Rice varieties with similar amylose content differ in starch digestibility and glycemic response in humans.

Three high-amylose rice varieties, IR42, IR36, and IR62, with similar chemical composition including amylose content (26.7-27.0%), were cooked under the same conditions and tested for in vitro digestibility as well as blood glucose and insulin responses in healthy human volunteers. The starch-digestion rate and the glycemic and insulin responses were the highest in IR42, followed by IR36 and, then IR62. The differences were not due to unabsorbed carbohydrate but were related to their physicochemical properties, such as gelatinization temperature, minimum cooking time, amylograph consistency, and volume expansion upon cooking. When the three varieties were cooked for their minimum cooking time, they had the same degree of gelatinization and their starch-digestion rates and glycemic responses were similar. We conclude that amylose content alone is not a good predictor of starch-digestion rate or glycemic response. Rice varieties with similar high-amylose contents can differ in physicochemical (gelatinization) properties and this, in turn, can influence starch digestibility and blood glucose response.

Aspects of nitrogen metabolism in the rice seedling.

The effects of nitrogen source NO(3) (-) or NH(4) (+) on nitrogen metabolism during the first 2 weeks of germination of the rice seedling (Oryza sativa L., var. IR22) grown in nutrient solution containing 40 mug/ml N were studied. Total, soluble protein, and free amino N levels were higher in the NH(4) (+)-grown seedling, particularly during the 1st week of germination. Asparagine accounted for most of the difference in free amino acid level, in both the root and the shoot. Nitrate and nitrite reductase activities were present mainly in the shoot and were higher in the NO(3) (-)-grown seedling, whereas the activity of glutamate dehydrogenase and glutamine synthetase in the root tended to be lower than that of the NH(4) (+)-grown seedling during the 1st week of germination. Glycolate oxidase and catalase activities were present mainly in the shoot. Maximum activity of the above five enzymes occurred 7 to 10 days after germination. Differences in the zymograms of nitrate reductase, glutamate dehydrogenase, and catalase were mainly between shoot and root and not from N source. Nitrite reductase bands were observed only in plants grown in plants grown in NO(3) (-).Ten-day-old seedlings of three rices differing in level of grain protein did not differ in the level of N fractions and of enzyme activities, which were consistent with their differences in grain protein content.

Enzymic degradiation of starch granules in the cotyledons of germinating peas.

Starch, total alpha- and beta-amylase, and phosphorylase levels and the zymogram patterns of these 3 starch-degrading enzymes were determined in the cotyledons of smooth pea (Pisum sativum L.) during the first 15 days of germination. Starch is degraded slowly in the first 6 days; during this time, alpha-amylase is very low, beta-amylase is present at a constant level while phosphorylase gradually increases and reaches a peak on the fifth day. Beginning on the sixth day there is a more rapid degradation of starch which coincides with alpha-amylase production. One phosphorylase band and 2 beta-amylase bands are present in the zymogram of the imbibed cotyledon. An additional phosphorylase band and 1 alpha-amylase band appear during germination. Seeds imbibed in benzyladenine, chloramphenicol, and in cycloheximide show retarded growth and slower starch degradation and enzyme production than the controls. We conclude that alpha-amylase is the major enzyme involved in the initial degradation of starch into more soluble forms while phosphorylase and beta-amylase assist in the further conversion to free sugars.

Biochemical Changes in the Rice Grain during Germination.

Changes in the content of starch, protein, and RNA and in the activity of their hydrolases in the rice endosperm (Oryza sativa L., variety IR8) were determined during the first week of germination without added nutrient both in the dark and in the light. Changes were generally more rapid in the dark than in the light. Oxygen uptake and RNase activity started to increase and the root protruded on the second day, followed by the coleoptile on the third day, and the primary leaf on the fourth day. ATP level was at a maximum on the fourth day. The activity of amylases and R enzyme increased progressively, but that of phosphorylase tended to decrease during starch degradation. A new alpha amylase isozyme band appeared during germination. Glucose was the major product of starch degradation. Sucrose, maltose, maltotriose, raffinose, and fructose were also detected. Protease activity reached a maximum on the fifth or sixth day and closely paralleled the increase in soluble amino N and soluble protein.In embryoless seed halves with 0.12 muM gibberellin As, peak protease activity occurred in 2.5 days and peak alpha amylase activity on the fifth day of incubation. The production of alpha amylase, protease, and R enzyme was inhibited by 40 muM cycloheximide, but only alpha amylase and R enzyme were inhibited by 20 mug/ml actinomycin D.

