Quality protein maize: a biochemical study of enzymes involved in lysine metabolism.

Quality protein maize (QPM) varieties have been produced by the introduction of opaque-2 modifier genes. Two QPM varieties, BR451 and BR473, a wild type and an opaque-2 variety, have been used to study key enzymes controlling lysine metabolism in the endosperm during development. Aspartate kinase and homoserine dehydrogenase enzymes, which are involved in lysine and threonine biosynthesis, respectively, exhibited identical activity patterns during endosperm development, with a maximum specific activity at 16 days after pollination. The QPM varieties exhibited higher levels of aspartate kinase activity in the endosperm, suggesting an increased rate of lysine biosynthesis when compared to the opaque-2 and wild-type genotypes. Similar results were observed for the lysine ketoglutarate reductase and saccharopine dehydrogenase enzymes, which form a single bifunctional polypetide involved in endosperm lysine degradation. Both enzyme activities were strongly reduced in the opaque-2 maize variety when compared to the wild-type maize, whereas the QPM varieties exhibited even lower levels of lysine ketoglutarate reductase-saccharopine dehydrogenase activities when compared to the opaque-2 variety. The developmental pattern of enzyme activity showed a different profile when compared to the enzymes involved in lysine biosynthesis, with activity being detected only 12-16 days after pollination (DAP) and maximum activities approximately 24 DAP. These results also suggest that the modifier genes have intensified the effect of the opaque-2 mutation on lysine ketoglutarate reductase-saccharopine dehydrogenase. These alterations lead to an increase in soluble lysine in the endosperm of the QPM varieties when compared to the opaque-2 and wild type.     

Near-infrared reflectance spectroscopy (NIRS) for protein, tryptophan, and lysine evaluation in quality protein maize (QPM) breeding programs

Quality protein maize (QPM) has approximately twice the tryptophan (Trp) and lysine (Lys) concentrations in protein compared to normal maize. Because several genetic systems control the protein quality of QPM, it is essential to regularly monitor Trp and/or Lys in breeding programs. Our objective was to examine the potential of near-infrared reflectance spectroscopy (NIRS) to enhance the efficiency of QPM research efforts by partially replacing more expensive and time-consuming wet chemistry analysis. More than 276 maize samples were used to develop NIRS models for protein content (PC), Trp, and Lys. The standard error of prediction (SEP) for the calibration and the coefficient of determination for validation (R(2)(v)) were 0.26 and 0.96 for PC, 0.005 and 0.85 for Trp, and 0.02 and 0.75 for Lys. When the NIRS models were used to evaluate 266 S2 lines from five QPM breeding populations, the coefficients of determination between NIRS and the chemical data were 0.94, 0.76, and 0.80 for PC, Trp, and Lys, respectively. Therefore, the NIRS models can be used to support the QPM breeding efforts.     

Maize Endosperm Proteins That Contribute to Endosperm Lysine Content

Nonzein proteins were analyzed using SDS-PAGE to study their relationship to lysine level in mature endosperm of three isogenic lines and 29 inbred lines. The majority of nonzein proteins are glutelins that contain a relatively high lysine content. Trinitrobenzene-sulfonic acid was used to visualize high-lysine proteins on nitrocellulose membrane. Seven intense yellow bands (high-lysine proteins) were found in the glutelin fraction: 35, 43, 48, 52, 84, 92, and 100 kDa. These seven glutelin proteins were also significantly correlated to each other. Lysine content of the triads correlated with three glutelin proteins (35, 43, and 52 kDa), while lysine content of 29 inbred lines correlated with six glutelin proteins (all except 52-kDa protein). Immunoassays based on these higher lysine glutelin proteins provide an effective screening method in the presence of variable zein levels. Retention or improvement of lysine content is possible using this information in a maize improvement program.     

