Progress 09/01/13 to 08/31/17
Outputs
Target Audience:The main target audiences reached over the course of this grant include: Research directors and tomato breeders in the private sector (seed company directors of research and tomato breeders working for seed companies). This group is crucial for the insect resistance trait to be incorporated into new commercial tomato cultivars Tomato breeders in the public sector (university and USDA scientists working on tomato genetics or tomato breeding, who will be working using the trait in future lines adapted to diverse growing conditions nationally Public sector scientists and extension specialists focused on control of insects and insect transmitted virus in tomato. These are either entomologists or plant pathologists. Interesting this group in the materials is important to start trials needed to determine the optimal means to use this trait within a balanced IPM program. Research scientists in both the private and public sector in allied fields (ecology, chemical ecology, molecular biology of secondary metabolites). These scientists have become very interested in the acylsugar producing tomato lines as materials to use in study of insect/plant interactions, insect/predator or parasite interactions, and in the biosynthesis of bioactive secondary metabolites, and the evolution of these pathways. Undergraduate and graduate students who toured the research fields, which were used to teach tomato breeding and integration of disciplines (in this case, genetics, plant pathology, entomology) to solve complex problems. The ultimate target audiences will be tomato growers, when the acylsugar mediated resistance can be used in commercial hybrids for use in tomato production. Changes/Problems:As described in the "accomplishments" section, a change in the means for distributing information was changed to be able to provide more detailed information in a more useful form. What opportunities for training and professional development has the project provided?Opportunities for Training and Professional Development An undergraduate student intern was mentored in a research project using the acylsugar lines during the fall 2014/spring 2015 research program. The purpose of the internship was both to provide the undergraduate student with a hands-on research experience, and to train the graduate student acting as the mentor in how to be an effective mentor. The intern presented her results in a cyber symposium in May 2015. The program was overviewed and plants shown to first year Cornell graduate students and senior undergraduates in a lab course each fall in 2014, 2015, 2016, and 2017. The graduate student developing the new acylsugar lines for this project presented his results in posters or as a speaker in multiple venues. These opportunities provided experience in development and presentation of posters and seminars, and gave him opportunities to present his work to and interact with professionals in the discipline during his graduate training. The presentations include those listed below. A. POSTER PRESENTATIONS: The National Association of Plant Breeders meeting Summer of 2014: "Utilizing Acylsugar Chemistry to Prebreed Tomatoes for Optimal Pest Control" National Association of Plant Breeders Fall 2017: "Prebreeding Tomato For Optimized Acylsugar-Mediated Resistance To Insects" B. PRESENTATIONS AS SPEAKER: The international meeting of the Tomato Breeders Roundtable Fall of 2014: "Utilizing Acylsugar Chemistry to Optimize Tomato Pest Control" The 10th International Symposium on Thysanoptera and Tospoviruses, May 2015: "Utilizing Acylsugar Chemistry for Optimal Thrips and Tospovirus Control in Tomato" National Association of Plant Breeders Presentation 2017: "Modifications of Acylsugar Chemistry to Optimize Insect and Virus Resistance in Tomato" The graduate student was a speaker at the School of Integrative Plant Science Cornell Symposium, Fall 2016: "Breeding to Replace Pesticides with Biodeterrence: Modification of Acylsugar Chemistry to Optimize Natural Pest Control" C. FIELD DAY PRESENTATIONS The Vegetable Breeding Institute at Cornell annually holds 2 day field days attended by 35 to 45 representatives of national and international vegetable seed companies. During these field days, the progress on current vegetable breeding programs are displayed through discussions in the fields. The graduate student on this project presented his work yearly at these field days in 2013, 2014, 2015, and 2016. This gave him experience in field based presentations. The graduate student also had opportunities to meet with seed company representatives in group and individual settings, which helps with networking and professional development. How have the results been disseminated to communities of interest?Dissemination of results For year 3: 9/1/16 to 8/31/17 A Mini-seminar was presented by the graduate student (Smeda): in the School of Integrative Plant Science Cornell Symposium, Fall 2016: "Breeding to Replace Pesticides with Biodeterrence: Modification of Acylsugar Chemistry to Optimize Natural Pest Control" A poster was presented by Mutschler and Smeda: at the National Association of Plant Breeders, Davis CA, Aug 2017: "Prebreeding Tomato For Optimized Acylsugar-Mediated Resistance To Insects" A seminar was presented by Smeda: at the National Association of Plant Breeders Presentation, Davis CA, Aug 2017: "Modifications of Acylsugar Chemistry to Optimize Insect and Virus Resistance in Tomato" Vegetable Breeding Institute field day, Aug 28, 29 2017: The results of the project were also presented to 40 seed company representatives at the annual field days for the Vegetable Breeding Institute throughout the period of this grant. Handouts used for the field demonstrations included acknowledgment of source of funding. PUBLICATIONS: the three papers already published are listed on the products page. Three additional publications are being written and will be submitted spring of 2018 IMPACT OF PRESENTATIONS: The presentations routinely raised great interest in the work and its potential to control insect pests and viral diseases of tomato. It also resulted in offers of assistance to expand testing materials in development at numerous sites in the US and abroad, and indications of interest in use of the materials in development for research in control of a wide range of current and emerging insect pests, as well as virus control in tomato. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
GOAL 1. Transfer QTL altering acylsugar chemotype to acylsugar tomato line CU071026, producing sets of sister-lines plus and minus different QTL, and characterize these lines for acylsugar chemotype and level. Over the course of this project the acylsugar lines with 1 to 3 QTL altering fatty acids were created: FA2/AS, FA5/AS, FA7/AS, FA8/AS, FA2/FA7/AS, FA2/FA8/AS, FA7/FA8/AS, and FA2/FA7/FA8/AS, as well as the acylglucose producing line AG3/AG4/AG11/AS. The nine related acylsugar lines and appropriate controls were grwn simultaneously grown and characterized for acylsugar level (using the acylsugar assay), for acylsugar fatty acid content (using gas chromatography mass spectrometry analysis of derivatized acylsugars), and for total profile of accumulated acylsugars (using liquid chromatography mass spectrometry). Sophisticated interactions between the acylsugar chemotype QTL were observed that additively and epistaticly impacted acylsugar levels and chemistry. The QTL in FA5/AS almost completely shut off acylsugar production due to a missing epistatic QTL. Similarly, the QTL in FA8/AS governed a major shift in acylsugar fatty acid profile, but simultaneously reduced acylsugar level. The combination of the QTL in FA2/AS and FA7/AS resulted in a significant increase in acylsugar level due to epistatic interactions. The complete information on the generation of the nine acylsugar lines, and their characterizations, is included in three journal, one published in 2016 and two published in 2017. The knowledge gained on how the introgressed QTL impact acylsugar chemotype, will greatly informs utility of these QTL in further breeding efforts to generate elite tomato lines with optimal acylsugar-mediated resistance. GOAL 2. Determine the impacts of different acylsugar chemotypes produced by the sister lines against whiteflies (FL) and thrips (NC). Trials using the new acylsugar lines against whiteflies (FL) and thrips (NC) were performed. FLORIDA (co-PI Smith): Performed field trials in the spring and fall 2014 and spring 2015 at the U-FL Gulf Coast Research and Education Center to evaluate efficacy of the new acylsugar producing tomato lines in suppressing sweetpotato whitefly (Bemisia tabaci) and incidence of Tomato yellow leaf curl virus (TYLCV). Data were collected on densities of whitefly eggs, first, mid and fourth instar nymphs. Foliage samples were collected for analysis of acylsugar fatty acid composition. Foliage was also collected to test for the presence of TYLCV using PCR. The data revealed that the new tomato lines with higher acylsugar levels exhibited greater resistance to whitefly oviposition and nymph survival. In addition, some lines producing different types of acylsugars had 0% of plants being TYLCV infected, compared to ca. 20% of CU070126 plants and 35% of tomato controls being TYLCV infected. Furthermore, acylsugar lines and hybrids producing unaltered acylsugars, similar to those of CU071026, but which were homozygous or heterozygous (respectively) for the TYLCV resistance gene Ty-3, also showed 0% TYLCV infection even though 35% of a tomato hybrid heterozygous for Ty-3 were TYLCV infected. This indicates that the combination of production of the most effective level/types of acylsugars with virus resistance genes could be a successful strategy for control of insect vectored virus. A manuscript for publication of this data is currently being drafted. NORTH CAROLINA (co-PI Kennedy): Conducted field trials on a commercial tomato farm to evaluate efficacy of tomato lines from the Cornell breeding program in suppressing tobacco thrips and western flower thrips populations and incidence of the thrips-transmitted virus, Tomato spotted wilt orthotospovirus (TSWV). In the summer of 2014 we evaluated 14 tomato entries and in the summer of 2015 we evaluated 15 entries. Thrips samples were collected for identification to species and identifications were performed. Foliage