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a Dep. of Plant Sciences and Plant Pathology, Montana State Univ., Bozeman, MT 59717-3140
b Northern Agric. Res. Center, Star Route No. 36, Box 43, Havre, MT 59501
c Central Agric. Res. Center, HC90-Box 20, Moccasin, MT 59462
d Western Triangle Agric. Res. Center, P.O. Box 974, Conrad, MT 59425
e Southern Agric. Res. Center, 748 Railroad Highway, Huntley, MT 59037
* Corresponding author (bruckner{at}montana.edu)
| ABSTRACT |
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Abbreviations: AFLP, amplification fragment length polymorphism FSV, flour swelling volume HRW, hard red winter L-DOPA, L-3,4-dihydroxyphenyl alanine PPO, polyphenol oxidase SDSS, sodium dodecyl sulfate sedimentation
| INTRODUCTION |
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Asian noodles are a rapidly expanding end use for world wheat consumption. In major Asian wheat importing countries, 30 to 45% of the wheat consumed is used in noodles (Miskelly, 1996). A variety of noodle types are manufactured throughout Asia. Asian noodles can be classified based on ingredients, by the presence or absence of alkaline salt in the formula, by the width of the noodle strands, and by how they are processed and consumed (Hou and Kruk, 1998). Key quality attributes in the evaluation of wheat flour for noodle making include process properties, noodle color, and noodle texture. All noodle types require good brightness. Color should be white or yellow depending on the noodle type, having minimal discoloration with time (Hou and Kruk, 1998). Various noodle types also vary in hardness and texture requirements, with different noodle types requiring different protein content and dough strength.
Polyphenol oxidase in wheat flour causes reduced noodle brightness and decreased color stability across time (Baik et al., 1994a, 1995; Kruger et al., 1994). Polyphenol oxidase activity varies among wheat cultivars and is influenced by growing environment (Baik et al., 1994a; Park et al., 1997). Baik et al. (1995) reported that within a cultivar, noodle discoloration is more affected by protein content than by PPO. However, genetic differences between cultivars for noodle discoloration are due primarily to PPO (Baik et al., 1995). Windes and Souza (1995) reported that PPO was highly heritable with limited genotype x environment interaction. However, Park et al. (1997) reported that variability in PPO activity associated with growing locations was greater than variability among genotypes, although both sources of variation and their interaction were significant. Hatcher and Kruger (1993) demonstrated that PPO activity is closely associated with the bran component of milled wheat and increases with increasing bran contamination in the millstreams. Thus, wheats with genetically low PPO activity can be milled to a higher extraction level and retain greater value in milling and noodle production.
Decreased noodle brightness and increased time-dependent noodle discoloration have also been associated with high grain protein content (Baik et al., 1994a, 1995; Lang et al., 1998; Miskelly, 1984; Oh et al., 1985). Feillet et al. (2000) reviewed research showing that higher protein content was associated with higher pasta brownness in durum wheat {Triticum durum Desf. [= T. turgidum subsp. durum (Desf.) Husn.]}. Kobrehel et al. (1974) demonstrated conclusively that durum wheat protein content was positively correlated to pasta brownness and that the relationship between protein and pasta brownness was cultivar dependent.
High-protein hard wheats with strong gluten are most suitable for noodle types requiring firmer texture and bite characteristics such as Chinese raw noodles (Hou and Kruk, 1998). Japanese white salted or "Udon" noodles are softer in texture requiring lower protein soft wheat flour with modified starch, as indicated by an association of high starch viscosity with high flour swelling volume (FSV) and a granule bound starch synthase (GBSS-4A) null genotype (Zhao et al., 1998). Generally, flour protein content is positively correlated with noodle hardness. Baik et al. (1994b) reported that both protein content and SDS sedimentation volume were highly correlated with noodle texture profile analysis traits. Crosbie et al. (1999) reported that FSV and flour pasting characteristics (peak viscosity and breakdown) were negatively correlated with total texture score of Japanese alkaline noodles (ramen), while protein quality, as indicated by farinograph stability, was positively correlated with total texture score. Ross et al. (1997) showed that FSV was negatively correlated with alkaline noodle firmness and elasticity, and that flour protein content and SDS sedimentation volumes were significantly associated with noodle firmness. Lang et al. (1998) demonstrated positive correlations between raw Chinese noodle hardness and flour protein in two of three high protein Montana environments.
Lang et al. (1998) demonstrated the potential difficulty in developing high protein hard wheats for semiarid environments that are suitable for both bread and Asian noodle products. The primary challenge is to improve noodle color without negatively impacting loaf volume and other bread quality traits. Since we are interested in developing cultivars with superior Asian noodle qualities under high protein conditions, we would like to determine if negative correlations between bread and noodle quality traits are due to specific genes with contrasting effects on bread and noodle quality, or are caused by nongenetic factors. Blake et al. (1999), in a set of 31 hard white spring wheat lines, found 28 amplification fragment length polymorphism (AFLP) markers that were significantly associated with bread or raw Chinese noodle traits, including AFLP markers showing noodle color negatively related to flour protein and loaf volume.