优质蛋白玉米籽粒品质性状组间典型相关性研究

研究优质蛋白玉米籽粒品质性状组间典型相关性结果表明,2个半硬质胚乳奥帕克-2玉米群体(“中群14QPMC3”和“陕综5号QPMC0”)籽粒物理性状中硬质度、籽粒密度、百粒重与胚比、百粒体积呈负相关效应,营养品质中赖氨酸含量、蛋白质品质性状为其核心,籽粒密度(或硬质度)可作为蛋白质品质的间接选择指标;籽粒物理性状、蛋白质品质性状在赖氨酸产量构成中起决定性作用,全籽粒蛋白质品质与胚乳蛋白质品质紧密相关,而与胚蛋白质品质无关,高赖氨酸高油分育种相统一。     

Production and Nutritional Evaluation of Liquefied Weaning Foods from Malted Sorghum,Quality Protein Maize,and Other Cereals

Different concentrations of sorghum diastatic malt (SDM) were added to pregelatinized pastes from regular maize flour with the aim of hydrolyzing the starch to produce liquefied foods with 15% solids. Viscosities of the blends decreased as the concentration of SDM increased. Addition of 6.66% SDM based on total amount of solids reduced viscosity by ≈50% when compared with a food that did not contain any SDM. Addition of 33.3 or 46.6% SDM reduced viscosity by ≈70 or 75%, respectively. Most of the reduction in viscosity occurred within 1–3 min of incubation with warm water. Weanling rats were fed a combination 33.3% SDM and 66.6% of either quality protein maize (QPM), regular maize (RMZ) or decorticated pearl millet (DPM) to estimate protein efficiency ratios (PER), protein digestibility, biological value (BV), and net protein utilization (NPU). Rat growth was positively correlated with dietary lysine content and essential amino acid (EAA) scores; therefore, animals fed QPM weanling food had significantly higher (P < 0.05) protein digestibility corrected EAA scores, PER, BV, and NPU than counterparts fed diets based on RMZ or DPM. This demonstrates that it is feasible to produce nutritious liquefied weaning foods blending 33.3% SDM with 66.6% QPM using simple processing techniques.     

Technological and Nutritional Properties of Flours and Tortillas from Nixtamalized and Extruded Quality Protein Maize (Zea mays L.)

Nixtamalized and extruded flours from quality protein maize (QPM, V‐537C) and tortillas made from them were evaluated for some technological and nutritional properties and compared with the commercial brand MASECA. Both QPM flours showed higher (P < 0.05) protein content, total color difference, pH, available lysine, and lower (P < 0.05) total starch content, Hunter L value, water absorption index, gelatinization enthalpy, resistant starch, and retrograded resistant starch than nixtamalized MASECA flour. Tortillas from nixtamalized and extruded QPM flours had higher contents of essential amino acids than tortillas from MASECA flour, except for leucine. Tortillas from processed QPM flours also showed higher (P < 0.05) values of the nutritional indicators calculated protein efficiency ratio (C‐PER 1.80–1.85 vs. 1.04), apparent and true in vivo protein digestibility (78.4‐79.1 vs. 75.6% and 76.4–77.4 vs. 74.2%, respectively), PER (2.30–2.43 vs. 1.31), net protein retention (NPR; 2.88–2.89 vs. 2.11), and protein digestibility corrected amino acid score (PDCAAS; 54–55 vs. 29% based on preschool children and 100 vs. 85% based on adults) than MASECA flour. The use of QPM for flour and tortilla preparation may have a positive effect on the nutritional status of people from countries where these products are widely consumed.     

Assessment of the protein quality of nine northern adapted yellow and brown seed coated soybean cultivars by amino acid analysis

