Progress 02/06/08 to 02/05/13
Outputs
Progress Report Objectives (from AD-416): The long-term objectives of this project are the development of sorghum (Sorghum bicolor) germplasm lines with improved bioenergy, feed, and food value, and the elucidation of genetic, biochemical, and biological factors impacting these characters. Over the next five years, the following specific objectives will be addressed: 1) Identify and evaluate genes to improve sorghum for bioenergy, food, and feed traits, 2) Develop a better understanding of genes and fundamental mechanisms controlling cell wall formation and energy availability, and 3) Develop molecular and other technologies for monitoring sorghum fungal pathogens and determine the effects of sorghum genetic modification for bioenergy on pathogen populations. Approach (from AD-416): The project utilizes a diverse set of technologies and approaches that are best delineated by objective: Objective 1 will be addressed primarily using traditional whole-plant plant breeding techniques, animal feeding trials, and established laboratory methods for assessment of feed and bioenergy value. Objective 2 will be addressed using current molecular and biochemistry technologies including PCR, RT-PCR, HPLC, microarrays, GC-MS. Objective 3 incorporates both field-based experiments and current molecular biology technologies. All experiments will utilize sound statistical designs to allow hypothesis testing at established levels of probability. This five-year project ended February of 2013, and was renewed as project number 5440-21220-032-00D, Genetic Improvement of Sorghum for Non-Grain Energy Uses. The major accomplishments of this project over the past 5 years are summarized here. Sorghum elite lines were bred that incorporated brown midrib (bmr) 6 and 12 mutations to reduce the concentration of the cell wall polymer lignin. These lines were publicly released, which has served as the basis for full range of scientific research on bmr6 and bmr12 from basic cell wall biochemistry to new agricultural applications. The genes encoding three bmr mutant loci were cloned, characterized and discovered to encode enzymes in lignin synthesis. These studies linked phenotypes, mutations, protein levels and enzymatic activity together to provide a complete picture of these bmr lines. This information allows plant breeders to develop strategies to modify lignin content and its composition in sorghum, which also has implications for lignin modification in other bioenergy grasses. Whether bmr6 and bmr12 affected the susceptibility of sorghum to fungal pathogens was evaluated, because lignin accumulation is a common response to pathogens. These studies indicated that using bmr6 and bmr12 to enhance sorghum for bioenergy and livestock feed does not increase disease susceptibility and, for some pathogens, results in increased resistance. Near-isogenic lines containing the waxy mutation were developed for the northern sorghum-growing region of the USA. These lines were shown to have greater digestibility of the grain for livestock feed and conversion efficiency to ethanol. These lines have provided a basis to biochemically characterize Waxy, the sorghum granule-bound starch synthase and its function in amylose synthesis. waxy lines are valuable germplasm for enhancing sorghum grain for conversion to bioenergy, for food processing and for use in research. Grain from waxy and normal sorghum lines was screened for fungal pathogens, which can affect grain yield or grain quality. The results showed that waxy sorghum lines were not more susceptible to grain infections than normal sorghum. Biological controls are important management tools for plant pests and pathogens. Ten soil bacteria isolates were identified which have potential as biological control agents, because they inhibit the growth of soil borne fungal pathogens that infect both sorghum grain and stalks. This information may lead to new ways to control these pathogens, which impact both sorghum grain and biomass yields. Accomplishments 01 Release of elite brown midrib (bmr6 and bmr12) sorghum lines. Since the first release of elite bmr lines in 2005, over 1200 seed packets from these lines have been distributed without charge and without restrictions on their use, to private companies, universities, and national labs on six continents. These lines are major sources of bmr6 and bmr12 germplasm for the global sorghum community. 02 Demonstrated that food-quality white grain sorghum lines are not more susceptible to diseases. White sorghum grain grown on unpigmented (tan) plants is highly desirable for food and feed uses in contrast to the same white grain grown on pigmented (purple) plants. However, plant pigment has been suggested to protect plants against two fungal diseases: grain mold and head smut. White grain from purple and tan plants was collected from three locations and screened for grain mold infection. The results showed no indication that grain from tan or purple plants had greater levels of infection. In addition, these plants were evaluated for head smut, a common sorghum disease in Corpus Christi, TX. The purple plants were more susceptible to this disease than tan plants at this location. These results indicate that sorghum grain grown on tan plants will not result in greater grain mold and head smut disease incidences, which is valuable information for the development of food-quality grain sorghum. 03 Evaluation of beef cattle grazed on bmr and conventional grain sorghum residue. Brown midrib 12 (bmr12) mutant lines have reduced lignin content in their tissue as compared to conventional plants. Following grain harvest, cattle were grazed on bmr12 and conventional grain sorghum residue, and animal weight gain was monitored. There was a significant (34%) increased in the weight gain of cattle grazing on bmr12 residue relative to cattle grazing on conventional sorghum residue. Therefore, incorporating the bmr trait into grain sorghum increases the value of the residue as a livestock feed.
