Evaluation of Sweet Sorghum and Sorghum × Sudangrass Hybrids as Feedstocks for Ethanol Production

Field studies were conducted at the USDA-ARS Sugarcane Research Laboratory in southeast Louisiana to evaluate the ethanol yield potential of five sweet sorghums (Dale, M81-E, Rio, Theis, and Topper) and two non-flowering sorghum × sudangrass forage hybrids (MMR 333/27 and MMR 333/47). The sorghums were planted in the spring and harvested at 85, 101, 119, and 138 days after planting (DAP). Theoretical sugar-based ethanol yield increased for the sweet sorghums (except Rio) from 85 through 119 days, but did not significantly increase further at 138 days. The forage sorghums did not show a similar increase, though the theoretical sugar-based ethanol yield of MMR 333/47 at 138 DAP was greater than at 85 DAP. Conversely, theoretical fiber-based ethanol yields increased two-fold in the two forage sorghums from 85 to 138 DAP; a significant increase in fiber-based ethanol yield was not observed in any of the sweet sorghums over the same period. At 138 DAP, sugar-based ethanol yield of Theis (6,060 L ha^?1), was greater than that of Rio or either of the two forage hybrids. Fiber-based ethanol yield of MMR 333/47 (8,860 L ha^?1) was greater than that of any other variety in the test. Theoretical ethanol yield from hexose sugar and fiber components averaged across varieties was 6,500, 7,720, 9,100, and 10,810 L ha^?1 at 85, 101, 119 and 138 DAP, respectively. As a complementary crop for Louisiana’s sugarcane growers, sorghum would need to be harvested not later than 120 DAP so as to not interfere with the planting of sugarcane in these fields. Both Theis and MMR 333/47 produced greater than 11,000 L ha^?1 combined theoretical ethanol at 119 DAP, Theis, equally from sugar and fiber, MMR 333/47 about two-thirds from fiber. Choice of sorghum type would depend on the conversion process(s) being used at the biorefinery.     

Assessment of Sugarcane Billet Harvester on Recovery of Sweet Sorghum Biomass for Ethanol Production

Sweet sorghum (Sorghum bicolor L. Moench) is a promising bioenergy crop for the production of ethanol and bio-based products. Sugarcane billet harvesters can be used to harvest sweet sorghum. Multiple extractor fan speed settings of these harvesters allow for separating the extraneous matter in the feedstock, which has been associated with increased milling throughput and better juice quality at the processing facility. This removal is not completely selective, and some stalk material is also lost. These losses can be higher for sweet sorghum than sugarcane due its lower weight. This paper presents an assessment of how the speed of the primary extractor fan of a sugarcane billet combine used for harvesting sweet sorghum affects the biomass yield, biomass losses, and quality at delivery for the production of ethanol from extracted juice and fiber. Three primary extractor fan speeds (0, 800, and 1100 rpm) were evaluated. Higher fan speeds decreased fresh biomass yields by up to 28.3 Mg ha^?1. Juice quality was not significantly different among treatments. Ethanol yield calculated from sweet sorghum harvested at 0 rpm was 6075 L ha^?1. This value decreased by about half for material harvested at 1100 rpm due to the differences in biomass yield.     

