Progress 10/01/07 to 09/30/13
Outputs
Target Audience: The outcome of this project was tailored to impact student learning and practicum, and benefit educators and farmers around Tuskegee University community - local, regional, state and international, and the global biotechnology, genomic, breeding, scientific and farmer communities at large. The biotechnology, genomic and breeding communities involved especially in peanut, sweetpotato, root and tuber, bioenergy, legume and vegetable research and production were targeted for two of the developed peanut genetic maps published and the sweetpotato storage root development work. The root and tuber crop communities may benefit from our yam procedures for germplasm conservation and improvement of other species, using limited laboratory resources. Changes/Problems: With the advent of renewable energy, the project sought to screen sweetpotato endogenous enzyme impact on ethanol production and also identify plants best suited for bioenergy production. Investigation of Moso Bamboo along with Miscanthus, Sugar Cane, Gamma Gras and 2 Sweetpotato (TUO2, DMO1-158-204) cultivars Genomic DNA were performed through PCR with primer specific to sugarcane EST-SSR (E-30-1 and E-31-1). The presence of these sequences has revealed the potential as bioenergy crop. Similarly, genetic engineering experiments were conducted to develop transgenic sweetpotato expressing synthetic lytic antiviral peptides, which are capable of inhibiting HIV progression to address epidemic diseases issues such as AIDS responsible for millions of deaths annually. Seven transgenic plants have been confirmed through molecular tests, are currently subjected to challenge for efficacy against viral replication. The effects of sweetpotato green diets on apolipoproteins were also investigated to take advantages of the full potential of the vegetable crop. What opportunities for training and professional development has the project provided? Training opportunities in terms of summer workshops, summer internships, and student’s workstudies were provided respectively, some 200 K-12 educators and farmers, 150 K-12 students, and several undergraduate and graduate students. About 10 student theses have obtained under the project. Several visiting scientists and scholars were trained under the research realm of this project. How have the results been disseminated to communities of interest? The results obtained from this project were not only published in peer-reviewed journals, but also disseminated through several presentations given at scientific and professional agriculture meetings, such as Plant and Animal Genome conference, the sweetpotato collaborator group meeting, ARD, the Society for Invitro Biology, PAWC and ABA meeting in England. Many on-farm demonstrations were conducted to share the outcomes of the vegetable production components. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
For objective 1, phylogenetic relationship of species of genus Arachis based on genic sequences displayed four clades of Arachis species. Species A. triseminata was genetically isolated from all other Arachis species studied, and formed the basal clade with A. retusa and A. dardanoi from the most ancient sections Extranervosae and Heterminatae, respectively. Species of section Arachis formed a separated single clade from all other species, within which species having B and D genome clustered in one subgroup while three species characterized with an A genome grouped together in another subgroup. A divergent clade including species from five sections was sister to the clade of section Arachis. Between the sister clades and the basal clade, there was a clade containing species from the more advanced sections. Molecular phylogeny was generally congruent with the convention-based classification except the divergent lineage in this study. For objectives 2 and 3, varietal screening and isolation of resistance gene homologs (RGHs) provided sufficient transcript derived fragments, and insight into the molecular mechanism underlying the storage root formation and development. Thus, high throughput transcriptome sequencing is needed to generate enormous transcript sequences from sweetpotato root for gene discovery and molecular marker development. The use of disease resistant cultivars is beneficial for increasing overall crop yields; thus, a molecular approach was used to compare the expression levels of the pathogenesis-related protein (PR) and PSII. The resistant variety (Jewel) demonstrated a higher expression of PR and PSII compared to the susceptible cultivar (Whatley La) indicating its superior capability in disease resistance and plant growth and development. PRP and PSII expressions were higher in the 2 sweetpotato cultivar in comparison to the susceptible cultivar (Whatley LA) but lower than that of Jewell; these results may promote efficiency in breeding to promote the sustainable production of sweetpotato. In addition, a rapid detection assay was performed on okra galls to identify the presence of the root knot nematode Meloidogyne incognita through PCR of its DNA extracted from okra galls; thus, providing a rapid molecular diagnostic test for identification of the root-knot nematode. For harvest index, PSII, a major protein involved in photosynthesis and is an indicator of overall plant growth and development along with Beta-Actin, a house keeping reference gene, were steadily expressed in all samples regardless of treatment. Based on the genetic expression analysis the fish fertilizer treatment was more beneficial for plant growth and development compare to conventional fertilizer. Fish fertilizers provide an excellent source of nutrition for plants and the soil with decrease in use of ground fertilizer with substantial cost savings.
Publications
- Type:
Other
Status:
Published
Year Published:
2013
Citation:
Marian D. Quain and Marceline Egnin. 2013. Agricultural Biotechnlogy In Ghana The Status: In Genetically Engineered Crops In Developing Countries Eds.: DVR Reddy, P Ananda Kumar, P Lava Kumar, G Loebenstein &C Kameswara Rao. Studium Press LLC, Houston, USA. pp. 377-397.
- Type:
Conference Papers and Presentations
Status:
Submitted
Year Published:
2013
Citation:
Owens, Emma, M. Egnin and B. Philip. 2013. Metabolic Profiling Of Apolipoproteins A4, B100 And E In Obese And Non- Obese Individuals From Select Black Belt Counties Of Alabama. 69th PAWC Proceeding, pg 79-90.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2014
Citation:
Guohao He, Noelle A. Barkley, Yongli Zhao, Mei Yuan, C.S. Prakash. (2014). Phylogeny of The Genus Arachis Based On Genic Sequences. Genome (accepted).
- Type:
Conference Papers and Presentations
Status:
Submitted
Year Published:
2013
Citation:
Samuels, S., M. Egnin, J. Jaynes. 2013. Development of Plant-based Therapeutic Treatment Regimen against HIV Replication. 69th PAWC Proceeding, pg 112-120.
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Baozhu Guo, Manish K. Pandey, Guohao He, Xinyou Zhang, Boshou Liao, Albert Culbreath, Rajeev K. Varshney, Victor Nwosu, Richard F. Wilson, H. Thomas Stalker. (2013). Recent Advances in Molecular Genetic Linkage Maps of Cultivated Peanut (Arachis hypogaea L.). Peanut Science 40: 95 106.
- Type:
Journal Articles
Status:
Other
Year Published:
2013
Citation:
Hui Wang, Manish K. Pandey, Lixian Qiao, Hongde Qin, Albert K. Culbreath, Guohao He, Rajeev K. Varshney, Douglas R. Cook, and Baozhu Guo. (2013). Genetic Mapping and QTL Analysis for Disease Resistance Using F2 and F5 Generation-based Genetic Maps Derived from Tifrunner � GT-C20 in Peanut (Arachis hypogaea L.). Plant Genome 6:3, doi:10.3835/plantgenome2013.05.0018.
