of beans on the upper productive nodes. We also measure the seed number per pod. In F 3 , offspring is assessed for evenness and the vegetation phase lengths. In F 4 - 5 , only even high-yielding families are selected; they are threshed; and the total number of seeds is estimated. General requirements for the selection of hybrid and breeding material depend only on the direction of breeding. Conclusions. The application of the pedigree method in pea breeding is time-consuming for breeders and demands thoroughness at all stages. However, this approach allows detailed in-vestigating hybrid material at the beginning of the breeding process, which increases the breeding effectiveness. The effectiveness of the applied methods and selection ways is con-firmed by demand for pea varieties bred at the Plant Production Institute named after VYa Yuriev and positive feedback from manufacturers. The article justifies the expediency of the breeding of barley cultivars with waxy starch, grain of which can be raw material for functional food production. The study purpose was to create food barley cultivars with waxy starch. To accomplish this, qualitative parameters of barley accessions were determined, and starting material was selected for breeding. The study was conducted at the Plant Production Institute named after V.Ya. Yuriev of NAAS in 2015–2017. Twenty four chaffy and naked barley accessions of different origin were taken as the test material. Grain quality parameters of wild type and waxy accessions were compared. The yield, protein and starch contents, endosperm vitreousness, and antioxidant activity were shown to depend both on growing conditions and on genotypes. High nutritional quality of waxy barley cultivars was proved, videlicet, significantly higher content of oil compared with accessions with wild type starch and lower vitreousness of endosperm in chaffy accessions. Accessions with very valuable characteristics were selected: line 12-1014 with high digestibility of protein (61.75 mg tyrosine/g protein for pepsinolysis + trypsinolysis), line 12-945 with high antioxidant activity (2.02 mg chlorogenic acid equivalent/g), line 12-954 with a high content of oil (3.75%) and awnless line 14-1183 with a high content of polyunsaturated ω-3 linolenic acid (6.09%). These lines are valuable both as starting material for the breeding of food cultivars, and directly for functional food production. Waxy cultivars Shedevr and Amil were submitted to the state variety trials. Grain of these cultivars is noticeable for high contents of starch ( 60%), oil (3.45–3.47%) and fiber (3.65– 3.70%). Antioxidant activity is also sufficiently high (1.94–2.07 mg chlorogenic acid equiva-lent/g). Being low vitreous (40–43%), cultivars Shedevr and Amil can be used for production of flakes and flour, including extruded flour. Purpose and objective . The study purpose was to create waxy barley cultivars suitable for functional food production. For this purpose, quality parameters of barley accessions were determined and starting material was selected for breeding. Materials and methods . The study was conducted at the Plant Production Institute named after V.Ya. Yuriev of NAAS in 2015–2017. Twenty four chaffy and naked barley accessions of different origin with waxy and wild type starch were taken as the test material. The data were statistically processed by analysis of variance; a posteriori comparison was performed using homogeneous groups (Fisher LSD test) in software STATISTICA 10. Relationships between traits were evaluated using correlation and regression coefficients. The protein and starch contents were measured on an InfraLUM FT-10M 09495, vitreousness was determined on a diaphanoscope. Waxy genotypes were identified by dyeing with Lugol‟s iodine solution (modified Juliano's method). The antioxidant activity (AOA) of barley accessions was evaluated by capacity of ethanol extracts to neutralize the DPPH (2,2-diphenyl-1-picyrylhydrazyl) radical. The antiradical activity was determined as described by S. Arabshahi and A. Urooj. The in vitro ability of protein to be digested by proteolytic enzymes was determined by O. Pokrovsky and I. Ertanov‟s method, which consists in gradual action of proteinases (crystalline pepsin and trypsin) on proteins. Results and discussion . Grain quality parameters of wild type and waxy accessions were compared. The yield, protein and starch contents, endosperm vitreousness, and antioxidant activity were shown to depend both on growing conditions and on genotypes. High nutritional quality of waxy barley cultivars was proved, videlicet, significantly higher content of oil compared with accessions with wild type starch and lower vitreousness of endosperm in chaffy accessions. Accessions with very valuable characteristics were selected: line 12-1014 with high digestibility of protein (61.75 mg tyrosine/g protein for pepsinolysis + trypsinolysis), line 12-945 with high antioxidant activity (2.02 mg chlorogenic acid equivalent/g), line 12-954 with a high content of oil (3.75%) and awnless line 14-1183 with a high content of polyunsaturated ω-3 linolenic acid (6.09%). These lines are valuable both as starting material for the breeding of food cultivars, and directly for functional food production. Waxy cultivars Shedevr and Amil were submitted to the state variety trials. Grain of these cultivars is noticeable for high contents of starch ( 60%), oil (3.45–3.47%) and fiber (3.65–3.70%). Antioxidant activity is also sufficiently high (1.94–2.07 mg chlorogenic acid equivalent/g). Being low vitreous (40– 43%), cultivars Shedevr and Amil can be used for production of flakes and flour, including extruded flour. Conclusions . Thus, the high nutritional value of barley varieties with modified starch composition was proven. High-nutrient lines, which are valuable both as starting material for the breeding of food varieties and directly for the production of functional foods, were distin-guished. Varieties Shedevr and Amil were submitted to the State variety trials, and Shedevr been entered in the State Register of Plant Varieties Suitable for Dissemination in Ukraine

amylopectin, while the correlation between the protein content and the percentage of long chains of amylose was negative [23].
