ESTABLISHMENT AND APPLICATION OF A VIRUS-FREE SWEETPOTATO PROGRAM FOR LIMITED RESOURCE FARMERS IN MISSISSIPPI

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: OTHER GRANTS
• Reporting Frequency: Annual
• Accession No.: 1007771
• Grant No.: 2015-38821-24345
• Cumulative Award Amt.: $248,719.00
• Proposal No.: 2015-06248
• Project Start Date: Sep 1, 2015
• Project End Date: Aug 14, 2020
• Grant Year: 2015
• Program Code: [EWE]- Extension Project

Project Director
Meng, Y.

Recipient Organization
ALCORN STATE UNIVERSITY
(N/A)
ALCORN STATE,MS 39096

Performing Department
(N/A)

Non Technical Summary
Sweetpotato (Ipomoea batatas L.) belongs to Convolvulaceae family, is an important crop for food security. As one of the top three vegetable crops grown in Mississippi, one major limitation to sweetpotato production is the cumulative effect of virus infection causing cultivar decline and yield losses. Technology such as meristem-tip culture can provide farmers with healthy propagating materials that are free of detectable viruses. However, it has not been well practiced in Mississippi, particularly in the small farms. The overall goal of this project is to establish and employ a virus-free sweetpotato program in Mississippi for limited resource farmers, aiming at increasing the sweetpotato yield, quality and investment return for small holders. Specifically, the project seeks to: (1) develop an integrated, meristem-tip culture-based protocol for the production of virus-free sweetpotato; (2) integrate virus detection into virus-indexed sweetpotato production and apply major virus diagnosis to sweetpotato production by small holders in Mississippi; (3) establish an effective propagation and delivery system of virus-indexed sweetpotato plants to limited resource farmers; (4) provide training modules and technical support to diverse groups of agricultural research and extension services at Alcorn State University. Successful execution of this project will significantly boost quality of life and environment in the underserved communities and enhance research, education and extension capacities of Alcorn State University.

Animal Health Component

(N/A)

Research Effort Categories

Basic

100%

Applied

(N/A)

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
204	1450	1101	100%

Knowledge Area
204 - Plant Product Quality and Utility (Preharvest);

Subject Of Investigation
1450 - Sweet potato;

Field Of Science
1101 - Virology;

Keywords

sweetpotato

virus infection

meristem-tip culture

"seed" program

limited resource farmers

Goals / Objectives
Major goals: The overall goal of this project is to establish and employ a virus-free sweetpotato program in Mississippi for limited resource farmers, aiming at increasing the sweetpotato yield, quality and investment return for small holders. Specifically, the project seeks to:To develop an integrated, meristem-tip culture-based protocol for the production of virus-free sweetpotato;To integrate virus detection into virus-indexed sweetpotato production and apply major virus diagnosis to sweetpotato production by small holders in Mississippi;To establish an effective propagation and delivery system of virus-indexed sweetpotato plants to limited resource farmers;To provide training modules and technical support to diverse groups of agricultural research and extension services at Alcorn State University.