- Type:
Other
Status:
Submitted
Year Published:
2013
Citation:
Melissa Johnson, Ralphenia D. Pace, Wendell H. McElhenney, Marceline Egnin, Temesgen Samuel and Norma L. Dawkins. 2013. Evaluation of a Dietary 25:1 Omega-6/Omega-3 Fatty Acid Ratio in SHRs Fed Traditional and Nontraditional Vegetables: Part I. Nutrients 2013, 5, 1-x manuscripts; doi:10.3390/nu40x000x. ISSN 2072-6643.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2014
Citation:
L. K. Milad, Angel Grady, OSAGIE IDEHEN2, Steven Samuels, Gregory Benard, Marceline Egnin, Desmond Mortley, Conrad Bonsi, Crystal Lee, Inocent Ritte. Comparative Analysis of PSII Transcript Expression as Harvest Index Indicator in Crop Under Different N Source. SIVB Symposium 2014, Savannah, GA
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2014
Citation:
Parks, Lashachiah Desmond G. Mortley, Marceline Egnin and Moabin Tu. Influence of Added Nutrients in the Fermentation process of Sugar Cane and Sweet Sorghum Varieties for Ethanol Concentration. SIVB Symposium 2014, Savannah, GA
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2014
Citation:
Steven Samuels, Marceline Egnin, Nashar Toufic, Jesse Jaynes, C S Prakash and Inocent Ritte. Ingineering Sweetpotato Expressing Lytic Peptides for the Potential Inhibition of HIV Replication. SIVB Symposium 2014, Savannah, GA. Winner 3rd Place. SIVB Symposium 2014, Savannah, GA.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2014
Citation:
Gregory C. Bernard, Marceline Egnin, Steven Samuels, Desmond Mortley, William Witola, Kathy Lawrence and Conrad Bonsi. Molecular Fingerprinting of Transcripts involved in Host Response to disease in Developing Sweetpotato Storage Roots. SIVB Symposium 2014, Savannah, GA.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2014
Citation:
Steven Samuels, Marceline Egnin, Nashar Toufic and Jesse Jaynes. Engineering Sweetpotato Expressing Lytic Peptides for the Potential Inhibition of HIV Replication. George Washington Carver Legacy Symposium. April 23, 2014, Iowa State University.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2014
Citation:
Gregory C. Bernard, Marceline Egnin, Steven Samuels, Desmond Mortley, William Witola, Kathy Lawreence and Conrad Bonsi. Molecular Fingerprinting of Transcripts involved in Host Response to disease in Developing Sweetpotato Storage Roots. George Washington Carver Legacy Symposium. April 23, 2014, Iowa State University
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2014
Citation:
Basak, S., C Prakash, G He, M Egnin & E Sacks. 2014. Genetic Diversity Among Bioenergy Grass Miscanthus In The Natural Populations of US Using Molecular Markers. George Washington Carver Legacy Symposium. April 23, 2014, Iowa State University.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2013
Citation:
Steven Samuels, Marceline Egnin, Jesse Jaynes and Grgory Bernard. Development of A Plant-Based Treatment Regimen against HIV Replication. CREATE-IGERT Symposium January 2013, UC-Davis, CA.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2013
Citation:
Gregory C. Bernard, Marceline Egnin, Steven Samuels, Desmond Mortley, William Witola, and Conrad Bonsi Endpoint cDNA expression analysis of molecular markers involved in host defense to disease in developing sweetpotato storage roots. CREATE-IGERT Symposium January 2013, UC-Davis, CA
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2013
Citation:
G. C. Bernard, M. Egnin, C. Bonsi, W. Witola, and V. Khan. Differential gene expression in developing sweetpotato storage roots in response to infection by the root knot nematode Meloidogyne spp. ARD Symposium 2013, Jacksonville, FL.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2013
Citation:
SB Samuels; M Egnin; JM Jaynes. Development of a Plant-Based Treatment Regimen against HIV Replication in Sweetpotato [Ipomoea Batatas (L.) Lam] Expressing Synthetic Lytic Peptide Genes Jc41n and Jc41n. ARD Symposium 2013, Jacksonville, FL.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2013
Citation:
SB Samuels; M Egnin; JM Jaynes; SM Traore. Development of A Plant-Based Treatment Regimen against HIV Replication. RCMI 2013 Symposium in Puerto Rico
|
Progress 10/01/11 to 09/30/12
Outputs
Target Audience: Target audience includes global biotechnology, genomic and breeding communities involved in peanut, sweetpotato, root and tuber, bioenergy, legume and vegetable research and production. The realm of this research serves as a platform for scientists, scholars, growers, farmer and extension specialists to benefit from the outcome and similarly impact student learning and educators around Tuskegee University community. The ongoing sweetpotato nematode work will benefit local farmer in their production practices. Changes/Problems: Biofuel production has become an important alternative to the use of existing natural resources for energy. Bamboo contains high cellulose content, good disease resistance and can be propagated in wide array of climates, which are useful characteristics for biofuel conversion. To identify plants best suited for bioenergy production, Moso Bamboo along with Miscanthus, Sugar Cane, Gamma Gras and 2 Sweetpotato (TUO2, DMO1-158-204) cultivars Genomic DNA were subjected to PCR with primer specific to sugarcane EST-SSR (E-30-1 and E-31-1) and PSII. PCR amplification of the Photosystem II gene fragment serves as a reference and was present in all samples. A 400 bp amplicon was generated in all samples with E-30-1, while three distinct amplicons (250bp, 300bp and 400bp) were obtained with E-31-1. These varying size PCR product sequences may be due to nucleotide mutation within the SSR motifs and flaking sequences with E-31-1. Earlier work confirmed the nature of these EST-SSR sequences as closely related to energy specific cellulose synthase gene for cellulosic characteristic. The presence of these sequences shows the potential as bioenergy crop. What opportunities for training and professional development has the project provided? Our research, especially under objectives 4 and 5 has provided training opportunities in hands-on field and laboratory work for several PhD, graduate and undergraduate students as well as visiting scientists, in all aspect of the projects though. Additional summer workshops for K-12 students and educators were provided for in biotechnology exploration. This year, 5 high school students participated in our summer program for 8 weeks, working on some of the objectives accomplished so far. The scientific and farmer communities at large have benefited from several presentations given at scientific and professional agriculture meetings, such as Farmers Conference, Plant and Animal Genome conference and PAWC. Ongoing collaborative research with UC-Davis provided professional development practicum to students. How have the results been disseminated to communities of interest? The various results were disseminated to community of interest through publications, and presentations: What do you plan to do during the next reporting period to accomplish the goals? Several aspects of the ongoing project dealing with the biosafety assessment of the transgenic plants, the disease resistance molecular screening via cDNA-AFLP of newly developed cultivars will be performed. The next generation project centers around bioenergy; thus HighThroughput functional genomic work will be done on bamboo Miscanthus and sugarcane as well sweetpotato.