samples were collected for analysis of acylsugar composition and foliage was also collected and tested for TSWV infection using ELISA. Data was summarized and analyzed for thrips numbers and TSWV incidence on each entry evaluated. The thrips and TSWV data revealed that entries with higher levels of acylsugars had fewer thrips on the plants and demonstrated a reduction in TSWV infection. A publication summarizing the field trials in NC is currently being drafted. The growth of the acylsugar-producing tomato lines for insect trials in different states with very different climates also provided an opportunity to determine impact of environment on acylsugar level and types. Characterization of these lines for various aspects of acylsugar chemistry demonstrated such high heritability of acylsugar chemistry under different conditions. However level of acylsugars was found to differ quite significantly under different environmental conditions. The breeding implications of this fact are that different acysugar lines might be needed in different locations to ensure that sufficient acysugar levels are produced by tomato hybrid used in each location, but that only minimal screening for acylsugar chemistry would be necessary during transfer of the underlying QTL/genes in breeding of elite germplasm. GOAL 3. Use genotypic data generated to develop/test acylsugar breeding SNP sets for accelerated introgression of QTL for acylsugar production and controlling acylsugar chemotype into tomato cultivars. The lines generated during this project have been genotyped using a variety of methods, including use of CAPS markers, SNP panels, and genotyping by sequencing (GBS). We have sufficient markers to create a master list that can be used for selection and rapid transfer of select introgressions, and also for reduction of introgression size through selection of recombinant plants. The major outcome of this objective was knowledge about the size and specific breakpoints for all the acylsugar chemistry QTL introgressions. This knowledge has already been utilized and disseminated through three the manuscripts published during this project timeline; further use of the marker information gathered during this project will greatly facilitate the rapid breeding and selection of elite tomato lines and hybrids with reduced acylsugar chemotype QTL introgression sizes. As discussed in Goal 4, we are preparing to upload further information on markers and genomic characterizations on a web site for use by the growing community of breeders and researchers interested in utilizing acylsugar mediated insect resistance. GOAL 4. Develop content (webinars) describing this project, tomato lines available, and teaching methods to breed tomatoes with optimal acylsugar chemotypes for dissemination to breeders via PBGCoP eXtension). We the work in objectives 1 to 3 rapidly progressed, it became obvious that this goal should be modified. The information that is most useful and necessary for researchers and breeders is the detailed methodologies needed to work with this trait, such as the acylsugar assay, as well as detailed characterization data from GC and LCMS analyses and the methods used to obtain this data. None of this type of information is well suited to webinar presentation. So we generated detailed information about the characterization of the germplasm from this project, both for aspects of acylsugar chemistry, as well as genotypic information. This information has been published over three main publications in Molecular Breeding (2016, 2017 and 2017), with extensive additions in supplemental materials included in these publications. A spreadsheet with full information on PCR based markers (marker genomic location, primer sequence, amplification, restriction enzymes and conditions, and band sizes) to be used for each S. pennellii introgression. We are editing these protocol for uploading, in January or February of 2018, to a publically accessible website.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Smeda JR, Schilmiller AL, Kessler A, Mutschler MA. 2017. Combination of QTL affecting acylsugar chemistry reveals additive and epistatic genetic interactions to increase acylsugar profile diversity. Molecular Breeding. 37(8):104
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Smeda JR, Schilmiller AL, Anderson T, Ben-Mahmoud S, Ullman DE, Chappell TM, Kessler A, Mutschler MA. 2017. Combination of Acylglucose QTL reveals additive and epistatic genetic interactions and impacts insect oviposition and virus infection. Molecular Breeding. doi:10.1007/s11032-017-0756-z
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Progress 09/01/15 to 08/31/16
Outputs
