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It is often claimed that modern plant breeding erodes genetic diversity. Fjellheim and Rognli (2081–2086) investigation used AFLP markers to study genetic diversity within and between Nordic cultivars of the forage grass meadow fescue (Festuca pratensis Huds.) released in the period from 1917 to 2001. The genetic diversity within newly released cultivars was as large as within old cultivars, indicating that breeding has not eroded genetic diversity over time. Exchanges of breeding materials between the Nordic countries have counteracted genetic erosion and genetic differentiation between countries. The applicability of AFLP markers in cultivar identification in meadow fescue is discussed.
Effects of Irrigation and Fertilizer Rate on Wheat
The interplay between crop irrigation amount and fertilizer rate effects on hard spring wheat (Triticum aestivum L.) bread and noodle quality was evaluated in this study by Guttieri et al. (2049–2059). Reducing the amount of irrigation elevated grain protein concentration; however, it also reduced milling yield. Increased nitrogen fertilizer increased both peak flour pasting viscosity and final flour pasting viscosity. In this study, it was preferable to increase grain protein concentration by increasing fertilization rather than by reducing irrigation.
Spring Wheat Crop Production and Fusarium Head Blight
Fusarium head blight (FHB) is of growing importance on the Canadian Prairies. The objective of this study was to identify crop production factors (CPF) that were associated with FHB development in spring wheat (Triticum aestivum L.). From 1999 to 2002, 659 fields were sampled in eastern Saskatchewan for FHB levels, together with information on the agronomic practices being used. In 2 yr, Fusarium-damaged kernels (FDK) were also determined. Findings of Fernandez et al. (1908–1916) indicated that environment was the most important factor affecting disease development. Previous application of glyphosate formulations (GF), tillage system, previously-grown crop, and cultivar susceptibility were the only CPFs that affected FHB. GF application was significantly associated with higher FHB levels every year. Percentage FDK was also higher in fields previously treated with GF. Further research is needed to determine if the association between previous GF use and FHB development was a cause–effect relationship.
Grazing Management as a Control of Vaseygrass
Grazing management is potentially a cost-effective tool for control of the weed vaseygrass (Paspalum urvillei Steud) in limpograss [Hemarthria altissima (Poir.) Stapf & Hubb.] pastures. Newman and Sollenberger (2038–2043) evaluated combinations of grazing method (continuous or rotational) and intensity (15- and 30-cm stubble height) at low or high (50 or 150 kg N ha–1) N fertilization rates. Continuous stocking reduced vaseygrass cover in limpograss pastures, as did close grazing (15 cm) associated with low N fertilization rate. The authors cautioned that extended use of continuous stocking at low-stubble heights may have negative effects on limpograss stands, thus careful management of stubble height is required to avoid invasion by other undesirable species.
Turfgrass Germination Responses to Temperature
Turfgrass is often established by sowing species seed mixtures. Larsen and Bibby (2030–2037) studied the germination response to temperature of three different turfgrass species. Final germination percentage is little affected by suboptimal germination temperatures for all three species. However, Kentucky bluegrass (Poa pratensis L.) requires a longer thermal time to germinate than red fescue (Festuca rubra L. subsp. litoralis Vasey) and particularly perennial ryegrass (Lolium perenne L.). The slower germination of Kentucky bluegrass may result in a poorer competitive ability of this species during establishment and may in part explain why this species often establishes poorly when sown in seed mixtures with red fescue and perennial ryegrass.
Brazilian Maize Grain Quality
Maize (Zea mays L.) is an important crop in Brazil, and concerns about grain quality are increasingly important with increasing exports and use of grain for specific end uses. Duarte et al. (1958–1964) studied grain quality differences for a range of Brazilian maize genotypes and N application rates. Genotype had a larger impact on grain quality than did N rate. Large variation in grain yield and grain quality was found for intermediate kernel-type (semident, semiflint) genotypes grown in Brazil which presents short-term potential for producers to select hybrids that produce both high yields and good dry milling grain quality.