Impacts
What was accomplished under these goals?
Ojective 1: The better understanding of the phylogenetic relationships between Arachis species, accessions were accomplished through sampling from all nine taxonomic sections and seven genic sequences to reconstruct the phylogeny of the genus Arachis. Objectives 2 and 3: Test for pathogenesis related proteins as reference for disease management were performed with sweetpotato storage root development and Okra subjected to disease challenge for 56 days to provide an insight into disease response to nematode infection. Several cultivars were evaluated for two populations of nematode infections and resistance screening of infection and reproduction were assessed. Additionally a molecular diagnostic assay was performed to identify and detect the presence of Meloidogyne incognita in okra DNA in the galls of infected plants. Enhance understanding of harvest index in sweetpotato, tomato, okra and bell pepper were also conducted under culture procedures with different fertilizers applied during growth and formation of storage root. Molecular screening of total cDNA generated substantial fraction of transcript sequences, which can be used to target disease resistance response and profile novel genes during storage root formation and development and make it possible to further characterize gene expression profiles during these processes. Objectives 4 and 5: Molecular screening of transgenic plants developed under the project continue as well outreach activities geared to plant breeding, biotechnology and related aspects.
Publications
|
Progress 10/01/10 to 09/30/11
Outputs
OUTPUTS: Objective: Peanut genome structure. Construction and improvement of a genetic map for peanut (Arachis hypogaea L.) continues to be an important task in order to facilitate quantitative trait locus (QTL) analysis and the development of tools for marker-assisted breeding. A total of 324 markers were anchored on the integrated map covering 1,352.1 cM with 21 linkage groups (LGs). Combining information from duplicated loci between LGs and comparing with published diploid maps, seven homologous groups were defined and 17 LGs (A1-A10, B1-B4, B7, B8, and B9) were aligned to corresponding A-subgenome or B-subgenome of diploid progenitors. Objective: Cloning, characterization and utilization of gene associated with diseases. RGA based degenerate primers have been designed to screen and characterization disease resistance motifs in field grown sweetpotato cultivar exhibiting multiple nematode, insect and virus resistance. Objective: Use of Comparative genomic to characterize agronomic gene expressions. We sought to predict the yield of Bell pepper through system analyses of metabolic interaction in nutrient input and photosynthetic activities utilizing photosystem II (PSII) transcript as harvest index. Accumulation of PSII mRNA in developing Bell pepper plants subjected to: no fertilizer (P1), broiler liter (P2), conventional fertilizer (P4) and fish compost (P14) were profiled through Real-Time quantitative analysis with SYBR green. Total RNA was isolated from all 4 treatment samples, reversed transcribed (RT-PCR) into complementary DNA (cDNA) then subjected to Real-Time PCR (qPCR) with PSII primers along with positive control and no template and enzyme controls (NTC). Objective: Develop transgenic screen methods to engineer new cultivar. Efficient somatic embryogenesis and genetic transformation systems, applicable across a range of sweetpotato genotypes to facilitate the genetic manipulation and improvement has led to 17 PCR positive independent transgenic plantlets with waxi and jc41N and jc41ND genes. Integration of the transgenes is currently under investigation through Southern and qRT-PCR analyses. Objective: Outreach activities. Training activities were undertaken with laboratory essential techniques and bioinformatics to improve and leverage student involvement in all research and outreach component of this project. Additionally, public lectures, invited seminars, meetings with policy makers and other stake holders, Internet websites including social networks were utilized as avenues for disseminating our activities to enhance the societal awareness of food biotechnology issues around the world. Collaborative research procedures were performed for yam and sweetpotato germplasm conservation and improvement with Ghana. PARTICIPANTS: Faculty Members: M Egnin, G He, D Mortley, C S Prakash, C Bonsi, J Jaynes, J Jackson Research Associates: K Nyyiawung, L Odom Graduate Students: Steven Samuels, Chris Bernard Collaborators: Ming Gao (Alcorn), Marian Quain (Ghana), Douglas Cook and R. V. Penmetsa in UC Davis, R. K. Varshney at ICRISAT, India, D. J. Bertioli at the University of Brasilia, Brazil, and S. Isobe at Kazusa DNA Research Institute, Chiba, Japan have been involved in genetic mapping of peanut. One research assistant, one visiting scholar, and two undergraduate students in Tuskegee University were trained in the project. TARGET AUDIENCES: The outcome of this project is targeted to the biotechnology, genomic and breeding communities especially in Peanut, sweetpotato, root and tuber, legume and vegetable research and production. Our project is used as a platform to impact student learning, educator and farmers around Tuskegee University community - local, regional, state and international. Target audiences benefited from two of the developed peanut genetic maps published. The scientific and farmer communities at large benefited from several presentations given at scientific and professional agriculture meetings, such as Plant and Animal Genome conference, the sweetpotato collaborator group meeting, ARD, PAWC and ABA meeting in England. The root and tuber crop communities may benefit from our yam procedures for germplasm conservation and improvement of other species, using limited laboratory resources. PROJECT MODIFICATIONS: Sweetpotato endogenous enzyme impact on ethanol production was investigated using twenty gram of fresh root samples from high starch cultivars TIB4 (29.2% starch yield) and DMOI 158 097 (28.4% starch yield) with unit amylase concentrations of 20.6 and 15.88 mU per ml respectively. The frozen samples were blended with 40 ml of water and 0.050 ml of 1M NaOH then the temperature was quickly raised to 68 degree C to gelatinize starch by native alpha and beta amylases. After gelatinization, samples were hydrolyzed at different temperatures (60, 65 and 70 degree C) for 2 hours at 200 rpm, quickly cooled, and pH adjusted to 4.5 with nitric acid, and 400mg yeast added for anaerobic fermentation at 30 degree. Ethanol concentration after HPLC was estimated from different ethanol standard concentrations. Sample hydrolyzed at 65 degree C had the highest ethanol concentration, followed closely by the 70 degree C treatment. The 60 degree C samples had the lowest concentrations probable due to the synergistic work of alpha and beta amylases between 60 and 70 degree C. Based on starch and amylase content, TIB4 had higher ethanol concentration compared to DMOI 158 097 at 60 degree C but not at 70 degree C. Samples hydrolyzed immediately after blending had lower concentrations compared to those stored before hydrolysis. These results show the potential of high endogenous enzyme levels in reducing cost of bioethanol production from sweetpotato through biotechnology. Successful germplasm conservation, organogenesis and transformation protocol have been optimized and adapted for engineering yam quality and productivity. Yam, Dioscorea rotundata, is one important staples and sources of carbohydrate in the diet in Sub-Saharan African sub-region. The crop has several problems: post harvest, availability of the edible planting materials and high cost of labor for cropping; therefore, needs to be improved using modern biotechnology. We have developed simple, cost-effective and viable cryopreservation methods for Dioscorea rotundata. The methodology requires a simple vitrification protocol that incorporate pregrowth of the donor plant on sucrose-supplemented medium for five weeks, preculture on 0.3 M sucrose supplemented medium for 5 days, PVS2 solution for 40 min, rapid cooling in liquid nitrogen or slow cooling to -80 degree C. We have also induced shoot regeneration in yam with leaf petiole cultured on MS medium supplemented with 0.2 mg 2,4-D for 3 days, and transferred to MS medium containing TDZ alone or in combination with 2iP. Shoot regeneration was observed within 21 days on all media; however, the highest shoot regeneration, 7-9 shoots developing per explant were obtained on media supplemented with TDZ and 2iP. Very high efficiency of transformation (25-65 percent) was obtained when petiole explants were subjected to Agrobacterium-mediated transformation using strain C58 and EHA101 harboring a binary plasmid containing gus-A intron gene under the transcriptional control of CaMV35S promoter.