Changes in the Activity of Some Hydrolases, Peroxidase, and Catalase in the Rice Seed during Germination.

A study was made of the changes in activity of enzymes involved in the breakdown of stored phytin, lipid, and hemicellulose in the aleurone layer of rice seed (Oryza sativa L., variety IR8) during the 1st week of germination in the light. Enzyme assays were made on crude extracts from degermed seed, and activities were expressed on a per seed basis. Phytase activity increased within the 1st day of germination. The increase in activity of most other enzymes-phosphomonoesterase, phosphodiesterase, esterase, lipase, peroxidase, catalase, beta-glucosidase, and alpha- and beta-galactosidase-closely followed the increase in protein content. Their peak activities occurred by the 5th to the 7th day. Some enzymes, such as beta-1, 3-glucanase and alpha-amylase, continued to increase in activity after the 7th day. Phytase, beta-1, 3-glucanase, and alpha-amylase followed a similar sequence of production in embryoless seed halves incubated in 0.12 muM gibberellin A(3), but the production of lipase was delayed.

Some properties of 3-phosphoglycerate phosphatase from developing rice grain.

Some properties of 3-P-glycerate phosphatase from developing caryopsis of rice (Oryza sativa L., variety IR26) were studied. The enzyme was found to be soluble and not bound to starch, and concentrated mainly in the pericarp-aleurone layer; its maximum activity was at 12 to 14 days after flowering. Contents of 3-P-glycerate and chlorophyll were highest in the grain at 7 to 8 days after flowering when starch synthesis was at a maximum. The enzyme was purified about 100-fold by precipitation with 50 to 80% ammonium sulfate, followed by chromatography through Sephadex G-200 and CM-Sephadex C-50. The pH optimum was from 5.7 to 6 and no cation was required for activity. The purified preparation had an apparent Km of 2.85 mm and was inhibited by Cu(2+), Hg(2+), Zn(2+), Fe(3+), molybdate, and F(-). The enzyme also exhibited high activity toward UTP, ATP, and p-nitrophenyl phosphate; moderate activity toward other phosphates; but no activity toward phytate. A molecular weight of about 23,000 was obtained for the 3-P-glycerate peak during gel filtration on Sephadex G-200, which corresponded to a value of 26,000 for the major protein fraction by thin layer gel filtration on Sephadex G-150. Zymograms of the whole extract and semipurified preparations showed two phosphatase bands with 3-P-glycerate as substrate.

Free sugars in relation to starch accumulation in developing rice grain.

The changes in sugars (water-soluble carbohydrates) were studied in the developing grain of rice (Oryza sativa L., variety IR28 and IR29) in relation to the role of these sugars as precursors of ADP glucose in starch accumulation. The levels of total sugars, total reducing sugars and free glucose, sucrose and other nonreducing sugars, maltooligosaccharides, and total and nonsucrosyl fructose followed closely the changes in the rate of starch accumulation, in both IR28 and 29; the peak value occurred 9 days after flowering. The level of soluble carbohydrates remained high in the caryopsis and also in milled rice after starch accumulation, suggesting that the supply of sugar precursors does not limit starch accumulation in the rice grain. Because of a higher level of reducing sugars, the level of free sugars in the grain of waxy rice IR29 was higher than that of nonwaxy IR28.

Variability in contents of thiamine and riboflavin in brown rice, crude oil in brown rice and bran-polish, and silicon in hull of IR rices.