The Department of Genetic Foundations of Breeding of the Plant Breeding and Genetics Institute of NAAS created breeding lines with high performance, including waxy ones, with technological parameters of grain suitable for production of a wide range of products: with hard grain for production of flour, groats, noodles, pasta and with soft grain for production of flakes and drinks. Commercial Canadian naked cultivars Alamo and Candle served as sources of the wax gene [2,24].
The Plant Production Institute named after V.Ya. Yuriev of NAAS breeds waxy barley with lines UA 0804955 (Belarus), UA 039699, UA 039701, U.S. 039748 (USA), and IR 6912 (Mexico) as the sources of the wax gene. Other breeding components were domestic cultivars. As a result, high-performance and adapted to local growing conditions lines with waxy starch were created [25,26].
Barley is used in healthy food production due to its positive effects on human health [27]. Oxidative stress and inflammation are two key factors that contribute to atherosclerosis development. Studies of barley, in particular, of its protective function in regulation of antioxidant protection, revealed powerful protective features of barley in oxidative processes and the potential role of barley in preventing chronic inflammation in cardiovascular diseases [28]. Japanese researchers reported that consumption of waxy barley groats improved the intestine functioning, especially in the elderly [29].
Food products made from barley grain or flour with various amylopectin / amylose ratios considerably differ in technological properties and quality. Products made from waxy barley are characterized by reduced hardness compared to products made from wild type barley. Tortillas from waxy barley are better rolled and less fragile [17].
Thus, the starch properties have a significant impact on the food quality, and creation of barley cultivars with modified amilopectin / amylose ratios is essential for healthy food production.
Purpose and objective. The study purpose was to create waxy barley cultivars suitable for functional food production. For this purpose, quality parameters of barley accessions were determined and starting material was selected for breeding.
Materials and methods. The study was conducted at the Plant Production Institute named after V.Ya. Yuriev of NAAS. Twenty four chaffy and naked barley accessions of different origin with waxy and wild type starch were taken as the test material. Cultivar Vzirets, which is the best in the zone of our invеstigations, was taken as the standard.
The data were statistically processed by analysis of variance; a posteriori comparison was performed using homogeneous groups (Fisher LSD test) in software STATISTICA 10. Relationships between traits were evaluated using correlation and regression coefficients.
The protein and starch contents were measured on an InfraLUM FT-10M 09495. vitreousness was determined on a diaphanoscope. Waxy genotypes were identified by dyeing with Lugol"s iodine solution (modified Juliano's method). Starch granules of waxy barley are discolored to dark red or brown, and wild type starch granules -to dark-blue [30].
The antioxidant activity (AOA) of barley accessions was evaluated by capacity of ethanol extracts to neutralize the DPPH (2,2-diphenyl-1-picyrylhydrazyl) radical. The method is based on discoloration of ethanol solution of the stable DPPH• in the presence of ethanol extracts of grain products containing antioxidants that are proton donors capable of neutralizing the DPPH radical and translate it into a decolorized reduced form. The antiradical activity was determined as described by S. Arabshahi and A. Urooj [31].
The in vitro ability of protein to be digested by proteolytic enzymes was determined by O. Pokrovsky and I. Ertanov"s method, which consists in gradual action of proteinases (crystalline pepsin and trypsin) on proteins [32].