Impacts
The peanut genome work established a comparative integrated map from two cultivated x cultivated recombinant inbred line (RIL) mapping populations for transferability and mapping tomato spotted wilt virus (TSWV) resistance trait in peanut. One reciprocal translocation was confirmed in the tetraploid-cultivated peanut genome. Several chromosomal rearrangements were observed by comparing with published cultivated peanut maps. High consistence with cultivated peanut maps derived from different populations may support this integrated map as a reliable reference map for peanut whole genome sequencing assembling. Further, two major QTLs for TSWV resistance were identified for each RILs, which illustrated the application of this map. Ahead of the RGA work, we have successfully isolated Total RNA from leaves and generation cDNA using a bifunctional NotI-oligodT primer. These will be subjected to gene profiling for RGA motifs. Gene expression profiling to elucidate molecular system analysis exhibited successful amplifications of PSII cDNA in all treatments, and more importantly showed the highest transcript levels in fish fertilizer treatment of Bell pepper correlating to canopy and source levels in the field. The fluorescence trace of the fish fertilizer bell pepper plant with PSII primers has a CT value of 15.905 the closest to the control PSII DNA (9.6), and reached the threshold line before the other fertilizer-treated plants indicating more total PSII mRNA accumulated in the P12 plant to generate such high cDNA levels than the others. The P4 (NPK) with a CT value of 16.45 closely followed as the second highest level, while P2 had the third highest level with the least in P1. Thus, Fish fertilizer had more of an impact on the productivity of the Bell pepper based on the Threshold value closest o the control PSII DNA, an indicator that fish fertilizer yielded the best harvest index. Beauregard, NCC58 and PI 318846-3 (D-3) were more amenable to Agrobacterium transformation (68- 87%) than other cultivars tested. Disarmed Agrobacterium tumefaciens strains EHA101 and EHA105 were superior in facilitating the transfer of transgenes to sweetpotato cells than strains and C58. Transgenic plantlets have been screened for the presence waxi and jc genes. Further analyses to test the efficacy of the peptide on HIV in infected cells lines are currently in the planning phase, while waxi sweetpotato will be screened for starch and amylase levels. We have strived to foster better understanding of biotechnology both in the US and outside the country as well as impact our underserved minority students academic development. Our efforts have also contributed to the global policy agenda in agricultural biotechnology; advanced dialog and discussion especially among scientists and other stakeholders on agricultural biotechnology, and in creating an awareness of benefits and safety of this technology through science-based evidence.
Publications
- Qin, H., S. Feng, C. Chen, Y. Guo, S. Knapp, A. Culbreath, G. H. He, M. Wang, X. Zhang, C. C. Holbrook, P. Ozias-Akins, B. Guo. 2011. An integrated genetic linkage map of cultivated peanut (Arachis hypogaea L.) constructed from two RIL populations. Theor Appl. Genet. DOI 10.1007/s00122-011-1737.
- Quain, M. D., M. Egnin, B. Bey, R. Thompson and C. Bonsi. 2011. Transgenic potential of Dioscorea rotundata using Agrobacterium-mediated genetic transformation, GM Crops: From basic research to application. Aspects of Applied Biology 110:71-79.
- Quain, D. M., P. Berjak, E. Acheampong and M. Egnin. 2011. Cryopreserving Vegetatively Propagated Tropical Crops, the Case of Dioscorea Species and Solenostemon rotundifolius. Cryopreservation Book 1, ISBN 979-953-307-300-1.
- Samuels, S., M. Egnin, J. Scoffield, B. Bey, S.Traore, C. S. Prakash, J. Jaynes and J, Jackson. 2011. Somatic Embryogenesis and Genetic Transformation Of Multiple Sweetpotato [Ipomoea batatas L. (Lam)] Cultivars for Enhanced Productivity, Nutritional and Health Values. Proceeding of the National Sweetpotato Collaborators Group Progress Report.
- Prakash, C. S. 2011. GM in the Media. GM Crops 2: 1-2.
- Wang, Hui, R. V. Penmetsa, M. Yuan, L. Gong, Y. Zhao, B. Guo, A. D. Farmer, B. D. Rosen, J. Gao, S. Isobe, D. J. Bertioli, R. K. Varshney, D. R. Cook, G. H. He. 2011. Development and characterization of BAC-end sequence derived SSRs, and their incorporation into a new higher density genetic map for cultivated peanut (Arachis hypogaea L.). BMC Plant Biology (accepted).
- Nyiawung, K. Z., D. Mortley, M. Egnin, C. Bonsi, and B. Vaughan. 2011. Sweetpotato In: Handbook of Bioenergy Crop Plants K10867-C031.indd, 31:734-742.
- Prakash, C. S. 2011. Agricultural Biotechnology. In FOOD: IN CONTEXT. (Textbook Chapter) Published by Gale, a part of Cengage Learning.
- Odom, L., C. Bonsi, R. Ankumah, J. Jaynes, M. Egnin, L. Ogden, and D. Mortley. 2010. Effect Of Antimicrobial Synthetic Peptide D4e1 On Infestation Of Cotton Seedling Disease And On Soil Microbial Diversity, In Proceeding of the 66th Annual Professional Agriculture Workers Conference-PAWC - Facing Global Crisis: Local Solutions To Energy, Food and Persistent Poverty: Edited by Nii O. Tackie, Tasha M.Hargove, Robert Zabawa, and Walter A. Hill. pg. 226-235
- Varshney, R., C. Gowda, T. Radhakrishnan, G. Bhimana, M. Pandey, V. Sujay, R.Koppolu, V. Vadez, S. Nigam, H. Upadhyaya, Y. Khedikar, S. Isobe, G.H. He, D. Bertioli, S. Knapp, and D.R. Cook. (2011). Towards integrating genomics in breeding in groundnut (Arachis hypogaea L.). January 15-19, 2011, Presentation abstract at Plant & Animal Genome XIX, San Diego, CA.