Among 30 IR varieties, variety had greater effect on thiamine content of brown rice than season. Both season and variety affected riboflavin content. Purple- and red-pericarped Philippines rices had higher riboflavin and crude protein contents than IR rices. Crude oil contents of brown rice and bran-polish of 19 IR rices were significantly affected by variety, but only brown rice oil content was significantly affected by season. Bran-polish of waxy rice IR29 had the highest oil content. Silicon content of hull of 16 IR rices showed significant variation due to variety and season and tended to be higher in the dry season crop. Variety rankings in nutrient composition were not consistent for the two crops. The levels of nutrients of IR rices were within the range of values reported for other rices.

Protein and lysine digestibility and protein quality of cooked Filipino rice diets and milled rice in growing rats.

The protein quality of typical rice-based menu of Filipino preschool child and adult and cooked milled rice was assessed for true digestibility (TD), biological value (BV) and net protein utilization (NPU) in growing rats. Lysine and energy digestibilities were also determined. For the preschool child diet, TD was 88.8%, BV 90.0%, and NPU 79.9%. For the adult diet, TD was 87.3%, BV 86.6%, and NPU 75.5%, whereas cooked rice had 90.0% TD, 82.5% BV and 74.3% NPU. Lysine digestibility was 95.4% for preschool child diet, 95.7% for adult diet, and 100.0% for rice. Digestible energy was 91.3% for preschool child diet, 93.0% for adult diet, and 95.3% in rice. Amino acid scores were 100.0% for preschooler diet, 92.1% for adult diet, and 62.2% for rice. Protein quality based on amino acid score corrected for TD was 88.8% for preschool child diet. 80.4% for adult diet, and 56.0% for rice. If based on lysine digestibility instead of TD, protein quality would be 7.1% higher.

Biochemical factors affecting protein accumulation in the rice grain.

Rice grains (Oryza sativa L.) from three varieties and three pairs of lines with different protein content were collected at 4-day intervals from 4 to 32 days after flowering. The samples were analyzed for protein, free amino nitrogen, ribonucleic acid, protease activity, and ribonuclease activity. In addition, the capacity of the intact grain to incorporate amino acids was determined for the three pairs of lines. The maximal level of free amino nitrogen and the capacity of the developing grain to incorporate amino acids were consistently found to be higher in the samples with the high protein content in the mature grain. The ribonucleic acid content of the grain tended to be higher in the high protein samples.

Protein quality of high-protein and low-protein milled rices in preschool children.

High-protein (11.0%, IR480-5-9) and low-protein (7.1%, IR32) milled rices were compared with casein as a control when fed as the main source of dietary N to eight apparently normal Filipino children, 1.5 to 2.0 years of age. Daily diets were formulated to contain 250 mg N and 100 kcal/kg body weight. Apparent digestibility of 60.0% for high-protein rice was lower than for low-protein rice (66.2%), and higher values of 76.8 and 80.8% were obtained for the two casein control diets. Corresponding apparent retentions were similar for the two rices, at 23.4 and 26.9%, respectively, as compared to 25.0 and 38.6% for the two casein control diets. These results support previous data on children fed non-isonitrogenous rice-based diets indicating that an increase in protein content of milled rice results in only a slight decrease in its protein quality. The results were similar to those obtained with Peruvian children on a similar protocol.

Grain quality of rices grown in irrigated and upland cultures.

Upland culture of nine varieties at Ivory Coast resulted in decreases in grain weight, brown and milled rice translucency, Chroma meter L* of rough rice, Chroma meter L* and b* of milled rice, milled rice protein, and Amylograph peak viscosity, and in increases in total phenolics of hull and brown rice, and Chroma meter a* of rough, brown, and milled rice of 5 to 8 of the varieties as compared with irrigated culture. Upland culture had no consistent effect on grain resistance to Angoumois grain moth (Sitotraga cerealella).

Genetic analysis of waxy locus in rice (Oryza sativa L.).

Inheritance of waxy locus was studied in crosses of a waxy variety with four non-waxy parents having high-, intermediate-, low- or very low-amylose content. The analysis for amylose content was done on a single grain basis in parents, F1, F2, B1F1, and B2F1 seeds. The waxy parent lacking synthesis of amylose content was found to differ from the ones having high-, intermediate-, low- or very low-amylose content by one gene with major effect. Dosage effects for amylose content were observed to have great influence on segregation pattern and efficiency of selection. Selection efficiency for amylose content can be enhanced by selecting for endosperm appearance in early segregating generations.