Germination Characteristics of Oat
Willenborg et al. (2023–2029) conducted a germination study to identify oat genotypes or seed size(s) with high germination potential under moisture stress in the northern Great Plains region. Large, medium, and small seeds of six common western Canadian oat genotypes were germinated in polyethylene glycol solutions with initial osmotic potentials ranging from 0 to –0.4 MPa at 5°C. Among genotypes, CDC Bell had the fastest median germination time (MGT) while AC Mustang had the highest final germination percentage (FGP). In all genotypes examined, decreasing seed size and osmotic potential significantly increased MGT and lowered FGP. However, these differences are generally small, and therefore the results suggest that oat germination characteristics may not be substantially improved by screening out small seed from farm-saved seed.
Forage-Type Soybean in the Southern Great Plains
The primary forage resources for livestock production in the southern Great Plains are winter wheat (Triticum aestivum L.) and warm-season grasses. Warm-season grasses are productive during late spring through summer, but quality declines with the length of growing season. Rao et al. (2068–2074) evaluated forage production patterns and nutritive values of one seed-type and three forage-type soybean [Glycine max (L.) Merr.] cultivars to determine their capacity to fill this forage gap. Forage cultivars provided high quality and quantity forage during mid-July through early-October. The forage cultivars produced 15 to 47% more leaf and 43 to 129% more stem through 120 d of growth. Forage quality was similar for all cultivars.
Breeding for Fusarium Head Blight Resistance in Spring Wheat
The development of wheat cultivars resistant to Fusarium head blight (FHB) requires screening methodologies that accurately characterize reaction to this disease. Fuentes et al. (1965–1972) evaluated 14 cultivars in FHB-screening nurseries at two locations across a 4-yr period. Most of the cultivars, including susceptible ones, expressed stability for FHB reaction. Optimum resource allocation was most affected by the number of environments, with three being the minimum to accurately characterize a genotype's resistance level.
Evaluation of White Lupin Germplasm for the Southeast
White lupin (Lupinus albus L.) has potential as a winter crop for grain, forage, cover crop, and wildlife browse in the southeastern USA, but adapted germplasm must be discovered and developed. Noffsinger and van Santen (1941–1950) examined 700 lines over 2 yr at two locations in central Alabama for agronomic traits and adaptation. Canonical discriminate analysis (CDA) was used to determine which agronomic traits and groups of accessions were most important for adaptation. Accessions from the East and West Mediterranean regions (vs. short season spring types from Southwest Asia and North Africa) were the best adapted.
Bindweed in a Wheat– Sunflower No-Tillage System
The objectives of a study by Jurado-Expósito et al. (2106–2112) were to assess and map population growth rate (PGR) of Convolvulus arvensis L. in a wheat (Triticum aestivum L.) and sunflower (Helianthus annuus L.) no tillage rotation over 4 yr and to determine the temporal stability of the distribution function of C. arvensis. Spatial variability of PGR was analyzed by geostatistics and temporal stability using a generalization of the two-sample Cramér–von Mises test for a difference between two univariate probability distributions. Year and crop influenced PGR, showing a moderate degree of aggregation in patches in both rotations. Overall, PGR became stable over the years. Knowledge of PGR could improve C. arvensis management if it were complemented with spatially herbicide targeted applications.
Yield Formation and Dry Matter in Soybean
Development of dry matter predictors for optimal yield would help soybean farmers identify profitability and aid in determination of environmental factors limiting yield. In this study, Board and Modali (1790–1799) collected data from studies involving a variety of cultural treatments and subjected yield, yield components, and dry matter relationships to regression analyses. The study indicated that 200 and 600 g m–2 at R1 and R5, respectively, were valid predictors for optimal yield. Environmental effects on yield were regulated by node number, pod number, and seed number per area.
Seed Processing and Native Grass Germination
Drought is cited as the most frequent cause of establishment failure of native plants. Springer (2075–2080) conducted an experiment to determine the interactions of seed processing and simulated dry seedbed conditions on the germination and seedling growth of five chaffy-seeded grasses that included: big bluestem (Andropogon gerardii Vitman), sand bluestem (Andropogon hallii Hack.), little bluestem [Schizachyrium scoparium (Michx.) Nash], yellow bluestem [Bothriochloa ischaemum (L.) Keng. var. ischaemum (Hack.) Celarier and Harlan] and indiangrass [Sorghastrum nutans (L.) Nash]. In general, seed germination and seedling growth were reduced as water potentials increased. Seed processing also affected the rate of seed germination. The germination and seedling growth of yellow bluestem was least affected by low water potentials or seed processing. Given the amount of genetic variation within most cross-pollinated species, this procedure could be used as a selection tool for breeding cultivars with improved germination and seedling growth at low water potentials.
