DEVELOPMENT OF HYBRID AND DISEASE-RESISTANT RICE GERMPLASM FOR LOUISIANA

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 1015900
• Project Start Date: Apr 10, 2018
• Project End Date: Mar 31, 2022
• Program Code: [(N/A)]- (N/A)

Recipient Organization
LOUISIANA STATE UNIVERSITY
202 HIMES HALL
BATON ROUGE,LA 70803-0100

Performing Department
Rice Research Station

Non Technical Summary
Rice is a vital grain that contributes to food security for up to one-half of the world. In Louisiana, rice is an important economiccommodity with total gross farm value of 312million dollars in 2017. Louisiana rice provides employment and creates stablemilling, processing, and packaging industries that make substantial contributions to the economy of our state. In spite of thesesuccesses, economic margins for Louisiana rice producers are small due to high input costs for land preparation, fertilizers,pesticides, electrical and diesel pumping for water, harvest, drying, and storage of grain, and renting of land from secondarylandowners. Hybrid rice varieties can produce up to 15% more grain yield vs. inbred varieties in Louisiana and increase netreturn. The LSU AgCenter initiated a hybrid rice development program in 2009 to develop new germplasmwith high grain yield potential coupled with grain quality suitable for both internal and export markets. Development of newhybrid germplasm will be accomplished by standard field breeding methods that also exploit culture of rice cells in the laboratoryand use of DNA marker technology. The five research objectives for this Project aim to breed "foundation stocks" for hybridsadapted to Louisiana field environments, develop new hybrid combinations with viable economic potential, and create newhybrid seed production technologies adapted to U.S. conditions. Acquisition and development of new hybrid germplasm sourceswill be accomplished by cooperative agreements among public U.S. rice researchers and international institutes primarily in Asiaand South America. Successful completion of project goals is expected to create new hybrid germplasm that will increase grainyield, enhance grain quality over current hybrid varieties, and increase net economic return for Louisiana rice producers.

Animal Health Component

20%

Research Effort Categories

Basic

10%

Applied

20%

Developmental

70%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
201	1530	1081	100%

Knowledge Area
201 - Plant Genome, Genetics, and Genetic Mechanisms;

Subject Of Investigation
1530 - Rice;

Field Of Science
1081 - Breeding;

Keywords

rice

rice varieties

cultivar

grain

hybrid rice

Goals / Objectives
Create and/or identify male sterile (cytoplasmic A or environmental sensitive), restorer (R), and maintainer (B) lines adapted to Louisiana and southern U.S. environmental conditions.Develop elite hybrids through extensive test-crossing of lines developed in Objective 1 with emphasis on grain yield and quality, disease resistance, seedling vigor, earliness, ratoon potential, lodging resistance, environmental stability, and herbicide tolerance.Develop effective and economical hybrid seed production technology for Louisiana by modifying or optimizing available methods.Identify and utilize DNA markers to enhance grain quality and male sterility in selected elite inbred lines.Develop sheath blight-resistant rice breeding lines by conventional and molecular breeding methods.