Project Methods
Objective 1: To develop an integrated, meristem-tip culture-based protocol for the production of virus-free sweetpotatoWe will use the meristem shoot tip tissue culture procedure developed at the project collaborator's lab at the U.S. Vegetable Laboratory, USDA-ARS in Charleston, South Carolina (Ling et al., 2010). Briefly, after sterilization in a chlorine bleach solution, aseptically isolated meristem tips (~0.5 mm) will be placed onto Petri dishes (10 cm in diameter) containing Meristem Shoot Initiation Medium (1 packet of SIGMA MS salts, 30 g sucrose, 4 mL of 500 mg/mL thiamine HC, 0.1 g of Myo-inositol, 2.5 g Phytagel/L and 500 mg/mL 6-benzyl-aminopurine/L, adjusted to pH 5.8). The excised meristem shoot tips in dishes are placed in a tissue culture room with 16h/day lighting and 25-30 °C. Newly developed shoots will be transferred to Phyta trays in Regeneration Medium (1 packet of GIBCO salts, 5 mL GIBCO vitamins, 50 g sucrose and 2.5 g Phytagel/L) for further shoot growth and root development. Plantlets with roots are then transferred to test tubes with Gibco MS Medium (1 packet of GIBCO salts, 5 mL GIBCO vitamins, 30 g sucrose and 2.5 g Phytagel/L).For quality control purpose, newly regenerated plantlets will be tested for the presence of viruses with the detection methods described below. Plantlets with no detectable viruses using ELISA and PCR will be saved as stock for further increase. The virus-status of such plantlets will be further confirmed through graft indexing on Ipomoea setosa plants. In this study, we will develop virus-tested 'Beauregard', 'Centennial', 'Jewel', 'O'Henry and 'Evangeline' seed stocks and maintain the variety purity via tissue culture. Beauregard has been shown to be the main variety planted in Mississippi in 2012 (NCAGR, 2014) and other major varieties are also included for propagation.Objective 2: To integrate virus detection into virus-indexed sweetpotato production and apply major virus diagnosis to sweetpotato production by small holders in MississippiMore than 30 viruses from 9 families have been isolated and described on sweetpotato globally (Clark et al., 2012). Many antibodies to these viruses are available from Agdia Company that makes the protein-based ELISA assay convenient as a low-cost, large-scale test to those viruses. Alternatively, the nucleic acid-based polymerase chain reaction (PCR) and reverse-transcription PCR (RT-PCR) will be used to achieve higher sensitivity and confirmation at the genomic level of viral species and strains. Primers targeting to conserved regions of the know sweetpotato viruses will be used for this nucleic acid based detection and the amplified product will be subject to sequencing to gain further genetic information (Ling et. al., 2010). Multiplex PCR will be developed for the simultaneous detection of multiple viruses in important samples. In addition to sweetpotato, other Ipomoea species such as Ipomoea setosa that can cause visible symptoms for many of sweetpotato viruses will be used as indicator plant in grafting experiment to facilitate detection.We plan to use these methods on the tip culture plantlets, stocks, green house and station propagules to detect and remove infected materials. Further, the diagnosis and detection methods will be used to test samples collected from farms and extension fields to gain additional information on major viral diseases affecting small farm sweetpotato production in Mississippi.Objective 3: To establish an effective propagation and delivery system of virus-indexed sweetpotato plants to limited resource farmersA three-component system that comprises of tissue culture lab, propagating nursery and end farm users will be established for the propagation of virus-indexed sweetpotato seeds and delivery to small farm holders.Original stock plants (G0): Original virus-free sweetpotato stock plants that are obtained from heat treatment and meristem-tip culture will be checked routinely to be free of detectable viruses. The maintenance and propagation of the original stock plants will be done in tissue culture lab located in the Agriculture Extension Building at ASU. Samples will be collected and labeled for detection at the plant pathology lab in Biotechnology Center at ASU.Indexed original plants (G1): Tissue culture plants with undetectable viruses will be increased by nodal propagation from the original stock plants G0. Then the new plantlets will be planted in a net-cage placed in the greenhouse at Alcorn State University. The plants in the greenhouses will be increased by vine cuttings through the winter (G1). Plant leaf samples will be collected and tested in batch for G1 plants before they are delivered to G2 growers.Commercial "Seed" sweetpotato (G2): G1 plants will be used as a source of vine cuttings to certified commercial seed growers to produce G2 seeds for sale to table-stock growers. Propagation of productive "seed" sweetpotato from the original plants is carried out in fields where sweetpotato has not been grown for several years before and where no other sweetpotato cultivars are planted within 500 meters. During propagation, trained personnel will inspect the production field and discard plants with suspicious symptoms. At the same time, virus-indexed sweetpotato "seeds" will be made available to demo-farms for yield evaluation and demonstration to small holders. After cycles of propagation, a great quantity of virus-free "seed" sweetpotato (vines or storage roots) will be provided to small holders for commercial production.Food sweetpotato and production sweetpotato (G3): Generally, the commercial sweetpotato G2 "seeds" are used for food sweetpotato production. Or if necessary, G2 plants from commercial seed growers can be used by small holders to further increase sweetpotato "seeds" (G3) to cut cost and meet growth season requirement. The project personnel will establish and maintain training sessions to small holders during production seasons.Objective 4: To provide training modules and technical support to diverse groups of agricultural research and extension services at Alcorn State UniversitySummer workshops on virus-free sweetpotato demonstration and virus disease diagnostics will be held at ASU, lab and field-training sessions will also be included. The workshop is set up to train extension agents, farmers and students at ASU. The workshop will include a combination of lectures and laboratory exercises. The lectures will include topics related to tissue culture techniques, plant virus disease diagnosis as well as plant health management. Attendees will tour the tissue culture production facility and ASU sweetpotato demonstration field to promote interactions between university researchers and small holders. The plant virus laboratory exercises will provide hands-on training in different aspects of conventional and modern diagnostic and identification techniques. Catalogued samples from research effort and specimens procured from professional associations such as American Phyopathological Society (APS) will be utilized to allow audiences to gain hands-on experience on viral disease diagnosis. Occasionally, experts in related fields will be invited to present seminars on various topics to broaden the learning experience. Advanced trainings for extension specialists and graduate students will include plant tissue culture techniques, viral symptom observation of various plants maintained in the lab, ELISA assay, plant nucleic acid extraction, PCR and RT-PCR, DNA sequencing and bioinformatics analysis.Extension publications will be developed in print such as fact sheets, timely information, and bulletins on important plant pathogens. In addition, flyers will be printed for in filed distribution to limited resource farmers where outreach is hindered by lack of communication means.