Impacts
(N/A)
Publications
- Funnell-Harris, D.L., Prom, L.K., Pedersen, J.F. 2013. Isolation and characterization of the grain mold fungi, Cochliobolus and Alternaria spp., from sorghum using semi-selective media and DNA sequence analyses. Canadian Journal of Microbiology. 59(2): 87-96.
- Pedersen, J.F., Sattler, S.E., Anderson, W.F. 2012. Evaluation of public sweet sorghum A-lines for use in hybrid production. BioEnergy Research. 6: 91-102. DOI:10.1007/s12155-012-9231-1
- Sattler, S.E., Toy, J.J., Aketch Okeno, J., Funnell-Harris, D.L., Pedersen, J.F. 2013. Registration of N614, A3N615, N616, and N617 Shattercane Genetic Stocks with cytoplasmic or nuclear male-sterility and juicy or dry midribs. Journal of Plant Registrations. 7: 245-249. DOI 10.3198/jpr201209. 0033crgs
- Funnell-Harris, D.L., Sattler, S.E., Pedersen, J.F. 2013. Characterization of fluorescent Pseudomonas spp. associated with roots and soil of two sorghum genotypes. European Journal of Plant Pathology. 136 (3): 469-481.
- Sattler, S.E., Funnell-Harris, D.L. 2013. Modifying lignin to improve bioenergy feedstocks: strengthening the barrier against pathogens? Frontiers in Plant Science. 4: 70. DOI 10.3389/fpls.2013.00070
- Funnell-Harris, D.L., Prom, L.K., Sattler, S.E., Pedersen, J.F. 2013. Response of near isogenic sorghum lines, differing at the P locus for plant color, to grain mold and head smut fungi. Annals of Applied Biology. 163:91-101.
- Sukumaran, S., Xiang, W., Bean, S., Pedersen, J.F., Tuinstra, M.R., Tesso, T.T., Hamblin, M.T. and Yu, J. 2012. Association mapping for grain quality in a diverse sorghum collection. The Plant Genome. 5:126-135.
- Walker, A.M., Hayes, R.P., Youn, B., Vermerris, W., Sattler, S.E., Kang, C. 2013. Elucidation of the structure and reaction mechanism of Sorghum bicolor hydroxycinnamoyltransferase and its structural relationship to other CoA-dependent transferases and synthases. Plant Physiology. 162:640- 651. DOI
- Schmidt, J.J., Pedersen, J.F., Bernards, M.L., Lindquist, J.L. 2013. Rate of shattercane x sorghum hybridization in situ. Crop Science. 53:1677-1685.
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Progress 10/01/11 to 09/30/12
Outputs
Progress Report Objectives (from AD-416): The long-term objectives of this project are the development of sorghum (Sorghum bicolor) germplasm lines with improved bioenergy, feed, and food value, and the elucidation of genetic, biochemical, and biological factors impacting these characters. Over the next five years, the following specific objectives will be addressed: 1) Identify and evaluate genes to improve sorghum for bioenergy, food, and feed traits, 2) Develop a better understanding of genes and fundamental mechanisms controlling cell wall formation and energy availability, and 3) Develop molecular and other technologies for monitoring sorghum fungal pathogens and determine the effects of sorghum genetic modification for bioenergy on pathogen populations. Approach (from AD-416): The project utilizes a diverse set of technologies and approaches that are best delineated by objective: Objective 1 will be addressed primarily using traditional whole-plant plant breeding techniques, animal feeding trials, and established laboratory methods for assessment of feed and bioenergy value. Objective 2 will be addressed using current molecular and biochemistry technologies including PCR, RT-PCR, HPLC, microarrays, GC-MS. Objective 3 incorporates both field-based experiments and current molecular biology technologies. All experiments will utilize sound statistical designs to allow hypothesis testing at established levels of probability. Sorghum brown midrib (bmr) mutants have been developed which reduce lignin content of biomass, thereby increasing digestibility for livestock and conversion potential for cellulosic ethanol. New brown midrib mutants have been identified from an ARS population at Lubbock, TX and a series of these bmr12 mutants were characterized. These new mutant lines have the potential to provide additional genetic resources to modulate lignin content and composition. Lignin, has been previously implicated in helping plants defend themselves from pathogens. These new bmr lines