2004 Annual Report 1.What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? Sugarbeet (Beta vulgaris) accounts for about 55% of the domestic sucrose production in USA. Sugarbeet has a narrow germplasm base having been selected from one type of fodder beet about 200 years ago. However, wild beet (B. vulgaris subsp. maritima) has a very wide germplasm base but has been little exploited. The major constraint to economical sugarbeet production is plant disease and other disorders, in part due to its narrow germplasm base and vulnerability to disease. In each region of the USA, different diseases have become important. In the western USA, these include rhizomania, caused by BNYVV, virus yellows caused by a complex of aphid-vectored closteroviruses and luteoviruses, curly top, caused by beet curly top virus, powdery mildew, caused by Erysiphe polygoni, cercospora leaf spot, caused by Cercospora beticola, nematodes and a number of soil-borne fungi and bacteria that cause damping-off, seedling loss, and root rots. Environmental stress such as exposure to cold temperatures that lead to detrimental early bolting, flowering, and seed production can be problems as are sustained high temperatures. To remain economically competitive, sugarbeet must also continue to be improved for agronomic, yield, and quality factors. The best, sometimes only, and most environmentally safe means to alleviate these production problems and hazards are with genetic host-plant resistance and improved performance. To solve these problems, a comprehensive research program on germplasm enhancement, breeding, and genetics is being carried out. This includes searching for useful host-plant resistance genes within cultivated sugarbeet, other cultigens of beet, and its wild, ancestral species, particularly B. vulgaris subsp. maritima. A long-term population improvement program is being used to enhance advanced sugarbeet breeding lines with new resistance genes and to improve agronomic, yield, and quality characteristics. Plant disease and low productivity have the potential to eliminate the beet sugar industry from various regions of the USA. Chemical control for some of the hazards of production has been or is being lost or is very expensive. Chemical and cultural controls for other problems do not exist. Host-plant resistance and improved production may be the only long term, environmentally friendly way to economically produce sugarbeet. Where sugarbeet is grown efficiently, it provides growers with a sound and productive rotational crop and provides a large number of jobs to the rural community through harvest, processing and refining payrolls. Through their multiplying effect, sugarbeet production and processing provide the whole rural community with significant economic and social benefits that are not easily replaced by other crops. 2.List the milestones (indicators of progress) from your Project Plan. Germplasm enhancement: Yr. 1-5 - Run a comprehensive population improvement project for sugarbeet. Each year monogerm and multigerm populations will be cycled. Monogerm populations and lines indexed for O-type and improved monogermity. Backcrosses to CMS's. Progeny families generated. Breeding lines, source populations, progeny families, synthetics, and experimental hybrids evaluated for individual and combined disease resistance and components of sugar yield and genetic parameters. Replicated variety trials run at Salinas and Brawley and appropriate disease inoculations made or promoted. Data from trials analyzed and summarized reports to cooperators in annual reports and presentations. Accessions from PI Station evaluated for disease and agronomic traits. Data entered into GRIN. PI lines or individual plants that show promise selected and introgressed in sugarbeet germplasm. Prebred and enhanced populations searched for useful genetic variability. Germplasm lines released yearly or as new, improved, or unique combinations are developed. Released germplasm to NSSL and PI Station's working collection and published in CROP SCIENCE as Registrations. Resistance to Rhizoctonia and Cercospora: Yr. 1-5 - In alternating cycles at Salinas and Ft. Collins, existing and new source populations generated and advanced to combine resistance to Rhizoctonia/Cercospora with rhizomania/curly top/bolting resistance. Each year, one phase of each population in every other cycle selected and evaluated at Salinas for resistance to rhizomania, bolting, curly top, etc., and components of sugar yield. Yr. 4 or 5 - Germplasm released and registered with Panella. Molecular markers Powdery Mildew: Yr.1 - Marker analyses