Target Audience:The main target audiences reached during this period include tomato breeders in both the private sector (seed company directors of research and tomato breeders working for seed companies) and in the public sector (university and USDA scientists working on tomato genetics or tomato breeding, or on entomology/plant pathology of tomato). An additional target audience were undergraduate students who toured the research field, which were used to teach tomato breeding and integration of disciplines (in this case, genetics, plant pathology, entomology) to solve complex problems. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The graduate student developing the new acylsugar lines for this project (at Cornell in NYS) presented his results at the the Aug 2016 VBI field day to over 40 international seed company representatives. The presentation was very well received. This graduate student was the sole invited graduate student presenter for the Section of Plant Breeding at the Oct 2016 School of Integrated Plant Science meeting Nov 2016 at Cornell. How have the results been disseminated to communities of interest?The results of the project were also presented to 40 seed company representatives at the annual field day for the Vegetable Breeding Institute, Aug 2016, Ithaca NY. (Slide presentation included acknowledgment of source of funding). This presentation resulted great interest among seed company representatives, who have requested seed of lines. A paper detailing the transfer of single fatty acid QTL into CU071026 to produce the lines FA2/AS, FA7/AS, FA5/AS, and FA8/AS, fine mapping the QTL to regions as short as 500,000 bp, and fully characterizing the acylsugars produced by each of these lines using the acylsugar assay, GC, and LCMS was accepted for publication by Molecular Breeding. It is in press, and we expect publication during Dec 2016 or Jan 2017. Acknowledgment of source of funding was included in this paper A paper detailing the combination of two or three fatty acid QTL into CU071026 to produce the lines FA2/ FA7/AS, FA2/FA8/AS, FA7/FA8/AS and FA2/ FA7/FA8/AS, and fully characterizing the acylsugars produced by each of these lines using the acylsugar assay, GC, and LCMS has been prepared and is in pre-submission review. We expect to submit the manuscript during Dec 2016, for publication in 2017. Acknowledgment of source of funding will be included in this paper What do you plan to do during the next reporting period to accomplish the goals?Goal 1. Increase seed supplies of lines; as more researchers want to use the new acylsugar lines, we must have sufficient seed supplies. It would be of interest to transfer the fatty acid QTL into the acylglucose line. Crosses have been made to start the process, that will facilitate the transfer, but completion of that work would be beyond the term of this grant. The acylsugar sister lines all share the poor fruit set/seed set and small fruit size of the acylsugar benchmark line CU071026. We need to improve fruit set/seed set/fruit size of lines. A separate project has identified regions of QTL that appear to interact to control these traits, and initial crosses are being start the modifications needed to improve fruit set/seed set in the lines possessing fatty acid or acylglucose QTL, however that process will probably not be completed in the term of this grant due to time constrains/time required per generation in tomato. Goal 2. Work is largely completed on field trials of the impact of the acylsugar lines on thrips (NC) and whiteflies (FL). Work to be completed in the last year of the grant are full data analyses of these data sets and preparation/submission of resulting publications to appropriate journals in 2017. Work is also completed on laboratory tests of the impact of the acylsugar lines on larval growth and survival of Spodoptera exigua (NY). Analysis of the resulting data analysis is largely completed; a manuscript will be submitted for publication, hopefully in early 2017. The acylsugar sister lines have also been shared with cooperators who are not co-PIs on this grant, to provide additional testing of the impact of the lines on thrips, whiteflies, and the transmission of virus by these insect vectors. During 2017 we will also be preparing publications on the results of those laboratory studies. Goal 3. We are creating a master report of markers that can be used for the transfer and reduction of each of the introgressions in the benchmark line CU071026 or the sister lines with modified acylsugar chemotypes. This includes a large set of PCR based markers (CAPS, or markers derived from SNPS) that fall within the introgressions associated with each of the fatty acid or acylglucose QTL, as well as the introgressions characteristic of the benchmark line CU071026 and all of the derived acylsugar sister lines. In addition, we have identified many SNPS within each of these introgressions that could be to provide additional markers.As the QTL have been mapped within each introgressions, the markers that are closest to the QTL are identified. Goal 4. We are rethinking this goal. It is premature to present conclusion on the best acylsugar lines for broad insect control, as we have only tested three insect species. More appropriate and useful information to provide at this time are detailed instructions on the methods used for the acylsugar colorimetric assay for acylsugar level, and the parameters of GC and LCMS characterization. However due to the details involved in the protocols, these are not well suited to webinar format. So we are considering other options to provide meaningful information to the research and breeding communities.
Impacts
What was accomplished under these goals?