Evaluation of Public Sweet Sorghum A-Lines for Use in Hybrid Production

A fundamental need for commercialization of sweet sorghum [Sorghum bicolor (L.) Moench] as a bioenergy crop is an adequate seed supply, which will require development of hybrid varieties using dwarf seed-parent lines. A set of six public sweet sorghum A-lines (Dwarf Kansas Sourless, KS9, N36, N38, N39, and N4692) were crossed with a set of six public sweet sorghum cultivars (Brawley, Kansas Collier, Dale, Sugar Drip, Waconia, and Wray). Grain, fiber, and sugar yields were determined, and conversion formulas were applied to estimate ethanol yields. Hybrids were grown in fields at Ithaca, NE, USA, in 1983–1984 fertilized with 112 kg ha^?1 N. In terms of yield components and overall ethanol yields, one A-line, N38, was inferior. Average total ethanol yields from hybrids made on the other A-lines were not significantly different, suggesting that any of those five A-lines could be useful seed-parents. With the exception of grain yield, cultivars used as pollen parents were among the highest-performing entries for all traits. For all traits directly contributing to total ethanol yield (grain yield, juice yield, % soluble solids, sugar yield, fiber yield), hybrids were also among the highest-performing entries. Results of this study demonstrate that hybrid sweet sorghum with performance criteria equivalent to existing sweet sorghum cultivars can be produced on the sweet sorghum seed-parent lines A-Dwarf Kansas Sourless, A-KS9, A-N36, A-N39, and A-N4692. Identification of specific seed-parent × pollen parent lines with characteristics best suited for particular growing regions and end-user needs will be critical for commercial hybrid development.     

Genetic transformation of sweet sorghum

Sweet sorghum has substantial potential as a biofuel feedstock, with advantages in some environments over alternatives such as sugarcane or maize. Gene technologies are likely to be important to achieve yields sufficient for food, fuel and fibre production from available global croplands, but sorghum has proven difficult to transform. Tissue culture recalcitrance and poor reproducibility of transformation protocols remain major challenges for grain sorghum, and there has been no reported success for sweet sorghum. Here we describe a repeatable transformation system for sweet sorghum, based on (1) optimized tissue culture conditions for embryogenic callus production with >90% regenerability in 12-week-old calli, and (2) an effective selection regimen for hygromycin resistance conferred by a Ubi-hpt transgene following particle bombardment. Using this method, we have produced sixteen independent transgenic lines from multiple batches at an overall efficiency of 0.09% transformants per excised immature embryo. Co-expression frequency of a non-selected luciferase reporter was 62.5%. Transgene integration and expression were confirmed in T₀ and T₁ plants by Southern analysis and luciferase assays. This success using the major international sweet sorghum cultivar Ramada provides a foundation for molecular improvement of sweet sorghum through the use of transgenes. Factors likely to be important for success with other sweet sorghum cultivars are identified.     

Quantitative trait loci for cell-wall components in recombinant inbred lines of maize (Zea mays L.) I: stalk tissue

Maize silage is a significant energy source for animal production operations, and the efficiency of the conversion of forage into animal mass is an important consideration when selecting cultivars for use as feed. Fiber and lignin are negatively correlated with digestibility of feed, so the development of forage with reduced levels of these cell-wall components (CWCs) is desirable. While variability for fiber and lignin is present in maize germplasm, traditional selection has focused on the yield of the ear rather than the forage quality of the whole plant, and little information is available concerning the genetics of fiber and lignin. The objectives of this study were to map quantitative trait loci (QTLs) for fiber and lignin in the maize stalk and compare them with QTLs from other populations. Stalk samples were harvested from 191 recombinant inbred lines (RILs) of B73 (an inbred line with low-to-intermediate levels of CWCs) x De811 (an inbred line with high levels of CWCs) at two locations in 1998 and one in 1999 and assayed for neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent lignin (ADL). The QTLs were detected on nine chromosomes, mostly clustered in concordance with the high genetic correlations between NDF and ADF. Adjustment of NDF for ADF and ADF for ADL revealed that most of the variability for CWCs in this population is in ADF. Many of the QTLs detected in this study have also been detected in other populations, and several are linked to candidate genes for cellulose or starch biosynthesis. The genetic information obtained in this study should be useful to breeding efforts aimed at improving the quality of maize silage.     