Project Methods
Field plot research: Appropriate field plot methods including randomized complete block (RCBD) and augmented designs will beused for different field trials and experiments. Data collected will be entered into Excel spreadsheets and analyzed by SAS or other stistical software.Each year the following trials and plots will be planted at the Rice Research Station, Crowley, LA: F1 nursery, Observationaltestcross nursery, restorer (R) line development, maintainer (B) line development, male sterile (A and S) line development, fivecrossing blocks each consisting of ~ 120 entries planted every 10-14 days, micro and small-scale seed production plots. Up tofive off-station locations will be planted yearly with replicated yield trials of candidate hybrids along with appropriate commercialhybrids and inbreds. Anther culture will be used as needed at the Rice Research Station with certain material for rapid.development of homozygous lines. The Lajas winter nursery in Puerto Rice will be used for rapid selection and generationadvance of S, R, and B lines. Acquisition and evaluation ofnew germplasm will be accomplished by cooperative efforts with the University of Arkansas, other U.S. rice institutions, theUSDA rice germplasm collection, and international sources in China, the Philippines, and other countries. Each year anextensive crossing effort (up to 1,000 pollination events) will be carried out for development of new A, B, R, and S lines. HybridF1 material will be evaluated depending on objectives under appropriate field and/or greenhouse conditions. Anther culture andknown DNA molecular markers for traits, such as plant height, amylose content, and rice blast disease resistance, will be used forrapid generation advance and seed purity purposes.Data will be collected as needed from breeding lines and varieties for the following traits: date of emergence, date of heading,height, pollen fertility, panicle fertility, grain yield per plot, head rice and total rice milling yields, grain dimensions, percent chalkygrain, percent amylose, gelatinization temperature, percent lodging, disease and insect resistance, and response to varyingfertilizer rates. Data will be analyzed with corresponding and appropriate experimental designs using SAS v9.1 software.Evaluation:Success of the Project will be measured in terms of successful completion of the three primary research objectives. ForObjectives 1, this means development of male sterile A and S lines that exhibit desirable traits for pollen sterility/fertility, stigmaexsertion rates, high outcrossing rates, and high seed production rates for S lines. Yearly milestones will be evaluated foradvances in each type of germplasm in Objective 1. For Objective 2 this means development of appropriate hybrid combinationsthat produce seed yields and grain quality that approach or exceed current commercial hybrids grown in Louisiana. Recent trialsin 2018indicate that some of our new hybrid combination approach yield and quality values vs. some commercial hybrids.Multiple yield trials over years and locations during this Project will be needed to determine the potential of any new hybridcombination. For Objective 3, creation of a new seed production technology will be considered successful when hybrid seedproduction yields reach a commercial threshold of ~ 1,650 to 2,200 kg/h. We have recently identified some parentalcombinations in small field plots that meet or exceed this threshold.

Progress 10/01/19 to 09/30/20

Outputs
Target Audience:1. Louisiana rice producers and millers. 2. Louisiana, national, and international rice researchers. 3. National and international scientists involved in rice genetic improvement and disease resistance. Effort: Research results obtained from this Project disseminated through formal classroom instruction at LSU. Research results and practical impact transmitted throughout the course of this Project via extension and outreach to LSU AgCenter rice extension agents and Louisiana rice producers' meetings. Research progress also was reported at different regional and national scientific conferences. Changes/Problems:No changes or problems to report. What opportunities for training and professional development has the project provided?Three PhD graduate students are currently conducting research in the Hybrid Rice Breeding program and taking classes at Louisiana State University. How have the results been disseminated to communities of interest?Results from the research described in the five Objectives have been disseminated via various refereed and local LSU AgCenter publications, field day demonstrations, grower meetings, and LSU AgCenter research station newsletters, videos,and webpages. What do you plan to do during the next reporting period to accomplish the goals?1. Create and/or identify male sterile (cytoplasmic A or environmental sensitive S), restorer (R), and maintainer (B) lines adapted to Louisiana and southern U.S. environmental conditions. We will continue development and testing of new male sterile, restorer, and maintainer lines for improved performance and adaptability in multiple environments. 2. Develop elite hybrid germplasm and varieties through extensive test-crossing of lines developed in Objective 1 with emphasis on grain yield and quality, disease resistance, seedling vigor, earliness, ratoon potential, lodging resistance, environmental stability, and herbicide tolerance. We will evaluate new and advanced hybrid combinations for high yield potential, good grain quality, and improved grain appearance at multi-location sites in Louisiana. 3. Develop effective and economical hybrid seed production technology for Louisiana by modifying or optimizing available methods. We will evaluate seed treatment, planting date, seed density, and other novel approaches to maximizeseed production of new hybrid combinations. 4. Identify and utilize DNA markers to enhance grain quality and male sterility in selected elite inbred lines. We will validate selected markers and identify new candidate genomic regions for grain appearance and high milling yields in various elite lines and populations. We will use the validated markers to speed up development of new male sterile lines. 5. Develop sheath blight-resistant rice by conventional and molecular breeding methods. We will transfer sheath blight resistance from advanced selected lines to elite inbred cultivars and advanced hybrid experimentals by traditional breeding methods combined with trait-based molecular markers.