Progress 09/01/15 to 08/31/20

Outputs
Target Audience:During this reporting period, the target audiences were ASU research scientists, undergraduate and graduate students, small farmers and extension specialists. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?During the execution of this project, the students, lab technician, extension agents were trained with better skills and knowledge related with plant tissue culture techniques, disease diagnosis techniques and field practices. Two courses related with tissue culture and genetic engineering techniques were developed at undergraduate and graduate class levels at Alcorn State University and Hinds Community College. Flyers on plant disease management by applying modern biotechniques together with on-site demonstration have educated small farmers with raised awareness of viral disease knowledge based IPM for better environment protection. A close network among scientists, extension agents, farmers' associations and farmers greatly motivate the movement of biotechniques from research labs to the field practices, all the personnel mentioned above are benefit for the training and professional development. How have the results been disseminated to communities of interest?The research data were presented to several scientific meetings regionally or nationally, and stimulated exciting discussions for further collaborations. Posters, printed flyers and oral presentations during field days and workshops have direct impacted farmers knowledge base and raised discussion on how to integrated agricultural biotechnology into sweetpotato production practices. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? In general, we haveaccoplished the major goals of this project.The procedure to remove the sweetpotato viruses with a combination of the meristem shoot tip tissue culture and thermotherapy was optimized in our lab; Optimized PCR protocols were developed to detect the existence of the four RNA viruses and one DNA virus for sweetpotato;Samples provided by Mississippi farmers were used to conduct virus detection and virus removal.Annual field practice to evaluate the performance of virus-tested sweetpotato was conducted at ASU field; Virus-free sweetpotato field demonstration and disease diagnosis were held at ASU field day and related workshops.