Sustainable Grazing Systems in the Southern Great Plains
A primary goal of sustainable grazing systems in the southern Great Plains is to provide year-round high quality forage. Rao et al. (1973–1977) evaluated production patterns and nutritive values of the cool-season annual legumes grasspea (Lathyrus sativus L.) and lentil (Lens culinaris Med.) during May through July in central Oklahoma. Both cultivars produced high quality forage during three drought-affected growing seasons. Grasspea out-produced lentil and reached maximum yields (6415 vs. 2013 kg ha–1) by 75 d after seeding, while N concentration and IVDDM were similar. The higher level of production gives grasspea greater potential as a spring gap-filling forage in this region.
Wheat Cold Tolerance and Phospholipid Acyl Chains
Numerous changes occur in cell membrane components of winter wheat (Triticum aestivum L.) plants as they acclimate to cold temperatures. Skinner et al. (1858–1867) identified a previously unknown reengineering of the phospholipids, chemical compounds that comprise the majority of the cell membranes, as part of the cold acclimation process. The reengineering process results in a configuration of the membrane that imparts specific properties, probably contributing to cold tolerance. Gene expression changes that suggested involvement of phospholipids in signaling processes as part of the cold acclimation process were also observed.
Asymbiotic Reference Line Development by De Novo Mutagenesis
Determining the contribution of symbiotic nitrogen fixation to the yield formation in legume crops is a persisting demand that is still difficult to accomplish under field conditions. To solve this request for a particular pea (Pisum sativum cv. Bohat
r), Novák et al. (1837–1843) used de novo chemical mutagenesis to generate symbiotic mutants. They characterized mutant line Budfix2 (Nod+ Fix–) and showed it to be free of the pleiotropic effects of the symbiotic mutation on the plant growth and nitrogen metabolism. Budfix2 has a potential to be introgressed into other pea cultivars by backcrossing to create near-isogenic reference lines for the cultivars of interest.
Backcrossing in Cotton
Employing several novel approaches, Bayles et al. (2087–2095) compared six backcross family groups of cotton (Gossypium hirsutum L.) over multiple environments to measure the degree and rate of recovery of 14 recurrent parent (RP) traits. Reactions to three diseases were also determined for the Bc4F4 and the RP. Depending on the trait, three, four, and often more backcrosses were required to recover the RP traits. Several instances of transgressive segregation were noted mainly in the earlier backcross generations. The observed rate of recovery of RP traits was 96% of the theoretical rate. The backcrossed disease resistance tended to reflect that of its nonrecurrent parent. Resistance to the other two diseases was similar to the RP, but three possible exceptions were noted.
Soybean Breeding for Reduction of Linolenic Acid
Soybean seeds contain undesirable levels of oxidatively unstable linolenic acid in the oil fraction. Reduction of linolenic acid in oil by chemical hydrogenation results in the production of trans fatty acids. Low linolenic acid soybeans have been identified in mutagenesis breeding programs, but breeding for desired levels of linolenic acid is hampered by the complex nature of the trait. Bilyeu et al. (1830–1836) identified mutations in two desaturase genes underlying the trait in a low linolenic acid breeding line. Molecular markers specific for the mutant alleles were developed for more efficient incorporation of low linolenic acid into elite soybean varieties.
Modeling Spatial Correlation in Multienvironment Trials
Casanoves et al. (1927–1933) compared approaches to model within-trial spatial correlation, with and without heterogeneity of residual variances, for multi-environment trials (MET). Eighteen independent peanut (Arachis Hypogaea L.) METs were fitted by the classic ANOVA for a random block design and several mixed models including within-trial spatial correlation and between-trial heterogeneity. Results suggest that incorporating spatial dependence can improve mean comparisons in peanut METs, but it is not always better than models including random block effects. Even for the same crop, the choice of the best model may depend on other features, such as length of the growing season.