Results and discussion. Weather conditions are important to obtain top-quality raw materials. For example, there is evidence that high temperature during ear formation increases the protein content and decreases the starch content in grain, depending on the genotype. There is also evidence that waxy genotypes are more responsive to high temperatures than wild type ones [33].
The weather conditions during the study period were various; however, they were favorable for the growth and development of barley plants in the "emergence-tillering" phase. In 2015, there was a drought in the crucial phases of earing and grain filling, and in 2016 an excessive amount of precipitation caused plant lodging during this period (Table 1). The yields of barley accessions were the highest in 2017 and the lowest in 2015, which is accounted for the weather conditions during the "ear formation" and "grain flilling" phases, however, no significant difference in the yields was noticed over the years. The performance of the naked cultivars significantly differed from that of chaffy ones, both in the group with wild type starch and in the waxy group ( Table 2). The lower yield of naked cultivars is due to the absence of glume, which makes up 8-10% of the grain weight. The yields of waxy accessions were lower than those in the group with wild type starch, but this difference was not significant. Thus, in our study, the yields of waxy accessions are similar to those of accessions with wild type starch, which is at odds with other researchers" results [22]. Grain quality for food production is primarily determined by protein and starch contents. Proteins are extremely important for the human body, since they belong to essential irreplaceable substances. In our study, the protein content was significantly higher in naked cultivars both with waxy (14.30%) and wild type (14.03%) starch (Table 3). There was no significant difference in the protein content between chaffy cultivars with wild type and waxy starch (12.41% and 12.34%. respectively). In other studies [22,23], the protein content was significantly higher in all waxy accessions. i.e. our data are consistent with these results.
There was no significant difference in the starch content between the groups of accessions (59.52%, 60.32%, 60.80%, 59.53%) (see Table 3). Depending on the year conditions. the starch content was significantly higher in 2016 compared with the other years. This is in line with the literature data indicating that low temperature during "ear formation-grain filling" period increases the starch content [33]. In addition to protein content in foods, one should know the level of protein absorption by the human body. This parameter is determined by the biological value of protein, which depends on the presence of essential amino acids, their ratio to nonessential ones and digestibility in the gastrointestinal tract. Enzymatic hydrolysis can be carried out in conditions close to the conditions of digestion in a living organism, which is a biochemical method for assessing protein digestibility. In this case, in vitro attackability of proteins is determined, which gives the complex characterization of the protein under investigation compared to the reference (casein). We distinguished waxy line 12-1014 by the amount of soluble products of pepsinolysis and trypsinolysis (61.75 mg tyrosine/g protein). Other lines and cultivars had significantly lower digestibility than that of line 12-1014 (Table 4).
According to the current requirements to food barley cultivars, they are expected to have vitreous endosperm. Vitreousness is an external trait of the grain quality; it reflects the structure of internal tissues of grain. In ripe barley grain, starchy endosperm cells are filled with starch granules immersed in the protein matrix. For floury endosperm, weak bonds between starch grainules and proteins are intrinsic. In vitreous endosperm, such bonds are very strong. Vitreousness is influenced by growing conditions during the "grain filling" and "ripening" phases: the more frequent dry days are, the higher this index is. Vitreous grain usually contains more protein than floury one. In our study, the vitreousness of chaffy cultivars was significantly lower (49%, 51%) than that of naked ones (94%, 88%). At the same time, waxy cultivars had a lower vitreousness than wild type cultivars, but this difference was insignificant (Table 5). Thus, having soft grain, waxy accessions are suitable for production of flour, including extruded flour, and flakes. The effects of the growing conditions during the crucial phases of vegetation on endosperm vitreousness in spring barley cultivars were investigated. The precipitation during the "ear formation -grain filling" period reduced vitreousness, especially in naked cultivars (r = -0.846). There was only a positive tendency between the sum of effective temperatures in the "ear formation -grain filling" phase and vitreousness in chaffy cultivars, while in naked ones there was a significant positive correlation ( r = 0.803).
Rise in temperature in the "grain filling -ripening" phase significantly increases endosperm vitreousness, with the correlation coefficient ranging within 0.526-0.875. The dependence of vitreousness on the precipitation amount in the "grain filling -ripening" phase is insignificant, since a negative tendency was only observed. This is accounted for the fact that in the research area precipitation is usually very rare and showery during the "grain filling -ripening" period in barley, which can not significantly affect the parameters under investigation.