Impacts
What was accomplished under these goals? 1. Create and/or identify male sterile (cytoplasmic A or environmental sensitive), restorer (R), and maintainer (B) lines adapted to Louisiana and southern U.S. environmental conditions. Four new male sterile lines were developed that exhibited stable pollen sterility and good plant type, plant height, and maturity that are adapted to Louisiana field conditions. This material is currently being used in the development of new hybrid rice varieties for Louisiana and the southern United States. 2. Develop elite hybrids through extensive test-crossing of lines developed in Objective 1 with emphasis on grain yield and quality, disease resistance, seedling vigor, earliness, ratoon potential, lodging resistance, environmental stability, and herbicide tolerance. Using the male sterile lines from Objective 1, fournew hybrid combinations were identified in 2020that showed 15-30% yield advantage over elite commercial varieties grown in Louisiana. The new hybrids also produced high head-rice yields, low to moderate endosperm chalk, and moderate to good levels of resistance to the major microbial pathogens. 3. Develop effective and economical hybrid seed production technology for Louisiana by modifying or optimizing available methods. Rapid and effective hybrid seed production is a major time and space constraint for breeding pipelines in U.S. hybrid rice. We continue tooptimizeplot size and configuration, seed density, and PGR timing for maximum F1 seed production in the southernUnited States that accelerates germplasm development compared to previous breeding strategies. 4. Identify and utilize DNA markers to enhance grain quality and male sterility in selected elite inbred lines. We have identified several SNP-based DNA markers for grain chalk and thermosensitive genetic male sterility for U.S. adapted rice lines. This marker technology will greatly accelerate the timeline for development of U.S. hybrids with improved grain quality. 5. Develop sheath blight-resistant rice breeding lines by conventional and molecular breeding methods. We have identified one new elite inbred breeding linewith high levels of tolerance to sheath blight disease, good grain yield, good height, and plant type in 2020that will be evaluated in 2021Louisiana multi-location yield trials.

Publications

• Type: Journal Articles Status: Published Year Published: 2020 Citation: Camacho, J.R., S.D. Linscombe, E.P. Webster, and J.H. Oard. 2020. Inheritance of Resistance and Response of ProvisiaTM Rice to Quizalofop-P-1 Ethyl under U.S. Field Conditions. Weed Technology. 1-21. doi:10.1017/wet.2019.114.
• Type: Other Status: Published Year Published: 2020 Citation: Harrell, D., R. Zaunbrecher, S. Brown, E.P. Webster, J.P. Hebert, B. Wilson, Blake, K.A. Fontenot, J.H. Oard, D.E. Groth, M. Kongchum, and A. Famoso. 2020. Rice Varieties and Management Tips. Pub. 2270, pp. 27, LSU AgCenter.