- Penmetsa, Varma, B. Rosen, N. C. Garcia, J. Gao, B. K. Sarma, S. Datta, S. L. Vail, L. Garzon, B. Vandenberg, J. Woodward, M. Blair, M. Nelson, D. Bertioli, G. D. May, R. K. Varshney, M. Yuan, G.H. He, G. E. Bruening, and D.R. Cook. (2011). Comparative genetic maps. January 15-19, 2011, Presentation abstract at Plant & Animal Genome XIX, San Diego, CA.
- Qin, Hongde, Suping Feng, Charles Chen, Steven Knapp, Albert Culbreath, Guohao He, Mingli Wang, Xingyu Zhang, Corley Holbrook, and Baozhu Guo. (2011). An integrated linkage map for cultivated peanut (Arachis hypogaea L.) derived from two RILs populations. January 15-19, 2011, Presentation abstract at Plant & Animal Genome XIX, San Diego, CA.
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Progress 10/01/09 to 09/30/10
Outputs
OUTPUTS: Objective: Cloning, characterization and utilization of genes associated with sweetpotato storage root development and regeneration. We investigated the relationship between t-zeatin riboside and nitrogen (N) nutrition during storage root initiation. Stem cuttings were grown hydroponically in nutrient solution containing either 1.5, 3.0, or 4.5 mM N and harvested at 7, 10, 14, 18, 21 and 28 days. Generally, fresh weight, was greater among plants receiving the higher N level regardless of variety used. Root diameter increased steadily for both cultivars regardless of N levels. Root weight and diameter were scored and samples were collected for t-zeatin riboside analysis and gene expression profiling. The work aimed to identify genes expressed during sweetpotato storage root secondary growth was analyzed through Real-Time PCR (qPCR) of developing sweetpotato root cDNA with six genes specific primers (Sporamin, Gigantea, tumor related protein, J8, expansin and invertase). Further studies will be done to better understand this relationship. Objective: Peanut genome structure. A large set of SSRs (1450) was developed from BAC end sequences of cultivated and wild-diploid species of Arachis that includes 142 SSRs in resistance gene homolog (RGH) containing BACs (collaborated with UC Davis). Together, this new set and other public available SSRs represent about 6,000 SSRs that were screened for genetic polymorphism. Screening result has showed 408 polymorphic SSR loci between two parental genotypes Tifrunner and GT-C20 of a mapping population developed by USDA-ARS at Tifton, GA. Among these polymorphic loci, 347 were distributed into 24 linkage groups, and cover 1,789.3 cM with an average distance of 5.1 cM between adjacent markers. Three RGH-related SSR loci were anchored in three linkage groups. Objective: Outreach activities. The focus was on training student and enhancing the societal awareness of food biotechnology issues around the world with public lectures, invited seminars, meetings with policy makers and other stake holders. Outreach through internet websites and social networks was an important avenue for disseminating information and promoting discussion on this subject among stakeholders such as scientists, policy makers, activists and journalists. PARTICIPANTS: Faculty Members: Marceline Egnin, Guohao He, Desmond Mortley, Conrad Bonsi and C. S. Prakash Research Associate: Karine Nyiawung Graduate Students: Steven Samuel and Lakisha Odom Collaborators: Douglas Cook, Benjamin Rosen, Ming Gao and Carol Harrison This Project along with our general outreach program provided support for Biotech and biosafety training to students and African scientists. The mapping population was provided by the collaborator Dr. Baozhu Guo (USDA-ARS at Tifton, GA). Development of SSRs from BAC end sequences was obtained in collaboration with Dr. Doug Cook of UC Davis. TARGET AUDIENCES: The outcome of this research is targeted to the biotechnology and genomics research communities especially National Sweetpotato Collaborators Group, Sweetpotato Growers Association, Peanut Research Community, Legume Genomics community, Root and Tuber Crop researchers, small farmers, K-16 educators and students in underserved communities. The project was also used as a forum to train several undergraduate and graduate students in molecular biology and genomics related to the project, specifically in PCR technique, qRT-PCR, ELISA, gel electrophoresis, data analysis, and mapping methods. We have attended and gave presentations at peanut annual meeting and SIVB. One graduate student and one visiting scientist from China worked on genetic mapping. The peanut genetic linkage map was made available to peanut research community. PROJECT MODIFICATIONS: Genetic engineering experiments were conducted to develop transgenic sweetpotato expressing synthetic lytic antiviral peptides JC41N and JC41ND, which are capable of inhibiting HIV progression to address epidemic diseases issues such as AIDS caused by human immunodeficiency (HIV) virus are responsible for millions of deaths annually Two de novo synthetic gene constructs were designed and synthesized to facilitate cloning in bacteria and accumulation in plants without lethality. Fifty-five kanamycin resistant embryos of D-3 transformed with jc41N and jc41ND genes were obtained and germinated on MM supplemented with Timetin 100mg/l and Kanamycin 12.5mg/l, resulting in twenty four (24) putative transgenic plantlets.
Impacts
Gene expression profiling and characterization along with molecular markers and genetic linkage maps are the prerequisites to the elucidation of molecular processes in function assignment, and to marker-assisted selection in plant breeding, respectively. Preliminary hydroponic studies impacted the maximum diameter of 2 mm for J6/66 obtained at day 21 among plants receiving 1.5 mM N (N1) followed by plants receiving 3mM N (N2). Maximum root diameter for NCC-58 exceeded 2 mm on day 21 and 3 mm on day 28 among plants treated with 3 mM N. As root diameter increases, the concentration of t-zeatin riboside in J6/66 increased at all levels of N and declined at day 28 except among plants receiving 1.5 mM N (N1). A similar trend was observed for NCC-58 except among plants receiving 3 mM N (N2). These preliminary results suggest that there may be some impacts of N levels in suppressing the concentration of t-zeatin riboside at both the 1.5 and 4.5 mM N especially for NCC-58. Additionally, the identification of storage root-specific expressed genes related to parenchyma cells development (starch, protein and carotenoid accumulations) from the quantitative gene expression analyses supported the molecular and physiological processes occurring during sweetpotato storage root formation. Nine cDNA-AFLP clones of the developing storage roots were catalogued upon BLAST by molecular and cellular processes such as cell gene regulation, division and expansion, signal transduction, tumorigenesis, sporamin accumulation. The outcomes of the peanut genomics work to date, is the development of the highest density genetic linkage map in a single mapping population of cultivated peanut (manuscript in preparation). Therefore, the SSR markers developed and genetic linkage map constructed here have provided useful genetic and genomic tools for peanut research. The database of a large number of SSRs mined from BAC end sequences was submitted to GenBank. This genetic linkage map was presented and shared with peanut research community at the conference of America Peanut Research and Education Society.