GOAL 1. Building on the work of years 1 and 2, in year 3 the single QTL acylsugar lines FA2/AS, FA7/AS and FA8/AS, and the multiple QTL lines FA2/FA7/AS, FA2/FA8/AS and FA2/FA7/FA8/AS, as well as controls and the acylglucose producing line AG3/AG4/AG11, were grown simultaneously, characterized for acylsugar level using the acylsugar assay, for fatty acid content using GC analysis of derivatized acylsugars. In addition, samples from all entries were also analyzed for acylsugar structures using LCMS, which is the first time this technique was used for the lines in development. This combined analysis provided more detailed characterizations of the acylsugars produced by each line under common conditions. The results also confirmed prior observation that the FA8/AS line produces lower acylsugar levels than the other acylsugar lines. One unexpected outcome was that the switch from production of acylsucroses to acylglucoses in the AG3/AG4/AG11 acylglucose line is accompanied by a modest switch in fatty acid profile. GOAL 2. Work started in year 2 was substantially expanded in year 3 testing these lines for impact on growth/survival of Spodoptera exigua larvae fed on leaves from these lines, on thrips oviposition and inoculation of the tospovirus TSWV by viliferous thrips, and on Bemisia whitefly oviposition and inoculation of the begomovirus TYLCV by viliferous whiteflies. The work has progressed sufficiently that we are now preparing at least three papers for submission in 2017 GOAL 3. Some of lines were tested using SNP panels and more were tested using genotyping by sequencing. We now have sufficient markers to create a master list of markers that can be used for selection and rapid transfer of select introgressions, and also for reduction of introgression size through selection of recombinant plants. GOAL 4. We progressed to the point that it became obvious that this goal should be modified. The most useful information to provide researchers and breeders is detailed methodologies, such as the acylsugar assay and master marker files, as well as detailed characterization data from GC and LCMS analyses. None of this type of information is well suited to webinars. So we are developing the most useful information in the most useful formats and consider the best way to provide ready access to the information.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Leckie, BM, D'Ambrosio, DA, Chappell, TM, Halitschke, R, De Jong, DM, Kessler, A, Kennedy, GG, Mutschler, MA 2016. Differential and synergistic functionality of acylsugars in suppressing oviposition by insect herbivores. PLOS ONE DOI:10.1371/journal.pone.0153345
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Smeda John R., Anthony L. Schilmiller, Robert L. Last, Martha A. Mutschler. 2016 Introgression of acylsugar chemistry QTL modifies the composition and structure of acylsugars produced by high-accumulating tomato lines" Molecular Breeding. DOI 10.1007/s11032-016-0584-6
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Progress 09/01/14 to 08/31/15
Outputs
Target Audience:A major target audience reached during this period include tomato breeders in both the private sector (seed company directors of research and tomato breeders working for seed companies) and in the private sector (university and USDA scientists working on tomato genetics or tomato breeding). An additional target audience reached were scientists at government or university laboratories reached through seminars and outreach to researchers who could use the new tomato lines developed to augment their current research in entomology, virology, or plant moclular biology. Another class of target audience reached this period were undergraduate and graduate students who toured the research field, which were used to teach tomato breeding and integration of disciplines (in this case, genetics, plant pathology, entomology) to solve complex problems Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?An undergraduate student intern was mentored in a research project using the acylsugar lines during the fall 2014/spring 2015 research program. The purpose of the intership was both to provide the undergraduate student with a hands-on research experience, and to train the graduate student acting as the mentor in how to be an effective mentor. The intern presented her results in a cyber symposium in May 2015. The program was overviewed and plants shown to first year Cornell graduate students and senior undergraduates in a lab course fall 2014. The graduate student developing the new acylsugar lines for this project presented his results at the international meeting of the Tomato Breeders Roundtable Fall of 2014, gaining experience in development and presentation of seminars. How have the results been disseminated to communities of interest?The progress of the program was reviewed and plants shown to international seed company breeders and representatives at VBI Field Day, Aug 2015. The PI (Mutschler) also gave