Genomic Constitution of Indian Sugarcane Cultivars
Sugarcane (Saccharum spp.) grows well under tropical conditions of the world. In India, systematic breeding initiated early in the twentieth century led to the development of cultivars suitable for subtropical conditions. In spite of a long breeding history, no systematic effort has been made to understand the genetic constitution of these cultivars. Selvi et al. (1750–1757) found that AFLP analysis revealed predominance (78.8%) of S. officinarum alleles those were equally shared by the 28 tropical and subtropical Indian cultivars. The subtropical cultivars, however, retained significantly higher number of S. spontaneum alleles than the tropical cultivars, reflecting the breeding strategy followed in the development of these cultivars. The level of genetic diversity between the tropical and subtropical cultivars was much higher than most of the pair wise diversity measures within each of these two adaptive groups. The AFLP-based clustering of the cultivars also corresponded well with their pedigree relationships.
Deploying rhg1 to Manage SCN
Soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) is the most damaging soybean [Glycine max (L.) Merr.] pest in the USA, and rhg1 is the most important SCN resistance gene in cultivars. In Brucker et al. (1721–1727), the rhg1 resistance allele originating from PI 88788 was tested in two near isogenic line (NIL) populations to determine its effect on SCN reproduction and agronomic traits. The rhg1 resistance allele had a dramatic effect on reducing the reproduction of HG type 0 SCN populations in the field and greenhouse, and in one NIL population, homozygous resistant lines out yielded the homozygous susceptible lines. These results demonstrate the benefits of deploying rhg1 to manage SCN.
Phosphorus Response of Brassica oleracea Genotypes
Crop cultivars selected to grow well on low P soils could improve the efficiency of P use. The objective of this study was to improve the selection criteria. Greenwood et al. (1728–1735) fitted a modified Michaelis–Menten equation to the relationships between shoot dry weight and P supply for 12 different Brassica oleracea L. genotypes grown in six field and glasshouse experiments. This equation fitted the data well. Ranking and regression analysis of the fitted parameters showed that intergenotypic variation in P response could be attributed largely to differences in maximal yield, a finding that could facilitate selection of B. oleracea genotypes for low P soils.
Intermountain West Forage Grasses Persistence
Excess soil salinity is a major factor limiting the establishment, persistence, and productivity of forage grasses in the Intermountain West. Within the cultivar NewHy, Jensen et al. (1717–1720) evaluated Cycles 1, 2, and 3 for increased plant persistence under increased salinity levels. After three cycles of selection, Cycle 3 was significantly more persistent than the original cultivar, NewHy. The largest gains were made during the first cycle of selection. Selection for persistence under saline conditions did not reduce seedling germination rate or vigor.
Pea Root Production and Architecture
Roots serve a significant role in plant growth; however, very little is known about root production and architecture because of difficulty associated with their study. McPhee (1758–1763) evaluated seedling roots of 330 pea (Pisum sativum L.) accessions for variation in overall root production and architectural differences. Significant variation was observed and a standard nomenclature was developed for pea to allow comparison among similar studies. Greater understanding of root architecture and the genetic basis of variation coupled with simple techniques for evaluation offer breeders and geneticists the opportunity to make improvements through selection.
Soybean Cumulative Light Interception
Yield responses for many crops under well-watered conditions have been considered as linearly related to the cumulative amount of light intercepted during the cropping season. Edwards et al. (1778–1785) evaluated a wide range of maturities and population densities in soybean [Glycine max (L.) Merr.] and found that yield increased nonlinearly with increasing cumulative light interception to reach an asymptote. Cumulative light interception to achieve near asymptotic yields could be manipulated by changing plant populations over a wide range of soybean maturities.
Soybean Yield and Biomass Responses to Increasing Plant Population
Edwards and Purcell (1770–1777) evaluated the soybean yield and biomass responses to increasing plant population among maturity group 00 through VI cultivars. Yield and biomass of all soybean maturity groups displayed an asymptotic response to increased plant population, and several of the maturity groups had similar asymptotic yield. The seeding density required to obtain 95% of asymptotic yield, however, varied among maturity groups and years, illustrating the difficulties encountered when attempting to make seeding rate recommendations using empirical relationships.
Agroforestry Herbage Utilization of Fertilizer Nitrogen
Herbage utilization of fertilizer N has not been adequately studied in agroforestry systems. Burner and MacKown (1817–1825) estimated N fertilizer recovery by herbage growing in an unshaded meadow and a shaded alley in a stand of 10-yr-old loblolly pine (Pinus taeda L.). Meadow herbage recovered nearly three times more fertilizer N than pine alley herbage. Low solar irradiance and depletion of soil water in July to September contributed to low fertilizer N recovery of herbage in the pine alley. Yearly applications of fertilizer N to similar sites probably should be limited to 
300 kg ha–1 yr–1 (meadow) and 100 kg ha–1 yr–1 (pine alley) for economic efficiency and to avoid undesirable consequences of excess mineral N accumulation in the soil.