Progress 10/01/18 to 09/30/19

Outputs
Target Audience:1. Louisiana rice producers and millers. 2. Louisiana, national, and international rice researchers. 3. National and international scientists involved in rice genetic improvement and disease resistance. Effort: Research results obtained from this Project disseminated through formal classroom instruction at LSU. Research results and practical impact transmitted throughout the course of this Project via extension and outreach to LSU AgCenter rice extension agents and Louisiana rice producers' meetings. Research progress also was reported at different regional and national scientific conferences. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Four PhD graduate students are currently conducting research in the Hybrid Rice Breeding program and taking classes at Louisiana State University. How have the results been disseminated to communities of interest?Results from the research described in the five Objectives have been disseminated via various refereed and local LSU AgCenter publications, field day demonstrations, grower meetings, and LSU AgCenter research station newsletters and webpage. Effort: Research results obtained from this Project disseminated through formal classroom instruction at LSU. Research results and practical impact transmitted throughout the course of this Project via extension and outreach to LSU AgCenter rice extension agents and Louisiana rice producers' meetings. Research progress also was reported at different regional and national scientific conferences. What do you plan to do during the next reporting period to accomplish the goals?1. Create and/or identify male sterile (cytoplasmic A or environmental sensitive S), restorer (R), and maintainer (B) lines adapted to Louisiana and southern U.S. environmental conditions. We will continue development and testing of new male sterile, restorer, and maintainer lines for improved performance and adaptability in multiple environments. 2. Develop elite hybrid germplasm and varieties through extensive test-crossing of lines developed in Objective 1 with emphasis on grain yield and quality, disease resistance, seedling vigor, earliness, ratoon potential, lodging resistance, environmental stability, and herbicide tolerance. We will evaluate new and advancedhybrid combinations for high yield potential, good grain quality, and improved grain appearance at multi-location sites in Louisiana. 3. Develop effective and economical hybrid seed production technology for Louisiana by modifying or optimizing available methods. We will evaluate seed treatment, planting date, seed density, and other novel approaches to optimize seed production of new hybrid combinations. 4. Identify and utilize DNA markers to enhance grain quality and male sterility in selected elite inbred lines. We will validate selected markers and identify new candidate genomic regions for grain appearance and high milling yields in various elite lines and populations. We will use the validated markers tospeed up development of new male sterile lines. 5. Develop sheath blight-resistant rice by conventional and molecular breeding methods. We will transfer sheath blight resistance from advanced selected lines to elite inbred cultivars and advanced hybrid experimentals by traditional breeding methods combined withtrait-based molecular markers.

Impacts
What was accomplished under these goals? 1. Create and/or identify male sterile (cytoplasmic A or environmental sensitive), restorer (R), and maintainer (B) lines adapted to Louisiana and southern U.S. environmental conditions. Three new male sterile lines weredeveloped that exhibited stable pollen sterility andgood plant type,plant height, and maturity that are adaptedto Louisiana field conditions. This material is currently being used in the development of new hybrid rice varieties for Louisiana and the southern United States. 2. Develop elite hybrids through extensive test-crossing of lines developed in Objective 1 with emphasis on grain yield and quality, disease resistance, seedling vigor, earliness, ratoon potential, lodging resistance, environmental stability, and herbicide tolerance. Using the male sterile lines from Objective 1, seven new hybrid combinations were identified in 2019 that showed 15-20% yield advantage over elite commercial varieties grown in Louisiana. The new hybrids also produced highhead-rice yields, low endosperm chalk, and moderate to good levels of resistance to the major microbial pathogens. 3. Develop effective and economical hybrid seed production technology for Louisiana by modifying or optimizing available methods. Rapid and effective hybrid seed production is a major time and space constraint for breeding pipelines in U.S. hybrid rice. We have optimized plot size and configuration, seed density, and PGR timing for maximum F1 seed production in the southern UnitedStates that accelerates germplasm development compared to previous breeding strategies. 4. Identify and utilize DNA markers to enhance grain quality and male sterility in selected elite inbred lines. We have identified several SNP-based DNA markers for grain chalk and thermosensitive genetic male sterility for U.S. adapted rice lines. This marker technology will greatly accelerate the timeline for development of U.S. hybrids with improved grain quality. 5. Develop sheath blight-resistant rice breeding lines by conventional and molecular breeding methods. We have identified twonew elite inbred breeding lines with high levels of tolerance to sheath blight disease, good grain yield, good height, and planttype in 2019that will be evaluated in 2020Louisiana multi-location yield trials.