Publications
- Wang, Y., B. Rosen, J. Scoffield, M. Egnin, D. Mortley, S. Steiner, D. R. Cook and G. He. 2010. Isolation and analysis of resistance gene homologs in sweetpotato. Plant Breeding 129: 519-525.
- Yuan, M.; Gong, L.; Meng, R.; LI, S.; Dang, P.; Guo, B. and He, G. (2010). Development of trinucleotide (GGC)n SSR markers in peanut (Arachis hypogaea L.). Electronic Journal of Biotechnology, vol. 13, no. 6. http://dx.doi.org/10.2225/vol13-issue6-fulltext-6. Peggy, V., J. Xu, M. Goore, J. Smolnik, M. Egnin, T. Jones, L. Haile. 2010. The Relationship between Diabetes, Obesity and Iris Markings in African Americans in Montgomery County, Alabama. Submitted to the Journal of Irridology
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Progress 10/01/08 to 09/30/09
Outputs
OUTPUTS: A total of 54,000 BAC clone-ends was sequenced in peanut, from these BAC end sequences (BES) 1152 SSRs were identified (GenBank accession number FI499377-FI503061). Among 1152 SSRs, 142 were found related to RGH-containing BAC clones. These RGH related SSRs will be placed onto linkage map. About 225 distinct sweetpotato NBS sequences with similarity to known RGH genes were identified. Additional 50 sweetpotato RGHs were mined from the public genomic sequence database. Thus, a total of 275 RGH sequences were obtained using both PCR-based method and data-mining approach, from which 237 were non-TIR sequences organized into 35 singletons and 35 groups after reduced to 90 percent nucleotide identity, and 38 were TIR sequences divided into three primary phylogenetic clades. Physical map of RGAs in peanut identified 589 unique RGHs likely to represent the preponderant majority of resistance genes in cultivated peanut. Using the BAC library, a total of 3,784 BAC clones containing RGH were identified by hybridization with RGH probes. Cultivar NCC-58 was grown in nutrient film (NFT) and culture media to investigate changes in gene expression during sweetpotato storage root development. RT-PCR was performed to generate cDNA for AFLP identification of transcripts differentially (TDF) expressed. EcoRI and MseI primer combinations resulted in 48 selected TDFs from NFT and MSR. Both exhibited the same profile in four different expression patterns: up-regulated, decreased expression, transient expression and constitutive expression. The time point for peak expressions was delayed with invitro-grown when compared to NFT. Based on the BLASTX homology analyses, sweetpotato TDF predicted function were homologous at 77 and 78 percent, respectively to: Ipomoea batatas mRNA clone IT443-3-end sequence of Kokei-14 mature tuberous root, and storage root cDNA library. Primers specific to sporamin precursor, GIGANTEA and Calmodulin-like protein of Arabidopsis thaliana, Lycopersicon esculentum Expansin precursor and Hordeum vulgare Translationally controlled tumor protein identified quantitative and differential expression patterns in storage roots versus fibrous and fibrous to storage root. The impact on insect vectors of TSWV and incidence of disease symptoms and subsequent fruit yield were screened in tomatoes and peppers grown in experiments mulched with Red, Violet, Aluminum coated, or White-on Black agricultural plastics. Fruit yield of pepper was highest under white-on black, followed by aluminum-coated mulches and lowest for violet mulches. A similar response was observed for tomatoes but with identical yields between white-on black and aluminum coated mulches. The most common insect group was Diptera. Fewer insects were captured on yellow sticky traps in aluminum-coated plats followed by white-on black-mulched plots. Symptoms of TSWV were seen on pepper fruits in some plots mulched with violte plastic. A two days field visit afforded the students the opportunity to experience real world applications for many of the techniques utilized in our research and the steps involved with evaluating transgenic crops for commercial use. PARTICIPANTS: Participants Faculty Members: Marceline Egnin, Guohao He, Desmond Mortley, BJ Min, Jesse Jaynes, C. S. Prakash and Jacquelyn Jackson Research Associate: Frieda Sanders Graduate Students: Sy Traore, Tarek Radwan, Lakisha Odom and Steven Samuel Collaborators: Douglas Cook, Benjamin Rosen, Ming Gao and Carol Harrison Monsanto corporation This Project along with our general outreach program conducted biosafety workshops, student training and field visits. The workshops focused on biotech and Biosafety activities previously developed with an infusion of Biofuels and BioMedical activities. TARGET AUDIENCES: The outcome of this research is targeted to the biotechnology and genomics research communitie, especially Peanut Research Community, Legume Genomics community, sweetpotato growers association, root and tuber crop researchers, small farmers, K-16 educators and students in underserved communities. PROJECT MODIFICATIONS: With the demand for biomass based ethanol production such as starch, and the known fact that fermentable sweetpotato carbohydrate yield is grater than that of field corn we have modified objective 2 to include the screening of sweetpotato cultivars for starch and alpha amylase contents. Preliminary results showed that three sweetpotato cultivars (C, G, and M) had respectively, 28.4, 29.2 and 28.8 percent levels of starch with high potential for starch-based bio-ethanol. In this study, starch physical properties and enzyme activity of 14 sweetpotato cultivars were investigated. The color range of freeze-dried sweetpotato meal was white to yellow. White flesh showed higher L value while yellow flesh showed higher b value. In alpha amylase activities, cultivars E, G, M showed higher than 20mU/mg. Amylose levels ranged from 18.0 to 19.4, while that of amylopectin ranged from 80.6 to 82.0. However, there was no significant difference between amylose/amylopectin ratios. Our results showed that two cultivars might have potential for bio-ethanol production based on their starch yields and alpha-amylase activity.