The experiments described here reflect research designed to assess levels of environmental and genotypic variation for various Asian-style noodle color and texture characteristics as well as bread quality and to determine the appropriate combination of genetic traits necessary for cultivars targeted toward both bread and Asian noodle products. Specifically, this study evaluates raw Chinese noodle and bread quality of 16 adapted winter wheat genotypes grown in eight Montana environments. Responses and relationships within individual cultivars were examined in a subset of nine cultivars grown in 12 environments. Additionally, association among protein content, SDS sedimentation volume, and PPO is examined in two large nonselected, early-generation germplasm sets.
| MATERIALS AND METHODS |
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68% of the state's 2000 winter wheat acreage, Neeley (CItr 17860), Rampart (PI 593889), Tiber (PI517194), Morgan (PI 599336), Rocky (CItr 17879), and Redwin (CItr 17844) (Stringer, 2001). Genotypes also included additional cultivars and advanced breeding lines adapted to Montana: Akron (PI 584504), BigSky (PI 619166, NuWest/Tiber), Erhardt (PI 564761), McGuire (PI 593890), Niobrara (PI 584996), NuWest (PI 586806), Promontory (PI 555458), NuSky (PI 619167, NuWest/Tiber), MT9514 (MT8030/Norstar), and MT9524 (NuWest/Tiber). All genotypes were hard red winter (HRW) wheats except hard white winter wheats NuWest and NuSky. Relative PPO level determined across multiple environments [L-DOPA (L-3,4-dihydroxyphenyl alanine) assay] was high or very high in all genotypes except Promontory (very low), NuWest and NuSky (low), and Redwin (medium). Nine of the same genotypes (BigSky, McGuire, Neeley, NuWest, Promontory, Rampart, Rocky, Tiber, and NuSky) were also grown at Bozeman, Havre, Moccasin, and Huntley, MT, in 1999. Experimental entries were part of larger 49-entry yield trials planted in a lattice design with three replications in each environment. A single bulked sample of grain for each environment-genotype combination was used for quality analysis.
Associations among early-generation quality parameters were evaluated in large, diverse sets of headrow selections made at Bozeman, MT, in 1998 and 1999. The 1998 early-generation germplasm set included 1465 F5-derived F6 selected headrows and 105 harvested check rows of Judith (PI 584526) and Rampart HRW wheat. The 1999 early-generation germplasm set included 1580 F5-derived F6 selected headrows and 158 harvested check rows of Judith, Rampart, and Erhardt HRW wheat. In each year, selections and check rows were harvested from
24 000 headrows, grouped by population, with a reference cultivar set incorporated at 50-row increments. All selections were made visually based on agronomic characteristics with no end-use quality information other than pedigree.
Quality Analyses
Grain harvested from all yield trials was analyzed for milling and bread-baking properties using approved methods [American Association of Cereal Chemists (AACC), 1995]. Protein content of whole grain was measured with a Tecator Infratec 1225 Grain Analyzer (Foss North America, Silver Spring, MD). Flour protein was determined with a Technicon InfraAlyzer 400 NIR system (Technicon Industrial Systems, Tarrytown, NY) using the combustion method as the reference method. Wheat was milled on a Brabender Automat Mill (South Hackensack, NJ) after a two-stage temper to 15% moisture. Mixogram dough properties were evaluated using AACC method 54-40. Standard bake tests were conducted using AACC method 10-10B. Color and texture of noodles was determined using the same flours used in the bread analyses. Additionally, a sample of Australian Standard White wheat (mixture of Australian-grown white wheat cultivars obtained from U.S. Wheat Associates, Portland, OR) was used as a laboratory standard. Chinese raw noodles were prepared from 100 g flour and 29.2 mL salt water solution (4.29% w/v) added to the flour during a 30-s time period. Mixing continued on a National pin mixer (National Mfg., Lincoln, NE) for 5 min. After mixing, dough was rested in a plastic bag for 30 min at room temperature, then folded and passed through an Ohtake Laboratory noodle machine (Ohtake Manufacturing Co., Ltd., Tokyo) three times at a gap size of 5 mm and a temperature of 30°C. After a second 30-min rest at room temperature, dough was successively passed through five progressively narrower roller settings to achieve a final noodle sheet thickness of 1.2 mm. Dough sheets were stored in plastic bags at room temperature and evaluated for color using the Minolta CR-310 Chroma Meter (Minolta, Ramsey, NJ) using the Commission Internationale De l'Eclairage (CIE) L* (brightness) a* (red-green) b* (yellow-blue) color system 0 and 24 h after sheeting. Texture profile analysis (springiness, cohesiveness, adhesiveness, hardness, chewiness) was performed on five strands of rinsed noodles 0 and 5 min after cooking using a TA-XT2 Texture Analyzer (Texture Technologies Corp., Scarsdale, NY) assembled with a 6-mm flat lexan probe. Scale of noodle texture parameters changed between 1997 and 1998 due to a change in noodle width from a No. 10 to a No. 12 noodle slitter (2.5-mm noodle width). Mean hardness (0 min) in 1997 was 525 g compared with mean noodle hardness (0 min) in 1998 of 1422 g.