were screened for susceptibility to stalk rot pathogens, which can reduce yield and contribute to lodging. None of the bmr lines showed increased susceptibility to the pathogen compared to non-modified or wild-type sorghum, and some bmr lines showed increased resistance to the pathogens. These results indicate that genetically reducing lignin concentration in sorghum using bmr mutants for bioenergy feedstock improvement should not increase susceptibility of sorghum to stalk rot pathogens. (301/03/A) (303/03/B) The waxy mutation reduces amylose starch and increases amylopectin starch concentration in sorghum, and together these changes in starch composition increase the digestibility of sorghum grain for livestock and the conversion efficiency of sorghum grain to ethanol. Near-isogenic lines containing the waxy mutation were developed that are adapted to the northern sorghum region of the USA. Grain from waxy and wild-type lines was screened for fungal pathogen, which can affect grain yield or affect grain quality. The results showed that waxy sorghum lines were not more susceptible to grain infections than wild-type lines. (301/03/C) (303/03/B) Biological controls are important management tools for plant pests and pathogens. Two soil bacteria were identified which have potential as biological control agents, because they inhibit the growth of soil borne fungal pathogens that infect both sorghum grain and stalks. This information may lead to new ways to control these pathogens, which impact both grain and biomass yields. (303/04/A) Accomplishments 01 Identification of Brown midrib 2 (Bmr2) gene. Lignin is the principal component that makes cell walls resistant to breakdown either in livesto digestive systems or in the cellulosic bioenergy conversion process. Brown midrib 2 (bmr2) is a mutant that reduces lignin content and alters lignin composition of sorghum cell walls. The mutation causing bmr2 was identified by scientists at University of Florida and ARS scientists at Lincoln, NE and shown to affect an enzyme involved in lignin synthesis. The function of the enzyme was characterized and determined to be largel responsible for an initial step in lignin synthesis. The bmr2 gene represents a strategic tool that can be used to improve sorghum biomass for bioenergy and forage and potentially could be used in other species. 02 Identification of bacteria with potential for biological control against sorghum pathogens. Fluorescent Pseudomonas bacteria were collected from roots and surrounding soil of different sorghum varieties and a wheat variety grown in two different soil types. These bacteria are able to produce compounds including an antibiotic that inhibit fungi, which caus diseases in sorghum. Two of bacteria isolated from sorghum roots inhibit the growth of five disease-causing fungi. These two bacteria have potential as biological agents to control disease-causing fungi in sorgh
Impacts
(N/A)
Publications
- Yan, S., Wu, X., Bean, S., Pedersen, J.F., Tesso, T., Chen, Y.R. and Wang, D. 2011. Evaluation of waxy grain sorghum for ethanol production. Cereal Chemistry. 88(6):589-595.
- Griess, J., Mason, S., Jackson, D., Galusha, T., Pedersen, J.F., Yaseen, M. 2011. Environment and hybrid influences on rapid visco analysis starch properties of food-grade grain sorghum. Crop Science. 51(4):1757-1766.
- Saballos, A., Sattler, S.E., Sanchez, E., Foster, T., Xin, Z., Kang, C., Pedersen, J.F., Vermerris, W. 2012. Sorghum Brown midrib 2 (Bmr2) gene encodes the major 4-coumarate Coenzyme A ligase involved in lignin synthesis. Plant Journal. 70:818-830.
- Lee, S.J., Warnick, T.A., Pattathill, S., Alveo-Maurosa, J.G., Serapiglia, M.J., Young, N.F., Schnell, D.J., Smart, L.B., Hahn, M.G., Pedersen, J.F., Mccormick Ford, H., Brown, V. 2012. Biological conversion assay for determining plant feedstock quality. Biotechnology for Biofuels. 5:5.
- Sattler, S.E., Palmer, N.A., Saballos, A., Greene, A.M., Xin, Z., Sarath, G., Vermerris, W., Pedersen, J.F. 2012. Identification and characterization of 4 missense mutations in brown midrib 12 (Bmr12); the caffeic O-methyltranferase (COMT) of sorghum. BioEnergy Research. 5: 855- 865. DOI 10.1007/s12155-012-9197.