finished. Yr. 2 - Publication with Weiland. Yr. 3&4 - Confirm linkages and efficiency of MAS. Yr. 5 - Publication. Curly Top QTL: Yr. 2-4 - With progenies and lines from McGrath, phenotype for resistance reaction at Kimberly, ID and in Salinas greenhouses using seedling reactions and ELISA data. Yr. 5 - Analyze data and manuscript with McGrath & Panella. Beet mosaic: Yr. 1 - From conventional genetic analysis, publish linkage between Bm and A. From ongoing research, confirm molecular genetic linkages (Weiland). Yr. 2 - Publication. Erwinia: Yr. 1 - From existing F2's, create divergent bulks and F3 (S2) lines. Yr. 2-3 - Identify linked markers and confirm efficacy of MAS. Yr. 4 - Publish. Resistance to rhizomania: Yr. 1-5 - Continue ongoing selection and evaluation program. Search germplasm resources for new reaction genes. Under high temperatures: Yr. 1-2 - From population C51 (sugarbeet x B.v. maritima), S2 derived S3 progenies evaluated and scored at Brawley for survival at high temperatures under severe rhizomania. Yr. 3 - Elite lines increased and retested. Genetic analyses. Yr. 4 - Publication if interpretations possible. BNYVV systemic infection: Yr. 1 - With Virology CRIS at Salinas, preliminary F2 data presented and published in Proceedings. S2 and S3 progenies generated and evaluated by mechanical inoculation. Yr. 3 - Published and unique lines increased. Reaction of other viruses tested. Races/Strains of BNYVV: Yr. 1 - With Virology CRIS at Salinas, run and analyze soils (virus strains) x varieties (genotypes Rz vs rzrz) for BNYVV titer interactions. Yr. 2 - If warranted, screen known existing sources of resistance to new race (strain) in greenhouse tests. Inoculate and establish infested soil test areas at Brawley/Salinas. Publish existence of new race and resistance reactions. Yr. 3-5 - Initiate new breeding program for race 2 of BNYVV. Beet soil-borne mosaic virus: Yr. 1 - Publish completed interaction tests of varieties (Rz vs rzrz) x soils (healthy, BNYVV, SBMV, P. betae vector only, and combinations) with Virology CRIS. Yr. 1-2 - With Virology CRIS, initiate greenhouse and microplot screens for varietal reaction (ELISA value) to BSBMV. Yr. 3 - If resistance or genetic variability identified, publish. Beet oak-leaf virus: Yr. 1 - With Virology CRIS, initiate exploratory greenhouse screens for varietal reaction, sources of resistance, and economic damage. Resistance to bolting and other diseases Bolting: Yr. 1-5 - Emphasis on improving nonbolting tendency for overwintered productions. From existing program, release and registrar high nonbolting lines. Beet chlorosis virus: Yr. 1 - Based upon 2 existing years of data, run field tests to confirm differential reaction to BChV in selected lines. Yr. 2 - Present and publish. Virus yellows resistance: Yr. 1-5 - Continue selection for resistance to virus yellows complex. Search wild species for additional resistance. Powdery mildew: Yr. 1-5 - Check research plots of Pm entries for loss of resistance. Determine if Pm from WB97 or WB242 reacts differently. If observed and confirmed, publish existence of new race(s). Continue resistance breeding for slow-mildew and search for additional major reaction genes. Resistance to nematodes Root knot nematode: Yr. 1 - Develop a molecular genetic marker(s) of root-knot nematode resistance for non'Mi-1' series of sugarbeet, release one nematode-resistant sugarbeet germplasm, publish papers on (a) an analysis of Meloidogyne spp. resistance related isozyme markers, and/or (b) inheritance of root-knot nematode resistance in sugarbeet. Yr. 2 - Release one improved nematode-resistant sugarbeet germplasm, and register one sugarbeet germplasm in CROP SCIENCE, induce leaf-explants and ovules in vitro to regenerate and/or multiply elite sugarbeet genotypes, and conduct resistant sugarbeet field trials at different locations and conditions. Yr. 3 - Start combining the two known sources (alleles or genes) of Meloidogyne spp. resistance (Mi-1 and M66) into one single sugarbeet genome, examine mitotic and meiotic chromosomal stability of selected breeding populations, release improved sugarbeet breeding lines to be useable as pollen parents of commercial hybrids. Yr. 4 - Continue breeding research on sugarbeet with multiple sources of Meloidogyne spp. resistance, complete MAS protocols for the known resistance types, conduct extensive field trials for the advanced nematode resistance breeding materials. Yr. 5 - Complete development of sugarbeet with multi-source Meloidogyne spp. resistance against M. incognita, M. javanica, M. arenaria, M. hapla, M. chitwoodi, and M. fallax, release elite sugarbeet germplasms with high level of nematode resistance, agronomic characteristics, and combining ability to the beet sugar industry, and publish articles on the development of nematode resistant sugarbeet. Cyst nematode: Yr. 2-5 - Research on nematode resistance will be redirected to sugarbeet cyst nematode resistance. Beta germplasm resources will be screened to search for new, useful resistance. Greenhouse and field tests to measure efficacy of resistance will be run. If resistance is found, germplasm developments will be released. 