Noodle scores were calculated for each sample using combined criteria of color stability and subjective evaluations of noodle processing properties. Noodle scores are additive, incorporating categories for machining properties (20 points), dough sheet appearance (5 points), and color stability (30 points). Experimental samples were scored (110) against the standard control flour in each category. The standard control flour (Australian Standard White) was assigned a score of 7 in each category (noodle score = 385).
Protein content of early-generation selections was determined using the Tecator InfraTec 1225 Grain Analyzer. For SDS sedimentation and PPO evaluation, samples were ground in an UDY cyclone sample mill (UDY Corp., Fort Collins, CO) with a 0.5-mm screen. SDS sedimentation was determined using methods modified from Axford et al. (1979) and Preston et al. (1982). SDS-lactic acid reagent was prepared by dissolving 20 g of SDS in 1 L of water and adding 20 mL of stock diluted lactic acid solution (one part lactic acid plus eight parts water by volume). Briefly, a 0.45-g whole meal sample was placed in a 10-mL graduated cylinder, 5 mL water containing methylene blue was added, and cylinders were vortexed at 0, 2, and 4 min. Immediately after the last vortex, 5 mL SDS-lactic acid reagent was added and cylinders were mixed four times by gentle inversion using an aliquot mixer, with inversion repeated at 2, 4, and 6 min. After 20 min of settle time, sedimentation volume was determined. Polyphenol oxidase was determined using a modification of the L-DOPA assay reported by Anderson and Morris (2001). Approximately 300 mg of UDY-ground wholemeal flour of each sample was placed in a single well of a 12-well microtiter plate. Durum wheat meal was used as a standard in each microtiter plate. A 1.5-mL aliquot of 5 mM L-DOPA in 50 mM MOPS (3-[N-morpholino] propanesulfonic acid) solution, pH 6.5, was pipetted into each well and stirred. Microtiter plates were rotated for 30 min on a reciprocating shaker at 150 rpm. Samples were visually rated on a 0 to 5 scale (0 = lightest, 5 = darkest; durum check = 0).
Statistical Analyses
For quality traits, analyses of variance were conducted across eight environments. Significance of mean genotype differences were determined using the environment x genotype mean square as an error term. Genotypes were subjectively classified into poor, acceptable, good, and outstanding groups for noodle and bread quality traits based on relative mean response and mean separation by LSD.
Associations among bread and noodle quality traits were evaluated in eight environments using Pearson correlation coefficients and pooled correlation coefficients calculated across environments following chi-square analyses to test for homogeneity of correlations. Correlation coefficients among bread and Asian noodle quality traits were also calculated for nine individual cultivars and pooled across cultivars using the 12-environment data set. Relationships among flour protein and initial and 24-h noodle brightness for individual cultivars were characterized by regression analysis. Heterogeneity of regression coefficients among cultivars was tested using procedures outlined in Steel et al. (1997).
Associations among grain protein content, PPO level, and SDSS volume in early-generation germplasm sets were compared by correlation analysis for individual years, and in subsets of progeny derived from populations segregating for PPO level. In addition, correlation analyses were conducted in each of 17 individual wheat populations segregating for PPO level (minimum of 10 progeny). Populations were judged to be segregating for PPO level when multiple progeny exhibited at least a two-class difference in PPO level. Homogeneity of correlation coefficients in the 17 populations segregating for PPO level was evaluated using chi-square tests and pooled correlation coefficients calculated across the populations.
| RESULTS |
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Based on correlation analyses of genotypes in and across environments, genotypes with higher flour protein produced noodles with lower initial and 24-h brightness (L*) and lower noodle scores (Table 3) . The negative correlation between flour protein and noodle brightness was consistent in all production environments, and of similar magnitude to the positive correlation of flour protein with bake absorption (pooled r = 0.75, p < 0.01) and loaf volume (pooled r = 0.79, p < 0.01). Loaf volume, but not bake absorption, was negatively correlated with initial and 24-h noodle brightness (L*). Flour protein was positively correlated with noodle hardness in only two of the eight production environments (Table 3). Bake absorption was positively correlated with noodle hardness in three of the eight production environments. Loaf volume was not correlated with noodle hardness in pooled analyses. Initial and 24-h noodle yellowness (b*) was not correlated with flour protein (data not shown).