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Progress 10/01/10 to 09/30/11
Outputs
Progress Report Objectives (from AD-416) The long-term objectives of this project are the development of sorghum (Sorghum bicolor) germplasm lines with improved bioenergy, feed, and food value, and the elucidation of genetic, biochemical, and biological factors impacting these characters. Over the next five years, the following specific objectives will be addressed: 1) Identify and evaluate genes to improve sorghum for bioenergy, food, and feed traits, 2) Develop a better understanding of genes and fundamental mechanisms controlling cell wall formation and energy availability, and 3) Develop molecular and other technologies for monitoring sorghum fungal pathogens and determine the effects of sorghum genetic modification for bioenergy on pathogen populations. Approach (from AD-416) The project utilizes a diverse set of technologies and approaches that are best delineated by objective: Objective 1 will be addressed primarily using traditional whole-plant plant breeding techniques, animal feeding trials, and established laboratory methods for assessment of feed and bioenergy value. Objective 2 will be addressed using current molecular and biochemistry technologies including PCR, RT-PCR, HPLC, microarrays, GC-MS. Objective 3 incorporates both field-based experiments and current molecular biology technologies. All experiments will utilize sound statistical designs to allow hypothesis testing at established levels of probability. Sub-objective 1a. Articles describing the chemistry and bioenergy potential of commercially deployed brown midrib genes bmr6 and bmr12 were previously published. Articles describing animal performance are being prepared. New brown midrib mutants have been identified at new loci in a mutant population from USDA ARS Lubbock. Allelism tests, agronomic trials, and fiber chemistry have been completed. Data analyses and summary is underway. Sub-objective 1b. Waxy lines have been provided to collaborators at Kansas State, UC Berkeley, and the Agricultural Research Institute for South-East Region of Russia. A manuscript was published on the contributions of protein and starch chemistry on digestibility, and another submitted describing the effects of waxy sorghum on ethanol production. Field trials have been completed on a set of waxy/wild-type isolines, R-lines, A/B-lines, and another set of isolines with wxa and wxb alleles. Sub-objective 1c. BC1S2 families were planted to the field in spring 2011. Selections will be made at the end of the current growing season. Sub objective 2a. Assessment of bmr-6 gene expression was completed, and analysis of genomic data is ongoing. Sub objective 2b. A sorghum nuclear extract protocol was not developed because promoter elements were not conserved within lignin biosynthetic genes. An alternative strategy was developed using a putative lignin regulatory factor SbMYB68, selected based on comparative genomics. This factor is under investigation. Sub-objective 3a. Procedures for molecular analyses of Fusarium, Alternaria, and Curvularia spp. were established. A manuscript is in preparation for characterization of Alternaria, and Curvularia spp. A procedure for collection of fungi from air was established, and a manuscript for Fusarium spp. published. One-hundred forty four sequences were submitted to GenBank. Sub-objective 3b. Near isogenic plant color line analyses were completed and the resulting manuscript submitted. Screening grain from waxy accessions for fungal colonization, and fungal identifications, approx one-third completed. �Clean� grain for in vitro grain infections assays were produced in the greenhouse. Sub-objective 3c. Stalk rot analyses of bmr near isolines (bmr6, bmr12, bmr6 brm12 double mutants) in two genetic backgrounds using pathogens, Fusarium thapsinum and F. verticillioides, were completed. A similar study using F. proliferatum is underway. Assays to establish conditions for inoculation by Macrophomina phaseolina have been completed and stalk rot analyses with the near isolines are underway. Sub-objective 3d. RFLP analysis of 24 antibiotic producing fluorescent Pseudomonas spp. nearly complete. Inhibition assays with these isolates against five sorghum fungal pathogens completed. Accomplishments 01 Conversion of elite public sorghum strains to brown midrib leads to worl wide distribution and use. Mutant brown midrib strains of sorghum initially released to the public in 1977 had superior digestibility due reduced lignin content, but were not immediately useful to the seed industry or farmers due to associated detrimental effects such as increased lodging and reduced overall fitness. USDA-ARS researchers at Lincoln, NE, addressed these problems by incorporating two brown midrib genes, bmr-6 and bmr-12 into elite public lines using long-term targeted breeding and research. Research results have demonstrated the ability of hybrid vigor to overcome decreased yield, shown reduced disease incidenc to be associated with these genes, and have shown higher biomass ethanol conversion efficiencies to be associated with these genes. Released genetic materials have supported commercial efforts to develop markets f proprietary hybrids throughout the world. Research based on these materials has identified the biochemical and molecular genetic changes responsible for the differences in lignin chemistry leading to the identification of targets for improvement of other bioenergy crops.
Impacts
(N/A)
Publications
- Sahoo, L., Schmidt, J.J., Pedersen, J.F., Lee, D.L., Lindquist, J.L. 2010. Growth and fitness components of wild X cultivated Sorghum bicolor (Poaceae) hybrids in Nebraska. American Journal of Botany. 97(10): 1610- 1617.