Publications

• Type: Journal Articles Status: Published Year Published: 2020 Citation: Meng Y, Zhang C and Njiti V (2020). Virus elimination in sweetpotato: from meristem-tip culture to storage roots production: a review. Adv Agri Tech Plant Sciences 3(1): 180048
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Emmanuella Nnuji-John, Chimaka Udedibor, Yan Meng, Chunquan Zhang, Victor Njiti (2020). Virus-free sweetpotato: from meristem tip culture to storage roots production. 84th Annual Mississippi Academy of Sciences Meeting, Biloxi, Mississippi
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Aundrea Coleman, Toni-Ann Nelson, Chunquan Zhang, Victor Njiti and Yan Meng (2020). Genetically engineered sweetpotato: a novel approach to improve sweet potato viral resistance. 84th Annual Mississippi Academy of Sciences Meeting, Biloxi, Mississippi
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Toni-Ann Nelson, Aundrea Coleman, Yan Meng. Development of efficient plant regeneration protocols from diverse genotypes of sweetpotato (Ipomoea batatas (L. Lam) (2020). 84th Annual Mississippi Academy of Sciences Meeting, Biloxi, Mississippi
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Myshawn Smith, Daniela F. Adjaye, Yan Meng, Victor Njiti, Chunquan Zhang Sweet potato polyphenol oxidase molecular study and its potential as CRISPR/Cas9 target for genome editing (2020). 84th Annual Mississippi Academy of Sciences Meeting, Biloxi, Mississippi
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Virus elimination in sweetpotato through meristem tip culture and thermotherapy (2020). Chimaka Udedibor, Emmanuella Nnuji-John, Yan Meng. 84th Annual Mississippi Academy of Sciences Meeting, Biloxi, Mississippi
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Andreya Dupree, Daniela Adjaye, Yan Meng, Victor Njiti, Chunquan Zhang (2020). Characterization of an infectious poplar mosaic virus CDNA clone. 84th Annual Mississippi Academy of Sciences Meeting, Biloxi, Mississippi

Progress 09/01/15 to 08/14/20

Outputs
Target Audience:During this reporting period, the target audiences were ASU research scientists, undergraduate and graduate students, small farmers and extension specialists. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?During the execution of this project, the students, lab technician, extension agents were trained with better skills and knowledge related with plant tissue culture techniques, disease diagnosis techniques and field practices. Two courses related with tissue culture and genetic engineering techniques were developed at undergraduate and graduate class levels at Alcorn State University and Hinds Community College. Flyers on plant disease management by applying modern biotechniques together with on-site demonstration have educated small farmers with raised awareness of viral disease knowledge based IPM for better environment protection. A close network among scientists, extension agents, farmers' associations and farmers greatly motivate the movement of biotechniques from research labs to the field practices, all the personnel mentioned above are benefit for the training and professional development. How have the results been disseminated to communities of interest?The research data were presented to several scientific meetings regionally or nationally, and stimulated exciting discussions for further collaborations. Posters, printed flyers and oral presentations during field days and workshops have direct impacted farmers knowledge base and raised discussion on how to integrated agricultural biotechnology into sweetpotato production practices. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? In general, we haveaccoplished the major goals of this project.The procedure to remove the sweetpotato viruses with a combination of the meristem shoot tip tissue culture and thermotherapy was optimized in our lab; Optimized PCR protocols were developed to detect the existence of the four RNA viruses and one DNA virus for sweetpotato;Samples provided by Mississippi farmers were used to conduct virus detection and virus removal.Annual field practice to evaluate the performance of virus-tested sweetpotato was conducted at ASU field; Virus-free sweetpotato field demonstration and disease diagnosis were held at ASU field day and related workshops.

Publications

• Type: Journal Articles Status: Published Year Published: 2020 Citation: Meng Y, Zhang C and Njiti V (2020). Virus elimination in sweetpotato: from meristem-tip culture to storage roots production: a review. Adv Agri Tech Plant Sciences 3(1): 180048
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Virus elimination in sweetpotato through meristem tip culture and thermotherapy (2020). Chimaka Udedibor, Emmanuella Nnuji-John, Yan Meng. 84th Annual Mississippi Academy of Sciences Meeting, Biloxi, Mississippi
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Andreya Dupree, Daniela Adjaye, Yan Meng, Victor Njiti, Chunquan Zhang (2020). Characterization of an infectious poplar mosaic virus CDNA clone. 84th Annual Mississippi Academy of Sciences Meeting, Biloxi, Mississippi
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Emmanuella Nnuji-John, Chimaka Udedibor, Yan Meng, Chunquan Zhang, Victor Njiti (2020). Virus-free sweetpotato: from meristem tip culture to storage roots production. 84th Annual Mississippi Academy of Sciences Meeting, Biloxi, Mississippi
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Aundrea Coleman, Toni-Ann Nelson, Chunquan Zhang, Victor Njiti and Yan Meng (2020). Genetically engineered sweetpotato: a novel approach to improve sweet potato viral resistance. 84th Annual Mississippi Academy of Sciences Meeting, Biloxi, Mississippi
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Toni-Ann Nelson, Aundrea Coleman, Yan Meng. Development of efficient plant regeneration protocols from diverse genotypes of sweetpotato (Ipomoea batatas (L. Lam) (2020). 84th Annual Mississippi Academy of Sciences Meeting, Biloxi, Mississippi
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Myshawn Smith, Daniela F. Adjaye, Yan Meng, Victor Njiti, Chunquan Zhang Sweet potato polyphenol oxidase molecular study and its potential as CRISPR/Cas9 target for genome editing (2020). 84th Annual Mississippi Academy of Sciences Meeting, Biloxi, Mississippi