Cultivated Timothy Has Potential Value for Rotational Grazing Systems
Cultivated timothy (Phleum pratense L.) germplasm has potential value for rotational grazing systems in North America. Casler (1990–1996) developed an improved population of timothy that had higher forage yield than commercial cultivars under both a frequent and an infrequent harvest management. Germplasm collected from old turf sods showed some potential for improving forage yield under frequent harvest management but was generally inferior to forage germplasm under infrequent harvest management. Timothy cultivars optimally adapted for use under rotational grazing will likely require different traits than traditional hay-type cultivars.
Classification of Maize Production Environments
Classification of maize (Zea mays L.) production environments enables investigation of the repeatable causes of genotype x environment interactions (GEI) for yield and agronomic traits to improve the predictability of cultivar performance. Löffler et al. (1708–1716) conducted crop simulation using the Crop Environment Resource Synthesis- Maize (CERES-Maize) model for each U.S. Corn Belt Township for 1952 through 2002 and for each of 266 trial locations in 2000 through 2002. Grain yield and biotic stress data for model confirmation were collected from 18 hybrids grown in replicated trials in those locations. Six major environment classes (EC) were identified with relative frequencies that varied greatly from year to year and which explained a significant component of the hybrid x EC interaction variance.
Optimal Soybean Seeding Population and Row Spacing
Soybean agronomists have long debated what factors govern the optimal seeding population and row spacing. In a seven-environment study, all with low yield, Morris and Kays (1826–1829) grew soybean [Glycine max (L.) Merr.] genotypes (MG III–MG VIII) varying in morphology to characterize cultivar x planting configuration interactions. One cultivar, AG3702, responded to increases in seeding population from 25 to 75 plants m–2 with a 47% seed yield increase, but nine others were relatively unresponsive. No cultivar x row spacing interactions were apparent, but seed yield in wide rows (71–76 cm) was equal to or greater than seed yield in narrow rows (18–35 cm). Our results suggested that the timing and severity of drought played a much larger role in a soybean cultivar's response to planting configuration than its morphology.
Defining Durum Wheat Adaptation Subregions
Annicchiarico et al. (1741–1749) assessed procedures for defining subregions and comparing wide- vs. specific-adaptation strategies for durum wheat (Triticum durum Desf.) in Algeria. An accurate and reliable subregion definition was obtained by interfacing site classification, based on genotype x location (GL) interaction, with a geographic information system (GIS). On the basis of observed and predicted yield gains, specific adaptation showed 2 to 7% greater gains vs. wide adaptation at similar costs. Specific adaptation in a subregion subject to stress may contribute to enhanced food security. Positive GL effects may also be exploited by national breeding programs in medium-sized countries.
Rhizome Expression in Birdsfoot Trefoil
Individual plant persistence of broadleaved birdsfoot trefoil (L. corniculatus L.) might be enhanced by rhizomes. Beuselinck et al. (1736–1740) evaluated rhizome expression of birdsfoot trefoil genotypes in latitudes of Iowa, Missouri, New York, Wisconsin, and Utah. Rhizomes appeared to be beneficial to plant survival and plant growth, but rhizomes did not assure performance or survival. The rhizomatous genotypes were larger and had greater mass than a nonrhizomatous cultivar, but high plant mortality and extensive crown and root necrosis reduced overall plant performance and rhizome expression. No single genotype performed well across locations. Interestingly, the Utah location was identified as having advantageous, but undefined, conditions beneficial to the performance of rhizomatous birdsfoot trefoil.
Russian Wheat Aphid Resistance
Since its introduction to the USA in 1987, the Russian wheat aphid [Diuraphis noxia (Mordvilko), RWA] has caused more than $1 billion in economic losses from yield reductions and pesticide inputs. In wheat (Triticum aestivum L.), host plant resistance to RWA had been highly effective in mitigating such losses until a new, virulent biotype (denoted as Biotype 2) was identified in Colorado in 2003. To identify germplasm with resistance to Biotype 2 RWA, Collins et al. (1877–1880) evaluated a collection of 761 wheat germplasm accessions that had been previously shown to carry resistance to Biotype 1 RWA. Ten accessions with a very high level of resistance were identified. These accessions will be extremely useful for genetics studies and cultivar development.