Morphological and molecular characterization of sweet,grain and forage sorghum (Sorghum bicolor L.) genotypes grown under temperate climatic conditions

Abstract

In the present study, we used 12 genotypes of sorghum originated from different countries (five sweet, four grain and three forage). These different genotypes and types of sorghum were evaluated for the agro-morphological traits that are associated with the estimated sugar and bioethanol yield to estimate their phenotypic diversity. Analysis of variance showed significant differences between different types of sorghum for all the evaluated traits. Sweet sorghum genotypes, however, showed better performance with respect to all studied traits than the other genotypes. A positive significant correlation was observed between plant height, leaf number, leaf area, biomass yield, cane and bagasse yields, and the predicted bioethanol yield. Both, cluster and principal component analysis were performed to group the genotypes according to their agro-morphological and molecular similarity coefficients. For analytical approaches, the Iranian grain and forage genotypes clustered separately from the other genotypes. The clustering patterns obtained from the molecular dominant markers had higher discriminatory power than using morphological characters to separate sweet genotypes from the forage and grain sorghum ones. The results clearly indicated that sweet sorghum can be grown in Germany and maintains its superiority in biomass production and sugar yield over grain and forage sorghum types.     

Bagasse Silage from Sweet Pearl Millet and Sweet Sorghum as Influenced by Harvest Dates and Delays between Biomass Chopping and Pressing

Bagasse remaining after extracting the juice from crop biomass for ethanol production could be preserved as silage and used in animal feedstock, but the nutritive and conservation attributes of bagasse silage from sweet sorghum (Sorghum bicolor (L.) Moench) and sweet pearl millet (Pennisetum glaucum (L.) R.Br) are not well known. We evaluated the nutritive and conservation attributes of silages made with the bagasse of two species (sweet pearl millet and sweet sorghum) harvested on two dates (August and September) at two sites in Québec (Canada) and ensiled after four delays between biomass chopping and pressing (0.5, 2, 4, and 6 h). Bagasse silages made in laboratory silos were considered well preserved (pH?≤?4.0, NH₃-N?<?100 g kg^?1 total N, lactate?>?30 g kg^?1 DM, no propionic and butyric acids) regardless of species, harvest date, or delay between biomass chopping and pressing. Sweet pearl millet and sweet sorghum bagasse silages had similar total N concentration, in vitro true digestibility of dry matter (IVTD), and in vitro neutral detergent fiber digestibility (NDFD). Bagasse silage made from biomass harvested in August rather than in September had a 4 % greater concentration of total N, a 4 % greater IVTD, and a 8 % greater NDFD. The delay between biomass chopping and pressing did not affect the nutritive and conservation attributes of silages. Juice extraction from the biomass of sweet pearl millet and sweet sorghum did not impair attributes of good silage fermentation but it reduced its nutritive value.     

Biorefinery of sweet sorghum stem

Sweet sorghum has been considered as a viable energy crop for alcohol fuel production. This review discloses a novel approach for the biorefining of sweet sorghum stem to produce multiple valuable products, such as ethanol, butanol and wood plastic composites. Sweet sorghum stem has a high concentration of soluble sugars in its juice, which can be fermented to produce ethanol by Saccharomyces cerevisiae. In order to obtain high ethanol yield and fermentation rates, concentrated juice with an initial total sugar concentration of 300gL(-1) was fermented. The maximum ethanol concentration after 54h reached 140gL(-1) with a yield of 0.49g ethanol per g consumed sugar, which is 97% of the theoretical value. Sweet sorghum bagasse, obtained from juice squeezing, was pretreated by acetic acid to hydrolyze 80-90% of the contained hemicelluloses. Using this hydrolysate as raw material (total sugar 55gL(-1)), 19.21gL(-1) total solvent (butanol 9.34g, ethanol 2.5g, and acetone 7.36g) was produced by Clostridium acetobutylicum. The residual bagasse after pretreatment was extruded with PLA in a twin-screw extruder to produce a final product having a PLA: fiber ratio of 2:1, a tensile strength of 49.5M and a flexible strength of 65MPa. This product has potential use for applications where truly biodegradable materials are required. This strategy for sustainability is crucial for the industrialization of biofuels from sweet sorghum.     