Progress 09/01/18 to 08/31/19

Outputs
Target Audience:During this reporting period, the target audiences were ASU research scientists, undergraduate and graduate students, small farmers and extension specialists. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?During the execution of this project, the students, lab technician, extension agents were trained with better skills and knowledge related with plant tissue culture techniques, disease diagnosis techniques and field practices. Two courses related with tissue culture and genetic engineering techniques were developed at undergraduate and graduate class levels at Alcorn State University and Hinds Community College. Flyers on plant disease management by applying modern biotechniques together with on-site demonstration have educated small farmers with raised awareness of viral disease knowledge based IPM for better environment protection. A close network among scientists, extension agents, farmers' associations and farmers greatly motivate the movement of biotechniques from research labs to the field practices, all the personnel mentioned above are benefit for the training and professional development. How have the results been disseminated to communities of interest?The research data were presented to several scientific meetings regionally or nationally, and stimulated exciting discussions for further collaborations. Posters, printed flyers and oral presentations during field days and workshops have direct impacted farmers knowledge base and raised discussion on how to integrated agricultural biotechnology into sweetpotato production practices. What do you plan to do during the next reporting period to accomplish the goals?Our next goals will be focusing on goal 3 and 4. Especially, to establish an effective propagation and delivery system of virus indexed sweetpotato plants to limited resource farmers and to provide training modules and technical support to diverse groups of agricultural research and extension services. A netted greenhouse of Alcorn Agriculture Station at Mound Bayou, MS is constructed to serve for this purposes. The collaboration with Hinds Community College will further provide the virus free sweetpotato seeds to local farmers for increasing their farming income. PI, Co-PIs and collaborators will continue to arrange meetings to discuss plans for execution of the goals.

Impacts
What was accomplished under these goals? The procedure to remove the sweetpotato viruses with a combination of the meristem shoot tip tissue culture and thermotherapy was optimized in our lab. Totally 24 lines of sweetpotato, for purposes of delivery to farmers or for breeding, have been collected from USDA, Louisiana State University and Mississippi State University. Once lines being received, optimized PCR protocols were used to detect the existence of the four RNA viruses and one DNA virus. Annual field practice to evaluate the performance of virus-tested sweetpotato was conducted at ASU field; leaf and root samples were harvested and used for virus detection and analysis. Virus-free sweetpotato field demonstration and disease diagnosis were held at ASU field day and related workshops.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: 1.Yan Meng, Faith Iseguede, Chunquan Zhang, Victor Njiti (2019). Establish and application of a virus-free sweetpotato program for limited resource farmers in Mississippi. 2019 American Society for Horticultural Science Annual Conference, Las Vegas, Nevada
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: 2.Faith Iseguede, Yan Meng (2019). Biotechnological approaches for improving sweetpotato viral disease resistance in Mississippi. 34th Annual Career Fair & Training Conference, Kansas
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: 3.Faith Iseguede, Yan Meng, Chunquan Zhang, Victor Njiti (2019). Elimination and detection of viruses in meristem-derived plantlets of sweetpotato in Mississippi. 19th Biennial research Symposium for Association of 1890 Research Directors, Jacksonville, Florida
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: 4.Faith Iseguede, Yan Meng, Chunquan Zhang, Victor Njiti (2019). Using biotechnological approaches for improving sweetpotato viral disease resistance. 83rd Annual Mississippi Academy of Sciences Meeting, Hattiesburg, MS
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: 5.Faith Iseguede, Yan Meng, Chunquan Zhang, Victor Njiti (2019). Biotechnological approaches for improving sweetpotato viral disease resistance in Mississippi. National Sweetpotato Collaborator Group meeting, Birmingham, AL
• Type: Theses/Dissertations Status: Published Year Published: 2019 Citation: 6. Faith Iseguede (2019). Elimination of viruses in sweetpotato (Ipomoea Batatas) by meristem-tip culture and thermotherapy