- Wong, J.H., Marx, D.B., Wilson, J.D., Lemaux, P.G., Buchanan, B.B., Pedersen, J.F. 2010. Principal component analysis and biochemical characterization of protein and starch reveal primary targets for improving sorghum grain. Plant Science. Volume 179 (2010) 598-611.
- Funnell-Harris, D.L., Pedersen, J.F. 2011. Presence of Fusarium spp. in air and soil associated with sorghum fields. Plant Disease 95: 648-656.
- Goff, B., Moore, K., Fales, S., Nikolau, B., Pedersen, J.F. 2011. Comparison of the Use of Gas Chromotography, Spectrophotometry, and Near Infrared Spectropscopy to Quantify Prussic Acid Potential in Forages. Journal of Agriculture and Food Chemistry. 1523-1526.
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Progress 10/01/09 to 09/30/10
Outputs
Progress Report Objectives (from AD-416) The long-term objectives of this project are the development of sorghum (Sorghum bicolor) germplasm lines with improved bioenergy, feed, and food value, and the elucidation of genetic, biochemical, and biological factors impacting these characters. Over the next five years, the following specific objectives will be addressed: 1) Identify and evaluate genes to improve sorghum for bioenergy, food, and feed traits, 2) Develop a better understanding of genes and fundamental mechanisms controlling cell wall formation and energy availability, and 3) Develop molecular and other technologies for monitoring sorghum fungal pathogens and determine the effects of sorghum genetic modification for bioenergy on pathogen populations. Approach (from AD-416) The project utilizes a diverse set of technologies and approaches that are best delineated by objective: Objective 1 will be addressed primarily using traditional whole-plant plant breeding techniques, animal feeding trials, and established laboratory methods for assessment of feed and bioenergy value. Objective 2 will be addressed using current molecular and biochemistry technologies including PCR, RT-PCR, HPLC, microarrays, GC-MS. Objective 3 incorporates both field-based experiments and current molecular biology technologies. All experiments will utilize sound statistical designs to allow hypothesis testing at established levels of probability. 1a. Data describing the effect of bmr-12 on grain sorghum and steer performance was analyzed, and data on the effect of bmr-12 silage on dairy performance was summarized. An article describing the effect of stacked bmr genes on ETOH was published. 1b. Backcrossing to male-sterilize selected elite B-lines was continued and S4 lines still segregating for waxy alleles were identified. 1c. Backcrossing and selfing of high energy and high starch content introductions into elite backgrounds was continued. 2a. Expressed Bmr12 (COMT) in E. coli and made site-directed mutation for bmr12 alleles. In collaboration with the National Genome Resource Center, Santa Fe, NM, detected SNPs within transcripts from the stalk between wild-type and bmr mutant lines. 2b. Demonstrated that promoter elements were not conserved within lignin biosynthetic genes. 3a. Conducted assays to measure inhibition rates of sorghum grain pathogens grown on different semi-selective medium. Isolated crude preparations of a toxin from Curvularia species; established genotype specificity in production. Established three loci for genotyping of Fusarium spp. and have sequenced the three loci from over 200 isolates. Discovered a new genotype within the Fusarium incarnatum/F. equiseti species complex. Found six species of Alternaria spp. in grain grown in Nebraska and Texas, four of which have never been reported on sorghum, and Embellesia spp., which also has not been previously reported on sorghum. Paper describing Fusarium spp. epidemiology in bmr grain published. 3b. Screening of grain from near isogenic plant color lines for Alternaria, Fusarium and Curvularia spp. to be completed by end of FY2010. 