Agronomic Considerations for Sweet Sorghum Biofuel Production in the South-Central USA

Sweet sorghum (Sorghum bicolor (L.) Moench) is currently recognized throughout the world as a highly promising biomass energy crop. Production systems and management practices for sweet sorghum have not been fully developed for the USA, although sporadic research efforts during recent decades have provided some insights into production of sweet sorghum primarily for fermentable sugar production. Field plot experiments were conducted at sites across Louisiana to assess biomass and sugar yield responses to N fertilizer, plant density, and selected cultivars. Although linear increases in stem biomass production and fermentable sugar yield were obtained with increasing N fertilizer rate under irrigated conditions, most of the increase was from the initial 45 kg N ha⁻¹ increment. Nitrogen fertilization increased stem biomass production but not fermentable sugar yield in some non-irrigated environments. Increased plant density contributed to fermentable sugar yield only under growth-limiting conditions, particularly under limited soil moisture. Location effects indicate that sweet sorghum may not be suitable for some sub-optimal cropland and pasture environments in Louisiana. During the primary growing season, cultivar did not affect fermentable sugar yields, although Dale was consistently high in sugar concentration during this period. Nitrogen fertilizer increased fermentable sugar yields only when moisture was not limiting. Overall results indicate that in environments where soil moisture limits plant growth, sugar yield responses are likely from increased plant density and not from increased N fertilization.     

Production, transportation and milling costs of sweet sorghum as a feedstock for centralized bioethanol production in the upper Midwest

Sweet sorghum has been identified as a possible ethanol feedstock because of its biomass yield and high concentration of readily fermentable sugars. It has found limited use, however, because of poor post-harvest storage characteristics and short harvest window in cooler climates. Previous research (Bennett, A.S., Anex, R.P., 2008. Farm-gate production costs of sweet sorghum as a bioethanol feedstock. Transactions of the ASABE 51(2), 603-613) indicates that fermentable carbohydrates (FC) can be produced at less expense from sweet sorghum than from corn grain. Previous research, however, did not include costs associated with off-farm transportation, storage, or capital costs associated with milling and energy recovery equipment that are required to provide FC suitable for biological conversion. This study includes these additional costs and reevaluates sweet sorghum as a biocommodity feedstock. A total of eight harvest-transport-processing options are modeled, including 4-row self-propelled and 2-row tractor-pulled forage harvesters, two different modes of in-field transport, fresh processing, on-farm ensilage and at-plant ensilage. Monte Carlo simulation and sensitivity analysis are used to account for system variability and compare scenarios. Transportation costs are found to be significant ranging from $33 to $71 Mg (-1) FC, with highest costs associated with at-plant ensilage scenarios. Economies of scale benefit larger milling equipment and boiler systems reducing FC costs by more than 50% when increasing annual plant capacity from 37.9 to 379 million liters. Ensiled storage of high moisture sweet sorghum in bunkers can lead to significant losses of FC (>20%) and result in systems with net FC costs well above those of corn-derived FC. Despite relatively high transport costs, seasonal, fresh processed sweet sorghum is found to produce FC at costs competitive with corn grain derived FC.     

Composition of sugar cane,energy cane,and sweet sorghum suitable for ethanol production at Louisiana sugar mills

A challenge facing the biofuel industry is to develop an economically viable and sustainable biorefinery. The existing potential biorefineries in Louisiana, raw sugar mills, operate only 3 months of the year. For year-round operation, they must adopt other feedstocks, besides sugar cane, as supplemental feedstocks. Energy cane and sweet sorghum have different harvest times, but can be processed for bio-ethanol using the same equipment. Juice of energy cane contains 9.8% fermentable sugars and that of sweet sorghum, 11.8%. Chemical composition of sugar cane bagasse was determined to be 42% cellulose, 25% hemicellulose, and 20% lignin, and that of energy cane was 43% cellulose, 24% hemicellulose, and 22% lignin. Sweet sorghum was 45% cellulose, 27% hemicellulose, and 21% lignin. Theoretical ethanol yields would be 3,609 kg per ha from sugar cane, 12,938 kg per ha from energy cane, and 5,804 kg per ha from sweet sorghum.     