Progress 09/01/17 to 08/31/18

Outputs
Target Audience:During this reporting period, the target audiences were ASU research scientists, undergraduate and graduate students, small farmers and extension specialists. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?During the execution of this project, the students, lab technician, extension agents were trained with better skills and knowledge related with plant tissue culture techniques, disease diagnosis techniques and field practices. Two courses related with tissue culture and genetic engineering techniques were developed at undergraduate and graduate class levels. Flyers on plant disease management by applying modern biotechniques together with on-site demonstration and presentation have educated small farmers raising awareness of viral disease knowledge based IPM for better environment protection. A close networking among scientists, extension agents, farmers' associations and farmers greatly motivate the movement of biotechniques from research labs to the field practices, all the personnel mentioned above are benefit for the training and professional, development. How have the results been disseminated to communities of interest?The results were presented to several scientific meetings regionally or nationally and stimulated exciting discussions for further collaborations. Posters, printed flyers and oral presentations during field days and workshops have direct impacted farmers knowledge base and raise discussion on how to integrated agricultural biotechnology into sweetpotato production practices. What do you plan to do during the next reporting period to accomplish the goals?Our next goals will be focusing on goal 3 and 4. Especially, to establish an effective propagation and delivery system of virus-indexed sweetpotato plants to limited resource farmers and to provide training modules and technical support to diverse groups of agricultural research and extension services. A netted greenhouse of Alcorn Agriculture Station at Mound Bayou, MS is in construction to serve for this purposes. PI, Co-PIs and collaborators will continue to arrange meetings to discuss plans for execution of the goals.

Impacts
What was accomplished under these goals? The procedure to remove the sweetpotato viruses with a combination of the meristem shoot tip tissue culture and heat-treatment was successfully developed in our lab. Totally 16 lines of sweetpotato, for purposes of delivery to farmers or for breeding, have been collected from USDA, Louisiana State University and Mississippi State University. Lines infected with viruses were done or are processing with treatments for viruses removal. Protocols for detecting viruses from the tissue culture plantlets were developed and adopted. Original virus-free sweetpotato stock plants that are obtained from meristem-tip culture and heat treatment are checked routinely to be free of detectable viruses. Field practices to evaluate the performance of virus-tested sweetpotato were conducted at ASU field, roots harvested from virus-tested plants and from non-treated plants have been compared; Leaf, stem and root samples have been collected and conducted for virus-detection. Virus-free sweetpotato demonstration and virus disease diagnostics were held at ASU field day and workshop. Presentations about virus-free sweetpotato lab protocol and field practices were giving by PI and Co-PIs.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Kyler Holmes, Chunquan Zhang, Yan Meng (2018). Analysis of full-length infectious genomic cDNA clones of SPFMV and SPLCV and exploiting the approaches of biotechnology in sweetpotato for virus diseases resistance, 5th Mcbios, Starkville, MS
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Kyler Holmes, Yan Meng and Chunquan Zhang (2018). Development of biotechnological approaches for improving sweetpotato with multiple viral disease resistance. Mississippi Academy Society meeting, Hattiesburg, MS.
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Daniela F. Adjaye, Yan Meng, Victor Njiti, Chunquan Zhang (2018). Whole genome sequencing and the construction of infectious Poplar Mosaic Virus Clones. 4th Annual Centers for Research Excellence Symposium, Alcorn State University
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Faith O. Iseguede, Naquan Thomas and Yan Meng (2018). Virus removal of sweetpotato crop using meristem-tip culture technique. 4th Annual Centers for Research Excellence Symposium, Alcorn State University
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Kyler Holmes, Yan Meng, Chunquan Zhang and Victor Njiti (2018). Utilization of biotechnological approaches for improving sweetpotato viral disease resistance. 4th Annual Centers for Research Excellence Symposium, Alcorn State University
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Kyler Holmes, Yan Meng, Chunquan Zhang and Victor Njiti (2018). Exploiting the use of biotechnology for improving sweetpotato virus disease resistance. National sweetpotato collaborator group meeting, Wilmington, NC
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Faith Iseguede, Victor Njiti, Chunquan Zhang, Yan Meng (2018). Establishment and application of a virus free sweetpotato program for limited-resource farmers in Mississippi, USDA/NIFA PD meeting, Washington DC.
• Type: Theses/Dissertations Status: Accepted Year Published: 2018 Citation: Kyler Holmes (2018). Development of biotechnological approaches for improving sweetpotato with multiple viral disease resistance, Master Degree Thesis, Alcorn State University.

