O k l a h o m a N S F E P S C o R B i o e n e r g y R e s e a r c h a n d E d u c a t i o n STUDENT #1 SYNCHRONIZATION AND ISOLATION OF SWITCHGRASS FOR INTERECOTYPIC HYBRID DEVELOPMENT Laxman Adhikari and Yanqi Wu Department of Plant and Soil Sciences, Oklahoma State University
Objectives To know the flowering behavior of upland and lowland switchgrass. To determine the effects of synchronization and isolation in interecotypic hybrid development. To identify male sterile genotypes in interecotypic hybrid.
Materials and Methods Earlier flowering upland plants 1. were trimmed. Crossing pairs were isolated. 2. a b Hybrid seeds were collected. 3. Hybrid seeds’ genetic origins were 4. identified using SSR markers. Male sterile lines were identified c 5. d using pollen stainability and pollen Fig 1. (a) Isolated synchronized upland and lowland germination . plants, (b) Panicles from two ecotypes crossing each other, (c) hybrid seeds, and (d) hybrid plants.
Results Hybrid % Selfed % Contaminated % Hybrid % Selfed % Contaminated % A B 100 100 90 90 80 80 70 70 Hybrid % 60 Hybrid % 60 50 50 40 40 30 30 20 20 10 10 0 0 RC1 RC2 RC3 RC4 RC6 RC7 RC8 C1 C2 C3 C4 C5 C6 C7 C8 C9 Cross ID (upland progenies) Reciprocal cross ID (lowland progenies) Fig 2. (A) F1 upland progenies and their genetic origin, and (B) lowland F1 progenies and their genetic origin.
Conclusions Proper synchronization of reciprocal parents yielded 58 to 100 % interecotypic hybrids. Improper synchronization yielded relatively higher selfed seeds of reciprocal parents. The F1 genotypes C-8-17 and RC-3-3 were detected as possible male sterile line, no pollen germination. The information from this study will be valuable in the development of hybrid switchgrass.
O k l a h o m a N S F E P S C o R B i o e n e r g y R e s e a r c h a n d E d u c a t i o n STUDENT #2 An equilibrium based process modeling of a packed bed scrubbing system for the removal of model tar compounds Prakash Bhoi, Research Engineer Dr. Krushna Patil, Assistant Researcher Dr. Ajay Kumar, Assistant Professor Dr. Raymond Huhnke, Professor Biosystems & Agricultural Engineering Department Oklahoma State University, Stillwater, OK 74078
Objectives To develop an equilibrium based process model of a wet packed bed scrubber for the removal of model tar compounds. Equation of state (EOS) models Activity coefficient models To study the effect of important variables on the removal efficiency of model tar compounds Packing bed height Solvent temperature Liquid-to-gas (L/G) ratio
Methods Thermodynamic property methods: Model Oil in Peng-Robinson syngas RK-Soave Solvent: Soybean oil Packing media: 6 mm Raschig ring Raschig Ring Values RadFrac Characteristics Density, Kg/m 3 900 Surface are, m 2 /m 3 900 Packing factor, 1/m 2300 Model Oil out Void fraction, % 89 syngas
Results Peng-Robinson 100 Tar Removal Efficiency, % 80 L/G = 0.52 60 L/G = 0.45 40 L/G = 0.37 20 0 20 30 40 50 60 70 Solvent Temperature, °C
Conclusions Both property models (Peng-Robinson and RK-Soave) lead to comparable results. Packed bed height significantly increases tar removal efficiency. Solvent temperatures above 40°C significantly reduce tar removal efficiency. An increase in liquid-to-gas (L/G) ratio substantially increases tar removal efficiencies for solvent temperatures above 40°C.
O k l a h o m a N S F E P S C o R B i o e n e r g y R e s e a r c h a n d E d u c a t i o n STUDENT #3 Genome Sequence of the Anaerobic Gut Fungi Orpinomyces sp. strain C1A MB Couger, Noha H. Youssef, Audra S. Liggenstoffer, and Mostafa Elshahed Oklahoma State University Stillwater, Oklahoma.
Objectives Establish a high quality, well annotated, genome sequence from a member of the anaerobic fungal genera Neocallamastix Identify the unique salient features of the genome and conduct comparative analysis to other microbial genomes Identify enzymatic components of the genome that allows it to have the ability to thrive in the Rumen.