3c. Established conditions for stalk rot inoculations by Fusarium verticillioides. Assay conditions for inoculation by Macrophomina phaseolina are being established and to be completed by end of FY2010. Three repetitions to screen wild-type, bmr6, bmr12 and bmr6, bmr12 double mutant lines in two genetic backgrounds for resistance to stalk rot by Fusarium thapsinum to be completed by end of FY2010. 3d. Molecular genetic characterization of select isolates with genes for antibiotic production continued. Paper describing microbial numbers and biochemical and genetic characterization of fluorescent Pseudomonas associated with sorghum in growth-chamber and field studies accepted for publication. Accomplishments 01 Singular and stacked brown midrib 6 and 12 modify lignocellulosic chemistry. Brown midrib genes were introduced into the grain sorghum hybrid AWheatland x RTx430 by ARS researchers at Lincoln, NE, to reduce lignin content and improve digestibility of stalk residues following gra harvest for use as a bioenergy or livestock feedstock. Impacts of the genes bmr6, bmr12 and the bmr6bmr12 �stacked� double mutant on sorghum biomass composition and whole-plant physiology were assessed in a two ye field study. The lignin content of the stacked stalk residue was further reduced compared to bmr6 or bmr12, but it had digestibility comparable t bmr12 stalk. Analysis of stalk carbohydrate composition indicated that bmr12 had modestly increased amounts of cellulose, glucoarabinoxylan and soluble sugars. Together these results indicate that bmr12 had broader effects on stalk composition than bmr6 or stacked hybrid, and this increase in carbohydrates in bmr12 is another potential positive effect for downstream applications. This body of work has demonstrated that sorghum brown midrib mutants have reduced lignin and increased biomass digestibility, while having a minor impact on plant fitness and yield in hybrid backgrounds. 02 Brown midrib sorghum is associated with reduced pathogens. Lignin biosynthesis mutants, bmr6 or bmr12, were incorporated into sorghum grai lines to reduce lignin content and increase cell wall digestibility. Traditionally it was assumed that reduction in lignin biosynthesis would cause plants to be more susceptible to diseases. However, ARS researcher at Lincoln, NE showed that for infection by some Fusarium fungi, the mutant lines were actually more resistant. There was a significant reduction in grain infections of field grown plants and stalk infections of plants inoculated in the greenhouse. In particular, one Fusarium species, commonly-found in wild-type grain, was not detected in bmr12 grain. This research provides evidence that sorghum modified for increas bioenergy use could also be more resistant to some pathogens.
Impacts
(N/A)
Publications
- Funnell-Harris, D.L., Pedersen, J.F., Sattler, S.E. 2010. Alteration in Lignin Biosynthesis Restricts Growth of Fusarium Species in Brown Midrib Sorghum. Phytopathology 100:671-681.
- Ellis, D.D., Campbell, K., Grotenhuis, J.A., Jenderek, M.M., Pedersen, J.F. 2010. Crop Registration: The Pathway to Public Access of Plant Genetic Materials to Build Crops for the Future. Crop Science. 50:1151-1160.
- Funnell-Harris, D.L., Pedersen, J.F., Sattler, S.E. 2010. Soil and Root Populations of Fluorescent Pseudomonas spp. Associated with Seedlings and Field-grown Plants are Affected by Sorghum Genotype. Plant and Soil Journal. Volume 335: 439-455.
- Griess, J., Mason, S., Jackson, D., Galusha, T., Yaseen, M., Pedersen, J.F. 2010. Environment and Hybrid Influences on Food-Grade Sorghum Grain Yield and Hardness. Crop Science. 50: 1480-1489.
- Preiffer, T.W., Bitzer, M.J., Toy, J.J., Pedersen, J.F. 2010. Heterosis in Sweet Sorghum and Selection of a New Sweet Sorghum Hybrid for Use in Syrup. Crop Science. doi: 10.2135/cropsci2009.09.0475.
- Sattler, S.E., Funnell-Harris, D.L., Pedersen, J.F. 2010. Brown Midrib Mutations and Their Importance to the Utilization of Grasses. Plant Science. 178:229-238
- Sattler, S.E., Funnell-Harris, D.L., Pedersen, J.F. 2010. Efficacy of Singular and Stacked Brown Midrib 6 and 12 in Modification of Lignocellulose and Grain Chemistry. Journal of Agricultural and Food Chemistry. 58:3611-3616.