3.Milestones: Germplasm enhancement - All phases were completed as scheduled and germplasm lines were released and registered. Milestones substantially met. Rhizoctonia & Cercospora resistance - Germplasm lines FC201 and FC301 were released. New populations potentially combining listed resistance were selected for resistance to rhizomania. Milestones met. Molecular markers - Marker work for powdery mildew was completed and manuscript is in preparation. Curly top progenies were generated. Beet mosaic manuscript is being prepared. Erwinia progenies were phenotyped. Milestones were substantially met. Resistance to Rhizomania - Ongoing selection and evaluation program goals were fully met. Progenies from C51 were developed and scored under severe conditions in Imperial Valley. Data to date were published in IIRB-ASSBT Proceedings. Soils and genotypes were evaluated against BNYVV for the occurrence of races of BNYVV and new reaction genes. Research on BSBMV and BOLV was delayed because of other important program considerations. Resistance to bolting and other diseases - Progeny families were evaluated in overwintered plantings for nonbolting tendency. Field tests to evaluate differential reactions to BChV were completed. Tests to specifically evaluate reaction to BYV were initiated. Lines and progeny families with powdery mildew resistance from WB97 andWB242 were evaluated in field trials. Milestones were substantially met. Resistance to nematodes - Research on resistance to RKN was completed a molecular marker for resistance allele R6m-1 was published. Germplasm line M6-2 was released and registered. Milestones were completely met. Research and resistance breeding on cyst nematode were initiated. Germplasm enhancement - 2005, 2006, 2007: Population improvement and a comprehensive breeding and research program will be continued. New sources of resistance and improvements will be released as prudent. Releases will be registered. Releases and new sources of resistance will continue to be critical to the continued increased productivity of sugarbeet. Rhizoctonia & Cercospora resistance - 2005, 2006, 2007: Selections for resistance to rhizomania, bolting, and other disease resistance as well as for improvements in productivity will be made. The combination of resistance for rhizomania with resistance to rhizoctonia and/or cercospora leafspot will add substantially to the usefulness of this germplasm for the sugarbeet industry. Molecular markers - 2005, 2006, 2007: The identification of molecular markers for resistance to powdery mildew will be published. Pm from the two different sources will be tested for allelism. Seedings screened by the identified markers will be biologically evaluated to confirm their usefulness. In 2005, phenotyping for reaction to curly top will be started. In 2006, seed from divergent selections will be produced and in 2007, re-phenotyped in inoculated field tests. In 2005, the occurrence of a molecular marker for BMV resistance will be published. In 2005 and 2006, progenies will be generated and phenotyped for reaction to Erwinia. Resistance to rhizomania - 2005, 2006, 2007: The basic breeding and research program to develop and evaluate resistance to rhizomania will be continued. In 2005-2007, progenies will be evaluated under high temperatures in Imperial Valley to identify genotypes with high resistance. Research on systemic infection of BNYVV from local lesions will be relaxed because of the need to address the emergence of new pathotypes of the virus that defeat the most widely used major gene Rz1. In 2005, new nurseries will be established with the new race of BNYVV. In 2006 and 2007, all available beet genetic resources will be screened and evaluated for reaction to the new emerging races. If possible, new sources of resistance will be identified and selected, genetic analyses made, and isolate x genotype interactions determined. Because of the seriousness of the new emerging races of BNYVV, research on BSBMV and BOLV will be delayed. Without the identification and deployment of new resistance