Methods 10x 100x Coverage Coverage 35GB 22,000 Models 100MB 3.5KB N50 1080 n50 Final Gene Models 16,347 Models Average Gene Length 1.6KB
Results Genome size 100.95 MB Number of Contigs 32,574 Protein Coding 20.60% Non- coding intergenic 73.60% Non-coding introns 5.10% rRNA 0.67% 5.8S 183 (30,763 bp) 18S 272 (168,110 bp) 28S 366 (457,301 bp) tRNA 0.06% 770 (58,292 bp) Number of Genes 16,347 Number of Genes with transcripts 14,009 -60 -40 -20 0 20 Bacterial P_anse 0.4 Average Gene Length 1623 A_oryz GH61 M_gris Homolog 247 M_ther 35,697 Number of Intron GH16GH18 T_rees 20 0.2 GH3 N_cras P_chry GH47 P_plac GH13 GH28 Introns/gene 2.18 GH43 GH2 PC2 GH31 R_oryz 0.0 GH5 Rumen GH1 0 GH6 Average Intronlength 163 GH8 GH45 A_mac GH11 GH26 S_punc GH48 M_circ GH10 B_dend Homolog 141 C_phyt GH9 -0.2 C_obs GC content 17.00% F_succ A_ther -20 R_alba Orpinomyces C_ther Protein Coding 26.80% -0.4 Eukaryotic -0.8 -0.6 -0.4 -0.2 0.0 0.2 0.4 Intergenic 14.80% PC1 Homolog 110 Intron 8.10% SSR Repeats 4.90% TE repeats 3.31%
Conclusions Analysis of the Genome of Orpinomyces C1A reveals a distinct genome structure from other members of Mycota. Anaerobic fungi contain a uniquely evolved enzymatic system for plant cell wall degradation, many members of which where obtained from horizontal gene transfer from other prokaryotic members residing in the rumen. C1A contains the capacity to degrade all major chemical moieties found in hemicellulose. This unique system combined with the invasiveness of fungi make this organism a very promising agent for consolidated bioprocessing.
O k l a h o m a N S F E P S C o R B i o e n e r g y R e s e a r c h a n d E d u c a t i o n STUDENT #4 Alkylation Reactions for the Upgrading of Bio-oil in the Presence of Liquid Water Using Hydrophobic Zeolites Miguel A. Gonzalez Borja, Daniel E. Resasco School of Chemical, Biological & Materials Engineering University of Oklahoma
Objectives To develop bio-oil upgrading strategies that maximize the yield of liquid products. To evaluate the performance of water-resistance catalyst for alkylation reactions in aqueous media. To understand the reaction pathways for the alkylation of phenolic compounds with 2-isopropanol.
Methods ALKYLATION MULTI-STAGE PYROLYSIS CHALLENGES • Bio-oil unstability upon heating FRACTION I Light Oxygenates • Phase separation 2-Propanol • Deactivation of catalyst by water T increase FRACTION II APPROACH Sugar derived • Work in liquid phase • Use catalyst that remains at FRACTION III liquid-liquid interphase Phenolics • Use catalyst that is stable in the m-Cresol presence of water. HYDROPHOBIC ZEOLITE IN A BIPHASIC LIQUID PHASE REACTOR
Results HYDROPHOBIC ZEOLITE PERFORMANCE 0.6 Regular Zeolite / Monophasic m-Cresol Conversion (mol/l) Hydrophobic Zeolite / Monophasic 0.5 Regular Zeolite / Biphasic Hydrophobic Zeolite / Biphasic 0.4 0.3 0.2 0.1 0.0 0 5 10 15 20 25 Time (hours)
Conclusions Alkylation reactions between light oxygenates and phenolics appear to be an effective strategy for bio-oil upgrading while maximizing the yield of liquid products. Hydrophobic zeolites that remain at the oil-water interphase posses improved stability for alkylation reactions in the presence of liquid water. 2-Propanol can be incorporated into the aromatic ring of phenolics via alkylation or via etherification. Ethers can in turn convert to the alkylated product via trans-alkylation.
O k l a h o m a N S F E P S C o R B i o e n e r g y R e s e a r c h a n d E d u c a t i o n STUDENT #5 Identification of Grass Cell Wall Synthesis Genes by Correlation Analysis between Gene Expression and Cell Wall Composition F . L i n a , C . M a n i s s e r i b , A . F a g e r s t r o m d , B . W i l l i a m s c , D . M . C h i n i q u y b , c , M . L . P e c k a , P . S a h a a , M . V e g a - S a n c h e z b , c , J . U . F a n g e l d , W . T . W i l l a t s d , H . V . S c h e l l e r b , P . C . R o n a l d b , c , L . E . B a r t l e y a , b , c a D e p a r t m e n t o f M i c r o b i o l o g y a n d P l a n t B i o l o g y , U n i v e r s i t y o f O k l a h o m a , N o r m a n , O K 7 3 0 1 9 b J o i n t B i o E n e r g y I n s t i t u t e , E m e r y v i l l e , C A 9 4 6 0 8 a n d L a w r e n c e B e r k e l e y N a t i o n a l L a b o r a t o r y , B e r k e l e y , C A 9 4 7 2 0 c D e p a r t m e n t o f P l a n t P a t h o l o g y a n d T h e G e n o m e C e n t e r , U n i v e r s i t y o f C a l i f o r n i a , D a v i s , C A 9 5 6 1 6 d D e p a r t m e n t o f P l a n t a n d E n v i r o n m e n t a l S c i e n c e s , U n i v e r s i t y o f C o p e n h a g e n , D e n m a r k
Objectives Focus on grass-specific cell wall biosynthesis. Develop a correlation based method to identify cell wall synthesis genes Improve grass cell walls as a feedstock for biofuel production
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