Sugarcane Internode Composition During Crop Development

Sugarcane sugar and bagasse can be utilized for the production of ethanol or other biofuels. A better understanding of the changes in composition with development along the stalk and with crop development will maximize the usage of sugarcane for this purpose. Two experiments were designed to elucidate internode composition changes during the growing season. In experiment 1, an internode of stalks of 5 modern cultivars were marked at the start of elongation, and then sampled every 1 to 2?weeks from July until October. Sugars were extracted and assayed, and a sequential detergent method was used to estimate hemicellulose, cellulose, and lignin contents. In experiment 2, internodes 1, 3, 5, 7, 9, and 11 down the stalk were sampled in late July (grand growth) and late September (ripening). Internode length, fresh weight, dry weight, water content, and sugar contents were determined as well as cell wall composition. Both experiments were repeated in 2?years. As internodes elongated, total sugar increased, and hemicellulose decreased as a proportion of neutral detergent fiber, while cellulose and lignin increased. After elongation, sucrose and lignin increased, and cellulose content decreased with internode age. The variability in cell wall composition among the five cultivars suggests that selection for desirable composition may be possible. In Experiment 2, hemicellulose contents were lower, and lignin and ash contents were higher at ripening than during grand growth. Delaying sugarcane harvest to maximize sucrose content may decrease bagasse suitability for cellulosic ethanol production because of the increased lignin content.     

Low temperature alkali pretreatment for improving enzymatic digestibility of sweet sorghum bagasse for ethanol production

A low temperature alkali pretreatment method was proposed for improving the enzymatic hydrolysis efficiency of lignocellulosic biomass for ethanol production. The effects of the pretreatment on the composition, structure and enzymatic digestibility of sweet sorghum bagasse were investigated. The mechanisms involved in the digestibility improvement were discussed with regard to the major factors contributing to the biomass recalcitrance. The pretreatment caused slight glucan loss but significantly reduced the lignin and xylan contents of the bagasse. Changes in cellulose crystal structure occurred under certain treatment conditions. The pretreated bagasse exhibited greatly improved enzymatic digestibility, with 24-h glucan saccharification yield reaching as high as 98% using commercially available cellulase and β-glucosidase. The digestibility improvement was largely attributed to the disruption of the lignin-carbohydrate matrix. The bagasse from a brown midrib (BMR) mutant was more susceptible to the pretreatment than a non-BMR variety tested, and consequently gave higher efficiency of enzymatic hydrolysis.     

Improvement of biomass through lignin modification

Lignin, a major component of the cell wall of vascular plants, has long been recognized for its negative impact on forage quality, paper manufacturing, and, more recently, cellulosic biofuel production. Over the last two decades, genetic and biochemical analyses of brown midrib mutants of maize, sorghum and related grasses have advanced our understanding of the relationship between lignification and forage digestibility. This work has also inspired genetic engineering efforts aimed at generating crops with altered lignin, with the expectation that these strategies would enhance forage digestibility and/or pulping efficiency. The knowledge gained from these bioengineering efforts has greatly improved our understanding of the optimal lignin characteristics required for various applications of lignocellulosic materials while also contributing to our understanding of the lignin biosynthetic pathway. The recent upswing of interest in cellulosic biofuel production has become the new focus of lignin engineering. Populus trichocarpa and Brachypodium distachyon are emerging as model systems for energy crops. Lignin research on these systems, as well as on a variety of proposed energy crop species, is expected to shed new light on lignin biosynthesis and its regulation in energy crops, and lead to rational genetic engineering approaches to modify lignin for improved biofuel production.     

