A system for identifying and quantifying the stages of growth and development of perennial forage grasses was developed. The system consists of a universal set of morphological descriptors for forage and range grasses and a continuous numerical index. The life cycle of individual grass tillers is divided into five primary growth stages (i) germination, (ii) vegetative, (iii) elongation, (iv) reproductive, and (v) seed ripening. Substages corresponding to specific morphological events are defined within each primary stage. Each growth stage consists of a primary and secondary stage and has both a mnemonic code and numerical index associated with it. The codes were designed to be easily memorized and are useful for applying the system in the field. The numerical index is included so that the stages can be expressed quantitatively.

Lignin is known to impede conversion of lignocellulose into ethanol. In this study, forage sorghum plants carrying brown midrib (bmr) mutations, which reduce lignin contents, were evaluated as bioenergy feedstocks. The near-isogenic lines evaluated were: wild type, bmr-6, bmr-12, and bmr-6 bmr-12 double mutant. The bmr-6 and bmr-12 mutations were equally efficient at reducing lignin contents (by 13% and 15%, respectively), and the effects were additive (27%) for the double mutant. Reducing lignin content was highly beneficial for improving biomass conversion yields. Sorghum biomass samples were pretreated with dilute acid and recovered solids washed and hydrolyzed with cellulase to liberate glucose. Glucose yields for the sorghum biomass were improved by 27%, 23%, and 34% for bmr-6, bmr-12, and the double mutant, respectively, compared to wild type. Sorghum biomass was also pretreated with dilute acid followed by co-treatment with cellulases and Saccharomyces cerevisiae for simultaneous saccharification and fermentation (SSF) into ethanol. Conversion of cellulose to ethanol for diluteacid pretreated sorghum biomass was improved by 22%, 21%, and 43% for bmr-6, bmr-12, and the double mutant compared to wild type, respectively. Electron microscopy of dilute-acid treated samples showed an increased number of lignin globules in double-mutant tissues as compared to the wild-type, suggesting the lignin had become more pliable. The mutations were also effective for improving ethanol yields when the (degrained) sorghum was pretreated with dilute alkali instead of dilute acid. Following pretreatment with dilute ammonium hydroxide and SSF, ethanol conversion yields were 116 and 130 mg ethanol/g dry biomass for the double-mutant samples and 98 and 113 mg/g for the wild-type samples.

A new method of determining in vitro dry matter digestibility (IVDMD) was recently developed in which the digestion is conducted with the forage samples in filter bags. Our objective was to compare the filter bag and conventional IVDMD analysis methods using smooth bromegrass (Bromus inermis Leyss.), switchgrass (Panicum virgatum L.), and forage sorghum [Sorghum bicolor (L.) Moench] samples. In addition, the filter bag analysis systems for determining non‐sequential neutral and acid detergent fiber (NDF and ADF), respectively, were compared with the non‐sequential conventional analysis systems. In the filter bag systems, the forage samples are sealed in filter bags and the analyses are conducted on a batch basis rather than on an individual basis as in the conventional IVDMD and fiber analysis procedures. The filter bag analysis methods produced results similar to the conventional methods and ranked the forage samples in the same relative order.

brown midrib6 (bmr6) affects phenylpropanoid metabolism, resulting in reduced lignin concentrations and altered lignin composition in sorghum (Sorghum bicolor). Recently, bmr6 plants were shown to have limited cinnamyl alcohol dehydrogenase activity (CAD; EC 1.1.1.195), the enzyme that catalyzes the conversion of hydroxycinnamoyl aldehydes (monolignals) to monolignols. A candidate gene approach was taken to identify Bmr6. Two CAD genes (Sb02g024190 and Sb04g005950) were identified in the sorghum genome based on similarity to known CAD genes and through DNA sequencing a nonsense mutation was discovered in Sb04g005950 that results in a truncated protein lacking the NADPH-binding and C-terminal catalytic domains. Immunoblotting confirmed that the Bmr6 protein was absent in protein extracts from bmr6 plants. Phylogenetic analysis indicated that Bmr6 is a member of an evolutionarily conserved group of CAD proteins, which function in lignin biosynthesis. In addition, Bmr6 is distinct from the other CAD-like proteins in sorghum, including SbCAD4 (Sb02g024190). Although both Bmr6 and SbCAD4 are expressed in sorghum internodes, an examination of enzymatic activity of recombinant Bmr6 and SbCAD4 showed that Bmr6 had 1 to 2 orders of magnitude greater activity for monolignol substrates. Modeling of Bmr6 and SbCAD4 protein structures showed differences in the amino acid composition of the active site that could explain the difference in enzyme activity. These differences include His-57, which is unique to Bmr6 and other grass CADs. In summary, Bmr6 encodes the major CAD protein involved in lignin synthesis in sorghum, and the bmr6 mutant is a null allele.

Brown midrib mutations and their importance to the utilization of maize, sorghum, and pearl millet lignocellulosic tissues" (2010). Papers in Plant Pathology. 453.

Total mixed rations containing conventional forage sorghum, brown midrib (bmr)-6 forage sorghum, bmr-18 forage sorghum, or corn silage were fed to Holstein dairy cows to determine the effect on lactation, ruminal fermentation, and total tract nutrient digestion. Sixteen multiparous cows (4 ruminally fistulated; 124 d in milk) were assigned to 1 of 4 diets in a replicated Latin square design with 4-wk periods (21-d adaptation and 7 d of collection). Diets consisted of 40% test silage, 10% alfalfa silage, and 50% concentrate mix (dry basis). Acid detergent lignin concentration was reduced by 21 and 13%, respectively, for the bmr-6 and bmr-18 sorghum silages when compared with the conventional sorghum. Dry matter intake was not affected by diet. Production of 4% fat-corrected milk was greatest for cows fed bmr-6 (33.7 kg/d) and corn silage (33.3 kg/d), was least for cows fed the conventional sorghum (29.1 kg/d), and was intermediate for cows fed the bmr-18 sorghum (31.2 kg/ d), which did not differ from any other diet. Total tract neutral detergent fiber (NDF) digestibility was greatest for the bmr-6 sorghum (54.4%) and corn silage (54.1%) diets and was lower for the conventional (40.8%) and bmr-18 sorghum (47.9%) diets. In situ extent of NDF digestion was greatest for the bmr-6 sorghum (76.4%) and corn silage (79.0%) diets, least for the conventional sorghum diet (70.4%), and intermediate for the bmr-18 sorghum silage diet (73.1%), which was not different from the other diets. Results of this study indicate that the bmr-6 sorghum hybrid outperformed the conventional sorghum hybrid; the bmr-18 sorghum was intermediate between conventional and bmr-6 in most cases. Additionally, the bmr-6 hybrid resulted in lactational

Lignin content of crop plants has been reduced by traditional plant breeding, natural and induced mutations, and insertion of transgenes. The effects of these genes and associated lower lignin content have been examined in terms of agricultural fitness or with regard to economically harvestable yields of useful plant products, or, in the case of some perennial species, survivability over multiple years. In general, crop yields are depressed by significant reductions in lignin content. Other negative effects observed in plants with lowered lignin contents include lodging and reduction of long‐term survival of some perennial species. However, the interactions of genes involved in lignin metabolism with genetic background and the environment in which the low‐lignin crop is cultivated are substantial. Examples are provided that demonstrate that lignin can be reduced in specific lines or populations without damaging fitness. It is concluded that it will be essential to incorporate lignin reducing genes into numerous genetic backgrounds and combinations, and evaluate the resulting lines in diverse environments, to discover optimal combinations and to obtain a true measure of value and fitness in agricultural systems.

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