genes to rhizomania (BNYVV), the new resistance genes to rhizomania (BNYVV), the sugarbeet industry could be in jeopardy. Resistance to bolting and other diseases - 2005, 2006, 2007: All generated breeding materials will be evaluated for bolting tendency each year. Those with too high levels will be discarded. In 2005, a manuscript on the reaction of sugarbeet to BChV will be written and submitted for publication. In 2006, it will be published. In 2005-2006, inoculated field tests will be used to determine the reaction of sugarbeet breeding lines to BYV. In 2007, a manuscript will be written and submitted for publication. In all years, Pm lines will be grown in the field to evaluate for the emergence of new race(s). Broadly based sugarbeet x wild beet populations will continue to be screened for new resistance genes. Resistance to nematodes - 2005-2007: The research and breeding program on RKN will be phased out. This is due to a change in research personnel that this program has been highly successful to date providing two new resistance sources and markers for these resistance genes, and nationally and internationally, sugarbeet cyst nematode (SBCN) is much more important. In 2005-2007, specific germplasm will be evaluated for reaction to SBCN. From the rhizomania resistance breeding project, it has been observed that there appears to be segregation for resistance or tolerance to SBCN. In 2005, germplasm will be evaluated on a single plant basis in greenhouse tests. Field tests will be used to evaluate the efficacy of this resistance. In 2006, screening and selection will be continued and divergent selections will be made. In 2007, research on molecular genetic markers will be done and improved SBCN resistant lines will be released. Field tests on the strength and agronomic value of this resistance will be published from tests grown in 2004-2006. If obtaining resistance to SBCN from the wild beet germplasm is successful, it will materially add to the productivity of sugarbeet worldwide and alleviate the total dependence upon rotations, soil fumigants, and pesticides. 4.What were the most significant accomplishments this past year? A. Single Most Significant Accomplishment during FY 2004 year: Root knot nematodes (Meloidogyne spp.) can cause significant damage in sugarbeet in subtropical environments and no host-plant resistance was previously known in sugarbeet and protection against these pests required the use of soil fumigant, nematicides, or consideration of crop rotation and nematode prevalence. The research was lead by M.H. Yu of the U.S. Agricultural Research Station, Salinas, Calif., in collaboration with Dr. R.T. Lewellen at the Salinas Station, and J.J. Weiland at the USDA-ARS in Fargo, ND. Germplasm line M6-2 was released and registered joining earlier releases from this project to give sugarbeet breeders sources of high resistance to root knot nematode and sources of resistance that may be useful in the future in molecular genetic research as a resistance gene that gives protection across a wide spectrum of root knot nematode species. The project at Salinas searched the wild species of beet and identified two different sources of resistance, transferred these resistances into sugarbeet, showed that they conditioned high resistance to at least six separate species of Meloidogyne, identified an isozyme marker for one of the resistance genes, identified a molecular marker for the resistance allele of M6-2 and showed that it was inherited as a single dominant allele named R6m-1. B. Other significant accomplishment(s), if any: None C. Significant Activities that Support Special Target Populations: None D. None 5.Describe the major accomplishments over the life of the project, including their predicted or actual impact. Sugarbeet germplasm enhancement over a broad area and for individual and combined sources of resistance is the major accomplishment of this project and includes research bridging from previous projects and is documented by the releases and registrations of germplasm lines (see Scientific Publications in this report). Collectively, these germplasm developments provide resistance to rhizomania (Beet necrotic yellow vein virus), virus yellows (Beet chlorosis virus and Beet yellows virus), Curly top virus, bolting, Erwinia, powdery mildew (Erysiphe polygoni), root knot nematodes (Meloidogyne spp.), and cyst nematode (Heterodera schachtii). Resistances include qualitatively inherited sources identified in wild species and transferred to sugarbeet for rhizomania, powdery mildew, root knot nematode, and cyst nematode. Resistances also include