The antioxidant activity is a major factor determining the value of food for human health. The level of this parameter in barley is one of the highest among cereals. We found that the antioxidant activity depended both on the genotype and on the conditions of cultivation and analysis type. Thus, the highest AOA was recorded in 2016, the lowest -in 2015 (Table 6). At the same time, there was a steady dependence of the AOA on the genotype, which is consistent with numerous data of other researchers [13,14,15]. Under any conditions, the AOA was the highest in naked genotypes with waxy starch. This parameter had no significant differences within the other groups of accessions, with the lowest values in chaffy accessions with wild type starch (see Table 6). It should be noted that among all the accessions the highest AOA was in naked cultivar Alamo (up to 3.46 mg chlorogenic acid equivalent/g), which is considered the reference for this parameter in the world.
According to the literature data, genetic factors that control the waxy trait can be associated with genetic factors that increase the lipid content in barley [12,34]. Our results are consistent with these data. The oil content in grain of waxy accessions is significantly higher (2.92-3.52%) than that both in chaffy (2.55%) and in naked (2.53%) accessions with wild type starch (Table 7). Among the waxy accessions, the oil content is significantly higher in chaffy accessions (3.52%) compared to naked ones (2.92%).
After burning barley grain, organic carbon-free ash, which is a concentrate of grain minerals, remains. We found that the ash content was, expectedly, significantly higher in chaffy accessions. This parameter does not depend on starch composition. The fiber content is of great importance for the digestive processes. Total fiber, as a complex of nutritional value, consists of insoluble and soluble components. We revealed that the average content of fiber in waxy accessions was higher than that in wild type ones, but the difference was insignificant (Table 7). This is only partially consistent with other researchers" results [12,35,36].
As a part of vegetable oils, including barley oil, unsaturated fatty acids are very important for healthy human nutrition. Barley oil is different from others by high levels of such acids, in particular, of polyunsaturated ω-3 linolenic acid. In our study, the linolenic acid content was the highest in chaffy cultivars with wild type starch bred at the Plant Production Institute nd. a V.Ya. Yuriev of NAAS: in Lider (6.64%), Krasen (6.52%), Vzirets (6.50%), and Ahrarii (6.30%). Of the waxy barley lines and cultivars, the highest contents of polyunsaturated fatty acids were found in cultivar Shedevr (linolenic acid 5.85%, linoleic acid 54.68%), awnless line 14-1183 (linolenic acid 6.09%), and naked cultivar Candle (linoleic acid 54.80%) ( Table 8). The waxy lines with the best quality and economic parameters were submitted to the state variety trials: lines 12-473 and 12-476 called Shedevr and Amil, respectively. The yields of these lines amount to 5.38-5.43 t/ha, which is higher by 8-12% than that of standard Vzirets. The protein content is medium; the starch content is high ( 60%) ( Table 9). The oil content (3.45-3.47%) and fiber content (3.65-3.70%) are high. The antioxidant activity is also sufficiently high (1.94-2.07 mg chlorogenic acid equivalent/g). Grain of cultivars Shedevr and Amil is characterized by low vitreousness (40-43%), therefore, it is suitable for production of flakes and flour, including extruded flour.
The lines of high nutritional quality of grain are used in breeding as starting material for hybridization. Cultivars Shedevr and Amil were submitted to the state variety trials. Conclusions. Thus, as a result of the study, high nutritional qualities of grain of barley waxy cultivars were demonstrated. In particular, such barley genotypes have significantly higher oil contents than wild type accessions, and their endosperm is less vitreous in chaffy accessions. Several accessions with very valuable characteristics were selected: line 12-1014 with a high protein digestibility (61.75 mg tyrosine/g protein for pepsinolysis + trypsinolysis), line 12-945 with a high antioxidant activity (2.02 mg chlorogenic acid equivalent/g). Line 12-954 was singled out due to the oil content (3.75%); awnless line 14-1183 -due to a high content of polyunsaturated ω-3 linolenic acid in oil (6.09%). Such lines are valuable both as starting material for the breeding of food cultivars and directly for functional food production. Having appropriate economic and qualitative parameters, cultivars Shedevr and Amil can be used for production of flakes and flour. В статье обоснована целесообразность селекции сортов ячменя с waxу-крахмалом, зерно которых может быть сырьем для производства продуктов функционального питания. Цель исследования. Целью исследования было создание пищевых сортов ячменя с waxукрахмалом. Для этого были определены показатели качества образцов ячменя и выделен исходный материал для селекции. Материалы и методы. Исследования проведены в Институте растениеводства им. В.Я.