Publications

• Type: Journal Articles Status: Published Year Published: 2019 Citation: De Guzman, C.T., S.D. Linscombe and J.H. Oard. 2019. Genetic analysis of environment-sensitive genic male sterile rice under US environments. Euphytica 215:39-46.
• Type: Book Chapters Status: Published Year Published: 2019 Citation: De Guzman, C.T. and J.H. Oard. 2019. Genetics and breeding system for cytoplasmic and genetic male sterility in rice. in Protecting Rice Grains in the Post-Genomic Era, InterOpen Limited Press, London. DOI: 10.5772/intechopen.85191.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Harrell, D., R. Zaunbrecher, S. Brown, E.P. Webster, J.P. Hebert, B.E. Wilson, K.A. Fontenot, J.H. Oard, D.E. Groth, M. Kongchum, and A. Famoso. 2018. Rice Varieties and Management Tips 2019. Pub. 2270, pp. 27, LSU AgCenter.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Camacho, J.R., S.D. Linscombe, Y. Sanabria, P.A. Mosquera, and J.H. Oard. 2019. Inheritance of Provisia" Rice Resistance to Quizalofop-P-Ethyl Under Laboratory and Greenhouse Environments. Euphytica 215:83. https://doi.org/10.1007/s10681-019-2407-4.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Famoso, A.N., D.L. Harrell, D.E. Groth, M. Kongchum, I. Wenefrida, J.H. Oard, R.E. Zaunbrecher, K.F. Bearb, C.A. Conner, G.J. Guidry, B. Angira, X. Sha, and S.D. Linscombe. 2019. Registration of CLJ01 Rice. J. Plant Reg. 13:143147.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Famoso, A.N., D.L. Harrell, D.E. Groth, E.P. Webster, J.H. Oard, R.E. Zaunbrecher, K.F. Bearb, C.A. Conner, G.J. Guidry, B. Angira, L.M. White, and S.D. Linscombe. 2019. Registration of PVL01 Rice. J. Plant Reg. (doi:10.3198/jpr2019.01.0002crc).
• Type: Journal Articles Status: Accepted Year Published: 2019 Citation: Angira, B., C.K. Addison, T. Cerioli, D.B. Rebong, D.R. Wang, N.S. Pumplin, J.H. Ham, J.H. Oard, S.D. Linscombe, and A.N. Famoso. 2019. Haplotype Characterization of the sd1 Semi-dwarf Gene in US Rice Germplasm. Plant Genome (accepted).
• Type: Journal Articles Status: Accepted Year Published: 2019 Citation: Camacho, J.R., S.D. Linscombe, E.P. Webster, and J.H. Oard. 2019. Inheritance of Resistance and Response of ProvisiaTM Rice to Quizalofop-P-1 Ethyl under U.S. Field Conditions. Weed Technology (accepted).

Progress 04/10/18 to 09/30/18

Outputs
Target Audience:1. Louisiana rice producers and millers. 2. Louisiana, national, and international rice researchers. 3. National and international scientists involved in rice genetic improvement and disease resistance. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Five PhD graduate students are currently conducting research in the Hybrid Rice Breeding program. How have the results been disseminated to communities of interest?Results from the research described in the five objectives have been disseminated via various refereed and local LSU AgCenter publications, field day demonstrations, grower meetings, LSU AgCenter research station newsletters, and webpage. Description of the LAH169 hybrid has been published in different online and hardcopy media outlets in 2018. What do you plan to do during the next reporting period to accomplish the goals?1. Create and/or identify male-sterile (cytoplasmic A or environmental sensitive S), restorer (R), and maintainer (B) lines adapted to Louisiana and southern U.S. environmental conditions. Develop and testnew male-sterile, restorer, and maintainer lines for improved performance and adaptability to different Louisiana environments. 2.Develop elite hybrid germplasm and varieties through extensive test-crossing of lines developed in Objective 1 with emphasis on grain yield and quality, disease resistance, seedling vigor, earliness, ratoon potential, lodging resistance, environmental stability, and herbicide tolerance. Evaluate multiple new testcross hybrid combinations for high yield potential, good grain quality, and improved grain appearance in multi-locations field trials. 3. Develop effective and economical hybrid seed production technology for Louisiana by modifying or optimizing available methods. Evaluate seed treatment and other novel approaches to optimize seed production of new hybrid combinations. 4. Identify and utilize DNA markers to enhance grain quality and male sterility in selected elite inbred lines. a. Identify new markers and validate candidate markers for grain appearance and high milling yields. b. Utilize markers to speed up advancement of selected male-sterile lines. 5. Develop sheath blight-resistant rice by conventional and molecular breeding methods. Transfer sheath blight resistance from advanced selected lines to elite inbred cultivars and advanced hybrid experimentals by traditional crossing approaches and use of trait-based molecular markers.