Impacts
The broader impact is to discover and understand gene functions, and develop crop for high productivity and better human nutrition and health. These partial genomic and biotechnology results of sweetpotato, peanut and selected crops have contributed the identification of RGHs in sweetpotato and peanut, understands genome structure/organization and key physiological and biochemical functions during storage root development. Field screening of selected crops revealed the impact of colored plastic mulches on fruit yield and incidence of tomato spotted wilt virus (TSWV) in tomatoes and peppers. These transferable visual markers will aid in the screening of resistance genes homologs related to the infectivity of TSWV) in tomatoes and peppers. The mapping of BES-SSRs will simultaneously anchor RGH-containing BAC seed points throughout peanut genomes, and enhance the utility of these resources for peanut breeding. It would also facilitate cloning of disease resistance genes by map-based cloning. These new RGH sequences provided a resource of candidate genes and molecular markers for disease resistance research in sweetpotato and peanut. The exploration of RGH diversity enables resistance gene evolutionary study has facilitated the construction of a peanut physical map of RGHs resulting in 344 contigs and 334 singletons, and may aid in the isolation of new and functional alleles. Transient and up-regulated expressions were good candidates for storage root developmental markers. Storage root initiation was confirmed between 10-14 days in NFT but delayed to 24-30 days post initial invitro culture. The comparative sequence analysis and quantitative transcript profiling results coupled with the delayed in time points for peak expressions in invitro-grown when compared to NFT at different time points early on in sweetpotato storage root formation processes has greatly contributed to our understanding of sweetpotato harvest index. A site visit to The Monsanto Learning Center in Scott, Mississippi was extremely informative and educational. Students and faculty were especially interested in how transgenic screens in confined field trials were conducted and how they are used to collect data necessary for evaluating the environmental risks and crop performance. Information was provided concerning the stages of evaluation of transgenic candidate lines, the regulation and tracing of plants tested in the field (permits required at the various phases of assessment). It was enlightening for the students to realize the amount of documentation and tracking that goes into dealing with transgenic crops and how much accountability institutions undergo to ensure that there is no risk of unintended release of transgenic plant material. In addition, different measures taken for preventing dissemination of new genes from crops such as corn and soybean were discussed.
Publications
- Wang, Yu, Benjamin Rosen, Jessica Scoffield, Marceline Egnin, Desmond Mortley, S. Steiner, Douglas R. Cook, and Guohao He . 2009. Isolation and analysis of resistance gene homologs (RGHs) in sweet potato. Plant Breeding DOI: 10.1111/j.1439-0523.2009.01711.x.
- Varshney, R. K., R. V. Penmetsa, S. Dutta, P. L. Kulwal, R. K. Saxena, S. Datta, T. R. Sharma, B. Rosen, N. Carrasquilla-Garcia, A. D. Farmer, A. Dubey, K. B. Saxena, J. Gao, B. Fakrudin, M. N. Singh, B. P. Singh, K. B. Wanjari, M. Yuan, R. K. Srivastava, A. Kilian , H. D. Upadhyaya, N. Mallikarjuna, C. D. Town, G. E. Bruening, G.H. He, G. D. May, R. McCombie, S. A. Jackson, N. K. Singh, D. R. Cook. 2009. Pigeonpea genomics initiative (PGI): an international effort to improve crop productivity of pigeonpea (Cajanus cajan L.). Mol Breeding DOI 10.1007/s11032-009-9327-2
- Varshney, R.K., D.J. Bertioli, M.C. Moretzsohn, V. Vadez, L. Krishramurthy, R. Aruma, S.N. Nigam, B.J. Moss, K. Seetha, K. Ravi, G.H. He, S.J. Knapp, D.A. Hoisington. 2008. The first SSR-based genetic linkage map for cultivated groundnut (Arachis hypogaea L.). Theor Appl Genet. 118 (4): 729-39.
- Chassy, B., M. Egnin, Y. Gao, K. Glenn, G. A. Kleter, P. Nestel, M. Newell-McGloughlin, R. H. Phipps, and R. Shillito. 2008. Nutritional and Safety Assessments of Foods and Feeds Nutritionally Improved throughBiotechnology: Case Studies. In Comprehensive Reviews in Food Science and Food Safety. Food Sci. and Techn. 7 (1):81-91.
- Dodo, H. W., K. N. Konan, F. C. Chen, M. Egnin and O M. Viquez. 2008. Alleviating Peanut Allergy Using Genetic Engineering: The Silencing Of The Immunodominant Allergen Ara H 2 Leads To Its Significant Reduction And A Decrease In Peanut Allergenicity. Plant Biotechnology Chassy, B., M. Egnin, Y. Gao, K. Glenn, G. A. Kleter, P. Nestel, M. Newell-McGloughlin, R. H. Phipps, and R. Shillito. 2008. Comprehensive Reviews in Food Science and Food Safety. J. Food Sci. 72 (9).
- Presentation: He, G.H., M. Yuan, B. Rosen, R. V. Penmetsa, D. R. Cook. 2009. Towards physical mapping of resistance gene homologs (RGHs) in peanut. January 10-14, 2009, Plant & Animal Genome XVII. San Diego, CA.
- He, G.H., M. Yuan, V. Penmetsa, R.K. Varshney, D.R. Cook. 2009. A novel set of SSRs developed from BAC-end sequences. IV International Conference of the Peanut Research Community, October 19-22, 2009, Bamako, Mali.
- Yuan, M., S.L. Li, Y. Ren, H. Wang, Y.M. Shi, S.L.Yu, G.H. He. 2009. Cloning and Characterization of a Peanut MADS-box gene isolated from flower bud. APRES annual meeting, July 14-17, 2008, Raleigh, NC.
- B. J. Min, M. Egnin, C. Bonsi, D. Mortley, S. Traore, and M. Gao. 2009. Characterization of Local Sweetpotato Cultivars as Bio-fuel Crop. Society for Invitro Biology Meeting. Charleston, SC.
- Egnin, M., F. Ssanders, H. Gao, G. He, S. Traore, and D. Mortley, B.J. Min and J. Jackson. 2009. Quantitative and Comparative Gene Expression Profiling of Differentially Expressed Genes in Sweetpotato Storage Root Development. Society for Invitro Biology Meeting. Charleston, SC.
- Odom, L., C. Bonsi, R. Ankumah, J. Jaynes, M. Egnin, L. Ogden, and D. Mortley. 2009. Effect of Antimicrobial Synthetic Peptide D4E1 on Infestation of Cotton Seed Germination Rates in the Presence of Cotton Seedling Disease. Society for Invitro Biology Meeting, Charleston, SC.
- Traore S., M. Egnin, F. Sander, S. Samuel, T. Radwan, B. J. Min and J. Jackson. 2009. Expression of Synthetic Tumor Reducing Peptide Genes in Sweetpotato as Therapeutic Drugs against Cancer. Society for Invitro Biology Meeting. Charleston, SC.
- Radwan, T., M. Egnin, P. K. Biswas, D. Mortley, S. Traore, S. Samuel and A. Shaheen. 2009. Changes in the Levels of Abscisic Acid Levels during the Germination of Citrulus colocynthis. ARD Meeting in Atlanta, GA March 2009.
- He, G.H., M. Yuan, B. Rosen, R. V. Penmetsa, D. R. Cook. 2008. Characterization of resistance gene homologs (RGHs) in peanut. IV International Conference on Legume Genomics and Genetics, December 7-12, 2008, Puerto Vallarta, Mexico.