Accurate and detailed amino acid determinations were carried out on nine northern adapted soybean cultivars to ascertain whether their amino acid profiles could be used as potentially useful indices for assessing their protein quality. The cultivars were Maple Amber, Maple Donovan, Maple Glen, Maple Isle, Maple Presto, Maple Ridge, and three brown seed coat near-isogenic lines, Maple Presto Brown, Maple Ridge Brown, and Maple Arrow Brown. Their total protein and amino acid composition were compared with those of an established cultivar, Maple Arrow. Mean protein values for the new cultivars ranged from 30.1 to 33.1% compared to Maple Arrow, which was 33.2%. The total nitrogen content was also variable among these cultivars, ranging from 5.0 to 5.4%. All nine Maple series soybean cultivars were higher in their essential amino acid (EAA) content, that is, EAA(9) = 45.2-46.5%, than the FAO/WHO reference protein pattern value of EAA(9) = 33.9%, for a 2-5-year-old child. Each of the nine new soybean cultivars was limited only in methionine and to a lesser extent in valine and isoleucine and had a protein digestibility corrected amino acid score of 91% for all cultivars, compared to the value of egg protein (97%). These results suggest that the most accurate evaluation of protein quality in soybeans, and possibly other legumes and cereals, is by the protein digestibility-corrected amino acid score.     

Biological nitrogen fixation in Azospirillum strain-maize genotype associations as evaluated by the 15N isotope dilution technique

Few studies of the inoculation of cereal crops with N₂-fixing bacteria have included more than one or two plant genotypes. In a recent study performed in Argentina using 12 different maize genotypes, it was found in 2 consecutive field experiments that several of them responded consistently, either negatively or positively, to inoculation with a mixture of strains of Azospirillum spp. The present study in post was performed to investigate the effect of inoculation of individual strains (and a mixture) of Azospirillum spp., and their nitrate reductase negative (NR-) mutants, on the growth of four of these maize genotypes. Two of these genotypes were grown in ¹⁵N-labelled soil with the aim of quantifying any contributions of biological N₂ fixation. Two genotypes (Morgan 318 and Dekalb 4D-70) produced similar increases in grain yield when they were inoculated with a mixture of Azospirillum spp. strains or fertilized with the equivalent of 100 kg N ha^-1. The other genotypes (Dekalb 2F-11 and CMS 22) showed little response to inoculation or N fertilization. The Morgan 318 and Dekalb 4D-70 genotypes showed a large increase in total N accumulation, suggesting that the response was due to increased N acquisition, but not due to bacterial nitrate reductase as the NR- mutants generally caused plant responses similar to those of the parent strains. Despite problems with the stabilization of the ¹⁵N enrichment in the soil, the ¹⁵N isotope dilution results indicated that there were very significant biological nitrogen fixation (BNF) contributions to the Dekalb 4D-70 and CMS 22 maize genotypes.Dedicated to Professor J.C.G. Ottow on the occasion of his 60th birthday     

Improving nutritional quality of maize proteins by expressing sense and antisense zein genes

The predominant proteins in maize grain are a family of alcohol-soluble prolamin storage proteins called zeins. They account for >50% of total seed proteins but are deficient in several essential amino acids. As a result, the corn grain is considered to be nutritionally poor for monogastric animals with respect to key essential amino acids, most notably lysine, tryptophan, and methionine. Thus, corn mutants with reduced levels of zeins, such as opaque-2 (o2), have been demonstrated to possess grain with improved nutritional quality characteristics. The o2 mutant has a superior amino acid composition and has been used through conventional breeding to develop Quality Protein Maize (QPM) for human and animal consumption in developing countries. With the understanding of molecular genetics of zeins and progress in biotechnology, an alternative approach to zein reduction is explored here. Through the targeted reduction of the 19-kDa alpha-zeins, increased levels of lysine, tryptophan, and methionine have been engineered in grain of transgenic hybrids. Currently, the agronomic properties and nutritional values of these transgenic lines are being evaluated.     

Effect of six decades of selective breeding on soybean protein composition and quality: a biochemical and molecular analysis