a series of semiars on the acylsugar breeding program at universities, goverment laboratories, and seed company headquarters in Hong Kong, Shanghai, Taipei, and Bangkok spring 2015. The PI (Mutschler) also gave a seminar on this topic at the The Xth International Symposium on Thysanoptera and Tospoviruses (Xth ISTT), May 15 - 20, 2015. What do you plan to do during the next reporting period to accomplish the goals?Goal 1: Complete production of lines with combinations of added QTL: FA7/FA8/AS. Characterize the acylsugar production and types of all of the lines in greenhouse trials. Produce seed to generate adequate supplies of seed of all of the lines. Finalize characterization of lines with three epistatic QTL needed for changing sugar of the ayclsugar from sucrose to glucose. Produce seed to establish this line. Although this was not an original goal, if time permits, we will start combining QTL for changing fatty acid type with the 3 QTL needed for the production of acylglucoses. Goal 2. Use all of the new acylsugar lines and controls in regional trials to determine impact of acylsugar chemotype on acylsugar level, type, insect infestation, and virus spread. We have also added in lab/greenhouse tests of the new acysugar lines against a series of important pests, accomplishing some work in the PI and co-PIs lab, and other work with new cooperators who are not on the grant, but are eager to test the lines on the insects that are the focus of their labs. This was not part of the original grant, but is an effort that will demonstrate the potential use of the lines as a platform for working on multiple important pest species. Goal 3. Continue testing of different types of markers/gnotyping (CAPS, SNPS, and genotype by sequencing) to delineate the optimal markers with the different methods, to determine the markers that should be recommended. In the process of goal 1 and goal 3 the data will also allow us to make the gene(s) underlying the acylsugar QTL more finely, and reduce the size of the introgressions carrying these genes. That in turn will refine the markers needed for identification of plants containing the QTL. Goal 4. Proceed with discussion of scope of material to include in webinar, production of videoclips to include in webinar as well as accessory documents to include (such as protocols used in assessing acylsugar level and type).
Impacts
What was accomplished under these goals?
Goal 1.In the prior year's report described the the creation and characterization of the three lines are named FA2/AS, FA7/AS and FA8/AS. For each of these lines, the name indicating the QTL added (FA2 being a chromosome 2 QTL named FA2, etc). In the current year, the additional lines produced are lines combining two FA QTL: FA2/FA7/AS, FA2/FA8/AS, as well as the triple QTL line FA2/FA7/FA8/AS. In greenhouse trials, each of these three lines shows the alteration in the acylsugar fatty acid profile predicted for the QTL added to the line.Also created this year is the line AG3/AG4/AG11, with three QTL that allow it to produce acylglucoses rather than acylsucroses. Initial assays show that this line does produce acylsugars that are >85% acylglucoses. Goal 2. Trials using the new acylsugar lines against whiteflies (FL) and thrips (NC) were performed. FLORIDA (co-PI Smith): Carried out field trials in the fall of 2014 and spring of 2015 at the University of Florida's Gulf Coast Research and Education Center near Tampa to evaluate efficacy of tomato lines from the Cornell breeding program in suppressing sweetpotato whitefly (Bemisia tabaci) and incidence of Tomato yellow leaf curl virus. In the fall of 2014 we tested 15 entries and in the spring of 2015 we tested 11 entries.Data collected on densities of whitefly eggs, first, mid and fourth instar nymphs, and in 2014, data on fruit set. Foliage samples collected for analysis of acylsugar composition.Foliage alsocollected to test for the presence of TYLCV using PCR.Data have beensummarized and analyzed on whitefly numbers and TYLCV incidence associated with each tomato line. NORTH CAROLINA (co-PI Kennedy): Conducted field trial on commercial tomato farm - to evaluate efficacy of tomato lines from the Cornell breeding program in suppressing tobacco thrips and western flower thrips populations and incidence of TSWV. Collected thrips samples for identification to species and performed identifications. Collected foliage samples for analysis of acylsugar composition. Collected and tested foliage for infection by TSWV using ELISA. Summarized and analyzed data on thrips numbers and TSWV incidence associated with each tomato line. Goal 3. The lines were analyzed using CAP markers and also using genotyping by sequencing, to compare the potential of these methods for developing the best means of genotyping lines for presence of the introgressions carrying the targeted QTL Goal 4: some video was made for development of webinars, to test means of developing final materials, and to determine best content.