quantitatively inherited sources against curly top, virus yellows, bolting, powdery mildew and others. In addition to their other attributes, it appears that C927-4, CP04, CP07, and CP08, as well as other yet unreleased germplasm lines, may have moderately high resistance or tolerance to sugarbeet cyst nematode from wild beet line-242 as well as others. This is particularly noteworthy because of the very extended history of cyst nematode resistance breeding without achieving commercial success. Based upon history and the use of previous releases from this project, it can be anticipated that these releases will be important in the development of commercial cultivars and the amelioration and control of disease and pest problems in sugarbeet and the overall improvement in productivity. 6.What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end-user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? Lewellen, R.T. Development of sugarbeet breeding lines and germplasm. Sugarbeet Research, 2003 Report, 2004. p. A36-A175. (Report on research program to the Beet Sugar Development Foundation. Distributed widely by BSDF to members and sugarbeet research community). Lewellen, R.T. Breeding sugarbeet for combined resistance to diseases. Presented at 2004 Sugarbeet Research Review, Holtville, CA, January 14, 2004. Lewellen, R.T. 2003. Field day for the California sugarbeet industry at Salinas and review of funded research projects, Salinas, CA, September 9, 2003. Lewellen, R.T. 2003. Discussion, field day and tour of sugarbeet research for Seedex and Desprez breeders, Salinas, CA, September 11-12, 2003. Lewellen, R.T. 2004. Seminar on Sugarbeet Population Improvement, U.S. Agric. Res. Stn., Salinas, CA, May 22, 2004. Lewellen, R.T. 2003. Rhizomania strains and resistance. Rhizomania Conference, Alexandria, MN, August 27, 2003. (Presentation to the sugarbeet industry of the upper mid-west.) Release of sugarbeet germplasm lines by R.T. Lewellen in 2003-2004: Notice of release of combined powdery mildew and rhizomania resistant sugarbeet germplasm lines CP03, CP04, CP05, and CP06. Notice of release of combined powdery mildew and rhizomania resistant sugarbeet germplasm lines CP07 and CP08. Notice of release of high performing sugarbeet germplasm line C81-22. Notice of release of sugarbeet germplasm lines C842 and C842CMS with resistance to curly top and rhizomania. Release of sugarbeet germplasm lines by L.W. Panella and R.T. Lewellen in 2004: Notice of release of FC201 monogerm, O-type sugarbeet germplasm with resistance to rhizomania and other diseases. Notice of release of FC301 monogerm, O-type sugarbeet germplasm with resistance to rhizomania and cercospora leafspot. 7.List your most important publications in the popular press and presentations to organizations and articles written about your work. Yu, M.H. 2004. Breeding sugarbeet that fends off nematodes. The California Sugar Beet 2003 Annual Report. p. 10-11. Liu, H.-Y., Sears, J.L., and Lewellen, R.T. 2004. P-pathotype of rhizomania in sugar beet has not been identified in the Imperial Valley. The California Sugar Beet 2003 Annual Report. p. 12-13. Review Publications Yu, M.H., Lewellen, R.T. Registration of root-knot nematode-resistant sugarbeet germplasm M6-2. Crop Science. 2004. v. 44. p.1502-1503. Lewellen, R.T. Registration of rhizomania resistant, monogerm populations c869 and c869cms sugarbeet. Crop Science. 2004. v.44. p.357-358. Lewellen, R.T. 2004. Registration of sugarbeet germplasm lines c67/2, c69/2, c78/3, and c80/2 with resistance to virus yellows and rhizomania. Crop Science. 2004. v. 44. p. 358-359. Lewellen, R.T. Registration of sugarbeet germplasm lines c927-4, c929-62, c930-19, and c930-35 with resistance to rhizomania, virus yellows, and bolting.. Crop Science. 2004. v. 44. p. 359-361. Wisler, G.C., Lewellen, R.T., Sears, J.L., Wasson, J.W., Liu, H., Wintermantel, W.M. Interaction between beet necrotic yellow vein virus and beet soil-borne mosaic virus in sugar beet. Plant Disease. 2003. v. 87. p. 1170-1175. WEILAND, J.J., YU, M.H. A CLEAVED AMPLIFIED POLYMORPHIC SEQUENCE (CAPS) MARKER ASSOCIATED WITH ROOT-KNOT NEMATODE IN SUGAR BEET. CROP SCIENCE. 2003. v. 43. p. 1814-1818. McGrath, J.M., Lewellen, R.T. 2004. Registration of EL0204 sugarbeet germplasm with smooth-root and resistance to rhizomania. Crop Science. 44(3):1032-1033. YU, M.H. DEVELOPING SUGARBEET WITH RESISTANCE TO MELOIDOGYNE SPP. INTERNATIONAL CONGRESS OF GENETICS PROCEEDINGS. 2003. p. 163. YU, M.H. DEVELOPMENT OF ROOT-KNOT NEMATODE-RESISTANT SUGARBEET. INTERNATIONAL INSTITUTE FOR BEET RESEARCH PROCEEDINGS. 2003. p. 763-765.