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Progress 10/01/08 to 09/30/09
Outputs
Progress Report Objectives (from AD-416) The long-term objectives of this project are the development of sorghum (Sorghum bicolor) germplasm lines with improved bioenergy, feed, and food value, and the elucidation of genetic, biochemical, and biological factors impacting these characters. Over the next five years, the following specific objectives will be addressed: 1) Identify and evaluate genes to improve sorghum for bioenergy, food, and feed traits, 2) Develop a better understanding of genes and fundamental mechanisms controlling cell wall formation and energy availability, and 3) Develop molecular and other technologies for monitoring sorghum fungal pathogens and determine the effects of sorghum genetic modification for bioenergy on pathogen populations. Approach (from AD-416) The project utilizes a diverse set of technologies and approaches that are best delineated by objective: Objective 1 will be addressed primarily using traditional whole-plant plant breeding techniques, animal feeding trials, and established laboratory methods for assessment of feed and bioenergy value. Objective 2 will be addressed using current molecular and biochemistry technologies including PCR, RT-PCR, HPLC, microarrays, GC-MS. Objective 3 incorporates both field-based experiments and current molecular biology technologies. All experiments will utilize sound statistical designs to allow hypothesis testing at established levels of probability. Significant Activities that Support Special Target Populations 1.a. Manuscripts delineating the effects on bmr-6, bmr-12-and stacked bmr- 6+12 on chemical makeup, ethanol conversion, and dairy nutrition were submitted in collaboration with USDA-ARS Peoria and the WH Miner Institute. New brown midrib lines from USDA-ARS Lubbock were crossed to testers and classified for allelism with known bmr genes in sorghum. Brown midrib genes were identified that were not allelic to genes at known loci. 1.b. Yield trials of waxy B and R-lines were completed. One generation converting waxy B-lines into A3 male sterile cytoplasm has been completed, and nurseries to accomplish the second backcross are established in the field. The first year of a field trial comparing GBSS+ and GBSS- near isolines was planted and is in progress. 1.c. Backcrosses of high starch and high energy lines F1s with elite lines were completed. Selfing is underway. 2.a. Expressed the Bmr6 protein in E. coli and in planta to examine a unique enzymatic feature, histine 57. Expressed Bmr12 (COMT) in E. coli and developed an enzyme assay. In collaboration with the National Genome Resource Center, Santa Fe, NM, sequenced 192 million reads to characterize the difference in the stalk transcriptome between wild-type and bmr mutant lines. 2.b. Identified potential transcription factors which activate lignin biosynthesis based on amino acid sequence identity. 3.a. Established semi-selective media for screening for Curvularia spp. from grain. Established use of sequence analysis of translation elongation factor (TEF) gene to genotype Fusarium spp. Established methodologies for identifying Alternaria isolates to species. Substantial progress in screening near isogenic plant color lines for Alternaria, Fusarium and Curvularia spp. 3.b. Screened of grain from waxy accessions for infections by Alternaria, Fusarium and Curvularia. 3.c. Established stalk rot infection assay for Fusarium thapsinum. Have begun bioassays comparing brown midrib6 (bmr), bmr12, bmr6 and bmr12 double mutants and near isogenic RTx430 and Wheatland wild-type lines for response to stalk infection by F. thapsinum. Collaborated with Louis Prom to conduct field study of bmr6, bmr12, bmr6 and bmr12 double mutants and near isogenic RTx430 lines for infection by anthracnose (Colletotrichum sublineolum). 3.d. Growth chamber bioassays completed. Second paper describing microbial numbers and biochemical and genetic characterization of fluorescent Pseudomonas associated with sorghum submitted. Further molecular genetic characterization of select isolates with genes for antibiotic production on-going. Technology Transfer Number of New/Active MTAs(providing only): 2
Impacts
(N/A)
Publications
- Funnell-Harris, D.L., Pedersen, J.F. 2008. Inoculation strategies to assess biological interactions between fusarium and alternaria species infecting sorghum. Canadian Journal of Plant Pathology 30: 404-413.
- Pedersen, J.F., Toy, J.J., Funnell-Harris, D.L., Sattler, S.E., Oliver, A. L. 2008. Registration of BN611, A/BN612, RN613 Sorghum Genetic Stocks with Stacked bmr-6 and bmr-12 Genes. Journal of Plant Registrations 2:258-262.
- Wong, J.H., Lau, T., Cai, N., Singh, J., Pedersen, J.F., Vensel, W.H., Hurkman II, W.J., Wilson, J.D., Lemaux, P.G., Buchanan, B.B. 2008. Digestibility of Protein and Starch from Sorghum (Sorghum bicolor)is Linked to Biochemical and Structural Features of Grain Endosperm. Journal of Cereal Science. 49(1):73-82.
- Palmer, N.A., Sattler, S.E., Saathoff, A.J., Funnell-Harris, D.L., Pedersen, J.F., Sarath, G. 2008. Genetic background impacts soluble and cell wall-bound aromatics in brown midrib mutants of sorghum. Planta 229 (1): 115-127
- Sattler, S.E., Saathoff, A.J., Haas, E.J., Palmer, N.A., Funnell-Harris, D. L., Sarath, G., Pedersen, J.F. 2009. A nonsense mutation in a cinnamyl alcohol dehydrogenase gene is responsible for the sorghum brown midrib-6 phenotype. Plant Physiology 150 (2):584-95.
- Hooks, T., Marx, D.B., Kachman, S.D., Pedersen, J.F. 2009. Revista Columbiana de Estadstica. Columbian Journal of Statistics 32:17-31.