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Nonselected Germplasm
Two large and diverse early-generation sets of headrow bulks selected with no quality information were screened for grain protein content, SDSS volume, and relative PPO level. Mean grain protein content and SDSS volume were similar in all groups classified by PPO level in both years (Table 5)
. Correlation of grain protein with PPO level across all populations was low but significant in both 1998 and 1999 (1998, r = 0.073, P < 0.05, n = 1465; 1999, r = 0.072, P < 0.05, n = 1580). In subsets of germplasm segregating for PPO, correlations of grain protein with PPO were nonsignificant (1998, r = -0.043, n = 278, 31 populations; 1999, r = -0.072, n = 193, 23 populations). Correlation of PPO level with SDSS volume across all populations was nonsignificant in both years (1998, r = -0.042; 1999, r = 0.010). In subsets of germplasm from populations segregating for PPO level correlations of PPO level and SDSS volume were also nonsignificant (1998, r = -0.121, n = 278, 31 populations; 1999, r = 0.094, n = 193, 23 populations). Correlation analyses were also conducted in each of the 17 largest individual wheat populations segregating for PPO level (minimum of 10 progeny). A significant correlation between grain protein and PPO level occurred in only one of 17 populations (Table 6)
. Correlation coefficients for grain protein and PPO were homogeneous across populations with a pooled correlation coefficient of zero. Correlation coefficients of PPO and SDSS volume were homogeneous and nonsignificant in all 17 populations segregating for PPO (pooled r = 0.07).
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| DISCUSSION |
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This set of elite U.S. germplasm was developed and released before substantial efforts to improve Asian noodle quality were made. Thus selection was primarily for protein content and quality and for dough mixing and bread baking characteristics. Despite this, analysis of bread and raw Chinese noodle qualities in this germplasm (Table 2) indicates we can and have developed cultivars suitable for dual-purpose utilization for both bread and Asian noodle products. Three low-PPO cultivars, NuSky, NuWest, and Promontory, have relatively good raw Chinese noodle and bread baking qualities. Although acceptable for bread products, these cultivars are of intermediate protein level and in general are lower in absorption and loaf volume than the best bread quality cultivars. Several high-PPO genotypes (Rocky, MT9524, BigSky, Neeley) were also identified that had high initial noodle brightness and good noodle color stability but average bread baking characteristics. Rocky and Neeley have been dominant cultivars in Montana for more than a decade. Several high protein cultivars (Redwin, Erhardt, Rampart, and McGuire) had excellent bread baking characteristics but very poor noodle color characteristics.
The moisture-limited environments of Montana and other areas of the U.S. northern Great Plains are ideal for production of high protein hard wheat. Our correlation results indicate that environmental conditions favoring development of high wheat protein and superior bread quality detrimentally affected noodle brightness and color stability. Environmentally-induced variation in flour protein content was not associated with noodle hardness (Table 4). Genotypes producing the hardest Chinese noodles included high protein cultivars (McGuire, Rampart) as well as intermediate protein lines (NuSky, MT9524, BigSky).
Wheat breeding programs of the northern U.S. Great Plains and Pacific Northwest are interested in developing wheats with enhanced quality and value which fit into existing environmental constraints and cropping systems. In moisture-limited environments, production of high protein wheats is a priority. Our strategy is to develop genotypes with improved Asian noodle characteristics within high protein backgrounds rather than modify management systems to produce lower protein noodle wheats. For dual-purpose utilization in both bread and Asian noodle products, we hypothesize that a high protein, low-PPO genotype with good protein quality would be appropriate. Such a wheat cultivar was not available for testing in the current study.
As pointed out by Lang et al. (1998), the negative correlations among bread and noodle color traits do not preclude successful improvement of one trait without negatively impacting the correlated trait. A pertinent example is grain yield and grain protein content, which are negatively correlated in wheat. Through successful selection, grain yield has been increased while maintaining protein content at acceptable levels (Lanning et al., 1994; Sears et al., 1991). Our finding of little or no association between grain protein content, SDS sedimentation volume, and level of PPO in two large and diverse sets of early-generation selections is noteworthy. In specific populations segregating for PPO, grain protein and SDSS volume were not correlated with PPO level. These relationships in unselected populations indicate that successful selection could combine high protein content and high protein quality with low PPO levels. Such genotypes have potential to combine superior bread quality with superior Asian noodle qualities. On the basis of existing germplasm and available selection technology, such an objective appears attainable.
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Received for publication June 22, 2001.
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