Sunflower Nuclear Male Sterility Gene Mapping
The nuclear male sterility genes Ms10 and Ms11 have been mapped in sunflower (Helianthus annuus L.) by using genetic maps constructed with simple sequence repeat or microsatellite (SSR), insertion–deletion polymorphism, restriction fragment length polymorphism, or single nucleotide polymorphism markers. Ms10 maps to Linkage Group (LG) 11, and is tightly linked to four SSR markers (ORS697, ORS1214, ORS686, CRT162). Ms11 maps to LG8, and is flanked by the SSR markers MS925 and ORS536. The availability of tightly linked polymerase chain reaction based markers and the location of the Ms10 and Ms11 genes on the sunflower genetic map will be useful for marker-assisted selection and provides a basis for the physical mapping and positional cloning of these genes. See Pérez-Vich et al. (1851–1857).
Nondestructive Measurement of Carotenoids in Plant Tissues
Genetic selection for carotenoids in rice bran would be facilitated by the development of a nondestructive technique to screen individual kernels. Belefant-Miller et al. (1786–1789) developed a fluorescence quenching technique to measure carotenoid levels that takes advantage of two factors: the tendency of plant tissues to fluoresce when excited by specific light wavelengths and the tendency of carotenoids to quench this fluorescence. Utilizing this technique, the presence of carotenoids in rice bran was found to vary by cultivar. Since the technique is nondestructive and requires only a very small sample, measurements were also able to be made on living plants and within the different colors of a variegated leaf.
Temporal Distribution of Soybean Pod Set
The temporal distribution of pod production and survival play an important role in determining pod number in soybean [Glycine max (L.) Merrill]. Egli and Bruening (1764–1769) investigated the effect of reducing photosynthesis with shade on these distributions. Continuous shade reduced pod number, but shortened the pod production period in only one of four comparisons. Pod production increased quickly when shade was removed midway through flowering, but pod number at maturity was still less than the control. These results suggest that maximum pod number in soybean may require a stress-free environment throughout the entire flowering and pod set period.
Genome Size Evolution in the Grasses
The grasses show striking differences in the size of their genomes. The evolutionary implication of these differences is important given that conservation of gene content and order has been used to extend genomic information from one species (e.g., rice, Oryza sativa L.) to others. Understanding how genome size changes during evolution requires knowledge of phylogenetic relationships within the grasses. In a study by Caetano-Anollés (1809–1816), a phylogeny of the grasses integrated from DNA sequences, RNA structure, large-scale chromosomal rearrangements, and morphological data was used to trace the evolution of genome size using well-established methods of character state reconstruction. While an overall pattern of genome size increase was evident, it appears that genome size has both increased and decreased along every grass lineage in the course of evolution. According to this study, the genome of the ancestor of the grass family had 3.0 to 5.2 pg DNA per 2C nucleus.
QTL Analysis of White Clover Seed Production
Use of marker-assisted selection (MAS) in open-pollinated species may accelerate improvement of forages with respect to economic and environmental performance measures. White clover (Trifolium repens L.) seed yield is highly heritable, yet extremely variable in populations selected on forage performance characteristics, creating an excellent case-study in MAS development for out-crossing species. The first step in MAS is the understanding of genetic architecture of the target trait and the establishment of robust associations between markers and plant performance. Reporting from New Zealand, Barrett et al. (1844–1850) detail the discovery of QTLs associated with seed production in white clover.
Potential Heterotic Groups in Hop
Hop (Humulus lupulus L.) breeders have historically used hybridization and clonal selection to develop new cultivars. The use of genetically diverse parents to capture heterosis in the offspring has received little attention in hop breeding. Townsend and Henning (1901–1907) used AFLP molecular markers to characterize the genetic relationship among 106 diverse hop genotypes. Thirteen potential heterotic groups were identified among the genotypes studied. These results suggest potential parental combinations for capturing heterosis in hop.
Related articles in Crop Science:
r by De Novo Mutagenesis
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