- Sattler, Scott E., Jaswinder Singh, Eric J. Haas, Lining Guo, Gautam Sarath, and Jeffrey F. Pedersen Two distinct waxy alleles impact the granule-bound starch synthase in sorghum. Molecular Breeding. Published online 6/5/09 DOI 10.1007/s11032-009-9296-5.
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Progress 10/01/07 to 09/30/08
Outputs
Progress Report Objectives (from AD-416) The long-term objectives of this project are the development of sorghum (Sorghum bicolor) germplasm lines with improved bioenergy, feed, and food value, and the elucidation of genetic, biochemical, and biological factors impacting these characters. Over the next five years, the following specific objectives will be addressed: 1) Identify and evaluate genes to improve sorghum for bioenergy, food, and feed traits, 2) Develop a better understanding of genes and fundamental mechanisms controlling cell wall formation and energy availability, and 3) Develop molecular and other technologies for monitoring sorghum fungal pathogens and determine the effects of sorghum genetic modification for bioenergy on pathogen populations. Approach (from AD-416) The project utilizes a diverse set of technologies and approaches that are best delineated by objective: Objective 1 will be addressed primarily using traditional whole-plant plant breeding techniques, animal feeding trials, and established laboratory methods for assessment of feed and bioenergy value. Objective 2 will be addressed using current molecular and biochemistry technologies including PCR, RT-PCR, HPLC, microarrays, GC-MS. Objective 3 incorporates both field-based experiments and current molecular biology technologies. All experiments will utilize sound statistical designs to allow hypothesis testing at established levels of probability. Significant Activities that Support Special Target Populations by CRIS Project Objectives: 1.a. Tissue samples of lines near-isogenic for bmr-6, bmr-12, and �stacked bmr-6+12� were sent to the W.S. Miner Institute for in compositional and situ diary feeding studies, and to USDA-ARS Peoria for fermentation studies. The 2nd year of a field-scale residue beef grazing study comparing isogenic wild type and bmr-12 grain hybrids is underway. An experiment comparing dairy performance when fed silage from wild-type and bmr-12 Atlas was completed, data analysis is pending. (NP301, Component 3) 1.b. New amylose-free (waxy) R-lines are undergoing their 2nd year of yield-trials. Crosses to verify fertility restoration reaction of new R- and B-line were made. Selected B-lines are being sterilized in A1 cytoplasm. Final selections of near-isogenic sister-lines pairs are being made in the field. Lines segregating for wxa and wxb alleles were identified and are being advanced. (NP301, Component 3) 1.c. Initial crosses of high starch content and high energy plant introduction lines with elite parents were accomplished using ms3 male sterility and selfing completed in winter greenhouse to recover male sterility expression. Back crosses are currently being made. (NP301, Component 3) 2.a. Identified the locus responsible for the bmr-6 phenotype, which contained a nonsense mutation in CAD80 gene. Expressed the CAD80 and CAD40 protein in E. coli and the enzymatic activity assay are being performed. Expressed BMR-12 (COMT) in E. coli and assays are pending. Plant tissue is being produce for a global gene expression study to be performed in collaboration with the National Genome Resource Center, Santa Fe, NM. (NP301, Component 3) 2.b. Four conserved non-coding sequences (CNS) in the CAD80 promoter shared with Brachypodium and Rice were identified. Conservation of CNS does not extend beyond the grasses. (NP301, Component 3) 3.a. Improved greenhouse assays were published. Field studies were expanded to Corpus Christi, TX to take advantage of higher pathogen pressure. (NP 303, Component 1) 3.b. Seed from low amylose (waxy) and wild-type lines has been collected. Lab screening to identify and isolate pathogens is scheduled to be initiated. (NP 303, Component 1) 3.c. Over 90 Fusarium spp. were isolated from grain and leaves of low lignin sorghum lines and wild-type counterparts. Sequencing of three genes from each fungal isolates has been completed. Greenhouse screens of low lignin sorghum lines inoculated with fungal sorghum isolates representing eight different species were conducted. (NP 303, Components 1, 3) 3.d Nearly 6000 antibiotic-producing bacteria were obtained from roots and soil of sorghum and wheat seedlings grown in field soil in growth chamber assays, and from roots and soil of field-grown sorghum. Biochemical and molecular genetic characterization of these isolates is complete. One growth chamber bioassay monitoring colonization of biocontrol bacterial isolates on sorghum roots and in soil was conducted. (NP 303, Component 4) Technology Transfer Number of New/Active MTAs(providing only): 2 Number of Other Technology Transfer: 1
Impacts
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Publications
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