Classification of cultivated rices into indica and japonica types by the isozyme, RFLP and two milled-rice methods.

Four methods for classifying cultivated rices (Oryza sativa L.) (including IR varieties) into indica and japonica types - waxy gene product in endosperm starch, glutelin α3 molecular weight in milled rice, RFLP polymorphism at the Wx locus and Glaszmann's isozyme method - were compared. On the basis of the two endosperm traits and the RFLP method Glaszmann's group 1 (indica) was classified as mainly indica and intermediate groups 2, 3 and 4 as exclusively indica. However, the endosperm traits classified Glaszmann's group 5 as mainly indica, while the RFLP method classified it as japonica. The RFLP waxy gene probe was closest to the isozyme method in classifying group 6 as japonicas; the waxy gene product gave mainly indica reaction even in group 6, and the glutelin α3 method was intermediate. All IR rices were classified as being indica on the basis of Wx gene product and by Glaszmann's method, but a few were classified as japonica by the glutelin α3 method and by the RFLP waxy gene probe.

Enzymes of starch metabolism in the developing rice grain.

The levels of starch, soluble sugars, protein, and enzymes involved in starch metabolism-alpha-amylase, beta-amylase, phosphorylase, Q-enzyme, R-enzyme, and starch synthetase -were assayed in dehulled developing rice grains (Oryzasativa L., variety IR8). Phosphorylase, Q-enzyme, and R-enzyme had peak activities 10 days after flowering, whereas alpha- and beta-amylases had maximal activities 14 days after flowering. Starch synthetase bound to the starch granule increased in activity up to 21 days after flowering. These enzymes (except the starch synthetases) were also detected by polyacrylamide gel electrophoresis. Their activity in grains at the midmilky stage (8-10 days after flowering) was determined in five pairs of lines with low and high amylose content from different crosses. The samples had similar levels of amylases, phosphorylase, R-enzyme, and Q-enzyme. The samples consistently differed in their levels of starch synthetase bound to the starch granule, which was proportional to amylose content. Granule-bound starch synthetase may be responsible for the integrity of amylose in the developing starch granule.

Starch metabolism in the leaf sheaths and culm of rice.

The levels of starch and dextrin, free sugars, soluble protein, and enzymes involved in starch metabolism-alpha-amylase, beta-amylase, phosphorylase, Q-enzyme, R-enzyme, and ADP-glucose starch synthetases-were assayed in the leaf sheaths and culm of the rice plant (Oryza sativa L., variety IR8) during growth.Starch accumulation in the leaf sheaths reached a maximum 10 to 11 weeks after transplanting, the time of development of the rice panicle. Maximal concentration of free sugars occurred earlier. Starch and sugars in the leaf sheaths and culm decreased rapidly during grain development.During starch accumulation, the starch granules of the leaf sheaths increased slightly in size and its gelatinization temperature decreased. The molecular size of amylose and amylopectin and amylose content of the starch were similar in both culm and leaf sheaths.Changes in the level of soluble protein paralleled changes in starch level in the leaf sheaths. Among the enzymes, only synthetase bound to the starch granule paralleled the level of starch in the leaf sheaths and in the culm. ADP-glucose, but not UDP-glucose, was utilized as a glucosyl donor by these starch synthetases. Zymograms of these extracts showed only one alpha-amylase band, one beta-amylase band, two phosphorylase bands, and one Q-enzyme band.

Effect of saline soil on grain quality of rices differing in salinity tolerance.

Four varieties of rice, differing in salinity tolerance and grown in saline soil (electrical conductivity 5-6 dS/m) at Sadhoke, Punjab, Pakistan, had lighter grain and higher Na content than control samples. Grains of three out of the four rices grown on saline soils had higher brown rice protein (higher nutritional value), less translucent grain, lower starch and amylose content, and lower K than their control samples, but these differences were not related to salinity tolerance. Alkali spreading value and gel consistency were not affected by culture in saline soil. Cooked rice Instron hardness increased in saline culture in two higher-protein samples of the four rices. Amylograph peak viscosity was suppressed by saline culture.