Publications
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Progress 09/01/13 to 08/31/14
Outputs
Target Audience: The main target audiences reached during this period include tomato breeders in both the private sector (seed company directors of research and tomato breeders working for seed companies) and in the private sector (university and USDA scientists working on tomato genetics or tomato breeding, or on entomology/plant pathology of tomato). An additional target audience were undergraduate students who toured the research field, which were used to teach tomato breeding and integration of disciplines (in this case, genetics, plant pathology, entomology) to solve complex problems. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest? The creation and characterization of the acylsugar sister lines (FA2/AS, FA7/AS, and FA8/AS) were presented by the graduate student on this project at the national plant breeding meeting, Minneapolis MN, Aug 2014. (Poster included acknowledgment of funding). The results of the project were also presented to 40 seed company representatives at the annual field day for the Vegetable Breeding Institute, Aug 2014, Ithaca NY. (Slide presentation included acknowledgment of source of funding). This presentation resulted great interest among seed company representatives. The results of the project were also presented by the graduate student on this project at the international Tomato Breeders Roundtable meeting, Asheville, NC, Sept 2014. (Slide presentation included acknowledgment of source of funding). Attendees (ca. 80) included university faulty and graduate students, governmental scientists, and seed company breeders and research directors. Both presentations at the annual field day for the Vegetable Breeding Institute and the presentation at the Tomato Breeders Roundtable resulted in offers of assistance to expand testing materials in development at numerous sites in the US and abroad, and indications of interest in use of the materials in development for research in control of a wide range of current and emerging insect pests, as well as virus control in tomato . What do you plan to do during the next reporting period to accomplish the goals? Goal 1: a. Complete production of lines with combinations of added QTL: FA2/FA7/AS, FA2/FA8/AS, and FA7/FA8/AS. Characterize the acylsugar production and types in greenhouse trials. Produce seed to establish the lines. b. Continue development of lines with three epistatic QTL needed for changing sugar of the ayclsugar from sucrose to glucose. Characterize the acylsugar production and types in greenhouse trials. Produce seed to establish the lines. Goal 2. Use all of the new acylsugar lines and controls in regional trials to determine impact of acylsugar chemotype on acylsugar level, type, insect infestation, and virus spread. Goal 3. Review new tomato and pennellii sequences, select markers for development of best form of marker analysis for the sister lines, or transfer of QTL. In the process of goal 1 and goal 3 the data will also allow us to make the gene(s) underlying the QTL more finely, and reduce the size of the introgressions carrying these genes. That in turn will refine the markers needed for identification of plants containing the QTL. Goal 4. Proceed with discussion of scope of material to include in webinar, production of video clips to include in webinar as well as accessory documents to include (such as protocols used in assessing acylsugar level and type)
Impacts
What was accomplished under these goals?
GOAL 1. Transfer QTL altering acylsugar chemotype to acylsugar tomato line CU071026, producing sets of sister-lines plus and minus different QTL, and them characterize these lines for acylsugar chemotype and level. The three sister lines whose development was completed this year are FA2/AS, FA7/AS and FA8/AS, with the name indicating the QTL added (FA2 being a chromosome 2 QTL named FA2, etc). In greenhouse trials, each of the three lines shows the alteration in the acylsugar fatty acid profile that were those predicted for the QTL added to the line. This is a very gratifying result Additional lines are still in development, and might be completed in time to be included for Spring 2015 field trials GOAL 2 Determine the impacts of different acylsugar chemotypes produced by the sister lines against whiteflies (FL) and thrips (NC). The sister lines FA2/AS and FA8/AS were included in 2014 spring/summer trials in NC and FL. The samples have all been collected, including acylsugar samples (for level and for fatty acid profile), insect infestation data, and virus data. Some of these samples are still in analysis, which should be completed, and data analyzed by end of 2014. The sister lines FA2/AS, FA8/AS, and FA7/AS are all included in 2014 fall trial that is just starting in FL. GOAL 3. Use genotypic data generated to develop/test acylsugar breeding SNP sets for accelerated introgression of QTL for acylsugar production and controlling acylsugar chemotype into tomato cultivars. We have just staring this work. The substantially updated sequence for cultivated tomato (build 2.5) and the first sequence for S. pennellii (LA716) were both just released summer of 2014. This development greatly impacts the accuracy with which we will be able to address this goal. We have downloaded all of this information, and are working with it and with the SNP data on the benchmark acylsugar line CU071026 to slightly revise/improve our approach to this goal. GOAL 4. Develop content (webinars) describing this project, tomato lines available and teaching methods to breed tomatoes with optimal acylsugar chemotypes for dissemination to breeders via PBGCoP (eXtension). This part was not addressed in the first year.
Publications
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