Impacts
What was accomplished under these goals? To provide additional options for sourthern U.S. rice producers, the LSU AgCenter released in March 2018 the first public hybrid rice variety. LAH169 is an earl-maturing, conventional long-grain hybridwith good yield potential and excellent grain quality. Additional candidates are in the breeding pipeline for future release.The developmentof LAH169 establishes a foundation for universityparticipation in the success of hybrid rice and underscores the value of public funding thatwill benefit the U.S. rice industry. 1. Create and/or identify male-sterile (cytoplasmic A or environmental sensitive), restorer (R), and maintainer (B) lines adapted to Louisiana and southern U.S. environmental conditions. A cytoplasmic orcms A line was developed with stable pollen sterility, good plant height, and seed production that is adapted to Louisiana field conditions. This material is currently being used in the development of new hybrid rice varieties for Louisiana and the southern United States. 2. Develop elite hybrids through extensive test-crossing of lines developed in Objective 1 with emphasis on grain yield and quality, disease resistance, seedling vigor, earliness, ratoon potential, lodging resistance, environmental stability, and herbicide tolerance. LAH169 is an early-maturing, conventionallong-grain hybrid releasedby the LSU AgCenter in March 2018. The combined yield of the main and ratoon cropwas 13,800lb/A across two Louisiana locations from 2015-2016 that represents a 20% yield advantage over the two most popular Louisiana conventional varieties. LAH169 produces desiredgrain dimensions, high head-rice yields, low endosperm chalk, and moderate to good levels of resistance to the major microbial pathogens. We expect LAH169 to provide a viable economic option for Louisiana rice producers. 3. Develop effective and economical hybrid seed production technology for Louisiana by modifying or optimizing available methods. Rapid and effective hybrid seed production is a major time and space constraint for breeding pipelines in U.S. hybrid rice. We haveoptimized plotsizeand configuration, seed density, and PGR timing for maximum F1 seed production in the southern U.S. that accelerates germplasm development compared to previous breeding strategies. 4.Identify and utilize DNA markers to enhance grain quality and male sterility in selected elite inbred lines. We have identified several SNP-based DNA markers for grain chalk and thermosensitive genetic male sterility for U.S.-adapted rice lines. This marker technology will greatly accelerate the timeline for development of U.S. hybrids with improved grain quality. 5. Develop sheath blight-resistant rice breeding lines by conventional and molecular breeding methods. We have identified five elite inbred breeding lines with high levels of tolerance to sheath blight disease, good height, and plant type in 2018 that will be evaluated in 2019 Louisiana multi-location yield trials.

Publications

• Type: Other Status: Published Year Published: 2018 Citation: Harrell, D., Brown, S., Webster, E., Hebert, J., Wilson, B., Zaunbrecher, R., Fontenot, K., Oard, J., Groth, D., Kongchum, M., and Famoso, A. 2018. Rice Varieties and Management Tips. LSU AgCenter Pub. 2270.
• Type: Journal Articles Status: Submitted Year Published: 2018 Citation: Jose R. Camacho, Steve D. Linscombe, Yamid Sanabria, Paola A. Mosquera, and James H. Oard. 2018. Inheritance of Provisia" Rice Resistance to Quizalofop-P-Ethyl Under Laboratory and Greenhouse Environments. Euphytica Submission ID: EUPH-D-18-00709.
• Type: Journal Articles Status: Submitted Year Published: 2018 Citation: Christian Torres De Guzman, Steven D. Linscombe, and James H. Oard. 2018. Genetic Analysis of Environmentally-Sensitive Male Sterile Rice under US Environments. Euphytica Submission ID: EUPH-D-18-00625.