Characterization of novel Brown midrib 6 mutations affecting lignin biosynthesis in sorghum

The presence of lignin reduces the quality of lignocellulosic biomass for forage materials and feedstock for biofuels. In C4 grasses,the brown midrib phenotype has been linked to mutations to genes in the monolignol biosynthesis pathway. For example,the Bmr6 gene in sorghum(Sorghum bicolor) has been previously shown to encode cinnamyl alcohol dehydrogenase(CAD),which catalyzes the final step of the monolignol biosynthesis pathway. Mutations in this gene have been shown to reduce the abundance of lignin,enhance digestibility,and improve saccharification efficiencies and ethanol yields. Nine sorghum lines harboring five different bmr6 alleles were identified in an EMS-mutagenized TILLING population. DNA sequencing of Bmr6 revealed that the majority of the mutations impacted evolutionarily conserved amino acids while three-dimensional structural modeling predicted that all of these alleles interfered with the enzyme's ability to bind with its NADPH cofactor. All of the new alleles reduced in vitro CAD activity levels and enhanced glucose yields following saccharification. Further,many of these lines were associated with higher reductions in acid detergent lignin compared to lines harboring the previously characterized bmr6-ref allele. These bmr6 lines represent new breeding tools for manipulating biomass composition to enhance forage and feedstock quality.     

A Novel Wild-Type Saccharomyces cerevisiae Strain TSH1 in Scaling-Up of Solid-State Fermentation of Ethanol from Sweet Sorghum Stalks

The rising demand for bioethanol, the most common alternative to petroleum-derived fuel used worldwide, has encouraged a feedstock shift to non-food crops to reduce the competition for resources between food and energy production. Sweet sorghum has become one of the most promising non-food energy crops because of its high output and strong adaptive ability. However, the means by which sweet sorghum stalks can be cost-effectively utilized for ethanol fermentation in large-scale industrial production and commercialization remains unclear. In this study, we identified a novel Saccharomyces cerevisiae strain, TSH1, from the soil in which sweet sorghum stalks were stored. This strain exhibited excellent ethanol fermentative capacity and ability to withstand stressful solid-state fermentation conditions. Furthermore, we gradually scaled up from a 500-mL flask to a 127-m³ rotary-drum fermenter and eventually constructed a 550-m³ rotary-drum fermentation system to establish an efficient industrial fermentation platform based on TSH1. The batch fermentations were completed in less than 20 hours, with up to 96 tons of crushed sweet sorghum stalks in the 550-m³ fermenter reaching 88% of relative theoretical ethanol yield (RTEY). These results collectively demonstrate that ethanol solid-state fermentation technology can be a highly efficient and low-cost solution for utilizing sweet sorghum, providing a feasible and economical means of developing non-food bioethanol.     

High-Throughput Profiling of the Fiber and Sugar Composition of Sugarcane Biomass

Lignocellulosic biomass from sugarcane (Saccharum spp. hybrids) could potentially be a major feedstock for second-generation biofuel production. Consequently, selecting sugarcane varieties with favorable biomass characteristics, typically less enzymatic recalcitrance and better saccharification yield without sugar-yield penalty, will be important in sugarcane breeding. Economical and high-throughput techniques for profiling the major biomass components of this complex system will facilitate selection of clones with ideal lignocellulosic composition from large numbers of genotypes in breeding programs. We used a combined high-throughput profiling approach to evaluate the biomass composition of samples from a sugarcane germplasm collection. This employed near-infrared (NIR) spectroscopy for fiber characterization and high-performance liquid chromatography (HPLC) for determining the sugar content in juice. The results for 331 samples, from a diverse sugarcane population of 186 genotypes, derived from 143 parents of different genetic backgrounds, showed that high-quality NIR spectroscopic predictions were feasible for cellulose, hemicellulose, lignin, and extractives values in fiber, and sugars in juice were suitably analyzed by HPLC. The analysis of total biomass indicated that this NIR- and HPLC-based high-throughput method allowed a robust phenotypic assessment of a large number of samples for the key biomass traits in the sugarcane system, including total dry biomass, fiber, sugar content, and theoretical ethanol yields, and could potentially become the method of choice for sugarcane germplasm screening in breeding programs targeting the support of biofuel production.     