To evaluate the extent of the genetic change and its effects on the seed protein composition of soybean cultivars released during the past 60 years, representative ancestral cultivars and those derived from selective breeding were grown in a side-by-side comparison. Total seed protein content, determined by combustion analysis of nitrogen, revealed a decline in the protein content after decades of selection and breeding. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis comparison of protein profiles of the soybean cultivars indicated that relative expression of most of the seed storage proteins had not varied substantially from the ancestral lines to the present commercial cultivars. There was noticeably less beta-subunit of beta-conglycinin, a protein devoid of sulfur amino acids, in the modern cultivars represented by Mustang, Pioneer 93B09, and Asgrow 3602. Comparison of the amino acid profiles of soybean seed, a benchmark of the protein's nutritional quality, revealed that the ancestral progenitor, G. soja, was significantly higher in cysteine, glutamic acid, histidine, and arginine than either the ancestral or the modern cultivars. Selective breeding over the past 60 years minimally affected the overall amino acid composition. The degree of divergence in the DNA sequence of the genes encoding glycinin and beta-conglycinin in the ancestral and modern cultivars was investigated using Southern hybridization and the polymerase chain reaction. Even though some restriction fragment polymorphisms could be detected, overall, the banding patterns were remarkably similar among the ancestral cultivars and those derived from them, suggesting a high degree of conservation of seed-storage protein genes. The results of our study suggest that selection and breeding for yield during the past 60 years had no major influence on the protein composition, ostensibly because of limited genetic diversity among the parental lines.     

Total and reactive lysine contents in selected cereal-based food products

The aim of the study was to determine and compare reactive and total lysine contents in a range of breakfast cereal products. Crude fiber, fat, ash, and crude protein contents of 20 breakfast cereal products ranged from 4 to 38, 14 to 144, 7 to 32, and 52 to 253 g/kg, respectively. The concentrations of glutamic acid (18.7-32.1 g/100 g protein) and proline (4.7-10.8 g/100 g protein) were high while those of the amino acids methionine (1.2-2.0 g/100 g protein) and histidine (1.2-3.3 g/100 g protein) were relatively low. There was a strong relationship between reactive lysine determined using the guanidination and fluorodinitrobenzene methods (R = 0.99). The total lysine content, determined after conventional acid hydrolysis, ranged from 0.8 to 3.7 g/100 g protein, while the reactive lysine content (guanidination) ranged from 0.4 to 2.8 g/100 g protein. Reactive lysine was 20-54% lower than total lysine in the cereal products. The large differences between total and reactive lysine suggest a considerable loss of lysine in the breakfast cereals tested.     

氮素对不同类型玉米蛋白质及其组分和相关酶活性的影响

以普通玉米掖单22和高油含量玉米高油115为材料,测定了不同供氮水平下子粒蛋白质含量及其组分以及叶片和子粒中硝酸还原酶(NRase)、谷氨酰胺合成酶(GS)和谷氨酸脱氢酶(GDH)活性的品种差异。结果表明,玉米子粒蛋白质及其组分含量,均为高油115显著高于掖单22,其中醇溶蛋白和白蛋白含量增幅明显,且随施氮量的增加而提高。不同供氮条件下,两种类型玉米叶片中NRase、GS活性和子粒中GS、GDH活性的动态变化一致,NRase活性自灌浆初期至成熟期一直下降,GS、GDH活性呈单峰曲线,在授粉后20～30d达到高峰;增施氮肥有利于酶的活性维持较高水平。掖单22叶片中NRase活性高于高油115;叶片GS和子粒GDH活性显著低于高油115,这可能是其蛋白质含量较低的生理原因之一。     

Effects of thermal processing and storage on available lysine and furfural compounds contents of infant formulas

The Maillard reaction-related effects that thermal treatments during the manufacturing process and storage (at 20 and 37 degrees C) have on powdered adapted and follow-up milk-based infant formulas were estimated by measuring the available lysine and furfural compounds contents of raw cow milk used in manufacturing, intermediate products and formulas. A fluorimetric method was used to measure the available lysine contents, and free and total furfural compounds were determined by HPLC. Statistically significant losses in available lysine (about 20%) in the infant formulas with respect to raw milk were found. The storage period did not affect the available lysine contents of adapted formulas but reduced (16%) the contents of the follow-up ones (from 6.61 to 5.33 g/100 g of protein). No furfural compounds were detected in raw milk, and free and total furyl methyl ketone (FMC) and methylfurfural (MF) were not observed in the analyzed samples. After 6 months of storage, an increase in free hydroxymethylfurfural (HMF) (from 0.34 to 0.77 mg/100 g of protein) and furfural (F) (from nondetectable to 0.1 mg/100 g of protein) in adapted formulas and free HMF (from 1.84 to 2.62 mg/100 g of protein) in follow-up formulas was observed.     