Progress 09/01/16 to 08/31/17

Outputs
Target Audience:During this reporting period, the target audiences were ASU research scientists, undergraduate and graduate students, small farmers and extension specialists. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?During the execution of this project, the students, lab technician, extension agents were trained with better skills and knowledge related with plant tissue culture techniques, disease diagnosis techniques and field practices. Two courses related with tissue culture and genetic engineering techniques were developed at undergraduate and graduate class levels. Flyers on plant disease management by applying modern biotechniques together with on-site demonstration and presentation have educated small farmers raising awareness of viral disease knowledge based IPM for better environment protection. A close networking among scientists, extension agents, farmers' associations and farmers greatly motivate the movement of biotechniques from research labs to the field practices, all the personnel mentioned above are benefit for the training and professional, development. How have the results been disseminated to communities of interest?The results were presented to several scientific meetings regionally or notionally and stimulated exciting discussions for further collaborations. Posters, printed flyers and oral presentations during field days and workshops have direct impacted farmers knowledge base and raise discussion on how to integrated agricultural biotechnology into sweetpotato production practices. What do you plan to do during the next reporting period to accomplish the goals?Our next goals to be acocoplished will be 3 and 4. Especially, to establish an effective propagation and delivery system of virus-indexed sweetpotato plants to limited resource farmers and to provide training modules and technical support to diverse groups of agricultural research and extension services. PI, Co-PIs and collaborators will continue to arrange meetings to discuss plans for execution of the goals.

Impacts
What was accomplished under these goals? We continue to optimize the meristem shoot tip tissue culture procedure developed at the project collaborator's lab at the U.S. Vegetable Laboratory, USDA-ARS in Charleston, South Carolina (Ling et al., 2010). Totally 13 lines of sweetpotato, for purposes of delivery to farmers or for breeding, have been collected from USDA, Louisiana State University or lines bred at Alcorn State University. Lines infected with viruses were done or are processing with treatments for viruses removal. A combination of meristem-tip culture technique and heat-treatment was used to remove the tested-viruses; if the tested virus still could not be effectively removed, a second round of the same treatments on the previously treated plants would be used to eventually remove the viruses. Protocols for detecting viruses from the tissue culture plantlets were developed. Original virus-free sweetpotato stock plants that are obtained from meristem-tip culture and heat treatment are checked routinely to be free of detectable viruses. We continue to conduct field practices to evaluate the performance of virus-tested sweetpotato, roots harvested from virus-tested plants and from non-treated plants have been compared; Leaf, stem and root samples have been collected and conducted for virus-detection. Virus-free sweetpotato demonstration and virus disease diagnostics were held at ASU field day and workshop. Presentations about virus-free sweetpotato lab protocol and field practices were giving by PI and Co-PIs.

