A number of attributes make alfalfa an attractive crop for production of biofuels and for biorefining. Alfalfa has a long history of cultivation around the world. It was introduced several times into North America during the 1700s and 1800s and is currently grown across the continent (Russelle, 2001). In the United States, alfalfa is the fourth most widely grown crop with over 9.3 million hectares of alfalfa harvested in 2003 (USDA-NASS, 2004). It is a perennial plant that is typically harvested for four years (an establishment year plus three subsequent years). Depending on location, alfalfa is harvested three or more times each year by cutting the stems near ground level. On average across the United States, alfalfa yields 7.8 Mg of dry matter (DM) per hectare each year, although yields can vary by location from 3.4 (North Dakota) to 18.4 (Arizona) Mg ha-1 (USDA- NASS, 2004). In 2003 the national harvest of alfalfa was over 69 million metric tons (USDA-NASS, 2004). The technology for cultivation, harvesting, and storing alfalfa is well established, machinery for harvesting alfalfa is widely available, and farmers are familiar with alfalfa production. There is a well-developed indus­try for alfalfa cultivar development, seed production, processing, and distribution. Alfalfa breeders have utilized the extensive germplasm resources of alfalfa to introduce disease and insect resistance, expand environmental adaptation, and improve forage quality. Nonetheless, alfalfa cultivation requires fertile, deep, well-drained soils of near neutral pH and is limited to humid areas with adequate rainfall. In arid or semi-arid areas, irrigation is essential for crop production. Despite breeding efforts that have increased disease and pest resistance, alfalfa yields have not increased substantially over the past 25 years (Brummer, 1999).

The high biomass potential of alfalfa is based on underground, typically unobserved traits. Alfalfa develops an extensive, well-branched root system that is capable of penetrating deep into the soil. Root growth rates of 1.8 m a year are typical in loose soils (Johnson et al., 1996) and metabolically active alfalfa roots have been found 18 m or more below ground level (Kiesselbach et al., 1929). This deep root system allows alfalfa plants to access water and nutrients that are not available to more shallowly rooted annual plants, which enables established alfalfa plants to produce adequate yields under less than optimal rainfall conditions. Alfalfa roots engage in a symbiotic relationship with the soil bacterium Sinorhizobium meliloti. This partnership between the plant and bacterium results in the formation of a unique organ, the root nodule, in which the bacterium is localized. The bacteria in root nodules take up nitrogen gas (N2) and “fix” it into ammonia. The ammonia is assimilated through the action of plant enzymes to form glutamine and glutamate. The nitrogen-containing amide group is subsequently transferred to aspartate and asparagine for transport throughout the plant. On average, alfalfa fixes approximately 152 kg N2 ha-1 on an annual basis as a result of biological nitrogen fixation, which eliminates the need for applied nitrogen fertilizers (Russelle and Birr, 2004). Although a signif­icant proportion of the fixed nitrogen is removed by forage harvest, fixed nitrogen is also returned to the soil for use by subsequent crops. This attribute of increasing soil fertility has made alfalfa and other plants in the legume family crucial components of agricultural systems worldwide. Cultivation of alfalfa has also been shown to improve soil quality, increase organic matter, and promote water penetration into soil.

Responsible stewardship of agricultural lands has never been more important. Utilization of alfalfa as a biomass crop has numerous environmental advantages. There is an urgent need to increase the use of perennials in agricultural systems to decrease erosion and water contamination. Annual row crop production has been shown to be a major source of sediment, nutrient (nitrogen and phosphorus), and pesticide contamination of surface and ground water. Perennial crops such as alfalfa can reduce the nitrate concentrations in soil and drainage water, and prevent soil erosion (Huggins et al., 2001). In addition, energy costs associated with production of alfalfa are low. A recent study shows that energy inputs for production of alfalfa are far lower than for production of corn and soybean, and very similar to switchgrass (Kim and Dale, 2004), primarily because alfalfa does not require nitrogen fertilizer. Biorefining could increase the return on alfalfa production so that cultivation of the crop is more economically attractive, as well as environmentally beneficial.

An additional advantage of using alfalfa for biofuel production compared to other crops is the ability to easily separate leaves and stems to produce co­products. In fact, alfalfa herbage can almost be considered two separate crops because leaves and stems differ so dramatically in composition. On a dry weight basis, total alfalfa herbage contains 18-22% protein with leaves containing 26-30% protein and stems only 10-12% (Arinze et al., 2003). In some analyses, alfalfa protein has been valued highly, theoretically greatly reducing the cost of the lignocelluose fraction (Dale, 1983). Several different integrated processes for refining alfalfa have been proposed based primarily on the method of refining the protein fraction. From field-dried hay, leaves may be separated from stem material mechanically (see “Protein and Fiber Separation” below). The leaf meal could be used as a high-protein feed with the stems utilized for gasification and conversion to electricity (Downing et al., 2005) or fermentation to ethanol (Dale, 1983). Alternatively, protein could be extracted from total ground material and the residue used for fermentation. Fresh forage can be “juiced” to remove protein and the residue fermented to ethanol or other products (Koegel et al., 1999; Sreenath et al., 2001; Weimer et al., 2005). An economic analysis of these

alternatives is beyond the scope of this chapter. However, a comparison of the potential costs and revenues of different biobased feedstocks to produce ethanol and other products is clearly needed to advance biomass refining from the theo­retical to practical stages.

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