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Progress 10/01/07 to 09/30/08
Outputs
OUTPUTS: Work under objective 1, elucidate genome structure and organization in peanut and sweetpotato using simple sequence repeats (SSR), was conducted to investigate the transferability of available soybean SSR markers to peanut. Eight hundred and sixty-eight soybean SSR primer pairs were used on varieties C20 and C3424. PCR products banding patterns were analyzed using Quanity-One gel imaging software. About 22.9 percent (199) of soybean SSR primer pairs could amplify peanut DNA. These transferable markers will be screened for polymorphism between parental peanut DNAs and added to the genetic linkage map. Under objective 2, Greenhouse, in-vitro and field trials were performed to generate plant materials for cloning, characterization and utilization of genes associated with sweetpotato storage root development and resistant gene analog (RGH) in selected crops (sweetpotato and pepper). DNA extracted from leaves yielded two hundred twenty five distinct sweetpotato NBS domains with similarity to known RGH genes were identified and organized into fourteen primary clades based on their phylogenetic analysis. An additional fifty sweetpotato RGHs were mined from the GenBank genomic sequence database. Comparative molecular and physiological analyses between invitro and hydroponic-grown sweetpotato were performed to assess changes in t-Zeatin riboside (ZR) and gene expression levels using cDNA-AFLP. Thickening potential of storage roots were studied over a 42-day and 90-day periods (4-7 sequential harvests at 4-7-day intervals). Variable ZR levels from 5-70 pmol/ml were obtained amongst all cultivars tested in the two techniques. ZR accumulation in invitro and nutrient film technique (NFT) samples followed similar accumulation patterns in storage roots of D-3 (low), TU-82-155 and J666 ZR (consistently higher) and NCC-58 (slight fluctuations). ZR levels in fibrous roots were lower than the storage root. Four selective primer combinations were tested resulting in 60 Transcript Derived Fragments (TDF) between NFT and invitro samples exhibiting a similar profile in four different expression patterns, including up-regulated, decreased expression, transient expression and constitutive expression detected in all cultivars. These findings have been presented at national meetings in Indianapolis and San Diego. PARTICIPANTS: Faculty Members: Marceline Egnin, Guohao He, Desmond Mortley and C. S. Prakash; Research Associate: BJ Min; Graduate Students: Sy Traore, Tarek Radwan, and Steven Samuel; Collaborators: Ms Carol Harrison, Ms. Mary Arrington, Dr. Edith Powell and The University of Wisconsin at Madison Genomics team. This Project along with our general outreach program conducted teacher workshops, student training and field visits. Teacher workshops focused on curriculum and activities previously developed with an infusion of highly successful Arabidopsis microarray gene chips, Biofuels and BioMedical activities. A one-week summer Youth Biotech & Genomics training for 15 rising 9-12 graders from 7 school systems in Alabama and Georgia was held. TARGET AUDIENCES: The outcome of this research is targeted to the biotechnology and genomics research community, small farmers, K-16 educators and students in underserved communities. Newly participating K-6 teachers number increased by 80%, the number of secondary and college-level teachers remained the same with a total of 45 educators from 12 Alabama counties, Georgia and Wisconsin PROJECT MODIFICATIONS: With the demand for renewable resources such as starch, a multi-billion dollar business in the US, we have modified objective 2 to include the screening of sweetpotato cultivars for starch content with great potential for bio-ethanol production. A total of 14 sweetpotato cultivars grown in the field were screened for their starch yield and physical properties. For the starch content, freshly harvested sweetpotatos were washed, peeled, chopped, and stored at negative 70 degree Celsius. Two hundred grams of frozen samples were blended with water and filtered through a 250 micron and 100 micron stainless metal sieve, and decanted. The supernatant was further decanted three times then the starch pellets were collected. The starches were dried in a vacuum chamber at 20 degree Celsius then the yield was calculated on a wet basis. Starch yields of all cultivars ranged between 15 and 30 percent. Eleven of the 14 cultivars had starch yield less than 22 percent. However, three cultivars DM01-158-097 (28.4 percent), TIB4 (29.2 percent) and UKREDWHITE (28.8 percent) had high levels of starch. Our preliminary results showed that three cultivars might have potential for sweetpotato starch-based bio-ethanol production based on their starch yield.
Impacts
The broader impact of the biotechnology and genomics component is to discover and understand gene functions, and develop crop for high productivity and better human nutrition and health. These partial genomic and biotechnology results of sweetpotato and peanut have contributed to elucidate genome structure/organization and key physiological and biochemical functions. Peanut genome organization is not fully exploited because these transferable markers have aided in providing valuable DNA markers in peanut and allow for comparative mapping between peanut and soybean; thus, advancing peanut genomic research. The new RGH sequences obtained represent a resource of candidate genes and molecular markers for disease resistance in sweetpotato. The comparative molecular and physiological analyses were valuable in determining the time point for peak expressions-delayed with invitro-grown sweetpotato when compared to NFT, confirmation of storage root initiation between 10-14 days in NFT but delayed to 24-30 days post initial culture invitro, thus, revealing the same developmental pattern. The invitro microstorage root initiation methodologies could be utilized for assessment of physiological analyses and provide a quantitative method to measure specific transcripts within a cDNA such that sweetpotato in NFT. The outreach effort to Booker T. Washington High School afforded the opportunity to the students in the genetics class to investigate selected aspects within the field of plant tissue culture through applicability of lettuce media for the cloning of mustard and carrot plants using lettuce as control. Due to the unavailability of media specific to carrot and mustard, lettuce media was utilized for the set experiment. The students performed in-vitro germination of plantlets from seeds on MS media. The experiment showed 100 percent germination of all seeds on the MS media. On the shoot initiation media, lettuce produced greater number of calli, with about 75 percent calli production of the mustard and 50 percent calli production of carrot. On the root initiation media, all plants rooted, however, the rooting system of the carrot was not as strong as the lettuce and mustard.
Publications
- Wang, Y., B. Rosen, J. Scoffield, M. Egnin, D. Mortley, S. Steiner, D. R. Cook and G. He. 2008. Isolation and diversity analysis of resistance gene homologs (RGHs) in sweet potato. Theoretical and Applied Genetics (TAG-2007-0504). Submitted.
- Egnin, M., D. Mortley, E. F. Sanders, S. Traore, G. Gao, S. Jack, and T. Radwan. 2008. Comparative Gene Expression Profiling and the Physiological Role of t-Zeatin Riboside (ZR) Between In Vitro and Hydroponic-grown Sweetpotato During Storage Root Initiation and Enlargement. In Vitro Cell and Dev. Journal 44 (4): 355-356.
- Sanders, E. F., H. Guohao, L. Gong, M. Egnin and D. Morley. 2008. Transferability of Soybean (Glycine Max) SSR Markers in Peanut Genomic DNA (Arachis hypogaea L.). In Vitro Cell and Dev. Journal 44 (4): 356-357.
- Harrison, C.A., M. Egnin and S. Traore. 2008. Plant Tissue Culture Technique for the Secondary Classroom. In Vitro Cell and Dev. Journal 44 (4): 346.
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