Effects of Forage Quality and Cell Wall Constituents of Bermuda Grass on Biochemical Conversion to Ethanol

Bermuda grass is an attractive candidate as a feedstock for biofuel production because over four million hectares of Bermuda grass are already grown for forage in the Southern USA. Because both rumen digestion and biochemical conversion to ethanol depend upon enzymatic conversion of the cell wall polysaccharides into fermentable sugars, it is probable that grasses bred for increased forage quality would be more amenable for ethanol production. However, it is not known how variation in rumen digestibility and cell wall/fiber components correlates with efficiency of conversion to ethanol via fermentation. The objective of this research was to determine relationships between ethanol production evaluated by simultaneous saccharification and fermentation (SSF), 72-h in vitro ruminal dry matter digestibility (IVDMD), in vitro ruminal gas production after 24 and 96 h, and biomass composition for 50 genetically diverse Bermuda grass accessions. The Bermuda grass samples were subjected to standard 72-h IVDMD and forage fiber analyses. Also, in separate labs, gas production was measured in sealed volume-calibrated vials after 24 (NNG24) and 96 h (NNG96) of in vitro fermentation by ruminal fluid; ethanol and pentose sugar productions were measured from a bench-top SSF procedure; cell wall constituents were determined by the Uppsala Dietary Fiber Method; and total nitrogen, carbon, and ash concentrations were determined by using the LECO combustion method. Ethanol production was moderately correlated with IVDMD (r?=?0.55) and NNG96 (r?=?0.63) but highly correlated with NNG24 (r?=?0.93). Ethanol was negatively correlated with neutral detergent fiber (NDF; r?=??0.53) and pentose sugars (r?=??0.60), but not correlated with glucose content. Regression models indicated that NDF and cell wall pentose sugar concentrations had significant negative effects on ethanol production. Variation among entries for IVDMD was affected by variability of NDF, pentose sugar concentrations, and biomass nitrogen content. Variation in Klason lignin content had only minor negative impacts on ethanol production and IVDMD. Biochemical conversion efficiency of Bermuda grass by SSF can be best estimated by NNG24 but not by IVDMD.     

QTL analyses of fiber components and crude protein in an annual × perennial ryegrass interspecific hybrid population

Annual (Lolium multiflorum Lam.) and perennial (Lolium perenne L.) ryegrasses are two important forage and turfgrass species. Improving the digestibility of forage by decreasing fiber content is a major goal in forage crop breeding programs. An annual × perennial ryegrass interspecific hybrid population was used to map quantitative trait loci (QTLs) for fiber components, neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent lignin (ADL), and crude protein (CP). Samples were harvested three times in August and September 2003 and August 2004, respectively. Simple interval mapping was used to detect QTLs from both the male and female parental maps previously developed for the population. Fiber components were all correlated positively with each other and were negatively correlated with CP. The largest correlations were between NDF and ADF with r = 0.86, 0.72, and 0.82 for each of the three harvests. All four traits showed intermediate broad-sense heritability values ranging from 0.35 to 0.72. A total of 63 QTLs were detected for the four traits measured over the three harvests from both the female and male maps. Coincident QTLs were detected on linkage groups (LGs) 2, 6, and 7 for NDF, LGs 1, 2, and 7 for ADF, LGs 6 and 7 for ADL, and LG 2 for CP, respectively. Coincident QTLs were also detected on LGs 2, 6, and 7 for NDF and ADF, providing evidence of the genetic basis of the observed high level of phenotypic correlation. The QTLs on LGs 2, 6, and possibly 7 for fiber components were co-located on the same LG as several lignin biosynthetic genes from perennial ryegrass.