Publications

• Type: Conference Papers and Presentations Status: Accepted Year Published: 2017 Citation: 1. Kyler Holmes, Yan Meng (2017). Development of a virus-free sweetpotato program for limited-resource farmers in Mississippi. Mississippi Academy of Sciences, Hattiesburg, Mississippi 2. Kyler Holmes, Yan Meng (2017). Development and management of sweetpotato virus disease resistance through biotechnology approaches. Alcorn State University Research Symposium, Lorman, Mississippi 3. Yan Meng, David Henderson, Kyler Holmes, Rita Okoro, Victor Njiti, Chunquan Zhang (2016). Development of transgenic sweetpotato with multiple virus resistance in US. American Phytopathology Society, July 2016, Tampa, Florida. 4. Landrick Akrong, Yan Meng, Victor Njiti, Christopher Clark, Chunquan Zhang (2016). Construction of full-length genomic cDNA clones of Sweet potato feathery mottle virus. Journal of the Mississippi Academy of Sciences 61(1): 25 5. David Henderson, Chunquan Zhang, Victor Njiti, Yan Meng (2016) Development of transgenic sweetpotato with multiple virus resistance in US. Journal of the Mississippi Academy of Sciences 61(1): 27

Progress 09/01/15 to 08/31/16

Outputs
Target Audience:Scientists, undergraduate and graduate Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?During the execution of this project, the students, lab technician, extension agents were trained with better skills and knowledge related with plant tissue culture techniques, disease diagnosis techniques and field practices. Flyers on plant disease management by applying modern biotechniques together with on-site demonstration and presentation have educated small farmers raising awareness of viral disease knowledge based IPM for better environment protection. A close networking among scientists, extension agents, farmers' associations and farmers greatly motivate the movement of biotechniques from research labs to the field practices, all the personnel mentioned above are benefit for the training and professional development. How have the results been disseminated to communities of interest?The result was presented to annual national sweetpotato growers and scientists meeting and stimulated exciting discussions for further collaborations. Printed flyers and oral presentations during field days have direct impacted farmers knowledge base and raise discussion on how to integrated agricultural biotechnology into sweetpotato production practices. What do you plan to do during the next reporting period to accomplish the goals?Continue to work on goals 2, 3 and 4. Especially, to establish an effective propagation and delivery system of virus-indexed sweetpotato plants to limited resource farmers will be our next year's main focus. PI, Co-PIs and collaborators continue to arrange face-face meeting and teleconference meetings to discuss plans for execution of the goals.

Impacts
What was accomplished under these goals? We modified the meristem shoot tip tissue culture procedure developed at the project collaborator's lab at the U.S. Vegetable Laboratory, USDA-ARS in Charleston, South Carolina (Ling et al., 2010). Totally 12 lines of sweetpotato, for purposes of delivery to farmers or for breeding, have been collected from USDA, Louisiana State University or lines bred by co-PI Dr. Njiti at Alcorn State University. All 13 lines were done or are processing with meristem tip culture for removing the viruses. RT-PCR was used to detect the tissue culture plantlets for Sweet Potato Leaf Curl Virus and Feathery Sweet Potato Mottle Virus. Protocols were developed according to (Li et al., 2012) and (Li et al., 2004). Original virus-free sweetpotato stock plants that are obtained from heat treatment and meristem-tip culture will be checked routinely to be free of detectable viruses. The original stock plants are being maintained and propagated in tissue culture lab located in the Agriculture Extension Building at ASU. Samples are being collected and labeled for detection at the plant pathology lab in Biotechnology Center at ASU. We have been conducting field practices for virus-free sweetpotato since 2015, roots harvested from virus-tested plants and from non-treated plants have been compared and leaf samples have been collected for virus-detection. Virus-free sweetpotato demonstration and virus disease diagnostics were held at ASU field day on August 25 2016. Dr. Yan Meng gave a presentation about virus-free sweetpotato lab protocol and field practices. Farmers showed great interests on the future collaboration on Nursery Company.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: 1. Akrong, L, Meng Y, Njiti V, Clark C and Zhang C (2016). Construction of full-length genomic cDNA clones of Sweet potato feathery mottle virus. Journal of the Mississippi Academy of Sciences 61(1): 25 2. Henderson D, Zhang C, Njiti V and Meng Y (2016) Development of transgenic sweetpotato with multiple virus resistance in US. Journal of the Mississippi Academy of Sciences 61(1): 27