Yield and protein quality of five soft red winter wheat cultivars as affected by nitrogen fertilization rate

Abstract

This study reports the relationship between N levels (22, 56 and 112 kg/ha) and its effect on yield and protein quality for five cultivars of soft winter wheat (Triticum aestivum L. em. Thell). The objective was to determine the relationship of amino acid content and total protein at various levels of N. The 56 and 112 kg N/ha rates resulted in higher wheat yields than the 22 kg N rate over the 3 yr. mean. A cultivar x N level interaction for yield occurred when certain cultivars did not respond to higher N rates in 1972. Total seed protein increased with each increase in If. Increasing N levels generally resulted in a higher amlno acid content of the seed. Cultivar effects were found for 6 of the 17 amino acids (threonine, serine, glutamic acid, proline, glycine and lysine) indicating that cultivars differed in total amino acid content. Cultivars Arthur and Holley gave the greatest response (percentage amino acid) to N application. Coker 68–8 was intermediate in response while Coker 68–15 and Blueboy showed small inconsistent responses to H application.     

Invertase activity limits grain yield of maize under salt stress

Salt stress reduces grain yield of maize (Zea mays L.) due to poor kernel setting but not due to decreased grain filling. In the present study, it was tested whether acid invertase activity is decreased in developing kernels of maize under salt stress, and if assimilate supply is limited. The relatively salt‐sensitive maize hybrid Pioneer 3906 was compared with the more salt‐resistant hybrid SR 12. Salt stress caused a significant decrease in grain yield which was due to a 50% decrease in kernel number. No source limitation was observed, as the sucrose concentrations in kernels were significantly increased under salt stress for both genotypes. In contrast, glucose and fructose concentrations in kernels were significantly decreased. Salt stress caused a significant inhibition of soluble acid invertase activity to 19% in hydroponics 5 d after pollination (5 DAP) and to 50% in the soil culture experiment (2 DAP). The decrease in enzyme activity was the same for both genotypes. In the soil experiment, the highest soluble acid invertase activity was found 2 DAP with a steep decline until 8 DAP in Pioneer 3906. It is concluded that a decrease in acid invertase activity is a key factor associated with limited kernel setting under salt stress but additional factors may be responsible for genotypic differences.     

干旱胁迫下施钾水平对油菜生长特性、籽粒品质和钾素利用的影响

[目的]探究长期干旱胁迫下不同施钾水平对油菜生长、籽粒品质、钾素利用的影响,旨在明确不同钾肥水平下油菜通过调节生长和营养分配应对干旱胁迫的机制,为油菜抗旱栽培提供科学依据.[方法]以抗旱油菜品种油研57和干旱敏感品种川油36为试验材料,采用盆栽土培试验,每盆装风干土10 kg,设置K2O施用量0、80和160 mg/k...     

Variability of oil content and of major fatty acid composition in almond (Prunus amygdalus Batsch) and its relationship with kernel quality

Oil content and fatty acid composition were determined for two years in the kernel oil of eight cultivars and 47 advanced self-compatible almond genotypes developed in an almond breeding program. Considerable variation between genotypes was found for all parameters. Oil content ranged from 48% to 67% of the total kernel dry weight but was consistent over the two years. Fatty acid composition was also very variable, with significant differences between genotypes, even in genotypes of the same progeny. Oleic acid, ranging from 63% to 78%, and linoleic acid, ranging from 12% to 27%, were the major fatty acids, showing higher values in some selections than in their parents. The large variability observed for all fatty acids and the presence of selections with higher oil and fatty acid contents than the commercial cultivars represents a very promising base to obtain new almond cultivars with oil of higher quality, satisfying the industrial and consumer sectors.     

Assessment of indigenous Nepalese soybean as a potential germplasm resource for improvement of protein in North American cultivars

Soybeans contain approximately 40% protein and 20% oil and represents an important source of protein in animal rations and human diets. Attempts are being made to increase further the overall protein content of soybeans by utilization of exotic germplasms. In this study, soybean cultivars from Nepal have been characterized and their potential as a germplasm resource for improvement of the protein content and quality of North American cultivars assessed. Soybean cultivars 'Sathia', 'Seti', 'Kavre', and 'Soida Chiny', indigenous to various regions of Nepal, contained 42-45% protein, which is significantly higher in comparison to that of the North American cultivar 'Williams 82' (39%). Fractionation of seed protein by high-resolution two-dimensional gel electrophoresis revealed differences in the protein profiles of these cultivars. Various isoelectric forms of glycinin and beta-conglycinin were identified by comparing the matrix-assisted laser desorption ionization time-of-flight mass fingerprinting data against the National Center for Biotechnology Information nonredundant database. Nepalese cultivar Sathia was distinct, lacking some isoelectric forms of acidic and basic glycinin subunits while expressing other unique forms. The contribution of these unique protein spots present in either Sathia or Williams 82 to the total protein content was quantified using scanning laser densitometry. Distinct restriction fragment length polymorphisms (RFLP) for group 1 glycinin genes were observed among the tested Nepalese genotypes, indicating sequence variation among the cultivars. Conversely, evaluation of RFLP for the genes encoding group 2 glycinins, beta-conglycinin, and Bowman-Birk proteinase inhibitors indicated a high degree of conservation in these genes. Determination of amino acid composition, a reflection of protein quality, indicated that the arginine content of the Nepalese soybeans ranged from 7.7 to 8.1%, which was 5-10% higher than the 7.4% expressed in Williams 82. Additionally, Karve and Seti contained significantly more cysteine than Williams 82. Nepalese high-protein soybeans having a desirable amino acid composition hold potential to increase the protein quality and diversity of North American cultivars.     

猪粪沼液施用对土壤氨挥发及玉米产量和品质的影响

猪粪沼液含有大量的养分,如氮、磷、钾、氨基酸、腐植酸等,但如果不及时施用,也会带来水体富营养化问题。沼液替代化肥是目前最有效的利用方式。采用田间试验方法,在春、夏两季玉米上,研究了猪粪沼液不同用量与不同运筹对土壤氨挥发损失以及玉米产量、品质等的影响。结果表明,夏玉米基施猪粪沼液氨挥发量大于春玉米,沼液的氨挥发量明显大于化肥,且沼液施用量越大,氨挥发量越大;由氨挥发导致的氮素损失量占施肥量的0.17%~2.16%,平均为0.88%。在春玉米中,单独施用化肥以及化肥与沼液各50%配施处理的产量分别为901.55 kg.667m?2和892.71 kg.667m?2,显著高于其他处理;纯化肥处理可溶性糖为103.69 g.kg?1,显著低于其他处理;纯化肥处理皮渣率为9.87%,显著高于其他处理;其他品质指标各处理间无显著差异。在夏玉米中,总体产量不如春玉米,纯化肥处理产量为523.47 kg,显著高于其他处理,化肥与沼液各50%配施处理的产量仅次于纯化肥处理,为513.41 kg。纯化肥处理的玉米品质总体上高于纯沼液处理,但沼液和化肥各50%处理玉米品质与纯化肥处理无明显差异。可见,对玉米植物而言,以50%沼液替代化学肥料,在技术上可行,可以获得与化肥处理相同的玉米产量与品质。     

Apoplast acidification is not a necessary determinant for the resistance of maize in the first phase of salt stress

In previous studies, a relation between plant growth during the first phase of salt stress and cell‐wall acidification was shown for differently resistant maize genotypes. In the present study, plants of the salt‐sensitive maize (Zea mays L.) cv. Pioneer 3906 and the salt‐resistant genotype SR 12, grown under 100 mM NaCl, showed a similar decrease in plasmalemma H⁺‐ATPase activity, while SR 12 showed less growth reduction than Pioneer 3906. From this it is concluded that maintenance of apoplast acidification is not necessary for better plant growth during the first phase of salt stress.